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Fowler S Zoo and Wild Animal Medicine Current Therapy Volume 9 PDF
Fowler S Zoo and Wild Animal Medicine Current Therapy Volume 9 PDF
FOWLER’S
ZOO and
WILD ANIMAL
MEDICINE
Current Therapy
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VOLUME 9
FOWLER’S
ZOO and
WILD ANIMAL
MEDICINE
Current Therapy
R. Eric Miller, DVM, DACZM
Executive Director, WildCare Institute
Saint Louis Zoo
St. Louis, Missouri
Adjunct Associate Professor of Veterinary Medicine and Surgery
College of Veterinary Medicine
University of Missouri
Columbia, Missouri
Previous editions copyrighted 2015, 2012, 2008, 2003, 1999, 1993, 1986, 1978 by Saunders, an imprint of
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Printed in China
This is a sad introduction to write • The AAZV’s Murray Fowler Scholarship Fund supports
but also one that is celebratory. It the attendance of international zoo and wildlife col-
is sad because Murray Fowler died leagues at the AAZV’s annual meeting
in May 2014 and this is the first • * The European Association of Zoo and Wildlife Veteri-
of nine editions of Zoo and Wild narians (EAZWV) Honorary Membership and
Animal Medicine, the book he initi- • The Iowa State Alumnus Award.
ated, without Murray as an editor. Yet, Murray was much more than just an author or
This introduction is also celebratory an editor, or a member of any single group. He was a
because it summarizes a life lived to veterinarian who very much belonged to our worldwide
the fullest with so many around the zoo community. The most recent edition of Zoo and Wild
world and lived to the fullest with Animal Medicine (ZAWAM)—the 8th, which he received
Audrey and his family. Nearly everyone in the zoo and just before his death, illustrated that world view. Its 82
wildlife community has vivid, caring memories of Murray chapters include authors from 17 countries on 6 continents.
Fowler, so I will humbly try to provide some thoughts in a In another example, when the Iron Curtain separated so
relatively brief tribute. many people in Europe, he and Rudolph Ippen from East
Murray’s resume is legendary: he was a founding member Germany worked beyond political boundaries to create an
of the American College of Veterinary Internal Medicine international alliance for our profession. I am comforted
(ACVIM), the American College of Veterinary Toxicology believing that somewhere Murray and Rudolph are sharing
(ACVT), and the American College of Zoological Medicine good memories and celebrating the future of our field they
(ACZM). He was also a member of the American Associa- did so much to establish.
tion of Zoo Veterinarians (AAZV), a President of nearly On a personal level, it was also clear to me how deeply
every organization in our field, and a member of even more, Murray cared for individual zoo and wildlife veterinarians
including the European Association of Zoo and Wildlife around the world when he and Audrey and my wife and
Veterinarians (EAZWV) and the Brazilian zoo veterinarians I were able to attend meetings and symposiums together.
(they even call it Groupo Fowler). I heard many times how Murray and Audrey had been
Murray wrote or edited more than 25 books—on zoo welcomed in so many places, and how those experiences led
and wildlife medicine, New World camelids, elephants, to lifelong friendships that meant so much to them.
toxicology, and wild animal restraint. All were always A typical sentiment about Murray came from Teresa
hands-on and practical texts! And I would note that Fernandes, a veterinarian at the Lisbon Zoo: At her first
Audrey, his wife, was a behind-the-scenes hero of many of AAZV meeting in Tulsa, she and a friend from Moscow met
these books—she was the quiet editor who ensured that Murray—she wrote, “He was kind enough to exchange a
the books always included proper grammar and sentence few nice words with us and shake hands with us, complete
structure! strangers to him. We had come all the way from Europe
Murray’s achievements are many. For example, in 1976 and yet this moment made it all worth it by itself. Never in
Murray founded the first residency in zoo and wildlife my wildest dreams did I think I would be having a private
medicine at the UC-Davis. He contributed much to zoo dinner with the ‘father’ of zoological medicine.” I believe
and wildlife medicine during sabbaticals in Germany, that she, like so many zoo and wildlife veterinarians around
Denmark, San Diego, the United Kingdom, and Uganda. the world, were part of Murray’s “zoo family,” a family that
In recognition of these achievements and many others, he he deeply treasured.
received numerous awards. The following are but a few I would like to offer one more illustration of his caring
examples: nature: Working with him on the 4th edition of ZAWAM
• The American Association of Zoo Veterinarian’s (AAZV) was the first time I had edited a textbook. I had never been
Dolensek Award asked for autographs, but when the requests came in, I
• The Association of Zoos and Aquariums (AZA) Marlin signed “Best wishes, Eric”—that was until I saw Murray’s
Perkins Award autographs. He wrote deep, touching and personal notes
• The British Zoological Veterinary Society’s (BZVS) Park because he had taken the time to talk to each recipient
Davis Award and to hear each person’s own history and hopes. That
v
vi A Tribute to Murray E. Fowler
experience resulted in our only negative interaction, as I In conclusion, when asked how he would like to be
scolded Murray for not offering me better training! Those remembered, Murray once said, “I guess I would like to be
autographs also illustrated how much he loved teaching and remembered as a capable veterinarian, with an interest in a
how much he loved students. In interactions, both brief and broad range of different species of animals, and as a teacher
years long, Murray always encouraged everyone that he met with a desire to share in the written and oral form as much
to be their best and to pursue their dreams. As I wrote this information that I have.” That was an understatement.
memoriam, another became clear to me—frankly, if you The word giant is overused. And despite his previous
were not a friend of Murray’s, you simply had not had a modest statement, Murray truly was a giant—not only in
chance to meet him. veterinary medicine, but in life. I believe that the best way
I will repeat the acknowledgement I wrote for the 7th we can honor Murray is to continue developing our profes-
edition of ZAWAM: “Dr. Fowler initiated the first edition sion of zoo and wildlife medicine in the way I believe he
of Zoo and Wild Animal Medicine in 1978, when few texts would have wanted. That will require not only committing
existed in the field of zoo and wildlife medicine. In the our best efforts to caring for the animals entrusted to us,
subsequent 32 years, he has shown an unwavering dedica- but also by being kind to each other and above all, teaching,
tion to the dissemination of this information with seven mentoring, and sharing with both students and our peers
subsequent volumes of this text—not to mention many in our field. That is the way that Murray will live on in
other related books authored by him. He has been, and each of us. I owe a personal thank you, and I believe we
continues to be, a mentor and an inspiration to many in our all owe a collective thank you, to Murray for everything
field, myself included.” When he edited that acknowledge- that he did for our profession and for us individually.
ment, in typical Murray fashion, his comment was, “Well, His memory will live on in the title of this book and in
that is a bit over the top.” our hearts.
Contributors
Michael J. Adkesson, DVM, DACZM, DECZM (ZHM) Cheryl Asa, BA, MS, PhD
Vice President, Clinical Medicine Affiliate Scientist
Chicago Zoological Society Saint Louis Zoo
Brookfield Zoo St. Louis, Missouri;
Brookfield, Illinois; Advisory Board Chair
Adjunct Clinical Assistant Professor AZA Reproductive Management Center
Department of Veterinary Clinical Medicine Saint Louis Zoo
College of Veterinary Medicine, University of Illinois St. Louis, Missouri
Urbana, Illinois Changes in Reproductive Management
Use of Computed Tomography/Magnetic Resonance Imaging
in Zoological Medicine Kay A. Backues, DVM, DACZM
Systemic Isosporosis in Passerine Birds Director of Animal Health
Veterinary Health Department
Patricia Aguilar-Calvo Tulsa Zoo
Postdoctoral Fellow Tulsa, Oklahoma;
Pathology Adjunct Professor
University of California San Diego Lab Animal and Exotic Pet Medicine
San Diego, California Tulsa Community College
Prion Diseases in Wildlife Tulsa, Oklahoma;
Adjunct Professor
Bianca Nascimento de Alcantara, MSc Zoo-Exotic Medicine Service
PhD Candidate Oklahoma State University
Arboviology Section and Hemorrhagic Fevers Stillwater, Oklahoma
Pathology Section Elephant Mycobacteriosis: New Diagnostics and Management
Evandro Chagas Institute
Ananindeua, Pará, Brazil James E. Bailey, DVM, MS, DACVAA
Zika Virus: A Real Threat to Wildlife? President
Anesthesiology
Matthew C. Allender, DVM, MS, PhD, DACZM Innovative Veterinary Medicine, Inc.
Director Ponte Vedra, Florida
Wildlife Epidemiology Lab Lens Diseases and Anesthetic Considerations for
University of Illinois Ophthalmologic Procedures in Pinnipeds
Urbana, Illinois;
Assistant Professor Karen Bauman, MS
Department of Veterinary Clinical Medicine Laboratory Manager
University of Illinois Research Department
Urbana, Illinois Saint Louis Zoo
Ranaviral Disease in Reptiles and Amphibians St Louis, Missouri
Wildlife Technologies
Leonardo Arias-Bernal, DVM, MSc
Director
Bioparque WAKATA
Parque Jaime Duque
Bogotá, Austria;
Professor
Veterinary Medicine Program
Universidad de La Salle
Bogotá, Bogotá
Medicine of Captive Andean Bears
vii
viii Contributors
Carmen M.H. Colitz, DVM, PhD Marion Renée Desmarchelier, DVM, IPSAV, DES, MSc,
All Animal Eye Care, Inc. DACZM
Jupiter, Florida Clinical Instructor
Lens Diseases and Anesthetic Considerations for Zoological Medicine Service
Ophthalmologic Procedures in Pinnipeds Faculté de médecine vétérinaire - Université de Montréal
Saint-Hyacinthe, QC, Canada
Galaxia Cortes-Hinojosa, MV, MSc, PhD A Systematic Approach in Diagnosing Behavior Problems
Postdoctoral Associate
Biology Nicola Di Girolamo, DMV, MSc(EBHC), PhD, DECZM
University of Florida (Herpetology)
Gainesville, Florida Tai Wai Small Animal & Exotic Hospital, Hong Kong
Marine Mammal Viruses EBMVet, http://ebmvet
Doctor
José Luis Crespo-Picazo, LV Clinica per Animali Esotici
Head of Conservation Programs Veterinary Specialists Center
Veterinary Services & Research Department Roma, Italy;
Fundación Oceanogràfic, Avanqua Oceanogràfic-Ágora Veterinary Sciences
Valencia, Spain University of Bologna
Decompression Medicine in Aquatic Species (Fish and Sea Ozzano Emilia, BO, Italy
Turtle Focus) Research Study Design
Brett Fundak, BS, MS, DVM, DACVR Martin Gilbert, BVMS, PhD, MRes, BSc (Hons)
Staff Radiologist Senior Research Associate
Antech Imaging Services Department of Population Medicine and Diagnostic
Private Corporation Sciences
Fountain Valley, California Cornell University, College of Veterinary Medicine
Moving Beyond Survey Radiographs Ithaca, New York
Techniques for Vaccinating Wildlife
Laurie J. Gage, DVM, DACZM
Big Cat and Marine Mammal Specialist Steven M. Goodman, PhD, HAB
USDA APHIS Animal Care MacArthur Field Biologist
Center for Animal Welfare Field Museum of Natural History
Kansas City, Missouri Chicago, Illinois;
Giraffe Husbandry and Welfare Scientific Counselor
Association Vahatra
Kathryn C. Gamble, DVM, MS, DACZM, Antananarivo, Madagascar
DECZM (ZHM) Disease Risk to Endemic Animals from Introduced Species
Dr. Lester E. Fisher Director of Veterinary Medicine on Madagascar
Veterinary Services
Lincoln Park Zoo Mark Greenberg, MD
Chicago, Illinois Professor of Anesthesiology and Pediatrics
Compounding Pharmacies Anesthesiology and Pediatrics
Antifungals in Birds University of California-San Diego
San Diego, California;
Daniel García-Párraga, LVet, DECZM (ZHM), DECAAH Director of Pediatric Anesthesiology and Critical Care
Director of Animal Health Medicine
Oceanografic Anesthesiology
Valencia, Spain; University of California-San Diego
Scientific Director San Diego, California
Oceanografic Foundation Use of Naltrexone and Atipamezole in Emergency Response
Valencia, Spain to Human Exposure to Ultra-Potent Opioids and Alpha-2
Decompression Medicine in Aquatic Species (Fish and Sea Agonists in Zoo and Wildlife Medicine
Turtle Focus)
Alex David Greenwood, BA, PhD
Michael M. Garner, DVM, DACVP Department Head
Founder and Director Wildlife Diseases
Northwest ZooPath Leibniz Institute for Zoo and Wildlife Research
Monroe, Washington Berlin, Germany;
Avian Spirurids Professor
Veterinary Medicine
Timothy A. Georoff, VMD, DACZM Freie Universität Berlin
Associate Veterinarian Berlin, Germany
Philadelphia Zoo Equine Herpesviruses and Interspecies Infections
Philadelphia, Pennsylvania
Canine Distemper Vaccination in Nondomestic Carnivores Carsten Grøndahl, DVM, PhD
Chief Veterinarian
Kirsten V.K. Gilardi, DVM Veterinarian Department
Karen C. Drayer Wildlife Health Center Copenhagen Zoo
School of Veterinary Medicine Frederiksberg, Germany
University of California, Davis Musk Ox Sedation and Anesthesia
Davis, California
The USAID Emerging Pandemic Threats PREDICT Catherine Hadfield, MA, VetMB, DACZM, DECZM
Project—Global Detection of Emerging Wildlife Viral Senior Veterinarian
Zoonoses Seattle Aquarium
Seattle, Washington
Disease Risks to Native Wildlife From Zoos and Aquariums
Touch-Pools: The Other Side of the Hand
Contributors xi
Kerrie Anne T. Loyd, PhD Jonna A.K. Mazet, DVM, MPVM, PhD
Lecturer Professor & Executive Director
Biology One Health Institute
Arizona State University UC Davis School of Veterinary Medicine
Lake Havasu City, Arizona Davis, California
Feral Cat Dilemma The USAID Emerging Pandemic Threats PREDICT
Project—Global Detection of Emerging Wildlife Viral
Zoonoses
Contributors xiii
Francisco Olea-Popelka, DVM, MSc, PhD Timothy J. Portas, BVSc, MVSc, MACVS, DACZM
Assistant Professor Zoo and Wildlife Veterinary Consultancy
Clinical Sciences North Maleny
College of Veterinary Medicine and Biomedical Sciences, Queensland, Australia
Colorado State University Medical Aspects of Potoroid Marsupial Conservation
Fort Collins, Colorado Translocations
A Practical Guide for Statistics in Wildlife Studies
Robin W. Radcliffe, DVM, DACZM
Klaus Osterrieder, Dr. Med. Vet., Senior Lecturer
Dr. Med. Vet. Habil. Department of Clinical Sciences
Professor of Virology and Chair Cornell University
Institut für Virologie Ithaca, New York;
Freie Universität Berlin Head, Cornell Conservation Medicine Program
Germany Cornell University
Equine Herpesviruses and Interspecies Infections Ithaca, New York;
Faculty Fellow
Annie Page-Karjian, DVM, PhD Atkinson Center for a Sustainable Future
Assistant Research Professor & Clinical Veterinarian Cornell University
Harbor Branch Oceanographic Institute Ithaca, New York
Florida Atlantic University Health of the Forest Rhinoceroses of Southeast Asia:
Fort Pierce, Florida; Sumatran and Javan Rhinoceroses
Affiliate Research Professor
Department of Biological Sciences Jan Raines, DVM
Florida Atlantic University Veterinarian
Boca Raton, Florida Dallas Zoo
Fibropapillomatosis in Marine Turtles Dallas, Texas
Naked Mole Rat Management and Medicine
Jean A. Paré, DMV, DVSc, DACZM
Senior Veterinarian Bonnie L. Raphael, DVM, DACZM
Zoological Health Program WCS Veterinary Consultant
Wildlife Conservation Society Zoological and Wildlife Veterinary Consulting and
Bronx, New York Services
Ophidiomycosis New York, New York
Rehabilitation Medicine of Confiscated Turtles
Adriana Pastor, BSc, DVM
Assistant Director of Veterinary Care Fidisoa Rasambainarivo, DVM, MSc
Animal Health Department of Biology
San Antonio Zoo University of Missouri St Louis
San Antonio, Texas St. Louis, Missouri
Babesiosis in Cervidae Madagascar Fauna and Flora Group
Toamasina, Madagascar
Joost Philippa, DVM, PhD Disease Risk to Endemic Animals from Introduced Species
Wildlife Veterinarian on Madagascar
Kigali, Rwanda
Infectious Diseases of Orangutans in Their Home Ranges Patricia Reed, DVM
and in Zoos Field Veterinarian
Cape Town, South Africa
Wouter Pieters, DVM Ebola Virus Disease in Great Apes
Veterinarian
Oasis Park Fuerteventura Jack C. Rhyan, DVM, MS
La Lajita Veterinary Medical Officer
Las Palmas, Spain National Wildlife Research Center
Capripoxviruses in Nondomestic Hoofstock Fort Collins, Colorado
Brucellosis in North American Wildlife
Contributors xv
Bruce Rideout, DVM, PhD David Sanchez-Migallon Guzman, LV, MS, DECZM
Director, Disease Investigations (Avian), DACZM
Institute for Conservation Research Veterinary Teaching Hospital
San Diego Zoo Global UC Davis School of Veterinary Medicine
San Diego, California Davis, California
Disease Risks to Native Wildlife from Zoos and Aquariums Sustained-Release and Long-Acting Opioid Formulations
of Interest in Zoological Medicine
John Roberts, BEng, Materials Science & Engineering
Director of Elephants & Conservation Willem Schaftenaar, DVM
Golden Triangle Asian Elephant Foundation Veterinary Advisor European Elephant TAG
Group Director of Sustainability and Conservation Veterinary Department
Minor Hotel Group Rotterdam Zoo
Chiang Saen, Chiang Rai, Thailand Rotterdam, the Netherlands
Elephant Care in Southeast Asia Elephant Endotheliotropic Herpesvirus
Elizabeth Marie Rush, BS, DVM, DACZM Christina J. Sigurdson, DVM, PhD
Staff Specialist Associate Professor
Imaging-Zoo, Wildlife, Exotics Pathology
Antech Imaging Services University of California, San Diego
Irvine, Alabama; La Jolla, California;
Associate Professor, Coordinator Zoo and Wildlife Associate Professor
Research Pathology, Microbiology, Immunology
Pathobiology University of California, Davis
St. George’s University School of Veterinary Medicine Davis, California
True Blue, Indonesia Prion Diseases in Wildlife
Moving Beyond Survey Radiographs
xvi Contributors
Kurt K. Sladky, MS, DVM, DACZM John M. Sykes IV, DVM, DACZM
Professor, Zoological Medicine Clinical Department Head, Zoological Health Program
Surgical Sciences Marilyn M. Simpson Distinguished Veterinarian
University of Wisconsin School of Veterinary Medicine Wildlife Conservation Society
Madison, Wisconsin Bronx, New York
Reptile and Amphibian Analgesia Opportunities to Inspire the Next Generation of Veterinarians
Dale Smith, DVM, DVSc Jessica J. Talbot, BSc (Vet.) (Hons), BVSc (Hons)
Professor Doctor
Department of Pathobiology The University of Sydney
Ontario Veterinary College, University of Guelph Camperdown, New South Wales, Australia
Guelph Quality-of-Life Assessment and End-of-Life Planning for
Ontario, Canada Geriatric Zoo Animals
Bornaviruses in Birds
Washington Tapia, MSc
Kristine Smith, DVM, DACZM Director of Giant Tortoise Restoration Initiative
EcoHealth Alliance Galapagos Conservancy, Inc.
Health and Policy Puerto Ayora, Galápagos, Ecuador
Trumbull, Connecticut Medical Aspects of Giant Tortoise Relocation in the
Risk Analysis Framework Guidance for Wildlife Health Galápagos Islands
Professionals
Karen A. Terio, DVM, PhD, DACVP
Endre Sós, DVM, PhD, DECZM (ZHM) Clinical Associate Professor
Head Veterinarian, Director Zoological Pathology Program
Directorate of Conservation and Veterinary Services University of Illinois
Budapest Zoo Brookfield, Illinois
Budapest, Hungary Systemic Isosporosis in Passerine Birds
Medical Aspects of the Hungarian Meadow Viper
Reintroduction Scott Terrell, DVM, DACVP
Director, Animal and Science Operations
Gerhard Steenkamp, BSc, BVSc, MSc Disney’s Animals, Science, and Environment
Senior Lecturer Disney Parks and Resorts
Companion Animal Clinical Studies Orlando, Florida
Faculty of Veterinary Science, University of Pretoria Strategic Planning for Zoo Veterinary Operations
Pretoria
Gauteng, South Africa Arshad Haroon Toosy, DVM, MSc (Hons)
Management of Dental Disease in Elephants Manager Veterinary Services
Life Science
Darrel K. Styles, DVM, PhD Al Ain Zoo
USDA APHIS Veterinary Services Al Ain
Riverdale, Maryland Abu Dhabi, United Arab Emirates
Avian Influenza: A Brief Overview of the Pathobiology and An Overview of Middle East Respiratory Syndrome
Current Status in Domestic and Nondomestic Species in the Middle East
Kathryn A. Tuxbury, MS, DVM Chris Walzer, Univ. Prof. Dr. Med. Vet., DECZM
Associate Veterinarian (Wildlife Health)
Animal Health Department Chair Conservation Medicine
New England Aquarium Department of Integrative Biology and Evolution
Boston, Massachusetts University of Veterinary Medicine Vienna
Touch-Pools: The Other Side of the Hand Vienna, Austria;
Exec. Director Wildlife Health Program
Eduardo V. Valdes, BSc (Agr.), MSc, PhD Wildlife Conservation Society
Operation Manager-Animal Nutrition Center Bronx, New York
Animal Health International Sample Movement: Overview of Convention
Walt Disney World on International Trade in Endangered Species of Wild
Lake Buena Vista, Florida; Fauna and Flora and Selected National Regulations
Adjunct Professor
Biology Jim Wellehan, DVM, MS, PhD, DACZM, DACVM
University of Central Florida (Virology, Bacteriology/Mycology)
Orlando, Florida; Associate Professor
Adjunct Professor Zoological Medicine Service
Animal Sciences University of Florida College of Veterinary Medicine
University of Florida Gainesville, Florida
Gainesville, Florida Marine Mammal Viruses
Update on Rhinoceros Nutrition
Ellen Wiedner, VMD, DACVIM (Large Animal)
Caroline Van Hemert, PhD Clinical Lecturer in Zoo & Wildlife Medicine
Research Wildlife Biologist University of Florida
US Geological Survey Alaska Science Center College of Veterinary Medicine
Anchorage, Alaska Gainesville, Florida
The Effects of Climate Change on Disease Spread in Wildlife Elephant Mycobacteriosis: New Diagnostics and Management
In many ways, this is a sad edition of Zoo and Wild Animal veterinarians (e.g., leadership, occupational health, educa-
Medicine (ZAWAM) to edit as it is the first without Murray tion of the public [children] about our profession, sample
Fowler. He was truly one of the giants of the field of zoo and importation, the role of animal welfare, and decision
wildlife medicine (see tribute in this edition). Appropriately, making, as well as One Health, and emerging diseases).
it will always be called “Fowler’s” ZAWAM. At the same The challenges for zoo and wildlife medicine, and for the
time, it is a pleasure to welcome two new co-editors, Paul veterinarians who implement it, are worldwide. Therefore,
Calle and Nadine Lamberski. as in previous editions, the authors are an international
This 9th edition contains 100 chapters and returns to the group and in this edition represent 115+ authors from 21
current veterinary therapy format featuring focused, special- countries on 6 continents: Argentina, Austria, Australia,
ized topics, which will also be the focus of the 10th edition. Brazil, Canada, Colombia, Denmark, France, Hungary,
The last (8th) edition featured the taxa-based approach, Germany, Hong Kong, Indonesia, Madagascar, Mexico,
which will also be the focus of the 11th edition. Peru, South Africa, Spain, The Netherlands, United Arab
This volume contains many taxa-based clinical topics, Emirates, the United Kingdom, and the United States.
but also additional issues important to zoo and wildlife
xviii
Acknowledgments
As with previous editions, the authors freely shared their information and time for
the benefit of wild animals and the people who care for them. Therefore, our special
thanks goes to the authors who contributed to this edition of Fowler’s Zoo and Wild
Animal Medicine, as all of the royalties go to support wild animal health research (the
Morris Animal Foundation Wildlife Health Fund and the Wild Animal Health Fund
of the American Association of Zoo Veterinarians) with no financial support going to
the authors or editors. Animal health is a team effort, and we would also like to thank
the animal health and care teams that work with us to maintain animal health and welfare.
We also extend our deep and heartfelt thanks to our family and friends who supported
us through our editorial endeavors.
Last, but certainly not least, we thank the following consulting editors who submitted
potential topics for this edition:
Consulting Editors
Kathryn C. Gamble, DVM, MS, DACZM, DECZM (ZHM)
Lincoln Park Zoo
Chicago, Illinois
xix
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Contents
6. Opportunities to Inspire the Next Generation 17. Disease Risks to Native Wildlife From Zoos
of Veterinarians, 28 and Aquariums, 99
John M. Sykes IV Bruce Rideout and Catherine Hadfield
7. Strategic Planning for Zoo Veterinary 18. Feral Cat Dilemma, 104
Operations, 34 Kerrie Anne T. Loyd and Sonia Maria Hernandez
Scott Terrell
19. The United States Agency for International
8. Organizational Influence: Navigating Development Emerging Pandemic Threats
the Leadership Road for Zoo PREDICT Project—Global Detection of
Veterinarians, 39 Emerging Wildlife Viral Zoonoses, 110
Donald L. Janssen Kirsten V.K. Gilardi and Jonna A.K. Mazet
9. Contingency Planning for All Hazards and 20. Renewable Energy: Effects on Wildlife, 117
Foreign Animal Disease, 45 Peregrine L. Wolff and Michael R. Schirmacher
Yvonne Nadler
Section 4: Reproduction
10. Veterinary Occupational Health and Safety in
the Zoo and Wildlife Setting, 53 21. Female Infertility in Zoo Animals, 124
Elizabeth E. Hammond Bruce Christensen
xxi
xxii Contents
23. Issues Surrounding Surplus Animals 35. Chagas Disease: Wildlife Infection With
in Zoos, 134 Trypanosoma Cruzi in a One Health
Mads Frost Bertelsen Context, 239
Sarah Hamer and Carolyn Hodo
Section 5: Therapeutics
36. The Effects of Climate Change on Disease
24. Stem Cell Therapy in Zoo Medicine, 138 Spread in Wildlife, 247
Matthew E. Kinney and Robert Harman Erik Hofmeister and Caroline Van Hemert
Section 6: Anesthesia and Analgesia 38. Avian Influenza: A Brief Overview of the
Pathobiology and Current Status in Domestic
26. Sustained-Release and Long-Acting Opioid and Nondomestic Species, 262
Formulations of Interest in Zoological Darrel K. Styles and Yvonne Nadler
Medicine, 151
Jessica A. Emerson and David Sanchez-Migallon Guzman 39. Emerging Reptile Viruses, 267
Rachel E. Marschang
27. Use of Naltrexone and Atipamezole
in Emergency Response to Human 40. Emerging Diseases in Bats, 274
Exposure to Ultra-Potent Opioids and Jonathan H. Epstein
Alpha-2 Agonists in Zoo and Wildlife
Medicine, 164 41. Zika Virus: A Real Threat to Wildlife?, 280
Jeffery R. Zuba and Mark Greenberg Lilian Silva Catenacci and Bianca Nascimento de Alcantara
28. Vaporizers and Field Anesthesia Equipment 42. An Overview of Middle East Respiratory
for Free-Ranging Wildlife, 177 Syndrome in the Middle East, 287
Sathya K. Chinnadurai Arshad Haroon Toosy and Sean O’Sullivan
29. Perianesthetic Monitoring: Equipment and 43. Disease Risk to Endemic Animals From
Interpretation, 185 Introduced Species on Madagascar, 292
Khursheed Mama Fidisoa Rasambainarivo and Steven M. Goodman
31. Use of Computed Tomography/ Magnetic 45. Brucellosis in North American Wildlife, 306
Resonance Imaging in Zoological Jack C. Rhyan and Pauline Nol
Medicine, 206
Michael J. Adkesson and Marina Ivančić 46. Update on Melioidosis in Zoo and Wild
Animals, 315
32. Moving Beyond Survey Radiographs, 218 Paolo R. Martelli and Hui Suk-Wai
Elizabeth Marie Rush and Brett Fundak
Section 10: Aquatic
Section 8: Emerging and Changing 47. Techniques for Addressing Parasites in
Infectious Diseases Saltwater Aquariums, 323
Claire Erlacher-Reid
33. Equine Herpesviruses and Interspecies
Infections, 227 48. Touch-Pools: The Other Side of the Hand, 334
Alex David Greenwood and Klaus Osterrieder Catherine Hadfield and Kathryn A. Tuxbury
34. Ebola Virus Disease in Great Apes, 233 49. Sharks and Medicine, 338
Kenneth Cameron and Patricia Reed Leigh Clayton and Kathryn E. Seeley
Contents xxiii
52. Ranaviral Disease in Reptiles and 68. Selected Medical Aspects of Bird
Amphibians, 364 Reproduction in Ex Situ Conservation, 481
Matthew C. Allender Dante Luis Di Nucci
Section 16: Great Apes 90. Musk Ox Sedation and Anesthesia, 636
Carsten Grøndahl
80. Infectious Diseases of Orangutans in their
Home Ranges and in Zoos, 565 91. Capripoxviruses in Nondomestic
Joost Philippa and Rosalie Dench Hoofstock, 641
Wouter Pieters
81. Medical Aspects of Chimpanzee
Rehabilitation and Sanctuary 92. Babesiosis in Cervidae, 647
Medicine, 574 Adriana Pastor and Ellie Milnes
Jocelyn Bezner
Section 19: Elephants and Rhinoceroses
82. Update on the Great Ape Heart
Project, 581 93. Management of Dental Disease in
Hayley Weston Murphy and Marietta Dindo Danforth Elephants, 657
Gerhard Steenkamp
83. Great Ape Nutrition, 588
Debra A. Schmidt and Michelle E. Shaw 94. Elephant Mycobacteriosis: New Diagnostics
and Management, 665
Section 17: Marine Mammals Kay A. Backues and Ellen Wiedner
84. Marine Mammal Viruses, 597 95. Elephant Endotheliotropic Herpesvirus, 672
Jim Wellehan and Galaxia Cortes-Hinojosa Lauren Lynn Howard and Willem Schaftenaar
85. Mycobacterium pinnipedii, 603 96. Elephant Pregnancy and Parturition: Normal
Alexis Lécu and Abnormal, 680
Imke Lüeders
86. Lens Diseases and Anesthetic Considerations
for Ophthalmologic Procedures in 97. Elephant Care in Southeast Asia, 689
Pinnipeds, 610 Gerardo Martinez and John Roberts
Carmen M.H. Colitz and James E. Bailey
98. Updates in African Rhinoceros Field
Section 18: Ruminants Immobilization and Translocation, 692
Pete Morkel and Pierre Nel
87. Giraffe Husbandry and Welfare, 619
Laurie J. Gage 99. Update on Rhinoceros Nutrition, 699
Kathleen E. Sullivan and Eduardo V. Valdes
88. Lameness Diagnosis and Management in Zoo
Giraffe, 623 100. Health of the Forest Rhinoceros of
Liza Dadone Southeast Asia: Sumatran and Javan
Rhinoceros, 707
89. Mass Mortality Events Affecting Saiga Robin W. Radcliffe and Kurnia Oktavia Khairani
Antelope of Central Asia, 630
Richard Anthony Kock and Sarah Robinson Index, 716
SECTION 1
General
1 The Role of Veterinary Advisors in Animal
Management Plans, 2
3 Wildlife Technologies, 11
1
1
The Role of Veterinary Advisors in
Animal Management Plans
JULIA E. NAPIER
Introduction in 1994 and has grown considerably since then (Table 1.1).
The disparity in numbers from 1994 to 20162 provides a
The Veterinary Advisor is a member of the Veterinary wealth of opportunities for zoo veterinarians to contribute
Advisory Group (VAG), which is a subcommittee of the their knowledge, time, and energy to a population that
Association of Zoos and Aquariums (AZA) Animal Health needs a Veterinary Advisor in the VAG.
Committee (AHC) and was established with three main Currently, the subcommittee is composed of SSP/TAG
goals in mind: Veterinary Advisors (which includes clinicians and patholo-
1. To act as a support and advisory body for Species Survival gists) and in a few instances, a nutritionist. The VAG Chair
Plans (SSP)/Taxon Advisory Groups (TAG) Veterinary is appointed by the Chair of the AZA AHC. The original
Advisors; “Guidelines for Veterinary Advisors to Regional Conserva-
2. To act as a source of information for protocols concern- tion Plans” was submitted to the AZA Wildlife Conserva-
ing the roles and responsibilities of Veterinary Advisors; tion and Management Committee (WCMC) and accepted
and in 1993. These were revised in 1994 and again in 2001
3. To serve as an informational resource on veterinary issues and 2009. Another revision is underway as of the writing
that may impact conservation programs. of this chapter. The benefits of these guidelines are twofold:
(1) they offer the SSP Coordinator a reasonable expecta-
Background tion of the role of a Veterinary Advisor, and (2) they offer
the Veterinary Advisor an outline of basic standards that
The concept of the VAG originated with the Infectious should be met. Clearly, the exact role and responsibilities
Disease Committee (IDC) of the American Association of the Veterinary Advisor will differ among the various SSP/
of Zoo Veterinarians (AAZV) and the Conservation and TAG programs. The list of SSP/TAG Veterinary Advisors
Science Department of AZA, formerly the American is maintained by the VAG Chair and is updated as neces-
Association of Zoological Parks and Aquariums (AAZPA) sary. This list is posted on the AAZV website.3 The AAZV
in 1993. The need for such a group was highlighted at the site was chosen to encourage use by zoo veterinarians. In
1992 International Conference on Implications of Infec- addition to the list of advisors, SSP/TAG veterinary and
tious Diseases for Captive Propagation and Reintroduction necropsy protocols, and Annual Report Forms are posted
Programs of Threatened Species. This meeting, held in as they become available.
Oakland, California, was sponsored by AZA, AAZV, and
the Captive Breeding Specialist Group of the International Veterinary Advisor Responsibilities
Union for the Conservation of Nature Species Survival Com-
mission (CBSG/IUCN/SSC).1 It emphasized the impact The traditional role of the Veterinary Advisor as stated in the
of disease on reintroduction projects and highlighted the guidelines includes making recommendations for diagnostic
importance of risk assessment. A general lack of informa- testing and evaluations, providing appropriate laboratory
tion on (1) incidence, distribution, and risks of disease information for disease testing in their specific taxa, sug-
in captive and wild populations, (2) effective quarantine gesting therapeutic protocols for treatment of disease and
protocols necessary to prevent disease transmission, and (3) anesthetic protocols for immobilizations, and developing
definitive diagnostic tests to detect and monitor disease had successful quarantine and preventative medicine measures.
resulted in the lack of a working database for informed risk However, especially in the past 10–15 years, the role of
assessment. The notion of a Veterinary Advisor to each SSP/ the Veterinary Advisor as well as the clinical veterinarian
TAG program was put forward as a way to generate and in a zoological setting has evolved into something much
collect this missing information. The program was created more complex than just practicing medicine, formulating
2
CHAPTER 1 The Role of Veterinary Advisors in Animal Management Plans 3
TABLE Comparison of Species Survival Plans, Taxon The Veterinary Advisor has the unique ability not just
1.1 Advisory Groups, and Veterinary Advisor to advocate for an animal’s well-being in every aspect, but
Numbers From Inception in 1994–2016 they are an invaluable resource to collection veterinarians, to
specific managed populations in both captive and free-range
Species Taxon settings, and to in situ and ex situ conservation field proj-
Survival Advisory Veterinary % Vets to ects involving their species of expertise. They also benefit
Plans Groups Advisor Programs students, animal care staff, and veterinarians whose primary
1994 69 41 82 75 focus is not exotic species, as well as human medicine experts
who may volunteer their time to the zoo community.
2016 611* 46 132 20
4
CHAPTER 2 Risk Analysis Framework Guidance for Wildlife Health Professionals 5
dissemination, management option evaluation, etc.) is the • Conduct a background literature and data search on the
goal of risk communication. This is an important, but problem—this often includes both scientific and policy
often overlooked, aspect of the risk analysis continuum information.
and should take place throughout the entire process. • Create a list of all potential hazards (usually diseases in
this case) that can then be prioritized through the hazard
Wildlife Disease Risk and the Wildlife Interface identification phase.
Since 1992, the Conservation Breeding Specialist Group • Establish the group’s acceptable level of risk—the level at
(CBSG, now the Conservation Planning Specialist Group which stakeholders will require management action op-
[CPSG]) of the International Union for Conservation of tions. This accepts the basic premise that “zero risk” does
Nature Species Survival Commission (IUCN-SSC) has not occur, or rarely occurs. Zero potential morbidity or
been facilitating collaboration between experts in zoo and likelihood of disease transmission is often not realistic.
wildlife veterinary medicine, disease ecology, and popula- After Problem Formulation, the hazard identification
tion management to develop a set of methods and tools for and then the risk assessment phase (including pathways
realistic and rigorous analysis of disease risks in wildlife, and threats, vulnerability and consequence assessments) are
and at the wildlife–domestic animal–human interface. In undertaken.
2010, recognizing that the range of concerns in relation to For illustrative purposes, this chapter presents one
wildlife disease had broadened well beyond those associated example of adaptive use of this framework in the form of
with animal movements, the OIE and IUCN sponsored the a qualitative wildlife disease risk analysis exercise. A recent
publication of the Manual of Procedures for Wildlife Disease disease risk analysis was undertaken by the University of
Risk Analysis and its companion, the IUCN Guidelines for Minnesota, EcoHealth Alliance, and Food Systems Institute
Wildlife Disease Risk Analysis.5,6 The intent of these publica- in partnership with the US Department of Homeland
tions is to assist in the implementation of risk assessment Security (DHS). The goal of the umbrella project is to
and management when making decisions regarding biodi- prioritize and characterize the risk that the trade of wildlife
versity conservation, wildlife health and biosecurity, and and wildlife products poses to the US food and agriculture
domestic animal and public health, when wildlife disease is systems and public health. The implication of this research
a critical factor. In an attempt to support interdisciplinary is to inform US regulating agencies of potential wildlife
collaboration, encourage informed decision making, align import risks that may have not been previously considered
language, and limit confusion, the IUCN adopted the and to inform potential risk management and ongoing risk
terminology and framework of the OIE in regard to wildlife communication.
risk analysis.
Case Study: Characterization of the Risk
Conducting the Process (Pathways, Threats, Vulnerabilities, and
Consequences) That the Trade of Wildlife and
The standards and applications of risk analysis are laden Wildlife Products Poses to the US Food and
with jargon, politics, and variability, which often cause
unnecessary confusion and frustration. To begin, risk analy-
Agriculture Systems and Public Health
sis and risk assessment are different. Risk analysis refers to Problem Formulation
the overall process, with its independent components. Risk Because of the shear scope and volume of this issue, we first
assessment is merely a phase of the risk analysis process. conducted an extended “Problem Formulation Exercise”
During the introductory period risk analysis, the initial (initial component of risk analysis) to better understand the
step—and a critical component of risk communication—is issue and prioritize the most important import pathways;
called Problem Formulation. During this step, one should this was followed by three preliminary qualitative risk
do the following: assessments (subsequent component of risk analysis). Step
• Write a general description of the problem, including one, Problem Formulation, consisted of:
why there is a need or opportunity for science-based • A summary of 13 years of US Fish and Wildlife Service
policy or decision making in this case. (USFWS) Law Enforcement Management Information
• Illustrate the problem: draw a picture or diagram to System (LEMIS) data, capturing all declared and unde-
visually represent the issue. clared US wildlife importation records7;
• Identify the “standard” you will follow (in the following • A multi-phased stakeholder engagement survey to char-
case study we will use OIE-IUCN Guidelines highlighted acterize and rank perceptions and priorities surrounding
above) in order to establish what methodology will be this proposed threat;
implemented. • A multidisciplinary stakeholder workshop that reviewed
• Identify pertinent stakeholders—those that will need to results of the aforementioned data and survey results,
be a part of the process, or who the science-based policy and established ranking criteria to prioritize areas of
issue or decision may affect. greatest concern for further assessment.
• Formulate a plan for communicating with and/or inclu- Core partners included advisors from the US Gov-
sion of stakeholders. ernment (DHS, US Geological Survey, Food and Drug
6 SE C T I O N 1 General
Administration, Centers for Disease Control and Preven- is disease based, but it needn’t be. Regardless, it is recom-
tion, US Department of Agriculture [USDA]), as well as mended to start with a list of ranking criteria related to
private agriculture industry representatives, academia, and the threat or hazard of concern under the conditions being
several relevant nongovernmental organizations. Data on considered. For disease, ranking criteria might consist of:
wildlife trade were derived from the “EcoHealth Alliance • Infectivity/transmissibility (ID50 and LD50, Ro)
‘LEMIS’ database”—a compilation of over 20 years of the • Pathogenicity
Convention on International Trade in Endangered Species • Severity such as morbidity, mortality, reproductive
(CITES) and USFWS LEMIS data obtained from the effects, immunosuppression
USFWS via a series of requests over multiple years through • Presence of competent vectors
the Freedom of Information Act. Country disease status • Species susceptibility, risk of crossing species barriers
and disease standards information were obtained through • Economic impacts on species of concern
open access OIE sources such as the World Animal Health • Other ecosystem effects
Information Database (WAHID).8 Data regarding US In this particular analysis, the concern was based upon
agriculture were obtained through several USDA Agricul- “policy and economic” criteria, highlighting diseases of
tural Research Service/Animal and Plant Health Inspection international importance (as defined by OIE listing) being
Service (APHIS)/Foreign Agricultural Service portals, as introduced into the US live animal agriculture system (i.e.,
well as through our advisory team led by Dr. Tracey Dutcher What is the risk of introduction of OIE listed FAD into
(One Health Coordination Office, APHIS). US livestock via the global wildlife trade?). In this case,
Over the 13.5 years examined, wildlife imports to the the implied priority criterion would be transmissibility or
United States included a total of 5,207,420 individually infectiousness (“spreadability”) once introduced, and its
identifiable shipments between January 1, 2000 and August ability to cross species barriers and cause infection and/or
6, 2013. The number of annually declared wildlife shipments illness. It is important to be very specific about the end-
doubled during the period examined, reaching approximately point of concern because this is the equivalent of a research
400,000 declared shipments imported in 2012 alone. These hypothesis in the risk analysis framework. In this case study,
shipments involved a total of 11,033,468,322 individual we were interested in confirmation of at least one “case” of
specimens/animals, plus an additional 977,109,143 kilo- FAD introduction as defined by the US FAD investigation
grams of specimens/animals measured only in weight. The guidelines. Each disease of concern should be evaluated
majority of shipments contained mammals (27%), while via all the ranking criteria and then ranked overall. Often,
the majority of total specimens imported were shells (57%) this is done semi-quantitatively in a spreadsheet, or as a
and tropical fish (25%). decision tree. In our particular case study, we were given
Of the more than 11 billion individual wildlife specimens further guidance by our stakeholders to prioritize FADs of
imported, 27.4% were individually recorded as live upon ruminant livestock.
entry (an annual average of 224.9 million [s = 42.3 million; The real value of this process is for stakeholders to discuss
median = 231.5 million] live animals plus an additional 1.8 which criteria make a disease important in a given scenario,
million kilograms of live animals). Aquatic, amphibian, and before the assessment, in order to again prioritize time
invertebrate species accounted for approximately 50% of and resources on the most crucial issues. Technically, every
these live shipments, mainly imported by the aquatic and high priority hazard must be evaluated further in the risk
pet industries. Reptile, rodent, and bird species destined assessment phase—it is easier to evaluate 5 rather than 100
for the exotic pet trade made up the majority of remaining diseases, and potential hazard lists are often that long. In
live imports.7 our case study, Rift Valley fever (RVF) ranked the highest
Based upon these summary findings and stakeholder pri- priority, both objectively and through the expert elicita-
oritization, three separate Risk Assessments were performed: tion process. The high-risk category also included foot and
1. Risk of introduction of OIE listed foreign animal diseases mouth disease and Crimean-Congo hemorrhagic fever.
(FAD) into US livestock via the global wildlife trade;
2. Risk of introduction of Middle East respiratory syndrome Risk Assessment
(MERS) into US public health via the international In the risk assessment phase, one must ask the questions,
wildlife (camel) trade; “How likely is the hazard to occur?” and “What are the
3. Risk of introduction of OIE listed FAD to US aquacul- consequences if it does occur?” for each priority disease
ture industry via the importation of live aquatic animals identified in the hazard identification phase. The risk
from Asia. assessment phase involves building a representative model
The rest of this chapter will focus on the first case study of the process, collecting data and/or expert opinion, and
illustration. characterizing the outcome in some way. Disease modeling
has recently become all but an entire discipline in itself;
Hazard Identification thus, only the basic premise is highlighted here. A risk
Once pathways of risk are established, the questions “What assessment model is a simplification of the real world and
can go wrong?” and “How can it go wrong?” are posed should help determine the likelihood or probability of
(the core of hazard identification). Usually the discussion adverse health effects associated with hazard exposure. This
CHAPTER 2 Risk Analysis Framework Guidance for Wildlife Health Professionals 7
may be qualitative or quantitative depending on the needs reflects multiple consequence factors including both the
of the users and the amount and type of data available. health of US populations (morbidity and mortality) as well
Often, it also must be informed by expert opinion in the as the economic results from transmission and disease. Spe-
wildlife community due to lack of hard data. There are cifically, this case study analysis was concerned with the con-
scientific methods, such as the Delphi method, among sequences of exposure to the US cattle and swine industries,
others, for the collection and analysis of expert opinion as this risk concern was expressly prioritized by our project
that may be used to add rigor to this process.9 stakeholders. However, consequences of exposure of addi-
Risk assessment is an iterative process as both models and tional animal (e.g., small ruminants) or human populations
data are often refined and updated over time. Usually a brief may be considered in future risk assessments.
qualitative assessment gives a good indication of general risk, Data Sources: For this case study, two main datasets
which allows for the collection and assessment of available were “mined” (i.e., studied) for analysis. First, we combined
data and the likelihood of successful quantitative modeling. USFWS LEMIS data from 2000 to 2013 into a standard-
Quantitative models may be simple, or deterministic, using ized dataset and used these data to extract all declared or
point estimates that usually don’t reflect the range or vari- confiscated live wildlife imports into the United States.
ability of the data. Stochastic models are used to incorporate Second, data on RVF official country status were obtained
uncertainty surrounding point estimates, and involve the from the OIE’s WAHID and Handistatus II portals.8,10
need to match the question and available data with the Approach/Assessment Platform: The general modeling
appropriate tool and method—a more expert modeler approach applied here for risk assessment follows the quali-
should be included in the team if this approach is taken. tative methodology put forth in the IUCN/OIE Guidelines
Often, policy makers want quantitative answers where there for Wildlife Disease Risk Analysis.5 The general format
are no data to support the kind of model that would produce for analyzing risk is the following: Risk = Entry Risk +
the type of specific advice requested. Providing inaccurate Exposure Risk + Consequences.
estimations of the limitations of current results is a major Entry Assessment: A total of 53 species were identified
pitfall in this process. Thus, communicating this potential as meeting the minimum requirements of a medium-risk
mismatch effectively is a large part of risk communication species or higher (17 Artiodactyla, 4 Carnivora, 10 Primate,
between scientists and policy makers. 1 Proboscidea, 3 Perissodactyla, 12 Rodentia, and 6 Bat).
In order to build a risk assessment model, the problem Of the 53 identified species, 20 were imported between
definition needs to be specifically refined, just like that of 2000 and 2013, half of which were Artiodactyla. The top
a scientific hypothesis. six most imported medium- to very-high-risk species made
• Risk assessment question: What is the risk of introduc- up 82.9% of the total number of medium- to high-risk
tion of RVF entering the United States and infecting the individuals imported. These six included the African lion
beef, dairy, and pork industries via trade in live wildlife (Panthera leo; n = 246), springbok (Antidorcas marsupialis;
species based on assessment of trade data collected from n = 134), cheetah (Acinonyx jubatus; n = 121), natal mul-
2000 to 2013? timammate mouse (Mastomys natalensis; n = 100), desert
Under the OIE trade paradigm, the risk assessment warthog (Phacochoerus aethiopicus; n = 48), and impala
is divided into three distinct parts: entry risk (threats), (Aepyceros melampus; n = 45).
exposure risk (vulnerability), and consequences. Entry risk Imports were further refined by country status, so there
involves all steps in the pathways from countries of origin were 381 individuals from 84 shipments from 2000 to 2013
to the US ports of entry (i.e., incoming threats). Exposure of high- to very-high-source risk. Because quarantine in the
involves any steps following entry in which an imported source country could not be confirmed to include vector
animal could potentially expose US populations of animals control, and the majority of shipments were made in less
or people (i.e., US vulnerability to incoming threats). than 3 days, it was assumed that little risk was mitigated
Consequences involve the severity of consequences that are during these factors. Therefore, entry risk was high- to
likely to occur following exposure of US populations (either very-high for n = 381 animals entering the United States
animal or human, to incoming threats). over a 14-year period (2000–2013). This comprised 11
From the initial combination of country disease status species, 5 of which were wild ruminants (187 of the 381
and species host-pathogen status, the risk of exportation imported individuals; 49%). Large carnivores accounted for
and ultimate entry of RVF into the United States can be another 155 of the 381 (40.7%).
increased or reduced by multiple steps along the trade path- Exposure Assessment: In a case where an infectious animal
way, which can be summarized as either shipment or quaran- enters the United States and quarantine measures are
tine factors that contribute to or mitigate risk. The exposure inadequate (e.g., mosquito exposure, subclinical long-term
risk (vulnerability) assessment involves all steps in the path- infectivity, and fomite transmission), the potential spread
way following arrival in the port of entry; these steps in- is high due to the fact that most high-risk imports were
clude transit to US quarantine, US quarantine itself, transit comprised of groups rather than individual animal ship-
to the final destination, and interactions that may occur at ments. This increases the potential spread and complexity of
the final destination between imported wildlife and nearby trace-back during investigation, should a negative scenario
US populations. The consequences portion of the pathway unfold. Quarantine procedures and regulations for wild
8 SE C T I O N 1 General
ruminants are assumed to significantly reduce exposure risk but also potential costs of decisions associated with high
when properly conducted, but there is great uncertainty sur- priority pathogens and their impacts. The idea is to provide
rounding this factor when quarantine occurs at non-USDA scientific input to managers or policy makers about the
facilities, which was found to be a common occurrence. potential costs and benefits of options they are considering,
Further, there is quarantine effectiveness uncertainty regard- or that stakeholders may suggest.
ing diseases such as RVF, the pathogenicity variability of
which we are still working to understand in various wildlife Risk Communication
species. It is often said that risk analysis is an “objective” process. The
Some nonruminant species, such as large, wild carni- reality is that in wildlife and/or disease risk analyses there
vores, are not required to be quarantined upon entry to are often so few data available that the analyst begins, sub-
the United States; others, such as rodents and nonhuman consciously, to substitute value judgments for facts. Indeed,
primates, may be regulated but not screened for RVF. In in assessing the consequences of disease introduction, for
either case, internal domestic shipments may occur after instance, a degree of subjectivity is almost unavoidable.
port of entry—before quarantine—when quarantine is While this may be less true for laboratory settings, it is more
approved for non-USDA facilities. This adds uncertainty likely when assessing disease or environmental risks in free-
to the exposure assessment and may add risk. We found no ranging wildlife. Risk analyses are seldom truly objective,
data to help define potential risk pathways once animals and for this reason transparency in declaring all assumptions
exit the port of entry or quarantine station within the made is essential.11 Most assessments go through several
United States. This represents the greatest gap in informa- iterations, with data collection needs (gaps) highlighted,
tion for this assessment and presents a major opportunity that are then either filled or augmented with gathered expert
for innovation that would help assess the risks outlined or opinion. It is very important not only to cite the source of
exemplified in this assessment. Further, final destination all data, but also to estimate the quality of the data as a
information was not made available for this assessment. contribution to an overall assessment of uncertainty sur-
Therefore, the best potential proxy for the missing pathway rounding results. Thus, risk analysis, although often policy
data above was not available. This represents an opportunity driven, must be a scientifically honest evaluation of what we
to further define risk and fill a major gap in this assessment. know and don’t know. Transparency itself is a commitment
Thus, exposure risk could not be sufficiently categorized to open communication.
with the data available, but exposure risk likely ranged Communicating Uncertainty: Assumptions are what
from low to high depending on the taxonomic order of the we make when we are uncertain. Some assumptions are
imports; however, there was not a large number of risky considered big, others small, depending on the lack of data
importations relative to the large number of overall wildlife or data quality. In these cases, there is usually a gradient of
imports during this time period. Overall, many data gaps evidence, such as expert opinion, proven case studies, or
exist for this portion of the assessment. even local knowledge. There are formal scientific processes
Consequence Assessment: Due to the high morbidity for eliciting expert opinion (such as the Delphi method),
and mortality of the disease in cattle, and the potential and vetted methods to deal with the variability of uncertain
for catastrophic economic loss in both cattle and swine data. The goal of this process is to lay the evidence out logi-
industries, the risk of RVF imports was considered to be of cally and to examine the accompanying level of confidence
low to high likelihood, but whatever level of risk, of high in order to make the best use of existing data, fill important
consequence. gaps, and put responsible bounds around the results. A
risk assessment may sometimes be criticized because many
Risk Management assumptions are made. However, these cases communicate
Risk management is the process of identifying, selecting, the fact that data gaps exist, and great uncertainty is present,
and implementing measures that can be applied to reduce which is important to establish formally in many cases,
the level of risk. Many times these are disease prevention because it allows for discussions regarding how one might
and control strategies, such as vaccination and treatment collect data for an improved evaluation of risk.
of individuals or populations, or personal protective The “Precautionary Principle”: In situations where there
measures, such as wearing gloves and masks for humans is significant scientific uncertainty regarding a risk and
facing zoonotic diseases. The idea is to rerun the model its consequences, such as a cause-and-effect relationship
under different conditions or assumptions to see how the not being fully established, the “precautionary principle”
risk changes in response to intervention actions. Sensitiv- is often invoked. This principle holds that a more cau-
ity analysis—the process of examining the impact of the tious approach should be taken in the face of insufficient
variation in individual model inputs on the model outputs information. In many cases, the precautionary approach
in a quantitative risk assessment—is often performed to has a useful protective effect as the initial response to a
accomplish this. Many times, cost is entered into the equa- potential threat with consequence, especially where valu-
tion as well in order to conduct cost-benefit analyses of able threatened or endangered species—or the release of
different management options. The result is a very powerful infectious diseases—are concerned. On the other hand, too
tool for management authorities to analyze not only risk, much or unnecessary precaution may prevent vital progress
CHAPTER 2 Risk Analysis Framework Guidance for Wildlife Health Professionals 9
in the long term. A transparent discussion of this approach have little evidence-based information for pathogenesis
is recommended. The risk communication strategy should in captive wildlife, or for how transmission may occur
include both more cautious and less cautious solutions for across the species barriers. This is another large data
discussion. gap that affects the consequence assessment and presents
Finally, the risk communication process is essential in further opportunity to support research on both the
helping decision makers to deal with one of the most diffi- ecology and pathogenesis of these agents beyond the
cult problems encountered during the risk analysis process; normal domestic animal realm.
namely, determining what constitutes an “acceptable risk.” There is evidence that there is some level of risk of
Zero risk is seldom, if ever, attainable and some degree of RVF transmission to US livestock from the importation of
risk is unavoidable—this must be stated at the outset. For wildlife species. While the number of imports that are most
example, what is the risk of reintroducing great apes into the likely to provide a risk of RVF transmission are relatively
wild? Can we ever hope to create a situation where there is few compared to the overall large volume of imports, the
zero risk of disease introduction? However, in our passion consequences of a transmission event would be extensive.
and excitement, we may convince managers and funders to Because of this risk, we have recommended investment in
move forward with little regard for potential implications further areas of research and further risk reduction measures.
if this is not discussed up front. On the other hand, health
experts may unnecessarily throw up arbitrary barriers due to Summary
high perceived risk, which is unsupported by a lack of data
and/or great uncertainty surrounding adverse outcomes. Assessing risk is part of the human (and animal) condi-
This discussion intersects with that of the precautionary tion. It is a process by which we learn and change our
principle approach. The goal of risk analysis is to decrease behavior for a more successful future. Risk analysis that is
gridlock, not create paralysis. transparent, logical, and testable is a purposeful method
In the example case study provided here, the statement of conducting this conversation, and—ideally—informing
of “Conclusions and Uncertainty” follows: decisions. Simply, it is the interface between science and
• Overall, with the USFWS LEMIS data available, we can management/policy decision making. It allows for a rela-
make confident statements about viable entry pathways tively quick situation analysis for immediate decision needs,
and volume of trade. These data are limited to declared as well as planning for better, more informed, decisions in
shipments and confiscations. We found no way to the future through the collection of more/better data or the
adequately estimate the illegal wildlife trade. use of more sophisticated tools. In the end, it is the triage
• By using the OIE WAHID, we can infer source risk process of science-based management.
by region but have no way to assess the prevalence in
wildlife or the specific source of animals beyond the Acknowledgments
country of origin and the port of export.
• The largest point of uncertainty in the entry assessment The case study herein was conducted by a large team
surrounds the likelihood that any given animal selected of authors who graciously allowed us to include it as a
for shipment is adequately represented by the country case study in this work. Their names are not included in
status from which it came. the chapter heading due to limitations on authorship in
• We used country status, as reported by the OIE, as this publication format. Although Travis and Smith were
a proxy for source risk. According to experts, there is Co-PIs of the larger project, we recognize authorship-level
major uncertainty surrounding self-reported data on contributions from Peter Sebastian; Shaun Kennedy (Food
many diseases from many countries. System Institute); Tiffany Wolf and Alexander Primus (Uni-
• Surprisingly, for exposure assessment, there is generally versity of Minnesota); Carlos Zambrana-Torrelio, Allison
less data available from which to estimate risk within White, and William Karesh (EcoHealth Alliance). We also
the United States than there is from outside US borders. acknowledge and thank our USG advisory panel for their
The complete lack of formal data on animal movement support of this work: Dr. Tracey Dutcher (USDA, APHIS,
within the United States after entry limits our ability VS); Dr. Jonathan Sleeman (USGS, National Wildlife
to assess the wildlife–livestock interface potential at the Health Center); Dr. Adam Langer (CDC, NZECID); and
endpoint of this trade pathway. Dr. Johnny Braddy (FDA, CFSAN and Chair, Bushmeat
• The acquisition of “importer” data might help tighten Working Group). This project is supported by the US DHS
this assessment slightly but won’t detail the post-entry S&T through a grant awarded by the Food Protection and
transportation methods, the final destination charac- Defense Institute.
teristics, and the purposes of imports (i.e., exposure
of imported animals to humans or other animals).
• Most of the hazards/diseases prioritized by our stake- References
holders were high consequence on either economic or
population morbidity/mortality scales (as confirmed 1. Von Neumann J, Morgenstern O: Theory of games and economic
through stakeholder elicitation above). Many FADs behavior, ed 2, Princeton, 1947, Princeton University Press.
10 SE C T I O N 1 General
2. National Research Council: Risk assessment in the federal govern- analysis, Paris, 2014, OIE. Published in association with the
ment: managing the process, Washington, DC, 1983, The National IUCN and the Species Survival Commission, 24 pp.
Academies Press. 7. Smith KM, Zambrana-Torrelio C, White A, et al: Summarizing
3. Ecological Risk Assessment: https://www.epa.gov/risk/ecological- US wildlife trade with an eye toward assessing the risk of infec-
risk-assessment. (Accessed 20 July 2017). tious disease introduction, Ecohealth 14:29, 2017.
4. OIE: Terrestrial Animal Health Code, Chapter 2.1. http://web 8. OIE World Animal Health Information Database: http://
.oie.int/eng/Normes/mcode/en_chapitre_1.2.1.pdf. (Accessed www.oie.int/animal-health-in-the-world/the-world-animal-health
20 July 2017). -information-system/data-after-2004-wahis-interface/.
5. Jakob-Hoff RM, MacDiarmid SC, Lees C, et al: Manual of pro- 9. Linstone H, Turoff M: The delphi method: techniques and
cedures for wildlife disease risk analysis, 2014. World Organisation applications, Reading, Mass., 1975, Addison-Wesley. ISBN
for Animal Health, Paris, 160 pp. Published in association with 978-0-201-04294-8.
the International Union for Conservation of Nature and the 10. OIE Handistatus II: https://web.oie.int/hs2/report.asp.
Species Survival Commission. 11. MacDiarmid SC, Pharo HJ: Risk analysis: assessment, manage-
6. World Organization for Animal Health (OIE) & International ment and communication, Rev - Off Int Epizoot 22(2):397–408,
Union for Conservation of Nature (IUCN): Kock R, Karesh 2003.
WB, Skerratt L, et al, editors: Guidelines for wildlife disease risk
3
Wildlife Technologies
KAREN BAUMAN
H
istorically many of the technologies used for animals monitor the physiologic responses of pilots, was adapted
have been categorized as being field, agriculture, in the early 1960s as a technique to study small mammal
zoo, or lab based, likely because most have been populations.10,14 The differences between biologging and
adapted from use in humans (medicine), domestic animals, biotelemetry are primarily the means by which data are
or lab animals (biomedical). However, just as the traditional received and stored, but researchers also separate the two
dichotomy between zoo-based staff and field biologists has based on the environments in which they are used.15 In bio-
blurred in the past decade, so too has the application of telemetry, the device carried by the animal is a transmitter
technologies for wildlife become more of a continuum. (sometimes referred to as a tag) that does not store the col-
This philosophical shift, coupled with advances in technol- lected data, but rather continuously transmits it to receiver,
ogy, including the miniaturization of microprocessors and which is typically connected to a computer to automate
integrated data management tools, has led to an exponential data collection. Most receivers are remotely located (meters
growth of technologies that are relevant to veterinarians to kilometers) from the devices, but some must be located
working with wildlife.1,2 close (centimeters), for example, to read radio frequency
This chapter will focus on three types of technologies: identification devices (RFIDs; passive and active types),
(1) biotelemetry and biologging, (2) environmental loggers, such as passive integrated transponder (PIT) tags. Biolog-
and (3) digital imaging. Recent advances in endocrine ging traditionally has been used in marine environments
monitoring, contraception, and diagnostic imaging (radiol- where the radio signals that transmit data for biotelemetry
ogy, computed tomography, magnetic resonance imaging) do not propagate. Data are recorded and stored on the
are discussed elsewhere in this edition (see Chapters 13, device carried by the animal (sometimes referred to as
22, 31, 32), and ultrasound, thermography, and molecular archival or store-on-board loggers). The differences between
technologies have been reviewed recently in previous edi- the two technologies are subtle, and within the past decade
tions of Zoo and Wild Animal Medicine.3–5 there have been new hybrid devices, as well as crossover in
data retrieval techniques.2,15 Because there is now overlap
Biotelemetry and Biologging between the two technologies, for simplicity in this chapter
the term biologging will be used generically to refer to both
Karesh previously reviewed biotelemetry for wildlife veteri- technologies.
narians and provided a thorough description of the equip- Biologging devices may be equipped with a wide range
ment and a basic explanation of the functionality related of sensor types (e.g., heart rate, blood flow, temperature,
to animal location (colloquially referred to as telemetry).6 locomotion).11,16 Devices come in many forms, with most
Although positional estimation of an animal’s location using being custom-made to facilitate them being carried by a
either very high frequency (VHF) radio signal or global diversity of species—from insects to whales (see Cooke et al.,
position system (GPS) technology has obvious importance 2004 for review by order).11 This customization ensures that
to wildlife veterinarians for studies of disease ecology, there the devices are the correct weight and shape for the animal
are several excellent books and reviews available that cover being monitored, collect data for the specified time period,
these aspects of the technology in detail.1,2,7–9 Discussion of and will remain affixed to the animal as required. Weight
biotelemetry in this chapter will focus on its less known uses of the device is a tradeoff between the weight of the battery
for physiologic and behavioral data collection. needed to collect the data over the specified time period and
Biotelemetry and biologging are techniques that allow the body weight of the animal; it is generally recommended
remote measurements of physiologic and behavioral data that the devices not exceed 2%–5% of the body weight
from devices carried by animals.10–13 Biologging was first of the animal.11,17 However, this “rule of thumb” does
developed in the 1940s to study species in marine envi- not take into account other measures of animal welfare,
ronments.13 Biotelemetry, which was originally used to such as changes to behavior, energetic output (especially
11
12 SE C T I O N 1 General
related to increased drag for aquatic and avian species), metabolic rate.33 Subcutaneous devices are used to measure
and discomfort. Animal welfare has been cited as a concern rectal and vaginal temperatures in cattle, and Hoskinson
in biologging studies, with authors asserting that devices measured cloacal temperatures in lorikeets for compari-
should be attached in such a way as not to cause, or at least to son with core temperature.28,34 They can also be used to
minimize, detrimental effects to the individual animal.1,15,18 measure activity and movement.31 Cardiovascular disease
This may be less of a concern in the future because new is a common cause of morbidity and mortality in captive
battery technology is facilitating miniaturization of devices great apes, and several authors have begun to use devices
and some RFIDs are now being equipped with sensors. to monitor cardiovascular parameters in unanesthetized
Biologgers generally range in cost from $100 to $500 per chimpanzees.35 Similar devices have been used in domes-
unit, with some very specialized implantable devices costing tic animals and to study hibernation effects in bears.36,37
$1000 or more. Most devices may be reused, although Lastly, rumen boluses to measure pH may be applicable
they typically have to be sent back to the company and to wildlife species, as well as the ingestable cameras cur-
refurbished. The most significant expense associated with rently used in humans to study intestinal transit times and
biologging is the purchase of the receiving equipment and pathology.28,38
software. Albeit expensive, the purchase may be considered
a long-term institutional investment in much the same way Environmental Loggers
that purchasing an endoscopic equipment or ultrasound
system would be. Data about ambient conditions, as well as information
Physiologic and behavior biologging sensor-equipped about the environment in a habitat, may be useful for
devices can be divided into two attachment categories: addressing research and husbandry questions, as well as pro-
external and internal. In most wildlife species, attachment viding important information to veterinarians and animal
of either type requires anesthesia, but the external devices care staff. Environmental loggers differ from biologgers in
rarely require surgery. External attachments are less expen- that they are intended to be used within the environment
sive than internal devices equipped with similar sensors and (e.g., mounted to a wall or a probe inserted in the soil to
include an array of devices that can be glued onto skin, measure moisture) and not carried by an animal.
shells, or feathers; sutured or strapped on (such as halter- Most environmental loggers are used in industry so they
type monitors); worn (e.g., collars, ear tags, or bands); and are available in a great diversity of sizes and types. Those
sat upon (eggs). Although unusual in wildlife applications, that may be of interest for monitoring changes that occur
a halterlike biotelemetry belt has been used to measure in and around an animal’s habitat or conditions during
heart rate and respiration in wildebeest (Connochaetes tau- animal shipments include: temperature; humidity; light;
rinus).19 Other less common applications include the use water flow, level, turbidity, and salinity; barometric pres-
of acoustic recording devices on collars to record chewing sure; soil moisture; carbon dioxide; acoustic sound pressure
and vocalizations and miniature video cameras attached to and decibels; and wind speed. Environmental loggers can
the ventral feathers of birds and collars of mammals.20–23 measure a single condition or combinations of up to three
Accelerometers are now often used in collars and glued- variables. It is important to note that not all environmental
on devices to study behavior, activity levels, or circadian loggers are battery powered (some require AC power), and
and/or movement patterns.24–26 Passive- and active-type all are rated for specific environmental conditions (e.g.,
RFID tags are commonly used in domestic animal species indoor only). Most are designed to store data until retrieved
to monitor feeding bouts, feeding and drinking amounts, for download, but WiFi and mini network capabilities are
locomotion, and activity, as well as estrus behavior and becoming more common. Because environmental loggers
general health.27–29 In elephants, active RFIDs have been are typically available “off the shelf,” they are often less
used to study social affiliations, activity, and use of exhibit expensive than biologgers, averaging approximately $150
spaces.29,30 per unit. In most cases they are “standalone,” so the only
Most internal devices were developed for human or additional cost is for software and potentially a download
biomedical research purposes and tend to be more expen- cable. There are several important factors that must be
sive, technologically intensive, and, in some cases, invasive considered when using environmental data loggers. The
compared with external devices. Internally implantable logger must be capable of recording data within the range
equipment, such as intraperitoneal and subcutaneous you believe possible, plus an appropriate buffer on either
devices, as well as those that are sutured to blood vessels side. For example, a temperature logger with a range of
and organs, requires surgical placement. Until recently, 10–37.7°C (50–100°F) would work well for warm water
core body temperature could be measured only with intra- jellyfish but would likely be unsuitable for cold water jellies,
peritoneal devices or devices sutured to blood vessels, but which thrive in water 15°C (59°F) or colder.39 Another
recently miniaturized devices with thermographic sensors important consideration is the logging interval, which must
have been used to compare intraperitoneal, subcutaneous, be selected carefully because it affects battery life and data
and intramuscular temperatures in antelope.31,32 Rey also storage. More frequent sampling results in more data to
placed rabbits with multiple, similarly placed temperature be stored (and analyzed later!) and drains battery power
devices and an accelerometer into a respirometer to measure more quickly than longer sampling intervals. In addition,
CHAPTER 3 Wildlife Technologies 13
7. Millspaugh J, Marzluff JM, editors: Radio tracking and animal 28. Caja G, Castro-Costa A, Knight CH: Engineering to support
populations, San Diego, 2001, Academic Press. wellbeing of dairy animals, J Dairy Res 83(02):136–147,
8. Silvy NJ: The wildlife techniques manual: volume 1: research. 2016.
Volume 2: management 2-vol. Set (vol 1). Baltimore, 2012, John 29. Wall J, Wittemyer G, Klinkenberg B, et al: Novel opportunities
Hopkins University Press. for wildlife conservation and research with real-time monitoring,
9. Tomkiewicz SM, Fuller MR, Kie JG, et al: Global positioning Ecol Appl 24(4):593–601, 2014.
system and associated technologies in animal behaviour and eco- 30. Scott NL, Hansen B, LaDue CA, et al: Using an active Radio
logical research, Philos Trans R Soc Lond, B, Biol Sci 365(1550): Frequency Identification Real-Time Location System to remotely
2163–2176, 2010. monitor animal movement in zoos, Anim Biotelemetry 4(1):16,
10. Kenward RE: Historical and practical perspectives. In Millspaugh 2016.
J, Marzluff JM, editors: Radio tracking and animal populations, 31. Asa CS: Biotelemetry applications for captive animal care and
San Diego, 2001, Academic Press, pp 3–12. research. Bethesda, The American Association of Zoological
11. Cooke SJ, Hinch SG, Wikelski M, et al: Biotelemetry: a mecha- Parks and Aquariums, 1991.
nistic approach to ecology, Trends Ecol Evol (Amst) 19(6):334–343, 32. Rey B, Fuller A, Hetem RS, et al: Microchip transponder
2004. thermometry for monitoring core body temperature of antelope
12. Rutz C, Hays GC: New frontiers in biologging science, The Royal during capture, J Therm Biol 55:47–53, 2016.
Society, 2009. 33. Rey B, Halsey LG, Hetem RS, et al: Estimating resting metabolic
13. Ropert-Coudert Y, Beaulieu M, Hanuise N, et al: Diving rate by biologging core and subcutaneous temperature in a
into the world of biologging, Endanger Species Res 10:21–27, mammal, Comp Biochem Physiol A Mol Integr Physiol 183:72–77,
2009. 2015.
14. Mech LD: Telemetry as a technique in the study of predation, 34. Hoskinson C, McCain S, Allender M: Comparison of body
J Wildl Manage 492–496, 1967. temperature readings between an implantable microchip and a
15. Cooke SJ: Biotelemetry and biologging in endangered species cloacal probe in lorikeets (Trichoglossus haematodus sp), J Avian
research and animal conservation: relevance to regional, national, Med Surg 28(4):355–356, 2014.
and IUCN Red List threat assessments, Endanger Species Res 35. Magden ER, Sleeper MM, Buchl SJ, et al: Use of an implant-
4(1–2):165–185, 2008. able loop recorder in a chimpanzee (Pan troglodytes) to monitor
16. Wilson AD, Wikelski M, Wilson RP, et al: Utility of biological cardiac arrhythmias and assess the effects of acupuncture and
sensor tags in animal conservation, Conserv Biol 29(4):1065– laser therapy, Comp Med 66(1):52–58, 2016.
1075, 2015. 36. Laske TG, Garshelis DL, Iaizzo PA: Big data in wildlife research:
17. Hawkins P, Morton D, Bevan R, et al: Husbandry refinements remote web-based monitoring of hibernating black bears, BMC
for rats, mice, dogs and non-human primates used in telemetry Physiol 14(1):13, 2014.
procedures, Lab Anim 38:1–10, 2004. 37. von Borell E, Langbein J, Després G, et al: Heart rate variability
18. Wilson RP, McMahon CR: Measuring devices on wild animals: as a measure of autonomic regulation of cardiac activity for
what constitutes acceptable practice? Front Ecol Environ 4(3): assessing stress and welfare in farm animals—a review, Physiol
147–154, 2006. Behav 92:293–316, 2007.
19. Laubscher LL, Hoffman LC, Pitts NI, et al: Validating a human 38. Waghmare PV, Panchal CV, Poul BN: Swallowable wireless cap-
biotelemetry system for use in captive blue wildebeest (Con- sular endoscopy: a novel breakthrough in the biomedical industry
nochaetes taurinus), Zoo Biol 34(4):321–327, 2015. and future progress, Int J Pharm Sci Res 4(11):4133–4140,
20. Lynch E, Angeloni L, Fristrup K, et al: The use of on-animal 2013.
acoustical recording devices for studying animal behavior, Ecol 39. AZA Aquatic Invertebrate TAG: Jellyfish care manual. Silver
Evol 3(7):2030–2037, 2013. Spring, Association of Zoos & Aquariums, 2013.
21. Bluff LA, Rutz C: A quick guide to video-tracking birds, Biol 40. Owen MA, Swaisgood RR, Czekala NM, et al: Monitoring stress
Lett 4(4):319–322, 2008. in captive giant pandas (Ailuropoda melanoleuca): behavioral and
22. Rutz C, Troscianko J: Programmable, miniature video-loggers for hormonal responses to ambient noise, Zoo Biol 23:147–164,
deployment on wild birds and other wildlife, Methods Ecol Evol 2004.
4(2):114–122, 2013. 41. Barber JR, Crooks KR, Fristrup KM: The costs of chronic
23. Moll RJ, Millspaugh JJ, Beringer J, et al: A terrestrial animal- noise exposure for terrestrial organisms, Trends Ecol Evol (Amst)
borne video system for large mammals, Comput Electron Agric 25:180–189, 2010.
66(2):133–139, 2009. 42. Laiolo P: The emerging significance of bioacoustics in animal
24. Nathan R, Spiegel O, Fortmann-Roe S, et al: Using tri-axial species conservation, Biol Conserv 143:1635–1645, 2010.
acceleration data to identify behavioral modes of free-ranging 43. Nazir S, Newey S, Irvine RJ, et al: WiseEye: next generation
animals: general concepts and tools illustrated for griffon vul- expandable and programmable camera trap platform for wildlife
tures, J Exp Biol 215(6):986–996, 2012. research, PLoS ONE 12(1):e0169758, 2017.
25. Mosser AA, Avgar T, Brown GS, et al: Towards an energetic 44. Krukowski M: Automatic determination of body condition score
landscape: broad-scale accelerometry in woodland caribou, of dairy cows from 3D images. Skolan för datavetenskap och
J Anim Ecol 83(4):916–922, 2014. kommunikation, Kungliga Tekniska högskolan; 2009.
26. Kozlowski C, Bauman KL: Use of Fitbit® for studying activity in 45. London GD, Bauman KL, Asa CS: Time-lapse infrared videog-
fennec fox (Vulpes zerda), Unpublished manuscript, 2017. raphy for animal behavior observations, Zoo Biol 17:535–543,
27. Maselyne J, Saeys W, De Ketelaere B, et al: Validation of a High 1998.
Frequency Radio Frequency Identification (HF RFID) system 46. O’Donoghue P, Rutz C: Real-time anti-poaching tags could help
for registering feeding patterns of growing-finishing pigs, Comput prevent imminent species extinctions, J Appl Ecol 53(1):5–10,
Electron Agric 102:10–18, 2014. 2016.
CHAPTER 3 Wildlife Technologies 15
47. Gonzalez LF, Montes GA, Puig E, et al: Unmanned Aerial 49. Chabot D, Francis CM: Computer-automated bird detection
Vehicles (UAVs) and artificial intelligence revolutionizing and counts in high-resolution aerial images: a review, J Field
wildlife monitoring and conservation, Sensors (Basel) 16(1):97, Ornithol 87(4):343–359, 2016.
2016. 50. Whitham J, Miller L: Using technology to monitor and improve
48. Christie KS, Gilbert SL, Brown CL, et al: Unmanned aircraft zoo animal welfare, Anim Welfare 25(4):395–409, 2016.
systems in wildlife research: current and future applications of 51. Pimm SL, Alibhai S, Bergl R, et al: Emerging technologies to
a transformative technology, Front Ecol Environ 14(5):241–251, conserve biodiversity, Trends Ecol Evol (Amst) 30(11):685–696,
2016. 2015.
4
International Sample Movement:
Overview of Convention on
International Trade in Endangered
Species of Wild Fauna and Flora and
Selected National Regulations
CHRIS WALZER
T
he collection of biological samples is central to wild- requiring in most cases the capture and anesthesia of the
life health, conservation, and environmental studies. targeted species, whereas the collection of, for example,
The acquisition and processing of biological wildlife fecal samples can occur noninvasively. Similarly, the collec-
samples is, in most cases, a prerequisite for establishing a tion of hair, urine, feathers, shed skin, saliva, and eggshells
diagnosis. Furthermore, information extracted from biologi- can be performed noninvasively and, in many instances,
cal samples can be instrumental in shaping conclusions and without actually observing the animals.1 It is important to
guiding policy. The sampling procedure across disciplines point out that sampling live and dead wildlife requires a
can be broken down into several distinct steps: (1) acquisi- profound understanding of the inherent risks involved to
tion of the sample, (2) collection/recording of linked sample the sampled animal and the investigator. Risk mitigation
metadata, (3) initial storage of the sample, (4) transport of measures, such as the use of adequate personal protective
the sample, (5) processing of the sample, (6) final storage of equipment (PPE), safe anesthetic protocols, and animal
the sample, and (7) sharing the sample and/or information welfare legislation are but some of the measures to consider.
from the sample. All steps in this process are potentially A large and diverse number of sampling guidelines and
regulated and restricted by national and international leg- recommendations are available and can be used as a basis for
islation and constrained by logistical challenges. Consider- developing a specific sampling procedure.2,3 Certain species,
ing these individual steps while referring to the respective such as nonhuman primates and bats, warrant a heightened
legislation will provide a solid framework for a sampling appreciation and consideration of PPE measures during
plan. Within these individual steps, various options and sampling and subsequent processing due to the potential
constraints can be identified and should be carefully consid- exposure to life-threatening pathogens, such as Ebola virus,
ered at the outset when establishing the sampling protocols. Cercopithecine herpes-1 (B virus), and rabies.4,5
Although this discussion on sampling is pertinent to various Consistently linking individual samples with their
fields and types of samples, it will focus on samples from respective unique metadata is an essential prerequisite to
wildlife. guarantee the effective future use of the sample. Only the
Before samples can be collected, investigators must combination of adequately collected, processed, stored, and
ascertain that all necessary permits for the actual collection annotated samples will allow the generation of meaning-
process have been requested and approved. This can include ful results. Various international initiatives are ongoing
a multitude of permits such as, but not limited to: research to streamline and harmonize sample metadata use (e.g.,
permits, access permits, and working permits. The type of MIABIS: Minimum Information About BIobank data
sample required most often defines the approaches used. Sharing) and can be used as a guideline.6
On the one hand, the acquisition of blood samples from Sample storage varies widely in respect to the type of
live animals in considered an invasive form of collection, sample collected and the subsequent analysis to be performed.
16
CHAPTER 4 International Sample Movement 17
Feathers used for DNA extraction in a genetic study can fall under the regulatory terms of this agreement.10 This
simply be stored in dry paper envelopes; sex hormones includes blood and its derivatives, hair, skin, tissue, and
remain stable when chilled. Although nucleic acid from extracted DNA. In contrast, most countries, but not all,
blood samples is easily captured and stored for years on FTA consider fecal samples to be wildlife byproducts that are
filter cards (Whatman FTA, Sigma-Aldrich Handels Gmbh, exempt from CITES permitting. The first step in applying
Vienna, Austria), blood samples and tissues investigated for for a CITES permit is to ascertain whether the species or
viral RNA must be processed immediately. These samples subspecies in question is covered by the CITES convention
must be stored in an RNA stabilization solution, which and to determine in which CITES appendix the species
stabilizes and protects cellular RNA (e.g., RNAlater Fischer or subspecies is regulated (www.cites.org/eng/resources/
Scientific—Austria GmbH, Vienna, Austria) and in many species.html). Appendix I includes species threatened
cases eliminates the need for liquid nitrogen, depending with extinction, and consequently the trade (and scientific
on the environmental temperature.1,7 In addition to the exchange) in specimens from these species is permitted
type of sample collected, sample storage must be carefully only in exceptional circumstances. Appendix II includes
considered because it potentially introduces important species not necessarily threatened with extinction but in
additional downstream decision points. An alternative which trade must be controlled to avoid overexploitation.
approach to wildlife sampling is to process all samples on Finally, Appendix III contains species that are protected in
site, in-country. This eliminates the international sample at least one country. Depending on the applicable listing,
export process, greatly speeding up diagnostic turnaround the permitting process varies. For Appendix I species an
time. In addition, in-country processing facilitates sustain- import permit issued by the management authority of the
able knowledge and technology transfer and in-country state of import is required first, whereas for Appendix II
data availability. Novel portable diagnostic systems such as species an export permit or reexport certificate issued by the
smartphone-powered quantitative polymerase chain reac- management authority of the state of export or reexport is
tion (PCR; Biomeme two3, Philadelphia, PA, USA) and required initially. A comprehensive and constantly updated
nanopore DNA sequencing (MinION, Oxford Nanopore overview is available online (https://cites.org/eng/disc/
Technologies, Oxford, UK) enable the detection of specific how.php). Although CITES is legally binding in national
genetic material from pathogens and hosts in remote field states, implementation can vary in relation to the specific
settings.8,9 domestic measures adopted for that purpose. Additional
Most readers at one stage or another in their career will international (EU) and national legislation could regulate
have witnessed the surprise arrival of inadequately shipped and limit sample movement. In the United States, for
biological samples in unmarked soggy cardboard boxes. example, the various regulatory mechanisms for migratory
Nonetheless, it is evident that once samples are to be trans- birds must be considered (see: http://www.fws.gov/birds/
ported, a plethora of legislation, rules, and regulations need policies-and-regulations.php; and http://ec.europa.eu/
to be considered and strictly adhered to. Legislation, and environment/cites/info_permits_en.htm). It is therefore
more importantly its implementation, can vary widely be- absolutely essential to contact the national management
tween nations, so the country-specific rules and regulations authority of the respective state(s) (e.g., United States: the
should be determined before sample collection. However, United States Fish and Wildlife Service [USFWS] in the
for the sake of this chapter, we focus on sample movement European Union and most other countries the respective
into and within the United States and European Union. ministries of environment).
At the most basic level, the investigator must ensure that
prior to shipment of samples the respective valid export Veterinary Import Permits
permits from the country of origin and the valid import
permits from the receiving country have been obtained. In addition to CITES and national requirements for the
The types of permits required fall into several categories movement of wildlife samples, respective so-called veteri-
and vary in accordance with the type of sample, species, nary import permits from the national veterinary and/or
and mode of transport. agriculture departments must be obtained (United States:
US Department of Agriculture’s Animal and Plant Health
Inspection Service; European Union: respective Ministries of
Convention on International Trade Health and Agriculture). The objective of veterinary import
in Endangered Species of Wild Fauna permits is to protect livestock or agriculture from materials
and Flora that may pose a threat. Veterinary permits are needed for
a wide variety of materials derived from animals or source
The Convention on International Trade in Endangered materials that have been exposed to animals. Materials that
Species of Wild Fauna and Flora (CITES) is an international require a permit include, among others, animal tissues,
agreement among 183 governments ensuring that the blood, cells or cell lines, fecal samples, RNA/DNA extracts,
international trade in specimens of wild animals and plants hormones, and microorganisms, including bacteria, viruses,
does not threaten their survival. Although scientific biologi- protozoa, and fungi. In some countries, veterinary export
cal samples are in the majority of cases not traded, they fully permits for samples are also required.
18 SE C T I O N 1 General
Be aware that the veterinary import process can be very The European Union ratified the protocol (Regulation
dynamic, and requirements will change at short notice (EU) No. 511/2014) in 2014, and the subsequent Regula-
as the status of animal diseases reported to the World tion (EU) 2015/1866 came into force in November 2015,
Organization for Animal Health (OIE) changes. Veterinary laying down detailed rules and best practices in implement-
requirements for the import of wildlife samples vary widely ing Reg. 511/2014. However, implementation in several
with respect to the species and the country of origin. In member states is currently still lacking. Obligations and
general, the import of ungulate, equid, and bird samples can implementation vary significantly among the signatories,
be extremely problematic, if not impossible. Subsequently, but using the EU as a guideline, users of genetic material
specific restrictions on the type, condition, and approved and traditional knowledge must exercise due diligence to
preservation of the sample will apply. Approved preservation ascertain that: (1) the genetic resources and traditional
can include, among many others: (1) heating to a certain knowledge used have been accessed in accordance with
temperature for a determined time period, (2) immersion in applicable access and benefit-sharing legislation or regula-
formalin or other preservatives, and (3) irradiation. National tory requirements and (2) benefits are fairly and equitably
implementation and requirements will vary significantly. In shared on mutually agreed terms and in accordance with
the United States, irradiation must be performed under any applicable legislation or regulatory requirement. To
the direct supervision of the National Veterinary Services fulfill these obligations, parties must issue a permit or its
Laboratory or the Foreign Animal Disease Diagnostic equivalent at the time of access as evidence that access to
Laboratory (FADDL), Plum Island, NY. In the European genetic resources was based on prior informed consent and
Union, respective national entities supervise this process. It that mutually agreed terms were established. The parties
is important to be aware of the potential negative effects, must make information on the permit or its equivalent
such as deterioration or destruction of genetic material, available to the ABS Clearing-House for the constitution
from the use of the prescribed preservatives and irradiation. of the internationally recognized certificate of compliance.
In some countries, additional requirements can apply The first internationally recognized certificate of compli-
to specific species. For example, in the United States, ance was issued on October 1, 2015 by India’s National
all samples originating from nonhuman primates entail Biodiversity Authority, the competent national authority
special restrictions, additional permits, and health reporting under the Nagoya Protocol, granting access to ethnome-
requirements from the Department of Health and Human dicinal knowledge of the Siddi community from Gujarat
Services and the Centers for Disease Control and Preven- to a researcher affiliated with the University of Kent in the
tion.11 It is highly recommended to contact the respective United Kingdom.14
authorities well in advance of the planned import to discuss Although implementation of the protocol is still lacking
and clarify the import process. in numerous countries, it is also clear that some countries
are strictly adhering to the protocol (e.g., Germany) and
Nagoya Protocol that major granting agencies are already requesting an
internationally recognized certificate of compliance at the
The Nagoya Protocol on Access to Genetic Resources and time of grant submission. It is only a matter of time before
the Fair and Equitable Sharing of Benefits Arising From reputable peer-reviewed journals will also require these
Their Utilization (ABS) to the Convention on Biological certificates prior to publishing results that include genetic
Diversity is a supplementary agreement to the Convention data from wildlife.
on Biological Diversity. The protocol provides a transparent
legal framework for the effective implementation of the fair Packaging and Labeling Samples
and equitable sharing of benefits arising out of the utilization
of genetic resources. The protocol aims to prevent biopiracy A key requirement for a successful shipment is the correct
(i.e., commercial exploitation of biological compounds or choice of packaging suitable to the type of sample to be
genetic sequences by a technologically advanced country shipped and the conditions that may be encountered along
or organization without obtaining consent or providing the route. Many transported samples and their respective
fair compensation to the source country and peoples). The preservatives and additives constitute a dangerous material
Nagoya Protocol on ABS was adopted on October 29, (HAZMAT in the United States) that can potentially inflict
2010 in Nagoya, Japan and came into force on October harm to persons or property and damage to the environ-
12, 2014.12 Although the previously mentioned permits ment, the means of transport used, or to other goods.
regulate the movement of physical samples, the ABS applies The Committee of Experts on the Transport of Dangerous
to genetic resources over which states exercise sovereign Goods of the United Nations Economic and Social Council
rights and to traditional knowledge associated with genetic (ECOSOC) has developed guidelines that assign a four-
resources (traditional knowledge). It is important to note digit code (UN number) to the most common dangerous
that, although some 100 parties have signed and ratified the goods. UN 2814 denotes Infectious Substances, Category
protocol, numerous countries have, to date, not ratified or A, which can cause disease in humans or in both humans
signed (of particular note, the United States) the protocol.13 and animals, whereas UN 2900 is assigned to Infectious
CHAPTER 4 International Sample Movement 19
2. FAO (Food and Agriculture Organization of the United Nations): 11. Centers for Disease Control and Prevention (CDC): Questions
Wild Birds and Avian Influenza. An introduction to applied field and Answers for Importers on the Regulations for the Importa-
research and disease sampling techniques. Manual. In Whitworth tion of Nonhuman Primates (42 Code of Federal Regulations
D, Newman SH, Mundkur T, et al, editors: FAO animal produc- [CFR] Part 71.53), 2017. Retrieved June 14, 2017, from https://
tion and health manual, Rome, 2007, FAO, p 120. www.cdc.gov/importation/laws-and-regulations/nonhuman
3. Kirmaier A, Diehl W, Johnson WE: Acquisition and processing -primates/nprm/qa-importers.html.
of nonhuman primate samples for genetic and phylogenetic 12. Convention on Biological Diversity: About the Nagoya Protocol,
analyses, Methods 49:5–10, 2009. 2010. Retrieved June 14, 2017, from https://www.cbd.int/abs/
4. WHO (World Health Organization): WHO Guide for Rabies about/default.shtml/.
Pre and Post Exposure Prophylaxis in Humans, 2013. Retrieved 13. Convention on Biological Diversity: Parties to the Nagoya Pro-
June 14, 2017, from http://www.who.int/rabies/WHO_Guide tocol, 2017. Retrieved June 14, 2017, from https://www.cbd.int/
_Rabies_Pre_Post_Exposure_Prophylaxis_Humans_2013.pdf. abs/nagoya-protocol/signatories/.
5. Centers for Disease Control and Prevention: Ebola (Ebola Virus 14. Convention on Biological Diversity, UNEP: The first interna-
Disease). Personal Protective Equipment (PPE), 2016. Retrieved tionally recognized certificate of compliance is issued under the
June 14, 2017, from https://www.cdc.gov/vhf/ebola/healthcare Nagoya Protocol on Access and Benefit-sharing, 2015. Retrieved
-us/ppe/index.html. 2 p, from https://www.cbd.int/doc/press/2015/pr-2015-10-07
6. MIABIS: Minimum Information About BIobank data Sharing -abs-en.pdf.
(version 2.0), 2017. Retrieved June 14, 2017, from https:// 15. Weisman W, Smith K, Smith B, et al: PREDICT Operating Pro-
github.com/MIABIS/miabis/wiki. cedures: Packing and Shipping Biological Samples, 2016. http://
7. Tworoger SS, Hankinson SE: Collection, processing, and storage www.vetmed.ucdavis.edu/ohi/local_resources/pdfs/guides/
of biological samples in epidemiologic studies: sex hormones, predict-sop-packing-shipping-biological-samples-2016.pdf.
carotenoids, inflammatory markers, and proteomics as examples, 16. International Air Traffic Association (IATA): Provisions for
Cancer Epidemiol Biomarkers Prev 15:1578–1581, 2006. Dangerous Goods Carried by Passengers or Crew (Subsection
8. Marx V: PCR heads into the field, Nat Methods 12:393–397, 2.3). Dangerous Goods Regulations, 2017. https://www.iata.org/
2015. whatwedo/cargo/dgr/Documents/passenger-provisions-table
9. Quick J, Loman NJ, Duraffour S, et al: Real-time, portable -23A-en.pdf. 2.
genome sequencing for Ebola surveillance, Nature 530:228–232, 17. International Air Traffic Association (IATA): Dangerous Goods
2016. Regulations (DGR), 2017. Retrieved June 14, 2017, from http://
10. Convention on International Trade in Endangered Species of www.iata.org/publications/dgr/Pages/index.aspx.
Wild Fauna and Flora (CITES): What is CITES? 2017. Retrieved
June 14, 2017, from https://www.cites.org/eng/disc/what.php.
5
A Practical Guide for Statistics in
Wildlife Studies
FRANCISCO OLEA-POPELKA AND LAURA ELIZABETH ROSEN
21
22 SE C T I O N 1 General
interpreted properly. In simple terms the selection of a information bias may result when a measuring device does
study (and analysis to be conducted) depends on the study not accurately record the true value of a parameter, for
goal and specific research question. The investigator must example, a field device or laboratory technique (e.g., lactate,
be aware of the limitations, strengths, and purpose of each pH, or optical density values from an enzyme-linked immu-
study type and choose a study design that meets the goals of nosorbent assay) that does not provide accurate readings.
the proposed research. Wildlife researchers could have dif- Confounding bias occurs when a factor (known as the con-
ferent goals, such as (1) describing an outcome (e.g., health founder) is not included in the analysis and this “missing
event, clinical case, or animal management in a facility) or factor” is associated with both the outcome of interest and
(2) estimating the prevalence of a disease or the distribu- another factor that the investigator analyzed. Thus, when
tion of physiologic parameters from blood samples (e.g., confounding bias is present, conclusions may be incorrectly
pH or lactate values). In these scenarios, case reports, case drawn regarding associations between an evaluated factor
series, descriptive reports, or surveys (or census) provide a and an outcome, when in fact, another unmeasured factor is
variety of study options to choose from. When the goal influencing the outcome. For example, say an investigator is
of a study is (3) to compare interventions such as drug evaluating the impact of location (geography) on prevalence
combinations for immobilization, vaccines, or screening/ of a certain disease in a species. Age is known to be associ-
diagnostic tests, physiologic parameters, or risk factors for ated with prevalence of this disease, and assume that 70%
a disease among different groups of animals, analytic studies of animals in location A are adults, compared with only
including experiments (e.g., laboratory or controlled field 15% of animals in location B. If age is not included in the
trials) or observational studies such as cross-sectional, cohort, analysis, then the location results could be affected by con-
and case-control studies are common studies used in this founding bias, where the analysis ignores a factor (age) that
field. Keep in mind that cross-sectional studies are not is both associated with the outcome (disease prevalence) and
appropriate if the goal is to obtain disease incidence because another factor (location). There are several ways to prevent
the outcome and factor are evaluated at a single point in confounding bias during the study design and also means
time; case-control studies are not appropriate if the goal to adjust the analysis to control for potential confounding
is to determine the probability (risk) of disease in a given factors described elsewhere.1,5 In this chapter, common and
population, because the investigator selects the “cases”; and robust statistical methods to adjust results for potential con-
cohort studies are not the best option if the outcome of founding factors are described in Q9 and Fig. 5.1.
interest is rare, because the investigator would need to wait
a considerably long time to identify just a few rare cases in
the study population. Q4: How Many Animals (or Samples)
Do I Need for My Study?
Q3: What May Affect the Validity When estimating disease prevalence or the average value of a
of My Study? continuous outcome (e.g., blood pressure in mm Hg, lactate
values in mM/L), the (1) level of confidence, (2) precision,
Study validity has been described as the absence of a sys- and (3) expected variability in your data will determine the
tematic error (bias) in results.1 The three general types of number of animals/samples required. For example, to con-
biases that may negatively impact the validity of wildlife duct a study with 95% confidence to estimate that the dis-
studies are (1) selection bias, (2) information bias, and (3) ease prevalence is approximately 15% with a precision of
confounding bias. A detailed review of validity and bias in 2% will require a larger sample size as compared with the
veterinary studies may be found in other texts.1,5 In brief, same study with a 6% precision (the variance1,3 in this case
selection bias occurs when the selected study subjects are would be the prevalence times [1-prevalence], thus 0.15 ×
systematically different from those animals in the target 0.85). For analytic studies when the goal is to compare an
population5 or, in other words, when animals selected for outcome between groups, the study should be planned to
the study have different characteristics than those animals have at least 80% power. The investigator must clearly in-
not selected for the study.5 One common example occurs dicate the magnitude of the difference to be detected rather
when wildlife researchers are restricted to have access only than just saying higher or lower. The magnitude of the dif-
to individuals of certain sex and age, and thus in observa- ference to be detected is key in determining the number of
tional studies this scenario may result in a study popula- animals/samples needed; the greater the difference to be de-
tion not representative of the source (and target) population tected, the fewer animals/samples needed, and vice versa,
to which the investigator desires to extrapolate the study the smaller the difference to be detected among groups, the
results. Information bias occurs when the data recorded on larger the sample size needed. Thus a study with 80% power
the outcome or factors are inaccurate, for example, when to detect a difference of 5% in disease prevalence between
a screening/diagnostic test does not correctly classify the females and males and declared statistically significant will
disease or infection status1 of an individual due to the lack require a considerably larger number of females and males
of test sensitivity (causing false negative results) or lack than the same study to detect a difference of 35% between
of test specificity (causing false positive results). Similarly, females and males. Equally, a study designed to detect a
CHAPTER 5 A Practical Guide for Statistics in Wildlife Studies 23
• Figure 5.1 Multivariable analyses options to account for the effect of multiple factors on the outcome
of interest. *Technically, the residuals should be normally distributed, but often, a quick assessment can
be done by evaluating the distribution of the outcome data.
drop in blood pressure of 60 mm Hg after a drug is applied S-PLUSe, EpiToolsf, among others) provide user-friendly
will require a considerably smaller sample size as compared platforms to perform these calculations allowing, if neces-
with a study designed to detect a drop in blood pressure of sary, for adjustment of sample sizes for small populations,
20 mm Hg. Ideally, the difference (hypothesis) to be tested lack of sensitivity and specificity of screening/diagnostic
in any analytic study should represent a clinically, biological- tests, and lack of independence (clustering) when present.
ly, or epidemiologically relevant and meaningful difference.
small sample sizes, which include counts (e.g., 2/6) rather different tests applied to the same blood sample) may be
than using proportions and 95% confidence intervals, and entered in a separate column indicating different sampling
using the median, quartiles (Q1 and Q3), and ranges as points or different tests (0, 1, 2, 3, etc.). All of the collected
opposed to the mean, standard deviation, and correspond- information for that particular individual (e.g., age, sex,
ing 95% confidence interval for continuous outcomes. The treatments, blood pressure, diseased, or nondiseased status)
median is a more stable estimate of central tendency in these should be entered in different columns, with columns con-
scenarios because the mean may be impacted by extreme taining only information relating to one factor or animal/
observations.3,4 In addition, when comparing groups, it is sample characteristic (you may add as many columns as
strongly recommended to use nonparametric statistical tests necessary). Letters or numbers may be used to codify the
designed to analyze data arising from studies with small data, and this greatly facilitates data analysis. For example, if
sample sizes or when data do not meet the statistical test there are different groups in your data set (e.g., two different
assumptions.3 Of course, what is considered a small sample treatment groups), it is best to include the group as its own
size has been the subject of some debate, but depending on column rather than separate sheets for different treatment
the statistical test required, some authors3 consider studies groups (0, 1 or A, B; see Fig. 5.2B). Consistency: Use the
with n < 20 or n < 25 as candidates for nonparametric same unit (e.g., kilograms or pounds) for all measurements.
analyses, whereas others2 consider studies with n < 30 as Record factors using a consistent format (e.g., DD-MM-
candidates for nonparametric analysis. The most important YYYY for date) in every row. For nonnumerical informa-
caution to consider in studies with relatively small sample tion, it is best to use the exact same word or phrase rather
sizes is to avoid generalizing and/or extrapolating results than multiple versions of the same idea (e.g., “left lateral”
to a larger population if the data in fact are not repre- rather than “left,” “L,” “left lateral,” and “L lat”). Similarly,
sentative of a larger population. Rather, the investigator make sure that spelling and letter case for names, drugs,
should acknowledge the study limitations and discuss the etc. are identical across rows. Conciseness: Do not enter
clinical, biological, or epidemiologic relevance of results any unnecessary information in a column. For example,
and highlight the strengths, novelty, and or contribution a numerical column (e.g., temperature) should only have
of their data to the field. In fact, studies with small sample numbers, no letters. The units °C or °F may be in the title of
sizes not only may be extremely relevant for gathering data the column or in a separated study log document indicating
and generating knowledge but may also provide valuable the measurement units for each factor. Do not enter notes/
information to investigators interested in further testing comments in the same column as numbers or codes; make a
additional hypotheses. separate column specifically to add notes if needed. Clarity:
What about studies with large sample sizes? It is worth Give each column a short but logical title. If anything
noting that studies with large sample size have a greater needs more explanation, consider making a separate sheet or
precision and power; however, a large sample size in and of study log document with a key. This practice is valuable for
itself (e.g., 300 or even >1000 animals) does not guarantee making data easily interpretable to others, and for defining
validity and/or relevance just because of sample size. All more ambiguous variables (e.g., subjective scoring systems
key aspects of study design are also relevant to studies with or which treatment was assigned to which group) to reduce
large samples, and of particular interest in wildlife species, uncertainty when reviewing data in the future. Missing
potential confounding bias must be addressed in the study values: If any factor has missing information, leave the cell
design and data analysis, as well as proper interpretation of blank rather than adding not available (N/A) or unknown
the clinical, epidemiologic, or biological relevance of the (UNK), or adding a zero. If needed, make comments in the
obtained results (see Q10). comments column.
B
• Figure 5.2 (A) Suboptimal data structure/formatting for statistical analysis. (B) Optimal data structure/
• BOX 5.1 Rule of Thumb for Analysis to compare, number of factors being evaluated, and the
statistical test assumptions (including data distribution and
Proceed to the statistical analysis only after the data set has sample size) play an important role when selecting a statisti-
been checked for consistency, completeness, and accuracy. cal test. A standard process to select statistical tests includes
(1) specifying the hypothesis to test, (2) describing and
displaying the data graphically, and (3) checking the data
distribution and the statistical test assumptions. As a practi-
this scenario, it is strongly recommended not to ignore cal example, we chose a commonly used statistical test, the
it, which could result in overestimating the statistical Chi-squared test, to describe the approach to be used to
significance of results by artificially increasing the sample select a test. When the outcome of interest is categorical and
size. Some examples of statistical tests/methods that allow dichotomous, as often is the case when studying whether
accounting for lack of independence include the McNemar’s animals tested positive or negative to a test, whether animals
test, repeated measurements analysis of variance (ANOVA), are sick or healthy, and the investigator is only interested
and paired t-test,2–4 as well as the multivariable analysis1 in comparing two groups (e.g., females vs. males), then a
techniques summarized in Fig. 5.1. standard 2 × 2 table using the Chi-squared test to compare
the proportion of animals having or not having the outcome
of interest is appropriate. For this test, the assumptions are
Q9: Which Statistical Test/Method Do I that observations are independent and that the expected
Need to Analyze My Data? frequency in each cell of the 2 × 2 table is at least 5. If the
data do not meet these characteristics, then the Chi-squared
See Box 5.1. Most statistical and epidemiologic books test is invalid and a nonparametric option such as Fisher’s
contain self-explanatory flowcharts to guide investigators exact test should be used in scenarios with small sample
to select an appropriate statistical test.2,3 The selection size or McNemar’s test when data are not independent (e.g.,
of a statistical test/method largely depends on the type two screening tests used on the same blood sample). If the
of data of the outcome being measured (e.g., categorical investigator is interested in evaluating the impact of mul-
or continuous data). In addition, the number of groups tiple factors on the outcome (Box 5.2) (e.g., age, location,
26 SE C T I O N 1 General
• BOX 5.2 Multivariable Analysis from debate,15,16 the P-value is an important statistic to
consider but certainly not the only one. The P-value is the
The vast majority of outcomes studied in wildlife species are probability of making an error (type I error) in which the
determined by multiple factors. Collecting data on as many of
these factors as possible allows for a comprehensive statistical
investigator concludes that there is a significant difference
analysis. Multivariable models are techniques that include between groups (e.g., a drug reduces blood pressure) when
linear, logistic, and Poisson regressions, as well as survival in fact there is not. In general, P < .05 (<5% chance of
analysis models. These methods are the most commonly used error) is considered statistically significant. Having statisti-
analytic approaches to control for confounding by including cally significant results means that the observed differences
potential confounding factors in a model and thus evaluating the
combined effect of these factors on the outcome of interest.1
are not likely due to chance, and rather, differences in
the measured outcome are associated with the factor(s)
evaluated in the study. Important study design aspects,
such as sample size, power, the magnitude of the difference
being evaluated, and the amount of variability in the data,
time, and sex), then a multivariable approach using logistic have a direct and strong impact on P-values and should
regression analysis is recommended. Using this approach, be considered when interpreting results. Furthermore, it is
results may be adjusted for potential confounding effects, imperative to note that a study with a statistically significant
and, if needed, the investigator may control/adjust results result does not necessarily mean that the result is clinically
for lack of independence between observations to split the relevant, and vice versa. In studies with small sample size, it
variance between different aggregation (clustering) levels. is less likely to find statistically significant results; however,
In the scenario of a continuous outcome (e.g., blood findings still may have an important clinical, physiologic,
pressure in mm Hg, or calcium and phosphorus in ppm), or ecologic implication, especially if the magnitude of
the Student’s t-test (for comparing two groups), ANOVA the difference identified is biologically important. Hence,
(for two or more groups), and/or linear regression analysis although not replacing one for the other, the investigator is
(to evaluate multiple factors) are appropriate. When the encouraged to discuss both the statistical significance and
assumptions of these tests are not satisfied by the data, clinical importance of the obtained results.
nonparametric tests such as the Wilcoxon rank sum test,
Kruskal-Wallis test (nonparametric ANOVA), or transfor-
mation of outcome data to the logarithmic scale for linear Q11: How Do I Present and
regression are options to consider. Summarize My Results?
When the outcome of interest is counts (e.g., number
of diarrhea episodes in a period of time), Poisson regres- This largely depends on the forum in which the results
sion analysis provides a robust option because count data are being displayed, which may include scientific journals,
often (especially when the outcome is rare) have a Poisson or poster or oral presentations to a myriad of audiences,
distribution, which is characterized by the mean being equal including the scientific community, policy makers, or wild-
to the variance.1 When data are not independent, the main life management personnel. In general, when presenting
assumption of Poisson models (mean = variance) is often your results, after clearly outlining the goal of your study,
violated, and in these scenarios negative binomial models it is recommended to present a detailed descriptive analysis
may be used for analysis. In addition, if an excess of animals showing the distribution of both the outcome and factors
did not have the outcome (count = 0, no diarrhea) relative of interest. If the outcome or factor data are categorical
to those animals that did have the outcome, zero-inflated in nature, presenting proportions with their corresponding
models provide a robust option in which the researcher may 95% confidence intervals are recommended. If the data are
investigate factors impacting the probability of not having continuous and normally distributed, then means, standard
the outcome (0 counts) and factors impacting having a deviations, and 95% confidence intervals are appropriate to
greater number of outcomes among those animals in which summarize the data. If sample size is relatively small, use of
the outcome occurs. medians and range (minimum and maximum values), as
When the investigator is interested in the time taken for well as 25th and 75th percentiles, is recommended to sum-
an outcome to occur, survival analysis techniques includ- marize and describe the data distribution. When there are
ing life tables, Kaplan-Meier curves, and Cox proportional different groups of interest (e.g., treatment groups, species,
hazards models1,11–14 may be used, depending on the data sex, sampling points), the researcher should conduct a
distribution and study design. stratified analysis of any variables of interest for each group
of interest. If the investigator tested a hypothesis, then
initially univariable results describing associations between
Q10: Are My Results the outcome and only one factor of interest (e.g., changes
Statistically Significant? in lactate [outcome] values over time [factor] after immo-
bilization) should be presented. The limitations of descrip-
This is by far one of the most common questions among tive and univariable analyses, such as if factors known to
researchers. It is worth noting that, although not exempt impact the outcome were not considered in the analysis
CHAPTER 5 A Practical Guide for Statistics in Wildlife Studies 27
28
CHAPTER 6 Opportunities to Inspire the Next Generation of Veterinarians 29
time investment but have the potential to inspire many signage and prerecorded videos to explain the activities
new young people to pursue veterinary medicine. occurring. Others may have an educator explaining what
is happening in real-time. Veterinary staff participation
Veterinary Windows may be as minimal as simply performing the procedures
in the window, or could be as involved as speaking to
Some institutions have designed their veterinary hospitals the public through a microphone before, during, or after
to allow the public to observe veterinarians working in the procedure. Regardless of the level of involvement and
real-time (Fig. 6.2). These “windows” may be managed interpretation, these windows provide the visitors with a
in different ways. Some institutions may have scheduled glimpse into the high level of medical attention provided
procedures in the window on a regular basis and include to the animals in our care.
the time for the events in their visitor’s daily activities
schedule. Others choose to use the window for only special Public Dissections
events, such as education classes or tours. The degree of
interpretation also varies by institution. Some facilities use One interesting approach to educating zoo visitors about
animal anatomy occurs at several Scandinavian zoos includ-
ing the Copenhagen Zoo in Denmark. This institution
conducts regular “public dissections” of clinically healthy
animals that have been euthanized (Fig. 6.3). The dissec-
tions occur “behind the scenes,” but all visitors to the zoo
are welcome to attend. Veterinarians are able to discuss
comparative anatomy and display actual specimens at these
events in order to educate the public about the amazing
diversity of biology within the animal world. Feedback
from visitors has been overwhelmingly positive, as they
appreciate the opportunity to see anatomic features and
learn facts about animals that are hard to see or learn in
typical zoological programs (personal communication,
Mads Bertelsen, 2017) (see Chapter 23).
• Figure 6.1 Example of a tabletop display/booth at a special event
at the Central Park Zoo. Using prepared skulls, veterinary staff are Education Programming
discussing with visitors the variations in animal dental anatomy and
how that affects their diet and veterinary care. (Courtesy Julie Larsen Most zoological institutions have some form of an edu-
Maher © WCS.) cation department. These people are generally in charge
• Figure 6.2 Example of a specially designed window into the veterinary clinic to allow visitors to observe
live veterinary procedures at Disney’s Animal Kingdom. (Courtesy © Disney’s Animal Kingdom.)
30 SE C T I O N 1 General
TABLE
6.1 Sample Little Zoo Vets Class Outline
Opening: Introduction of instructors; scrub tops are distributed; photos are obtained to make name tag licenses for remaining
classes.
Didactic portion: Slide-based discussion
Physical exam: Whole body exam, use 4/5 senses (no taste), outline class for today.
Eyes: Using photos of normal and abnormal animal eyes, snake shed, and store-bought models, the following is covered:
eyelids, third eyelids, snake eye caps; cornea and lens, specifically the difference between cornea cloudiness and cataracts;
pupillary light reflex (PLR) using videos of a snow leopard with normal PLR, a macaw with voluntary pupillary movement,
and an owl with minimal response to light. Discussion of knowing what is normal for different species is important for zoo
veterinarians; eye position predator versus prey—view video of peacock walking after predator/prey activity.
Group activity: Predatory/Prey demonstration (field of view and depth perception). One student serves as the “predator” and
holds two purple strings in left hand and two green strings in right hand. With an eye patch over the left eye and while staring
straight ahead, the other students hold the ends of the purple string taut and move laterally until they are out of the right
eye field of vision. Repeat with the patch over the right eye. The field of view range is seen by the separation between the
lateral-most students, and the binocular/depth perception field is seen in the overlap between the two string colors. Repeat the
activity, this time with two students as the “prey,” one for each eye (so that they can be turned back-to-back to position each
eye laterally). Follow this up with tossing a ball to each student, first using both eyes open, then with an eye patch on one eye,
and compare the difficultly in catching the ball (depth perception). Proceed with discussion of how this knowledge might be
important to know as a zoo veterinarian.
Station activities: Small groups at each station will rotate for the remaining class period between: (1) practice using an
ophthalmoscope on a model eye (turn on/off, make the light blue, make the light a slit, look at the eye model, and draw what
you see); (2) palpation practice: using palpation models, determine what may be in the animal’s stomach using only your
hands, not your eyes (models consist of large animal prints with holes cut in the abdomen area, and prints are attached to
cardboard boxes just behind the holes and various objects are placed within the box)—students can then challenge each other
by placing new objects in the boxes for other students to try to guess; (3) body condition scoring: using printed scoring charts
and animal photos to give a body condition score to the various animals; and (4) live animal station: look at the outside of the
animal’s eye using the ophthalmoscope and describe what you see.
Materials list: Name tag supplies; photo release; class roster; camera; plastic eye models; sterilized snake shed; string and eye
patches; palpation cut-outs and box of “unknowns”; data sheets for ophthalmoscope practice, palpation, body condition
scoring; hand sanitizer; ophthalmoscopes; two live animals (ideally one mammal and one bird) with handlers.
• Figure 6.4 Example of the initial phase of most Little Zoo Vets • Figure 6.5 Little Zoo Vets student examining the eye of a guinea
classes where students are in a typical classroom lecture-style setting fowl using an ophthalmoscope at the Bronx Zoo. (Courtesy Julie
while the veterinary instructor is at the front of the room at the Bronx Larsen Maher © WCS.)
Zoo. (Courtesy Julie Larsen Maher © WCS.)
must practice their palpation skills to determine what those test and dissect a fresh elephant fecal bolus (Fig. 6.8). The
objects are. biomaterials used in the class are materials left over from
Some of the most enjoyable, though logistically difficult, regular clinical veterinary activities—none are collected for
parts of the class involve using real animal biomaterials. this purpose. Additionally, personal protective equipment
During the blood class, students are allowed to make blood is worn and students are monitored closely during these
smears, stain them, and examine them under the microscope. activities. A scrub top is supplied for each student and
During the feces class, students perform a fecal flotation laundered at the zoo between classes. Each student must
32 SE C T I O N 1 General
• Figure 6.6 Little Zoo Vets student scraping “tartar” (brown sugar)
from the teeth of a domestic dog skull at the Bronx Zoo. (Courtesy
Julie Larsen Maher © WCS.)
• Figure 6.8 Little Zoo Vets student dissecting an elephant fecal bolus
at the Bronx Zoo. Note the personal protective equipment being worn:
scrub top, eye protection, face mask, and exam gloves. (Courtesy
John Sykes © WCS.)
Conclusions Acknowledgments
Veterinary participation in education programs serve many Many thanks to the Veterinary and Education Departments
purposes, including increasing revenue, enhancing existing at WCS for their support of these programs and to all the
programming and the reputation of the institution, and students who have participated in our programs.
CHAPTER 6 Opportunities to Inspire the Next Generation of Veterinarians 33
References 3. Veterinary Medical Association of New York City: High School Vet-
erinary Career Exploration Program, 2016. Retrieved from http://
1. Wildlife Conservation Society: WCS.org: About Us, 2017. www.vmanyc.org/news-info/hs-vet-career-exploration-program/.
Retrieved from https://www.wcs.org/about-us. 4. Little Vets: Does your child want to be a veterinarian? 2017.
2. Wildlife Conservation Society: Educational Research & Evalua- Retrieved from http://www.littlevets.com/.
tion: Bridging the Gap, 2017. Retrieved from https://www.wcs.org/
education/educational-research-and-evaluation/bridging-the-gap.
7
Strategic Planning for Zoo
Veterinary Operations
SCOTT TERRELL
34
CHAPTER 7 Strategic Planning for Zoo Veterinary Operations 35
team members from all levels of your organization. As with scope of this chapter; however, there are numerous resources
all aspects of business and society, diversity adds strength available on the internet and in a variety of books on leader-
to an organization or a process. For a veterinary team, it ship, strategic planning, and business management.1,5–8
would be prudent to include key partners and stakeholders
(zoo operations, animal husbandry, education, etc.) in the Assess Your Overall Team/Environment
strategic planning process, as well as a diverse representation
of the skillsets/expertise on the veterinary team itself. Assessment of a team for the strategic planning process
may be accomplished via external or internal assessment.
Use of a Facilitator External assessments may come in the form of external
The use of a trained human resource (HR) facilitator or auditors, peer reviews, or accreditation inspections. Internal
outside consultant makes the strategic planning process assessments rely on the team/team members looking inward
easier but is not mandatory. One of the key roles of the facili- and honestly identifying strengths and weaknesses as part
tator (whether internal to the team or an external partner/ of the planning process. It is important to develop strategic
consultant) is to create a safe environment for exchange of objectives that build on team strengths and take advantage
ideas and free and open discussion, regardless of position. A of opportunities, while acknowledging and minimizing or
facilitator may also help to minimize introduced bias from overcoming weaknesses and threats. The simplest form of
the “boss” or senior leader in the room. Ideally, a facilitator a team assessment would come in the form of a discussion
should have experience with the strategic planning process with the members of the strategic planning team to identify
and some level of familiarity with the team or organization. basic strengths, opportunities, and external threats. Going
The facilitator is not the decision maker in the process but beyond a basic discussion, a number of other team/business
does ensure that all aspects of the process are addressed and assessment tools exist.
captured for the decision makers. One of the most common and most practical assess-
ment tools is the SWOT analysis, a tool that may help an
Set Aside Time and a Location organization or team identify internal and external factors
that contribute to the success or failure of strategic objec-
If you are going to plan, then plan. Plan to make time and tives.4,5 “SWOT” is an acronym for strengths, weaknesses,
plan to find a location as free from distractions as possible. opportunities, and threats. The “strengths and weaknesses”
This is easier said than done in the hectic world of a zoo vet- descriptors are often used to identify factors internal to
erinary practice; however, your team members must know the business or team, and the “opportunities and threats”
there is commitment to this process. That commitment will descriptors are used to identify factors external to the busi-
not be apparent in an environment of constant interruption ness or team.
or disruption. Depending on the size of your planning team The individual components of the SWOT analysis are
and the state of your existing plan (i.e., starting from scratch typically defined as:
vs. reviewing an existing vision and mission), it is reasonable • Strengths: characteristics of the business or team that
to expected that the strategic planning process will take at give it an advantage over others (internal factors).
least a half day of effort. When starting from scratch with • Weaknesses: characteristics of the business or team that
a new team or organization, that time commitment could place the business or team at a disadvantage compared
extend to multiple days. Prework in the form of questions or with others (internal factors).
discussion topics (see strengths, weaknesses, opportunities, • Opportunities: elements in the external environment or
and threats [SWOT] analysis later) may expedite the real industry/profession that the business or team could take
decision making and increase efficiency in the dedicated advantage of (external factors).
strategic planning sessions. • Threats: elements in the external environment or industry/
profession that could cause trouble or challenges for the
Create, Renew, or Review Your Vision and business or team (external factors).
Mission Statement The SWOT analysis process is driven by a set of ques-
tions designed to highlight each of the four categories.
Most organizations have some form of an existing vision Some examples of questions that could be used in a zoo
and mission statement. In those cases the strategic planning veterinary environment are provided in Box 7.3. Answers to
team will be focused on reviewing the existing vision and the questions may be captured in a 2 × 2 matrix such as that
mission or perhaps renewing/reenergizing the statements shown in Fig. 7.1. From this matrix, common themes are
themselves. If you are starting from scratch in an orga- identified to help identify strategic objectives and priorities.
nization that lacks a vision and mission, the first phase
of the strategic planning process is to identify these key Identify Strategic Objectives and Priorities
components. The key components of vision and mission
statements were covered earlier and several examples are SWOT analysis (or other similar assessment tools) may be
listed in Box 7.2. A detailed description of the visioning used effectively to build an organizational strategy. Steps
process and mission statement development is beyond the necessary to execute the strategy involve identification of
CHAPTER 7 Strategic Planning for Zoo Veterinary Operations 37
Weaknesses
• What is our team’s Achilles heel?
• Are there any institutional policies or practices that create
barriers for us?
• Figure 7.1 A typical 2 × 2 matrix used to format a SWOT analysis
• For what do our employees/team members criticize us the
upon which strategic objectives and priorities are set.
most?
• For what do our operating partners criticize us the most?
• What negative perceptions (true or untrue) do our employees/ In general, strategic objectives and priorities should be
team members/partners believe about us?
designed to leverage strengths of a team/organization and
• What elements of our business add little or no value?
• Is there a resource or process we lack that negatively take advantage of opportunities, while recognizing and
impacts our business? minimizing weaknesses and threats.
For example, a SWOT analysis of a veterinary/animal
Opportunities care team at Zoo X identifies veterinary expertise and
• Are there any new technologies, resources, or processes that hospital facilities as strengths in the SWOT analysis. At
we should be utilizing?
the same time a threat is identified from external “advocacy”
• Are there any new products, services, experiences, or
opportunities we should be offering? groups that seek to misrepresent or downplay the high level
• Are there any initiatives, external to our existing team/ of care that animals receive in zoological institutions. This
organization that could benefit us? threat has the potential to impact attendance and favor-
• Is there anything that our operating partners request of us ability among zoo patrons. Zoo X identifies animal care and
that we are not offering?
communication as strategic objectives over the long term
• As we look at other zoological institutions, what do they do
better than us? and develops strategic priorities for the upcoming year to
focus on continuous improvement of animal medical care
Threats as well as targeted communication of animal care–related
• If you were a zoo professional looking for a place to work, stories through social media and local media outlets. Special
would you choose us? Why or why not? goals may then be created for each of these priorities by
• Are there any initiatives, external to our team/organization,
a variety of team members. This is a generic example of
that could harm us?
• For what do our zoo/veterinary colleagues criticize us the the idea of building a strategic plan that capitalizes on the
most? strengths and opportunities of a team/organization while
• What negative perceptions (true or untrue) do our zoo minimizing weaknesses and addressing threats.
professional colleagues believe about us?
• Have the needs of the zoo community changed recently in a
way that we cannot react to? or influence? Communicate Your Strategic Plan
• What is/are the biggest threat(s) to the veterinary profession?
The zoological profession? A strategic plan is useless in the hands of only a few senior
leaders. To be truly effective, the strategic plan should be
communicated to all levels of an organization and to all
team members. As stated earlier, a well-designed and prop-
internal and external factors and selection of the most erly communicated strategic plan provides clear direction
important factors. From this analysis, one may determine for a team and may increase employee engagement and
the strategic objectives—those objectives so significant to effectiveness. The methods of communicating a strategic
the overall well-being of the team/organization that they plan are as varied as there are teams applying the process.
require defined efforts over time. The strategic plan should It is common for many organizations to use a one-page
focus on these objectives in combination with the overall format to highlight the basic components of their strategic
business environment to set specific strategic priorities. plan. One example of such a format is provided in Fig. 7.2.
38 SE C T I O N 1 General
Conclusion
A strategic plan may be one of the most important business
management tools available to a progressive leader, organi-
zation, or team. That same plan may also serve multiple
purposes to align work efforts, assist with priority goal
setting, motivate individuals, and establish accountability.
The components of a strategic plan may be defined with
• Figure 7.2 Template for concise capture of strategic planning key moderate effort through a process that a team of any size
components. may accomplish. In the end, the ultimate goal of any stra-
tegic plan for any team is to create an environment where
every team member understands the goals, priorities, long-
Numerous alternative formats exist and may be custom term objectives, mission, and vision for their organization.
tailored to the needs of your organization or team. In the context of a zoo veterinary team, the strategic plan
should align the veterinary team with the larger zoological
Empower and Expect Leaders and organization, ensure the veterinary team is providing long-
Team Members to Set Goals and term value for the organization, and provide a platform for
Establish Accountability the veterinary leadership to request support and resource
growth at the organizational level. A solid strategic plan
After your strategic plan is communicated to the team, it sets the framework for team and organizational success into
is time to start setting goals to accomplish the strategic the future.
priorities identified in the strategic plan. Goal setting is a
collaborative process between leaders and team members.
Identification of specific strategic priorities should make this References
process easier for both leaders and team members. If goals
1. Allison M, Kaye J: Strategic planning for nonprofit organizations,
are set appropriately (e.g., using the SMART model), then ed 2, Hoboken NJ, 2005, Wiley Publishing, Inc.
there should be a clear chain of accountability for specific 2. Bradford RW, Duncan JP: Simplified strategic planning, Worcester
goals and priorities. The goals and associated accountabil- MA, 2000, Chandler House Press.
ity may then be used for periodic or annual performance 3. Davis JR, Frechette HM, Jr, Boswell EH: Strategic speed, Boston
reviews, performance management, or recognition/reward. MA, 2010, Harvard Business Press.
4. Gregory A: How to conduct a SWOT analysis for your small
Review in Regular Intervals and business. https://www.thebalance.com/swot-analysis-for-small
-business-2951706. (Retrieved 27 April 2017).
Amend as Necessary 5. Olsen E: Strategic planning kit for dummies, ed 2, Hoboken NJ,
To ensure the strategic plan performs as designed, you must 2012, Wiley Publishing, Inc.
6. Scott CD, Jaffe DT, Tobe GR: Organizational vision, values, and
hold regularly scheduled formal reviews of the plan and
mission: building the organization of tomorrow, ed 2, Menlo Park
refine as necessary. Ideally, a schedule should be established CA, 2011, Crisp Learning.
to review the progress of strategic priorities at least on 7. How to Write Your Mission Statement. https://www.entrepreneur
a quarterly basis, if not more frequently. Establish clear .com/article/65230. Entrepreneur. (Retrieved 1 February 2017).
accountability for progress or completion, but be willing 8. How to Write a Powerful Mission Statement. https://www
to amend goals and priorities in a dynamic environment. .kinesisinc.com/how-to-write-a-powerful-mission-statement/,
It is ridiculous to think that components of the strategic www.kinesisinc.com. (Retrieved 1 February 2017).
8
Organizational Influence:
Navigating the Leadership
Road for Zoo Veterinarians
DONALD L. JANSSEN
39
40 SE C T I O N 1 General
the person, the team, the organization, and the whole zoo veterinarian may think his or her role is to speak for the
profession. animals and to represent their departments at the executive
Traps and obstacles to becoming influential abound. level. The executives welcome those who can step away from
If veterinarians learn to overcome these barriers, though, their singular focus as veterinarians and see the big picture.
they can make the most of their value and serve their It takes humility to change focus, listen and learn, and serve
organization to a greater degree. If they have influence, they the whole organization. Those who do will be in a strong
are less inclined to suffer burnout and more likely to have position of influence, which, coincidentally, will benefit the
a satisfying career. Studies looking at physician burnout animal focus as well.
show that having influence and meaning at work are drivers Humility is a leadership attribute that one may develop.
for engagement and prevent burnout.1 This chapter is a Several character-based behaviors describe humble leaders.
compilation of principles and practices designed to help For example, admitting mistakes, managing emotions,
overcome the obstacles. The ideas come from a study of the being honest, being an effective listener, delegating decision
leadership literature and my journey, mistakes and all, in making, and giving recognition are among the behaviors
leading zoo animal health teams. Further insight has come that depict humble leaders.2 With motivation and practice,
from interviews with successful zoo leaders. This chapter is anyone can be an authentic and humble leader. A humble
not a formula for success. My results have been far from leader puts others at ease, which in turn, produces lasting
the ideal. Rather, I offer this as one path to follow toward relationships with those they need to influence.
becoming a more influential leader.
Seek to Serve Others First
People: How Relationships Impact Having humility leads to other attributes that help us grow
Organizational Influence our influence. One such attribute is a willingness to serve
others for the greater good.4 Most people object to the idea
The quality of work relationships has a direct impact on that their job is to serve others. Still, having an attitude
organizational influence. Amid the busyness and scope of of service is key to developing organizational influence.
a zoo veterinarian’s job, managing the care of each animal Approaching the situation with an attitude of service shows
takes priority. However, to gain influence and make a that the intention is to put others’ needs ahead of one’s own.
long-term impact, nurturing the many work relationships In other words, a leader’s job is to serve, not to be served.
is crucial. Being a zoo leader means being good at relation- Good leaders set direction, then turn the organizational
ship building. The following sections discuss six building chart upside down. These leaders, in essence, work for their
blocks that zoo veterinarians may use to form healthy people. They use their position to remove obstacles for
relationships. getting work done. They put the focus on others and not
on themselves.5
Begin With Humility When putting servant leadership into practice, it is
helpful to ask clarifying questions to help direct one’s
Successful relationships start with genuine humility as their behavior: Whom do I serve? Is my focus on myself or
foundation. Humility connects people through a common others? How may I help the situation? When things go well,
human bond.2 A humble person has a modest and accurate who gets the credit? Am I willing to be vulnerable? When
assessment of his or her importance and abilities. Humility and how do I say “no”? Do those I lead grow as people? Do
requires a high degree of self-awareness and empathy. Being they, while being served, become more autonomous, more
humble is not a sign of weakness; indeed, it requires a good likely themselves to become servants?4 The answers to these
deal of strength and assertiveness. Humble leaders assert questions help us distinguish an outward (selfless) mindset
themselves on behalf of their team’s accomplishments rather from an inward (selfish) mindset.6 The practice of servant
than for themselves.2 Humble leaders have self-confidence leadership has the power to transform both the leader and
but do not project a feeling of self-importance. Humility is those who are served so that respectful relationships may
power under control. flourish.
We trust and enjoy following leaders who are humble
and not self-serving. Humble leaders are effective and influ- Put Relationships Above Results
ential. Jim Collins labels the most successful leaders as Level
5 leaders. He describes these leaders as having both humility Conventional wisdom says we should direct our best efforts
and fierce resolve.3 Zoo veterinarians, in general, have the at achieving goals and results. Good animal health and
resolve. Lacking humility, this resolve can be intimidating welfare outcomes are, indeed, the results we are after. Vet-
and objectionable. Veterinarians can be intimidating just erinarians must also pay attention to business and financial
because of their position and education. Those who act in goals. To achieve these desired outcomes, it requires teams
a humble manner can counter this tendency and be the of people working together effectively. Teams function best
welcome exception. Working effectively at the executive when they make trusting relationships their foundation and
level is an example of where humility may be an asset. A priority.7
CHAPTER 8 Organizational Influence: Navigating the Leadership Road for Zoo Veterinarians 41
Great leaders can be the model for developing trusting contributing to the process, that veterinarian demonstrates
relationships within their teams. To illustrate, imagine an integrity (character) and capability (competence). Making
animal health crisis, such as a sudden medical emergency in a commitment creates hope. Trusting relationships arise out
an elephant. The organization’s leadership is often present of this practice.
at the scene. However, the purpose should not be to direct Trusting relationships are essential in accomplishing the
or judge the actions of the animal care teams. Instead, critical work of zoos and aquariums. We depend on many
leadership’s role is to be present and care for the needs of key stakeholder relationships for animal care. Be smart
the people. The attention by the leadership ensures that the about trusting others but be willing to do so. “Be” (charac-
animal care team can achieve the best result possible. In one ter) and “do” (competence) in ways to increase trust with
institution, this philosophy has played out in many crisis others. No relationship is perfect, and everyone betrays trust
situations and has become an unspoken rule. “In a crisis, occasionally. But strive for high-trust relationships. They
pay attention to the people first.” The principle is just as are the foundation for developing organizational influence.
important in daily work as it is during a crisis. Pay attention
to relationships first and collective goals and results will Honor Your Staff
follow. Putting people first bolsters team trust, influence,
and ultimately the health of the whole organization. For obvious reasons, we aim to influence the high-level
decision makers in our organizations. However, in doing so,
Build Trust we may forget about the importance of our staff. More than
most, the actions of our teams are critical to our success in
Diversity of abilities and unity of purpose are the twin establishing organizational influence. An excellent support
engines that power success in teams. If diversity is the source staff understands that their leaders care about their well-
of talent on a team, then trust is the glue that holds the being. These leaders honor their team as individuals and as
team together and creates unity. By serving others with a group. Honor is a status we give to others. It builds them
humility, we depend on trust to protect and sustain the up, raises their dignity, and prepares them for a higher level
relationships. We need the trust of our coworkers to have of performance. Honoring the staff means treating them
successful outcomes for the animals under our care. Each of as if it were your job to serve them rather than vice versa.
us can think of a person with whom we have a high-trust It is unconditional and separate from accountability and
relationship. Those relationships feel special. Communica- performance (which you still must address).
tion is quick and easy. Things get done with little effort. It is Honoring those with less authority fosters loyalty and
enjoyable. The opposite is true with a low-trust relationship. wards off unhealthy conflict. Employee satisfaction surveys
High-trust relationships produce exceptional financial and usually include opportunities to provide comments. The
mission-related results.8 most troubling comments come from those who report
Trust engenders confidence, and the lack of trust leads being treated harshly by those in authority. People want
to suspicion. There is a continuum between confidence and to be treated with dignity and respect and not valued just
suspicion. But a “line of trust” exists between these two for a function they serve. Veterinarians, because they are
states and yields two distinct outcomes. Above the line, viewed as authorities, are in a unique position to turn that
communication flows, and assumptions about each other’s around and bolster the status of others. This recognition
behavior are good. When conflict arises, we have faith that will give them a reason to be loyal and engage in the greater
we can resolve it. Below the line nothing is easy, and doubt vision. One way to do this is to give particular credit to staff
prevails. Everything becomes an issue. We lose confidence members for team achievements instead of implying credit
that we can work out a conflict. We slip above and below for oneself. As an example, one leader that I know has a
the line based on our choices and actions. However, we can reputation for honoring his close staff. In a variety of ways,
build up “relationship equity” over time. In doing so, our he makes sure they know that their personal well-being is
position relative to the “line of trust” becomes more stable. important to him. He advises his teams to do the same
Little transgressions then have a lesser effect on the position by saying “It’s how we do things here. It’s the people that
of our line of trust. matter.” Honoring your staff is one of the most valuable and
One can understand how to build trust by viewing it as underutilized tools for developing organizational influence.
a function of character and competence. In the veterinary
context, character is similar to professionalism. Competence Connect With Purpose
is related to technical ability.9 Character includes our integ-
rity, motives, and intent with people. Our character is who When teams feel appreciated, they become more motivated
we are. Competence includes our capabilities, skills, results, and may become tremendously influential. Motivating
and track record. Our competence is what we do. We can people is a leader’s number one job. Strict accountability is
use this concept in a practical way to build trust. Building one approach used to motivate. Rewards and punishment
trust requires that we make and keep commitments.8 For do have a place in managing people. But good leaders
example, a veterinarian may commit to helping design do not rely on them as the primary tools for motivation.
a new exhibit. By honoring the team commitment and These tools create a transactional environment that may
42 SE C T I O N 1 General
feel like manipulation. Employees lose their desire to be The good news is that we can overcome our desire for
self-motivated, requiring even closer supervision. control by adhering to an important principle. Simply
Instead, seek to inspire with an emphasis on “why.” In stated, it is this: to gain influence, give up control. That
other words, work to discover the team’s purpose and be is, to obtain long-lasting influence, abandon the desire for
good at telling the “why” story. Pay attention to their hearts, authority and control. Yes, this principle is counterintuitive,
that is, the intrinsic motivation. Understand their deepest so most of us would rather disregard it. We often hear that
drivers and relate that to a common purpose, the “why.”10 striving for control is the noble thing to do and the way
When people know and believe in the purpose, they moti- to ensure the right decision is made. But, we work with
vate themselves and one another. Accountability becomes people. So, if we wrestle with people for control, we will
less of an issue. Our profession and what we do lends lose valuable influence, trust, and credibility in the long
itself well to this approach. What we do has a compelling run. Instead, we may choose to respond with humility and
purpose and is motivating in itself. The role of the leader respect, treating others as people with needs and desires
is to remind people of the overarching purpose and how it similar to our own. We can act to serve their needs as well
relates to the job at hand. as our own. If we follow this path, our influence in the
Besides the constant reminders of the high-level vision, organization and with other people will soar. Application of
one must also be ready to share the “why” of the moment. this principle may lead to strong, trusting partnerships with
Veterinarians going into procedures are good at telling the synergistic outcomes. In the end, giving up control leads to
“what” and the “how.” They are good at telling staff what more influence and better decisions.
they are going to do and what they want them to do. Further practical steps flow from this principle. Once
They are also used to telling people how they are going a partnership develops, the parties clarify their roles. One
to do something and how they want them to assist. But party takes on the “Decider” role and the other the “Adviser”
how often do we take the time to explain the “why”? The role for a given decision or type of decision. The Decider is
“why” gives context and meaning to a procedure or activity. the responsible party and has the ultimate decision-making
Sharing the “why” assures those working with us that we authority. Good Deciders seek input from those in the
have thought through the procedure. It makes it clear that Adviser role; they take full responsibility for outcomes and
what we are doing is important, necessary, and part of a blame no one if things go wrong. In contrast, the role of the
greater purpose. The “why” connects the vision to the role Adviser is to influence the Decider. They may change the
of animal health and to the task at hand. It reminds people minds of others by providing evidence, interpretation, and
that together we are making a difference. Sharing the “why” recommendations. Their manner must be professional and
of the moment is another way of honoring and inspiring respectful, honoring the Decider’s position. Veterinarians
our staff. An inspired staff is a productive staff, and that often find themselves in this Adviser role. They have special-
enhances influence at all levels. ized knowledge and often uncover the problem first and in
the most depth. So, they may be the primary drivers in the
decision-making process by initiating dialog and providing
Practical Steps: Taking Your Influence their perspective. It is important to remember that neither
to the Next Level role is more valuable than the other, and both are necessary
for making good decisions. Indeed, successful partnerships
Once we appreciate the significance of strong relationships, are not based on equal and identical functions. Rather they
we are in a good position to adopt specific practices to grow thrive when roles are defined and distinct.11
our influence and that of our teams. Good practices should Giving up control to gain influence is a paradox. Even
build upon relationships by clarifying roles and expectations so, it is a powerful servant leadership principle. It is useful
for conduct. They should encourage leadership growth in for those without positional authority who want to have
others who can, in turn, become influencers. The following influence. Giving up the desire for control is a principle
sections provide a few such practices to consider for increas- worth understanding and applying in our work and home
ing organizational influence. environments. It achieves better results and becomes the
right thing to do in serving others.
Give Up Control to Gain Influence
Create Principle-Based Standards
Most of us are eager to influence decisions that could affect
us, but trouble may arise when those involved in deci- Having accountability for ourselves and in the teams that we
sion making have conflicting perspectives. We all want the lead shows responsibility. Accountability gives us credibility
animals to thrive under our care. Why, then, would conflict with others and builds our global influence. But holding
occur when making decisions about their care? The answer ourselves and others responsible is tricky business. Caring
lies in our human nature. Human beings have a desire more about rules than people invariably leads to opposition
for authority and control. This desire works against our to those standards. But how can we keep people accountable
ability to influence others, especially when controversial or and still serve our staff’s needs? How can we uphold high
high-stakes decisions come up. standards without damaging relationships? One approach
CHAPTER 8 Organizational Influence: Navigating the Leadership Road for Zoo Veterinarians 43
is to create principle-based standards. Accountability, then, diminishes the opportunity for others to develop their
becomes part of the culture. In addition, having the team leadership skills. Even more, it leads to confusion around
develop the standards themselves helps make them relevant daily tasks and assignments. Workers abdicate responsibility
and accepted. Without such standards, each person tends and avoid making decisions. Errors become more prevalent.
to do what is right in their own eyes, leading to confusion In these environments, it is characteristic for employees to
and conflict. use language that shows their dependence and insecurity.
Working together in our animal health departments, Before taking action, they may use weak phrases, such as:
we designed seven principle-based behavior standards to “I would like to…” “What should I do about…” “ I have
ensure accountability. We wrote them to be consistent with no authority to…” “It is not my job.” “Tell me what you
our broader organization’s code of conduct.12 We put the want me to do.”
phrases into our words so they would make sense in our work A simple practice of using empowering language may
environment. For each standard, we wrote a catchphrase to reduce this dependent behavior. The object is to push deci-
make the concept more relevant. We also wrote clarifying sion making to the level where the information is. This
questions to help identify specific, desirable behaviors. method empowers people and develops them as leaders.
Standards should reflect the unique nature and values It transforms insecurities into confidence. It builds trust
of each workplace to be most effective. Here is an example and reduces errors. Empowering phrases, encouraged and
of the format and wording for one of our standards. Code modeled by those in authority, show a leadership intent,
of Conduct: “Use Your Words Wisely.” Our rewording: and that trust is implicit. Here are some example phrases
“Think before you speak.” Catchphrase: “My words and that signal confidence when taking action: “I intend to…”
nonverbal messages affect those around me and our work “I plan on…” “I will…”13
environment.” Clarifying questions: Am I respectful? Am Using this approach to empower may play out in a variety
I phrasing my message in a positive manner and offering of situations. The following is an outline of the process and
solutions? Am I addressing the issue directly with the person gives examples showing how this may work. (1) Any staff
involved? Do I express gratitude whenever possible? member, regardless of his or her position in the department,
The remaining six standards along with their catchphrases sees a need for action and knows what to do. (2) That staff
were: Take responsibility (By holding myself accountable for member tells the responsible party (e.g., his or her boss,
my actions, there is no one else to blame), work effectively veterinarian, peers) that he or she intends to take action.
(When I use my time wisely, the entire workplace benefits), (3) The responsible party acknowledges the intention. He or
build excellence (Bring out the best in yourself and others), she may ask clarifying questions and agree to the action or
harmonize (I can achieve harmony when I balance my make a change as needed. The roles are crystal clear. Author-
work and personal life), create joy in the workplace to ity and responsibility are in their proper context. Feelings of
relieve stress (I can help create a pleasant and respectful being micromanaged vanish. Delegation is effortless. This
workplace), and make lasting memories (Create a legacy of process pushes the authority to act down to the source of
learning, innovation, service, and leadership). Again, these information. It eliminates the confusion of who is going to
are examples based on a particular work environment and decide to act; it reduces time wasted waiting for someone
would necessarily be different elsewhere. to order an action; it avoids mistakes; individuals assert
With agreed-upon, principle-based standards, a common themselves; they make full advantage of their knowledge
language emerges for acceptable behavior. A culture of shared and experience but get the opportunity for others to check
accountability results. To sustain these practices, then, the their reasoning; it enables employees to grow in their jobs;
leader’s job is to provide a venue for repetition and remind- and they become leaders themselves.
ers. Some workgroups choose to review a standard at the
beginning of each day. A practice such as this gives everyone Example 1
an opportunity to apply and reinforce the behaviors and
develop leadership skills as well. Situation: A manager noticed that an area in the hospital
needed urgent repair. Stated intention: The manager got
Develop Leaders a bid for repair. She reported to her supervisor that she
intended to institute the repair during the next budget
If we want sustained organizational influence, we need to period. She was confident that she would stay within year-
invest in others and help those around us become leaders end budget, but expenses would be over for the current
themselves. Developing leaders is not just for succession period. Intention authorized: The supervisor paraphrased
planning. It is essential for smooth daily operations and the issue to make sure she understood it. She asked ques-
successful outcomes. In such an environment, each worker tions about the urgency of the repair and then agreed to the
becomes a leader. Each employee, regardless of rank, takes action. Result: The action happened without delay by the
responsibility and has the authority for his or her area of person with the most information to take decisive action.
expertise and duty. Often, though, as the authority figure, Other stakeholders were informed and had an opportunity
we have the need to appear in charge, give directions, and to weigh in on the decision. The manager took personal
be the source of knowledge. This authoritative approach responsibility. She checked her reasoning with others, which
44 SE C T I O N 1 General
Introduction See Table 9.1 for a quick reference list for acronyms used
in this chapter.
Anyone who is involved in the management of non-native
or indigenous wildlife understands that there are inherent
risks when working with these animals. These include risks What Is Contingency Planning?
from the animals themselves, and external risks such as And Why Is It Needed?
natural disasters. Incidents such as “big cat” escapes, high-
impact flooding events, and infectious disease exposure Contingency planning is the development of plans and
may severely impact a facility, its staff, its visitors, and procedures that may be implemented when normal operat-
its animals. ing procedures are disrupted due to incidents or events.
Following an incident, there is typically increased scru- Adverse media attention is certainly not the only reason
tiny of the facility, and questions arise about the staff’s that contingency planning should be a priority in animal
ability to manage collections in the event of a future disaster. management. For example, it is very important to have a
In a 24-hour news cycle, the media is able to seize upon a contingency plan for dangerous animal escapes or unin-
story and, aided by social media, repeat it again and again tended contact situations between the animals and the
until it soon “goes viral.” Consider the 2016 shooting of a public or staff. It is paramount to avoid injury or loss
great ape in response to a breach of an exhibit. This incident of life of any persons or animals in such situations. For
was a topic of conversation for months, and a cell phone facilities that house endangered species, lack of planning
recording of the event has since been viewed millions of for an incident could mean a significant loss to the species
times. The incident resulted in protests, petitions, memes, gene pool. The health and welfare of these animals during
and news stories. Despite a sound explanation that this an incident should motivate facilities to begin the planning
action was taken to save the life of a child, it still proved to process and bring it to a purposeful status. As a bonus,
be extremely challenging to manage the perception of the planning may also increase the efficiency of response and
public, versus the expertise of the animal’s keepers. This the safety for responders.
event highlights the increasing public scrutiny on facilities
that manage these animals, and their ability to plan for Contingency Planning Principles
emergencies.
There are a wide variety of facilities that own, breed, If a facility has yet to begin development of contingency
and/or exhibit non-native or indigenous wildlife species as plans, whether for a natural disaster, animal escape, or foreign
their mission, their hobby, or their business. This includes animal disease, there are planning principles recommended
accredited and nonaccredited zoological collections and by the Federal Emergency Management Agency (FEMA)
aquariums, drive-through animal parks, exotic game ranches, Course IS230c: Fundamentals of Emergency Management1
animal sanctuaries, and private individual owners and/or that provide a strong foundation. Adapted for exotic animal
breeders of these species. There may be multiple authorities and other wildlife facilities, these principles are listed below.
and statutes that govern their activities, depending on the • Planning must be community based, representing the
type of species cared for, the state in which the facility is whole population and its needs. A facility should rec-
located, and the intended use of the animals. Each indi- ognize that they are part of the larger, interconnected
vidual facility must know which statutes, laws, or agencies community.
have jurisdictional authority with regard to the species in • Planning must include the participation of various stake-
their care. holders and subject matter experts in the community.
45
46 SE C T I O N 1 General
TABLE
9.1 Acronym List
Acronym Definition
CAP Control Access Point: Designated sites where various levels of biosecurity and protocols change to protect
collection and or staff
CPG Comprehensive Preparedness Guide: Documents developed by Federal Emergency Management Agency to assist
with contingency planning
EM Emergency management: A professional discipline that prepares, prevents, mitigates, responds to, and assists in
recovery from disasters
FAD Foreign animal diseases: Diseases not normally found in the United States. Detection triggers intense response
action and often results in trade restrictions on agricultural commodities
FADPReP Guidance developed by US regulatory officials that outlines national strategies for the control of foreign animal diseases
FMD Foot-and-mouth disease
ICS Incident Command System: A structure used by animal health regulatory officials to organize a disease response
LOS Line of Separation: A structural or functional division between areas for defining varying levels of biosecurity
NAHEMS National Animal Health Emergency Management System: Series of guidance documents for the management of
infectious diseases
SAHO State Animal Health Officials: Also known as State Veterinarians in the United States
THIRA Threat and Hazard Identification and Risk Assessment: Standardized process used to assess the likelihood of incidents
USDA US Department of Agriculture: One of its many missions is the prevention of and the response to foreign animal
diseases
Facilities may find value in joining or participating in respond to many incidents. Local EM efforts are maximized
local response planning groups led by Local Emergency when businesses within the community participate in the
Management. planning and capability building process. EM professionals
• There are proven processes for the development of may provide knowledge of future plans to mitigate the
contingency plans. impact of a particular hazard; for example, a new levee
• Planning starts by considering all threats and hazards. planned in the next 3 years to minimize flooding. They have
• Plans should be flexible and scalable. This means that knowledge of local resources and may serve as a conduit to
planning considers, for example, one antelope escape them if needed in a response.
but can expand for the escape of a dozen. Optimal planning team composition will vary depending
• Plans should clearly identify goals and objectives. on many factors, such as size, location, species, and site-
• Planning does not need to start from scratch. specific risks. Ask other facilities with robust plans about
• Planning identifies tactics and tasks needed to fulfill their planning team composition, and the value that diverse
objectives. backgrounds added to their planning process. A veterinary
• Effective plans outline what to do and why to do it. professional should be on the team. This involvement is
Roles and responsibilities are outlined. imperative for a number of reasons; the facility veterinarian
A fundamental truth in planning is that the planning may educate EM professionals about the collection and
process is more important than the written plan itself. The specific risks. They will be critical in explaining the capa-
relationships and partnerships forged with the local response bilities of the facility’s animal health paraprofessionals, and
community during the planning process may determine the will serve as the conduit to State Animal Health Officials
success or failure of plans during a catastrophic event. (SAHOs) and other regulatory personnel when a foreign
animal disease (FAD) or zoonotic disease is suspected.
Facility managers will be key participants, providing an
Contingency Planning Step 1: understanding of the facility footprint and infrastructure;
Form a Collaborative Team additional planning team members may include keepers
and curators.
Regardless of whether your facility has been included in local As a part of the planning process, first responders must
planning efforts, it is the responsibility of facility owners or be included; local police, fire departments, and emergency
operators to include community emergency management medical services should be involved in plan development
(EM) officials on your plan development team. Without or review. SAHOs must also be included or consulted,
this integration, you may NOT be adequately prepared to especially when planning for an FAD event. Their expertise
CHAPTER 9 Contingency Planning for All Hazards and Foreign Animal Disease 47
will be needed to explain the roles and responsibilities of Foot-and-mouth disease (FMD) is a threat that Federal and
the facility, the State Animal Health agency and the US State animal health professionals have invested significant
Department of Agriculture (USDA) in FAD outbreaks. It resources in prevention, mitigation, response, and recovery
is wise to discuss the resources (trained responders and planning efforts. FMD has been used as a disease model
equipment) available at the state level and facility. If because of its highly contagious nature to multiple species
your state government has an Animal Health Emergency and the agricultural trade implications of detection. To begin
Management coordinator, you may find that to be a good the risk assessment thought process for FMD, consider the
contact as well. following equation:
As the team begins its work, it is highly recommended
Hazard + Likelihood + Vulnerability + Consequence = Risk
to discuss the use of the Incident Command System (ICS).
ICS is an integrated, organizational tool that is flexible and Discuss the following with the team during the risk assess-
scalable and has been in use by EM, fire fighters, and State- ment process: (1) Do you know what FMD is? (Hazard).
USDA disease Incident Management Teams as a framework (2) Is the disease common or likely to emerge? (Likelihood).
to organize responses to incidents. It is recommended that (3) Does the facility have animals that can contract FMD or
development of your plans consider the use of the ICS serve as amplifiers or reservoirs of disease? (Vulnerability).
framework in order for the facility to fully integrate into (4) How would the detection of FMD in the facility or
the coordinated response. Basic ICS training is free and community impact business operations at the facility, state
available on the FEMA Emergency Management Institute’s agriculture, or national agricultural levels? (Consequence).
website.2 Training may also be available directly through Upon completion of a thorough risk assessment, it is recom-
your local responders. mended to begin planning for “high likelihood and/or high
The facility planning team should consider appointing a consequence” events. FMD may be a low likelihood event,
leader who may assign tasks, develop deadlines, and engage but the risk assessment process should include this disease if
the assistance of other subject matter experts whose input the facility manages susceptible species, as the consequence
may be needed as plans are developed. likely would be devastating.
biosecurity possible to avoid infection to its own animals plan approval. Other elements of this section may include
as well as to any other susceptible animals nearby. It is planning assumptions, such as recognition of the SAHO’s
important to remember that this responsibility will be authority in the management of FAD events. Supporting
borne by the facility, to the level acceptable by SAHO annexes cover topics organized by agency or function. For
and USDA. example, a facility may wish to develop an Emergency Elec-
• Animal Movement: Non-native or indigenous wildlife trical Outage Annex after having identified that damage to
move daily in the United States, but the numbers are the electrical grid or supply lines could be caused by any
miniscule compared to the staggering number of agri- number of hazards identified in the risk assessment (ice
cultural livestock moved intra- and interstate. Livestock storms, tornadoes, rolling blackouts). Additional support-
move according to the ages and use of the animals, so it ing annexes may be considered for emergency communica-
is not surprising that the urgency to move non-native or tion, shelter-in-place plans for people and animals, and
indigenous wildlife in the short term is also dependent evacuation plans for people and animals. Disease response
on the facility’s operating model. Zoological parks and plans are ideally suited for a hazard- or incident-specific
sanctuaries may seldom move animals in their collection, annex because of the unique circumstances posed by a FAD.
and as such, may not initially feel impacts from restric-
tions on movements of animals and animal products
during FAD outbreaks the way a game ranch relying on Development of Foot-and-Mouth Disease
movement of hides, trophy mounts, and other hunting Incident Annex Using Secure Zoo Strategy
products might. However, if the restrictions last long
enough, movements required for species conservation The development of an FMD annex to a facility plan will
and breeding, to fulfill agreements between facilities, or be challenging, but there is help available to assist with
to meet dietary requirements of certain animals in the this process. Agricultural industry groups have developed
collection could be affected and potentially become a tools known as “Secure” plans to support business continu-
financial disaster for facilities with that business model. ity for the industry as a whole, as well as for individual
• Visitation: In facilities that rely on some form of facilities/farms. Plans currently under development include
visitation, the restriction of paying visitors to view or Secure Poultry, Secure Milk, Secure Beef, and Secure
hunt animals in an effort to prevent disease spread Pork.6 Each plan discusses specific details, challenges, and
could financially devastate the facility (as happened in concerns about preparedness, response, and most notably,
the United Kingdom during an FMD outbreak). If a recovery from catastrophic disease events. In an effort to
contagious FAD outbreak occurs near a facility, visitation provide similar “Secure” tools to facilities with exotic or
could be temporarily restricted. If a goal is to resume indigenous wildlife, a Secure Zoo Strategy project has been
visitation as soon as possible, plans must be written with launched.
that goal in mind. Secure Zoo Strategy7 development is currently under
As plans for FMD are being developed, facilities should the guidance of the ZAHP Fusion Center. It should be
include quantifiable goals whenever possible; with pres- noted that Secure Zoo Strategy material is not just for
ervation in mind, a quantifiable goal may be: “The goal facilities that identify themselves as zoos. There is broad
for Acme Zoological Park is to prevent FMD infection applicability to anyone managing FMD susceptible species.
in their Nigerian Giraffe.” This goal will require develop- Subject matter experts, including SAHOs, virologists,
ment of biosecurity objectives to protect the endangered epidemiologists, and representatives from agriculture and
giraffe. These objectives will drive discussion of strategies other sectors have contributed to the development of Secure
and tactics that will become part of the written plan. To Zoo tools. Like the other “Secure” plans, guidance comes
ensure the greatest possibility of success, every goal included from FAD Preparedness and Response Plans (FADPReP).8
in the plan should have buy-in from the owners, opera- These planning documents along with the Foot-and-Mouth
tors, appropriate governing officials, and staff who will be Disease Response Plan: The Red Book9 (draft September 2014)
implementing the plan. are available on the USDA FADPReP website. Secure Zoo
Strategy (like other Secure plans) is a work-in-progress;
however, these tools are designed to help facilities working
Contingency Planning Step 4: with their SAHO to draft their own FMD hazard-specific
Plan Development disease annex. Facility owners and operators must recognize
that the responsibility lies with them to protect their animals
This phase of planning is critical in identifying what ele- as much as possible.
ments should be included in your all-hazards plan. Consider Secure Zoo Strategy proposes a series of steps to guide
the following three plan components: (1) the basic plan, a facility through the planning process for prevention and
(2) any supporting annexes, and (3) hazard-specific annexes response to an outbreak of FMD. Secure Zoo Strategy
(such as an FMD annex).1 A basic plan should include intro- encourages the planning team to understand the goals,
ductory material validating the plan and process. This may objectives, and terms discussed in the current FMD Red
include the plan, purpose, and scope, and the signatures for Book to ensure the facility, SAHOs, and USDA are all using
CHAPTER 9 Contingency Planning for All Hazards and Foreign Animal Disease 49
TABLE
9.2 Premises Designations*
*This summarizes the premises designations that would be used in an FMD outbreak response.
FMD, Foot-and-mouth disease.
Reproduced from United States Department of Agriculture Animal and Plant Health Inspection Service. FADPReP Foot-and-Mouth Disease Response Plan:
The Red Book. Available at: https://www.aphis.usda.gov/animal_health/emergency_management/downloads/fmd_responseplan.pdf. Accessed January 2017.
consistent terminology in the planning effort. Obtaining implemented for numerous disease concerns. The develop-
a copy of the State’s animal disease incident annex (if ment of a sound program is important, but the ability
available) may provide valuable insight into the goals and to consistently implement it is critical. A Biosecurity
objectives of the State’s plan. How robust the State Animal Program will need to identify both Structural Biosecurity
Response planning efforts are will likely be reflected by elements, such as fences, walls, or other physical barriers
the size of the food animal industries located within that that prevent possible contact with diseased animals, as
state. One of the first steps of the strategy is to adopt and well as Operational Biosecurity elements, which consist of
understand how Premises Designations (Table 9.2) will be personnel protocols, equipment use and disinfection, work
used to coordinate individual facility response. flow considerations, and other protocols to prevent the
The planning team should discuss the implications of spread of disease. Using the concept of increasing Levels of
each Premises Designation that may apply to their facility Biosecurity, a facility may identify places in the plan where
if an outbreak were to occur nearby. These implications specified events would act as “triggers” indicating the need
will include additional responsibilities, such as increased to increase biosecurity activities, thus moving the response
biosecurity and surveillance, and restrictions to animal to the next level. Secure Zoo Strategy recommends this
movement and visitation. Discuss this in context with the concept based on the current Zoning strategies available in
facility’s goals (e.g., preservation of the Nigerian giraffe, the Red Book,9 which have been developed based on the
eventual animal movements, and visitation). Your planning proximity to confirmed infected premises.
partners should understand the importance of a goal to the Using USDA zoning classifications (Fig. 9.1), Secure
facility, the species, and the facility mission. Zoo provides suggested Performance Standards, which is
As mentioned, an essential component of planning for an extensive list of response elements for possible inclu-
any FAD event will be the development of a Biosecurity sion in the various biosecurity levels. These Performance
Program. Secure Zoo includes guidance for development Standards are adopted from the other “Secure” programs
of a robust facility Biosecurity Program, which could be and are adapted to address the specific needs of facilities
50 SE C T I O N 1 General
testing methodology, exotic species and indigenous wildlife these documents provide an excellent scientific approach to
pose very different challenges. disposal and decontamination strategies, the extraordinary
Building upon the momentum from existing “Secure” effort involved in carrying out these activities reinforces why
plans, Secure Zoo Strategy supports novel surveillance it is important to prevent a facility from becoming infected
strategies for monitoring animals for disease. Consider the in the first place. The management of an infected facility
recent advances in testing methodologies being used in will not be easy; it may take weeks or months and will be
other species. For example, rather than testing individual extremely expensive. Invest time increasing facility disease
animals in dairies, research has provided a test that may use resiliency; make your facility less vulnerable to infection
a small sample of milk from a dairy farm’s bulk milk tank by continuous improvement in biosecurity in standard
to determine herd negativity, rather than testing individual operating procedures, and be prepared to enhance those
animals in a milking herd. The swine industry is exploring biosecurity measures in the case of an outbreak as discussed
the use of the saliva “rope test” to detect the FMD virus previously (Levels 1 to 3 biosecurity).
on ropes that swine chew on in grouped production set-
tings to avoid having to test individual animals. (J. Tickel,
North Carolina Department of Agriculture and Consumer Contingency Planning Process Step 5:
Services Emergency Programs, personal communication, Plan Preparation, Review, and Approval
January 2017.) Discuss with SAHOs the possible surveil-
lance strategies that would apply to your facility during the This step involves the final writing and organization of
planning process; it is best to understand challenges before a the plan itself. Once drafted, any planning process should
disease outbreak. For facilities that manage native and non- identify who has the authority to review and approve the
native species, the use of domestic sentinel animals should plan, and lay out a schedule for a future plan review. Do not
be considered. A domestic species of low conservation value forget that the staff required to execute the plan should, at
(e.g., a tame steer), which is in contact with valuable suscep- a minimum, be involved in the approval process. Staff may
tible exotic species via fence line exposure or a shared water recognize that elements of a plan may not be achievable at
source, may be easily sampled daily. An agreement with the the operational level either in the short or especially the
SAHO may allow for sentinels to serve as a crucial part of long term, necessitating the need for plan adjustments.
the collections surveillance program and, if negative, confer Not specifically mentioned in the planning process, but
a negative status for the exotic animals in close proximity. critical to success, is the need for training the staff on
Basic facility operations will also need to be discussed their individual roles and responsibilities to execute the
during the planning process. This should include topics plan. Once plans are developed, work with your SAHO to
such as mortality management, disposal, and decontamina- explore training and exercise opportunities that test your
tion. Mortality management is an important discussion to plan in a no-risk, no-fault environment. Consider the devel-
have when planning for an FMD event. Facility owners and opment of a tabletop exercise to test specific elements of
operators should discuss with their SAHO their “priority” your plan; conduct training and drills on the different types
animals for preservation, and determine what might be done of CAPs, and/or perform donning and doffing of personal
to avoid depopulation. Be prepared to explain why these protective equipment, set up footbaths, and try using them
animals are high priority, and be prepared to demonstrate (and keeping them clean) for a day and then a week. With
the facility’s capability of managing these animals should the help of your SAHO, consider participating in state
they become exposed or break with disease. This prioritiza- level exercises, where a scenario includes an exotic animal
tion for preservation pre-event is also helpful information facility in a Control Area, or as an Infected Premises. Most
should vaccination be used as a control option. It is possible states with large agricultural animal populations develop
that vaccination could be employed to halt disease spread these exercises periodically. Find out how your facility might
once the specifics of an FMD outbreak are known, but be included.
initially it would be in limited supply. A facility should
recognize that within its collection, domestic, susceptible Conclusion
livestock or indigenous wildlife of low conservation value
may need to be depopulated if they are not being used as All-hazards contingency planning should be a priority for
sentinels. any facility that manages non-native or indigenous wildlife.
Disposal and decontamination discussions should also Planning efforts protect not only the collection, but also the
occur during the planning process. Plans will need to staff, the public, and the agricultural interests, depending
be flexible and consider as many options as possible, as on the hazard. The success of the planning process will
the tactics that would be employed in a given facility depend on the planning team itself. Carefully consider what
would depend on many variables. Widely used guidelines subject matter experts may assist with risk assessment, plan
for disposal and decontamination may be found in the development, and response. Veterinarians will be important
National Animal Health Emergency Management System’s planning team members; they will be crucial to develop
(NAHEMS) Cleaning and Disinfection10 and Disposal11 plans to prevent disease and should be the point of contact
documents available on the FADPReP website. While for SAHOs, USDA, and other regulatory agencies.
52 SE C T I O N 1 General
Facilities need not “reinvent the wheel” when it comes Guide (CPG) 201: Second Edition, August 2014. https://
to contingency planning. Leverage the expertise of your www.fema.gov/media-library-data/8ca0a9e54dc8b037a55b402b2
planning partners to assist with the steps in the process. a269e94/CPG201_htirag_2nd_edition.pdf. (Accessed January
2017).
No one caring for exotic or indigenous wildlife wants
4. Lloyd ML: Disaster preparation for captive wildlife veterinarians.
to think about the possibility of an FMD outbreak in the In Miller RE, Fowler ME, editors: Zoo and wild animal medicine
United States, but it is critical to consider. The impact (vol 7). St. Louis, 2012, Elsevier, pp 38–46.
on the animals and staff will be incredible, regardless of 5. The ZAHP Fusion Center Web site: Risk Assessment, an annex
the facility’s mission. While this may seem to be a “low from the Zoo Best Practices Working Group for Disaster Prepared-
likelihood” event, every owner and operator should realize ness and Contingency Planning. Available at: http://zahp.aza.org/
that high consequences mean recovery from FMD detection wp-content/uploads/2015/06/Risk_Assessment.pdf. (Accessed
could be a long-term process. January 2017).
In addition to the USDA and applicable State planning 6. The Center for Food Security & Public Health: Secure Food
guidance, Secure Zoo Strategy is available to help facilities Supply Plans. Available at: http://www.cfsph.iastate.edu/Secure-
align themselves (where appropriate) with other “Secure” Food-Supply/. (Accessed January 2017).
7. Secure Zoo Strategy: Available at: http://securezoostrategy.org.
planning efforts. While facility objectives may differ from
(Accessed January 2017).
agricultural animals destined for the dinner table, there 8. United States Department of Agriculture Animal and Plant
are similar goals of continuity and recovery. Secure Zoo Health Inspection Service: FADPReP Materials and Refer-
Strategy recognizes that additional goals of preservation, ences. Available at: https://www.aphis.usda.gov/aphis/ourfocus/
animal transportation, and visitation must be considered animalhealth/emergency-management/ct_fadprep/. (Accessed
in planning efforts for wildlife facilities, and that everyone January 2017).
on the planning team must recognize these goals and work 9. United States Department of Agriculture Animal and Plant
toward individual plans to resume normal operations as Health Inspection Service: FADPReP Foot-and-Mouth Disease
soon as possible. Response Plan: The Red Book. Available at: https://www.aphis
.usda.gov/animal_health/emergency_management/downloads/
fmd_responseplan.pdf. (Accessed January 2017).
References 10. United States Department of Agriculture Animal and Plant Health
Inspection Service: National Animal Health Emergency Manage-
1. Federal Emergency Management Agency, Emergency Manage- ment System Guidelines: Cleaning and Disinfection. Available at:
ment Institute website: IS-230 Fundamentals of Emergency https://www.aphis.usda.gov/animal_health/emergency_manage
Management. Available at: https://emilms.fema.gov/IS230c/. ment/downloads/nahems_guidelines/cleaning_disfection.pdf.
(Accessed January 2017). (Accessed January 2017).
2. Federal Emergency Management Agency, Emergency Manage- 11. United States Department of Agriculture Animal and Plant
ment Institute website: IS-100b Introduction to the Incident Health Inspection Service: National Animal Health Emergency
Command System. Available at: https://emilms.fema.gov/ Management System Guidelines: Disposal. Available at: https://
IS100b/index.htm. (Accessed January 2017). www.aphis.usda.gov/animal_health/emergency_management/
3. Department of Homeland Security Threat and Hazard Identifica- downloads/nahems_guidelines/disposal_nahems.pdf. (Accessed
tion and Risk Assessment Guide, Comprehensive Preparedness January 2017.)
10
Veterinary Occupational Health and
Safety in the Zoo and Wildlife Setting
ELIZABETH E. HAMMOND
53
54 SE C T I O N 1 General
responsibilities under OSHA, which include reading posted together. For instance, chlorine bleach must be kept sepa-
work safety guidelines, complying with OHSA standards, rate from ammonia and acids to prevent the development
abiding by employer safety and health rules/regulations, and of noxious gases.9 Exposure to commonly used chemicals
reporting hazardous conditions and work-related injuries occurs frequently and may lead to adverse reactions. Proper
promptly. PPE and eye wash stations should be readily available. Insec-
A system of reporting injuries should be established so ticides may be a risk to staff, and employers should ensure
that employees know whom to contact (e.g., designated appropriate PPE is available (e.g., fit tested respirators). In
safety officer) in the event of an injury and to ensure proper addition to proper storage and handling, proper disposal
documentation and review of the incident. Any company of hazardous material must be followed. In the United
with 10 or more employees must record any on-the-job States, disposal of these agents may be regulated by the
illnesses and injuries for the calendar year using the OSHA state Environmental Protection Agency, so it is important
300 form as per OSHA regulations.3 A summary of work- to research all federal, state, and local laws.
related injuries and illnesses (OSHA 300A Log) should Because of their potential for abuse, controlled substances
be posted conspicuously at the work site from February are included in the OHSP. In the United States, controlled
1 to April 30 yearly. The safety officer or designee must substances are regulated by the Drug Enforcement Agency
notify OSHA within eight hours of a work-related death (DEA); controlled substances are divided into five categories
and within 24 hours for in-patient hospitalization, ampu- or schedules, but this may vary internationally. Schedule 1
tation, or loss of an eye.4 The OSHA phone number is consists of drugs used for research purposes only, and they
1-800-321-OSHA (6742). have a high level of potential abuse. Schedule 2 to 5 are
Veterinary medicine has inherent safety risks, which may more commonly used by practicing veterinarians. Schedule
be minimized with appropriate measures. According to a 2 drugs, such as morphine, carfentanil, etorphine (M-99),
2011 report, veterinary services ranked 15th in incidence and thiafentanil (A-3080), are more highly regulated than
rates for nonfatal occupational injuries and illnesses.5 As schedule 3 to 5 drugs and have more restrictions on their
such, the American Veterinary Medical Association (AVMA) storage, use, and ordering. A drug register to track the use of
Professional Liability Insurance Trust (PLIT) has published controlled drugs must be kept and available for inspection.
the Veterinary Safety Manual (VSM) to address general In the United States, an inventory of controlled substances
safety in the veterinary workplace.2 Many of the topics should be performed every two years and maintained on
are applicable to zoo and wildlife veterinary medicine, but site for an additional two years.2 Schedule 2 records must
additional issues are specific to this field.6,7 Topics to be be kept separate from schedule 3 to 5. In general, controlled
covered in this chapter include chemical hazards (hazard substances must be locked in a secure cabinet with access
recognition and communication, controlled substances), restricted to a minimal number of employees. If a combi-
physical hazards (sharps, ergonomics, gas cylinders), radia- nation lock is used, the combination should be changed
tion safety, infectious agents (infection control, bloodborne whenever staff leave employment.
pathogens, NHP considerations), and prevention. Detailed An additional consideration for zoo and wildlife vet-
information on additional topics (animal handling and erinarians is the hazard of working with potent narcotics,
restraint, workplace violence, fire and life safety, laser safety) including carfentanil, M-99, and A-3080. Carfentanil is
may be found in the references.2 10,000 times more potent than morphine, and a small
Hazard communication covers the hazardous properties volume can immobilize an elephant.10 Although these drugs
of chemicals in the workplace. OSHA requirements include can be dangerous to humans, antagonists are available.
keeping a list of hazardous chemicals and maintaining Safety Human antagonists (reversals) should always be readily
Data Sheets (SDS, previously known as Material Safety available. When working with potent narcotics, the vet-
Data Sheets [MSDS]). These sheets contain information erinarian should wear appropriate PPE to protect mucous
regarding the hazards of the substance, first aid measures membranes and compromised skin because opioids can be
if exposure occurs, and proper storing and handling of the absorbed at these sites. It is essential to avoid a needle stick
material. The SDSs should be readily available to employ- when working with potent narcotics, so recapping should
ees and periodically reviewed for relevancy. Obsolete files be avoided. A Human Narcotic Safety Protocol should
should be kept for a minimum of 30 years.2 Training of be written in conjunction with local human emergency
employees is an integral part of the hazard communica- personnel that may respond to a potent narcotic expo-
tion plan. For example, it should include information on sure.11 It is important to forge ties with the local emergency
what is considered an exposure, proper work procedure responders and emergency department physicians to alert
(e.g., skin and eye protection when working with chlorine them to the potential for an extreme narcotic exposure.
bleach), and response to accidental exposure (e.g., spill- All staff working around these drugs should be trained on
age of mercury in a thermometer).8 In the United States, the protocol, which should be regularly reviewed. Risks
proper container labeling according to OSHA’s Globally associated with potent opioids and other immobilization
Harmonized System is of utmost importance, especially drugs are addressed in more detail in Chapter 27.
when chemicals are transferred to a different container. It is Radiology with attendant radiation is commonly used
also important to know what chemicals cannot be housed in veterinary medicine, and its regulation may vary among
CHAPTER 10 Veterinary Occupational Health and Safety in the Zoo and Wildlife Setting 55
states or countries. Radiograph equipment vendors may be risks. For any first responders, single-use mouth protection
a good resource for regulations and safe working practices. shields should be provided for mouth-to-mouth resusci-
Routine inspection of equipment and registration of x-ray tation. Waste material soaked in human or NHP blood
generators must be done and may vary according to local or other potentially infectious material (e.g., tissue) is
laws. Radiation detection (dosimetry) badges must be worn regulated waste, and it should be placed in a red biohazard
by staff during all radiation procedures. Dosimetry reports bag and removed by an appropriate disposal company.
are considered medical records and may not be released Contaminated surfaces should be cleaned and sanitized.
without written consent. Dosimetry badges are evaluated If exposure to infectious material is suspected, the affected
quarterly for the level of exposure to radiation using the area should be washed with soap and water immediately,
“ALARA” notification levels (ALARA = As Low As Reason- and the supervisor and safety officer should be notified. The
ably Achievable) in the United States.2 As per OSHA, the employee may be directed to a physician for evaluation or
following regulations should be followed: individuals under treatment.
18 years of age are not permitted to work with radiograph Prevention of hepatitis B is another important aspect of
equipment, leaded aprons with thyroid protection greater the BBP OHSP. In the United States, employees recognized
than or equal to 0.25 mm lead and hand protection greater at potential risk for exposure to human bodily fluids while
than or equal to 0.5 mm lead must be made available.2 The performing their work duties should be offered the hepatitis
operator must stand at least 6 feet away unless the animal B vaccine within 10 days of employment at no cost to them
must be held by the operator, and users should avoid the as per OSHA.2 A declination form should be signed by
direct beam of radiation. A record of all staff training should the employee if he or she opts out of the vaccination and
be kept, and annual refresher safety training should be should be kept in the employee’s file.15 In addition, OSHA
performed. requires the employer to keep a confidential record of the
Infection control is another component of the OHSP, employee’s date of hepatitis B vaccination and a copy of any
and handwashing is the most important way to prevent the potential exposure to any infectious material, treatment,
spread of disease. Hands should be washed with soap for a and medical procedures. These records must be kept for the
minimum of 15 seconds. Warm water may increase compli- entire length of an employee’s employment plus 30 years.2
ance, but cold water is equally effective. Soap should be dis- Additional information on hepatitis B vaccination is found
pensed from a disposable container because refillable soap in Table 10.1.
dispensers may harbor infectious organisms.12 Hands-free Staff training on zoonotic diseases should also be
faucets, soap, and towel dispensers may be advantageous. documented in the employee’s record. For employees who
When running water is not available, 60%–95% ethyl alco- potentially have access to animals in a zoo or wildlife
hol or isopropyl alcohol-based hand sanitizers are very effec- setting, it is recommended that they have a pre-employment
tive once the gross debris has been removed. However, they physical exam, intradermal skin test for tuberculosis, fecal
do not eliminate bacterial spores (e.g., clostridial spores) or examination, and blood drawn so that a 5 mL serum
cryptosporidium and are not as effective against nonenvel- sample (divided into two aliquots) may be stored at −20°C.6
oped viruses (e.g., parvovirus).13 Fingernails should be kept However, privacy laws and expenses associated with storing
short, and jewelry should not be worn in order to maximize medical samples may discourage facilities from collect-
hand hygiene. Eating, drinking, and smoking in labora- ing this information. It is best to consult with the HR
tory or animal areas should not be allowed, and employ- department, the institution’s attorney, the employee’s union
ees should be offered a separate area for these activities. (where applicable), and a physician prior to establishing a
Dedicated work clothes and shoes are an important part of serum bank on site.16 A list of recommended vaccinations
disease prevention in the veterinary setting. Work clothes for employees may be found in Table 10.1. In addition to
should be laundered at the work site using hot water and rabies and tetanus prophylaxis, employees working with
drying at a high temperature. This is especially important NHPs should have proof of measles, hepatitis A and B, and
when working with infectious agents and NHPs. poliomyelitis prophylaxis.16
Bloodborne pathogen (BBP) safety of the OHSP was Although many zoonotic diseases are relevant to the zoo
designed to protect employees from human immunodefi- and wildlife veterinarian, this chapter is limited to several
ciency virus (HIV) and hepatitis B, and it relates to anyone important diseases. Rabies is a potentially fatal disease
who may reasonably come into contact with human bodily caused by viruses in the genus Lyssavirus. Any mammal may
fluids during the course of his or her work duties. Janito- potentially transmit the virus, although certain animals, such
rial workers and first aid responders fall into this category. as bats, are known reservoirs for the disease. Pre-exposure
Because NHPs may also carry zoonotic infectious diseases, rabies prophylaxis is recommended for any personnel
zoo and wildlife veterinarians and support staff who work working with a potentially rabid animal. In the United
with these species may be exposed to BBP.14 Identifying States, most states require that mammal bites be reported
potential exposure to BBP and instituting control measures to the health department and animal control, although
are the first steps in developing this aspect of the OHSP. this may vary by state and local laws. Prompt treatment of
Providing appropriate PPE (e.g., safety glasses, disposable, a bite wound and post-exposure treatment greatly increase
liquid-proof gloves) and training are critical to minimize the chances of survival of a victim of a rabid animal bite.
56 SE C T I O N 1 General
TABLE
10.1 Recommended Vaccination and Testing for Employees in a Zoo and Wildlife Setting*
Psittacosis is the disease caused by the Chlamydia psittaci should be suspected in any nonhealing wound. If an animal
bacterial organism. It may be treated with antibiotics, and with a wound is determined to have MRSA based on
human cases are rarely fatal.17 OSHA recommendations to culture and sensitivity, it is best to alert all involved in the
protect workers from contracting psittacosis through inha- care of that animal.19 Appropriate PPE should be instituted
lation include providing appropriate respiratory protection based on the facility and potential exposure. As a general
(e.g., high efficiency particulate air [HEPA] filter), establish- rule, dedicated boots, gowns, and disposable gloves should
ing a respirator program and a psittacosis training and pre- be worn around the MRSA-positive animal. In addition,
vention program, and ensuring adequate ventilation in the any caretakers with nonhealing wounds should alert their
work area.18 This program should educate staff on the clini- physician to the fact that they have been caring for an
cal signs associated with psittacosis in avian species. Keepers animal with MRSA. For shared computers, keyboard covers
should immediately report to their supervisor any indica- that are easily disinfected may reduce the risk of spreading
tions that a bird may be infected, which may include ocu- MRSA or other infectious organisms in the workplace.
lonasal discharge, conjunctivitis, and weight loss. However, Toxoplasmosis is a disease caused by the protozoan
many birds may be inapparent carriers. In addition, if an Toxoplasma gondii. Felids are the definitive hosts for this
employee is showing flu-like symptoms (fever, malaise, head- pathogen, which is found worldwide. In the immuno-
ache), the employee should alert the physician to the fact competent person, infection may be subclinical, but
that he or she works with birds.17 Confirmed work-related immunocompromised people may be at risk of developing
cases of psittacosis must be reported to OSHA, the state severe disease. Pregnant women should avoid exposure to T.
health department, and the National Centers for Disease gondii because it can cause serious birth defects in the fetus.
Control and Prevention in the United States.18 The list of Serologic titers should be performed on women of child-
reportable diseases should be reviewed in other countries. bearing age to determine serologic status. Pregnant women
Methicillin-resistant Staphylococcus aureus (MRSA) is a working around felids should avoid exposure to feline feces
potentially highly resistant version of a relatively ubiquitous or soil contaminated with feline feces.16 Job reassignment
organism, and many humans are carriers. This organism should be considered in those workers considered high risk.
CHAPTER 10 Veterinary Occupational Health and Safety in the Zoo and Wildlife Setting 57
Reptiles and amphibians are often carriers for Salmonella With this method, one hand is used to scoop up the cap
sp. Therefore, anyone working with these animals should be over the needle, and the cap is depressed over the needle by
informed of the potential risks of this disease and instructed pressing it against a hard object, such as the wall. Needles
on precautions that should be taken to prevent it. Birds with a recapping device are commonly used in human
and mammals may also carry salmonella, so any animal in medicine and may be a good, safe option in veterinary
contact with the public should also be screened for salmo- medicine. Needle caps should never be placed in the mouth.
nella on a regular basis.20 Hand-washing after handling any Gross necropsies are an integral part of preventative
of these animals is the best way to prevent disease. medicine and zoo and wildlife health programs, but they
Because humans and NHPs share many diseases, present a potential risk of pathogen exposure and physical
guidelines for disease prevention from the human health injury. At a minimum, latex gloves, protective outwear (e.g.,
care sector may be applicable when working with NHPs. coveralls or apron), eye protection, and face mask should be
Identification of risks may depend on the history, health, worn. Veterinarians performing necropsies may be exposed
and species of the NHP in the collection, facility design, to formaldehyde, which is an inhalant hazard and carcino-
among other factors. There are varying levels of exposure gen.23 Knife injuries during necropsies are common; 87%
to NHPs. For instance, cleaning a contaminated cage may of respondents in a survey of zoo veterinarians reported a
expose the worker to aerosolization and direct contact of knife wound during a necropsy and half of those wounds
potential pathogens, whereas maintaining a greater than required medical treatment.7 Measures to prevent knife
one meter distance from an NHP carries a lower risk. When wounds, such as the use of washable, cut-proof gloves,
working with NHPs, minimum PPE should include a face should be considered where practical. When power tools
mask, gloves, long sleeves, and pants, although additional are used, it is important to wear appropriate eye protection
precautions should be taken when working with macaque and a respirator. Training on power tool or other heavy
species.21 Coveralls or similar outer protective wear and equipment use should be mandatory and documented. In
dedicated boots are also recommended. For more direct addition, injuries often happen when workers are fatigued,
contact with NHPs and their bodily fluids, such as per- so it is important to remind workers to slow down.
forming dentistry on an anesthetized animal or cleaning The OHSP also covers ergonomics, which prevents
enclosures, face shields and hair covering should also be work-related musculoskeletal disorders in employees.
worn. In hot climates, the use of PPE may cause the wearer Employers should provide proper computer working
to overheat; providing spots to cool down and maintaining stations to prevent back or wrist strain, eye fatigue, and
hydration may prevent heat sickness in employees. Anyone repetitive motion injuries.2 In addition, employees should
showing signs of illness, such as upper respiratory infection, be reminded of safe lifting practices to reduce back injury.
cold sores, or gastrointestinal upset, should avoid working A 1998 survey of zoo veterinarians revealed that over half
with NHPs until clinical signs have resolved. Clothing worn of respondents reported a work-related back injury.7
in NHP areas should not be worn outside of that area and Many veterinary clinics use compressed gas cylinders,
ideally should be laundered on site. Use of foot baths with which can be very dangerous if mishandled. The manufac-
appropriate disinfectants is encouraged to minimize transfer turer’s instructions should be followed. The cylinders should
of pathogens from NHP enclosures. be stored in an upright position and secured in place, and
Herpes B virus (Herpesvirus simiae, Macacine herpesvirus, a protective cap should cover the valve when not in use.
Cercopithecine herpesvirus 1) is often fatal in humans. It is The cylinders should not be stored in a warm environment,
common in macaques, who are inapparent carriers. Limit- exposed to corrosive substances, or transported with the
ing exposure, safe handling procedures, and more extensive regulator attached.2
PPE, such as eye protection, should be worn when working Additional occupational hazards for zoo and wildlife vet-
with macaques or animals known to be infected with herpes erinarians include venomous reptiles and dangerous animals
B. Prompt treatment of bites or scratches is essential. A bite/ (e.g., elephants), but a detailed discussion is beyond the
injury protocol should be available in all primate areas.22 scope of this chapter.11,24 With proper training and docu-
It is advisable to discuss in advance primate bite protocols mentation, occupational hazards for the zoo and wildlife
with the hospital staff that may be asked to address them. veterinarian may be minimized, and the veterinarian can
Educating staff about the risks of the disease and how to be a great resource for the facility’s OHSP.
prevent it are paramount. Other viruses of NHPs, such as
STLV, SIV, GaLV, may present potential zoonotic risks.14,22 Acknowledgments
In addition to exposure to BBP, injuries associated with
“sharps” (needles, scalpel blades, broken glass vials, etc.) A heartfelt thank you goes to Drs. Holly Haefele, Genevieve
should be included in the OHSP. Safety measures should be Dumonceaux, and Michele Miller for reviewing this chapter.
taken when using sharp objects, and they should be placed
in an appropriate sharps container immediately after use. References
Needles should not be recapped but immediately disposed
of in an appropriate sharps container. However, if recapping 1. United States Department of Labor: Occupational Safety and
is necessary, the one-hand recap method should be used. Health Administration, 2017. Retrieved from www.osha.gov.
58 SE C T I O N 1 General
2. AVMA PLIT Veterinary Safety Manual: HUB international 15. United States Department of Labor: OSHA: Health Care Pro-
midwest limited, Chicago, IL, 2009, p 204. fessionals Hepatitis B Declination Statement, 2017. Retrieved
3. United States Department of Labor: OSHA: Injury & Illness from https://www.osha.gov/SLTC/etools/hospital/hazards/bbp/
Recordkeeping Forms – 300, 300A, 301, 2017. Retrieved from declination.html. (Accessed 14 February 2017).
https://www.osha.gov/recordkeeping/RKforms.html. (Accessed 16. Shellabarger WC: Appendix 6: Zoo personnel health program
14 February 2017). recommendations. In the Guidelines for Zoo and Aquarium
4. United States Department of Labor: OSHA: Report a Fatality Veterinary Medical Programs and Veterinary Hospitals. Veteri-
or Severe Injury, 2017. Retrieved from https://www.osha.gov/ nary Standards Committee, 1998. American Association of Zoo
report.html. (Accessed 26 January 2017). Veterinarians. (Accessed 15 February 2017).
5. Centers for Disease Control and Prevention: The National Insti- 17. Smith KA, Campbell CT, Murphy J, et al: Compendium of
tute for Occupational Safety and Health (NIOSH): Veterinary measures to control Chlamydophila psittaci infection among
Safety and Health, 2012. Retrieved from https://www.cdc.gov/ humans (psittacosis) and pet birds (avian chlamydiosis), 2010.
niosh/topics/veterinary/work.html. (Accessed 26 January 2017). http://www.nasphv.org/Documents/Psittacosis.pdf. (Accessed 14
6. Silberman MS: Animal and human welfare: occupation health February 2017).
programs in wildlife facilities. In Fowler ME, editor: Zoo and 18. OHSA hazard information bulletins contracting occupationally
wild animal medicine: current therapy, ed 3, Philadelphia, 1993, related psittacosis, 1994. Retrieved from https://www.osha.gov/
W.B. Saunders Company, pp 57–61. dts/hib/hib_data/hib19940808.html. (Accessed 10 March 2017).
7. Hill DJ, Langley RL, Morrow WM: Occupational injuries and 19. Methicillin-Resistant Staphylococcus aureus skin infections from
illnesses reported by zoo veterinarians in the United States, in an elephant calf — San Diego, California, 2008. Retrieved from
Proceedings, AAZV and AAWV Joint Conference 484–488, 1998. https://www.cdc.gov/mmwr/preview/mmwrhtml/mm5808a3
8. United States Department of Labor: OSHA: Overview, .htm. (Accessed 16 February 2017).
2017. Retrieved from https://www.osha.gov/SLTC/mercury/ 20. Compendium of measures to prevent disease associated with
index.html. (Accessed 14 February 2017). animals in public settings, 2013; NASPHV animal contact
9. Centers for Disease Control and Prevention: Emergency compendium committee, J Am Vet Med Assoc 243:1270–1288,
Preparedness and Response: Chlorine (Cl), 2013. Retrieved 2013.
from https://emergency.cdc.gov/agent/chlorine/basics/facts.asp. 21. Occupational primate disease safety guidelines for zoological
(Accessed 14 February 2017). institutions. Retrieved from http://www.aazv.org/?page=12.
10. Caulkett N, Shury T: Human safety during wildlife capture. (Accessed 17 February 2017).
In West G, Heard D, Caulkett N, editors: Zoo animal and 22. Murphy HW, Switzer WM: Occupational exposure to zoonotic
wildlife immobilization and anesthesia, ed 2, Ames, 2014, Wiley simian retroviruses: health and safety implications for persons
Blackwell, pp 181–187. working with nonhuman primates. In Fowler ME, Miller RE,
11. Zoo and aquarium safety: example practices, 2015. Available editors: Zoo and wild animal medicine: current therapy, ed 6, St.
at: https://www.aza.org/safety-security. (Accessed 14 February Louis, 2008, Saunders Elsevier, pp 251–264.
2017). 23. Occupational Safety and Health Administration: Laboratory
12. NASPHV Compendium of veterinary standard precautions, Safety Guidance, 2011. Retrieved from https://www.osha.gov/
J Am Vet Med Assoc 247:1252–1277, 2015. Publications/laboratory/OSHA3404laboratory-safety-guidance
13. Infection prevention and control best practices for small animal .pdf. (Accessed 21 February 2017).
veterinary clinics, August 2008. www.wormsandgermsblog.com/ 24. Flanagan JP: Snakebite protocols for zoos. In Fowler ME, Miller
files/2008/04/CCAR-Guidelines-Final2.pdf. (Accessed 17 Feb- RE, editors: Zoo and wild animal medicine: current therapy,
ruary 2017). ed 4, Philadelphia, 1999, W.B. Saunders Company, pp 95–100.
14. Roberts JA: Occupational health concerns with nonhuman
primates in zoological gardens, J Zoo Wildl Med 26(1):10–23,
1995.
11
Research Study Design
NICOLA DI GIROLAMO AND CHRISTOPH MANS
59
60 SE C T I O N 1 General
• Figure 11.1 Simplified algorithm for the determination of the study design of a research study.
anesthesia, etc.) not directly related to the tested interven- 2004 and 2008. The study concluded that the addition
tion. Caregivers should be blinded to the intervention (e.g., of the fenbendazole improved the outcomes of rabbits.9
drug) administered at each time point to avoid a possible However, this study design may be affected by multiple
differential care of the animals. Outcome assessors should types of bias because a general improvement of the care
be blinded to the intervention administered at each time of the animals could be also responsible for the improved
point to avoid biased assessments. outcomes. In nonrandomized studies, confounding
factors need to be addressed by using proper statistical
Nonrandomized Studies techniques that account for multiple predictors, such as
general linear model, generalized linear model, and logistic
Nonrandomized studies provide limited evidence of the regression.
effects of interventions. They are the main source of evidence
on the intended effects of many organizational or public Study Designs for Identification of
health interventions and on interventions that cannot
ethically be randomized. A large class of nonrandomized Underlying Disease Etiologies
studies includes those in which allocation occurs based on Cohort Studies
predefined groups or on clinical decisions, such as those of
the treating clinician, the curators of the animals, or the A cohort study is a type of observational study in which a
animal owners. These observational studies are many of group of animals that share an exposure (e.g., a defining
the classical epidemiologic designs such as cohort studies, characteristic, or a common event in a selected period)
case-control studies, and cross-sectional studies. However, is compared with a group of animals similar in all the
these study designs are better fitted for identification of other characteristics but that did not have such exposure.
underlying disease etiologies. Other nonrandomized studies Briefly, if the exposed group develops a higher incidence
compare a group of animals receiving an intervention of the outcome than the unexposed, then the exposure
with a group of animals of the past that did not receive is associated with an increased risk of the outcome.10 The
such intervention. A clear example of such “historically strength of cohort studies is that they enable calculation of
controlled studies” is a study in which rabbits affected by incidence rates, relative risks, and attributable risks. The
Encephalitozoon cuniculi were treated without fenbendazole main weakness of cohort study is that for outcomes that
between 2000 and 2003 and with fenbendazole between are rare or develop over longer time periods, this type of
CHAPTER 11 Research Study Design 61
research design can be slow and expensive. Selection and test as true positive, false positive, true negative, and false
information bias are a concern with cohort studies. negative.13 This information is critical for clinicians and
allows for an unbiased evaluation of the diagnostic test
Case-Control Studies results.
9. Sieg J, Hein J, Jass A, et al: Clinical evaluation of therapeutic 13. Mallett S, Halligan S, Thompson M, et al: Interpreting
success in rabbits with suspected encephalitozoonosis, Vet Para- diagnostic accuracy studies for patient care, BMJ 345:e3999,
sitol 187(1–2):328–332, 2012. 2012.
10. Grimes DA, Schulz KF: An overview of clinical research: the lay 14. Bland JM, Altman DG: Statistical methods for assessing agree-
of the land, Lancet 359(9300):57–61, 2002. ment between two methods of clinical measurement, Lancet
11. Higgins JP, Ramsay C, Reeves BC, et al: Issues relating to study 1(8476):307–310, 1986.
design and risk of bias when including non-randomized studies 15. Friedrichs KR, Harr KE, Freeman KP, et al: ASVCP reference
in systematic reviews on the effects of interventions, Res Synth interval guidelines: determination of de novo reference intervals
Methods 4(1):12–25, 2013. in veterinary species and other related topics, Vet Clin Pathol
12. Borowsky A, Esserman L: When the gold standard loses its luster, 41(4):441–453, 2012.
perhaps it is time to change nomenclature, Ann Intern Med
164(10):694–695, 2016, doi:10.7326/M16-0526. Epub 2016
Mar 22.
SECTION 2
Animal Welfare
12 Overview of Animal Welfare in Zoos, 64
63
12
Overview of Animal Welfare in Zoos
JOANNE PAUL-MURPHY AND CHR ISTINE MOLTER
Introduction to Animal Welfare in Zoos the animal, and that it impacts survival and must be viewed
from their perspective. Welfare refers to a characteristic of
The mission of zoos and aquariums is broad and emphasizes the individual animal rather than something given to the
achievements in areas of animal care, conservation, education, animal by people.7 Welfare is about the subjective state of
science, and recreation. A critical element of that mission the animal, including the judgment of what is good in the
is to constantly strive toward achieving higher standards of animal’s life and what is not good. Because welfare belongs
welfare for wild animals, for captive animals directly under to the individual animal, it will differ between different
their stewardship, and for those still free-ranging or under animals, even when the exact same conditions are provided,
in situ management. Nonhuman animal welfare (from here and the welfare of the same animal will change as the animal
forward termed “animal welfare”) is vital to supporting the ages. In the case of zoos, animals may have come from a
goals of zoos and aquariums (from here forward termed variety of backgrounds, individuals may vary greatly in their
“zoos”) as modern conservation organizations.1 previous life experiences, and this may influence their ability
Zoos are united through organizations, including the to cope with certain challenges. For example, an animal that
World Association of Zoos and Aquariums (WAZA), the stays in zoos its whole life requires different skills from an
Association of Zoos and Aquariums (AZA), the European animal that is kept for the future goal of reintroduction into
Association of Zoos and Aquaria (EAZA), and other regional the natural environment or another example of an animal
zoo organizations, that provide support and channels of brought into an urban zoo from a semi-free-ranging facility
communication to maintain welfare and ethical standards; or vice versa.8,9 By using each animal as its own control, an
however, a universally accepted definition for animal welfare individual’s welfare can be tracked in response to changes
has not been agreed upon in the zoo or in other professional in its environment and, thus, an individual’s welfare can be
communities, and several entities have developed their own measured. The welfare assessment of an animal or a group
definitions and descriptions (Table 12.1).2 of animals may result in different conclusions, depending
Acceptable welfare standards and best practices remain on the goal of keeping an animal.
somewhat vague and elusive; they are the subject of much Moreover, zoos are acutely aware of the public’s percep-
debate.3 This may be a result of differing ideas about what tion of the well-being of captive animals. Many zoos have
animal welfare actually is and the continuum of zoological a high degree of public approval and acceptance, especially
institutions, from zoos and wildlife parks to game reserves, when compared to other institutions caring for captive
national parks, and protected areas, thus making the idea animals for research or production. At the same time, zoos
of enclosure and captivity fluid and not absolute.4 This are under constant criticism from the public because people
is coupled with the relatively young and extremely broad have complete access to directly observe how the captive
discipline of zoo animal welfare science grappling with a animals are behaving. It is the responsibility of the zoo
variety of approaches to the study of welfare, the difficulty community to retain the public’s regard by understanding
of conducting scientific studies in zoos, and a lack of the changing societal consensus about animals, and by
information about many animals, both in captivity and designing zoo environments that meet the needs that the
in the wild.5 Like any science, welfare studies are being animal itself perceives to be important.10
continuously advanced, informed, and debated by newer
investigations, and are refined as they progress. Emerging
research in the area of what constitutes good animal welfare Animal Welfare Assessments in Zoos
rather than identification of what needs to be avoided may Establishing Practical Guidelines
yield constructive information to design best practices.6 for Assessments
Despite various definitions, multidimensional aspects,
and interdisciplinary principles of animal welfare, it is Assessing and maintaining welfare for an individual animal
important to keep in mind that animal welfare belongs to or a population of animals is the responsibility of everyone
64
CHAPTER 12 Overview of Animal Welfare in Zoos 65
TABLE the person who has daily interactions with the animal
12.1 Various Definitions of Animal Welfare and will notice subtle changes in vitality, activity,
behavior, and food intake.
American College Animal welfare refers to the state
of Animal of the animal. Assessment of b. Veterinarians—these are the people with special
Welfare44 welfare includes consideration of training to observe and examine the animal’s health
the animal’s health, behavior, and parameters and should also be familiar with physical
biological function attributes specific to the species under evaluation.
American Welfare means overall mental and c. Animal curators—these are the people who are
Veterinary physical health; protecting an trained to observe a variety of animal species in a
Medical animal’s welfare means providing zoo, and they are knowledgeable about requirements
Association33 for its mental and physical needs
and compliance with guidelines or protocols set by
Association of Animal welfare refers to an animal’s government agencies, professional organizations, and
Zoos and collective physical, mental, and population management plans.
Aquariums25 emotional states over a period
d. Research investigators—these are the people who may
of time, and is measured on a
continuum from good to poor have zoo animals involved in any type of study. The
investigators may be collecting specific information
World Animal welfare means how an animal
that changes over time relative to the animal’s welfare.
Organization is coping with the conditions
for Animal in which it lives. An animal is When captive animals are engaged in studies, it is often
Health (OIE)49 in a good state of welfare if (as necessary to have an institutional animal care and use
indicated by scientific evidence) protocol reviewed by the Institutional Animal Care
it is healthy, comfortable, well- and Use Committee (IACUC) or internal research
nourished, safe, able to express
review panel. This review may initiate a welfare assess-
innate behavior, and if it is not
suffering from unpleasant states, ment protocol early in the planning of a project.
such as pain, fear, and distress 2. Good communication and records of the observations
will aid in reducing the range of variability. Consistency
between observers, or at least between the veterinary
working within a zoo. Everyone has a role to ensure that professionals participating in a welfare assessment of the
animals are thriving. An institutional welfare assessment same animals or facility, improves the follow-through
program and welfare committee provide support for the and re-evaluation. There is predictable variation between
individuals caring for the animals. Additionally, some observers; therefore, having the same people observing
animal welfare departments have responsibilities for animal the animals for reevaluation is helpful. Furthermore,
training, behavioral husbandry, enrichment, and research. individual animals may be able to differentiate between
A proper welfare assessment must draw on multiple the observers and display different behaviors depend-
disciplines and consider that the perspectives of the human ing on the level of familiarity with the person making
scientists, caretakers, or policymakers often result in dif- observations. When animals are enrolled in studies or
ferent emphases on what constitutes a welfare problem or research projects, it is important that all team members
where efforts should be focused to improve the situation.11 understand the purpose of the study and the scientific
Each individual evaluating or contributing to a welfare objectives.
assessment brings his or her own biases and ethical views, 3. Identification of which indicators are to be monitored
which affects how different elements of welfare are weighted can be challenging, including which behavioral and
in an assessment. When developing a welfare assessment physiologic parameters are used as indicators of welfare
for animals in a zoological institution, there are several (see Chapter 13). It is important to define and monitor
over-reaching principles that have been established that can the right types and number of indicators. Common errors
be applied. The following set of recommendations has been include having too many parameters, which prolongs the
extrapolated from animal welfare assessment guidelines for process and makes it less effective, whereas too few may
laboratory animals.12 lead to inaccuracies. Setting the baseline is often the
1. A team approach allows input from people with different most challenging part of a welfare assessment because
perspectives of the animal’s situation. Identification of it involves agreement as to what may be considered the
who is involved with an animal care team will vary. An hypothetical ideal.
assessment often receives input from several different 4. The record keeping system for welfare information col-
individuals, and it works best when people in the team lected should be based on what is appropriate for the
are prepared to work together. It is beneficial when all institution being evaluated. There are several systems
members of the team are familiar with normal behavior available for recording welfare assessments in zoos,
for the species being evaluated and are able to recognize including developing modules within existing electronic
abnormal behaviors. medical records systems or independent applications,
a. Animal caregivers—these are the people who observe such as WelfareTrak (Chicago Zoological Society,
the animal in the captive environment. This is often Brookfield, Illinois), as well as systems utilized in food
66 SE C T I O N 2 Animal Welfare
or laboratory animal facilities.13 Record systems often the animals in a malnourished condition or in a state of
have predetermined lists of factors to be evaluated. These over-nutrition. An assessment gives consideration to the
may be set up to answer with simple binary (yes/no) presentation of the food, the variability in food types, and
responses or may incorporate numerical or ranking scales the feeding schedule, or giving the animals choice and
from good to bad. Numerical scoring systems provide control of food items. Resource-based measures are used in
structure to evaluate clinical signs, physical indicators, conjunction with information about management practices
and behavioral parameters; however, this system requires to help identify the causes of animal welfare problems or
a subjective value judgment by each assessor. indicate potential risk factors.16 Exhibit design must incor-
5. Frequency and timing of welfare assessments are often porate species-specific needs and appropriate space alloca-
determined by the degree of deviation from good tions, plus provide environmental enrichment features and
welfare. Assessments will vary depending on each situa- opportunities for natural locomotion, exercise, and suitable
tion, and the timing and frequency is often established social interactions with other animals in the exhibit. It is
by the number of animals involved and the allocation of often the housing and management conditions for animals
resources to provide effective monitoring. Indicators of maintained by an institution that are scrutinized by the
poor welfare should be determined first and addressed public. Resources are necessary components of welfare,
immediately. After specific thresholds are determined, ensuring that conditions are present that provide animals
indicators of good welfare can then be used for monitor- with the potential to experience good welfare, but they do
ing. Signs of poor welfare may be more challenging not—in and of themselves—ensure good welfare.5
to detect in some species, which could lead to more Animal-based (also termed “outcome-based” or “out
frequent assessments in hopes of identifying problems puts”) measures include the physical, behavioral, and mental
early. The age or condition of illness of an animal will state of the animals themselves. Because welfare needs to
also affect the frequency of health and welfare assess- be evaluated from the animals’ perspectives, animal-based
ments. With regard to animals involved in a study, the measures are critical; however, some of these, such as the
experimental design usually affects the frequency and mental state, can be scientifically challenging to measure.
timing of observations. Signs of illness and pain are Evaluation of the animal’s physical state is usually the area
important initial considerations that would prompt an of greatest familiarity for veterinarians because it includes
immediate corrective action if such signs were unex- the health of the animal. The physical state assessment
pected, or unrelated to the research.12,14 includes information provided by the caregiver, such as food
and water consumption, feces and urine or urate produc-
Animal Welfare Assessment Principles tion, and level of activity. Evaluation includes the objective
measurements of a physical examination such as weight,
Methods to perform an animal welfare assessment may heart rate, respiratory rate, and recovery time after handling.
be found in textbooks and peer-reviewed publications, Additional information to assess the physical state of an
although the primary subjects are often laboratory or animal frequently includes diagnostic evaluations, such as
production animals. Specific recommendations for how the complete blood count (CBC), plasma biochemistry
to perform welfare assessment of animals living in zoos values, fecal evaluation, and urinalysis. Radiographic and
have only recently emerged in the literature, and several additional imaging studies may be included. Physical assess-
guiding principles or models have been set forth (Table ments also include several subjectively scored parameters,
12.2). An assessment includes an evaluation of both positive such as the animal’s degree of responsiveness, body condi-
and negative welfare states; it is best when measured on a tion score, mobility, and posture.
sliding scale from very poor to very good, and should be Many animal welfare studies examine subclinical physi-
measured scientifically whenever possible. One approach ologic changes to determine if an animal is distressed, when
to organizing a welfare assessment of animals in zoos is the stress response shifts sufficient resources to impair
to include measurements of two major components, both other biological functions.17 An example would be a bird
animal based and resource based.15 experiencing stress in a new enclosure that doesn’t show
Resource-based (also termed “design-based” or “inputs”) outward signs but may become immunocompromised and
reviews are the most commonly applied evaluation of an susceptible to fungal infection. Glucocorticoid measurement
animal’s welfare assessment. Resource-based measures focus using blood, feces, or saliva has been used as an indication of
on the animal care and things that are provided to the adrenal function and the animal’s stress response; however,
animals, with natural history taken into account, such as interpretation of glucococorticoid metabolites concentra-
the amount of space, substrate, temperature, diet, and vet- tions requires caution because it is affected by a long list
erinary care. Some welfare resources are measured by being of variables, not necessarily stressful or distressing, such as
either present or not present, such as providing the correct diurnal or seasonal rhythms, habituation, diet, sex, and
thermal zone (positive) or being outside of that thermal reproduction (see Chapter 13).18
zone (negative), whereas other parameters are evaluated on Assessment of the behavioral state evaluates whether an
a continuum; for example, sufficient food may be provided animal is engaging in activities typical for the species and
and the animals are not hungry; however, the diet may not expressing maladaptive behaviors that result in injury
not be providing the correct nutritional balance, placing or illness. The behavior of captive animals is frequently
CHAPTER 12 Overview of Animal Welfare in Zoos 67
TABLE
12.2 Guiding Principles for Assessing Animal Welfare
American Veterinary Medical 1. The responsible use of animals for human purposes, such as companionship, food, fiber,
Association: recreation, work, education, exhibition, and research conducted for the benefit of both
Eight integrated principles for humans and animals, is consistent with the Veterinarian’s Oath.
developing and evaluating 2. Decisions regarding animal care, use, and welfare shall be made by balancing scientific
animal welfare policies, knowledge and professional judgment with consideration of ethical and societal values.
resolutions, and actions33 3. Animals must be provided water, food, proper handling, health care, and an environment
appropriate to their care and use, with thoughtful consideration for their species-typical
biology and behavior.
4. Animals should be cared for in ways that minimize fear, pain, stress, and suffering.
5. Procedures related to animal housing, management, care, and use should be continuously
evaluated, and when indicated, refined or replaced.
6. Conservation and management of animal populations should be humane, socially
responsible, and scientifically prudent.
7. Animals shall be treated with respect and dignity throughout their lives and, when
necessary, provided a humane death.
8. The veterinary profession shall continually strive to improve animal health and welfare
through scientific research, education, collaboration, advocacy, and the development of
legislation and regulations.
Five Freedoms Model50 1. Freedom from hunger and thirst by ready access to fresh water and a diet to maintain full
health and vigor
2. Freedom from discomfort by providing appropriate environment including shelter and a
comfortable resting area
3. Freedom from pain, injury, or disease by prevention or rapid diagnosis and treatment
4. Freedom to express normal behavior by providing sufficient space, proper facilities, and
company of the animal’s own kind
5. Freedom from fear and distress by ensuring conditions and treatment that avoid mental
suffering
Animal Welfare51 1. Need for a suitable environment
2. Need for a suitable diet
3. Need to be able to exhibit normal behavior patterns
4. Need for the company of, or to be apart from, other animals
5. Need to be protected against pain, suffering, injury, and disease
Five Domains Model1,52 Animal welfare is divided into physical and functional domains (nutrition, environmental, physical
health, behavior) and mental domain (negative and positive experiences). Both internal
and external conditions give rise to negative (aversive) and positive (pleasant) subjective
experiences, the integrated effects of which give rise to the animal’s welfare status.
Maslow’s Hierarchy of Needs Animal welfare should be directed toward the highest categories of Maslow’s pyramid of
Pyramid1 wellness and well-being. The bottom of the pyramid includes the critical foundational
requirements for survival, including physiologic needs for shelter, water, and hygiene, as
understood through experience and science. The middle of the pyramid includes the
animals’ physical health and safety needs. The top of the pyramid is the most varied and
complex welfare-related activities, such as social needs, mental stimulation, and choice. This
pyramid, when all parts are achieved, results in an animal retaining and encouraging natural
abilities.
Opportunities to Thrive27,57 1. Opportunity for a well-balanced diet: fresh water and a suitable, species-specific diet will be
provided in a way that ensures full health and vigor, both behaviorally and physically
2. Opportunity to self-maintain: an appropriate environment including shelter and species-
specific substrates that encourage opportunities to self-maintain
3. Opportunity for optimal health: providing supportive environments that increase the
likelihood of healthy individuals as well as rapid diagnosis and treatment of injury or disease
4. Opportunity to express species-specific behavior: quality spaces and appropriate social
groupings will be provided that encourage species-specific behaviors at natural frequencies
and of appropriate diversity while meeting social and developmental needs of each species
in the collection
5. Opportunities for choice and control: providing conditions in which animals can exercise
control and make choices to avoid suffering and distress, and make behavior meaningful
68 SE C T I O N 2 Animal Welfare
compared to the behavior of their wild counterparts to the primary purposes of a zoo welfare committee is the
understand the effects of captivity.5 The possibility that oversight of welfare assessments. The committee may not
animals in captivity can perform behaviors that they be the team tasked with performing the assessment, but
would normally perform in a more natural environment may request welfare assessments in response to concerns
is an enduring welfare concern, and seemingly a higher presented, to identify strengths and areas in need of
priority for zoo animals (never considered tame) than advancements, to create a system or process to evaluate the
for domesticated animals. Positive welfare exists when an welfare, to start scientific evidence-based communications
animal performs species-specific behaviors with diversity between staff members, and to recommend that improve-
and frequency seen in the wild, especially those that the ments be considered, followed by a welfare reevaluation
animal is highly motivated to perform. The caregivers are after changes have been implemented. The AZA defines
often the most familiar with the animal’s behaviors and may animal welfare as an animal’s collective physical, mental,
observe when abnormal behaviors are present, for example, and emotional state over a period of time, which is measured
in terms of excessive activity or lack of activity, abnormal on a continuum from good to poor.25 For an institution to
vocalizations, excessive grooming or self-mutilation, or even be AZA accredited, it is required for that institution to
the development of a stereotypy.12 “develop and implement a clear and transparent process for
Abnormal repetitive behaviors, those without an obvious identifying, communicating, and addressing animal welfare
function, are called stereotypies, and are often behaviors concerns” in a timely manner without retribution to the
used to assess welfare. Stereotypical behavior has been staff member or volunteer who raises them.26 A committee
theorized to be the response of an animal to the pres- or other process must be developed to address any concerns
ence of abnormal stimuli or lack of stimuli in the captive regarding animal welfare within the institution. The com-
environment.19 There are situations in which a high level of mittee or process is supplementary to the normal chain of
abnormal behavior in an animal is associated with enhanced command within a zoo’s animal care department to ensure
coping abilities. A literature survey found that across species that the process is not influenced by personal motives or
and environments where data were provided, almost 68% conflicts. If concerns are not being addressed within the
of the situations that caused or increased stereotypies also standard chain of command, a welfare committee provides
decreased welfare, but also it warned that stereotypies were an option for matters to be addressed further and with
linked to good or neutral welfare nearly as often as poor and anonymity.26
therefore should not be used as a sole index of welfare.20 There is no definitive formula on how to initiate and
Assessment of mental welfare has also been referred to develop an animal welfare committee or process that is
as an indication of the subjective state of the animal. In standardized and comprehensive for every institution;
most situations, caregivers are unable to directly assess the however, the committee or process should include the fol-
thoughts and feelings of an animal and therefore must rely lowing elements: clear communication of the process to
on indirect indicators.12 Evaluation of an animal’s mental staff and volunteers; easy access to the committee by all
state may be the most abstract and anthropomorphic part staff and volunteers; staff with the experience and authority
of the welfare assessment because it requires observation necessary to evaluate submitted observations and implement
and evaluation of the animals’ demeanor or emotion. To any necessary changes; and timely feedback to the person
add to the challenge, there are few objective measures of submitting the observation.25,26 An animal welfare commit-
positive emotions in animals despite widespread physiologic tee within a zoo serves in an advisory and nonregulatory
and behavioral measures of negative emotions.21 Indications role to guide curatorial and executive-level staff in matters
of a positive affective state may include facial expressions, of animal welfare. The committee promotes scientific and
paw licks, tongue protrusions, and vocalizations, such as evidence-based, proactive metrics to develop best practices
purring in cats or chirping in rats. Unfortunately, these for animal care and to respond to welfare concerns raised
are not markers that can be easily extrapolated across all by staff and volunteers to hold the institution accountable
species.22 Multiple factors including appropriate husbandry for the provision of consistent optimal welfare.1 In some
and environment allow for the expression of natural behav- zoos, the animal welfare committee serves multiple roles,
iors and satisfactory mental well-being. Enrichment is one including reviewing research proposals and biomaterials
method of compensating for compromised conditions in requests, similar to an IACUC, and serves as a forum for
captivity.12 Enrichment programs try to provide opportuni- welfare discussions.
ties for animals to express behaviors driven by the positive The zoo animal welfare committee functions best when a
emotional systems of seeking, play, and caring, and decrease small group of members is selected for their leadership skills,
activation of the fear, rage, or panic systems.23,24 and includes staff with current or previous responsibilities
for animal welfare, care, and health across departments.
Animal Welfare Committees in Zoos The make-up of the committee varies among institutions,
but typically includes core internal members consisting of
With animal health and welfare as a top priority for zoos, a combination of executive level staff members, curators,
formation of an animal welfare committee within a zoo keepers, veterinarians, researchers, facilities managers, and
is beneficial to the institution on many levels. One of other department representatives. Veterinarians should
CHAPTER 12 Overview of Animal Welfare in Zoos 69
always be included in the welfare committee given their Once received, the chair of the animal welfare committee
expertise in medicine, biosecurity, zoonoses management, will acknowledge the concern and designate committee
and postmortem evaluation.1 Additional internal members members to investigate the complaint through direct
consisting of animal care and other department represen- observations, conversations with involved staff members,
tatives may also be included. Furthermore, zoo animal and other necessary means. If a concern cannot be rectified
welfare committees may include one or more community through initial evaluations, an ad hoc animal welfare com-
members.2 External members may consist of a combina- mittee meeting is initiated to investigate further. Following
tion of nonemployees who have interest or expertise in the investigation, and once a consensus is reached on an
animal care and welfare and may include board members, effective and realistic solution, recommendations will be
research associates, conservation partners, or local experts made to the executive level staff and curators overseeing the
from related fields. The committee is led by a chairperson, animal in question. It is the responsibility of the executive-
usually nominated by an executive level staff member, who level staff and curators to put the recommendations into
is responsible for committee organization, documentation, action and to continually work toward a zoo culture
and delegation. Other appointed or nominated positions, that always considers animal welfare in decision-making
such as vice-chair, may also exist, based on the institutional processes.27 When a consensus is not able to be reached by
structure. Membership of the committee may be permanent the committee, a zoo’s executive director or equivalent may
or on a rotational basis. Meetings are generally held quar- be consulted to make a final recommendation. Each step in
terly, though subcommittee or task force meetings and ad this process should be thoroughly documented, addressed
hoc meetings to address welfare concerns may be held more in a timely manner, and communicated to the initial person
frequently as needed. filing the complaint. Moreover, animal welfare committees
One of the most important functions of the welfare may also consider annual reports or updates to all zoo
committee is to respond to welfare concerns through a staff to maintain transparency, although anonymity of the
formal complaint driven system that encourages responsible complainants must be maintained, and potentially sensitive
reporting.27 If a welfare concern is not adequately addressed details tempered for a general audience. Additional guidance
through the standard chain of command within an animal on how best to establish an anonymous reporting system
care department, it may be raised to the welfare commit- may be found in the Animal Welfare Act Regulations.28
tee either through direct communication with one of the As zoos develop their own welfare committees and
committee members or through an anonymous reporting processes, there are many resources and examples available
phone line, email account, or online submission form. The (Table 12.3). In addition to individual institutions having
system should prompt the reporter to justify the concern their own welfare committees, larger management organi-
and offer potential solutions that are evidence based.27 zations also have welfare committees. The AZA’s welfare
TABLE
12.3 Resources for Assessment of Zoo Animal Welfare and Animal Welfare Committee Development
committee mission is to promote good welfare for animals Veterinarian’s Role in Zoo Animal Welfare
in AZA-accredited zoos, by assisting member institutions in
identifying and applying best practices in animal welfare, Veterinarians are knowledgeable animal care professionals,
and through promoting advances in animal science. It who have advanced medical training, scientific understand-
achieves its mission by promoting a common understand- ing, and communication abilities to be animal welfare
ing of animal welfare in the zoo and aquarium community; advocates and leaders within zoos.34–36 Moreover, veterinar-
assisting zoos and aquariums in identifying and applying ians may work under a legal mandate to have institutional
best practices in animal welfare; encouraging the develop- authority to oversee animal care and welfare, as with the
ment of research projects and assessment tools to advance Animal Welfare Act enacted by the US Department of Agri-
and monitor animal welfare; educating and engaging AZA culture.28 It is important to have veterinarians contribute to
zoos in applying assessment tools; and understanding and welfare assessments and participate in welfare committees,
influencing public perception about animal welfare in AZA in addition to assessing life-long medical needs, guiding
zoos.25,26 The AZA also offers support for and review of hospice and end-of-life decisions, and evaluating and inter-
animal welfare considerations for in situ, ex situ, and field preting postmortem information (see Chapter 15). It is
conservation work.29 In the United States, there are centers the responsibility of the veterinarian to implement existing
that serve to perform zoo animal welfare research and to welfare standards and strive for continued improvements
disseminate the findings.30,31 These are supported by AZA that are informed by medicine, behavior, ecology, and
and work to identify general needs and support progress ethics.37,38 The veterinarian is committed to the role as the
across North American zoos and aquariums. EAZA has animal’s advocate but may also have obligations to the insti-
an animal welfare training officer and has developed an tution, caregivers, peers within the profession, the public,
animal welfare working group, both of which are intended and himself or herself. The veterinarian may face challenges
to support the initiatives of all EAZA members aimed at when these commitments are conflicting. On a larger scale,
reaching high standards of animal welfare.32 On a global zoological veterinarians should be represented as stakehold-
scale, WAZA has developed an Animal Welfare Strategy to ers in the development of policies aimed at addressing
provide guidance on how to establish and maintain accept- welfare and should contribute to professional organization
able animal welfare standards and related best practices. welfare committees and working groups.25,34,39,40
It provides 58 recommendations in nine strategic areas, Focused education in animal welfare is expanding through
including: animal welfare and its assessment; monitoring and integration into veterinary curriculum and opportunities
management of animal welfare; environmental enrichment; for continued education in animal welfare exists across
exhibit design, breeding programs, and collection planning; all taxa.38,41–43 The American College of Animal Welfare
conservation welfare; animal welfare research; partnerships (ACAW) is a specialty organization focused on offering
in animal welfare; and engagement and interaction with training in animal welfare and board certification follow-
visitors.1 These resources should be consulted, along with ing either a formalized training program or experiential
fellow institutions, when developing a new animal welfare pathway.44,45 The Australian College of Veterinary Scientists
committee or process. has an Animal Welfare Chapter that offers training and cer-
tification.46 The Royal College of Veterinary Surgeons rec-
Animal Welfare Departments ognizes animal welfare science, ethics, and law as a specialty
subject area and offers credentials in the form of diplomas
In addition to animal welfare committees, some zoos em and certificates.47 The European College on Animal Welfare
ployee animal welfare directors within an animal welfare and Behavioral Medicine offers a subspecialty in Animal
department or institute to oversee animal welfare manage- Welfare Science, Ethics, and Law.48
ment, and to serve as a resource for animal care staff within
the zoo and for the greater zoo community. This role varies Acknowledgments
greatly between institutions but focuses on maintaining high
standards of animal welfare, as defined by the institution. The authors thank their colleagues Donald L. Janssen and
These departments may include oversight of animal train- Sharon Joseph for their contributions to this chapter.
ing, behavioral husbandry, enrichment, research review, and
technical support. Moreover, these departments may also References
perform routine welfare assessments for animals in conjunc-
1. World Association of Zoo and Aquariums: Caring for Wildlife,
tion with the curatorial and veterinary staff, review special
The World Zoo and Aquarium Animal Welfare Strategy 2015.
events hosted by the zoo and the impact on animal welfare, Available at: www.waza.org/en/site/home. (Accessed 9 June
and organize and conduct prospective studies. Behavioral 2017).
research through this department is a tool that can measure 2. Kagan R, Carter S, Allard S: A universal animal welfare frame-
welfare outputs, and the results aid in the development of work for zoos, J Appl Anim Welf Sci 18:S1–S10, 2015.
a database for making evidence-based decisions about 3. Maple TL: Toward a science of welfare for animals in the zoo, J
animal care.27 Appl Anim Welf Sci 10:63–70, 2007.
CHAPTER 12 Overview of Animal Welfare in Zoos 71
4. Hone D: Why zoos are good. Available at: www.davehone.co.uk. 25. Association of Zoos and Aquariums Web site: Animal Welfare
(Accessed 9 June 2017). Committee. Available at: www.aza.org/animal_welfare_
5. Kagan R, Veasey J: Challenges of zoo animal welfare. In Kleiman committee. (Accessed 9 June 2017).
DG, Thompson KV, Baer CK, editors: Wild mammals in captiv- 26. Association of Zoos and Aquariums Web site: The Accredita-
ity: principles and techniques for zoo management, ed 1, Chicago, tion Standards and Related Policies, 2017 Edition. Available at:
2010, University of Chicago Press, pp 11–21. www.aza.org/accreditation. (Accessed 9 June 2017).
6. Melfi VA: There are big gaps in our knowledge, and thus 27. Janssen DL, Miller L, Vicino G: Animal welfare and behavior:
approach, to zoo animal welfare: a case for evidence-based zoo Opportunities to thrive, in Proceedings. American Association of
animal management, Zoo Biol 28:574–588, 2009. Zoo Veterinarians annual conference 2014: 71–72.
7. Duncan IJH: Animal rights – animal welfare: a scientist’s assess- 28. United States Department of Agriculture. Animal Welfare
ment, Poult Sci 60:489–499, 1981. Act. Available at: www.nal.usda.gov/awic/animal-welfare-act.
8. Hill SP, Broom DM: Measuring zoo animal welfare: theory and (Accessed 9 June 2017).
practice, Zoo Biol 28:531–544, 2009. 29. Association of Zoos and Aquariums Animal Welfare Committee:
9. Koene P: Behavioral ecology of captive species: using behavioral Animal Welfare Considerations for Field Conservation Projects.
adaptations to assess and enhance welfare of nonhuman zoo Available at: www.aza.org/animal_welfare_committee, 2016.
animals, J Appl Anim Welf Sci 16:360–380, 2013. (Accessed 9 June 2017).
10. Mench JA, Kreger MD: Ethical and welfare issues associated 30. Chicago Zoological Society: Center for the Science of
with keeping wild mammals in captivity. In Kleiman D, Allen Animal Care and Welfare. Available at: www.czs.org/Centers
M, Thompson K, et al, editors: Wild mammals in captivity, ed 1, -of-Excellence/Center-for-Animal-Welfare. (Accessed 9 June
Chicago, 1996, University of Chicago Press, pp 5–15. 2017).
11. Siegford JM: Multidisciplinary approaches and assessment tech- 31. Detroit Zoological Society: Center for Zoo Animal Welfare.
niques to better understand and enhance zoo nonhuman animal Available at: www.detroitzoo.org/animals/center-for-zoo-animal-
welfare, J Appl Anim Welf Sci 16:300–318, 2013. welfare. (Accessed 9 June 2017).
12. Beaver BV, Bayne K: Animal welfare assessment considerations. 32. European Association of Zoos and Aquaria: Animal Welfare.
In Bayne K, Turner P, editors: Laboratory animal welfare, ed 1, Available at: www.eaza.net. (Accessed 9 June 2017).
Boston, 2014, Academic Press, pp 29–38. 33. American Veterinary Medical Association: Animal Welfare
13. Chicago Zoological Society: Animal Welfare Research: Welfare- Brochure. Available at: www.avma.org/KB/Resources/Reference/
Trak®. Available at: www.czs.org/Centers-of-Excellence/Center- AnimalWelfare. (Accessed 9 June 2017).
for-Animal-Welfare/Animal-Welfare-Research/WelfareTrak. 34. American Veterinary Medical Association: The veterinarian’s role in
(Accessed 9 June 2017). animal welfare April 2017. Available at: www.ebusiness.avma.org.
14. Cockram MS, Hughes BO: Health and diseases. In Appleby (Accessed 9 June 2017).
MC, Mench JA, Olsson IAS, et al, editors: Animal welfare, ed 2, 35. Ladewig J: The role of the veterinarian in animal welfare, Acta
Cambridge, 2011, CAB International, pp 120–137. Vet Scand 50:S5, 2008.
15. Blokhuis HJ: International cooperation in animal welfare: the 36. Berry J: Animal welfare….what is the veterinarian’s role? Can Vet
welfare quality project, Acta Vet Scand 50:S10, 2008. J 55:203–206, 2014.
16. Butterworth A, Mench JA, Wielebnowski N: Practical strategies 37. American Veterinary Medicine Association, Canadian Veterinary
to assess (and improve welfare. In Appleby MC, Mench JA, Medical Association, and Federation of Veterinarians of Europe:
Olsson IAS, et al, editors: Animal welfare, ed 2, Cambridge, The roles of veterinarians in ensuring good animal welfare. Avail-
2011, CAB International, pp 200–214. able at: www.fve.org. (Accessed 9 June 2017).
17. Moberg GP: Biological response to stress: implications for animal 38. World Veterinary Association: Policy on the role of the veterinar-
welfare. In Moberg GP, Mench JA, editors: The biology of animal ian in animal welfare. Available at: www.worldvet.org. (Accessed
stress: basic principles and implications for animal welfare, ed 1, 9 June 2017).
New York, 2000, CABI Publishing, pp 1–21. 39. American Association of Zoo Veterinarians: Legislation/animal
18. Keay JM, Singh J, Gaunt MC, et al: Fecal glucocorticoids and welfare committee information. Available at: www.aazv.org.
their metabolites as indicators of stress in various mammalian (Accessed 9 June 2017).
species: a literature review, J Zoo Wildl Med 37:234–244, 2006. 40. European Association of Zoo and Wildlife Veterinarians: Ethics
19. Garner JP, Meehan CL, Mench JA: Stereotypies in caged parrots, and welfare working group. Available at: www.eazwv.site-ym.com.
schizophrenia and autism: evidence for a common mechanism, (Accessed 9 June 2017).
Behav Brain Res 145:125–134, 2003. 41. Lord LK, Millman ST, Carbone L, et al: A model curriculum for
20. Mason GJ, Latham NR: Can’t stop, won’t stop: Is stereotypy a the study of animal welfare in colleges and schools of veterinary
reliable animal welfare indicator? Anim Welf 13:S57–S69, 2004. medicine, J Am Vet Med Assoc 250:632–640, 2017.
21. Mellor DJ: Enhancing animal welfare by creating opportunities 42. Association of Zoos and Aquariums: Animal welfare: Evidence-
for positive affective engagement, N Z Vet J 63:3–8, 2015. based management. Available at: www.aza.org. (Accessed 8 July
22. Yeates JW, Main DC: Assessment of positive welfare: a review, 2017).
Vet J 175:293–300, 2008. 43. San Diego Zoo Global: San Diego Zoo Global Academy: Animal
23. Grandin T, Johnson C: Animals make us human: creating the welfare. Available at: www.sdzglobalacademy.org. (Accessed 8
best life for animals, ed 1, New York, 2009, Houghton-Mifflin July 2017).
Harcourt. 44. American College of Animal Welfare Web site. Available at:
24. Morris CL, Grandin T, Irlbeck NA: Companion animals sym- www.acaw.org/acaw_home.html. (Accessed 9 June 2017).
posium: environmental enrichment for companion, exotic, and 45. Beaver BV: After the DVM: specialization in animal welfare, J
laboratory animals, J Anim Sci 89:4227–4238, 2011. Vet Med Educ 37:61–63, 2010.
72 SE C T I O N 2 Animal Welfare
46. Australia and New Zealand College of Veterinary Scientists: 53. British and Irish Association of Zoos and Aquariums: Animal
Animal Welfare Chapter. Available at: www.anzcvs.org.au. Welfare. Available at: www.biaza.org.uk. (Accessed 9 June 2017).
(Accessed 9 June 2017). 54. United States Department of Agriculture: Animal welfare infor-
47. Royal College of Veterinary Surgeons: Specialization in the mation center. Available at: www.nal.usda.gov/awic. (Accessed 9
veterinary profession: a consultation, October 2011. Available June 2017).
at: www.rcvs.org.uk. (Accessed 9 June 2017). 55. Zoo Aquarium Association: Professional Practices. Available at:
48. European College of Animal Welfare and Behavioural Medicine: www.zooaquarium.org.au. (Accessed 9 June 2017).
Animal Welfare. Available at: www.ecawbm.com. (Accessed 9 56. Zoological Association of America: Animal care and enclosure
June 2017). standards and related policies 2016 version. Available at:
49. World Organization for Animal Health: Animal Welfare. Avail- www.zaa.org. (Accessed 9 June 2017).
able at: www.oie.int. (Accessed 9 June 2017). 57. Vicino G, Miller L: From prevention of cruelty to optimizing
50. Farm Animal Welfare Council: Five Freedoms. Available welfare: Opportunities to thrive, in Proceedings. 34th Annual
at: www.webarchive.nationalarchives.gov.uk. (Accessed 9 June International Ethological conference 2015. Available at: www
2017). .behaviour-2015.m.asnevents.com.au. (Accessed 14 July 2017).
51. Yeates J: Patients. In Yeates J, editor: Animal welfare in veterinary
practice, ed 1, West Sussex, UK, 2013, Wiley-Blackwell, pp 1–32.
52. Mellor DJ, Beausoleil NJ: Extending the ‘Five Domains’ model
for animal welfare assessment to incorporate positive welfare
states, Anim Welf 24:241–253, 2015.
13
Stress and Animal Welfare—
Endocrinological Evaluation
CORINNE P. KOZLOWSKI
T
he selection of appropriate and objective measures to range of behavioral, health, and physiological responses is
monitor animal welfare is a priority for the zoo com- recommended to ensure that endocrine data are interpreted
munity. Glucocorticoid (GC) concentrations are a correctly.
commonly used physiologic metric for evaluating stress.
This chapter presents an overview of the stress response and Types of Samples Used for GC Assessment
methods for quantifying GCs from captive wildlife, with
considerations for sampling and data interpretation. GCs may be measured in a variety of sample types (blood,
saliva, urine, feces, hair, and feathers), depending on the
Overview of the Stress Response information required and the feasibility of collection,
particularly when repeated sampling over extended periods
Stress causes the hypothalamic-pituitary-adrenal (HPA) axis is necessary. Blood samples are difficult to obtain without
to mediate behavioral and physiologic changes that allow an inducing GC production, as handling, physical restraint,
individual to respond to a perceived challenge. Neurons in and collection procedures may elicit an adrenal response.7
the hypothalamus release corticotropin-releasing hormone Saliva samples are less invasive, but regular sample collec-
(CRH), which acts on receptors in the anterior pituitary tions may require substantial training. Furthermore, blood
and stimulates the release of adrenocorticotropic hormone and saliva samples are point samples that represent only
(ACTH) into the bloodstream. ACTH then causes the a short duration of time.7,8 Conclusions based on these
release of GCs (cortisol in most mammals and fish; corti- samples may be inaccurate as GCs are affected by circadian
costerone in amphibians, reptiles, birds, and rodents) from rhythms and exhibit daily fluctuations.
the adrenal cortex. Elevated production of GCs increases Monitoring adrenal activity through analysis of fecal or
energy availability and oxygen intake, enhances sensory urinary GCs is advantageous because an animal’s behavior
function and memory, and decreases pain perception. Blood and adrenal activity are not affected by the noninvasive
flow is reduced to organs not needed for movement, and collection process. Repeated sampling from individuals is
processes not immediately related to survival (e.g. digestion, possible, allowing for long-term monitoring of hormone
growth, immune function, and reproduction) are inhibited. changes. Samples may furthermore be collected from socially
These changes are adaptive in the short term but when housed animals without needing to separate individuals.
prolonged, they are associated with negative impacts on Fecal and urine samples are also not as strongly impacted
the neurologic, cardiovascular, immune, and reproductive by pulsatile secretion and circadian rhythms, and instead
systems. Fig. 13.1 illustrates GC patterns associated with represent an integrated measure of perceived stressors and
acute and chronic stress responses. resulting adrenal activity. However, knowledge of the lag-
In general, long-term increases in GC production may time between when a stressor is experienced and the appear-
be evidence of compromised welfare.1,2 However, adrenal ance of a signal is required for proper data interpretation.
responses also occur during beneficial behaviors that For feces, the lag-time depends on the intestinal transit time
require physical activity, including mating behavior and from the duodenum to the rectum and varies considerably
copulation,3 play sessions,4 and responses to environmental among species. Values typically range from 6–48 hours.9
enrichment.5 This may make it challenging to differentiate The lag-time is shorter for urine samples, generally ranging
between adaptive responses and those signaling genuine from 2–14 hours.9
stress. In addition, individuals of the same species may also Assessing GC production in hair and feathers, sample
differ in their responses as a result of differences in tempera- types not influenced by circadian rhythms, has the benefit
ment6 and previous experiences.1 Therefore, measuring a of easy collection and long-term stability. Free GCs diffuse
73
74 SE C T I O N 2 Animal Welfare
Glucocorticoid concentration
Glucocorticoid concentration
−10 0 10 20 30 −10 0 10 20 30
Time (days) Time (days)
A B
1200 1200
Fecal glucocorticoids (ng/g)
800 800
600 600
400 400
200 200
0 0
−10 0 10 20 30 −10 0 10 20 30
Time (days) Time (days)
C D
• Figure 13.1 Theoretical acute (A) and chronic (B) stress responses, as assessed by glucocorticoid
measures. Day 0 indicates the day a stressor was experienced. Acute stress is characterized by a
short-term increase in glucocorticoids above an individual’s baseline followed by a relatively rapid return
to baseline levels, whereas a chronic stress response occurs when concentrations remain elevated above
baseline for a significant length of time. The bottom panels present fecal glucocorticoid profiles from two
cheetahs, illustrating an acute and chronic stress response. One individual (C) experienced two temporary
increases in fecal glucocorticoids after arriving at a new institution. The second individual (D) experienced
a prolonged increase in fecal glucocorticoids following transfer to a new habitat. Note that glucocorticoid
concentrations in fecal material are typically more variable than blood measures, and proper interpretation
requires knowledge of individual baseline values.
into hair through the root, which is supplied by capillar- increase in plasma GC levels through injection of ACTH.12
ies, and through the highly vascularized base from which Samples collected before and after injection are analyzed
feathers originate. While hair grows slowly and is typically to confirm that increased GC concentrations are detected
collected to assess the impact of stressors over the course of following ACTH administration. An ACTH challenge test
weeks to months,10 feathers have bands that correspond to may not be possible with endangered or difficult species.
daily growth cycles that may be used to provide a record of A biological validation, serial sampling before and after
GC production over several days or weeks.11 However, little a known stressful event (e.g., a capture, medical proce-
is known about the contribution of glandular secretions dure, or transfer), may instead be performed in these
(sweat, sebum, or other scents) to the GC content of hair situations.1
and feathers, and an understanding of the growth rate is For any validation, it is important to consider the number
required for proper data interpretation. and ages of individuals tested and to include both sexes,
as baseline and peak concentrations of GCs may vary.13
Validation Procedures Females tend to have higher GC concentrations, possibly
as a result of differences in the affinity of steroid-binding
Proper validation involves determining whether sample globulins.14 Time of year and reproductive status should
storage and extraction procedures are appropriate, whether also be considered, as concentrations for some species are
the immunoassay antibodies may detect the metabolites higher during the breeding season,13 and values for females
in the samples (for feces and urine), and whether the may be affected by phase of the estrous cycle,15 pregnancy,
assay system detects biologically meaningful changes in or lactation.12 To minimize these effects, each animal should
GC production. A physiologic validation, also known as serve as its own control during a validation, as well as during
a challenge test, involves pharmacologically inducing an studies assessing individual welfare.
CHAPTER 13 Stress and Animal Welfare—Endocrinological Evaluation 75
O
ne currently accepted definition of behavior before (antecedents) and after the behavior (consequences)
states that “behavior is the internally coordinated is central to identifying potential etiologies and solutions.
responses (actions or inactions) of whole living To comprehensively describe these key antecedents and
organisms (individuals or groups) to internal and/or exter- consequences, two further steps are undertaken (Fig. 14.1).
nal stimuli, excluding responses more easily understood as Obtaining a thorough, systematic, and detailed history of
developmental changes.”1 Behavior is about what the animal the case is critically important as more than one factor is
does, how it acts (or not) in response to the environment. It generally involved. Additionally, a complete physical exami-
is not about how an animal is, because this would require nation of the animal should be performed, as underlying
inferences and anthropomorphic interpretations that might medical conditions might be overlooked when assessing
limit the capacity to efficiently solve behavior problems.2,3 As behavior problems in zoo animals.
every living organism constantly displays behaviors, issues
with behavior may arise in any species, from invertebrates to Applied Behavior Analysis
great apes, and include a large variety of situations that are
often very challenging for zoo veterinarians and managers. Every behavior has a function.3 Therefore a detailed descrip-
From inappropriate vocalization to self-mutilation, veteri- tion of the problematic behavior is essential. Ideally, the
narians may be consulted to help solve cases in which the problematic behavior, as well as bouts of normal behavior
animal is acting in an unexpected manner for the current from the affected individual, should be observed directly.
context. As for any other disorder, it is important to first Video recordings may also be used but do not always allow
establish a differential diagnosis list. Unexpected behaviors for optimal understanding of the entire situation. The
are often better considered as clinical signs rather than a problematic behavior needs to be described accurately using
specific problem entity. Just as weight loss or hematuria may the animal’s body language (i.e., the jaguar is licking his
have multiple causes, behaviors such as feather picking or tail, his pupils are dilated, he is growling, his body is stiff,
pacing may result from numerous different processes that etc.) and not labels that include many different interpreta-
require tailored treatment regimens. For example, pacing tions (i.e., the jaguar is stressed, he is aggressive, he does
associated with an inappropriate physical environment will not tolerate this situation, he is territorial, he is dominant,
be treated differently from pacing associated with gastric etc.). Many details of the animal’s behavior including
pain. Behaviors are the results of complex interactions of body language must be collected during this process.
genetic, epigenetic (factors affecting gene expression result- Using specific behavior descriptions instead of inferences
ing in phenotypes) and environmental factors.4 However, or human interpretations allows objective measurements
with a systematic approach, it is generally possible to list a and greatly improves communication among observers. In
number of potential causes or diagnoses for a given behav- several species, we may differentiate between behaviors of
ior, and solutions to appropriately address the problem. The “normal” animals that may be explained or understood
first important step is the observation and description of the within the context of the situation and “abnormal” behav-
animal’s behaviors in a variety of circumstances, including iors in “sick” animals that are more likely associated with a
the problematic situation. All behaviors of the animal medical condition such as an anxiety disorder. Description
involved, as well as those of conspecifics or other indi- of the behavior should include details on frequency and
viduals, are often useful to better understand the social duration as they may have a bearing on the diagnosis. For
and environmental contexts of the unexpected behavior. example, a cockatoo screaming once a day for 30 seconds
Once the problematic behavior has been identified, a is more likely to be normal than a cockatoo screaming 50
functional assessment aiming to identify what happens times a day for 10 minutes at a time. Magnitude or severity
76
CHAPTER 14 A Systematic Approach in Diagnosing Behavior Problems 77
History
Gender Rearing
Reproduction Play
Therapeutic trials
of the behavior should be well documented, as it will be well-framed functional assessment of the situation, provides
useful for both determining the diagnosis and monitoring essential data to understand the causes of the problematic
the case progression. A problematic behavior will rarely behavior and often offers a path to the most appropriate
disappear instantly with treatment, but might progressively intervention and therapeutic plan. To better identify distant
reduce in frequency, duration, and magnitude/severity. The and immediate functional antecedents as well as conse-
functional assessment of the problematic situation should quences that may be external or internal (such as pain),
also include analysis of what contributes to the behavior. taking a systematic history and performing a thorough
Antecedents include everything functionally related to the medical assessment are subsequent, crucial steps.
behavior that occurs before the problematic behavior, such
as the context, the potential triggers, and/or environmental Comprehensive Behavioral History
stimuli. Antecedents influence the likelihood that a spe-
cific behavior will occur, and their identification may help It is essential to gather basic signalment information,
determine the diagnosis.2 For example, when looking at a including gender and reproductive status, and a good
circling behavior in a male hippopotamus, the identified history to verify the accuracy of information in the medical
antecedents could be the female charging the male when records. Misidentification of the gender, whether by human
he walks toward the hay. In this example, working on the or laboratory error, may obviously lead to misinterpretation
antecedents by addressing the female’s behavior is part of of behaviors. As few methods have shown 100% accu-
the solution to resolving the male’s abnormal behavior. racy, the gender of nonsexually dimorphic species may be
Consequences include what happens immediately after the confirmed by combining methods appropriate to the age,
behavior and provide environmental feedback to the animal species, and condition of the animal. If contraception has
on what the behavior just performed actually reaped.3 Con- been used, details on the surgical technique or chemical
sequences influence whether the behavior will be repeated protocol administered should be provided. Vasectomy (and
or not in the future. By observing the consequences of a its potential failure) versus castration, or the use of contra-
specific behavior, one may generally predict if it will increase ceptive hormones versus immunocontraceptive agents, will
or decrease in the future. Continuing the aforementioned affect both sexual and nonsexual (i.e., social interaction,
hippopotamus example, the consequences of the female appetite, activity, etc.) behaviors in very different ways.5–7
behavior (charging the male) are that the male goes away Though still poorly understood in zoo animals, contra-
and that she acquires more hay to consume. These are ceptive agents may have an impact on mood disorders in
positive and desirable outcomes, making her behavior more humans and could be a concern in nonhuman primates.7,8
likely in the future. Therefore, behavior analysis, through a Use of temporary or permanent methods to prevent certain
78 SE C T I O N 2 Animal Welfare
species-typical behaviors, such as onychectomy, wing trim- stereotyped and self-directed behaviors in certain cases.48,49
ming, or amputation, may have behavioral effects that have Appropriate designs, for example with the use of visual
been well documented in pet species and should be noted barriers and hiding places, or by increasing size and envi-
in the history procurement.9 Previous medical and surgical ronmental complexity, may help decrease some problem
history should be collected to focus the physical examination behaviors.32,50–52 Water quality and the ratio of water to land
and assist in choosing any other appropriate diagnostics. should be reviewed for aquatic or semi-aquatic species.35
Reviewing keepers’ notes may deliver critical information, Subtle behavior changes, such as blepharospasm in case
as some minor, intermittent clinical signs, such as chronic of problems with sterilization systems in marine mammal
vomiting, might not have been considered as warranting pools or in aquaria, will often precede more obvious clinical
further investigation when initially detected. Wild versus signs.53 Exposure to natural weather and to appropriate
captive birth, rearing methods, and environment are criti- light type and photoperiod also decreases stress in several
cal factors influencing behavior throughout the life of the species.42,54 Time spent on-exhibit versus off-exhibit and
animal.10,11 Human-rearing, maternal separation, and early indoors versus outdoors may also shape problematic behav-
stress have been shown to negatively, but not always irre- iors.55,56 Daily routines, from keeper’s presence to feeding
versibly, impair the behavior of many different species, from schedule, should be evaluated in relation to the problem’s
great apes to starlings.12–24 Early environmental conditions occurrence. Fixed feeding and public presentation schedules
have also been shown to affect reptile and fish behavior.25,26 may provoke anticipatory behaviors.55,57–59 Food types and
As these history factors cannot be altered in adult animals, feeding methods in captivity might interfere with normal
knowledge of their existence is of little help to solve the behavior, whether related to the feeding location (grazing
observed behavior problem; however, their recognition on the ground versus eating from a high rock), quality
might enable caretaker staff to make more informed future (natural browse versus pellets, varying size of food items),
choices that will aid in the prevention of similar behavior quantity (one meal vs. foraging all day), and delivery (in a
problems. Social environment should be carefully evaluated bowl vs. on the ground).55,56,60 Feeding methods stimulat-
when assessing an animal for a behavioral issue, as the ing natural behaviors such as foraging are less likely to
identified individual may be the victim in a conflict situa- be associated with behavior problems.61,62 Other factors
tion. The animal could also be the aggressor, causing severe that need to be investigated when gathering information
group stress with subsequent behavioral abnormalities in about a behavior case include elimination, sleep, play,
other individuals. Although individual isolation in a social and exploratory behaviors. In many species, elimination
species may have severe consequences, group housing for behaviors provide useful information on social interactions,
solitary species, altering social structures to the needs of territorial use, and possible medical or anxiety disorders.
captive breeding programs, and retention of offspring with For example, urine-spraying has been linked to anxiety and
their parents for longer than natural periods may also secondary aggression in felids.29 Other forms of marking
lead to behavior problems.11,27–32 An appropriate social behaviors, such as rubbing their chest on various items, may
structure favors normal behavior in all species, including be relevant in certain situations.63 Sleep behavior, as well
aquatic species.33–35 Interactions with other species should as nocturnal activities, most commonly recorded through
be considered, even if only visual or olfactory.11 The effect video recordings using infrared light, may add a lot to the
of human presence, keepers or visitors, on zoo animals has history by showing if the abnormal behaviors are present
been studied in various situations, from routine presence or not at night and if the normal sleeping and resting
around the enclosure to active interactions. The influence time is disrupted or compatible with normal ranges.29,32,64
on the animal’s behavior, whether positive (e.g., through Exploratory behavior, an important part of the activity
training), neutral, or negative (e.g., by disrupting normal budget in some species, is often limited in captive settings.59
activities) is often very important as it contributes to either Play is another very important part of the normal repertoire
chronic stress or daily enrichment.36–43 Reproductive history, in many species and is considered by some authors as an
current status, and future breeding plans must be reviewed indicator of welfare. Changes in its normal pattern are
even when behavior surrounding reproduction is not the worth noting.33,43,65,66 Finally, reviewing the enrichment
main concern. Reproductive failure may be associated with protocol is important to have a complete picture of the
chronic environmental or social stressors and should be animal’s environment. Enrichment is designed to promote
considered as another clinical sign.11,35,44,45 Maternal neglect species-specific behaviors, should be encouraged for all
has been related to chronic stress (i.e., oxytocin inhibited classes of animals including invertebrates and fish, and has
by stress) and hand-rearing (e.g., in felids).21,22,46 Abnormal been shown to help decrease abnormal behaviors in some
behaviors affecting successful breeding are also seen in some situations.67–71
anxiety disorders.47 The current physical environment and By performing this attentive review of the animal’s
husbandry practices should be carefully reviewed due to history, several potential causative factors for the behav-
their primary importance in the development and main- ioral problem may already be identified. In tandem, ideas
tenance of abnormal behaviors in captive animals. Use of for solutions and improvement of the animal’s social and
more naturalistic and enriched habitats allows the animals physical environments will likely arise from this detailed
to display more species-typical behaviors and may reduce analysis.
CHAPTER 14 A Systematic Approach in Diagnosing Behavior Problems 79
Thorough Physical Examination not yet well described in the zoo animal literature, it is
likely that what is seen in domestic animals is applicable
With the exception of a few bird species, communication to similar species of wild animals. Some behaviors may
between animals and their caretakers is nonverbal. Hence, seem unrelated to a specific disease but are in fact the
only a careful observation of the animal’s behavior may help manner in which the animal reacts to pain, even if the
us detect the presence of physical discomfort or mental function of the “abnormal” behavior remains unclear.72
distress. As veterinarians, the primary role when address- Examples of behaviors possibly associated with pain include
ing behavior problems is to eliminate all potential medical growling, biting, lunging, scratching, licking, feather or
causes for the problematic behavior. Studies attempting to hair picking, pacing, and vocalizing.2,72 So-called stereotypic
find a treatment for a group of behavior problems such as and compulsive behaviors must also be investigated for
feather picking in parrots or pacing in cats usually fail at signs of underlying medical conditions, as many cases
finding a solution that works for every case. One of the have been successfully treated once a medical diagnosis
reasons is that the diagnoses underlying these behaviors was established.
and the function of the behavior for each individual may Behavior is likewise influenced by genetic and epigenetic
vary. When a bornavirus infection is present and the bird aspects through the expression of different genes at the
picks at its feathers possibly as a sign of discomfort, no neuronal level. Interaction of hormones and neurotrans-
environmental enrichment or antidepressant medication mitters is very complex, and as with any other organ,
will help decrease the behavior. Some behaviors are well- malfunction, whether from birth or acquired later, may
recognized signs of pain or underlying disease, but others occur. Psychiatry in zoo animals is still in its infancy and
are less commonly recognized (Table 14.1). For instance, mainly extrapolates knowledge and experience from human
fly biting behaviors and licking of surfaces have been and domestic animal medicine. Psychiatric diagnoses are
associated with gastrointestinal diseases in some dogs.72 controversial in most species and will likely remain so due
Treatment of the underlying medical condition resulted in to the inherent complexity of measuring the parameters
resolution of the abnormal behaviors in these dogs. Though involved in “mental” (neurochemical) diseases. However, in
veterinary medicine, we may refer to anxiety disorders with
some imperfect, but objective, assessment tools. Anxiety
in veterinary medicine is defined as the anticipation of
TABLE Examples of Behaviors Compatible With a future threat or danger (real or imaginary). While fear
14.1 Physical or Mental Diseases
is appropriate when confronted with real dangers, anxiety
Behaviors Systems Affected becomes a disease if the perceived threat is imaginary and
if it impairs the expression of normal behaviors and adap-
Abnormal elimination Urin/Neuro/Endocrino/GI/
(rubbing urine on body, Psych
tive responses.73 Animals with neurotransmitter disorders
etc.) (anxiety or other “mental” diseases) may behave in what
we often interpret as aberrant ways: performing painful
Aggressive behaviors Pain/Neuro/Endocrino/
(growling, lunging, etc.) Psych
or apparently pointless behaviors (i.e., self-mutilations) or
attacking individuals that had no prior interaction with them
Circling/pacing/increased Pain/Neuro/Repro/GI/ (i.e., redirected aggression). According to human psychiatry,
motor activity Psych
it is possible that these animals perceive a distorted reality
Flank/tail/leg sucking or GI/Dermato/Neuro that impacts their behavior. In parallel, fear and anxiety
licking significantly interfere with learning processes in the brain,
Fly biting/head extensions GI/Neuro/Ophthalmo which explains why abnormally fearful animals may not be
Hair plucking/feather picking Pain/GI/Dermato/Neuro/
easily trained, despite possibly normal to superior cogni-
Psych tive skills. Though a rare occurrence, recognizing “mental”
diseases in zoo animals may be challenging. These patients
Pica GI/Neuro
might require an appropriate medication to help balance
Prolonged piloerection Psych their brain neurochemistry and permit them to learn and
Restlessness Pain/Endocrino/Neuro/GI/ function again in a way similar to their conspecifics. Clinical
Psych signs suggestive of a “mental” illness such as anxiety disor-
Self-mutilations Pain/Dermato/Neuro/Psych
ders in zoo animals might include: startling even at routine
noises, not being able to habituate to new objects or new
Star gazing Neuro/GI/Pain/Psych situations, prolonged or frequent piloerection, incapacity to
Dermato: skin sensitivity, allergies, parasites, etc.; Endocrino: hypo- relax and sleep, prolonged recovery after stress, redirected
and hyperthyroidism, diabetes, Cushing, etc.; GI: gastrointestinal, IBD: aggression, etc.73 Normal arousal and vigilance levels highly
irritable bowel disease, dental diseases, pancreatitis, etc.; Neuro: vary across species, so comparing an individual to its group
congenital, cognitive dysfunction, neoplasia, etc.; Psych: Anxiety disor-
ders, compulsive disorders, etc.; Urin: idiopathic cystitis, infections, etc.; will help identify if the behavior is atypical.
Repro: uterine neoplasia, mastitis, ovarian cysts, etc. Finally, the information gathered through the three steps
described (applied behavior analysis, history, and physical
80 SE C T I O N 2 Animal Welfare
examination) must be analyzed simultaneously as many 4. Robinson GE: Beyond nature and nurture, Science 304:397–399,
elements are intrinsically related. Stress from the environ- 2004.
ment may cause gastric ulcers with secondary vomiting and 5. Patton ML, Jöchle W, Penfold LM: Review of contraception in
ungulate species, Zoo Biol 26:311–326, 2007.
growling at conspecifics. Treating both the gastric ulcers
6. De Nys HM, Bertschinger HJ, Turkstra JA, et al: Vaccination
and addressing the environmental concerns is more likely against GnRH may suppress aggressive behaviour and musth in
to resolve the abnormal aggressive behaviors than treating African elephant (Loxodonta africana) bulls: a pilot study, J S Afr
one or the other clinical sign. In addition, social status may Vet Assoc 81:8–15, 2010.
affect significant changes in the type of immune response in 7. Gray ME, Cameron EZ: Does contraceptive treatment in wildlife
nonhuman primates.74 Communication between the brain result in side effects? A review of quantitative and anecdotal
and neurons of the gastrointestinal tract has now been well- evidence, Reproduction 139:45–55, 2010.
described in humans, explaining the long-time observed 8. Skovlund CW, Mørch LS, Kessing LV, et al: Association of
association between mental and gastrointestinal diseases.75 hormonal contraception with depression, JAMA Psychiatry
From an observer’s vantage, behavior is simply the result of 73:1154–1159, 2016.
the animal interfacing with its environment. Behavior is in 9. Gerard AF, Larson M, Baldwin CJ, et al: Telephone survey to
investigate relationships between onychectomy or onychec-
fact the consequence of a complex and interconnected web
tomy technique and house soiling in cats, J Am Vet Med Assoc
of genes, their level of expression, and resulting physiol- 249:638–643, 2016.
ogy, affecting both perception and actions, in a constantly 10. Mason G: Are wild-born animals protected from stereotypy
changing environment. Regardless of the approach (ecology, when placed in captivity? In Mason GJ, Rushen J, editors:
genomics, etc.), animal behavior remains a fascinating topic Stereotypic animal behaviour: fundamentals and applications to
and needs further objective, nonanthropomorphic studies. welfare, Cambridge, 2006, CABI, p 196.
There are always individuals behaving differently from the 11. Lindburg DG, Fitch Snyder H: Use of behavior to evaluate repro-
majority. These differences may help define which behav- ductive problems in captive mammals, Zoo Biol 13:433–445,
iors are considered “inappropriate” or “abnormal” for a 1994.
species. Through a systematic approach to these behavior 12. Jacobson SL, Ross SR, Bloomsmith MA: Characterizing abnor-
“problems,” we will acquire a better understanding of the mal behavior in a large population of zoo-housed chimpanzees:
prevalence and potential influencing factors, Peer J 4:e2225,
underlying genetic, epigenetic, and environmental factors
2016.
that contribute to an animal exhibiting unexpected behav- 13. Martin JE: Early life experiences: activity levels and abnormal
iors. Obtaining a final diagnosis is not always feasible, but behaviours in resocialised chimpanzees, Anim Welf 11:419–436,
as we identify the potential origins of the problem, we will 2002.
also find viable solutions. Behavior problems are challeng- 14. Harlow HF, Harlow MK: The effects of rearing conditions on
ing for all including the affected animals, their caregivers, behavior, Bull Menninger Clin 26:213–224, 1962.
conspecifics (i.e., in cases of redirected aggression), and 15. Fox RA, Millam JR: Novelty and individual differences influ-
management dealing with visitor complaints. Addressing ence neophobia in orange-winged Amazon parrots (Amazona
behavior problems with an inclusive approach, in coopera- amazonica), Appl Anim Behav Sci 104:107–115, 2007.
tion with the veterinary team, keepers, and managers, is 16. Myers SA, Millam JR, Roudybush TE, et al: Reproductive
therefore an important step in improving animal welfare success of hand-reared vs. parent-reared cockatiels (Nymphicus
hollandicus), Auk 105:536–542, 1988.
in zoo animals.
17. Schmid R, Doherr MG, Steiger A: The influence of the breeding
method on the behaviour of adult African grey parrots (Psittacus
Acknowledgments erithacus), Appl Anim Behav Sci 98:293–307, 2006.
18. Korosi A, Naninck EFG, Oomen CA, et al: Early-life stress
I would like to acknowledge Leigh Clayton, Shannon Ferrell, mediated modulation of adult neurogenesis and behavior, Behav
Diane Frank, Susan Friedman, and Stéphane Lair for their Brain Res 227:400–409, 2012.
continuous guidance throughout my behavior learning 19. Meder A: Effects of hand-rearing on the behavioral development
experience and their thorough review of this chapter. of infant and juvenile gorillas (Gorilla g. gorilla), Dev Psychobiol
22:357–376, 1989.
20. Feenders G, Bateson M: Hand rearing affects emotional responses
References but not basic cognitive performance in European starlings, Anim
Behav 86:127–138, 2013.
1. Levitis DA, Lidicker WZ, Jr, Freund G: Behavioural biolo- 21. Kelling AS, Bashaw MJ, Bloomsmith MA, et al: Socialization of
gists do not agree on what constitutes behaviour, Anim Behav a single hand-reared tiger cub, J Appl Anim Welf Sci 16:47–63,
78:103–110, 2009. 2013.
2. van Zeeland Y, Friedman SG, Bergman L: Behavior. In Speer B, 22. Mellen JD: Effects of early rearing experience on subsequent
editor: Current therapy in avian medicine and surgery, St. Louis, adult sexual behavior using domestic cats (Felis catus) as a model
2015, Elsevier Health Sciences, pp 177–251. for exotic small felids, Zoo Biol 11:17–32, 1992.
3. Friedman SG, Edling TM: Concepts in behavior, section I: 23. Snyder RJ, Zhang AJ, Zhang ZH, et al: Behavioral and develop-
the natural science of behavior. In Harrison GJ, Lightfoot TL, mental consequences of early rearing experience for captive giant
editors: Clinical avian medicine, Palm Beach, 2006, Spix Publish- pandas (Ailuropoda melanoleuca), J Comp Psychol 117:235–245,
ing, pp 46–59. 2003.
CHAPTER 14 A Systematic Approach in Diagnosing Behavior Problems 81
24. Utt AC, Harvey NC, Hayes WK, et al: The effects of rearing 45. Zhang G, Swaisgood RR, Zhang H: Evaluation of behavioral
method on social behaviors of mentored, captive-reared juvenile factors influencing reproductive success and failure in captive
California condors, Zoo Biol 27:1–18, 2008. giant pandas, Zoo Biol 23:15–31, 2004.
25. Alberts AC: Behavioral considerations of headstarting as a con- 46. Mason GJ, Veasey JS: How should the psychological well-being of
servation strategy for endangered Caribbean rock iguanas, Appl zoo elephants be objectively investigated?, Zoo Biol 29:237–255,
Anim Behav Sci 102:380–391, 2007. 2010.
26. Salvanes AGV, Moberg O, Braithwaite VA: Effects of early 47. Holman HJ: Combination with fluoxetine and alprazolam to
experience on group behaviour in fish, Anim Behav 74:805–811, control anxiety in a hand-reared snow leopard (Uncia uncia), J
2007. Zoo Wildl Med 47:900–902, 2016.
27. Kraemer GW, Schmidt DE, Ebert MH: The behavioral neuro- 48. Clarke AS, Juno CJ, Maple TL: Behavioral effects of a change in
biology of self-injurious behavior in Rhesus Monkeys, Ann N Y the physical environment: a pilot study of captive chimpanzees,
Acad Sci 836:12–38, 1997. Zoo Biol 1:371–380, 1982.
28. Leeds A, Boyer D, Ross SR, et al: The effects of group type and 49. Fàbregas MC, Guillén Salazar F, Garcés Narro C: Do naturalistic
young silverbacks on wounding rates in western lowland gorilla enclosures provide suitable environments for zoo animals?, Zoo
(Gorilla gorilla gorilla) groups in North American zoos, Zoo Biol Biol 31:362–373, 2012.
34:296–304, 2015. 50. Bashaw MJ, Kelling AS, Bloomsmith MA, et al: Environmental
29. Miller A, Leighty KA, Bettinger TL: Behavioral analysis of tiger effects on the behavior of zoo-housed lions and tigers, with a case
night housing practices, Zoo Biol 32:189–194, 2013. study on the effects of a visual barrier on pacing, J Appl Anim
30. Waples KA, Gales NJ: Evaluating and minimising social stress in Welf Sci 10:95–109, 2007.
the care of captive bottlenose dolphins (Tursiops aduncus), Zoo 51. Chang TR, Forthman DL, Maple TL: Comparison of confined
Biol 21:5–26, 2002. mandrill (Mandrillus sphinx) behavior in traditional and “ecologi-
31. Kanghae H, Thongprajukaew K, Jatupornpitukchat S, et al: cally representative” exhibits, Zoo Biol 18:163–176, 1999.
Optimal-rearing density for head-starting green turtles (Chelonia 52. Quirke T, O’Riordan RM, Zuur A: Factors influencing the preva-
mydas Linnaeus, 1758), Zoo Biol 35:454–461, 2016. lence of stereotypical behaviour in captive cheetahs (Acinonyx
32. Montaudouin S, Pape GL: Comparison between 28 zoological jubatus), Appl Anim Behav Sci 142:189–197, 2012.
parks: stereotypic and social behaviours of captive brown bears 53. Colitz CMH, Saville WJA, Renner MS, et al: Risk factors
(Ursus arctos), Appl Anim Behav Sci 92:129–141, 2005. associated with cataracts and lens luxations in captive pinnipeds
33. Guarino S, Hill HM, Sigman J: Development of sociality and in the United States and the Bahamas, J Am Vet Med Assoc
emergence of independence in a killer whale (Orcinus orca) calf 237:429–436, 2010.
from birth to 36 months, Zoo Biol 35:2016. [Epub ahead of 54. Carvalho TB, Mendonça FZ, Costa-Ferreira RS, et al: The effect
print]. of increased light intensity on the aggressive behavior of the
34. Desjardins JK, Hofmann HA, Fernald RD: Social context influ- Nile tilapia, Oreochromis niloticus (Teleostei: Cichlidae), Zoologia
ences aggressive and courtship behavior in a Cichlid fish, PLoS (Curitiba) 30:125–129, 2013.
ONE 7:e32781–e32785, 2012. 55. Hasenjager MJ, Bergl RA: Environmental conditions associated
35. Marshall AR, Deere NJ, Little HA, et al: Husbandry and enclo- with repetitive behavior in a group of African elephants, Zoo Biol
sure influences on penguin behavior and conservation breeding, 34:201–210, 2015.
Zoo Biol 35:385–397, 2016. 56. Orban DA, Siegford JM, Snider RJ: Effects of guest feeding
36. Hosey GR: How does the zoo environment affect the behaviour programs on captive giraffe behavior, Zoo Biol 35:157–166,
of captive primates?, Appl Anim Behav Sci 90:107–129, 2005. 2016.
37. Thompson VD: Behavioral response of 12 ungulate species in 57. Jensen A-LM, Delfour F, Carter T: Anticipatory behavior in
captivity to the presence of humans, Zoo Biol 8:275–297, 1989. captive bottlenose dolphins (Tursiops truncatus): a preliminary
38. Fernandez EJ, Tamborski MA, Pickens SR, et al: Animal–visitor study, Zoo Biol 32:436–444, 2013.
interactions in the modern zoo: conflicts and interventions, Appl 58. Gilbert-Norton LB, Leaver LA, Shivik JA: The effect of randomly
Anim Behav Sci 120:1–8, 2009. altering the time and location of feeding on the behaviour of
39. Melfi V: Is training zoo animals enriching?, Appl Anim Behav Sci captive coyotes (Canis latrans), Appl Anim Behav Sci 120:179–185,
147:299–305, 2013. 2009.
40. Hosey G, Melfi V: Human–animal bonds between zoo profes- 59. Quirke T, O’Riordan R: An investigation into the prevalence of
sionals and the animals in their care, Zoo Biol 31:13–26, 2012. exploratory behavior in captive cheetahs (Acinonyx jubatus), Zoo
41. Kratochvil H, Schwammer H: Reducing acoustic disturbances Biol 34:130–138, 2015.
by aquarium visitors, Zoo Biol 16:349–353, 1997. 60. Rozek JC, Danner LM, Stucky PA, et al: Over-sized pellets
42. Ozella L, Anfossi L, Di Nardo F, et al: Effect of weather condi- naturalize foraging time of captive Orange-winged Amazon
tions and presence of visitors on adrenocortical activity in captive parrots (Amazona amazonica), Appl Anim Behav Sci 125:80–87,
African penguins (Spheniscus demersus), Gen Comp Endocrinol 2010.
242:49–58, 2017. 61. Jenny S, Schmid H: Effect of feeding boxes on the behavior of
43. Neto MP, Silveira M, Santos Dos ME: Training bottlenose stereotyping Amur tigers (Panthera tigris altaica) in the Zurich
dolphins to overcome avoidance of environmental enrichment Zoo, Zurich, Switzerland, Zoo Biol 21:573–584, 2002.
objects in order to stimulate play activities, Zoo Biol 35:210–215, 62. van Zeeland YRA, Schoemaker NJ, Ravesteijn MM, et al:
2016. Efficacy of foraging enrichments to increase foraging time in
44. Pimm RH, Dutton C, O’Handley S, et al: Assessment of the Grey parrots (Psittacus erithacus erithacus), Appl Anim Behav Sci
reproductive status of female veiled chameleons (Chamaeleo 149:87–102, 2013.
calyptratus) using hormonal, behavioural and physical traits, Zoo 63. Gaspari MK, Crockett CM: The role of scent marking in Lemur
Biol 34:20–32, 2015. catta agonistic behavior, Zoo Biol 3:123–132, 1984.
82 SE C T I O N 2 Animal Welfare
64. Wilson ML, Bashaw MJ, Fountain K, et al: Nocturnal behavior 70. Clark JA, Haseley A, Van Genderen G, et al: Increasing breeding
in a group of female African elephants, Zoo Biol 25:173–186, behaviors in a captive colony of Northern Bald Ibis through
2006. conspecific acoustic enrichment, Zoo Biol 31:71–81, 2012.
65. Vicino GA, Marcacci ES: Intensity of play behavior as a potential 71. Corcoran M: Environmental enrichment for aquatic animals, Vet
measure of welfare: a novel method for quantifying the integrated Clin North Am Exot Anim Pract 18:305–321, 2015.
intensity of behavior in African elephants, Zoo Biol 34:492–496, 72. Frank D: Recognizing behavioral signs of pain and disease: a
2015. guide for practitioners, Vet Clin North Am Small Anim Pract
66. Burghardt GM, Ward B, Rosscoe R: Problem of reptile play: 44:507–524, 2014.
environmental enrichment and play behavior in a captive Nile 73. Frank D: Distinguishing behavioral disorders from medical
soft-shelled turtle, Trionyx triunguis, Zoo Biol 15:223–238, 1996. disorders. In Ettinger SJ, Feldman EC, Côté E, editors: Textbook
67. Carlstead K, Shepherdson D: Effects of environmental enrich- of veterinary internal medicine, St. Louis, 2016, Elsevier Health
ment on reproduction, Zoo Biol 13:447–458, 1994. Sciences, pp 46–49.
68. Shyne A: Meta-analytic review of the effects of enrichment on 74. Snyder-Mackler N, Sanz J, Kohn JN, et al: Social status alters
stereotypic behavior in zoo mammals, Zoo Biol 25:317–337, immune regulation and response to infection in macaques,
2006. Science 354:1041–1045, 2016.
69. Näslund J, Johnsson JI: Environmental enrichment for fish in 75. Lyte M: Microbial endocrinology in the microbiome-gut-brain
captive environments: effects of physical structures and substrates, axis: how bacterial production and utilization of neurochemicals
Fish Fish 17:1–30, 2016. influence behavior, PLoS Pathog 9:e1003726, 2013.
15
Quality-of-Life Assessment
and End-of-Life Planning
for Geriatric Zoo Animals
LARRY VOGELNEST AND JESSICA J. TALBOT
83
84 SE C T I O N 2 Animal Welfare
B
• Figure 15.1 Advanced degenerative joint disease with complete loss of articular cartilage, eburna-
tion, extensive osteophytosis, enthesopathy, and bone cyst formation: (A) Kodiak bears (Ursus arctos
middendorffi) aged 28.4 years (left) and 31.2 years (right) at time of euthanasia; and (B) Komodo Dragon
(Varanus komodoensis) aged 32.8 years at time of euthanasia. (Photo credits: Taronga Zoo.)
TABLE
15.1 Age-Related Health Conditions in Zoo Animals by Body System
A B
• Figure 15.2 Radiographs (A) and necropsy images (B) of a northern giraffe (Giraffa camelopardalis),
aged 26 years at time of euthanasia, showing advanced periodontal dental disease with attrition.
TABLE
15.2 Suggested Roles for Key Stakeholders Providing Geriatric Zoo Animal End-of-Life Care
will form emotional attachments with animals in their Identification of aged animals in a zoo population is
care.20 Empathy and compassion for staff and provision of crucial to the process of quality-of-life assessment for these
support and counseling are important considerations.21 Zoo animals. Zoos should develop a methodology and database
managers and occupational health workers must also rec- to identify animals approaching or beyond an average or
ognize the emotional and psychological effect on veterinary “expected longevity” for the species. “Expected longevity”
teams caring for sick animals and performing euthanasia for a species may be calculated as the age at which 90%
and implement mechanisms to manage compassion fatigue. of a species’ population within regional species studbooks
This may include professional training on the recognition had died.5 Reported maximum longevities of zoo animals
of signs of compassion fatigue and promoting self-care should be interpreted with caution as these are generally
techniques.22,23 It is the responsibility of zoo veterinarians “outliers” and do not accurately reflect “normal” longevity
to educate other stakeholders on age-associated health for a species. Additionally, age-related changes are likely to
conditions, pain recognition, and discomfort, and to assess start well before an animal approaches maximum longevity.
the need for euthanasia to prevent and relieve suffering.6,24 The use of longevity records for a species in the wild is of
Communicating effectively about degenerative and chronic little use in identifying aged zoo animals as these are often
disease early in the end-of-life planning process helps avoid considerably less than longevity of the same species in zoos.
debate and prepares zoo staff for an animal’s euthanasia The stage in an animal’s life at which quality-of-life
when warranted.25 For iconic species or individuals, media assessment commences should be guided by certain trig-
team involvement in the process may help prepare and gers. One could argue that this process should start from
inform the wider community of the death of a zoo animal.26 Text continued on p. 90
86 SE C T I O N 2 Animal Welfare
• Figure 15.3 Example of a quality-of-life or aged animal assessment tool. The criteria are scored based
24. Downing R: Pain management for veterinary palliative care 30. Delaski KM, Ramsay E, Gamble KC: Retrospective analysis of
and hospice patients, Vet Clin North Am Small Anim Pract mortality in the North American captive Red Panda (Ailurus
41:531–550, 2011. fulgens) population, 1992–2012, J Zoo Wildl Med 46:779–788,
25. Irwin SA, Mausbach BT, Koo D, et al: Association between 2016.
hospice care and psychological outcomes in Alzheimer’s spousal 31. Lamglait B, Trunet E, Leclerc A: Retrospective study of mortality
caregivers, J Palliat Med 16:1450–1454, 2013. of captive African wild dogs (Lycaon pictus) in a French zoo
26. Wiedner E: Geriatric large carnivore medicine made easy, in (1974–2013), J Zoo Aquar Res 3:47–51, 2015.
Proceedings. NAVC Conference 2016; 1580–1582. 32. Fuller G, Lukas KE, Kuhar C, et al: A retrospective review
27. Chicago Zoological Society: WelfareTrak®. Available at: https:// of mortality in lorises and pottos in North American zoos,
www.welfaretrak.org. (Accessed 30 December 2016). 1980–2010, Endanger Species Res 23:205–217, 2014.
28. Lowenstine LJ, McManamon R, Terio KA: Comparative pathol- 33. Holz PH, Little PB: Degenerative leukoencephalopathy and
ogy of aging great apes: bonobos, chimpanzees, gorillas and myelopathy in dasyurids, J Wildl Dis 31:509–513, 1995.
orangutans, Vet Pathol 53:250–276, 2016. 34. Bunting EM, Garner MM, Abou-Mahdi N, et al: Proliferative
29. Kolmstetter C, Munson L, Ramsay EC: Degenerative spinal thyroid lesions and hyperthyroidism in captive fishers (Martes
disease in large felids, J Zoo Wildl Med 31:15–19, 2000. pennanti), J Zoo Wildl Med 41:296–308, 2010.
SECTION 3
Conservation Medicine
16 Evaluating Camel Health in Kenya—An Example of
Conservation Medicine in Action, 93
92
16
Evaluating Camel Health
in Kenya—An Example of
Conservation Medicine in Action
SHARON L. DEEM
93
94 SE C T I O N 3 Conservation Medicine
in the changing environment has declined; however, this of comparative medicine and discoveries in all life forms;
may also indicate three points of concern when viewed in and (6) demonstrating the importance of nature for human
a conservation medicine framework. First, with just one health.17–20 This chapter presents a conservation medicine
camel milk pasteurization plant in the country, Kenyans program focusing on camel health in Kenya, which dem-
have little access to pasteurized milk. It has been estimated onstrates roles that zoological veterinarians play to ensure
that 10% of Kenya’s 40 million people drink unpasteurized healthy animals and healthy people.
camel milk.11 Because raw camel milk is a possible trans-
mission route for many microorganisms, the consumption Project Design
of unpasteurized camel milk in Kenya may pose a high
public health cost in the country.11–15 Secondly, camels are In 2012, the Saint Louis Zoo Institute for Conservation
browsers and have great ability to cover large areas across Medicine (ICM) was invited to spearhead camel health
the landscape, which may lead to competition for food studies at the Mpala Research Centre (MRC) in Laiki-
with sympatric wildlife.16 Lastly, there remains a shortage pia County, Kenya (Fig. 16.1). The invitation was from
of veterinary education for camel health and a lack of colleagues working at MRC and Novus International,
veterinary support for camel production in the country. As an animal nutrition company with headquarters near St.
a relatively new large-scale production livestock species in Louis, MO. After researchers embarked on camel nutrition
Kenya, and as a species often viewed as most closely linked studies, they soon realized that little data existed on the
with marginalized people, veterinary care and biosecurity health status of, and disease risks for, camels in Kenya. (This
for camels in Kenya significantly lags behind that for more was just before Middle East Respiratory Syndrome [MERS-
traditional livestock (e.g., cattle, sheep, goats). This lack of CoV] was on the international stage with the first detection
care has potentially devastating implications for productiv- of MERS-CoV in a 25-year-old student in Jordan.21 See
ity losses related to disease-associated morbidity and mortal- also chapter in this volume: An Overview of Middle East
ity, as well as an increase in disease transmission between Respiratory Syndrome in the Middle East.) We at the Saint
camels, other livestock, wildlife, and humans. All these Louis Zoo immediately accepted this opportunity because
factors and challenges demand a conservation medicine the ICM had just launched as a new Zoo department, and
approach. one of the Zoo’s WildCare Institute conservation centers
had been focusing efforts on wildlife conservation in the
Horn of Africa for many years prior to this time.
What Is a Conservation Medicine With minimal research, it quickly became evident that
Approach and How Do Zoos Fit Within It? a camel conservation medicine program was important
because (1) camels had become the “new cow” in Kenya;
Although a number of definitions have been provided for (2) many diseases of camels may be transmitted to humans,
conservation medicine, one unifying theme may be that livestock, and wildlife; and (3) the region of Kenya where
it is a strategy that strives to expand transdisciplinary col- MRC is located has experienced the largest increase in
laborations and communications to improve the health camel numbers while still holding the highest densities of
of humans, animals, and the environment.17 Today, with wildlife in the country, although these numbers are falling
the push for AZA-accredited zoos to dedicate 3% of their significantly as humans and livestock move across the
revenue to conservation, the time is right for zoos to be landscape.9,22
leaders in the this initiative. This should be an easy fit as a To get the program underway, we took a multistep
core objective of zoo conservation is often to ensure healthy approach that included (1) conducting research to better
wildlife populations and ecosystems, without compromis- understand the species and region; (2) reaching out to
ing the health of humans. Thus, the conservation mission of international camel experts and livestock/wildlife and
accredited zoos fits perfectly within a conservation medicine conservation researchers in Kenya; and (3) securing funds
framework. and people to make the program a success (Box 16.1). First
Zoological institutions have an opportunity to play many was the task of gathering literature on dromedary camel
roles within conservation medicine programs and we may health and diseases, and to consider these in a conservation
be the advocates that help to ensure species’ conservation medicine framework. We focused on the diseases that were
is considered within these programs for improving human, of most concern for camel productivity and human health,
animal (e.g., domestic and wild), and environmental as well as diseases transmissible across the camel-livestock-
health.17–19 Briefly, these roles may include (1) providing human-wildlife interface. Dromedary camels may harbor
healthcare for zoological species, thus ensuring sustain- agents with zoonotic potential (e.g., Coxiella burnetii,
ability of biodiversity; (2) conducting studies on diseases Brucella spp., Toxoplasma gondii, Rift Valley fever, anthrax)
of conservation concern; (3) understanding diseases in zoo and/or those that may be transmitted only among camels,
wildlife as sentinels for emerging diseases of humans and other livestock and wildlife (e.g., blue tongue, bovine diar-
animals in surrounding areas; (4) performing surveillance of rhea virus, Trypanosoma evansi).23–29 It was also clear that
diseases in wild animals at the interface of wildlife, domestic some of these diseases (e.g., Brucella spp.) are difficult to
animals, and humans; (5) making contributions to the fields diagnose in camels.
CHAPTER 16 Evaluating Camel Health in Kenya—An Example of Conservation Medicine in Action 95
• Figure 16.1 Map of Laikipia County, Kenya. (From Browne AS, Fèvre EM, Kinnaird M, et al: Sero-
survey of Coxiella burnetii (Q fever) in Dromedary Camels (Camelus dromedarius) in Laikipia County,
Kenya. Zoonoses Public Health 2017. doi:10.1111/zph.12337. http://onlinelibrary.wiley.com/doi/10.1111/
zph.12337/full#zph12337-fig-0001.)
After gaining an appreciation for the issues at hand, we Laikipia County), herders started to appreciate that with
needed to determine how to fund the program and what the movement of animals comes the movement of all their
partnerships with other conservation medicine practitioners micro- and macrobiota.
we could develop. Seed money from Novus International We also focused much of our work on C. burnetii,
helped get the project started. Subsequently, we were able because the original literature review indicated that Q fever
to secure funds from internal grants at the Saint Louis Zoo, was an emerging infectious disease (EID) in Kenya, and that
foundations, private donors, and governmental agencies. during recent years it had been causing significant disease in
Collaborations quickly grew in the first year and we con- Kenyans.34 However, with little known on the epidemiology
tinue to have new organizations working with us on camel of the disease in Kenya, we elected to explore the possible
health issues. These collaborations became increasingly easy role of camels. Our studies have demonstrated a high sero-
to develop following the discovery of MERS-CoV, and the prevalence to Q fever in camels, and the potential role they
role that camels have in the epidemiology of the disease.21 may serve as reservoirs of the bacteria, with implications
for cross-species transmission between livestock, sympatric
wildlife, and humans.35,36
Program Outcomes to Date and Rather serendipitously and shortly after we had started
Future Plans working with camels, the emergence of MERS-CoV in the
Middle East, with spread to other regions of the world,
Now entering the fifth year of this camel program in North- was a catalyst for the development of this conservation
ern Kenya, we have produced a number of scientific and medicine program. The use of samples we had collected and
layperson-friendly products to help with camel production, bio-banked in the initial years and just prior to MERS-CoV
while minimizing human and wildlife health concerns. Early being on the world stage provided an important source of
in the study we developed a camel herd health protocol that data for understanding this significant EID in the region.37
was shared with local camel herders in the region.32 Also, Lastly, a large part of our camel program in Kenya
after the first field season and with the documentation has been the training of next generation conservation
that mastitis was indeed causing high production losses, medicine practitioners through involvement in a real-life
we conducted a study to better understand the risk factors public health and wildlife conservation program.19 This has
associated with mastitis prevalence.33 Reports were also included students from Kenya, the United Kingdom, and
shared with local camel herders to provide information on the United States. Through this program, we have trained
the health status of their animals based on complete blood DVM, MSc, and public health students and provided
counts, chemistry profiles, and exposure to a number or them experiences in which they gain an appreciation for
infectious and parasitic disease-causing agents. the disease risks associated with changing environmental
One example of an immediate improvement in camel conditions, protein sources for humans, and the inevitable
welfare and productivity was the diagnosis, using simple increase in interactions at the domestic animal/wildlife/
blood film evaluations, of T. evansi in a camel herd that human interface.
was experiencing high calf mortality and severe morbid-
ity in adults (Fig. 16.2). The herd owner had not been Future Work
using trypanocide for prevention because trypanosomiasis
was thought to not occur in the region. However, in this This program is ongoing and we continue to focus on all
increasingly interconnected world, with animal movements four core components. These include: (1) improvements in
(e.g., camels from northern Africa and the Middle East into camel husbandry, health, and production including plans
for a camel veterinary course at the University of Nairobi
College of Veterinary Medicine; (2) Q fever and MERS-CoV
epidemiologic studies with emphasis on the role of vectors
and use of slaughterhouses for surveillance, respectively;
(3) training of conservation medicine practitioners; and (4)
continued collaborations across institutions and disciplines.
Developed in 2012 and at a time that few people had
camel health on their radar, we now know of the importance
of camels in the epidemiology of MERS-CoV and other
EIDs (e.g., Q fever) in Kenya.35–37 Therefore there is an
increase in organizations and individuals that are working
to improve camel production in Kenya, and we continue
to expand partnerships within this program. Only through
scientific research, veterinary medical care and public policy
for camel production will we be able to mitigate the potential
• Figure 16.2 Adult camel with clinical signs of, and blood film risks to public health, and sympatric wildlife and livestock
confirmation for, Trypanosoma evansi. (Photo credit: Sharon L. Deem.) health, while advancing camel productivity in the region.
CHAPTER 16 Evaluating Camel Health in Kenya—An Example of Conservation Medicine in Action 97
We continue as one of the collaborators working to advance conflict resolution, food security), it is imperative that the
camel welfare, health, and productivity; all imperative to conservation of wildlife species be included in the equa-
help support human health and, as importantly, to be sure tion if we are to properly address human, animal, and
that this new form of climate change adaptation and food environmental health, resilience, and ultimately survival.
security does not lead to unchecked negative impacts on the
conservation of Kenya’s amazing wildlife.
References
1. FAO: The state of the world’s land and water resources for food and
Concluding Thoughts on Conservation agriculture (SOLAW) – Managing systems at risk, London, 2011,
Medicine Programs Food and Agriculture Organization of the United Nations, Rome
and Earthscan, p 308.
This work on camel health in Northern Kenya is one ex- 2. Delgado C, Rosegrant M, Steinfeld H, et al: Livestock to 2020:
ample of a transdisciplinary conservation medicine program the next food revolution, Outlook Agric 30:27–29, 2001.
initiated by a zoo conservation medicine department. The 3. Mahamat MB, Crump L, Tidjani A, et al: Food security, nutri-
program has already generated the data necessary for better tion and the One Health nexus. In Zinsstag J, Schelling E,
understanding diseases at the camel, livestock, wildlife, and Waltner-Toews D, et al, editors: One Health: the theory and
human interface, in the face of changing environments. Zoo practice of integrated health approaches, Boston, 2015, CABI, pp
272–282.
staff and veterinarians are often well-versed in this across-taxa
4. Watson EE, Kochore HH, Dabasso BH: Camels and climate
approach. Some may question why a zoological institution resilience: adaptation in Northern Kenya, Hum Ecol Interdiscip J
and zoological veterinarian took the lead on developing a 44:701–713, 2016.
program for a “domestic livestock” species. However, when 5. Rawlence B: City of thorns: nine lives in the world’s largest refugee
viewed at the intersection of camel, livestock, wildlife, camp. Picador. 400 pp.
and human health and disease concerns, this is the type 6. Food and Agriculture Organization of the United Nations:
of program zoos should increasingly embrace. Zoological FAOSTAT. Available at: http://faostat3.fao.org/download/Q/
veterinarians are first and foremost veterinarians; we have QA/E. (Accessed 16 Mar 2016).
an ability to partner with public health colleagues, and 7. Ogutu JO, Piepho H-P, Said MY, et al: Extreme wildlife declines
zoos are working to increase their “fence to field” reach to and concurrent increase in livestock numbers in Kenya: what are
work for the conservation of wildlife species, while ensuring the causes?, PLoS ONE 11:e0163249, 2016.
8. FAOSTAT. 2013. http://faostat3.fao.org. (Accessed 16 April
human livelihoods and health are not compromised.17,19 As
2015).
previously stated,38 one solution to disease and conservation 9. Kagunyu AW, Wanjohi J: Camel rearing replacing cattle produc-
at the wild-domestic animal interface is the implementation tion among the Borana community in Isiolo County of Northern
of a proactive approach—addressing po tential pathogen Kenya, as climate variability bites, Pastoralism 4:13, 2014.
transmission before a volatile problem occurs. This program 10. Bosire CK, Ogutu JO, Said MY, et al: Trends and spatial varia-
did just that. tion in water and land footprints of meat and milk production
Charles Darwin is reported to have said that the species systems in Kenya, Agric Ecosyst Environ 205:36–47, 2015.
that survive are not necessarily the strongest or the most 11. Kaindi DWM: Microbiological quality of camel milk along the
intelligent; rather they are the ones that are the most adapt- market chain and its correlation with food-borne illness among
able to change.39 This grassroots conservation medicine children and young adults in Isiolo, Kenya. Master thesis at
program, which began prior to camels being on the world the Department of Food Science, Nutrition and Technology,
University of Nairobi.
stage due to their role in the emergence of MERS-CoV,
12. Cerf O, Condron R: Coxiella burnetii and milk pasteurization:
best demonstrates how the ability to adapt and attend to an early application of the precautionary principle?, Epidemiol
concerns of the day are in everyone’s best interest. A proac- Infect 134:946–951, 2006.
tive approach to camel health, at a time that camel numbers 13. Hussein MF, Alshaikh MA, Al-Jumaah RS, et al: The Arabian
are growing across the Kenyan landscape, with potential camel (Camelus dromedarius) as a major reservoir of Q fever in
negative impacts for wildlife conservation, is one example of Saudi Arabia, Comp Clin Path 24:887–892, 2014.
how zoos may lead in the conservation medicine initiative 14. Kaindi DWM, Schelling E, Wangoh JM, et al: Risk factors for
to help address these 21st century challenges. We took an symptoms of gastrointestinal illness in rural town, Isiolo, Kenya,
opportunity that at first glance might not have seemed a Zoonoses Public Health 59:118–125, 2012.
“good fit” for a zoo conservation department but was shown 15. Osoro EM, Munyua P, Omulo S, et al: Strong association
very early in our work to be an excellent fit. between human and animal brucella seropositivity in a linked
study in Kenya, 2012–2013, Am J Trop Med Hyg 93:224–231,
While working on this chapter, security issues in north-
2015.
ern Kenya again escalated due to worsening drought with 16. O’Connor DA, Butt B, Foufopoulos JB: Foraging ecologies of
dire consequences for pastoralists, herders, and ranchers in giraffe (Giraffa camelopardalis reticulata) and camels (Camelus
the region. Food security, and simple human security, will dromedarius) in northern Kenya: effects of habitat structure and
worsen in the short term, if not also the long term. We will possibilities for competition?, Afr J Ecol 53:183–193, 2015.
have to be ready for these changes. As we strive for improve- 17. Deem SL: Conservation medicine to One Health: the role of
ments in human livelihoods (e.g., poverty alleviation, zoologic veterinarians. In Miller RE, Fowler ME, editors: Fowler’s
98 SE C T I O N 3 Conservation Medicine
zoo and wild animal medicine (vol 8). Saint Louis, 2015, Elsevier 30. Rich KM, Perry BD: The economic and poverty impacts of
Saunders, pp 698–703. animal diseases in developing countries: new roles, new demands
18. Deem SL, Dennis P: The role of zoos in One Health. In One for economics and epidemiology, Prev Vet Med 101:133–147,
Health Newsletter (vol 5). Gainesville, 2012, University of Florida 2011.
(Emerging Pathogens Institute) One Health Initiative, pp 4–7. 31. Seifu E, Tafesse B: Prevalence and etiology of mastitis in tra-
http://media.news.health.ufl.edu/misc/egh/One%20Health%20 ditionally managed camels (Camelus dromedarius) in selected
Newsletter/FDOH%20Newsletters/Vol5Issue4_Fall12.pdf. pastoral areas in eastern Ethiopia, EthiopVet J 14:103–114,
19. Robinette C, Saffran L, Ruple A, et al: Zoos and public health: a 2010.
partnership on the One Health frontier, One Health 3:1–4, 2017. 32. Browne AS, Deem SL: Herd Health Protocols for Dromedary
20. Sahrmann JM, Niedbalski A, Bradshaw L, et al: Changes in Camels (Camelus dromedarius) at Mpala Ranch and Research
human health parameters associated with a touch tank experience Centre, Laikipia County, 2012, Kenya.
at a zoological institution, Zoo Biol 35:4–13, 2016. 33. Budd LH, Kinnaird M, Smith W, et al: Predictors of milk
21. Kupferschmidt K: The camel connection, Sci 1422–1424, 2014. quality and mastitis prevalence in dromedary camels (Camelus
22. Kinnaird MF, O’Brien TG: Effects of private-land use, livestock dromedarius) in northern Kenya. Trop Anim Health Prod (in
management and human tolerance on diversity, distribution and preparation).
abundance of large African mammals, J Cons Biol 26:1026–1039, 34. Njeru J, Henning K, Pletz MW, et al: Febrile patients admitted
2012. to remote hospitals in Northeastern Kenya: seroprevelance, risk
23. Afzal M, Sakkir M: Survey of antibodies against various infec- factors and a clinical prediction tool for Q-Fever, BMC Infect Dis
tious disease agents in racing camels in Abu Dhabi, United Arab 16:244, 2016.
Emirates, Rev - Off Int Epizoot 13:787–792, 1994. 35. Browne AS, Fèvre EM, Kinnaird M, et al: Serosurvey of Coxiella
24. Al-Ani FK, Sharrif LA, Al-Rawashdeh OF, et al: Camel diseases burnetii (Q fever) in Dromedary Camels (Camelus dromedarius)
in Jordan, in Proceedings, Third Annu Meet Animal Prod Under in Laikipia County, Kenya, Zoonoses Public Health 2017. http://
Arid Cond 2:77–92, 1998. dx.doi.org/10.1111/zph.12337.
25. Davies FG, Koros J, Mbugua H: Rift Valley fever in Kenya: 36. DePuy W, Benka V, Massey A, et al: Q fever risk across a
the presence of antibody to the virus in camels (Camelus drom- dynamic, heterogeneous landscape in Central Kenya, Ecohealth
edarius), J Hyg (Lond ) 94:241–244, 1985. 11:429–433, 2014.
26. El-Harrak M, Martin-Folgar R, Llorente F, et al: Rift Valley and 37. Deem SL, Fevre EM, Kinnaird M, et al: Serological evidence of
West Nile virus antibodies in camels, North Africa, Emerg Infect MERS-CoV antibodies in dromedary camels (Camelus drom-
Dis 17:2372–2374, 2011. edarius) in Laikipia County, Kenya, PLoS ONE 10:e0140125,
27. Mustafa IE: Bacterial diseases of dromedaries and bactrian 2015.
camels, Rev - Off Int Epizoot 6:391–405, 1987. 38. Mazet JAK, Johnson CK: Approaching health problems at the
28. Rahimi E, Ameri M, Karim G, et al: Prevalence of Coxiella wildlife-domestic animal interface. In Miller RE, Fowler ME,
burnetii in bulk milk samples from dairy bovine, ovine, caprine, editors: Fowler’s zoo and wild animal medicine, current therapy
and camel herds in Iran as determined by polymerase chain (vol 7). Saint Louis, 2012, Elsevier Saunders, pp 153–160.
reaction, Foodborne Pathog Dis 8:307–310, 2011. 39. Darwin C: On the origin of species by means of natural selection:
29. The World Organisation for Animal Health (OIE): Infectious or the preservation of favoured races in the struggle for life, London,
diseases of interest for camelids, appendix IV. Report of the 1859, John Murray.
second meeting of the OIE Ad hoc Group on Diseases of Cam-
elids. Paris: OIE; 3–5 May 2.
17
Disease Risks to Native Wildlife From
Zoos and Aquariums
BRUCE RIDEOUT AND CATHERINE HADFIELD
99
100 SE C T I O N 3 Conservation Medicine
in a new population or species by providing a prolonged transmission, which means efficient transmission occurs
transmission opportunity.12 In other words, there is a lower even in very small, low-density populations. For patho-
risk that the pathogen would die out before a transmission gens with transmission by direct contact, the probability
opportunity presents itself. The long infectious period of of encountering a susceptible host in a small, low-density
M. bovis likely contributed to its successful invasion and population is low, and if exposure occurs, invasion might
establishment in East Africa after spillover from domestic fail because transmission cannot be sustained. However,
animals.13 with vector-borne pathogens, transmission depends on the
Pathogens with rapid rates of evolution have greater frequency of encounters between the vector and host, not
invasion potential because of their ability to rapidly adapt the host density, so invasion and transmission occur even
to novel hosts.14 The classic example is RNA viruses, which in very small, dispersed populations as long as the vector
have high mutation rates during replication, as well as the is abundant. This also means that vector-borne pathogens
potential for reassortment and recombination events, allow- have the potential to drive small populations to extinction,
ing them to rapidly alter their host range and virulence. because transmission efficiency is maintained regardless
With avian influenza viruses, simple point mutations can of host population size or density. A classic example of
result in a shift of host range, allowing a switch to transmis- this scenario is the introduction of avian malaria into
sion between mammalian hosts.15,16 Hawaii, which has driven a number of native forest birds
Vector-borne pathogens have inherent limitations in host to extinction.17,18
switching potential because of the lifecycle requirements Representative examples of wildlife pathogens with these
and feeding preferences of their vectors, but those with gen- characteristics are listed in Table 17.1.
eralist vectors can have high invasion potential for several In order for any pathogen to become established in a zoo
reasons. First, vectors act as short-term reservoirs, allowing or aquarium and have a subsequent spillover opportunity, it
a pathogen to persist for a period of time in the absence of must first gain access to the facility by passing successfully
susceptible hosts, which increases exposure opportunities. through the quarantine process, or by breaching established
Second, vector-borne pathogens have frequency-dependent biosecurity barriers. The pathogens that are most successful
TABLE Representative Wildlife Pathogens With Characteristics That Facilitate Host-Switching and That Have the
17.1 Potential for Population-Level Impacts
Long Rapid
Taxon Agent Generalist Reservoirs Infectiousness Evolution Vectors*
Birds Circoviruses X X X
Bornaviruses X X† X X
HPAI X X X
Plasmodium spp. X X X X
Mycoplasma gallisepticum X X X
West Nile virus X X X X X
Mammals Mycobacterium tuberculosis complex X X X
Morbilliviruses X X X
Sarcoptes scabei X X X
Parvoviruses X X
Mycobacterium paratuberculosis X X X
Malignant Catarrhal Fever viruses X X X
Treponeme-associated Hoof Disease X X X
Toxoplasma gondii X X X
Reptiles Ranaviruses X X X
Snake Fungal Disease X X† X
Amphibians Batrachochytrium spp. X X X
Ranaviruses X X X
Fish Gyrodactylus spp. X X X X*
Sea Lice (Caligus spp.) X X X
Koi Herpesvirus X X
Carp Edema (Pox) Virus X X
Iridoviruses X X X
in making it through the quarantine process are those that this case focused broadly on the risk of alien pathogen
have a long incubation period or a carrier state, or that spillover to native wildlife, and includes the development of
lack effective screening tests, which makes them difficult to a consensus statement on the level of risk that is acceptable.
detect, or those allowed through quarantine because they are This is followed by the identification of all potential hazards,
considered inconsequential in their native host. Pathogens primarily infectious agents in this case, and an analysis that
with these characteristics should be given particular atten- culminates in a ranking of agents based on the likelihood
tion in the risk analysis process described later. The ability that a spillover event would occur and a negative population-
of a pathogen to breach established biosecurity barriers is level impact would follow. It is important to recognize that
determined more by the care that went into the design of pathogen spillover is a multistep process, and the probability
the barriers and the effectiveness of the biosecurity practices needs to be weighed at every step. For example, the analysis
at the institution than by the characteristics of the pathogen. might consider the estimated probability (usually expressed
The general biosecurity practices, disease surveillance, qualitatively as low, medium, or high) that an agent of
and preventive medicine programs of the facility are a concern would be present, that the agent could escape the
foundational component of any effort to mitigate the risk facility by some avenue, that exposure to native wildlife
of disease spillover to native wildlife. However, the potential would occur, that the hosts would be susceptible, that a
consequences of a spillover event are great enough to warrant productive infection would occur, that the pathogen would
more targeted risk analysis and mitigation efforts. One become established in a population, and that a negative
efficient approach is to combine this targeted risk analysis population-level impact would follow. To the extent these
with any new biosecurity planning efforts being initiated, steps are independent, the probability of each step needs to
such as those resulting from the increased threat of HPAI be multiplied rather than added in order to establish the
and other foreign animal disease incursions. The all-hazards cumulative probability. When using qualitative estimates,
response plans being developed and promoted through the the key concept to remember is that multiplication results
Association of Zoos and Aquariums (https://zahp.aza.org/) in a reduction in cumulative probability. This can be seen
are a good model (see Chapter 9). The same types of bios- by arbitrarily assigning quantitative estimates at each step,
ecurity practices that will protect our collection animals and doing the calculation (e.g., assigning a 50% probability
from pathogen spillover from native wildlife will also help for each step in a seven-step process, as outlined earlier,
to mitigate the risk of spillover in the other direction. A yields a cumulative probability of [0.5 × 0.5 × 0.5 × 0.5
biosecurity audit conducted by government animal health × 0.5 × 0.5 × 0.5] = 0.0078, which is <1%). See Table
regulatory veterinarians is an excellent starting point. The 17.2 for an example of a cumulative probability analysis
audit will help identify gaps or weaknesses in existing facili- for desert tortoises. For those agents that have a cumula-
ties and biosecurity protocols, and the findings can provide tive probability of negative population impact that exceeds
foundational information for a subsequent comprehensive the established risk tolerance level, mitigation steps would
risk analysis. then be developed. A useful tool for conducting such a
risk analysis is the International Union for Conservation of
Risk Analysis Nature (IUCN) Manual of Procedures for Wildlife Disease
Risk Analysis,19 which is available as a free download from
The risk analysis (see Chapter 2) should ideally have two the IUCN website (www.iucn.org).
components. The first would be a traditional risk analysis The second component of the risk analysis would be
focusing on pathogens of concern that have a reasonable focused on biosecurity for specific transmission pathways
probability of being present in the zoo or aquarium. The rather than specific pathogens. This is important because it
emphasis should be on pathogens that would be alien to has the potential to mitigate the risk of spillover of pathogens
native wildlife, especially those with the high-risk charac- that we might not have considered, or that might not have
teristics outlined previously. The success of the risk analysis been discovered or characterized yet. The process would
hinges on assembling a multidisciplinary team with the be somewhat analogous to what food safety specialists call
expertise and institutional knowledge required to identify Hazard Analysis and Critical Control Points (HACCP).20,21
and evaluate all avenues of risk relevant to the institutional The idea with this approach would be to map out all of the
setting. The composition of the risk analysis team will vary potential avenues of pathogen escape from a facility and
depending on the nature of the institution but should then identify the critical points in each escape or transmis-
include those with expertise in husbandry, management, sion pathway where risk mitigation would be most effective.
nutrition, and facility infrastructure, in addition to those Particular attention should be paid to situations that result
with traditional animal health expertise. It can also be very in recurring exposure opportunities, such as areas of water
helpful to include stakeholders, such as financial decision runoff or discharge from enclosures, fecal compost piles,
makers and government agency representatives responsible food waste cleanup and disposal practices, fomites, and
for native wildlife management. Even if they are not active areas where wildlife or feral animals can come into close
participants in the risk analysis process, their input and proximity with collection animals or enclosures.
buy-in is often critical to the success of the effort. The Complicating these risk scenarios is the potential role zoos
risk analysis process begins with a problem definition, in play in altering wildlife disease prevalence and dynamics by
102 SE C T I O N 3 Conservation Medicine
TABLE
17.2 Example of Cumulative Risk Analysis for Mojave Desert Tortoises
Probability
Absence in Destination
In Source Population
Translocation Is Only
Negative Population
Exposure Avenue
Agent or Hazard
Cumulative Risk
Establishment
Comments
Population
Impact
Mycoplasma Very Low Low Very Very High D: High High D: A remote naïve population
agassizii High High High Low D: Low Medium may be at high cumulative
Low D: Low risk in a high-density
scenario and low risk in
low density
Mycoplasma Very Low Low Very Very High D: Medium High D: Low
testudineum High High High Low D: Very Low Low D: Very
Low
TeHV-2 High Low Low Very Very Low Very Low Most taxa have multiple
High High endemic herpesviruses
High D, High tortoise population density; Low D, low tortoise population density; TeHV-2, testudinid herpesvirus 2.
Excerpt from Rideout B, editor: Transmissible infections and desert tortoise translocation: a comprehensive disease risk analysis. Report to the US Fish and
Wildlife Service, 2015.
inadvertently subsidizing urban-adapted wildlife, such as • Maintain closed aquatic systems, or ensure that dis-
deer, raccoons, skunks, rodents, opossums, foxes, coyotes, charged water is effectively treated before it enters a
mallards, Canada geese, herons, egrets, and feral cats. Our watershed or other area with native species.
facilities foster higher densities of these species by providing • Keep animals destined for reintroduction completely
abundant food and water, reduced predator populations, separate from species from other geographic areas and
and complex environments that provide shelter. Because ideally maintain them as close as possible to native
of this, risk mitigation efforts should also incorporate habitat and release areas.
biosecurity practices that minimize the subsidization of • Do not bring species into close proximity that would
urban-adapted wildlife. not have opportunities to interact in the wild, especially
Examples of targeted risk mitigation efforts could include species from different continents or divergent geographic
the following: regions.
• Minimize the ability of urban-adapted wildlife to • Ensure that appropriate biosecurity barriers and practices
obtain animal or human food waste from enclosures or are in place to minimize host-switching opportunities by
trashcans. pathogens.
• Employ integrated pest management and vector control • Minimize the length of time food waste is left in enclo-
practices that keep pests and urban-adapted wildlife at sures, where it could attract native wildlife.
the lowest possible population levels. The fact that there are no published reports of direct
• Schedule a site visit with experts in wildlife population spillover of significant pathogens from zoos and aquariums
control, such as US Department of Agriculture’s Animal to native wildlife suggests that existing biosecurity and pre-
and Plant Inspection Service’s staff in the United States, ventative medicine programs in zoos and aquariums have
to get specific recommendations for methods to reduce been relatively effective. However, the introduction of dis-
urban wildlife populations in facilities. eases from nonzoo or aquarium sources to North American
• Eliminate mosquito and other pathogen vector breeding wildlife, such as West Nile Virus, illustrates the potential for
sites. adverse effects of disease introduction. These risks are real
• Ensure that perimeter fences and facility barriers and need to be addressed, particularly for reintroduction
minimize the transit of urban-adapted wildlife through programs. Examples of diseases introduced through rein-
facilities. troduction include B. dendrobatidis in Mallorcan midwife
• Minimize the ability of free-ranging aquatic birds to toads,22 and whirling disease in rainbow trout.23 Although
access water features that either have collection birds, or there have been no documented population-level impacts
that collect runoff or discharges from exhibits. on native wildlife from spillover in zoos, there are examples
CHAPTER 17 Disease Risks to Native Wildlife From Zoos and Aquariums 103
of pathogen exchange occurring, such as with Isospora spp. 12. André J-B, Day T: The effect of disease life history on the
coccidia in zoos and free-ranging passerines.24 Continued evolutionary emergence of novel pathogens, Proc Biol Sci
anthropogenic impacts at the wildlife–domestic animal– 272:1949–1956, 2005.
13. Cleaveland S, Mlengeya T, Kazwala RR, et al: Tuberculosis in
human interface will only increase the risk of significant
Tanzanian wildlife, J Wildl Dis 41:446–453, 2005.
pathogen spillover events in the future. 14. Duffy S, Shackelton LA, Holmes EC: Rates of evolutionary
change in viruses: patterns and determinants, Nat Rev Genet
References 9:267–276, 2008.
15. Schrauwen EJ, Fouchier RA: Host adaptation and transmission
1. Lymbery AJ, Morine M, Kanani HG, et al: Co-invaders: of influenza A viruses in mammals, Emerg Microbes Infect 3:e9,
the effects of alien parasites on native hosts, Int J Parasitol 2014. Available at: http://dx.doi.org/10.1038/emi.2014.9.
Parasites Wildl 3:171–177, 2014. Available at: http://dx.doi 16. Shi Y, Wu Y, Zhang W, et al: Enabling the “host jump”: struc-
.org/10.1016/j.ijppaw.2014.04.002. tural determinants of receptor-binding specificity in influenza
2. Hudson PJ, Dobson AP, Lafferty KD: Is a healthy ecosystem one A viruses, Nat Rev Microbiol 12:822–831, 2014. Available at:
that is rich in parasites?, Trends Ecol Evol (Amst) 21:381–385, http://www.nature.com/doifinder/10.1038/nrmicro3362.
2006. 17. Warner RE: The role of introduced diseases in the extinction
3. Brearley G, Rhodes J, Bradley A, et al: Wildlife disease prevalence of the endemic Hawaiian avifauna, Condor 70:101–120, 1968.
in human-modified landscapes, Biol Rev 88:427–442, 2013. Available at: http://www.jstor.org/stable/10.2307/1365954.
4. Bradley CA, Altizer S: Urbanization and the ecology of wildlife 18. Woodworth BL, Atkinson CT, Lapointe DA, et al: Host popula-
diseases, Trends Ecol Evol (Amst) 22:95–102, 2007. tion persistence in the face of introduced vector-borne diseases:
5. Rideout BA, Sainsbury AW, Hudson PJ: Which parasites should Hawaii amakihi and avian malaria, Proc Natl Acad Sci USA
we be most concerned about in wildlife translocations?, Ecohealth 102:1531–1536, 2005.
1–5, 2016. 19. Jakob-Hoff RM, MacDiarmid SC, Miller PS, et al: Manual of
6. Cleaveland S, Laurenson MK, Taylor LH: Diseases of humans procedures for wildlife disease risk analysis. 2014:1–163.
and their domestic mammals: pathogen characteristics, host 20. Khandke SS, Mayes T: HACCP implementation: a practical
range and the risk of emergence, Philos Trans R Soc Lond B Biol guide to the implementation of the HACCP plan, Food Control
Sci 356:991–999, 2001. 9:103–109, 1998. Available at: http://www.sciencedirect.com/
7. Cleaveland S, Haydon DT, Taylor L: Overviews of pathogen science/article/pii/S0956713597000650.
emergence: which pathogens emerge, when and why?, Curr Top 21. Hulebak KL, Schlosser W: Hazard analysis and critical control
Microbiol Immunol 315:85–111, 2007. point (HACCP) history and conceptual overview, Risk Anal
8. Tompkins DM, Draycott RAH, Hudson PJ: Field evidence 22:547–552, 2002.
for apparent competition mediated via the shared parasites of 22. Walker SF, Bosch J, James TY, et al: Invasive pathogens threaten
two gamebird species, Ecol Lett 3:10–14, 2000. Available at: species recovery programs, Curr Biol 18:853–854, 2008.
isi:000085367700004. 23. Viggers K, Lindenmayer D, Spratt D: The importance of disease
9. Kelly DW, Paterson RA, Townsend CR, et al: Parasite spillback: in reintroduction programmes, Wildl Res 20:687, 1993. Available
a neglected concept in invasion ecology?, Ecology 90:2047–2056, at: http://www.publish.csiro.au/?paper=WR9930687.
2009. 24. Schrenzel MD, Maalouf GA, Gaffney PM, et al: Molecular char-
10. McCallum H: Disease and the dynamics of extinction, Philos acterization of isosporoid coccidia (Isospora and Atoxoplasma
Trans R Soc B Biol Sci 367:2828–2839, 2012. spp.) in passerine birds, J Parasitol 91:635–647, 2005.
11. McCallum H, Dobson A: Detecting disease and parasite threats
to endangered species and ecosystems, Trends Ecol Evol (Amst)
10:190–194, 1995.
18
Feral Cat Dilemma
KERRIE ANNE T. LOYD AND SONIA MARIA HERNANDEZ
Brief Natural History of Cats contributing to the large numbers of homeless cats today.
Homeless domestic cats may be feral (stray, unfriendly,
In this chapter, we discuss the origin and abundance of often untamed and unsocialized) or stray, but somewhat
domestic cats, the issues surrounding cat overpopulation tame. Large feral populations of cats may have originated in
and loss of wildlife, and the stakeholders involved in the developed countries as more people migrated to cities from
controversy surrounding cat management. We also summa- the countrysides in the early 20th century. High-density
rize potential solutions to address the growing populations urban living and the struggle to make ends meet likely led to
of cats. difficulties keeping pets; this along with lack of sterilization
techniques and the prolific nature of cats led to large feral
Cats and Human Culture populations in cities by the mid 20th century.8 Cats may
have more than two litters of four or more kittens a year,9,10
Cats are believed to be domesticated from the wildcat in the and immigration into areas of sufficient food may be very
Near East approximately 10,000 years ago. There is evidence high.11 Baker et al.12 recorded cat densities of 229–523 cats/
for their first link with humans from Cyprus,1 and cats have km2 in an urban area of the United Kingdom, far higher
been found in drawings of ancient Egyptians where they were than native mesopredator densities (averaging 37 animals/
worshipped,2 some images displaying felines with collars. km2 for red foxes) (Vulpes vulpes). Others summarized cat
Domestication also occurred in early Chinese civilizations density observations and listed over 2000 cats/km2 in sites
due to cats’ useful ability to remove rodents.2 Domestic cats in urban Rome, Italy; Jerusalem, Israel; and Ainoshima,
were originally valued as predators of pests around grain Japan.13 Many Americans support or maintain neighbor-
and crop stores and revered as religious figures. Today, cats hood colonies of feral cats,14 and this is not uncommon in
are cherished as companion animals, welcomed into our developed nations around the world.
homes as family members, and are responsible for regular
Internet “sensations.” Annual surveys by the American Pet The Controversy
Products Association continue to report that cats are the
most popular pet in America and cats have surpassed dogs The number of feral domestic cats in the United States is
as the most popular companion animal in most of North unclear but is thought to be in the tens of millions.15,16
America and Europe.3 Humans often develop strong emo- Such high population estimates have implications for both
tional connections to pet cats, and there is some evidence wildlife and public health,17,18 and there is broad interest
that pet ownership contributes to short-term improvement from community groups, nonprofits, and management
in human health,4 though hypotheses about contributions agencies in reducing cat populations. Biologically effective,
to mental and physical health remain inconclusive overall.5 yet socially acceptable, management strategies for feral cats
Even if pet ownership may not contribute to a longer, are a matter of contention in the United States and in
healthier life, any cat owner will provide anecdotal support many developed countries abroad.19 Islands with imminent
for the happiness and entertainment provided by their conservation issues due to cats have used pathogens,
companion on a daily basis. poison baiting (Australia, currently), and other lethal
control methods. Historically, community management has
The Rise of Feral Cats involved capturing and either socializing for the purpose of
adoption or euthanizing unwanted feral cats at local shel-
Domestic cats have long been a common sight in urban and ters. Citing the lack of success by animal shelters to decrease
suburban areas throughout the world, and the number of cat populations, a second strategy growing in popularity,
cats is continuously growing. It is expected that a minority involves trapping cats, sterilizing them and releasing the cats
of cat-owners in the United States (25%–50%) confine back to the site of capture (Trap-Neuter-Release [TNR]).14
their pets to an indoor lifestyle,6,7 and this may be one factor This control method is considered more humane than
104
CHAPTER 18 Feral Cat Dilemma 105
some cat activists claim domestic cats are adapted to live tured Eastern chipmunk (Tamias striatus) to its residence. (National
outdoors and provide biologically inaccurate justifications Geographic Remote Imaging. Athens, GA, 2011.)
that cats “play an important role in balancing the local
ecosystem.”23 Wildlife advocates argue for cat removal from what and how much they hunt and whether it has real
the environment as cats are an invasive species detrimental implications on prey at the population level. Domestic
to wildlife.24 Lauber et al.25 found ethical judgments of cats are thought to pose a significant threat to the birds,
those supporting fertility control (TNR) for cats included herpetofauna, and small mammals that they prey upon (Fig.
concern over killing animals to satisfy human interests 18.1).27,28,29 Despite domestication occurring over thousands
and protection of the individual cats. In contrast, lethal of years, cats retain the instinct and skill of hunting. Cats
control is often advocated by people who believe fertility have been documented killing a prey item even while eating
control (TNR) works too slowly,25 or not at all. Support their favorite food,30 and Barratt31 reported the number of
for TNR has undoubtedly paralleled society’s overall move- prey that cats captured was not influenced by the number
ment toward animal welfare values orientations and no-kill of meals provided. Davis32 observed that domestic cats con-
shelters. It is worth noting that pro-cat groups spend large tinued to hunt rats and pigeons during periods of supple-
sums of money to support TNR and lobby local and state mental feeding and that feeding did not decrease hunting.
governments to change ordinances to allow TNR activities. Cats have now been implicated in 63 species extinctions
For example, PetSmart Charities donated over 20 million on islands33 but have also been found to have negative
dollars toward TNR activities.26 impacts on songbirds in noninsular environments.6,12 A
recent estimate of the broad impact of free-roaming cats
Cats Hunt—So What? on the wildlife of the United States suggests that up to 3.7
billion birds and 20.7 billion mammals fall prey to cats each
Ecologists and conservation biologists are most concerned year.34 Feral cats are responsible for a greater proportion of
with the direct impacts of high numbers of feral cats on the kills than pet cats (Fig. 18.2). This may be related to
populations of native wildlife but are also concerned about the increased amount of time feral cats spend outdoors. In
competition with native predators for food and the spread fact, we found a significantly higher proportion of stray
of zoonotic disease due to interspecific interactions. At the colony cats to exhibit hunting behavior compared to pet
center of the controversy is not that cats hunt, but rather cats studied in Georgia. Sterilization does not appear to
106 SE C T I O N 3 Conservation Medicine
affect a cat’s motivation to hunt.35 Although much effort resulted in a large percentage of deaths or euthanasia.44
is devoted to the quantification of how much wildlife cats Cat depredation was the foremost cause for admission
hunt, the results are unlikely to produce enough evidence of bats to rescue centers in Italy45 and for juvenile blue-
to convince many cat activists. The presence of millions tongue lizards (Tiliqua scincoides) in Sydney, Australia.46
of free-roaming cats (both feral and pets) is undeniable. Interactions with cats was the second greatest cause of
Wildlife faces many threats, some of which are difficult to small-mammal admissions to the Wildlife Center of Vir-
predict and prevent (e.g., emergent diseases). The impact ginia over a 10-year period and the second greatest cause
from invasive species is both predictable and preventable. of avian mortality at the center.47 We recently reported that
Urban and suburban ecosystems serve as habitat to cats contribute substantially to cases presented to wildlife
diverse mammals, reptiles, and amphibians as well as rehabilitation hospitals and even with extensive veterinary
resident and migratory songbirds.6 Suburban environments intervention, their potential for recovery and release was
(e.g., backyards, zoos, parks), contain fragmented islands very slim. We reviewed data collected from 82 wildlife
of natural habitats, surrounded by roads and development rehabilitation centers throughout North America during a
that act as barriers to wildlife movement and exert other 3.5-year period to determine common causes of admission
anthropogenic influences on the health of natural systems and found domestic pets to be responsible for 14% of
(pollution, sediment run-off, loss of plant food sources, bird admissions, the second most common identifiable cause
collisions with windows, etc.). Due to the decline of natural of wildlife injury. Greater numbers of birds than reptiles,
areas and the rapid expansion of developed areas,36 urban amphibians, or mammals were admitted for rehabilitation
and suburban habitats are critical to the future protection as a result of domestic cat attack, and 78% of these did
of biodiversity. Suburban backyard habitats may provide not survive. The majority of immature individual animals
valuable resources to native wildlife, but they may become of all species submitted because of cat attacks also died or
ecologic traps if they harbor non-native predators. had to be euthanized because of the severity of injuries.48
Wildlife rehabilitation data raises awareness of the prevent-
Animal Welfare—From Both Sides able suffering of individual animals injured by domestic
species. One argument proposed by cat activists about the
There is also a large group of stakeholders concerned with activities of feral cats is that cats have a right to express
the welfare of abandoned and feral cats. A few studies report their natural hunting behavior. The welfare of individual
higher disease prevalence among cats living in feral colonies cats receives extensive attention from advocates and many
than in owned cats.37,38 Feral cats are subject to environmen- members of the public, whereby the welfare of individual
tal extremes, and various sources of trauma and predation, wildlife usually remains unseen.16
all of which contribute to high mortality rates,9,37,39,40 and
relatively short life spans.40 In a study where we outfitted
cats with point-of-view cameras, we found free-roaming Public and Wildlife Health Concerns
cats engaged in daily risky behaviors, most commonly: Related to Feral Cats
crossing roads, interacting with other cats, drinking from
puddles in parking lots, exploring storm drain systems, and Cats are reservoirs for numerous pathogens of zoonotic
entering crawlspaces of buildings.41 The evidence of the concern, including Bartonella henselae, Salmonella spp.,
poor quality of life of feral cats has led one animal rights Toxocara cati, tapeworms, hookworms, and Sarcoptes
group, People for the Ethical Treatment of Animals (PETA), scabiei.49,50 Cats are the definitive host for Toxoplasma gondii,
to agree that euthanasia is the most humane management which has been increasingly linked to negative neurologic
option for homeless cats42; however, others claim feral cats, issues in humans, ranging from schizophrenia to cognitive
and especially well-managed cat colonies, may live health- function in children.51,52 By far, the most serious zoonosis
ful, relatively long lives (e.g., HSUS, Alley Cat Allies, Best of feral cats is rabies, and people exposed to the virus are
Friends Animal Society). These organizations support the often associated with feral cats (i.e., living near or caring for
maintenance of feral cat colonies over lethal control. colonies). Outside of cases due to bats, human exposure to
Whereby cat welfare is well advertised and supported, rabies is primarily associated with feral cats.53,54 One study
wildlife victims of cats do not receive as much attention, summarizing 10 years of data in South Carolina found that
yet personnel involved in wildlife rescue and rehabilitation feral cats were more likely to be rabid than stray dogs and the
regularly treat animals injured by domestic cats. Wildlife second most common species to expose humans to rabies
rehabilitation continues to gain popularity and National (behind foxes).55 Even though many cats in managed TNR
Wildlife Rehabilitators Association (NWRA) membership colonies receive a rabies vaccination when sterilized, they
has grown from approximately 200 to >1800 members do not receive boosters and are vulnerable to infection after
since 1984. The exact number of animals rehabilitated short-term protection, particularly from other terrestrial
is difficult to estimate, but during 2007 alone, NWRA rabies vector species (e.g., raccoons) (Procyon lotor) that tend
members reported treating over 100,000 animals.43 Cats to aggregate at feral cat feeding stations.56 Another serious,
were among the top anthropogenic causes for submission but more rare, concern for people interacting with feral cats
of injured animals to a wildlife clinic in Tennessee and including caregivers and veterinarians is plague (Yersinia
CHAPTER 18 Feral Cat Dilemma 107
pestis). This bacteria is endemic in the western United States, a happier, healthier life, as well as help protect wildlife. They
and cats are responsible for at least one infection every year.8 may provide a solution, particularly for managing smaller
Increasing urbanization and its associated high numbers of cat colonies, and this option should be explored by more
feral cats lead to further potential for increased contact and cites and nonprofit organizations. Cats could be trapped,
transmission of disease between cats and people.57 neutered, and then released only within the boundaries of
The transmission of pathogens across multiple hosts has an enclosure where they are safe from vehicles and predators
been documented in several locations at the urban-wildland and where wildlife outside the fence is protected. Enclosed
interface,58 and cats may serve as reservoirs for significant colonies would require property, enclosure materials, and
pathogens that affect other wildlife, such as Feline Leukemia structures—a large initial cost; however, many organizations
Virus (FeLV), which was transmitted from feral cats to the (e.g., Alley Cat Allies), corporations (e.g., PetSmart), and
endangered Florida panther (Puma concolor coryi).59 Since local county and city jurisdictions that donate millions of
the discovery that Toxoplasma gondii was responsible for dollars annually to TNR lobbying and education might
mortalities of sea otters (Enhydra lutris) in California,60 contribute to funding them. In addition, TNR programs
the scope and results of studies elucidating the detrimental already utilize dozens of dedicated volunteers64 who could
effects of this introduced pathogen into the marine ecosys- instead focus their efforts on care and adoption of cats
tem have broadened.61 at these facilities. Enclosed sanctuaries may also provide
unique opportunities for socializing the tamest of the cats
until they are adoptable. Currently, there are few examples
Management Alternatives to of such successful efforts (e.g., Chico Cat Coalition, Chico,
Trap-Neuter-Release or Euthanasia California; Blind Cat Rescue and Sanctuary, St. Pauls, North
Carolina). Before these enclosures can be promoted as an
Ultimately, feral cat numbers are unlikely to decline unless alternative solution, scientific evaluation of the feasibility of
multiple strategies are adopted in combination. At the core creating and maintaining sanctuaries is recommended. In
of that, and depending on the region, a vigorous trap and the end, if we do not come together to create a combination
removal program is likely needed. In addition, strongly of strategies that all groups can support, cat effects on
enforced, effective licensing, identification, and confine- biodiversity may become so apparent that it may lead to
ment laws are integral. Continued financial support for more extreme activities such as the baiting and poisoning
animal control agencies and shelters that are tasked to exhibited in Australia to protect threatened and endangered
remove, socialize, and adopt animals is also very impor- species.65
tant. Currently, US and European societies do not allow or
support feral dog populations and many propose the same
should be true for cats. Lastly and most important is a broad The Role of Veterinarians in Feral
and consistent public education program that encourages Cat Management
responsible pet ownership and promotes (1) the reasons
for keeping cats indoors or supervised while outdoors and Public policy decisions in the United States continue to be
(2) how to practically achieve this. For example, cat owners made based on inadequate information18 and influenced
need to be shown examples of devices that may be easily by loud and passionate advocacy groups. General public
used to allow cats to enjoy the outdoors without harming attitudes toward cats, experiences with feral cats, and prefer-
wildlife (e.g., cat runs/aviaries/habitats, flexible cat tunnels, ence for management should be examined across a broader
harnesses). In the absence of reproductive feral cats, new cats scale. The sociopolitical aspects of feral cat management are
enter stray populations as “lost pets” or from irresponsible the greatest challenge because the highly charged emotions
owners who abandon them. Strong enforcement of fines for associated with both sides of the issue inhibit progress on
this behavior and other measures are needed. TNR colonies actual population reduction. Additional study of public
should be relocated from and should not be established in perceptions of feral cats may help local managers make
sensitive habitats where threatened and endangered animal more informed decisions and aid in understanding the
species reside, for example, piping plover (Charadrius growing public debate regarding feral cat management.
melodus) shore habitat in Jones Beach, New York.62 Veterinarians who care about wildlife and conservation
One of the intermediate solutions that may replace free- can and should contribute to resolving the dilemma of
roaming TNR colonies is an enclosed colony. An enclosed feral cat management. First and foremost, do no harm:
colony (sometimes termed a “sanctuary”) assumes that all of when asked to participate in a TNR program, consider that
the benefits afforded a managed TNR colony are available TNR is inconsistent with the policies advocated by wildlife
for a group of cats that live in an enclosed, typically outdoor conservationists and managers. For example, the Wildlife
space.63 An enclosed colony would require a colony manager Society, a 10,000-member strong professional organization
(and the various volunteers that are typically involved in that provides board certification for wildlife biologists, and
the care of TNR colonies) to provide daily care for the is considered parallel to the American Veterinary Medical
cats, including feeding, medical care, etc. Enclosed colonies Association, strongly opposes TNR.66 Cats re-released to
would protect the welfare of cats, possibly helping them live the environment, particularly when offered supplemental
108 SE C T I O N 3 Conservation Medicine
food and shelter, continue to negatively impact wildlife. trap-neuter-return for management of free-roaming cats, J Am
Numerous other allied organizations also have strong policy Vet Med Assoc 225(12):1871–1876, 2004.
statements against TNR and in support of alternative solu- 10. Natoli E, Maragliano L, Cariola G, et al: Management of feral
domestic cats in the urban environment of Rome (Italy), Prev Vet
tions, including: the American Ornithologists’ Union, the
Med 77(3):180–185, 2006.
International Wildlife Rehabilitation Council, the Associa- 11. Castillo D, Clarke A: Trap/neuter/release methods ineffective in
tion of Avian Veterinarians, the American Association of controlling domestic cat” colonies” on public lands, Nat Areas J
Wildlife Veterinarians, the National Association of State 23(3):247–253, 2003.
Public Health Veterinarians, National Wildlife Rehabilita- 12. Baker PJ, Molony SE, Stone E, et al: Cats about town: is preda-
tors Association, and many others.67 Consider disseminat- tion by free-ranging pet cats Felis catus likely to affect urban bird
ing this message to practitioners who are often recruited populations?, Ibis 150(s1):86–99, 2008.
or hired to participate in TNR under the false assumption 13. Liberg O, Sandell M, Pontier D, et al: Density, spatial organisa-
that “doing something is better than nothing at all” and tion and reproductive tactics in the domestic cat and other felids.
explain the intricacies of why this is not true across the In Turner DC, Bateson P, editors: The domestic cat: the biology
board. Become involved in local policy; veterinarians are of its behaviour, ed 2, Cambridge, United Kingdom, 2000,
Cambridge University Press, pp 119–147.
well-respected members of the community who can have
14. Levy JK, Crawford PC: Humane strategies for controlling feral
a powerful impact at a town council meeting to determine cat populations, J Am Vet Med Assoc 225(9):1354–1360, 2004.
feral cat management strategies. Become active in educating 15. Levy JK, Gale DW, Gale LA: Evaluation of the effect of a long-
the public; engage in presentations and panels that involve term trap-neuter-return and adoption program on a free-roaming
the general public. Provide cat owners with viable solutions; cat population, J Am Vet Med Assoc 222(1):42–46, 2003.
all of us have friends who allow their cats to roam outdoors. 16. Jessup DA: The welfare of feral cats and wildlife, J Am Vet Med
Rather than cringing, provide them with designs of afford- Assoc 225(9):1377–1383, 2004.
able cat runs, inexpensive cat mesh tunnels, training tools 17. Barrows PL: Professional, ethical, and legal dilemmas of trap-
for keeping cats on a leash and other means of allowing neuter-release, J Am Vet Med Assoc 225(9):1365–1369, 2004.
supervised time outdoors as an alternative to keeping cats 18. Longcore T, Rich C, Sullivan LM: Critical assessment of claims
only indoors. Engage with others that do not share your regarding management of feral cats by trap–neuter–return,
Conserv Biol 23(4):887–894, 2009.
views and hold constructive discussions. Remember, all
19. Robertson SA: A review of feral cat control, J Feline Med Surg
stakeholders agree that the ultimate goal would be to have 10(4):366–375, 2008.
all cats under the care of a person who cares for them and 20. Lohr CA, Cox LJ, Lepczyk CA: Costs and benefits of trap-
away from harming wildlife. neuter-release and euthanasia for removal of urban cats in Oahu,
Hawaii, Cons Biol 27:64–73, 2013.
References 21. Loyd KAT, DeVore JL: An evaluation of feral cat management
options using a decision analysis network, Ecology Soc 15(4):10,
1. Lipinski MJ, Froenicke L, Baysac KC, et al: The ascent of cat 2010.
breeds: genetic evaluations of breeds and worldwide random- 22. Schmidt PM, Swannack TM, Lopez RR, et al: Evaluation of
bred populations, Genomics 91(1):12–21, 2008. euthanasia and trap–neuter–return (TNR) programs in manag-
2. Hu Y, Hu S, Wang W, et al: Earliest evidence for commen- ing free-roaming cat populations, Wildl Res 36(2):117–125,
sal processes of cat domestication, Proc Natl Acad Sci U S A 2009.
111(1):116–120, 2014. 23. Alley Cat Allies: Biology and Behavior of the Cat, 2017. https://
3. Serpell JA: Domestication and history of the cat. In Turner DC, www.alleycat.org/resources/biology-and-behavior-of-the-cat/.
Bateson P, editors: The domestic cat: The biology of its behaviour, (Accessed 28 March 2017).
ed 2, Cambridge, United Kingdom, 2000, Cambridge University 24. Williams T: Feline fatales, Audubon Magazine. 2009.
Press, pp 179–192. 25. Lauber BT, Knuth BA, Tantillo JA, et al: The role of ethical
4. Serpell J: Beneficial effects of pet ownership on some aspects of judgments related to wildlife fertility control, Soc Nat Resour
human health and behaviour, J Royal Soc of Med 84(12):717–720, 20(2):119–133, 2007.
1991. 26. PetSmart Charities: Trap-Neuter-Return Grants, https://www
5. Herzog H: The impact of pets on human health and psychologi- .petsmartcharities.org/pro/grants/spayneuter-grants/. (Accessed
cal well-being: fact, fiction, or hypothesis?, Current Dir in Psych 28 March 2017).
Sci 20(4):236–239, 2011. 27. Kays RW, DeWan AA: Ecological impact of inside/outside
6. Crooks KR, Soulé ME: Mesopredator release and avifaunal house cats around a suburban nature preserve, Anim Conserv
extinctions in a fragmented system, Nature 400(6744):563–566, 7(3):273–283, 2004.
1999. 28. Lepczyk CA, Mertig AG, Liu J: Landowners and cat predation
7. Loyd KAT, Hernandez SM: Public perceptions of domestic cats across rural-to-urban landscapes, Biol Conserv 115(2):191–201,
and preferences for feral cat management in the southeastern 2004.
United States, Anthrozoos 25(3):337–351, 2012. 29. Nogales M, Martín A, Tershy BR, et al: A review of feral cat
8. Marra PP, Santella C: Cat wars: the devastating consequences of a eradication on islands, Conserv Biol 18(2):310–319, 2004.
cuddly killer, 2016, Princeton University Press. 30. Adamec RE: The interaction of hunger and preying in the
9. Andersen MC, Martin BJ, Roemer GW: Use of matrix domestic cat (Felis catus): an adaptive hierarchy?, Behav Biol
population models to estimate the efficacy of euthanasia versus 18(2):263–272, 1976.
CHAPTER 18 Feral Cat Dilemma 109
31. Barratt D: Predation by house cats, Felis catus (L.), in Canberra, 50. McElroy KM, Blagburn BL, Breitschwerdt EB, et al: Flea-
Australia. II. Factors affecting the amount of prey caught and associated zoonotic diseases of cats in the USA: bartonellosis,
estimates of the impact on wildlife, Wildl Res 25(5):475–487, flea-borne rickettsioses, and plague, Trends Parasitol 26(4):197–
1998. 204, 2010.
32. Davis DE: The use of food as a buffer in a predator-prey system, 51. Torrey EF, Yolken RH: Schizophrenia and toxoplasmosis,
J Mammal 38(4):466–472, 1957. Schizophr Bull 33(3):727–728, 2007.
33. Doherty TS, Glen AS, Nimmo DG, et al: Invasive predators 52. Dalimi A, Abdoli A: Latent toxoplasmosis and human, Iran J
and global biodiversity loss, Proc Natl Acad Sci USA 113(40): Parasitol 7(1):1, 2012.
11261–11265, 2016. 53. Childs JE: Urban cats: their demography, population density,
34. Loss SR, Will T, Marra PP: The impact of free-ranging domestic and owner characteristics in Baltimore, Maryland, Anthrozoos
cats on wildlife of the United States, Nat Commun 4:1396, 3(4):234–244, 1990.
2013. 54. Cole D, Atkins L: Rabies: the epidemic continues in Georgia,
35. Spotte S: Free-ranging cats: behavior, ecology, management, 2014, Ga Epidemiol Rep 23:1–4, 2007.
John Wiley & Sons. 55. Roseveare CW, Goolsby WD, Foppa IM: Potential and actual
36. Grimm NB, Faeth SH, Golubiewski NE, et al: Global change terrestrial rabies exposures in people and domestic animals,
and the ecology of cities, Science 319(5864):756–760, 2008. upstate South Carolina, 1994–2004: A surveillance study, BMC
37. Nutter FB, Levine JF, Stoskopf MK: Reproductive capacity of Public Health 1:65, 2009.
free-roaming domestic cats and kitten survival rate, J Am Vet Med 56. Gerhold R, Jessup D: Zoonotic diseases associated with free-
Assoc 225(9):1399–1402, 2004. roaming cats, Zoonoses Public Health 60(3):189–195, 2013.
38. Norris JM, Bell ET, Hales L, et al: Prevalence of feline immuno- 57. Lepczyk CA, Lohr CA, Duffy DC: A review of cat behavior in
deficiency virus infection in domesticated and feral cats in eastern relation to disease risk and management options, Appl Anim
Australia, J Feline Med Surg 9(4):300–308, 2007. Behav Sci 173:29–39, 2015.
39. Nassar R, Mosier JE: Feline population dynamics: a study 58. Carver S, Bevins SN, Lappin MR, et al: Pathogen exposure varies
of the Manhattan, Kansas, feline population, Am J Vet Res widely among sympatric populations of wild and domestic felids
43(1):167–170, 1982. across the United States, Ecol Appl 26(2):367–381, 2016.
40. Warner RE: Demography and movements of free-ranging 59. Cunningham MW, Brown MA, Shindle DB, et al: Epizootiology
domestic cats in rural Illinois, J Wildl Manage 340–346, 1985. and management of feline leukemia virus in the Florida puma, J
41. Loyd KAT, Hernandez SM, Abernathy KJ, et al: Risk behaviours Wildl Dis 44(3):537–552, 2008.
exhibited by free-roaming cats in a suburban US town, Vet Rec 60. Cole R, Lindsay D, Howe D, et al: Biological and molecular char-
173(12):295, 2013. acterizations of Toxoplasma gondii strains obtained from southern
42. People for the Ethical Treatment of Animals: Feral cats: Trap- sea otters (Enhydra lutris nereis), J Parasitol 86(3):526–530, 2000.
ping is the Kindest Solution, 2017. http://www.peta.org/issues/ 61. Dubey JP: Toxoplasmosis of animals and humans, 2016, CRC
companion-animal-issues/companion-animals-factsheets/feral- Press.
cats-trapping-kindest-solution/. (Accessed 28 March 2017). 62. Fry C: Piping Plover Defenders Gear Up for Trial, The Courthouse
43. National Wildlife Rehabilitors Association: “Facts about NWRA” News Service, Feb 7 2017. https://www.courthousenews.com/
2017. http://www.nwrawildlife.org/?page=Facts. (Accessed 28 piping-plover-defenders-gear-up-for-trial/. (Accessed 28 March
March 2017). 2017).
44. Schenk AN, Souza MJ: Major anthropogenic causes for and 63. Kortis B: Implementing a community Trap-Neuter-Return
outcomes of wild animal presentation to a wildlife clinic in East Program, The Humane Society of the United States. http://www
Tennessee, USA, 2000–2011, PLoS ONE 9(3):e93517, 2014. .humanesociety.org/assets/pdfs/pets/implementing_community
45. Ancillotto L, Serangeli MT, Russo D: Curiosity killed the bat: _tnr_pt1.pdf. (Accessed 28 March 2017).
domestic cats as bat predators, Mamm Biol 78(5):369–373, 64. Centonze LA, Levy JK: Characteristics of free-roaming cats and
2013. their caretakers, J Am Vet Med Assoc 220(11):1627–1633, 2002.
46. Koenig J, Shine R, Shea G: The dangers of life in the city: 65. The Australian Government: Department of the Environment
patterns of activity, injury and mortality in suburban lizards and Energy, Tackling Feral Cats. 2015. http://www.environment
(Tiliqua scincoides), J Herpetol 36(1):62–68, 2002. .gov.au/biodiversity/threatened/publications/factsheet-tackling
47. Mcruer DL, Gray LC, Horne L, et al: Free-roaming cat interac- -feral-cats. (Accessed 28 March 2017).
tions with wildlife admitted to a wildlife hospital, J Wildl Manage 66. The Wildlife Society: Issue Statement, Feral and Free-roaming
81(1):163–173, 2016. Domestic Cats. 2016. http://wildlife.org/wp-content/uploads/
48. Loyd KAT, Hernandez SM, Mcruer DL: The role of domestic 2014/05/PS_FeralandFreeRangingCats.pdf. (Accessed 28 March
cats in the admission of injured wildlife at rehabilitation and 2017).
rescue centers, Wildl Soc Bull 2017. (online only at print). 67. The American Bird Conservancy: Trap Neuter Release, https://
49. Dabritz H, Conrad PA: Cats and Toxoplasma: implications for abcbirds.org/program/cats-indoors/trap-neuter-release/.
public health, Zoonoses Public Health 57(1):34–52, 2010. (Accessed 19 April 2017).
19
The United States Agency for
International Development Emerging
Pandemic Threats PREDICT Project—
Global Detection of Emerging Wildlife
Viral Zoonoses
KIRSTEN V.K. GILARDI AND JONNA A.K. MAZET
110
CHAPTER 19 The United States Agency for International Development Emerging Pandemic Threats PREDICT Project 111
• Figure 19.1 Wildlife Viral Pathogen Spillover Transmission of emerging zoonotic pathogens from
wildlife populations (pink) into livestock (green) and/or people (red) may lead to devastating outbreaks.
(Modified from Karesh WB, Dobson A, Lloyd-Smith JO, et al: The ecology of zoonoses: natural and
unnatural histories. Lancet 380:1936–1943, 2012.)
live in markets for human consumption. This unfortunately from wildlife sources that would inform capacity-building
triggered large-scale extermination of civets in a misguided for EID mitigation, control, and prevention. Led by the
attempt to curb the outbreak, as it was later determined that University of California, Davis, One Health Institute (in
SARS-like coronaviruses (Co-V) were endemic in wild bats, the School of Veterinary Medicine), PREDICT was initially
with civets and humans as spillover hosts.14 implemented in 20 countries in Africa, South and Southeast
Asia, and Latin America by a consortium of organizations
including EcoHealth Alliance, Metabiota, the Smithsonian
United States Agency for International Institution, and the Wildlife Conservation Society (Fig.
Development Emerging Pandemic Threats 19.2). PREDICT’s objectives were to: (1) conduct wildlife
PREDICT Project (2009–2014) surveillance and virus discovery in EID “hotspots” char-
acterized by high wildlife diversity and increasing human
The US Agency for International Development (USAID) pressure on natural resources; (2) characterize high-risk
has been working globally to improve both our collective human-animal interfaces, behaviors, and drivers of pathogen
understanding of the risk for transmission of pathogens spillover from wildlife to people; (3) improve virus detec-
from wildlife to humans, and to increase the capacity in tion and discovery by developing laboratory and disease
developing countries for effective response and contain- outbreak response capacities; (4) optimize predictive models
ment. Expanding upon its substantial investments in build- for zoonotic disease emergence and spread; and (5) deploy
ing worldwide capacity for avian influenza surveillance and cutting-edge information management and communication
control, USAID launched the Emerging Pandemic Threats tools to advance a more integrated, global approach to
(EPT) program in 2009 with the goal of strengthening sharing data from zoonotic virus surveillance. As well, the
capacities in developing countries to prevent, detect, and PREDICT Consortium was wholly committed to limiting
control all emergent wildlife zoonotic diseases (not just potential harm to wildlife populations by implementing
avian influenza). The EPT program was designed to be pro- live-capture protocols only and by expressing a conserva-
active and preemptive in its approach to wildlife zoonoses: tion ethic in all activities, especially communications with
to increase our understanding of the ecologic, viral, and stakeholders. For example, when discussing bats as hosts
behavioral drivers of wildlife virus spillover; train people of potential zoonoses, PREDICT teams always coupled
in biodiversity hotspots around the world to detect and these messages with an explanation of the importance of
control zoonoses and disease emergence; and build capacity bats to ecosystems, agriculture, and biodiversity. Also, in
among government ministries for outbreak response and order to discourage bat depopulation, messages emphasized
mitigation.15 that killing bats would likely result in increased risk for
Of several core projects that comprised the first 5-year viral transmission due to dispersal of disturbed popula-
phase of the EPT (2009–2014), PREDICT’s mandate was tions and increases in reproductive rates to compensate for
to build the evidence base for zoonotic disease emergence mortality.
112 SE C T I O N 3 Conservation Medicine
• Figure 19.2 PREDICT Consortium and Countries from 2009 to 2014, a consortium comprised of the
University of California, Davis, EcoHealth Alliance, Metabiota, Wildlife Conservation Society, and Smithson-
ian Institution implemented the USAID Emerging Pandemic Threats PREDICT project in 20 countries in
Africa, South and Southeast Asia, and Latin America. (From PREDICT Consortium: Reducing pandemic
risk, promoting global health. One Health Institute, University of California, Davis, December 2014. http://
www.vetmed.ucdavis.edu/ohi/local_resources/pdfs/chapters/6_predict_virus_detection_discovery.pdf)
wildlife samples. This approach allowed for the detection of Novel Viruses (Table 19.1)
viruses presumably present at low levels in the populations
surveyed, while also enabling a broad search for known and Among hundreds of viral discoveries, many have implica-
new viruses in the range of zoonotic viral families, includ- tions as potential sources of pandemics of wildlife origin. For
ing alphaviruses, arenaviruses, astroviruses, bunyaviruses example, PREDICT effectively doubled the known number
(including hantaviruses), coronaviruses, filoviruses, flavi- of viruses in the family Coronaviridae that includes SARS
virues, herpesviruses, orthomyxoviruses, paramyxoviruses, and Middle Eastern Respiratory Syndrome (MERS) (see
poxviruses, reoviruses, retroviruses, and rhabdoviruses. The Chapter 42). Novel retroviruses and paramyxoviruses were
use of cPCR was appropriate technology for use by PRE- also detected, including the detection of novel henipaviruses
DICT laboratories around the world, as it was relative- (the same viral family that contains Nipah and Hendra
ly inexpensive and easy to implement in resource-limited viruses) and filovirus exposure in bats.19–24 As well, PREDICT
countries. Samples identified by PCR were further cloned researchers found strong evidence for western lowland goril-
and sequenced, enabling discernment between previously las (Gorilla gorilla gorilla) as the nonhuman primate reservoir
described and new viruses. for Human T-lymphotropic virus in western Africa, and
discovered a new simian immunodeficiency virus strain in
Information Management and Reporting a naturally infected chimpanzee (Pan troglodytes troglodytes)
with AIDS-like symptoms.25,26 Hepatitis B virus (HBV) was
In order to handle the enormous quantity of wildlife sur- found to be circulating among gorillas and chimpanzees and
veillance, site characterization, and laboratory testing data among subspecies of chimpanzees, refuting the previously
generated by all 20 countries, PREDICT developed an held assumption that HBV genotypes are host-specific,
in-house on-line platform for secure, internal, standardized with implications for the potential for spillover in this
and centralized data collection, collation, and management. group of viruses.27 Furthermore, PREDICT documented
Policies on data ownership, governance, and release were anthropozoonoses—human to primate transmission of
developed in collaboration with host governments and viruses—including a human metapneumovirus that caused
tailored to each country. Once laboratory test results were fatal respiratory disease in wild mountain gorillas (Gorilla
verified and interpreted by the global viral discovery team, beringei beringei) and human herpes simplex-1 virus that
test results reports were shared exclusively with host country caused classic stomatitis in confiscated eastern lowland
governments, and any findings of potential concern were gorillas (Gorilla beringei graueri).28,29
discussed with governments first. Once host governments
had an adequate period of time to receive and consider the New Models for Emergence
potential implications of PREDICT findings, data were
made publically accessible through the public data-sharing PREDICT built upon the original “hotspots” model to
and visualization platform, HealthMap.18 further assess patterns of wildlife disease emergence in time
and space, confirming that the risk of EIDs is highest in
PREDICT Results areas of high mammalian biodiversity, and finding that
mammal diversity, land use type, and land use change are
PREDICT is likely the most comprehensive wildlife viral the most important factors for predicting wildlife EIDs.3,30
detection and zoonotic disease capacity development As well, PREDICT searched all published data available
program in the world to date. It achieved major advances in through 2010 to identify animal hosts, human activities,
understanding wildlife viruses and the factors that contribute and high-risk disease transmission interfaces implicated
to their spillover into human populations on a global scale in zoonotic virus spillover. Network analyses were used to
and in building capacity in less developed countries for the examine these data for transmission pathways, viral traits,
rapid detection and control of EIDs (Fig. 19.3). Through host species characteristics, taxonomic ranges, and geo-
PREDICT, more than 2500 people, including government graphic distributions of zoonotic viruses, to provide fresh
personnel, veterinarians, students, physicians, laboratory insight on the virus characteristics and conditions that pose
technicians, field biologists, and hunters, were trained a risk for future wildlife EIDs.16
in biosafety and PPE, surveillance methods, laboratory
techniques, and disease outbreak investigation. PREDICT Outbreak Response
worked with more than 32 diagnostic laboratories around
the world to institute low-cost methodologies for conduct- In addition to conducting wildlife viral discovery,
ing PCR assays for rapid detection of viruses in wildlife PREDICT provided support to governments and institu-
samples. By humanely sampling more than 56,000 nonhu- tions during 23 zoonotic disease outbreaks in 10 countries
man primates, bats, rodents, and other wildlife at high-risk between 2010 and 2014, most of which were impacting
human-wildlife interfaces, PREDICT detected 984 unique human populations. For example, PREDICT collected bat
viruses in wild animals and people, 815 of which were samples and integrated wildlife and human response teams
novel. This effort more than doubled the number of known to help respond to a Nipah virus outbreak in Bangladesh
mammalian viruses in the world. and participated in several Ebolavirus outbreak response
114 SE C T I O N 3 Conservation Medicine
• Figure 19.3 PREDICT Results The first 5-year phase of the USAID Emerging Pandemic Threats
PREDICT was the most comprehensive zoonotic disease surveillance and capacity-building effort ever
undertaken; it trained 2500 people, built capacity in 32 laboratories, humanely sampled more than
56,000 wild animals, and detected 984 viruses, the majority of which were new discoveries. (From
PREDICT Consortium: Reducing pandemic risk, promoting global health. One Health Institute, University
of California, Davis, December 2014. http://www.vetmed.ucdavis.edu/ohi/local_resources/pdfs/chapters/
6_predict_virus_detection_discovery.pdf)
efforts in Uganda by collecting samples from wildlife and communities. During the devastating 2014–2015 West
domestic livestock and conducting surveys in Ebola-affected Africa Ebolavirus outbreak, PREDICT achieved the early
communities to better understand how people were coming detection of an Ebola virus causing an outbreak in the
into contact with wildlife. In 2012, PREDICT provided Equateur Province, Democratic Republic of Congo, which
post-mortem and field investigation support to diagnose arose independently of the West Africa outbreak. Rapid
a yellow fever outbreak causing mortality in red howler and accurate detection of the virus facilitated early control
monkeys in Peru, in time for the government to mount procedures by the government, which effectively contained
a vaccination program to prevent spread to local human the outbreak before it could spread.
CHAPTER 19 The United States Agency for International Development Emerging Pandemic Threats PREDICT Project 115
TABLE
19.1 PREDICT Viral Discovery
Note numbers of viruses do not total to 984 as viruses have been found in more than one wildlife host taxa.
From PREDICT Consortium: Reducing pandemic risk, promoting global health. One Health Institute, University of California, Davis, December 2014. http://www.
vetmed.ucdavis.edu/ohi/local_resources/pdfs/chapters/6_predict_virus_detection_discovery.pdf.
Next Steps: PREDICT 2014–2019 to government partners and interactions with communities
about PREDICT created unique opportunities to allay fears
The first phase of PREDICT realized significant advances about or animosity toward wildlife and to talk about the
in our knowledge of the global wildlife virome and the intrinsic value of wildlife populations, intact habitats, and
human activities and land use changes that put people at biodiversity.
greater risk for spillover infections from wildlife viruses.31 In its current phase (2014–2019), PREDICT has
PREDICT served as a real-world proof of concept of the embarked upon an even larger scope with a more intense
relevance and appropriateness of addressing disease risk at focus on the dynamics of zoonotic viruses in wildlife, people,
the human-animal-environment or One Health interface and livestock (primarily influenza, filovirus, paramyxovirus,
and achieved it with a conservation ethos.32 Presentations and coronavirus) and the human behaviors that drive their
116 SE C T I O N 3 Conservation Medicine
spillover, amplification, and spread. Working with new gov- 14. Li W, Shi Z, Yu M, et al: Bats are natural reservoirs of SARS-like
ernmental and nongovernmental organization partners in Coronaviruses, Science 310(5748):676–679, 2005.
30 countries, the team is documenting viral sharing among 15. United States Agency for International Development: Emerg-
ing Pandemic Threats Program, 2013; http://www.usaid.gov/
diverse hosts and targeting even more intense surveillance
news-information/fact-sheets/emerging-pandemic-threats
at high-risk pathways for viral transmission to identify the -program.
social and ecological drivers of pathogen emergence. In 16. Johnson CK, Hitchens PL, Smiley Evans T, et al: Spillover and
particular, PREDICT is addressing the risk of wildlife sold pandemic properties of zoonotic viruses with high host plasticity,
as food and medicine in markets in Asia and Africa, explor- Sci Rep 5:14830, 2015, doi:10.1038/srep14830.
ing both the magnitude of the conservation and disease 17. Evans TS, Gilardi KVK, Barry P, et al: Detection of viruses
risks, as well as the palatability of specific interventions to using discarded plants from wild mountain gorillas and golden
motivate behavior change. Ultimately, the goal is to provide monkeys, Am J Primatol 2016, doi:10.1002/ajp.22576.
essential information for helping less developed nations 18. PREDICT HealthMap: www.healthmap.org/PREDICT.
strengthen their capacities for epidemic prevention, thereby 19. Anthony SJ, Ojeda-Flores R, Rico-Chávez O, et al: Coronavi-
contributing to pandemic prevention and improving global ruses in bats in Mexico, J Gen Virol 94:1028–1038, 2013.
20. Anthony SJ, Gilardi K, Goldstein T, et al: Further evidence for
health security.
bats as the evolutionary source of MERS Coronavirus. MBio.
Submitted.
References 21. Wacharapluesadee S, Sintunawa C, Kaewport T, et al: Group C
Betacoronavirus in bat guano fertilizer, Thailand, Emerg Infect
1. Fauci AS: Emerging and re-emerging infectious diseases: the Dis 19(8):1349–1351, 2013.
perpetual challenge, Acad Med 80(12):1079–1085, 2005. 22. Yuan L, Li M, Li L, et al: Evidence for retrovirus and para-
2. Daszak P, Cunningham AA, Hyatt AD: Emerging infectious myxovirus infection of multiple bat species in China, Viruses
diseases of wildlife – threats to biodiversity and human health, 6(5):2138–2154, 2014.
Science 287:443–449, 2000. 23. Weiss S, Nowak K, Fair J, et al: Henipavirus-related sequences
3. Jones KE, Patel NG, Levy MA, et al: Global trends in emerging in fruit bat bushmeat, Republic of Congo, Emerg Infect Dis
infectious diseases, Nature 451:990–994, 2008. 18(9):1536–1537, 2012.
4. Morse SS: Factors in the emergence of infectious diseases, Emerg 24. Olival KJ, Islam A, Yu M, et al: Ebola virus antibodies in fruit
Infect Dis 1:7–15, 1995. bats, Bangladesh, Emerg Infect Dis 19(2):270–273, 2013.
5. Calisher CH, Childs JE, Field HE, et al: Bats: important reser- 25. LeBreton M, Switzer WM, Djoko CF, et al: A gorilla reservoir for
voir hosts of emerging viruses, Clin Microbiol Rev 19:531–545, human T-lymphotropic virus type 4 (HTLV-4), Emerg Microbes
2006. Infect 3(1):e7, 2014, doi:10.1038/emi.2014.7.
6. Gao F, Bailes E, Robertson DL, et al: Origin of HIV-1 in the 26. Etienne L, Nerrienet E, LeBreton M, et al: Characterization
chimpanzee Pan troglodytes troglodytes, Nature 397:436–441, of a new simian immunodeficiency virus strain in a naturally
1999. infected Pan troglodytes troglodytes chimpanzee with AIDS related
7. Almeida MAB, Cardoso JC, dos Santos E, et al: Surveillance for symptoms, Retrovirology 8(4):4, 2011.
Yellow Fever Virus in non-human primates in Southern Brazil, 27. Lyons S, Sharp C, LeBreton M, et al: Species association of
2001–2011: a tool for prioritizing human populations for vac- hepatitis B virus (HBV) in nonhuman apes: evidence for recom-
cination, PLoS Negl Trop Dis 8(3):e2741, 2014, doi:10.1371/ bination between gorilla and chimpanzee variants, PLoS ONE
journal.pntd.0002741. 7(3):e33430, 2012.
8. Schmaljohn C, Hjelle B: Hantaviruses: a global disease problem, 28. Palacios G, Lowenstine LJ, Cranfield MR, et al: Human
Emerg Infect Dis 3(2):95–104, 1997. metapneumovirus infection in wild mountain gorillas, Rwanda,
9. Hotez PJ, Dumonteil E, Woc-Colburn L, et al: Chagas disease: Emerg Infect Dis 17(4):711–713, 2011.
the new HIV/AIDS of the Americas, PLoS Negl Trop Dis 29. Gilardi KVK, Oxford KL, Gardner-Roberts D, et al: Human
6(5):e1498, 2012, doi:10.1371/journal.pntd.0001498. herpes simplex virus type 1 in confiscated gorilla, Emerg Infect
10. Keesing F, Belden LK, Daszak P, et al: Impacts of biodiversity Dis 20(11):1883–1886, 2014.
on the emergence and transmission of infectious diseases, Nature 30. PREDICT Consortium: Modeling & analytics. In Reducing pan-
468:647–652, 2010. demic risk, promoting global health, Davis, December 2014, One
11. Karesh WB, Dobson A, Lloyd-Smith JO, et al: The ecology of Health Institute, University of California, pp 96–117. Available
zoonoses: natural and unnatural histories, Lancet 380:1936–1943, at: http://www.vetmed.ucdavis.edu/ohi/predict/final_report.
2012. 31. PREDICT Consortium: Reducing pandemic risk, promoting global
12. Chua KB, Goh KJ, Wong KT, et al: Fatal encephalitis health, Davis, December 2014, One Health Institute, University
due to Nipah virus among pig-farmers in Malaysia, Lancet of California. Available at: http://www.vetmed.ucdavis.edu/ohi/
354(9186):1257–1259, 1999. predict/final_report.
13. Zhong NS, Zheng BJ, Li YM, et al: Epidemiology and cause 32. Kelly TR, Karesh WB, Johnson CK, et al: One Health proof of
of severe acute respiratory syndrome (SARS) in Guangdong, concept: bringing a transdisciplinary approach to surveillance
People’s Republic of China, in February 2003, Lancet 462(9393): for zoonotic viruses at the human-wild animal interface, Prev Vet
1353–1358, 2003. Med 137:112–118, 2017.
20
Renewable Energy: Effects on Wildlife
PEREGRINE L. WOLFF AND MICHAEL R. SCHIRMACHER
117
118 SE C T I O N 3 Conservation Medicine
When developing impact minimization measures and wind facilities that are designed and operated with utmost
the required permits necessary for a new project, agency care to conserve wildlife may, under some circumstances,
personnel consider project type, project size, land owner- result in the “unavoidable” take of eagles. Wind project
ship, proximity to other developments, and the presence operators must implement eagle conservation practices
of protected or sensitive state or federal fish and wildlife that are required for the issuance of Eagle Incidental Take
species. A case-by-case approach is used to determine the Permits.9
focus of applicable regulations and impact minimization. Project 3: Utility-scale concentrating solar power project
Three examples of renewable energy projects in Nevada are located on public land managed by BLM. Concentrated or
provided, which illustrate the application of either state or thermal solar power plants use tracking mirrors to reflect
federal regulations. and concentrate sunlight onto a heat exchanger or receiver
Project 1: Utility-scale solar PV plant that is proposed tubes. The tubes contain fluids that absorb the concentrated
on public land managed by the BLM in the Mojave Desert sunlight in the form of heat energy. The heated fluids flow
within occupied desert tortoise (Gopherus agassizii) habitat. through the tubes and are used to produce steam that drives
A potential effect on the desert tortoise due to solar develop- a conventional turbine, which is connected to a generator
ment is direct mortality from vehicles or equipment that producing electricity. Similar to coal-fired or natural gas
may collapse burrows or entrap tortoises within burrows.5 power plants, water is necessary in the cooling processes and
The desert tortoise is listed under the US Endangered is discharged to industrial wastewater evaporation ponds. In
Species Act (ESA) as threatened. Section 7 of the ESA the deserts of the southwest, these ponds are an attractant
is the mechanism by which federal agencies ensure the to migratory and nonmigratory birds, and potentially bats,
actions they take, including those they fund or authorize, and may represent a secondary hazard if the wastewater
do not jeopardize the existence of any listed species. If contains toxicants or the increase in the density of birds
through a biological assessment or other review process, increases collision mortalities with mirrors and the tower.
the BLM determines its action (authorizing the PV project) State agencies, such as the Nevada Department of Wild-
is “likely to adversely affect” the desert tortoise, the BLM life (NDOW), may regulate these artificial or artificially
then submits to the USFWS a request for formal consulta- created bodies of water that contain chemicals or substances
tion. During this consultation, the USFWS will prepare that cause or will cause the death of wildlife. NDOW’s
a biological opinion on whether the proposed solar PV Industrial Artificial Pond permit is a means to facilitate
project will jeopardize the continued range-wide existence impact avoidance, minimization, and mitigation measures
of the desert tortoise. regarding industrial solutions that pose a risk to wildlife and
In this example, the USFWS finds that the PV project are usually in an open environment like a pond. Site-specific
may adversely affect the desert tortoise but not jeopardize recommendations for reducing impacts to wildlife are made
its continued existence. When this happens, the USFWS on a case-by-case basis. NDOW works closely with their
prepares an “incidental take statement.” Under most cir- federal partners, including the USFWS, when considering
cumstances, the ESA prohibits take, which is defined as measures proposed for species protected not only under the
harming (including killing) or harassing a listed species. MBTA, but also under the State of Nevada.10
Take that results from a federal action but is not the purpose
of the action is considered an incidental take. Incidental Wildlife Impacts From Specific
take may be allowed when the USFWS approves it through
an incidental take statement. The statement establishes Energy Sources
the amount or extent of anticipated take, reasonable and Wind Energy
prudent measures to minimize the take, and terms and
conditions that must be observed when implementing those Wind energy facilities (WEF) are either on- or offshore.
measures.6 Onshore WEF are one of the cheapest and most developed
Project 2: Utility-scale wind farm proposed on public renewable energy sources and currently account for 0.5%
land managed by the BLM in the Great Basin within close of global energy production.2 The Intergovernmental Panel
proximity to golden eagle (Aquila chrysaetos) nesting ter- on Climate Change predicts that this will increase by
ritories. The primary threat to eagles at wind facilities is 5%–29% by 2030. Wind energy has probably generated
collisions with the towers and turbine blades.7 The Migra- the most studies on impacts to wildlife, with the majority
tory Bird Treaty Act (MBTA)8 and Bald and Golden Eagle conducted in the areas where the facilities are located, in
Protection Act8 prohibit take of eagles except pursuant to North America and Europe.
federal regulations. In 2009, the USFWS promulgated new The development of offshore installations is increasing
permit rules for eagles that address the issue of wind farms.9 with a trend for turbines farther from shore in deeper
Under these new rules the USFWS can issue permits that waters.11 The primary environmental concerns identified
authorize individual instances of take of bald (Haliaeetus include: (1) the effects of increased noise levels on marine
leucocephalus) and golden eagles or “programmatic” take, mammals, turtles, and fish; (2) collision risks for seabirds
which means that instances of “take” may not be isolated with the turbines and tower structure; and marine mammals
but may recur.9 These new rules acknowledge that even and sea turtles with increased boat traffic; (3) impacts to
CHAPTER 20 Renewable Energy: Effects on Wildlife 119
benthic and pelagic habitats including food webs; (4) pollu- in bats can help to determine if this is a significant source of
tion from the release of seabed contaminants and construc- fatality. In addition, studies have suggested that bat fatalities
tion vessels; and (5) disruption for all species to migration do not seem to be random events, and available data suggest
routes and feeding grounds.11,12 The construction phase of that bats—at least some species—might be attracted to
the turbines potentially leads to the greatest impacts on wind turbines.20 In both the United States and Canada, at
wildlife from noise and increased vessel traffic. Driving piles least 22 species of bats have been reported as fatalities at
into the seafloor is loud with sounds audible under water WEF.15 However, certain species appear to be more suscep-
for tens of kilometers. There is concern that this generated tible than others. For example, three species of migratory
noise could cause damage to hearing in addition to disrupt- tree-roosting bats, hoary bats (Lasiurus cinereus), eastern red
ing communication and leading to spatial displacement in bats (Lasiurus borealis), and silver-haired bats (Lasionycteris
marine mammals and perhaps sea turtles.11 Studies have noctivagans), make up nearly 80% of the reported fatalities
shown a number of marine fish species to be affected by at sites in the United States and Canada.18 The composition
noise levels equivalent to pile driving, documenting baro- of fatalities may vary geographically, with high proportions
trauma injury and hearing loss as well as behavior changes, of Brazilian free-tailed bat (Tadarida brasiliensis) fatalities
which could, potentially, disrupt movements for breeding, reported in the southwestern United States.21 The impact of
spawning and foraging.13 Once operational, undersea noise wind turbines on other species, including the tri-colored bat
levels from the turbines are considered low enough as to (Perimyotis subflavus), Indiana bat (Myotis sodalis), northern
not cause severe noise-related issues in marine mammals. long-eared bat (Myotis septentrionalis), and little brown bat
However, it is not known whether turtles, fish, or other (Myotis lucifugus), although found in relatively low numbers
benthic species may be impacted by lower frequency noise in the United States and Canada, is a concern because
levels or by electromagnetic fields generated from cables populations are already decimated by white-nose syndrome,
that transfer the electricity to shore.11,13 a disease caused by the fungus Pseudogymnoascus destructans
The effects on avian species from offshore WEF have (see Chapter 72).22 Because bats have low reproductive
been assessed as (1) risk of collision, (2) disturbance, or (3) rates, large-scale fatalities may result in a population-level
loss of foraging habitat, and are highly variable by species.12 impact.23 Recent research suggests that the hoary bat may
The risk of collision correlates with species who do not suffer 90% population decline in the next 50 years if fatali-
avoid wind farms and whose flight pattern is within the ties continue at the current rate,24 although this does not
sweep area of the blades, with species that fly lower to the consider large-scale implementation of impact reduction
water (below the sweep) having a decreased risk of colli- strategies, such as operational minimization (see discussion
sions.12 Disturbance results when birds spend extra time and section for more details).
energy avoiding WEF, particularly during times of increased Little is known about the impact of offshore WEF on
human activity (construction and maintenance), which may bats; however, migrating bats have been documented off-
lead to displacement, potentially depriving them of impor- shore and further investigation is warranted to understand
tant foraging areas.12 Decreased use by some species of areas how offshore developments may affect bats.11
within 100–600 m from WEF has been documented in a
number of studies.14 Discussion
Bat fatalities at wind turbines were first documented The majority of studies on renewable energy’s impact on
from Australia15 in the 1970s, but received little attention wildlife results from studies on avian mortalities at onshore
until nearly three decades later when estimates of thousands WEF. Similar to offshore WEF, there are concerns for
of bat fatalities from individual WEF were reported in the impacts during both the construction and the operation
eastern United States.16 At that time, the installed wind phases of these facilities, with multispecies studies docu-
energy capacity was approximately 6 gigawatts (GW); now, menting highly variable responses of species to both phases.
through the first quarter of 2017, it exceeds 84 GW.17 When compared to reference sites, in a European-based
Cumulative estimates for the United States and Canada study, there was a documented decline in the abundance of
suggest that hundreds of thousands of bats are killed by 10 diverse bird species during construction.25 Some species
wind turbines each year.18 returned to preconstruction density during the operation;
Direct collision with moving turbine blades as well as however, others remained suppressed or absent at sites.25
barotrauma due to sudden pressure changes near the moving What is not clear from many of these studies is if displaced
blades are both thought to contribute to mortality in bats. birds are moving to other available habitats for breeding, or
Gross necropsy, histopathology, and radiology have been are being lost from local breeding populations.25 There is
used to examine carcasses recovered from around turbines.19 also some evidence that the impact on local avian population
Lesions consistent with blunt force trauma (fractures, densities around WEF may decrease with time, thus short-
abdominal wall herniation, hemothorax, and moderate term monitoring studies (<5 years) and may not be captur-
to severe middle ear hemorrhage) as well as barotrauma ing the true impact on populations.26 There is currently no
(hemothorax and moderate to severe middle ear hemor- published information on avoidance or disturbance effects
rhage) have been documented,19 although more informa- of WEF on large mammals.27 Approximately 250 species
tion on the pressure changes required to cause barotrauma of birds, primarily small passerines, make up the majority
120 SE C T I O N 3 Conservation Medicine
of the direct (collision-related) mortalities from terrestrial culminated in the hypothesis that bat fatality may be reduced
WEF; however, large raptors have received the greatest by preventing blades from spinning during high-risk condi-
public attention. The Altamont Pass Wind Resource Area tions. This is accomplished by feathering the blades (i.e.,
(APWRA) in Alameda County, California, is the oldest pitching the blades parallel to the wind) until wind speeds are
of the WEF in the United States. Brought on line in the high enough to reduce the risk to bats. Given the relatively
1980s, it is still the largest in the world with approximately narrow period of time when bats are at risk (i.e., at night,
6000 turbines spread over 50,000 acres.28 Reports of annual during low-wind-speed conditions, during late-summer
golden eagle mortalities (estimated at 75–110 per year) and autumn at least at latitudes ≥35°N), changes made to
brought into public conscience the deadly impact of this turbine operations are minimized. The first US-based study
new form of “green power” on avian species.28 In addition to to test this strategy demonstrated a 44%–93% reduction
golden eagles, and other raptors and passerines, the mortal- in bat fatality at turbines feathered up to wind speeds of 5
ity rates for all avian species at APWRA are estimated at and 6.5 meters/second (m/s) compared to normal turbine
4700 per year.28 Direct collisions have been documented at operating conditions of 3.0 m/s.29 In addition, this study
all WEF—the results of which are public—and reports indi- reported the annual power loss associated with this strategy
cate a range of 0 to >30 collisions/turbine/year.14 Although was ≤1%, which means it allowed 99% of the annual power
hundreds of thousands of passerines are killed at these facili- to be produced while reducing bat fatality up to 93%.
ties annually, there is no consensus among researchers that Subsequent studies have reported similar reduction levels
this level of mortality is leading to population-level impact using this strategy, although effectiveness likely varies by
in passerine species, and this represents only a small portion species, wind-speed condition, and turbine model.31 Other
of the annual human-associated cause of avian mortality.27 promising research involves the use of ultrasonic acoustic
Domestic cats (see Chapter 18), powerlines, communica- deterrents to reduce bat activity near WEF, but more studies
tion towers, buildings, and windows cause a much greater are needed before a commercially ready device is available.31
level of mortality.27 Due to their slow maturity and long Given the estimated high annual fatality of bats at WEF,
reproductive life, there are concerns for local and potential the potential population-level impact, and the relatively
species-level decline in raptors where facilities have been rapid growth of WEF, the timely large-scale adoption of
placed along migration routes—for example, in southern impact-reduction strategies is necessary to reduce risk and
Spain, where raptors congregate before migrating across the minimize fatalities.
strait of Gibraltar to Africa.27
A number of mitigation measures have been proposed Solar Photovoltaic and Concentrating Solar
and implemented to decrease the direct and indirect impact Thermal Power
of WEF on wildlife. For proposed projects, siting to avoid
areas of high conservation importance (primary migration Solar facilities (SF) may consist of hundreds to millions of
corridors and flight paths), as well as avoiding a critical solar collectors and span thousands of acres. In the United
habitat for species of high conservation concern, must be States, the largest SF are primarily located in the Desert
considered. Increasing turbine height and spacing appear to Southwest and often sited on public land. The effects of
be the critical factors at APWRA in reducing collision-related SF on wildlife are not well documented in peer-reviewed
raptor mortalities.7 The replacement of smaller low-capacity literature with the majority of reports in unpublished or
turbines at APWRA with larger, higher capacity and more non-peer-reviewed literature, such as environmental impact
efficient models is predicted to decrease collision mortalities documents.5 Similar to other forms of renewable energy, the
by 50%.27 The new, larger turbines have fewer blade rotations greatest threat to wildlife from SF may be habitat fragmen-
per minute and the towers are smooth, as opposed to the tation and loss, as well as disruption during construction
older lattice structures, which likely provided perches ideal with the impact lasting into site operation. Site preparation
for raptors scouting prey.27 Turbine shutdown or removal (grading and leveling) dust, noise, and soil compaction, as
at critical migration routes has also been implemented. well as altered water flows, alterations in microclimates, and
In an attempt to increase visibility and prevent collisions, electromagnetic field generation have all been identified
ultraviolet paint has been used on towers and blades. To as causing a potential impact on wildlife.5 Studies on the
date, limited studies have found little effectiveness in this impact of noise during construction and potentially into
method.27 European researchers have developed assessment operation have documented noise levels produced by some
tools to apply to individual projects and individual species, vehicles as reaching 110 decibels.5 At this level, Mojave
to rank the risk of a facility to priority populations and to Desert wildlife species, such as kangaroo rats (Dipodomyss
assist companies in siting offshore WEF to minimize the spp.), desert iguanas (Dipsosaurus dorsalis), and fringe-toed
impact on these species.11 lizards (Uma spp.), have been documented to suffer hearing
Current data suggest that, in general, bats are at highest loss, and abnormal emergence patterns in aestivating spade-
risk of fatality under relatively low-wind-speed conditions.29 foot toads (Scaphioous spp.) have been triggered.5
Moreover, unlike birds, bats tend not to fly into stationary In addition to wildlife, of particular concern in this
structures, and the majority of fatalities at wind turbines fragile and arid ecosystem is the impact on vital plant
occur only when the turbine blades are spinning.30 This communities. Dust from site disturbance may disrupt the
CHAPTER 20 Renewable Energy: Effects on Wildlife 121
physiologic processes of desert plants, including photosyn- peer-reviewed literature on the impacts of utility-scale
thesis, gas exchange, and water usage, whereas commonly renewable energy on wildlife, there is a desperate need for
used dust suppressants (chloride salts) applied to roads and research. No matter where you live, there is likely a renew-
graded areas may damage plant growth at the point of able energy plant in operation or in the permitting process
application as well as in adjacent habitats from runoff.5 nearby. Veterinary expertise is likely needed to help alleviate
The alteration of water flow and soil compaction also may the consequences of localized impacts to wildlife caused by
have a lasting impact on plant communities by potentially a wind farm or solar plants, transmission lines, or the toxic/
degrading significant amounts of habitat around SF sites.5 hyper-saline solutions of industrial ponds.
Many species may experience direct mortality from the Perhaps the most important action veterinarians may
construction and operation of SF; however, the desert tor- take is to embrace and teach that energy conservation is
toise, due to its threatened status listing with the USFWS, the most effective method to diminish the impact of all
has received the most attention. If incidental take is not energy-generating facilities on wildlife.
permitted, then mitigation involves translocation of the
tortoises to an alternative habitat with subsequent monitor- Acknowledgments
ing. However, the science of translocation in this species
is young, and long-term monitoring of moved tortoises Thanks to Tracy Kipke and Brad Hardenbook whose con-
greater than 2–3 years is currently lacking. Translocation of tribution to this manuscript were invaluable.
G. agassizii depends on the individual animals being located
and removed from the site. It is likely that there are a certain
number (especially juveniles) that are missed prior to site References
preparation. Additionally, soil compaction from traffic—
even from fairly lightweight vehicles—has been shown to 1. Menegaki A: Valuation for renewable energy: a comparative
extend to the depth of shallow burrows (<33 cm). This may review, Renew Sustain Energy Rev 12:2422–2437, 2008.
cause crushing or entrapment of species that use shallow 2. Anon: REN21_GSR2016_KeyFindings_en_10.pdf. Avail-
subterranean habitats for hibernation, including locally or able at: http://www.ren21.net/wp-content/uploads/2016/10/
federally protected species, such as the flat-tailed horned REN21_GSR2016_KeyFindings_en_10.pdf. (Accessed 29 May
lizards (Phrynosoma mcallii) and the Coachella Valley fringe- 2017).
toed lizard (Uma inornata).5 3. McDonald RI, Fargione J, Kiesecker J, et al: Energy sprawl or
energy efficiency: climate policy impacts on natural habitat for
Bird collisions with structures at SF, boiler towers,
the United States of America, PLoS ONE 4:e6802, 2009.
and tracking mirrors have been reported at concentrating 4. Santangeli A, Toivonen T, Pouzols FM, et al: Global change syn-
solar power facilities.32 In addition, the concentration of a ergies and trade-offs between renewable energy and biodiversity,
number of mirrors at stand-by points on the boiler tower Glob Change Biol Bioenergy 8:941–951, 2016.
may create enough heat to burn the skin and feathers of 5. Lovich JE, Ennen JR: Wildlife conservation and solar energy
birds flying through the thermal column.32 development in the desert southwest, United States, Bioscience
61:982–992, 2011.
Conclusion 6. Anon: Endangered Species Program | Laws & Policies |
Endangered Species Act | Section 7 Interagency cooperation.
Renewable energy facilities are coming online at unprec- Available at: https://www.fws.gov/endangered/laws-policies/
edented rates worldwide. According to the Renewables section-7.html. (Accessed 30 May 2017).
7. Hunt G: Golden eagles in a perilous landscape: predicting the effects
Global Status Report 2016, “2015 was an extraordinary
of mitigation for wind turbine bladestrike mortality, Sacramento,
year for renewable energy. Renewables are now cost com- California, 2002, p 72.
petitive with fossil fuels in many markets and are established 8. Anon: U.S. Fish & Wildlife Service - Migratory Bird Program |
around the world as mainstream sources of energy. Globally, Conserving America’s Birds. Available at: https://www.fws.gov/
renewable electricity production in 2015 continued to be birds/policies-and-regulations/laws-legislations/migratory-bird-
dominated by large (e.g., megawatt-scale and up) generators treaty-act.php. (Accessed 7 June 2017).
that are owned by utilities or large investors.”2 The negative 9. Anon: WEG_final.pdf. Available at: https://www.fws.gov/
impacts of climate change are one of the key drivers in ecological-services/es-library/pdfs/WEG_final.pdf. (Accessed 30
the current global commitment to bring these utility-scale May 2017).
renewable energy facilities online. However, major impacts 10. Anon: NAC: CHAPTER 503 - HUNTING, FISHING AND
to wildlife, including loss of biodiversity, may potentially TRAPPING; MISCELLANEOUS PROTECTIVE MEASURES.
Available at: https://www.leg.state.nv.us/NAC/NAC-503.html
occur in both scenarios unless there is a multistakeholder
#NAC503Sec030. (Accessed 7 June 2017).
approach or a policy supporting research to understand and 11. Bailey H, Brookes KL, Thompson PM: Assessing environmental
quantify impacts, and to improve existing or develop new impacts of offshore wind farms: lessons learned and recommen-
strategies to minimize the negative effects of utility-scale dations for the future, Aquat Biosyst 10:8, 2014.
renewable energy on wildlife. 12. Furness RW, Wade HM, Masden EA: Assessing vulnerability
There are many opportunities for veterinarians to con- of marine bird populations to offshore wind farms, J Environ
tribute to the renewable energy issue. With few published Manage 119:56–66, 2013.
122 SE C T I O N 3 Conservation Medicine
13. Thomsen F, Mueller-Blenkle C, Gill A, et al: Effects of pile 23. Barclay R, Harder L: Life histories of bats: life in the slow lane.
driving on the behavior of cod and sole. In The effects of noise on In Bat ecology, 2003, pp 209–253.
aquatic life, New York, NY, 2012, Springer, pp 387–388. 24. Frick WF, Baerwald EF, Pollock JF, et al: Fatalities at wind
14. Kuvlesky WP, Brennan LA, Morrison ML, et al: Wind energy turbines may threaten population viability of a migratory bat,
development and wildlife conservation: challenges and opportu- Biol Conserv 209:172–177, 2017.
nities, J Wildl Manag 71:2487–2498, 2007. 25. Pearce-Higgins JW, Stephen L, Douse A, et al: Greater impacts
15. Hein C, Schirmacher M: Impact of wind energy on bats: a of wind farms on bird populations during construction than
summary of our current knowledge, Hum Wildl Interact 10:19, subsequent operation: results of a multi-site and multi-species
2016. analysis, J Appl Ecol 49:386–394, 2012.
16. Arnett EB, Brown WK, Erickson WP, et al: Patterns of bat fatali- 26. Anon: Stewart-2005.pdf. Available at: https://tethys.pnnl.gov/
ties at wind energy facilities in North America, J Wildl Manag sites/default/files/publications/Stewart-2005.pdf. (Accessed 31
72:61–78, 2008. May 2017).
17. Anon: AWEA - American Wind Energy Association. Available 27. Anon: Summary of Wind-Wildlife Interactions | American
at: http://www.awea.org/. (Accessed 1 June 2017). Wind Wildlife Institute. Available at: https://awwi.org/resources/
18. Arnett EB, Baerwald EF: Impacts of wind energy development on summary-of-wind-wildlife-interactions-2/. (Accessed 17 May
bats: implications for conservation. In Adams RA, Pedersen SC, 2017).
editors: Bat evolution, ecology, and conservation, New York, 2013, 28. Anon: Avian Mortality. Golden Gate Audubon Soc. Available at:
Springer, pp 435–456. Available at: http://link.springer.com/ https://goldengateaudubon.org/conservation/birds-at-risk/avian
chapter/10.1007/978-1-4614-7397-8_21. (Accessed 1 June -mortality-at-altamont-pass/. (Accessed 31 May 2017).
2017). 29. Arnett EB, Huso MM, Schirmacher MR, et al: Altering turbine
19. Grodsky SM, Behr MJ, Gendler A, et al: Investigating the causes speed reduces bat mortality at wind-energy facilities, Front Ecol
of death for wind turbine-associated bat fatalities, J Mammal Environ 9:209–214, 2011.
92:917–925, 2011. 30. Cryan PM, Barclay RMR: Causes of bat fatalities at wind tur-
20. Cryan PM, Gorresen PM, Hein CD, et al: Behavior of bats at bines: hypotheses and predictions, J Mammal 90:1330–1340,
wind turbines, Proc Natl Acad Sci U S A 111:15126–15131, 2009.
2014. 31. Arnett EB, Hein CD, Schirmacher MR, et al: Evaluating the
21. Piorkowski MD, O’Connell TJ: Spatial pattern of summer bat effectiveness of an ultrasonic acoustic deterrent for reducing bat
mortality from collisions with wind turbines in mixed-grass fatalities at wind turbines, PLoS ONE 8:e65794, 2013.
prairie, Am Midl Nat 164:260–269, 2010. 32. McCrary MD, McKernan RL, Schreiber RW, et al: Avian mor-
22. Turner G, Reeder D, Coleman J: A five-year assessment of mor- tality at a solar energy power plant, J Field Ornithol 57:135–141,
tality and geographic spread of white-nose syndrome in north 1986.
american bats, with a look at the future. Update of white-nose
syndrome in bats, Bat Res News 13–27, 2011.
SECTION 4
Reproduction
21 Female Infertility in Zoo Animals, 124
123
21
Female Infertility in Zoo Animals
BRUCE CHRISTENSEN
W
hen investigating potential causes for lack of Vaginal strictures and septa have been diagnosed in some
success in a particular breeding program, first species that make copulation painful for the female.
determine what is known about that species with Females with these disorders typically have a history of
regard to reproductive physiology and reported causes of regular cycles, being attractive to and attracted by the male
subfertility, as well as what is known about the individual during estrus, allowing initial mating, but then objecting
animal’s history. A general list of questions for a clinical and not allowing the male to complete intromission, and
history is included (Box 21.1). This chapter will discuss may be reluctant to allow the male to mount in subsequent
potential causes for female infertility, and group them into attempts. Physical examination of the vaginal tract is neces-
stages of the reproduction process. sary to diagnose these conditions. The vaginal tract may be
evaluated manually, digitally, or through the use of specula
Female Infertility or endoscopic equipment. Visual evaluations must be done
carefully to avoid inadvertently passing a narrow instrument
When considering female reproductive health, it is useful to beyond strictures. Strictures or septa might be corrected via
categorize the stage of the reproductive process where the manual or surgical reduction. Post-procedural fibrosis with
problem is likely occurring. Ask the following questions: subsequent repeat stricture may be managed with physical
1. Does this female appear to cycle normally? emollients during the healing period, but this requires daily
2. Has early pregnancy been diagnosed and subsequently application for at least a week. If reduction is not possible,
lost? or postoperative stricture occurs, and reproduction is still
3. Has this female experienced abortion, dystocia, stillbirth, desired, then artificial insemination with planned cesarean
or neonatal losses? (Late-term pregnancy losses are section should be discussed as the vaginal anomalies would
beyond the scope of this chapter.) increase the risk of dystocia. The genetic component to
vaginal abnormalities will most likely be unknown in zoo
Causes of Subfertility Associated With species, so this will remain an uncertain risk of breeding
these animals.
Regular Cyclicity
Male Subfertility Parturition-Related Injury
When a pair of animals desired for reproduction fail to Dystocia related injuries may result in fibrous tissue forming
produce offspring, one of the most basic questions to answer inside the lumen of the reproductive tract, sometimes
is whether the fault lies with the male, the female, both, or completely occluding the lumen and trapping fluid in the
neither. If this male has recently produced offspring with a proximal portions of the tract, resulting in hydrometra,
different female, then the focus will be more on the female’s mucometra, or pyometra. Even if the fluid is resolved,
reproductive ability, compatibility between the current pair, subsequent damage to the uterus may be severe enough to
and methods being used currently compared to what has render the female infertile. Depending on the species, and
worked in the past. If the male’s current fertility is unknown, the nature of the fluid trapped, the female may or may not
he also must be the source of fertility investigation. Focus show systemic signs of disease. Bluntly dissecting the fibrous
on male subfertility is beyond the scope of this chapter but tissue and allowing the fluid to drain passively, or flushing
would minimally include a thorough reproductive history, the uterus with isotonic solutions, may achieve temporary
physical examination (with ultrasound), and semen analysis. relief. Recurrence is likely as the scar tissue is prone to form
again. Complete resolution of the problem may be achieved
Abnormal Genitalia only through hysterectomy.
Abnormalities in genital anatomy, whether congenital Dystocia may also result in damage to the cervix,
or acquired, may affect the ability of a female to mate, vestibulovaginal fold, or vulva, preventing these structures
maintain a pregnancy, or successfully undergo parturition. from acting as a physical barrier to ascending pathogens.
124
CHAPTER 21 Female Infertility in Zoo Animals 125
Surgical corrections are possible, but postoperative failure Leiomyomas in elephants reduce in size after treatment with
is a common complication. gonadotropin-releasing hormone vaccines.7
Fibrous adhesions may form within the abdomen fol- It is difficult to overstate the importance of the “use
lowing cesarean section, preventing the uterus from having it or lose it” hypothesis when considering what is known
normal contractions and making it prone to fluid accumula- from investigations of aging and reproduction in domestic
tion and an inability to maintain a pregnancy. Diagnosis is and zoo species, and the current breeding management
made by ultrasound or by exploratory surgery. Adhesions practices that have resulted in a skewed population in zoos
are likely to form again and hysterectomy may be the only toward aged, nulliparous individuals that now are having
treatment. difficulties reproducing.5 This has led to a more compre-
hensive, life-long view of reproductive health in captive
Age-Related Changes species’ breeding management. The Association of Zoos
Age-related pathologic changes to the female reproductive and Aquariums’ Reproductive Management Center now
tract will happen with most females if they live long enough, promotes lifetime reproductive planning for genetically
but they are more pronounced in older, nulliparous females, valuable females; this new direction is discussed in greater
or those with long inter-birth intervals. Pregnancy does have detail in Chapter 22. While recommendations will surely
a protective, regenerative effect on the female reproductive need to be adjusted for each species as we learn more,
tract1–6; females who experience the hormonal changes of it seems wise to plan on breeding valuable females early
multiple, nonpregnant cycles develop a variety of species- in their reproductive life at least once, and then spread-
specific pathologic changes. Age-related changes reported ing out subsequent pregnancies throughout the lifetime
in different species include cystic endometrial hyperplasia of the female. Concerns regarding surplus animals8 will
(CEH), pyometra, hydromucometra, adenomas, peri- need to be addressed and are discussed in Chapter 23 in
glandular fibrosis, periovarian cysts, and leiomyomas.1,3,5 this text.
126 SE C T I O N 4 Reproduction
be suspected.20 In mammals, sexual development involves follicular growth in the contralateral ovary. Granulosa cells
the establishment of a chromosomal sex in the embryo secrete anti-mullerian hormone (AMH), and assays measur-
(XX for females, XY for males), subsequent, respective dif- ing AMH have proven very sensitive and specific for the
ferentiation of the bipotential gonad into either ovarian or diagnosis of GCTs in horses.28 Suspicion of GCT is high
testicular tissue, and finally, under the influence of gonadal based on the finding of a large ovary with irregular tissue
hormones, differentiation of the tubular reproductive echogenicity on ultrasound. A positive AMH assay would
tract, accessory sex glands, and external genitalia. These be helpful, if normal ranges for the species in question
processes are complex, and errors at any stage will likely were also established. Exogenous anabolic steroids may
result in an individual that is functionally subfertile or negatively feedback on endogenous hormones and prevent
infertile. Often it may be impossible to define whether normal cycles. In zoo settings, if another animal in the
the individual is strictly male or female. Diagnosis of these enclosure is receiving hormone treatments, it should be
conditions starts with determining the karyotype of the investigated if any of the hormones are accessible to non-
individual. Anatomic and histopathologic evaluations of target animals and therefore affecting their reproductive
the reproductive tract are also necessary to determine the cycles.
nature of the specific condition. No fertility treatment exists
for these conditions. Very little is known of these types of Age-Related Changes
disorders in nonmammalian species. Sexual differentiation Advancing age may affect the regularity of cycles, as well
in birds is opposite to that of mammals, with the female as the subsequent fertility of those cycles. Typically these
being heterogametic (ZW) and the male homogametic age-related changes are seen in, or at least exacerbated in,
(ZZ). For reptiles, amphibians, and fish, it becomes even nulliparous females. Older, nulliparous white rhinoceros
more complicated as temperature, other environmental females (15–38 years old), for example, show prolonged
factors, or social status determine the current sex of the periods where luteal activity is absent or erratic.3,17 Simply
individual. having one calf during their early years prevents this occur-
rence of “flat-lining.” This evidence gives further support
Poor Nutrition to the already introduced idea of lifetime reproductive
The role of nutrition in a reproductive program is largely planning and allowing females to reproduce early in their
unknown. It is known that individuals at either extreme in lives in order to decrease the potential of subfertility
body condition tend to show decreased fertility. Obesity in later on.
zoo animals is sometimes a problem and may very well con-
tribute to subfertility. It is likely that some species require Anovulatory Follicles and Persistent
certain nutrients for successful or optimal reproduction, but Corpora Lutea
data on specific examples are lacking.21 Evidence has been While the etiology of anovulatory follicles is largely
presented that phytoestrogens may result in species-specific unknown, their occurrence in many domestic species is
reproductive losses—for example, causing subfertility well documented. Typically these follicles grow larger
in captive-born female white rhinoceros, but not female than the typical size for dominant follicles but fail to
Indian rhinoceros.22 Phytoestrogens have been shown to ovulate in response to either endogenous or exogenous
negatively affect other species by causing cycle irregularity hormonal stimulation. They may become cystic, or they
or predisposing them to reproductive diseases and lesions may eventually luteinize. Anovulatory follicles will often
like pyometra, CEH, and leiomyoma.23,24 prevent a subsequent follicular wave from progressing until
the anovulatory follicle itself regresses. If the anovulatory
Endocrine Disorders follicle does luteinize, prostaglandin administration may
Endocrine disorders of any type should raise suspicion be used to lyse the luteal tissue and stimulate an earlier
that reproductive hormones may also be affected through return to cyclicity by allowing the next follicular wave to
intricately balanced webs of positive and negative feedback progress. The presence and progression of these follicles is
loops. Hyperadrenocorticism, hypothyroidism, hyperthy- best documented by ultrasound.
roidism, and hyperprolactinemia all prevent normal ovarian In polyestrous species, prolonged interestrous intervals
function in females.25,26 Treatment with cabergoline, while may be caused by persistent corpora lutea (CL) that do
lowering prolactin levels, did not result in resumption of not regress after maternal recognition of pregnancy (MRP)
cyclicity in elephants.27 fails to occur. This may happen because of disruption to the
Neoplastic ovarian tumors will cause ovarian dysfunction MRP signal or to the receptors responsible for receiving the
either by causing cycle irregularities or complete shutdown signal. Mechanisms for MRP are species specific, so failure
of the estrous cycle, depending on whether or not hormones of MRP would vary in each species. Early loss of undetected
secreted by the tumor affect the contralateral ovary. The pregnancy after MRP appropriately occurred would also
most common ovarian tumor in most species is the granu- result in persistent CL and prolonged interestrous interval.
losa or granulosa-theca cell tumor (GCT). In some species, For unknown reasons, persistent CL are also known to
this tumor secretes inhibin, which will inhibit follicle- occur in nonmated cycles and will result in a prolonged
stimulating hormone (FSH) secretion and therefore prevent interestrous interval.
128 SE C T I O N 4 Reproduction
26. Brown JL, Walker SL, Moeller T: Comparative endocrinol- 30. te Velde ER, Pearson PL: The variability of female reproductive
ogy of cycling and non-cycling Asian (Elephas maximus) and ageing, Hum Reprod Update 8:141–154, 2002.
African (Loxodonta africana) elephants, Gen Comp Endocrinol 31. Rambags BPB, Krijtenburg PJ, van Drie HF, et al: Numerical
136:360–370, 2004. chromosomal abnormalities in equine embryos produced in vivo
27. Morfeld KA, Ball RL, Brown JL: Recurrence of hyperprolac- and in vitro, Mol Reprod Dev 72:77–87, 2005.
tinemia and continuation of ovarian acyclicity in captive African 32. Lockyear KM, Waddell WT, Goodrowe KL, et al: Retrospective
elephants (Loxodonta africana) treated with cabergoline, J Zoo investigation of captive red wolf reproductive success in relation
Wildl Med 45:569–576, 2014. to age and inbreeding, Zoo Biol 28:214–229, 2009.
28. Ball BA, Almeida J, Conley AJ: Determination of serum 33. Rabon DR, Waddell W: Effects of inbreeding on reproductive
anti-Müllerian hormone concentrations for the diagnosis of success, performance, litter size, and survival in captive red
granulosa-cell tumours in mares, Equine Vet J 45:199–203, 2013. wolves (Canis rufus), Zoo Biol 29:36–49, 2010.
29. Hendriks WK, Colleoni S, Galli C, et al: Maternal age and in
vitro culture affect mitochondrial number and function in equine
oocytes and embryos, Reprod Fertil Dev 27:957–968, 2015.
22
Changes in Reproductive Management
CHERYL ASA
130
CHAPTER 22 Changes in Reproductive Management 131
• BOX 22.1 Association of Zoos and Aquariums For most species in animal breeding programs, full-
Committees and Advisory Groups term gestation or incubation are necessary for the birth
Under the Umbrella of the or hatching of healthy offspring that are able to survive.
Reproductive Management Center In mammals, a component of fertility is a healthy uterus
where a conceptus may implant and be nourished until it
Committees Scientific Advisory Groups
is capable of surviving on its own. The situation may be
Wildlife Conservation Reproduction and Endocrine
similar for viviparous reptile species. The analogous period
Management
for birds and some other oviparous species is of course
Animal Health Nutrition
incubation, which requires appropriate behavior by one
Animal Welfare Behavior
or both parents. The behavior sequence may be influenced
Small Population Management
both by learning (experience) and by the hormonal milieu,
Biomaterials Banking
which itself may be affected by the interactions between
mates (e.g., ring doves8,9).
Although not all species provide parental care for their
young, it is a critical phase for most of those in managed
• BOX 22.2 Basic Components in the breeding programs. Failure to display appropriate care may
Reproductive Process Required for be simply a function of experience; that is, first-time parents
Successful Production of Young are often less successful than those who have already gained
Component experience. Although the hand-raising of young when the
Introduction of male and female parents fail to provide adequate care may be deemed neces-
Compatibility for mating sary for their survival, as adults they too may lack parenting
Fertility of both male and female skills, thus perpetuating the cycle of failure. Parenting
Full gestation or incubation may also be affected by the habitat provided or by social
Appropriate parental care grouping, aspects usually considered of husbandry. Again,
knowledge of natural history of the species may inform care.
Although mate choice, as discussed earlier, is familiar
to many animal managers, cryptic female choice is less
a normal and perhaps critical component of the courtship well known but may affect all stages of the reproductive
sequence in some species. A basic familiarity with natural process following mating.10,11 The term refers to effects on
behavior may provide guidance. conception, gestation/incubation, and parental care that
Mate acceptance may also be enhanced by allowing some may be compromised if a female is forced to mate with a
measure of choice.6 Male–male competition and female nonpreferred male. The female may not actually be con-
choice is part of the reproductive process in many species, yet scious of influencing these processes, but—perhaps through
modern breeding programs seldom provide choice; instead, mechanisms associated with the stress response—sperm
they present only the recommended partner. Recent studies transport may be altered, resources may be shunted from
show that breeding outcomes may be improved by offering eggs or embryos, and care may be withheld after birth or
more than one potential mate.7 Logistics is one of the main hatching. These phenomena have been best described for
challenges to presenting choice, so there is a critical need for birds but are believed to affect other taxa as well. This is
devising and testing such models for a variety of taxa and therefore another reason to consider offering mate choice.
situations. Offering choice need not compromise genetic
management, because options might be limited to potential Limiting Reproduction
partners who also satisfy population goals.
Even if a pair is compatible and successfully mates, con- Fulfilling PMC breeding recommendations entails not only
ception depends on both partners being fertile. Assessing producing offspring from chosen pairs but also preventing
fertility has become practical for many taxa with the avail- reproduction among those that have not received a breed-
ability of endocrine monitoring, ultrasound, and semen col- ing recommendation. Thus comprehensive reproductive
lection techniques. In fact, an increasing number of animal management includes not only enhancing reproduction
programs recommend routine endocrine monitoring, which for some but assessing options for preventing the produc-
may often use fecal samples that are relatively simple to tion of offspring in others. Options include separation of
collect for most species, to determine onset of puberty, males from females, reversible contraception, or even, in
breeding season changes, and even time of ovulation. A some cases, permanent sterilization. Contraception recom-
workshop conducted by the RMC in 2015 developed tools mendations in the United States are provided by the RMC
for assessing fertility in a representative ungulate and carni- (formerly the AZA Wildlife Contraception Center) through
vore, something that could be accomplished for other taxa its website www.stlzoo.org/contraception. In Europe, where
as well, by bringing together participants with the needed the availability of contraceptive products varies, a similar
expertise. Approaches to addressing female infertility are service is provided by the European Group for Zoo Animal
presented in Chapter 21. Contraception through the website www.egzac.org.
132 SE C T I O N 4 Reproduction
Contraception has been used primarily in mammals, paradigm also delays breeding for individual females for as
although it is being considered an option in an increasing long as possible, to lengthen generation time and thus lessen
number of other taxa. Even among mammals, a contracep- the loss of gene diversity over time. These strategies are
tive product that is safe and effective in one taxonomic group designed to meet the goals of genetic population manage-
may present risks in another. The best example is progestin- ment, but they may compromise the fertility of females in
based methods, such as the melengestrol acetate (MGA, the population.
Wildlife Pharmaceuticals) implant and Depo-Provera injec- However, this paradigm may be at odds with reproduc-
tions (medroxyprogesterone acetate: Schering), that have tive biology. In a study of canid species managed in AZA
been effective and safe in most species for decades. Efficacy institutions, the prevalence of endometrial hyperplasia and
is virtually 100% in females of all species treated, but few pyometra was higher in females that had reproduced less
studies of reversibility have yet been conducted (golden lion frequently.21 A review of the literature22 found “use it or
tamarin,12 tiger13) due to the complexity of judging rever- lose it” to be a theme common to a number of species
sal (e.g., mate access and need to compare to individuals (e.g., cheetahs,23,24 elephants,25 white rhinoceroses,26 Seba’s
matched by age and reproductive history).14 RMC staff are bats,27) as hypothesized by Lindburg and Durrant 20 years
conducting reversibility studies of two representative Old ago.28 There are likely taxonomic differences in susceptibil-
World monkey species, colobus and hamadryas baboon, ity to endometrial effects, and other components of the
to determine duration of implant efficacy more specifically reproductive system may be affected differentially across
so as to facilitate subsequent breeding recommendations. species. Infertility resulting from long barren periods is
Surveys submitted to the Contraception Database and avoided in some countries by management euthanasia (see
subsequent investigation by RMC and RHSP staff have Chapter 23).
failed to identify contraceptive failures or significant health A related phenomenon is the importance in many species
effects in the wide range of taxa that have been treated with of establishing fertility in young females by allowing early
the exception of carnivores. Studies have shown that these reproduction. In general, allowing a female to reproduce
same progestin-based products carry a risk of endometrial soon after puberty seems to affect the reproductive process
and mammary pathology in felids15 and canids16; and the in ways that increase fertility in later years, although the
response is likely similar in other carnivore species. Cur- mechanisms may vary by species. However, the concern
rently the best option for carnivores is a GnRH agonist of population geneticists is that this decreases generation
(deslorelin: Suprelorin, Virbac Animal Health) paired with time and risks losing gene diversity. However, reproductive
a short-term progestin to prevent the initial stimulation failure itself results in loss of gene diversity,
phase,17 although time to reversal may vary greatly among
individuals and species (M. Agnew, unpublished). Lifetime Reproductive Planning
The GnRH agonist implant Suprelorin has also been
used successfully in taxa other than carnivores.18 Efficacy in The success of genetic population management rests on rec-
females of most mammalian species is high once the correct ommended breedings resulting in the production of young,
dose has been determined. Although GnRH agonists should but as outlined earlier, reproductive rates are so low that
also be effective in males, higher dosages than those used in most programs are not meeting their goals. Thus strategies
females of the same species are typically needed. However, for establishing and maintaining fertility are crucial. In the
males of most ungulate species have not been successfully United States, the RMC is developing and testing models
suppressed. Use in some species of birds and reptiles has for lifetime reproductive planning for individual females.
increased, but efficacy has been mixed. Whether the out- Breeding recommendations would not be based only on
comes are due to species differences or inadequate doses is current factors related to population genetics. Instead, the
yet unknown. potential genetic contribution of each female to the popula-
Although not specifically reproductive management, tion could be developed (i.e., planned soon after her birth
euthanasia of individual animals judged genetically surplus and before puberty). The ideal pattern for most species
to the population is an option in some countries and is would allow each female to reproduce soon after reaching
being increasingly discussed in the United States.19,20 See puberty, to establish fertility, and then at regular intervals
also Chapter 23. to maintain fertility: “Breed early and often.”
Various scenarios are being modeled, an analysis that
must be repeated for each species owing to differences in
Genetic Management Versus parameters such as mating system, ages of first and last
Reproductive Management reproduction for the population, interbirth or interclutch
intervals, litter or clutch sizes, and so on. For some females,
In current practice, breeding recommendations are gener- the Lifetime Reproductive Planning might be to produce
ated for most species once annually or at least every few their entire lifetime contribution early in life without inter-
years based on the genetic composition at the time of ruption. For others, breeding recommendations would be
analysis. That paradigm results in some females not receiv- spread across a lifetime or concentrated during prime breed-
ing a recommendation to reproduce for many years. The ing age, usually middle age. During nonbreeding intervals,
CHAPTER 22 Changes in Reproductive Management 133
these females would be either separated from males or 10. Eberhard WG: Female control: sexual selection by cryptic female
treated with the contraceptive that is most appropriate for choice, Princeton, NJ, 1996, Princeton University Press.
the species in terms of efficacy, safety, and reversibility. Once 11. Zeh JA, Zeh DW: The evolution of polyandry. II. Post-
copulatory defenses against genetic incompatibility, Proc Royal
a female’s genetic contribution to the population is secure,
Soc B 264:69–75, 1997.
she could be permanently sterilized or given a long-term 12. Wood C, Ballou JD, Houle CS: Restoration of reproductive
contraceptive, which could retain the possibility of later potential following expiration or removal of melengestrol acetate
reproduction if changes in the population warranted. contraceptive implants in golden lion tamarins (Leontopithecus
Lifetime reproductive planning is proposed as a step rosalia), J Zoo Wildl Med 32:417–425, 2001.
in addressing reproductive failure in mammals, caused in 13. Chuei JY, Asa CS, Hall-Woods M, et al: Restoration of repro-
particular by endometrial disease resulting from long barren ductive potential after expiration or removal of melengestrol
periods. This appears to be a common problem in some acetate contraceptive implants in tigers (Panthera tigris), Zoo Biol
mammalian species, but other causes of reproductive failure 26:275–288, 2007.
must be considered as well. Especially in nonmammalian 14. Asa CS: Assessing efficacy and reversibility. In Asa CS, Porton
taxa, adjusting husbandry protocols may be pivotal, and, IJ, editors: Wildlife contraception: issues, methods and application,
Baltimore, 2005, Johns Hopkins University Press, pp 53–65.
for all species, behavioral sequences specific to courtship
15. Munson L: Contraception in felids, Theriogenology 66:126–134,
and social interactions should be considered. Similarly, 2006.
overall health is central to fertility and the production of 16. Moresco A, Munson L, Gardner IA: Naturally occurring and
young that survive. For example, veterinarians may play melengestrol acetate-associated reproductive tract lesions in zoo
a more central role in enhancing reproductive success canids, Vet Pathol 46:1117–1128, 2009.
by incorporating breeding soundness exams as a routine 17. Wright PJ, Verstegen JP, Onclin K, et al: Suppression of the
protocol—something that is also recommended by the AZA oestrous responses of bitches to the GnRH analogue deslorelin
Animal Health Committee. by progestin, J Reprod Fertil Suppl 57:263–268, 2001.
18. Agnew M, Asa CS: A summary of Suprelorin use in United States
References zoos: taxonomic scope, sex differences, and efficacy, J Zoo Wildl
Med submitted.
1. Baker A: Animal ambassadors: an analysis of the effectiveness and 19. Powell DM, Ardaiolo M. Survey of U.S. zoo and aquarium
conservation impact of ex situ breeding efforts. In Zimmermann animal care staff attitudes regarding humane euthanasia for
A, Hatchwell M, Dickie L, et al, editors: Zoos in the 21st century: population management, Zoo Biol 35:187–200, 2016.
Catalysts for conservation? London, 2007, Zoological Society of 20. Asa CS: Weighing the options for limiting surplus animals, Zoo
London, pp 139–154. Biol 35:183–186, 2016.
2. Lees CM, Wilcken J: Sustaining the Ark: the challenges faced by 21. Asa CS, Bauman KL, Devery S, et al: Factors associated with
zoos in maintaining viable populations, Intl Zoo Yrbk 43:6–18, uterine endometrial hyperplasia and pyometra in wild canids:
2009. implications for fertility, Zoo Biol 33:8–19, 2014.
3. Lacy R: Achieving true sustainability of zoo populations, Zoo Biol 22. Penfold L, Powell D, Traylor-Holzer K, et al: “Use it or lose it”:
32:19–26, 2013. characterization, implications, and mitigation of female infertil-
4. Lacy RC: Managing genetic diversity in captive populations of ity in captive wildlife, Zoo Biol 33:20–28, 2014.
animals. In Bowles ML, Whelan CJ, editors: Restoration and 23. Hermes R, Hildebrandt TB, Göritz F: Reproductive problems
recovery of endangered plants and animals, Cambridge, 1994, directly attributable to long-term captivity – asymmetric repro-
Cambridge University Press, pp 63–89. ductive ageing, Anim Reprod Sci 82–83:49–60, 2004.
5. Ballou JD, Foose TJ: Demographic and genetic management of 24. Crosier AE, Comizzoli P, Baker T, et al: Increasing age influences
captive populations. In Kleiman DG, Lumpkin S, Allen M, et al, uterine integrity, but not ovarian function or oocyte quality,
editors: Wild mammals in captivity, Chicago, 1996, University of in the cheetah (Acinonyx jubatus), Biol Reprod 85:243–253,
Chicago Press, pp 263–283. 2011.
6. Asa CS, Traylor-Holzer K, Lacy R: Can conservation-breeding 25. Agnew DW, Munson L, Ramsey EC: Cystic endometrial hyper-
programmes be improved by incorporating mate choice?, Intl Zoo plasia in elephants, Vet Pathol 41:179–183, 2004.
Yrbk 45:203–212, 2011. 26. Hermes R, Hildebrandt TB, Waltzer C, et al: The effect of long
7. Martin MS, Shepherdson DJ: Role of familiarity and preference non-reproductive periods on the genital health in captive female
in reproductive success in ex situ breeding programs, Conserv Biol white rhinoceroses (Ceratotherium simum simum, C.s. cottoni),
26:649–656, 2012. Theriogenology 65:1492–1515, 2006.
8. Cheng MF: Female cooing promoter ovarian development in 27. Napier JE, Caron S, Reavill DR, et al: Proliferative endometrial
ring doves, Physiol Behav 37:371–374, 1986. lesions in a group of Seba’s short-tailed bats (Carollia perspicil-
9. Kroodsma DE: Reproductive development in a female song- lata), J Zoo Wildl Med 40:437–444, 2009.
bird: differential stimulation by quality of male song, Science 28. Lindburg D, Durrant B: Delayed reproduction: the good and the
192:574–575, 1976. bad, Zoo Biol 17:369–371, 1998.
23
Issues Surrounding Surplus
Animals in Zoos
MADS FROST BERTELSEN
“S
urplus animal” has a negative connotation, and mate choice, increase genetic diversity, and address infertility
it seems appropriate to start a discussion around (see Chapter 21). That is, a reproductive management plan
this topic with a definition. Surplus animals are must be implemented for each individual to optimize the
surplus to the needs of the population and in excess of possibility of retaining genetic diversity through breeding
the needs of the individual institution. In other words, a while maintaining manageable yet sustainable populations.
surplus is more likely to occur the better a species is doing
in zoos. The more offspring that are born and the better the Meeting the Demand
individuals are doing in terms of coping with disease, stress,
and other problems, the more likely it is that the supply Contraception or even just separation of the sexes are
will exceed the demand. Fundamentally surplus animals are powerful tools to reduce the number of offspring. However,
a sign of success. The day when zoos breed a surplus of all the safety and reversibility in terms of future breeding
endangered animals would be a day to celebrate. However, are often (depending on the species) less than optimal.6,7
surplus animals eat, take up space (which is ultimately More importantly though, sustainability is not just about
always limited), and evoke the emotions of staff and visitors, numbers but about breeding the right animals. Although
so their management is a complicated issue. sometimes skewed,8 the average sex ratio at birth is close to
1:1, producing an unavoidable surplus of males in species
Sustainable Populations where one male breeds with several females (e.g., a “harem”
134
CHAPTER 23 Issues Surrounding Surplus Animals in Zoos 135
Breeding Is “Natural”
meat,12 and every zoo utilizes invertebrates, rodents, chick-
In general, zoos strive to provide “natural” conditions for ens, and ungulates as feed for its carnivorous inhabitants. In
their animals, although in practice numerous compromises addition, most zoos kill invertebrates, rodents, and various
are made; “natural” space is not available to most animals, other animals categorized as pest species. An old anecdote
“natural” diets are often substituted, and “natural” habitats accounts for a conversation between a gentleman and a
and climates are mostly lacking. On the upside, “natural” distinguished lady at a fundraising dinner. The gentleman
parasites, “natural” predator stress, and “natural” competition offers the lady $100,000 if she will agree to sleep with him,
for food are usually absent. Most would agree that “natural” an offer to which she assents. He then asks if she would do it
behavior should be strived for, and with food provided for $10. The lady gets upset and says: “What kind of woman
and no predators to avoid, breeding becomes a paramount do you think that I am?” to which he replies: “We have
tool in providing “natural” behavior and “enrichment” in already established that. Now we are just haggling over the
the shape of courtship, pair bonding, mating, pregnancy, price.” The situation is very analogous to our relationship
nursing, feeding, mother–infant bonding, playing, sparring, to killing animals; consciously or not, we all apply a more
and so on.9–11 All these effects are essential parts of animal or less arbitrary cutoff on a scale from cockroach to great
welfare, but in excess of population needs, surplus animals ape (Fig. 23.2), and our position on the scale is highly
are the unavoidable secondary outcome. dependent on our nationality and cultural background.10,13
When the rational decision to cull has been made, the next
How to Deal With Surplus Animals question is when to do so. Some institutions have instituted
a practice of culling infants deemed surplus shortly after
So for the reasons previously mentioned, a certain surplus birth; however, this precludes them from harvesting several
of animals is not only a sign of healthy populations but of the benefits of producing surplus animals: the enormous
also an unavoidable “by-product” of sustainable breeding. behavioral enrichment to the parents of raising the offspring
As previously mentioned, simply housing surplus animals and the idea of having a buffer. A compromise, based on
indefinitely is counterproductive to achieving sustainable the three peaks of mortality observed in the wild, appears
populations, as these animals take up space that could rational and “natural”: In “nature” the mortality is highest
be used for individuals more genetically valuable to the in infants, animals around dispersal age, and in animals past
population. Sending such animals to private holders or their prime; geriatrics are not common in the wild. Zoos
institutions outside of the breeding programs raises a mul- can mimic this by reducing litter sizes perinatally, primar-
titude of ethical issues and ultimately is not a long-term ily culling around dispersal age, and by minimizing the
solution. Reintroduction into the wild unfortunately is amount of postreproductive animals to a minimum deemed
rarely a realistic solution. Thus the only option available is necessary for balanced group composition. Maintaining
to kill (or cull) those animals definitely in surplus. postreproductive individuals of solitary or monogamous
It can be (and has been) argued that killing any animal species is counterproductive for population sustainability.
is ethically wrong; however, the vast majority of human How the animals are used following culling has a great
beings and every zoo known to the author have made the impact on the acceptance of the practice by zoo employees
fundamental choice that it is acceptable to kill animals. For and the public alike. Also here, there are vast cultural dif-
example, approximately 95% of the US population consume ferences around the globe, yet it appears that a utilitarian
136 SE C T I O N 4 Reproduction
Therapeutics
24 Stem Cell Therapy in Zoo Medicine, 138
137
24
Stem Cell Therapy in Zoo Medicine
MATTHEW E. KINNEY AND ROBERT HARMAN
What Are Stem Cells? A 2006 position statement from the international society
for cellular therapy sought to clarify the definition in an
Stem cells are defined as having both the capacity for attempt to better characterize MSCs and allow for improved
self-renewal and the ability to give rise to differentiated standardization in the rapidly evolving field of stem cell
cells.1–4 Stem cells are classified based on their differentia- therapy and regenerative medicine.13 The minimum criteria
tion potential and cell origin. Cells have diverse abilities to for defining a human MSC were based on the ability to
differentiate and can be characterized based on a continuum adhere to plastic, specific surface antigen expression, and
from unipotent cells that are capable of differentiating into multipotent differentiation potential. Plastic adherence is
a single mature cell type to totipotent cells that are capable a common nonspecific but well-described characteristic of
of differentiating into any cell or tissue of an organism. MSCs under standard culture conditions. Immunopheno-
Classification based on cell origin is used to distinguish typing using expression (CD105, CD72, CD90) and lack
embryonic stem cells (ESCs) from adult tissue–derived of expression (CD45, CD34, CD14 or CD11b, CD79
stem cells. ESCs arise from embryos during the early alpha, or CD19, HLA-DR) of specific surface antigens is
stages of development. Cells of the inner cell mass are able used as a proxy to identify MSCs and make certain that
to generate into any cell type of the body and maintain contamination from other cells, such as hematopoietic cells,
unlimited proliferation and replication under appropriate has not occurred. In human medicine using flow cytometry,
culture conditions.5 In 1998 the establishment of human 95% or more of MSCs should be positive for the specific
ESC lines was first reported, and although the research antigens and less than 2% of MSCs should lack expression
potential of a cell that can proliferate indefinitely into of the specific antigens listed previously. The use of these
any cell type of the body is enormous, the clinical use of specific surface antigens to discriminate MSCs from other
ESCs in veterinary medicine as a therapeutic modality has cells types cannot be universally translated from human to
minimal direct utilizations at this time due to the high risk veterinary medicine, as there are clear interspecies differences
of tumors.6,7 As such, the focus of this chapter is on adult in the expression of surface antigens.14 The third criterion
tissue–derived stem cells, which are currently being utilized is based on cell potency and states that using standard
in therapeutic capacities in both human and veterinary in vitro culture conditions, MSCs should demonstrate the
medicine. Induced pluripotent stem cells (iPSCs) are adult ability to differentiate into all three mesenchymal lineages,
cells that are reprogrammed to generate cell lines that have including osteoblasts, adipocytes, and chondroblasts. Dif-
the capacity for self-renewal and pluripotency. The use of ferentiation into cells of the mesoderm is merely a minimal
iPSCs for conserving and rescuing endangered species is an criterion for definition, as some authors contend that
exciting and promising use of iPSCs in veterinary medicine, MSCs have the capacity to differentiate into cell lineages
along with many other research applications; but iPSCs have other than those of mesodermal origin.15 In 2013 a similar
many of the drawbacks of ESCs, including immunogenicity position statement was proposed under the authority of
and tumorogenicity.7–11 Mesenchymal stem cells (MSCs) are the International Federation for Adipose Therapeutics and
another type of adult-derived stem cells and are commonly Science and the International Society for Cellular Therapy
used as a therapeutic modality in veterinary medicine. The to provide guidance and establish standards for researchers
reader should be aware that MSC, an abbreviation present investigating adipose-derived cells for use in regenerative
throughout the stem cell literature, may be a reduction medicine.16
for any one of the following terms; mesenchymal stem cell, MSCs are an attractive therapeutic modality in veterinary
mesenchymal stromal cell, multipotent stromal cell, marrow medicine because these cells are multipotent, present in a
stromal cell, colony forming fibroblasts, or medicinal signaling number of accessible tissues for cell collection, have the
cell. The naming convention for MSCs was first based on capacity for self-renewal, can expand using established in
their ability to differentiate into cells of mesodermal origin.12 vitro culture methods, are genetically stable, have powerful
138
CHAPTER 24 Stem Cell Therapy in Zoo Medicine 139
trophic effects, and have few ethical concerns.17,18 For these factors produced by stem cells may have therapeutic poten-
reasons MSCs are the most commonly utilized stem cell tial without administration of actual cells to the patient.
type for therapeutic purposes in veterinary medicine and
a number of both university and commercial research Stem Cell Collection
groups continue to facilitate the progress of this modality
in research and clinical veterinary medicine. Autologous is a term used to describe MSCs harvested from
and administered to the same individual. An allogeneic rela-
How Do Stem Cells Work? tionship between MSC donor and recipient indicates that
both are from the same species but different individuals. A
The mechanism of MSC function is an active and rapidly xenogeneic relationship indicates the MSC donor and recipi-
evolving area of research, and many questions remain to be ent are from different species. Currently the most common
elucidated in determining how MSCs behave both in vitro donor-recipient relationship in veterinary medicine is
and in vivo. The original hypothesis that MSCs exhibit autologous, although allogeneic MSC administration has
their therapeutic effect by direct tissue regeneration was been performed in a number of species. The use of alloge-
logical and intuitive based on their ability for self-renewal neic MSCs in veterinary medicine can potentially provide
and multipotent differentiation in vitro.19 The capacity consistency and standardization of MSCs administered
of MSCs to migrate to the site of injury, engraft, and to the recipient and allow for immediate administration
differentiate into functional cells was initially postulated to a recipient without the need to harvest tissue prior to
to be the underlying mechanism of clinical improvement MSC administration. Studies evaluating allogeneic MSC
observed following MSC administration; perhaps because administration by various routes have reported few adverse
of the relative simplicity, this viewpoint remains a popular clinical concerns in canine or equine patients, and the com-
understanding. However, the paradigm of direct tissue mercialization of allogeneic MSC is a promising research
regeneration secondary to MSC migration and incorpora- direction that holds great potential for zoo medicine.30–32
tion has been questioned based on clinical improvement MSCs are currently thought to reside in large part in
observed despite the lack of long-term stem cell engraftment a perivascular niche and have been termed pericytes.33,34
in target tissues and numerous studies proposing additional Although MSCs reside in nearly all organs, the feasibility of
mechanisms of action. The mechanism of action of MSCs is cell collection is an essential clinical consideration in deter-
likely a combination of differentiation into functional cells, mining a suitable harvest location. In human and veterinary
bioactive factor secretion, cell-to-cell interactions, and the medicine, bone marrow, adipose tissue, and umbilical tissue
release of extracellular vesicles that are dependent on MSC are the most common locations for MSC harvesting. The
origin, culture condition, administration protocols, local optimal tissue source of exogenous MSCs has not been
microenvironments, and desired therapeutic response.20–22 determined.17 Bone marrow MSCs (BM-MSCs) were the
Bioactive factor secretion is intended to be a phrase that original MSCs isolated; collection protocols from the fifth
encompasses a variety of terms in the stem cell literature sternebra have been thoroughly described in sedated horses
documenting the ability of MSCs to affect other cells. using local anesthetic and a 12-gauge Jamshidi needle.35,36
This includes autocrine, paracrine, and stem cell–related The fifth sternebra is the preferred collection site because it
molecules as well as trophic factors. Considerable research, is cranial to the apex of the heart, and the marrow space is
particularly in the area of myocardial and central nervous of adequate size to permit the harvesting of 10–15 mL of
system ischemic insult, has focused on the investigation bone marrow. Adipose tissue is an attractive harvest location
of cytokines, growth factors, chemokines, and immu- due to its abundance and relative ease of collection. Adipose
nomodulatory proteins and how they contribute to the MSCs (AT-MSCs) have been harvested from both periph-
therapeutic improvements observed following stem cell eral and visceral adipose tissue, with the most common
administration.23 This area of stem cell research in humans location in the horse being the subcutaneous tissue near
and laboratory animals has focused on characterizing the the tail head or over the dorsal gluteal muscles. Similarly,
transcriptome and proteome in an effort to explain the a lipectomy performed at the base of the tail in a domestic
role played by MSCs in immunomodulation, decreasing cow and water buffalo (Bubalus bubalis) has been used to
apoptosis, promoting cell survival, accelerating progenitor collect and culture AT-MSCs, and this collection location
cell self-renewal, stimulating angiogenesis, blocking pain, is suitable for many zoo species.37 In the bottlenose dolphin
and reducing inflammation.24–29 Numerous complete (Tursiops truncatus), adipose-derived MSCs were harvested
reviews are available detailing the proposed underlying and subsequently cultured using ultrasound-guided lipo-
mechanisms, but the crucial take away for the veterinary suction from the subcutaneous fat on the dorsal midline,
clinician is to recognize that the mechanism of action of 15 cm caudal to the blowhole, following local anesthesia
MSCs is not to merely engraft into diseased tissues for and use of a 2.4-mm diameter infusion cannula.38 As
replacement but rather influence the microenvironment to compared with subcutaneous fat, blubber contained
promote functional recovery of an organ or body system.19–27 fewer nucleated cells per gram and was difficult to digest
In fact, it is feasible that under appropriate harvest, culture, efficiently. The umbilical cord is another attractive harvest
and expansion conditions, the administration of bioactive location because such harvesting is entirely noninvasive
140 SE C T I O N 5 Therapeutics
and cells are collected from a young tissue source. MSCs intravenous injection, as the cells will traverse to the lung
can be obtained from umbilical cord blood or Wharton capillary beds. Recently, it was reported that MSC admin-
jelly, a collagen-rich matrix within the umbilical cord. The istration to horses with exercise-induced pulmonary hemor-
umbilical cord should be considered as a potential harvest rhage was efficacious in preventing bleeding during racing
location for MSCs in zoological medicine. MSC cultures and could be considered for other respiratory diseases.45
from the umbilical cord of a greater one-horned rhinoc- Knowledge gaps in the area of MSC homing, an active area
eros (Rhinoceros unicornis), giraffe (Giraffa sp.), and lion of research, include the underlying mechanism of homing,
(Panthera leo), among other species, have been established. the activity of MSCs once target tissue has been reached,
In many species, especially megavertebrates, placental and the ability to improve homing by promoting receptor
and umbilical cord tissue collection can be incorporated conservation in MSC cultures. As these knowledge gaps are
into the birth plan of individual animals to streamline filled, systemic MSC administration will likely be modified
the collection process during the periparturient time. The to target damaged tissues and diseases in individual patients,
isolation and culture of MSCs from peripheral blood have with fewer MSCs required to attain the desired therapeutic
been identified as an alternative and attractive source of outcome.46
MSCs owing to the ease of collection; this is an attractive
option for zoo species. The demonstration of chondrogenic Stem Cells in Clinical Medicine
differentiation of human peripheral blood–derived MSCs
suggests that cells derived from peripheral blood may have The therapeutic use of MSCs in veterinary medicine has
similar potential as MSCs derived from adipose tissue, bone been best studied in lameness conditions in canine and
marrow, or umbilical cord.39 Similarly, immunophenotyp- equine patients because of the original postulated mecha-
ing and histochemical staining of MSCs isolated from nism of tissue regeneration and the interest in exploring this
equine peripheral blood supports the potential for trilineage emerging technology as an adjunctive treatment modality
differentiation.40 MSCs have been isolated and cultured complementary to or in place of more traditional treat-
from the peripheral blood of a giraffe (Giraffa spp.), ments for musculoskeletal injuries. The efficacy of MSC
further supporting the potential use of this collection site treatment in domestic dogs has progressed from anecdotal
in zoo species (Valerie Johnson, personal communication, reports of clinical improvement to prospective, randomized,
March 2, 2017). placebo-controlled clinical studies with a relatively large
Intravenous, intraarterial, intraperitoneal, and local number of study participants.32 Outcome metrics have
intratissue routes have been used to deliver MSCs to human evolved from owner-reported improvement to the utiliza-
and veterinary patients. The size of some zoo species must tion of force platform gait analysis, radiographs, synovial
be considered in determining an administration route, fluid analysis, client-specific outcome measurements using
as intratissue administration may be precluded by organ a standardized scoring system, and a scoring system gauging
accessibility or the need for restraint. Homing is a term veterinary pain on manipulation.32,47–50 The refinements in
used to describe the arrest of MSCs within the vascula- study design and outcome metrics provide valuable safety
ture, followed by transmigration across the endothelium; and efficacy data supporting the potential applications of
it is postulated that injured or inflamed tissues release MSCs for musculoskeletal conditions, and further studies
chemokines, cytokines, and growth factors that serve as should continue to utilize blinded, randomized, placebo
a cue to direct MSCs to tissues in need of repair.41 In controlled studies to allow clinicians to practice evidence-
general, MSC homing to target tissues occurs when there based medicine when considering MSCs as a potential
is the right combination of signaling molecules from the treatment modality.
injured tissue and the corresponding receptors on MSCs.42 Increased understanding of the mechanisms underlying
Rats with myocardial infarctions administered intravenous MSC activity has shifted the fundamental perception of
BM-MSCs in the ascending aorta demonstrated MSC their usefulness in veterinary medicine from a therapy based
homing to the infarcted regions of the heart and improved on tissue-regenerative properties to one that may be utilized
fractional shortening.43 Increased MSC engraftment in the to treat a variety of diseases. MSC-based therapies for the
gastrointestinal tract, liver, and spleen was documented in treatment of equine joint disease, tendon and ligament
mice that received local radiation to the abdomen when injuries, and bone repair were an early use and continue
compared to nonirradiated mice, suggesting that MSC today.51–55 In the domestic horse, equine recurrent uveitis,
homing occurs in radioinduced lesions.44 The concept laminitis, perinatal asphyxia syndrome, chronic corneal
of MSC homing is promising in zoological medicine, as ulceration, equine myeloencephalopathy, and laryngeal
this characteristic of MSCs may be harnessed to facilitate hemiplegia have been specifically identified as conditions
the targeting of damaged tissue via systemic intravascular that may benefit from MSC treatment.56 Veterinary der-
administration. In instances where the damaged tissue is matology has been identified as a specialty where MSCs
extensive and diffuse or the patient is too large to facilitate may expand the treatment options for chronic nonhealing
local MSC administration, systemic administration may be wounds, immune-mediate skin diseases, alopecia, and scar
more practical and efficacious compared with intralesional tissue remodeling based on the antiinflammatory, immuno-
injection. MSC therapy of the lung can be easily applied via modulatory, revascularization, and antiapoptosis effects as
CHAPTER 24 Stem Cell Therapy in Zoo Medicine 141
well as the differentiating and homing capacities of MSCs.57 cells do not appear to be the sole explanation for clinical
Domestic cats refractory to conventional treatment for improvement. The paracrine effects of MSCs are suspected
feline chronic gingivostomatitis were administered two to contribute to the cardiac repair by neovascularization,
systemic intravenous injections of adipose-derived MSCs 1 angiogenesis, decreasing inflammation and infarct size,
month apart with oral biopsies, blood immune cell subsets, enhancing cardiomyocyte survival, mobilizing other stem
serum protein, and cytokine levels used as outcome metrics. cells to the infarcted area, and decreasing fibrosis. Similar to
Of the 7 cats that completed treatment, 3 demonstrated myocardial infarction, ischemic stroke has been extensively
complete clinical remission, and 2 showed substantial researched as a condition that may benefit from MSC treat-
clinical improvement. Systemic immunomodulation was ment, with similar variability in cell type, mode of delivery,
documented in cats that responded to treatment with and outcome metrics. Initial studies have reported improved
decreased circulating CD8+ T cells, decreased neutrophil functional recovery, reduced infarct volume, and dimin-
counts, and normalization of CD4/CD8 ratios, among ished neurobehavioral deficits in MSC-treated patients.61
other changes in cytokine concentrations.58 In domestic For both cardiac and cerebral ischemic conditions, rigorous
dogs and cats, MSC treatment has also been attempted for investigation of the efficacy of MSCs will continue due to
a variety of disease conditions, including intervertebral disk the lack of current treatments that offer cures instead of
disease, atopic dermatitis, perineal fistulas, inflammatory merely clinical compensation. MSCs have been identified as
bowel diseases, dilated cardiomyopathy, keratoconjunctivits an innovative treatment option in a number of other disease
sicca, granulomatous meningomyeloencephalitis, and feline processes that affect humans; current studies are evaluating
chronic kidney disease.59 In many of these studies, the safety MSC treatment for acute kidney injury, chronic kidney
of MSC was established; however, efficacy was difficult disease, diabetic nephropathy, glomerulosclerosis, kidney
to evaluate due to study design; it was difficult to assess transplantation, retinal degeneration, glaucoma, diabetes
outcome metrics and follow-up of study patients. In many mellitus, and liver fibrosis, among many other disease
instances the shortcomings of these studies are intrinsic processes, thus highlighting the diversity of MSCs treatment
to the challenges associated with evaluating the efficacy of potential.62–65
a treatment modality on client-owned animals that have
attempted other treatments, may have additional comor- Mesenchymal Stem Cells in Zoo Medicine
bidities, and may elect to no longer enroll in the study or
to euthanize their pets. Nonetheless, these studies provide Despite the paucity of refereed publications on MSCs in
a foundation for further investigations and demonstrate zoological medicine, MSCs have been used in multiple
the potential of MSCs as a treatment modality beyond the zoos. Some examples include the use of intravenous or
regenerative model in domestic animal medicine. intralesional autologous adipose MSC administration for
The use of MSCs in human and research animal medi- the treatment of lameness in a variety of large cats, the
cine provides an additional resource for veterinarians to northern white rhino (Ceratotherium simum cottoni), giraffes
consider when evaluating MSC safety and efficacy. Similar (Giraffa spp.), and Grevy’s zebra (Equus grevyi). Allogeneic
to domestic animal medicine, the preliminary view of the adipose MSCs administered intravenously and topically
regenerative capacity of MSCs resulted in investigation of have also been used to treat pododermatitis and osteoarthri-
MSCs for treatment of diseases that affected body systems tis in macaroni (Eudyptes chrysolophus), emperor (Apteno-
with suspected minimal regenerative capacity. Numer- dytes forsteri), Adelie (Pygoscelis adeliae), gentoo (Pygoscelis
ous human studies of MSC administration following papua), and magellanic (Spheniscus magellanicus) penguins.
myocardial infarction have been performed with different Numerous additional treatments have likely occurred in
cell types, modes of delivery, and clinical outcomes that zoos around the world that have not been widely reported.
contribute to continued debate regarding the benefits of The novelty of MSC treatment often attracts media cover-
MSCs.60 Briefly, AT-MSCs and BM-MSCs are the most age, but unfortunately these cases rarely transition into
commonly administered types with modes of delivery case reports or case series that are presented at zoological
including peripheral intravenous and direct intramyocar- medicine conferences or peer-reviewed publications. There-
dial injection during coronary artery bypass graft surgery, fore MSC harvest sites, culture conditions, administration
transendocardial injection, and intracoronary infusion routes, adverse effects, outcome metrics, and long-term
into the infarcted artery with or without direct adventi- efficacy are largely unknown at this time. Documentation
tial delivery. Several studies have documented improved of MSC use in zoo species will serve not only to introduce
ejection fraction, wall motion, and infarct size in patients basic collection and administration methods but also allow
with acute myocardial infarction and decreased mortality readers to critically evaluate MSCs as a potential treat-
in patients in heart failure, while others have documented ment modality. As the available literature increases, harvest
no significant difference in outcome measurements when locations, administration routes and doses, outcome expec-
compared with conventional treatment. The mechanisms tations, and safety concerns will become more apparent,
underlying these improvements continue to be discussed, allowing clinicians to better evaluate whether MSC treat-
and mechanisms other than the direct differentiation of ment should be pursued as a treatment option in zoological
MSCs into functioning cardiomyocytes replacing necrotic medicine.
142 SE C T I O N 5 Therapeutics
When compared with domestic animal veterinary number of species through the collection of umbilical tissue
medicine, MSC use in zoological species presents a number following parturition or adipose tissue collection during
of unique challenges. Some of these challenges are simply immobilization procedures. Although the initial investment
technical and may be overcome with experience, commu- in establishing and maintaining MSCs is significant, the
nication, and creativity. Determination of the MSC stem potential for rapid turnaround time between identifying a
cell harvest location is an initial challenge. A combination potential case where MSCs may be indicated and treatment
of necropsy photo review, preanesthesia ultrasonography, initiation will allow for the utilization of this modality more
and review of the equine literature provided adequate willingly and rapidly. The potential for the allogeneic or
background knowledge to guide adipose tissue harvest from xenogeneic use of MSC is appealing, but it must be noted
a northern white rhino. In many instances two immobiliza- that aside from select species, the safety of allogeneic admin-
tions are required to harvest fat or bone marrow and to istration has not been elucidated, and there are also logistic
administer autologous MSCs to the patient. The legitimacy considerations such as time, labor, financial investment,
of this concern may be institution-dependent, and repeat the selection of donor animals, ownership of biological
immobilizations are not always required. Currently there material, and accountability for adverse effects, especially if
are commercial companies that will perform patient-side multiple institutions are collaborating on the establishment
harvesting and administration under the same anesthetic of a central repository.
event in domestic animals and the selected administration As the underlying mechanisms of MSCs continue to
route, such as intravenous, may preclude the need for a be better understood, the potential indications for MSCs
second anesthesia in a well-trained patient. Additionally, the have evolved from being a last ditch therapeutic modality
culturing and expansion of MSCs from peripheral blood in for musculoskeletal conditions to a treatment option to
the horse provides a foundation to establish techniques that address a range of diseases in a number of body systems. As
would allow for MSCs to be expanded in adequate numbers our understanding of the mechanisms of MSCs continues
from a biological source that could be voluntarily acquired to advance, the indications for their potential use expand,
from many zoological species including megavertebrates in and many of the basic principles of MSC therapy may
modern zoos.40 Intravenous administration of allogeneic provide a potential treatment for diseases that affect zoo
MSCs harvested from the peripheral blood of domestic animals. In particular, as the mechanisms of homing are
horses involved 291 domestic horses, and no adverse more thoroughly investigated and the efficacy of intrave-
clinical effects were noted in the 1 year post administration nous treatments is reported, intravascular administration of
monitoring period.66 Similarly, intraarticular administra- MSCs is becoming a more attractive treatment option for
tion of allogeneic MSCs harvested from the adipose tissue zoo patients due to their large size or conditions that involve
of donor domestic dogs were demonstrated to be both safe entire organ systems. The antifibrotic and proangiogenic
and effective compared with saline treatment.32 The result of properties may have utility as a treatment option for great
these studies holds great potential application to zoological ape fibrotic cardiomyopathy or cheetah veno-occlusive
medicine. If the safety of allogeneic MSC administration is disease. The regenerative potential of MSCs may present
translated for zoo species, there is potential that peripheral an alternative option for nonhealing cutaneous wounds in
blood or adipose tissue could be used as a MSC source for columbiformes, pododermatitis in penguins, or postintuba-
other individuals of the same species, thus greatly reducing tion tracheal stenosis in long-necked birds. The immuno-
the investment in MSC acquisition from individual animals modulary effects may be useful as a potential therapy for
that may not be conditioned for voluntary blood collection amyloidosis in bongo (Tragelaphus eurycerus) or alopecia in
or be amenable to immobilization to harvest adipose or Andean (spectacled) bears (Tremarctos ornatus). Veterinary
bone marrow. The use of allogeneic or xenogeneic MSCs clinicians are encouraged to consider the potential of MSCs
is a fascinating and promising prospective direction that beyond simply their cell regenerative capacity. Clearly MSC
zoos may consider to separate the source of MSCs from the use as a therapeutic modality in zoological medicine is in
animal that is in need of treatment. The use of allogeneic its infancy, and fundamental questions regarding safety
MSCs could better capitalize on the potential of MSCs and logistics remain unanswered. Basic science research,
by making this treatment modality more similar to other domestic animal and human clinical trials, and veterinary
therapeutics that are immediately initiated once they are colleagues at other zoos may be consulted to expand our
indicated for treatment by an attending veterinarian. The understanding of the underlying mechanisms of MSCs,
utilization of allogeneic or xenogeneic MSCs would allow provide a foundation for translation into zoological medi-
for rapid turnaround time between a veterinary clinician cine, and broaden the potential applications of MSC use
deciding that MSCs are indicated as a treatment option and as a therapeutic modality for various disease processes to
administration to the patient. Currently the San Diego Zoo improve the health of animals in our care.
Safari Park maintains MSC cultures that were established
from adipose or umbilical tissue from northern white References
rhinoceros (C. s. cottoni), greater one-horned rhinoceros
(R. unicornis), Ugandan giraffe (Giraffa camelopardalis 1. Till JE, McCulloch EA: Hematopoietic stem cell differentiation,
spp.), and African lion (P. leo), with plans to increase the Biochim Biophys Acta 605(4):431–459, 1980.
CHAPTER 24 Stem Cell Therapy in Zoo Medicine 143
2. Morrison SJ, Shah NM, Anderson DJ: Regulatory mechanisms 23. Stewart MC, Stewart AA: Mesenchymal stem cells: character-
in stem cell biology, Cell 88(3):287–298, 1997. istics, sources, and mechanisms of action, Vet Clin North Am
3. Weissman IL: Stem cells: units of development, units of regenera- Equine Pract 27(2):243–261, 2011.
tion, and units in evolution, Cell 100(1):157–168, 2000. 24. Glenn JD, Whartenby KA: Mesenchymal stem cells: Emerging
4. Ramalho-Santos M, Willenbring H: On the origin of the term mechanisms of immunomodulation and therapy, World J Stem
“stem cell”, Cell Stem Cell 1(1):35–38, 2007. Cells 6(5):526–539, 2014.
5. National Institute of Health Web site: Embryonic Stem Cells. 25. Billing AM, Ben Hamidane H, Dib SS, et al: Comprehensive
Available at: Yu J, Thomson JA. Embryonic Stem Cells: https:// transcriptomic and proteomic characterization of human mes-
stemcells.nih.gov. (Accessed 21 April 2017). enchymal stem cells reveals source specific cellular markers, Sci
6. Thomson JA, Itskovitz-Eldor J, Shapiro SS, et al: Embry- Rep 6:21507, 2016.
onic stem cell lines derived from human blastocysts, Science 26. Phinney DG: A SAGE view of mesenchymal stem cells, Int J
282(5391):1145–1147, 1998. Stem Cells 2(1):1–10, 2009.
7. Ben-David U, Benvenisty N: The tumorigenicity of human 27. Kolf CM, Cho E, Tuan RS: Mesenchymal stromal cells. Biology
embryonic and induced pluripotent stem cells, Nat Rev Cancer of adult mesenchymal stem cells: regulation of niche, self-renewal
11:268–277, 2011. and differentiation, Arthritis Res Ther 9(1):204, 2007.
8. Selvaraj V, Wildt DE, Pukazhenthi BS: Induced pluripotent 28. Siniscalco D, Giordano C, Galderisi U, et al: Long-lasting effects
stem cells for conserving endangered species? Nat Methods of human mesenchymal stem cell systemic administration on
8(10):805–807, 2011. pain-like behaviors, cellular, and biomolecular modifications in
9. Ben-Nun IF, Montague SC, Houck ML, et al: Induced plu- neuropathic mice, Front Integr Neurosci 5:79, 2011.
ripotent stem cells from highly endangered species, Nat Methods 29. Guo W, Imai S, Dubner R, et al: Multipotent stromal cells
8(10):829–831, 2011. for arthritic joint pain therapy and beyond, Pain Manag
10. Prescott HMA, Manning C, Gardner A, et al: Giant Panda 4(2):153–162, 2014.
(Ailuropoda melanoleuca) buccal mucosa tissue as a source 30. Kol A, Wood JA, Holt DDC, et al: Multiple intravenous injec-
of multipotent progenitor cells, PLoS ONE 10(9):e0138840, tions of allogeneic equine mesenchymal stem cells do not induce
2015. a systemic inflammatory response but do alter lymphocyte subsets
11. Saragusty J, Diecke S, Drukker M, et al: Rewinding the process in healthy horses, Stem Cell Res Ther 6(1):73, 2015.
of mammalian extinction, Zoo Biol 35(4):280–292, 2016. 31. Owens SD, Kol A, Walker NJ, et al: Allogeneic mesenchymal
12. Caplan AI: Mesenchymal stem cells, J Orthop Res 9(5):641–650, stem cell treatment induces specific alloantibodies in horses, Stem
1991. Cells Int, 2016.
13. Dominici M, Le Blanc K, Mueller I, et al: Minimal criteria 32. Harman R, Carlson K, Gaynor J, et al: A prospective, ran-
for defining multipotent mesenchymal stromal cells. The domized, masked, and placebo-controlled efficacy study of
International Society for Cellular Therapy position statement, intraarticular allogeneic adipose stem cells for the treatment of
Cytotherapy 8(4):315–317, 2006. osteoarthritis in dogs, Front Vet Sci 3:81, 2016.
14. Lee TC, Lee TH, Huang YH, et al: Comparison of surface 33. Caplan AI: All MSCs are pericytes? Cell Stem Cell 3(2):229–230,
markers between human and rabbit mesenchymal stem cells, 2008.
PLoS ONE 9(11):e111390, 2014. 34. de Souza LE, Malta TE, Kashima Haddad S, et al: Mesenchymal
15. Sago K, Tamahara S, Tomihari M, et al: In vitro determination stem cells and pericytes: to what extent are they related? Stem
of canine celiac adipose tissue-derived stromal cells into neuronal Cells Dev 25(24):1843–1852, 2016.
cells, J Vet Med Sci 70(4):353–357, 2008. 35. Friedenstein AJ, Chailakhyan RK, Gerasimov UV: Bone marrow
16. Bourin P, Bunnell BA, Casteilla L, et al: Stromal cells from osteogenic stem cells: in vitro cultivation and transplantation in
the adipose tissue-derived stromal vascular fraction and culture diffusion chambers, Cell Tissue Kinet 20(3):263–272, 1987.
expanded adipose tissue-derived stromal/stem cells: a joint state- 36. Taylor SE, Clegg PD: Collection and propogation methods for
ment of the International Federation of Adipose Therapeutics mesenchymal stromal cells, Vet Clin North Am Equine Pract
(IFATS) and Science and the International Society for Cellular 27(2):263–274, 2011.
Therapy (ISCT), Cytotherapy 15(6):641–648, 2013. 37. Sampaio RV, Chiaratti MR, Santos DC, et al: Generation of
17. Caplan AI, Correa D: The MSC: an injury drugstore, Cell Stem bovine (Bos indicus) and buffalo (Bubalus bubalis) adipose tissue
Cell 9(1):11–15, 2011. derived stem cells: isolation, characterization, and multipotenti-
18. Amarpal A, Dhama K, Chakraborty S, et al: Stem cells and their ality, Genet Mol Res 14(1):53–62, 2015.
clinical/therapeutic applications in biomedical and veterinary 38. Johnson SP, Catania JM, Harman RJ, et al: Adipose-derived
science – the perspectives, Res Opin Anim Vet Sci 3(9):261–279, stem cell collection and characterization in bottlenose dol-
2013. phins (Tursiops truncatus), Stem Cells Dev 21(16):2949–2957,
19. Caplan AI: Review: mesenchymal stem cells: cell based recon- 2012.
structive therapy in orthopedics, Tissue Eng 11(7–8):1198–1211, 39. Chong PP, Selvaratnam L, Abbas AA, et al: Human peripheral
2005. blood derived mesenchymal stem cells demonstrate similar
20. Spees JL, Lee RH, Gregory CA: Mechanisms of mesenchy- characteristics and chondrogenic differentiation potention to
mal stem/stromal cell function, Stem Cell Res Ther 7(1):125, bone marrow derived mesenchymal stem cells, J Orthop Res
2016. 30(4):634–642, 2012.
21. Prockop DJ, Kota DJ, Bazhanov N, et al: Evolving paradigms 40. Spaas JH, De Schauwer C, Cornillie P, et al: Culture and charac-
for repair of tissues by adult stem/progenitor cells (MSCs), J Cell terisation of equine peripheral blood mesenchymal stromal cells,
Mol Med 14(9):2190–2199, 2010. Vet J 195(1):107–113, 2013.
22. Caplan AI, Dennis JE: Mesenchymal stem cells as trophic media- 41. Karp JM, Leng Teo GS: Mesenchymal stem cell homing: the
tors, J Cell Biochem 98(5):1076–1084, 2006. devil is in the details, Cell Stem Cell 4(3):206–216, 2009.
144 SE C T I O N 5 Therapeutics
42. Sohni A, Verfaillie CM: Mesenchymal stem cells migration 53. Alves AG, Stewart AA, Dudhia J, et al: Cell-based therapies for
homing and tracking, Stem Cells Int 2013:130763, 2013. tendon and ligament injuries, Vet Clin North Am Equine Pract
43. Saito T, Kuang JQ, Lin CC, et al: Transcoronary implantation 27(2):315–333, 2011.
of bone marrow stromal cells ameliorates cardiac function after 54. Miller PI, Clegg PD, Stewart MC: Stem cell-based therapies for
myocardial infarction, J Thorac Cardiovasc Surg 126(1):114–123, bone repair, Vet Clin North Am Equine Pract 27(2):299–314,
2003. 2011.
44. François S, Bensidhoum M, Mouiseddine M, et al: Local irradia- 55. Rich RF: Single center study of 83 horses with suspensory
tion not only induces homing of human mesenchymal stem cells injuries treated with adipose-derived stem and regenerative cells,
at exposed sites but promotes their widespread engraftment to Stem Cell Discovery 4:44–53, 2014.
multiple organs: a study of their quantitative distribution after 56. De Schauwer C, Van de Walle GR, Van SA, et al: Mesenchymal
irradiation damage, Stem Cells 24(4):1020–1029, 2006. stem cell therapy in horses: useful beyond orthopedic injuries?
45. Harman R, Ross R, Schaffer J: Adipose-derived stem cells given Vet Q 33(4):234–241, 2013.
intravenously improves epistaxis and objective bronchoscopy 57. Harman RJ: Stem cell therapy in veterinary dermatology, Vet
scoring in horses with exercise-induced pulmonary hemorrhage, Dermatol 24(1):90–96, 2013.
November 17–20, 2016. North American Veterinary Regen- 58. Arzi B, Mills-Ko E, Verstraete FJ, et al: Therapeutic efficacy of
erative Medicine Association Meeting. Channel Islands, Florida, fresh, autologous mesenchymal stem cells for severe refractory
USA. gingivostomatitis in cats, Stem Cells Transl Med 5(1):75–86,
46. Kang SK, Shin IS, Ko MS, et al: Journey of mesenchymal stem 2016.
cells for homing: strategies to enhance efficacy and safety of stem 59. Hoffman AW, Dow SW: Concise review: stem cell trials using
cell therapy, Stem Cells Int 2012:342968, 2012. companion animal disease models, Stem Cells 34(7):1709–1729,
47. Black LL, Gaynor J, Gahring D, et al: Effect of adipose-derived 2016.
mesenchymal stem and regenerative cells on lameness in dogs 60. Puliafico SB, Penn MS, Silver KH: Stem cell therapy for heart
with chronic osteoarthritis of the coxofemoral joints: a random- disease, J Gen Intern Med 28(10):1353–1363, 2013.
ized, double-blinded, multicenter, controlled trial, Vet Ther 61. Tang YH, Ma YY, Zhang ZJ, et al: Opportunities and challenges:
8(4):272–284, 2007. stem cell-based therapy for the treatment of ischemic stroke, CNS
48. Vilar JM, Morales M, Santana A, et al: Controlled, blinded Neurosci Ther 21(4):337–347, 2015.
force platform analysis of the effect of intraarticular injection of 62. Peired AJ, Sisti A, Romagnani P: Mesenchymal stem cell-based
autologous adipose-derived mesenchymal stem cells associated to therapy for kidney disease: a review of clinical evidence, Stem
PRGF-Endoret in osteoarthritic dogs, BMC Vet Res 9(131):2013. Cells Int 2016:4798639, 2016.
49. Vilar JM, Cuervo B, Rubio M, et al: Effect of intraarticular 63. Sivan PP, Syed S, Mok PL, et al: Stem cell therapy for treatment
inoculation of mesenchymal stem cells in dogs with hip osteo- of ocular disorders, Stem Cells Int 2016:8304879, 2016.
arthritis by means of objective force platform gait analysis: 64. El-Badawy A, El-Badri N: Clinical efficacy of stem cell therapy
concordance with numeric subjective scoring scales, BMC Vet for diabetes mellitus: a meta analysis, PLoS ONE 11(4),
Res 12:223, 2016. 2016.
50. Mohoric L, Zorko B, Ceh K, et al: Blinded placebo study of 65. Berardis S, Sattwika PD, Najimi M, et al: Use of mesenchymal
bilateral osteoarthritis treatment using adipose derived mesen- stem cells to treat liver fibrosis: current situation and future
chymal stem cells, Slov Vet Res 53(3):167–174, 2016. prospects, World J Gastroenterol 21(3):742–758, 2015.
51. Nixon A, Dahlgren LA, Haupt JL, et al: Effect of Adipose- 66. Broeckx S, Borena BM, Zimmerman M, et al: Intravenous
derived nucleated cell fractions on tendon repair in horses with application of allogenic peripheral blood-derived mesenchymal
collagenase-induced tendinitis, Am J Vet Res 69:928–937, 2008. stem cells: a safety assessment in 291 equine recipients, Curr Stem
52. Frisbie DD, Steward MC: Cell-based therapies for equine joint Cell Res Ther 9(6):452–457, 2014.
disease, Vet Clin North Am Equine Pract 27(2):335–349, 2011.
25
Compounding Pharmacies
KATHRYN C. GAMBLE
C
ommercial drug manufacturing commences with an includes the various basic manipulations by which a new
active pharmaceutical ingredient (API), also known formulation of a currently FDA-approved drug can be
as a bulk drug. In this terminology, it does not created.5,7,12,13 Most simply, it can be an in-clinic processing
mean a measure of volume but that it is the base ingredi- such as grinding a tablet into a powder to facilitate admin-
ent.1 Inert additives (such as fillers, excipients, or colorants) istration in a patient’s food (change of formulation) or
or active agents that affect drug behavior may be added diluting a parenteral product with sterile saline to facilitate
during production of a marketed pharmaceutical.1 In the more accurate measurement for a smaller patient (change
United States, in its final formulation, a pioneer drug will of concentration).13 To be clear, any action performed
be approved by the Food and Drug Administration (FDA), that is not specifically on the label direction is considered
after legally required time-consuming and costly evaluations compounding. This includes reduction in the quantity of
demonstrate that it complies with its intended safety and reconstitution diluent for concentrated end use or simply
efficacy parameters.1,2 mixing two drugs together in one syringe.7,14
Inherent in these factors are potency (actual concentra- Professional compounding is available for more difficult
tion of active ingredient per unit of drug formulation),2–7 manipulations such as creating sterile parenteral anesthetic
accuracy (difference between actual and intended con- solutions and resurrection of drugs absent from the market
centration, and specifically ±10% of labeled amount),3 using APIs unavailable in the nonpharmacist role; or chang-
precision (variation around the mean of concentrations),3 ing a commercial oral formulation to that of a transdermal
quality (absence of harmful contaminants or ingredients gel.5 Although these CPs may be evaluated independent of
other than labeled),2 purity (inclusion of only intended federal oversight, they remain unapproved by FDA regula-
product),5 integrity (retention of potency until beyond tions and thus are not ensured to create their intended effect
use date),2 and lack of contamination (inclusion of actual when used in the course of patient care.3,6,7 In addition,
infectious agents).4,5 In this process, both medical and it is important to understand that use of CPs, even in
veterinary pharmaceuticals are labeled with specifically the labeled species of the original formulation, and the
evaluated doses, routes of administration, indications, and, use of FDA-approved drugs for any indication or species,
for veterinary medicine, particular species applicable to the or by any route that is not specifically labeled, constitute
testing completed, making these expressly legal uses. After extralabel drug use (ELDU). In the United States, as the
such a drug has reached the completion of its patented life prescriber, the veterinarian has the privilege of selecting the
span, generic drugs may become available that are developed pharmaceutical best suited for his or her patients’ needs,
by these same manufacturing processes. These products also whether this be a medical or veterinary pharmaceutical, or
are FDA approved and required to be bioequivalent copies FDA-approved or compounded drug. As such, it is critical
of the original product.1,2,8 Due to market interest and that the veterinary clinician fully understand the incumbent
commercial marketing pressures, subsequently identified liability as the prescriber, particularly with CPs.2,6
adverse drug events (ADEs), or product innovations, these
pioneer drugs and their generic equivalents may become
temporarily or permanently unavailable to the end user.9 Pharmaceutical Legislative History and
It is into and within all of these means and ways that Current Perspectives1,5–8,10,14
compounded pharmaceuticals (CPs) can be found both
integrated, yet separate. Although the following discussion is focused on the per-
In the subsequent discussion, CPs are drug formula- spective within the United States, it should be considered
tions developed from either API or the finished commercial that the basic issues arising from drug manufacturing
or generic formulations, by any change from the labeled and compounding will be present throughout worldwide
procedures or directions.1,5–7,10 Traditional compounding pharmaceutical regulations where they exist.7 In 1938,
145
146 SE C T I O N 5 Therapeutics
due to rising concern over contaminated pharmaceuticals opportunities were not used.1 As such, this publication was
and poor production standards, legislative action resulted rescinded in its entirety in May 2015.1,14 Although a victory
in the Federal Food, Drug, and Cosmetic Act (FFDCA, in many ways, the compounding process now remains
21 U.S.C. § 360) and created the FDA as a regulatory body without federal regulation, guidance, or acceptance, leaving
for drug manufacturing. These legislations largely ignored front-line clinicians in a gray zone of prescription liability.
veterinary medicine in specific mention until its amendment As of now, only a Guidance for Industry (GFI #230) docu-
in 1968.2,11 Implemented and unaltered for decades, this ment has been drafted and remains unfinalized.7,11
initial legislation did not encompass compounding either as
a procedure or end product, although it was well known that Compounding—What It Is and Is Not
these activities were occurring for patient care. However, as
veterinary medicine sophistication increased, practitioners Compounding is increasing, at quite startling rates, within
used more medical pharmaceuticals and administered all medical fields. Essentially any pharmacy can perform
veterinary drugs to species other than those labeled in the compounding, although professional compounders focus
commercial manufacturing process. Therefore to address their procedures in these endeavors. Veterinary compound-
this regulatory deficiency, the Animal Medicinal Drug Use ing can occur at either medical or veterinary pharmacies,
Clarification Act (AMDUCA; 21 C.F.R. § 530.13) was although veterinary sources are considered to have deeper
passed in 1994, which provided veterinarians legal support understanding of the variable patient species base.6,7
to select, administer, and dispense medications in an ELDU Ideally, the FDA intended that drug compounding
fashion.12 It included acknowledgment of compounding should not even resemble manufacturing. It is expected
when no FDA-approved pharmaceutical was available.7 to occur for an individual patient from a specific prescrip-
Although no historic reference existed, it provided that with tion generated by a veterinarian within a valid professional
adherence to all relevant state laws and with professional relationship with both a client and pharmacist.5 Although
direction from veterinarian to pharmacist, compounding formulating new drugs from FDA-approved manufactured
was permissible; however, specific exclusion was made limit- products, compounding is not to be confused with the
ing compounding from APIs. This enhanced permission creation of new drugs. As such, the safest means to avoid
and the burgeoning standards of veterinary care and client legal contention is use of an FDA-approved commercial or
expectations continued market pressure on compounding generic product for compounding.2 However, this approach
pharmacies to provide more individualized products and is not always appropriate, as these formulations may not
formulations. lend themselves to change dramatically between routes
Understandably, the FDA and commercial manufactur- of administration in a manner safe for the patient.6,7,11
ers as represented by the Animal Health Institute (AHI) Additives in the finalized products may be undesired, such
were concerned about substandard activities in this federally as beef-based flavorings for food-intolerant individuals or
unregulated field, especially when quite public, traceable veterinary-specific additives of concern (e.g., xylitol) that
ADEs, or failures of treatment occurred. Admittedly, one cannot be separated through a reliable procedure from the
must consider that financial competition had some role commercial preparations.11,16 Furthermore, in situations of
in this opinion and resultant regulatory pressure. In 2003 discontinued commercial availability, the finished product
FDA promulgated its Compliance Policy Guide (CPG, actually no longer exists.9 In these situations, compounders
specifically 608.400) as replacement for its 1992 guide to obtain APIs as their base to begin the process. Although
its employees and actions and specifically sought to assess not expressly forbidden, this practice is officially within an
and address potential compounding violations. In particu- FDA-unapproved area.10
lar, compounding from APIs and question of appropriate Immediately, this approach calls attention to itself
volume of reserves of CPs based on sales rather than for because it can be considered manufacturing to take a bulk
specific patient need were at issue. Unfortunately, this guide product and formulate a marketed end product. However,
was taken in some arenas as actual legislated authority. in compounding, APIs can be the desired base to eliminate
Furthermore, it had many areas of vague definition and concerns of purity, need for extraction of additives, and
contradiction in purpose, which actually confounded rather resolve lack of availability. As occurs in manufacturing,
than clarified the compounding issue. It particularly isolated these APIs should be obtained from an FDA-registered
species-care groups, such as exotic animal practitioners, and source for optimal assurance of quality and for reduced
prevented their voice in patient care needs. legal liability.2
Since 2000, repeated testimony and veterinary industry Under these circumstances, regulatory concern should ebb
lobbying from multiple disciplines encouraged congres- because it is not a bypass route to the drug-manufacturing
sional conclusion that the CPG was unfounded in its and FDA approval process but rather provision of a needed
current form.14 Inconsistent enforcement was documented, pharmaceutical. Furthermore, compounding from APIs has
especially directed at compounding sources1; excessive and played a role in regulatory defined minor species—essentially
unnecessary product labeling was a concern15; and where all those in zoo and wildlife veterinary care—where market
documentation through established routes, such as ADEs, share will never justify the pursuit of commercially avail-
could be made to accumulate scientific evidence, these able finished products through FDA-approved procedures.
CHAPTER 25 Compounding Pharmacies 147
Although closer to manufacturing than traditional com- and may enhance risk to the patient or administrator,
pounding, it remains a necessary consideration that should particularly noteworthy in transdermal preparations.6,7,11
not raise regulatory hackles, although protest of market Of greater concern is the expectation by the end user that
diversion has been made from commercial players. However, CPs are evaluated consistently by some party to ensure the
of particular caution, compounding API use as a blatant label information is accurate. In contrast, repeated random
means to undercut commercial products by mimicry is assessment by federal and state regulatory bodies and inde-
considered piracy. Low-cost sourcing of APIs to provide pendent scientific publications have demonstrated that not
a final product below cost of the commercial version is only between but also within batch irregularity exists.3,7,9
considered ethically inappropriate and may produce ques- Overall concerns for variability in potency have been dem-
tionable efficacy if API quality is compromised for reduced onstrated routinely, and in rare instances, actual absence or
price.8,14 Ongoing and often contentious discussions multiple-fold presence of API has been confirmed.1,4,5
between AHI and compounding pharmacists have occurred Furthermore, without the rigor of testing, it is unknown
over consideration of larger volumes of drugs prepared from what the best use date (BUD) may be for a given CP, unlike
bulk sources that are held in reserve. By federal review, it the stability testing required of commercial and generic
was concluded that available compounded inventory could products. Difference between large-volume manufacturing
be necessary for rapid response of the compounding source to small-volume compounding steps may produce differ-
to its clients, as would be expected for good commercial ent results in the same product formulation. It has been
manufacturing practice. This approach should be tailored published that CPs cannot be reliably assigned a BUD
to routine market need rather than mass-volume sales. If past 14 days, unless specifically evaluated.2,4,6,9,12,16,17,20–22 In
only these deciding factors faced prescribing veterinarians, particular, USP standards recommend that nonsterilized,
it would be a minimal minefield from which to proceed. water-based oral formulations be stored at controlled cold
But what quality standards exist where no standards are temperatures (2°C–8°C) to even permit the assignment of
actually mandated? this BUD, and consideration that pH changes or exposure
Initially, the end-use concern of poor product sourcing to UV light unexpectedly can affect this recommenda-
should be eliminated by engagement of pharmacies that tion.3,6,7,12,17,21 In some situations, commercial additives
operate under API sourcing guidelines provided to manu- are active and necessary for appropriate pharmacodynamic
facturing facilities. In addition, assurance of expected end expectations; when these agents are proprietary, and there-
result can be enhanced by prescribing from compounders fore unavailable to the compounder, the resultant CP is
using industry standards of the US Pharmacopeial Conven- not as effective as the commercial product or even the
tion (USP),7,17 which is a not-for-profit scientific organiza- originating API. Often, it has required treatment failure in
tion that is referenced by FDA regulation.4,5 Initiated in repeated situations to generate sufficient interest to perform
1820, USP has gathered methodologies and details accepted scientific investigation to validate these discrepancies. An
as good practice basis and is accepted in more than 140 outstanding example of this concern is compounded
countries worldwide. It details not only production but itraconazole, which lacks the essential cyclodextrin moiety.
also chemical properties and cautions, such that adher- The resultant diminished serum concentrations in clinical
ence to the guide should produce a better end product.1,5 patients, and repeated failure of efficacy, have resulted in
This document has been updated to include a dedicated the recommendation that compounding not be used in this
chapter to veterinary compounding.6 Although the USP is product.6,22 Although these studies have been quite limited
not uniformly required for use in all states and territories in exotic patient situations, it is within the wide variety of
within the United States,1,5 compounding regulation for these species that one easily can extrapolate that similar
pharmacies does exist at a state level. However, it is widely concerns can exist in any compounding process. In each
variable in application, so prescribers must be aware of their of these situations, rather than enhance patient care, CPs
state’s regulations.7,18 These vague boundaries of veterinary compromise clinical outcome and produce ADE potential
compounding are complicated further when facilities ship for which the veterinary prescriber is liable.6
across state lines. An additional measure of assurance is
that compounding does have self-policing via various Appropriate Compounding Situations
professional organizations, including the Pharmacy Com-
pounding Accreditation Board (PCAB) established from Federal recognition exists that compounding is necessary
industry sources to create compounding regulations; and the in veterinary medicine. Use of CPs must ensure that harm
International Academy of Compounding Pharmacists and and therapeutic failure do not occur to the patient and that
American College of Veterinary Pharmacists that provide no violative residues are produced in food source animals.6
training, continuing education, and certification.4,5,10 However, as considered and recommended by both Ameri-
Yet, as non–FDA-approved products, CPs have no guar- can Veterinary Medical Association and American Associa-
antee of safety or efficacy, even when produced with care.2,10 tion of Zoo Veterinarians documentation,8 compounding
Sterile formulations for parenteral use have been made that is appropriate in many situations but must include a valid
presented issues of fatal contamination.19 Novel formula- veterinary-client-patient relationship and have expected
tions have been created that are scientifically unproven prevention of animal death and suffering and, wherever
148 SE C T I O N 5 Therapeutics
possible, be patient specific by prescription.1,10 In particular, in 2013, consideration of compounding situations for this
compounding is condoned when it is: setting must be enumerated.14
1. Focused on those products that have been determined As mentioned, accuracy of dosing for both large and
safe and effective in target species in their compounded small patients is facilitated by CPs. Immediate availability of
form, and the risks of lack of efficacy or expectation of antiinfective products in their appropriate form or at-hand
ADEs are outweighed by potential patient benefit; availability of appropriate anesthetics in the face of escape
2. An adjustment of formulation to accommodate indi- or urgent need precludes the ability of this discipline to
viduals where no other available formulation will be use prescriptions of individual and volume-limited CPs.
successful; and However, each zoo and wildlife veterinarian is charged to
3. Possible to monitor the effects of CPs through indirect exercise this part of their duties with the understanding that
measures of physiologic function, or actual therapeutic each of our actions has the potential to affect the entire
drug monitoring (TDM).9 discipline if sound judgment is abandoned in the face of
Specific indications from these considerations include: convenience. Counsel in the use of CPs, and thorough
1. Lack of appropriate drug concentration that requires documentation, should be made in the practice setting.
adjustment to facilitate either more precise dose mea- Veterinarians should be critical of their sources of CPs.
surement (smaller patients) or administration of more This caution is not intended in a negative or punitive
appropriately sized dose formulations (larger patients); manner but rather encourages an educated and construc-
2. Lack of appropriate formulation in what is available tive communication. It should be ensured what state and
commercially; industry oversight that is available will be exercised; mea-
3. Accessing essential treatment from drugs that are tem- sures of good practices are engaged by personally selected
porarily or permanently removed from market access for compounders; and a relationship should be maintained to
reasons other than ADEs in veterinary patients; and identify concerns or ADEs more quickly by either prescriber
4. Accessing essential drugs for minor species, as consistent or compounder. Appropriateness of the situation and the
with the definition established in 1994 by the Minor Use ethical impact of the CP choice is tantamount to the suc-
and Minor Species (MUMS) Animal Health Act.1 cessful prescription. In this consideration the factor of larger
Drug compounding does have inappropriate applica- volumes maintained for office dispensing should consider
tions. Veterinarians are advised to avoid this approach that BUD has not been determined for many CPs, such that
when: any product provided a longer than 14-day BUD may need
1. Commercial or generic products in their marketed form to be considered for more rapid disposal and replacement.
are satisfactory and appropriate for the intended use, Published literature on compounded formulations, even in
aside from minor in-office manipulation; domestic species, should be reviewed regularly to ensure
2. Food-producing animals are involved, including all that known CP issues are incorporated into or avoided in
cattle, swine, domesticated poultry, sheep, goats, deer, patient care.3,12,16,20,22 In addition, initiation of such analyses
rabbits, and nonornamental fish, even when these species is needed sorely, and it should span many species and indi-
are pet breeds or managed in the captive setting. These cations.1 Participation in legislative updates (https://www
species are restricted due to concerns of food source .avma.org/kb/resources/reference/pages/compounding
contamination from either intentional or inadvertent .aspx) by the practitioner needing CPs is critical to inform
production streams.1 Exceptions have been permitted in future regulatory processes.
depopulation, euthanasia, and specific antidotes that as
such will not enter the food stream1; Conclusion
3. Production is based solely on the ability to reduce cost
of the treatment by the process of compounding2; and In conclusion, the prescriber must be engaged in the process
4. Minimal relationship is available with the compound- of prescription—whether this be for a compounded or
ing source such that understanding of their practices is FDA-approved drug. The responsibility to select the optimal
unknown. drug, appropriate formulation, and advocated administra-
tion route to their best ability requires that veterinarians
understand the risks of any prescription administered.
Compounding Relevance and Furthermore, when considering compounding, it should be
Considerations for the Zoo and questioned whether compounding is appropriate—or even
Wildlife Specialty necessary.11 In addition, diligence in reporting prescrip-
tions and treatment success or failure are important within
It scarcely needs mention to this audience that compound- animal records, despite the unclear regulations existing on
ing is critically essential to patient care. Available drugs CP labeling. Regardless of the originating pharmaceutical,
minimally are labeled for the vast array of species and in the face of treatment failure or unexpected outcome, it
medical concerns of this discipline. Yet for completeness, should be considered that the prescription was the source.
and because only 5% of the total compounding effort in As such, antemortem treatment can be adjusted or post-
the United States was directed to zoo and wildlife species mortem evaluation can confirm the issue. Thorough ADE
CHAPTER 25 Compounding Pharmacies 149
reporting within 15 days is recommended to not only the 10. American Veterinary Medical Association: Veterinary Compound-
pharmacy source but also the FDA (www.fda.org; Form ing. Available at: https://ebusiness.avma.org/ProductCatalog/
FDA 1932a), even for compounded drugs that currently product.aspx?ID=155. (Accessed 3 June 2017).
11. Eichstadt LR: Compounding transdermal medications for feline
do not have this legal requirement.1,10 By doing so, one not
patients, Int J Pharm Comp 20:271–274, 2016.
only provides excellence in individual practice care, but also 12. Petritz OA, Guzman DS-M, Wiebe VJ, et al: Stability of three
precludes additional harm to similar animals or species in commonly compounded extemporaneous enrofloxacin suspen-
global zoo and wildlife veterinary care. sions for oral administration to exotic animals, J Am Vet Med
Assoc 243:85–90, 2013.
References 13. Frank H: Compounding in the exotic practice, J Exot Pet Med.
15:116–121, 2006.
1. Drug compounding for animals: FDA could improve oversight 14. Brakke Consulting: Veterinary drug compounding survey, 2013.
with better information and guidance; GAO-15-671. Available Available at: http://www.brakkeconsulting.com//veterinary-drug
at: http://www.gao.gov/assets/680/672748.pdf. (Accessed 3 June -compounding.aspx. (Accessed 3 June 2017).
2017). 15. FDA Proposed Guidance Document, Compounding animal drugs
2. Stanley SD: Legal and ethical veterinary compounding. Available from bulk drug substances. Available at: http://c.ymcdn.com/
at: http://docplayer.net/12674614-Legal-and-ethical-veterinary sites/www.iacprx.org/resource/resmgr/FDA/May_20,_2015_
-compounding-scott-d-stanley-ph-d-professor-university-of FDA_Draft_Vet_C.pdf. (Accessed 3 June 2017).
-california-davis-school-of-veterinary-medicine.html. (Accessed 16. Baine K, Jones MP, Cox S, et al: Pharmacokinetics of compounded
3 June 2017). intravenous and oral gabapentin in Hispaniolan Amazon parrots
3. Laporte CM, Cruz-Espindola C, Thungrat K, et al: Quality (Amazona ventralis), J Avian Med Surg 29:165–173, 2015.
assessment of fluconazole capsules and oral suspension com- 17. Davidson G: USP compounding standards: yes, they do apply
pounded by pharmacies located in the United States, Am J Vet to veterinary compounding, in Proceedings. 34th Annu Meet Soc
Res 78:421–431, 2017. Vet Hosp Pharmacists.
4. Pollock C: Compounding in clinical avian practice, J Av Med 18. American Veterinary Medical Association State summary report.
Surg 30:196–202, 2016. Available at: https://www.avma.org/Advocacy/StateAndLocal/
5. Glassgold JM: Congressional Research Service, Compounded Pages/compoundinglaws.aspx. (Accessed 3 June 2017).
drugs, 2013. Available at: https://fas.org/sgp/crs/misc/R43082 19. Vasquez AM, Lake J, Ngai S, et al: Notes from the field:
.pdf. (Accessed 3 June 2017). fungal bloodstream infections associated with a compounded
6. Papich M: Drug compounding for veterinary patients, Am Assoc intravenous medication at an outpatient oncology clinic – New
Pharm Scient J 7:E281–E287, 2005. York City, 2016, MMWR Morb Mortal Wkly Rep 65:1274–1275,
7. Davidson G: Veterinary compounding: regulation, chal- 2016.
lenges, and resources, Pharmaceutics. 9:5, 2017. doi:10.3390/ 20. Papich MG, Davidson GS, Fortier LA: Doxycycline concentra-
pharmaceutics9010005 tion over time after storage in a compounded veterinary prepara-
8. White-Shim L: Compounding – what’s happening and what’s the tion, J Am Vet Med Assoc 242:1674–1678, 2013.
AVMA doing? (7 July 2014). Available at: https://www.avma.org/ 21. Davis JL, Kirk LM, Davidson GS, et al: Effects of compounding
Search/results.aspx?k=White-Shim%20L.%20%20Compound- and storage conditions on stability of pergolide mesylate, J Am
ing. (Accessed 3 June 2017). Vet Med Assoc 234:385–389, 2009.
9. Scott-Moncrief JCR, Moore GE, Coe J, et al: Characteristics of 22. Smith JA, Papich MG, Russell G, et al: Effects of compounding
commercially manufactured and compounded protamine zinc on pharmacokinetics of itraconazole in black-footed penguins
insulin, J Am Vet Med Assoc 240:600–605, 2012. (Spheniscus demersus), J Zoo Wildl Med 41:487–495, 2010.
SECTION 6
150
26
Sustained-Release and Long-Acting
Opioid Formulations of Interest in
Zoological Medicine
JESSICA A. EMERSON AND DAVID SANCHEZ-MIGALLON GUZMAN
151
152 SE C T I O N 6 Anesthesia and Analgesia
TABLE
26.1 Sustained-Release Opioid Drugs of Interest in Zoological Medicine
Fentanyl Transdermal 25 µg/h; 50 µg/h PK and PD13 (post- Once, in place for 72 h
patch operative left lung
allograft transplant)
CHAPTER 26 Sustained-Release and Long-Acting Opioid Formulations of Interest in Zoological Medicine 155
Domestic swine Skin behind ear, canvas N = 8, 20–30 kg pigs, no sedation noted, large variations in serum
(growing) sutured over patch for fentanyl concentrations, no difference in absorption associated with
protection; placed after anesthesia
30 min of isoflurane
anesthesia or no
anesthesia
Domestic swine Dorsal interscapular region N = 3 at 25 µg/h, N = 2 at 50 µg/h; pigs treated with 25 µg/h had
higher pain scores and lower serum concentrations (0.1–0.3 ng/mL);
pigs treated with 50 µg/h had the lowest pain scores and serum
concentration >0.5 ng/mL
Continued
156 SE C T I O N 6 Anesthesia and Analgesia
TABLE
26.1 Sustained-Release Opioid Drugs of Interest in Zoological Medicine—cont’d
Fentanyl Transdermal 2 µg/kg IV; 2 µg/kg/h PK and PD17 Once IV and once
patch transdermal (varying transdermally, in place for
combinations of patch sizes 72 h
used based on weight of
animal)
Fentanyl Transdermal 2 µg/kg/h (varying PD14 Once; in place for 72 h
patch combinations of patch sizes
used based on weight of
animal)
Fentanyl Transdermal 2.5 µg/kg IV; 2.05 µg/kg/h PK15 Once; patch in place for 72 h
patch and transdermal (varying combi-
IV nations of patch sizes used
based on weight of animal)
Fentanyl Transdermal 50 µg/h PK18 Once IV (2.5 µg/kg); Once
patch transdermally, in place for
72 h
Buprenorphine Transdermal 35 µg/h PK and PD56 (preliminary, Once, in place for 72 h
thermal threshold)
Domestic sheep Lateral antebrachium N = 15, unilateral tibial osteotomy performed 12 h after placement,
superior analgesia to 0.01 mg/kg buprenorphine q6h IM
Domestic sheep Lateral antebrachium N = 6 IV, N = 15 transdermal; maximum plasma concentration reached
at median of 12 h and a median level of 1.3 ng/mL, plasma
concentrations >0.5 ng/mL for 40 h after patch placement
Domestic goats Right lateral neck, covered N = 6; variable plasma concentrations, time to peak concentration
with gauze and elastic tape ranged from 8 to 18 h, from 4 to 36 h the plasma concentration
around the neck to secure remained >2 ng/mL, but then declined so a steady state was never
achieved
Domestic chickens Left iliopsoas muscle N = 10; marked individual variability, all chickens had levels within target
following feather plucking range (0.2–1.2 ng/mL) through 72 h, peak plasma concentrations of
2.86 ± 2.58 ng/mL at 14.9 ± 8.2 h after placement
Helmeted guineafowl Interscapular skin N = 21, no adverse effects or changes in behavior, Mean peak plasma
(Numida meleagris) concentration 228.8 ng/mL at 4 h, plasma concentrations >0.6 ng/mL
for at least 7 days
Ball pythons (Python Dorsal mid-body, attached N = 2, plasma concentrations >1 ng/mL within 4 h
regius) with 4 staples
Ball pythons (Python Epaxial musculature lateral to N = 16, no evidence of thermal antinociception at either dose, evidence
regius) spine of respiratory depression, plasma concentration >1 ng/mL within 6 h
Continued
158 SE C T I O N 6 Anesthesia and Analgesia
TABLE
26.1 Sustained-Release Opioid Drugs of Interest in Zoological Medicine—cont’d
Buprenorphine Transdermal 70 µg/h PD53 (post-operative pain) Once, in place for 86 h (applied
48 h preoperatively)
Buprenorphine SC Injection 0.12 mg/kg PD40 Once, immediately
SR-LAB preoperatively
Buprenorphine Transdermal 30 µg/h (1 each of 20 µg/h PK49 Once, in place for 72 h
and 10 µg/h)
FDA, US Food and Drug Administration; IM, intramuscularly; IV, intravenous; OHE, ovariohysterectomy; PD, pharmacodynamics study; PK, pharmacokinetic
study; SC, subcutaneous; SR, sustained-release.
CHAPTER 26 Sustained-Release and Long-Acting Opioid Formulations of Interest in Zoological Medicine 159
TABLE
26.2 Anecdotal Reports of Sustained-Release Opioid Formulations in Zoological Medicine
SR, Sustained-release.
Buprenorphine transdermal patches have been evalu- Forty-eight hours after both IM and SC administration,
ated in dogs,53–55 pigs,49 and cats.56 Overall, target plasma mean plasma concentrations remained greater than target
concentrations have been reached and resulted in pain concentrations (>1 ng/mL). SC hematomas in three of
control in dogs undergoing ovariohysterectomy,53 as well the birds that received buprenorphine SR-LAB SC were
as antinociceptive effects using thermal thresholds.55 In attributed to traumatic administration but resolved without
cats, there was no antinociceptive effect noted using the intervention. A follow-up study concluded that depending
thermal threshold model.56 Interestingly, undetectable on the severity and type of pain, adjunctive therapy, and
plasma concentrations occurred in some dogs54,55 and one the individual response, buprenorphine SR-LAB adminis-
pig,49 indicating that there may be individual absorption tered at 1.8 mg/kg IM to American kestrels would require
variability with these patches as well. More research and use administration every 24 hours to manage pain.61
of these patches is indicated in zoological medicine. In red-tailed hawks (Buteo jamaicensis), the pharma-
cokinetics of two dosages of a concentrated formulation
Avian of buprenorphine (Simbadol) were evaluated after SC
In a pharmacokinetic study in American kestrels (Falco administration.58 Maximum buprenorphine concentration
sparverius), SC and intramuscular (IM) administration was achieved at 5 and 15 minutes for the 0.3 mg/kg and
of 1.8 mg/kg of buprenorphine SR-LAB were both char- 1.8 mg/kg doses, and plasma concentrations were main-
acterized by rapid absorption and elimination kinetics.57 tained at greater than 1 ng/mL for at least 24 and 48 hours,
CHAPTER 26 Sustained-Release and Long-Acting Opioid Formulations of Interest in Zoological Medicine 161
fentanyl in cats anesthetized with isoflurane, Am J Vet Res 37. Pergolizzi J, Aloisi AM, Dahan A, et al: Current knowledge of
64:1557–1561, 2003. buprenorphine and its unique pharmacological profile, Pain Pract
21. Wilson D, Pettifer GR, Hosgood G: Effect of transdermally 10:428–450, 2010.
administered fentanyl on minimum alveolar concentration of 38. Catbagan DL, Quimby JM, Mama KR, et al: Comparison of
isoflurane in normothermic and hypothermic dogs, J Am Vet Med the efficacy and adverse effects of sustained-release buprenor-
Assoc 228:1042–1046, 2006. phine hydrochloride following subcutaneous administration
22. Mills PC, Magnusson BM, Cross SE: Investigation of in vitro and buprenorphine hydrochloride following oral transmucosal
transdermal absorption of fentanyl from patches placed on skin administration in cats undergoing ovariohysterectomy, Am J Vet
samples obtained from various anatomic regions of dogs, Am J Res 72:461–466, 2011.
Vet Res 65:1697–1700, 2004. 39. Nunamaker EA, Stolarik DF, Ma J, et al: Clinical efficacy of
23. Deschamps J-Y, Gaulier J-M, Podevin G, et al: Fatal overdose sustained-release buprenorphine with meloxicam for postopera-
after ingestion of a transdermal fentanyl patch in two non-human tive analgesia in beagle dogs undergoing ovariohysterectomy, J
primates, Vet Anaesth Analg 39:653–656, 2012. Am Assoc Lab Anim Sci 53:494–501, 2014.
24. Freise KJ, Linton DD, Newbound GC, et al: Population phar- 40. DiVincenti L, Meirelles LAD, Westcott RA: Safety and clinical
macokinetics of transdermal fentanyl solution following a single effectiveness of a compounded sustained-release formulation
dose administered prior to soft tissue and orthopedic surgery in of buprenorphine for postoperative analgesia in New Zealand
dogs, J Vet Pharmacol Ther 35:65–72, 2012. White rabbits, J Am Vet Med Assoc 248:795–801, 2016.
25. Linton DD, Wilson MG, Newbound GC, et al: The effective- 41. Seymour TL, Adams SC, Felt SA, et al: Postoperative analgesia
ness of a long-acting transdermal fentanyl solution compared to due to sustained-release buprenorphine, sustained-release meloxi-
buprenorphine for the control of postoperative pain in dogs in cam, and carprofen gel in a model of incisional pain in rats
a randomized, multicentered clinical study, J Vet Pharmacol Ther (Rattus norvegicus), J Am Assoc Lab Anim Sci 55:300–305, 2016.
35:53–64, 2012. 42. Chum HH, Jampachairsri K, McKeon GP, et al: Antinocicep-
26. Martinez SA, Wilson MG, Linton DD, et al: The safety and tive effects of sustained-release buprenorphine in a model of
effectiveness of a long-acting transdermal fentanyl solution incisional pain in rats (Rattus norvegicus), J Am Assoc Lab Anim
compared with oxymorphone for the control of postoperative Sci 53:193–197, 2014.
pain in dogs: a randomized, multicentered clinical study, J Vet 43. Kendall LV, Hansen RJ, Dorsey K, et al: Pharmacokinetics of
Pharmacol Ther 37:394–405, 2014. sustained-release analgesics in mice, J Am Assoc Lab Anim Sci
27. Freise KJ, Newbound GC, Tudan C, et al: Naloxone reversal of 53:478–484, 2014.
an overdose of a novel, long-acting transdermal fentanyl solution 44. Kendall LV, Wegenast DJ, Smith BJ, et al: Efficacy of sustained-
in laboratory Beagles, J Vet Pharmacol Ther 35:45–51, 2012. release buprenorphine in an experimental laparotomy model in
28. Savides MC, Pohland RC, Wilkie DA, et al: The margin of safety female mice, J Am Assoc Lab Anim Sci 55:66–73, 2016.
of a single application of transdermal fentanyl solution when 45. Healy JR, Tonkin JL, Kamarec SR, et al: Evaluation of an
administered at multiples of the therapeutic dose to laboratory improved sustained-release buprenorphine formulation for use
dogs, J Vet Pharmacol Ther 35:35–43, 2012. in mice, Am J Vet Res 75:619–625, 2014.
29. Carlson AM, Kelly R, Fetterer DP, et al: Pharmacokinetics of 2 46. Smith BJ, Wegenast DJ, Hansen RJ, et al: Pharmacokinetics and
formulations of transdermal fentanyl in cynomolgus macaques paw withdrawal pressure in female guinea pigs (Cavia porcellus)
(Macaca fascicularis), J Am Assoc Lab Anim Sci 55:436–442, treated with sustained-release buprenorphine and buprenor-
2016. phine hydrochloride, J Am Assoc Lab Anim Sci 55:789–793,
30. Salyards GW, Lemoy MJ, Knych HK, et al: Pharmacokinetics 2016.
of a novel, transdermal fentanyl solution in rhesus macaques 47. Cary CD, Lukovsky-Akhsanov NL, Gallardo-Romero NF, et al:
(Macaca mulatta), J Am Assoc Lab Anim Sci 2017. Pharmacokinetic profiles of meloxicam and sustained-release
31. Kottwitz J, Boothe M, Harmon R, et al: Results of the megaver- buprenorphine in prairie dogs (Cynomys ludovicianus), J Am Assoc
tebrate analgesia survey: elephants and rhino, J Zoo Wildl Med Lab Anim Sci 2017.
47:301–310, 2016. 48. Dooley SB, Aarnes TK, Lakritz J, et al: Pharmacokinetics and
32. Delaski KM, Gehring R, Heffron BT, et al: Plasma concen- pharmacodynamics of buprenorphine and sustained-release
trations of fentanyl achieved with transdermal application in buprenorphine after administration to adult alpacas, Am J Vet
chickens, J Avian Med Surg 31:6–15, 2016. Res 78:321–329, 2017.
33. Waugh L, Knych H, Cole G, et al: Pharmacokinetic evaluation 49. Thiede AJ, Garcia KD, Stolarik DF, et al: Pharmacokinetics
of a long-acting fentanyl solution after transdermal administra- of sustained-release and transdermal buprenorphine in göttin-
tion in helmeted guineafowl (Numida meleagris), J Zoo Wildl Med gen minipigs (Sus scrofa domestica), J Am Assoc Lab Anim Sci
47:468–473, 2016. 53:692–699, 2014.
34. Darrow BG, Myers GE, KuKanich B, et al: Fentanyl transdermal 50. Walkowiak KJ, Graham ML: Pharmacokinetics and antinocicep-
therapeutic system provides rapid systemic fentanyl absorption in tive activity of sustained-release buprenorphine in sheep, J Am
two ball pythons (Python regius), J Herpetol Med Surg 26:94–99, Assoc Lab Anim Sci 54:763–768, 2015.
2016. 51. Nunamaker EA, Halliday LC, Moody DE, et al: Pharmacokinet-
35. Kharbush RJ, Gutwillig A, Hartzler KE, et al: Antinociceptive ics of 2 formulations of buprenorphine in macaques (Macaca
and respiratory effects following application of transdermal fen- mulatta and Macaca fascicularis), J Am Assoc Lab Anim Sci
tanyl patches and assessment of brain mu-opioid receptor mRNA 52:48–56, 2013.
expression in ball pythons, Am J Vet Res 78:785–795, 2017. 52. Molter CM, Barbosa L, Johnson S, et al: Pharmacokinetics of
36. Gamble K: Plasma fentanyl concentrations achieved after trans- a single subcutaneous dose of sustained release buprenorphine
dermal fentanyl patch application in prehensile-tailed skinks, in northern elephant seals (Mirounga angustirostris), J Zoo Wildl
Corucia zebrata, J Herp Med Surg 18:81–85, 2008. Med 46:52–61, 2015.
CHAPTER 26 Sustained-Release and Long-Acting Opioid Formulations of Interest in Zoological Medicine 163
53. Moll X, Fresno L, García F, et al: Comparison of subcutaneous 64. Malmlov A, Campbell T, Monnet E, et al: Diagnosis, surgical
and transdermal administration of buprenorphine for pre-emptive treatment, recovery, and eventual necropsy of a leopard (Panthera
analgesia in dogs undergoing elective ovariohysterectomy, Vet J pardus) with thyroid carcinoma, Case Rep Vet Med 2014:2014.
187:124–128, 2011. 65. McNulty EE, Gilson SD, Houser BS, et al: Treatment of
54. Andaluz A, Moll X, Ventura R, et al: Plasma buprenorphine fibrosarcoma in a maned wolf (Chrysocyon brachyurus) by rostral
concentrations after the application of a 70 microg/h transder- maxillectomy, J Zoo Wildl Med 31:394–399, 2000.
mal patch in dogs. Preliminary report, J Vet Pharmacol Ther 66. Gjeltema JL, MacLean RA, Cohen EB, et al: Hypertrophic
32:503–505, 2009. osteodystrophy in two red wolf (Canis rufus) pups, Case Rep Vet
55. Pieper K, Schuster T, Levionnois O, et al: Antinociceptive effi- Med 2015:2015.
cacy and plasma concentrations of transdermal buprenorphine in 67. Spriggs M, Arble J, Myers G: Intervertebral disc extrusion and
dogs, Vet J 187:335–341, 2011. spinal decompression in a binturong (Arctictis binturong), J Zoo
56. Murrell JC, Robertson SA, Taylor PM, et al: Use of a transdermal Wildl Med 38:135–138, 2007.
matrix patch of buprenorphine in cats: preliminary pharmacoki- 68. Naples LM, Lacasse C, Landolfi JA, et al: Acute pancreatitis
netic and pharmacodynamic data, Vet Rec 160:578–583, 2007. in slender-tailed meerkats (Suricata suricatta), J Zoo Wildl Med
57. Guzman DS-M, Knych HK, Olsen GH, et al: Pharmacokinet- 41:275–286, 2010.
ics of a sustained release formulation of buprenorphine after 69. Wiedner E, Holland J, Trupkiewicz J, et al: Severe laminitis in
intramuscular and subcutaneous administration to American multiple zoo species, Vet Q 34:22–28, 2014.
kestrels (Falco sparverius), J Avian Med Surg 31:102–107, 2017. 70. Howard LL, Richardson GL: Transposition of the biceps tendon
58. Gleeson M, Guzman DS, Kass PH, et al: Pharmacokinetics of to reduce lateral scapulohumeral luxation in three species of
a concentrated buprenorphine formulation in red-tailed hawks nondomestic ruminant, J Zoo Wildl Med 36:290–294, 2005.
(Buteo jamaicensis), Am J Vet Res 79(1):13–20, 2018. 71. Savageau NR, Gamble KC: Clinical challenge: renal adenocarin-
59. Laniesse D, Guzman DS, Knych HK, et al: Pharmacokinetics of coma in a beaded lizard (Heloderma horridum horridum), J Zoo
butorphanol tartrate in a long-acting poloxamer 407 gel formu- Wildl Med 47:945–947, 2016.
lation administered to Hispaniolan Amazon parrots (Amazona 72. Spriggs M, Thompson KA, Barton D, et al: Gastrolithiasis in
ventralis), Am J Vet Res 78:688–694, 2017. prehensile-tailed porcupines (Coendou prehensilis): nine cases and
60. Clancy MM, KuKanich B, Sykes IVJM: Pharmacokinetics of pathogenesis of stone formation, J Zoo Wildl Med 45:883–891,
butorphanol delivered with an osmotic pump during a seven- 2014.
day period in common peafowl (Pavo cristatus), Am J Vet Res 73. Simeone CA, Colitz CMH, Colegrove KM, et al: Subconjunctival
76:1070–1076, 2015. antimicrobial poloxamer gel for treatment of corneal ulceration
61. Sanchez-Migallon Guzman D, Ceulemans S, Olsen GH, et al: in stranded California sea lions (Zalophus californianus), Vet
Evaluation of the thermal antinociceptive effects of a sustained- Ophthalmol 20(5):441–449, 2017.
release buprenorphine formulation after intramuscular adminis- 74. Kline S, Rooker L, Nobrega-Lee M, et al: Hypoadrenocorticism
tration to American kestrels (Falco sparverius), Journal of Avian (Addison’s disease) in a hoffmann’s two-toed sloth (Choloepus
Medicine and Surgery (in press). hoffmanni), J Zoo Wildl Med 46:171–174, 2015.
62. Galloway DS, Coke RL, Rochat MC, et al: Spinal compres- 75. Marrow J, Viner T, Thompson R, et al: Uterine adenomyosis
sion due to atlantal vertebral malformation in two african lions in southern three-banded armadillos (Tolypeutes matacus), J Zoo
(Panthera leo), J Zoo Wildl Med 33:249–255, 2002. Wildl Med 44:1018–1026, 2013.
63. Herrin KV, Allan G, Black A, et al: Stifle osteochondritis dissecans
in snow leopards (Uncia uncia), J Zoo Wildl Med 43:347–354,
2012.
27
Use of Naltrexone and Atipamezole
in Emergency Response to Human
Exposure to Ultra-Potent Opioids
and Alpha-2 Agonists in Zoo and
Wildlife Medicine
JEFFERY R. ZUBA AND MARK GREENBERG
164
CHAPTER 27 Use of Naltrexone and Atipamezole in Emergency Response to Human Exposure to Ultra-Potent Opioids 165
TABLE Equipotency Data of Clinically Significant Doses of Morphine and Ultra-Potent Opioids Used in Zoo and
27.1 Wildlife Anesthesia
Note: The morphine dose provided is known to cause respiratory depression in humans. The volume of UPO necessary to provide this hypothetical accidental
exposure is also listed.
*Morphine IV dose causing respiratory depression in 65 kg opioid naïve human (see Refs. 16 and 42).
†
Volume (mL) equals equipotent dose (mg) times the UPO concentration (mg/mL).
‡
Carfentanil equipotency to morphine, 10,000:1 (see Refs. 11 and 12).
§
20 mg morphine divided by 10,000.
**Etorphine equipotency to morphine, 6000:1 (see Ref. 2).
††
20 mg morphine divided by 6000.
‡‡
Thiafentanil equipotency to morphine, 6000:1 (see Ref. 10).
§§
20 mg morphine divided by 6000.
IV, Intravenous; UPO, ultra-potent opioids.
widely used UPO in zoo and wildlife anesthesia,5 and is Etorphine, Thiafentanil, and Carfentanil
the induction agent of choice for elephant, rhinoceros, Potency Comparisons
nondomestic equids, and other hoofstock. It is often Carfentanil was considered the most potent of the UPOs
combined with azaperone, medetomidine, midazolam, or in most zoo and wildlife species, followed by thiafentanil
azaperone to produce muscle relaxation.6 The availability of and then etorphine. On a mg:mg basis, a rough estimate
etorphine since its first use and description in the late 1960s of clinical equipotency for most captive ungulate species
revolutionized the ability of veterinarians to safely capture is 1 mg carfentanil to 1.75 mg thiafentanil to 2.0–2.5 mg
and restrain many species that previously could not be etorphine.4 Although somewhat variable in the veterinary
handled.2 literature, etorphine and thiafentanil are estimated to be
6000 times more potent than morphine,2,10 and carfentanil
Thiafentanil 10,000 times more potent.11,12 See Table 27.1 for a review
Thiafentanil (Thianil, 10 mg/mL, Wildlife Pharmaceuticals, of equipotency data.
Inc.), previously known as A-3080, was introduced in the
early 1990s and has similar characteristics to etorphine and Butorphanol
carfentanil, but with faster inductions in certain species.2,7 Butorphanol is a mixed opioid agonist-antagonist com-
Since thiafentanil has a shorter half-life than etorphine and monly used in domestic and nondomestic veterinary
carfentanil, there is less chance for renarcotization, which species for analgesia, sedation, or improved quality of
is particularly important in free-ranging wildlife.7 Some anesthesia.6,13,14 It is estimated to be four to seven times
studies have shown little advantage of thiafentanil as an more potent than morphine but has not been considered
immobilizing agent over other UPOs in most species of to be a UPO. However, it is mentioned here because it is
ungulates.8,9 Its use in zoo and wildlife will likely increase now available in a concentrated solution (50 mg/mL, Wild-
due to the recent removal of carfentanil from production. life Pharmaceuticals, Inc.) and as a constituent of BAM
It is often combined with supplemental drugs to produce (Wildlife Pharmaceuticals, Inc.), a commercially available
balanced anesthesia. A comprehensive review of this drug combination of butorphanol, azaperone, and medetomi-
with dose recommendations is available in zoo and wildlife dine, which is reviewed below. A thorough review of the
species.7 utility of butorphanol in zoo and wildlife species is found
in the literature.13
Carfentanil
Carfentanil was the most potent of the UPOs and, in Opioid Antagonists
general, had similar immobilizing properties as etorphine
and thiafentanil in ungulates.1,2,5 It should be noted that Naloxone is a short-acting opioid antagonist and the
as of 2016, carfentanil was no longer available from the drug of choice for reversal of acute opioid intoxication
manufacturer (Wildlife Pharmaceuticals, Inc.). It was a in humans.14–16 It is commonly found in zoo and wildlife
commonly used UPO in the United States in a variety of veterinarians’ emergency response kit in case of acciden-
zoo and wildlife ungulate species for nearly 30 years. tal human exposure. Due to its short half-life (30–60
166 SE C T I O N 6 Anesthesia and Analgesia
minutes),16 it is not used to reverse the effects of UPOs in Other Alpha-2 Agonists
zoo and wildlife, which have considerably longer durations
of action. Naltrexone is a long-acting pure opioid antagonist Detomidine is an injectable α-2 agonist available and widely
and a relative of naloxone.6,14 It is the reversal drug of choice used, especially in domestic horses.19 It does not come in a
for the long-acting UPOs.7 Nalmefene is another long- highly concentrated form like medetomidine, but it is still
acting opioid antagonist that has been studied in captive a very useful drug in zoo and wildlife anesthesia. Xylazine
ungulates but is not commercially available and its half-life is now available in a highly concentrated injectable solution
is shorter than naltrexone.6,16,17 (300 mg/mL, Wildlife Pharmaceuticals, Inc.), which allows
it to be used in darts combined with other drugs for remote
Ultra-Potent Alpha-2 Agonists delivery. This also adds to the danger to humans at high
and Antagonists doses.27
TABLE
27.2 Hypothetical Exposures to Dangerous Drugs Used in Zoo and Wildlife Anesthesia
Is This a
Significant Volume
of Exposure?
Dose of Dose of Significance Is
Spray Etorphine Thiafentanil Dose of Dose of Expressed as
Needle Droplet (10 mg/mL) (10 mg/mL) Carfentanil Medetomidine Multiple of Known
Type of Volume* Volume† in Volume in Volume (3 mg/mL) in (40 mg/mL) in Harmful Dose of
Exposure (mL) (mL) (mg) (mg) Volume (mg) Volume (mg) Morphine‡
27 ga × 0.00044§ 0.0044 0.0044 0.00132 0.018 [0.13]§§ Etorphine, 1.3 times
1
2 in. [0.0033]** [0.0033]†† [0.002]‡‡ Thiafentanil, 1.3 times
needle Carfentanil, 0.66 times
stick Medetomidine, 0.14
times
22 ga × 0.00343*** 0.0343 0.0343 0.0103 1.37 [0.13]§§ Etorphine, 10.4 times
1 in. [0.0033]** [0.0033]†† [0.002]‡‡ Thiafentanil, 10.4
needle times
stick Carfentanil, 5.2 times
Medetomidine, 10.5
times
Spray 0.05 0.5 0.5 0.15 [0.002]‡‡ 2.0 [0.13]§§ Etorphine, 152 times
droplet [0.0033]** [0.0033]†† Thiafentanil, 152 times
Carfentanil, 75 times
Medetomidine, 15.4
times
Note: Estimated volumes and mg dose of ultra-potent opioids or medetomidine from a hypothesized needle stick or spray droplet exposure. Clinical significance
of the dose is estimated from comparative data in the literature for morphine, ultra-potent opioids, medetomidine, and dexmedetomidine.
*Volume of a cylinder = πr2h
• Refer http://www.math.com/tables/geometry/volumes.htm
• π = 3.14, r is the radius of needle lumen, h is the length of needle lumen
†
Volume of a droplet = 0.05 mL, http://www.endmemo.com/sconvert/milliliterdrop.php.
‡
Estimated dose of morphine that causes respiratory depression is 20 mg IV (see Refs. 16 and 42).
§
Volume of a 27 ga × 12 in. needle = πr2h
• Refer http://www.sigmaaldrich.com/chemistry/stockroom-reagents/learning-center/technical-library/needle-gauge-chart.html
• π = 3.14, r is the radius = 0.105 mm, h is length = 12.7 mm
• Therefore: (3.14) (0.105 mm)2 (12.7 mm) = 0.44 mm3 or 0.44 µL or 0.00044 mL
**Estimated dose of Etorphine that would cause respiratory depression using morphine equipotency in human (see Table 27.1).
††
Estimated dose of Thiafentanil that would cause respiratory depression using morphine equipotency in human (see Table 27.1).
‡‡
Estimated dose of Carfentanil that would cause respiratory depression using morphine equipotency in human (see Table 27.1).
§§
Estimated dose of Medetomidine, (2 µg/kg, 65 kg human, 0.13 mg dose) that would cause cardiovascular symptoms using dexmedetomidine equipotency
in humans (see Refs. 49 and 59).
***Volume of a 22 ga × 1 in. needle = πr2h,
• Refer http://www.sigmaaldrich.com/chemistry/stockroom-reagents/learning-center/technical-library/needle-gauge-chart.html
• π = 3.14, r is the radius = 0.2065 mm, h is length = 25.4 mm
• Therefore: (3.14) (0.2065)2 (25.4 mm) = 3.43 mm3 or 3.43 µL or 0.00343 mL.
due to distractions, carelessness, inexperience, procedural Aerosolization of potent anesthetic agents may occur due
pressures, or challenging environmental conditions. to spray exposure resulting in direct contact with skin or
mucous membranes. Transdermal and transmucosal absorp-
Routes of Exposure tion of certain drugs, such as fentanyl, is well documented
and capitalized upon in the development of human and
Accidental injection is the most obvious exposure and must veterinary drug delivery routes. This type of absorption
be dealt with as life-threatening. In one survey, needle stick depends on the concentration and amount of drug; dura-
exposures in zoo veterinarians were reported as high as tion, location, and surface area of exposure; temperature
87%.29 Significantly, 17.2% of those exposures were while and integrity of the skin or mucosa; and lipophilicity,
working with immobilizing agents. The most catastrophic molecular weight, and solubility.30,31 Fentanyl and most of
type of accidental exposure would be a deep IM injec- its derivatives are highly lipid soluble and are known to be
tion due to a dart hitting a human. See Tables 27.2 and absorbed across the skin.2,32,33 Presumably, all of our UPOs
27.3 for hypothetical exposure of this type of accidental would have similar disposition. In human medicine, trans-
injection. dermal and transmucosal fentanyl are used for analgesia
168 SE C T I O N 6 Anesthesia and Analgesia
TABLE Hypothetical Scenarios With Known Accidental Human Exposure to Potent Anesthetic Agents and
27.3 Suggested Response With Available Antagonists
AT, Atipamezole; AZAP, azaperone; BUT, butorphanol; ETOR, etorphine; FLU, flumazenil; IM, intramuscular; IN, intranasal; IV, intravenous; KET, ketamine; MED,
medetomidine; MIDAZ, midazolam; NAL, naloxone; TREX, naltrexone; UPO, ultra-potent opioid.
Note: In these cases, the veterinarian justifiably provides immediate action due to dire, life-threatening circumstances and lack of options. Patient may be exhibiting
symptoms or they are expected. Please refer to Figs. 27.1–27.3 for emergency response algorithms. The reader must understand this table is hypothetical,
and authors cannot recommend the use of antagonists but provide this information based on evidence to support its consideration in a known emergency.
by skin patch, lozenge, buccal patch, and lollipops.32,33 In The clinically significant or lethal dose of the UPOs in
zoo and wildlife species, transmucosal delivery of UPOs humans is unknown, but if we use the estimated analgesic
for anesthesia and sedation has been reported in the black potency of these drugs referenced in the literature,1,2,38 we
bear34 and brown bear35,36 and UPOs and α-2 agonists can create hypothetical examples. Morphine is considered
in a tapir.37 Therefore the literature supports transdermal the gold standard for comparing analgesic potency of opioids
and transmucosal absorption as a predictable method of and other analgesics.39–41 The clinically significant dose of
controlled, and likely accidental, administration of potent intravenous (IV) morphine causing respiratory depres-
opioids and α-2 agonists. See Tables 27.2 and 27.3 for sion in the opioid naïve 65 kg human is approximately
hypothetical exposure of this type of accidental injection. 20 mg.41,42 Doses higher than this are expected to become
more severe and life-threatening. Therefore a clinically equi-
Significance of an Exposure potent morphine dose for etorphine and thiafentanil (6000
× morphine) would be approximately 3.3 µg, whereas the
Determining what constitutes a clinically significant expo- carfentanil (10,000 × morphine) equipotent dose would be
sure is difficult, but for safety purposes, we must assume only 2.0 µg. See Table 27.2 for hypothetical examples dem-
that any accidental exposure should elicit an emergency onstrating the extremely small volumes of each UPO that
response. Fortunately, significant exposures appear to be may cause respiratory depression in an accidental exposure.
rare and are attributed to redundancy in safety measures, It must be restated that the potency of the UPOs presented
training, and careful attention to detail by the attending here originate from referenced extrapolated analgesia data
veterinarian.1,29 Minor exposures are more probable but are but then provides the only data available to predict the
likely not to be reported in the literature.2,38 dangers of the UPOs.
CHAPTER 27 Use of Naltrexone and Atipamezole in Emergency Response to Human Exposure to Ultra-Potent Opioids 169
Mathematics of Exposures dissociative agents), then care of the victim will be sup-
portive using the principles of Basic Life Support (BLS)54
To better understand the clinical significance of an exposure until medical transport arrives. If the anesthetic agent
to a UPO or concentrated medetomidine, a hypothetical has a pharmacologic antagonist and one is available, the
example with quantification of the exposure is necessary. next immediate steps will be to consider administering
Because minor exposures to these drugs are more likely, a the antagonist (see Fig. 27.1) and securing IV access. For
situation is presented where a person is accidentally exposed victims exposed to a dangerous drug in a remote area and
by a needle stick or a minor spray from a mishandled far from medical care, a rapid response will be needed to
dart or syringe. A more dangerous situation is possible prevent a fatality. For benzodiazepines, flumazenil will be
with exposure to larger amounts or a combination of these helpful in most cases. In the next few sections, specific
potent drugs. The consequence of a human exposure to medical response to individual agents will be addressed.
medetomidine will be compared with clinically significant
doses of dexmedetomidine used in human anesthesia. This Ultra-Potent Opioids: Etorphine, Carfentanil,
information is reviewed in Tables 27.1–27.3. and Thiafentanil
Needle Stick Exposure Due to extremely high potency, needle stick accidents con-
Needle stick exposures by zoo and wildlife veterinarians taining etorphine, and presumably the other UPOs, have
are a potential source of accidental injection of dangerous resulted in respiratory arrest.38 As a safety measure, strict
drugs.29,38,43 To demonstrate, we pose a situation in which protocols are in place for veterinarians who handle these
a person accidentally sticks himself or herself with a needle dangerous drugs.1,3,4,55 However, despite careful handling
with an unpressurized syringe. Only the volume of the practices, accidents do occur, and thus an UPO exposure
drug found in the lumen of the needle is hypothesized to treatment protocol is necessary (see Fig. 27.2). Having an
be injected. The lumen volume of a needle is estimated opioid reversal kit that includes airway management equip-
to be the volume of a cylinder, as per the mathematical ment, IV placement kit, and enough naloxone for a 100-kg
equation πr2h, where π = 3.14, r is the radius of the needle’s patient may be lifesaving.3
lumen, and h is the needle length. A needle gauge chart When exposure of a human to a UPO occurs, the first
is then used to determine the radius and length (http:// step is to notify other personnel and activate the EMS
www.sigmaaldrich.com/chemistry/stockroom-reagents/ by dialing “911” or the appropriate local emergency alert
learning-center/technical-library/needle-gauge-chart.html). system. Of all the dangerous anesthetic agents, UPO
Veterinarians commonly use 27-ga × 1 2 in. (from a TB exposure has the potential to cause a fatality the fastest
syringe) and 22-ga × 1 in. needles to withdraw small and with only a seemingly minor exposure dose. An opioid
volumes of concentrated drugs, and these will be used antagonist is the antidote to reverse the central nervous
as examples in this hypothetical exposure. Results are system and respiratory depression effects. If time allows,
found in Tables 27.2 and 27.3 and represent only a minor attach monitors and obtain IV access. The patient should
exposure of a needle stick with the volume found within not be left unattended while waiting for EMS to arrive.
the lumen. If the patient is already unconscious, consider immediate
administration of an opioid antagonist. If the victim is not
Spray Exposure breathing, open the airway and begin rescue breathing with
Aerosolization of these drugs with exposure to the skin or a bag and mask or using mouth-to-mouth ventilation.
mucosa may occur by mishandling a syringe or dart or in the With UPO exposure, respiratory depression leading to
event of an accident. In this example, we pose a hypothetical apnea will be the principal clinical symptom, with the effects
situation where a person is exposed to a single droplet of a resulting in hypoxia and death. The primary resuscitative
potent anesthetic solution. This may easily be amplified, of measure for UPO overdose is administration of an opioid
course, if exposed to numerous droplets. The volume of a antagonist. Naloxone is a competitive mu opioid–receptor
fluid droplet is estimated to be 50 µl, or 0.05 mL (http:// antagonist that reverses all signs of opioid intoxication. It
www.endmemo.com/sconvert/milliliterdrop.php). Results is typically supplied as a 0.4 mg/mL or a 10 mL multidose
from this type of exposure by various drugs and proposed vial of 1 mg/mL formulation and can be given IM, IV,
clinical significance can be found in Tables 27.2 and 27.3. subcutaneously (SC), intranasally (IN), or intratracheally
(IT).32,41 The initial dose of naloxone for a victim who is
Medical Management for Accidental symptomatic should be a 5 mg naloxone IV push.56 If there
is no IV in place, then naloxone should be given IM. For
Veterinary Anesthetic Exposure a significant UPO exposure, the effective dose may need to
Agent-Specific Resuscitation Protocols be much higher and repeated every 2–3 minutes, depending
on the clinical picture.38 The onset of action of naloxone is
After initial evaluation of the victim, the next steps of the less than 2 minutes when administered intravenously.44 The
resuscitation will depend on which agents are involved (Fig. effective duration of action is only 20–30 minutes, which
27.1). If the exposure agent has no antagonist (tranquilizers, may be much shorter than that of a UPO. Thus, after
170 SE C T I O N 6 Anesthesia and Analgesia
Monitor
Yes Significant exposure No Observe
Consider transfer to
medical facility
Monitor
Observe
Yes Is the victim symptomatic? No Start IV
Facemask O2
Transfer to hospital
Dial 911
Flush wound with water
Place on monitors
Obtain IV access
Facemask O2, 10 L/min
Prepare for airway management
Immediate transfer to medical facility
Continue supportive care
ACLS protocols should guide treatment
‡
Opioids Alpha-2 agonists Benzodiazepines
(etorphine, thiafentanil, (medetomidine, xylazine, detomidine, (diazepam, midazolam)
carfentanil) dexmedetomidine)
0.5 mg IV flumazenil
5 mg naloxone IV/IM Consider 0.3 mg/kg IV atipamezole
‡
Repeat every 2–3 min as needed Consider 0.6 mg/kg IM atipamezole These agents are not usually potent enough
Repeat every 2–3 min as needed to cause severe symptoms but may potentiate
Consider 100 mg naltrexone IM other agents, where its reversal will be lifesaving.
Consider 100 mg atipamezole IM
• Figure 27.1 Algorithm for management of accidental exposure to anesthetic agents. ACLS, Advanced
Cardiac Life Support; CNS, central nervous system; IM, intramuscular; IV, intravenous.
stabilization, intensive patient monitoring will be needed for sulfate and naltrexone hydrochloride, extended-release cap-
all significant UPO exposures watching for re-narcotization. sules, Pfizer, Inc., New York), as a failsafe to prevent opioid
An algorithm for UPO accidental exposure and overdose is abuse.57 Naltrexone has opioid antagonist effects when
presented in Fig. 27.2. In critical cases, a significant concern given orally and has been used IM in humans in emergency
will be the availability of sufficient opioid antagonist. situations.38 If an accident occurs in a remote area, it is
Naltrexone is a long-acting opioid antagonist and unlikely naloxone will be available in sufficient quantities
another potential option for UPO exposures. It is routinely to antagonize a long acting UPO. In a reported case of
used in zoo and wildlife anesthesia to reverse UPO during carfentanil overdose, up to 50 mg of naltrexone was neces-
immobilization of megavertebrates and other ungulate sary to keep the patient breathing.38 Therefore naltrexone
species. Naltrexone is used in human medicine primarily 50–100 mg IM, which should be available for reversal of
as an oral agent to treat opioid addiction or for IM use in the anesthesia for the animal, would be expected to reverse
depot form with monthly injections.46 It is not currently significant opioid toxicity. Due to its proven safety in
available for emergency parenteral use in humans. It has humans, 50–100 mg IM naltrexone should be strongly
been studied in humans as part of EMBEDA (morphine considered in emergency situations where other therapy
CHAPTER 27 Use of Naltrexone and Atipamezole in Emergency Response to Human Exposure to Ultra-Potent Opioids 171
Ocular
Dart injection
Exposure incident Cutaneous
Victim unconscious
Significant exposure Less significant Needle stick
Victim apneic
Unstable but potentially harmful exposure No immediate
symptoms
Yes No
Naloxone
is not available Patient develops symptoms
Admit to ICU
Monitor oxygen saturation
*ACLS protocols should guide treatment
Positive pressure ventilation †
Large exposures may require large doses
Consider intubation
(greater than 10 mg) of Naloxone. Supplied
as 1 mg/mL, stock 10 mL vials
‡
Consider naltrexone§ Naloxone can be given intramuscularly (IM),
One should consider the portion subcutaneously (SC), or intranasally (IN)
§
of pathway in green if critical care If there is no other alternative, consider
is not immediately available or if using reversal for animal. Naltrexone
naloxone is unavailable or ineffective Zoopharm 50 mg/mL. Avoid Revivon
• Figure 27.2 Opioid resuscitation pathway. ACLS, Advanced Cardiac Life Support; ICU, intensive care
is ineffective or not available. Due to its long duration of Alpha-2 Agonists: Medetomidine and
action, re-narcotization following naltrexone use in animals Dexmedetomidine
is uncommon, making naltrexone a potentially attractive
option in a UPO emergency. Naltrexone, like naloxone, will Medetomidine, a highly selective α-2 agonist, is currently
cause opioid withdrawal in opioid-dependent patients.46,58 It available in a highly concentrated form (40 mg/mL, Wildlife
should be noted that diprenorphine ([Revivon, VetaPharma Pharmaceuticals, Inc.) for use in zoo and wildlife species.
Ltd., Leeds, UK], a reversal agent that comes packaged with In comparison, dexmedetomidine for human use (Precedex,
some formulations of etorphine; Immobilon, VetaPharma, Pfizer, Inc.) is available in a less potent formulation of
Ltd.) should not be given to a human as an antagonist due 100 mcg/mL. Medetomidine is approximately 50% as
to further depression of the patient.38 Diprenorphine has potent as dexmedetomidine.6,18,19 Therefore, in comparison
agonist-antagonist properties, which may be responsible for with human dexmedetomidine, this veterinary formula-
its lack of efficacy in UPO overdose. Thus, when working tion of medetomidine is approximately 20,000 times more
with any UPO, we recommend having an adequate supply concentrated on a per mL basis. In one report, dexmedeto-
of naloxone or parenteral naltrexone available for emergency midine was found to cause cardiac arrest at doses of 1–2
resuscitation. mcg/kg in humans.59 IM dexmedetomidine at 2.5 mcg/kg
172 SE C T I O N 6 Anesthesia and Analgesia
Consider atipamezole
Admit to ICU
Consider invasive blood pressure monitoring
Consider norepinephrine infusion for continued
or recurrent cardiovascular compromise
*ACLS protocols should guide treatment
†
One should consider the portion of pathway in green Bring Atipamezole and a copy of the protocol
if critical care is not immediately available or if with the victim to the ED. Do not exceed 100 mg/dose
‡
standard ACLS measures fail Atipamezole is supplied as 25 mg/mL 10 mL vial
• Figure 27.3 α-2 agonist resuscitation pathway. ACLS, Advanced Cardiac Life Support; ICU, intensive
care unit; IM, intramuscular; IV, intravenous; MED, medetomidine.
provided sedation in human trial and was reversed with and tolazoline have also been used for α-2 agonist over-
atipamezole.49 Medetomidine would be expected to have dose, but tolazoline is a nonspecific α-inhibitor, and there
sedative hypnotic effects in humans in quantities as small is no parenteral preparation of yohimbine available for
as 100 mcg, which is only 20 µL.49 In contrast to exposure human use.64,65
with an UPO that primarily causes respiratory depression There is no FDA-approved α-2 specific antagonist avail-
and apnea, α-2 agonist overdose is likely to result in severe able for parenteral use in humans. However, atipamezole, a
bradycardia.60,61 In these cases, at least initially, respiratory highly specific α-2 antagonist, is readily available for veteri-
drive may still be preserved; thus providing airway support nary use and is common in wildlife anesthesia. During an
alone will not likely be sufficient to prevent death. Cases of immobilization with medetomidine, or other α-2 agonist,
accidental and nonaccidental exposure have been reported, atipamezole usually would be available and prepared for
some with serious consequences.2 For practitioners of zoo use in the veterinary patient. Atipamezole has been tested
and wildlife medicine, accidental medetomidine overdose in dexmedetomidine sedated humans and was found to be
is a major concern.3,38 both safe and effective.49,66,67 Atipamezole, 100 mg IV, has
Treatment for accidental α-2 agonist overdose (cloni- been administered to anesthetized humans with minimal
dine, dexmedetomidine) in humans is generally supportive. side effects.66,67 It readily reverses dexmedetomidine
Treatment with naloxone and atropine has been tried, with sedation in humans when administered intravenously in
inconsistent results.62 Currently therapy consists of support- greater than a 40:1 ratio.49 It also increases norepinephrine
ing the victim with adrenergic agonists, activated charcoal, levels, in both anesthetized and dexmedetomidine-sedated
fluids, and treating respiratory failure with endotrache- humans. Using published data, it appears that humans
al intubation and mechanical ventilation.63 Yohimbine require approximately 10 times less α-2 agonist such as
CHAPTER 27 Use of Naltrexone and Atipamezole in Emergency Response to Human Exposure to Ultra-Potent Opioids 173
dexmedetomidine for deep sedation, but require 10 times practical perspective, if there is an antagonist for any of the
more atipamezole for adequate reversal when compared agents, it is most important to administer it quickly. An
with veterinary species.20 Although atipamezole is not FDA example would be if a human is exposed to high doses of
approved for use in humans, dose recommendations for concentrated midazolam, the rescue team should consider
emergency atipamezole use in humans do exist.20,21 (See administering flumazenil up to 500 mcg (0.5 mg) IV or IM
Fig. 27.3 for a resuscitation algorithm in the event of a as part of the resuscitation.68 Similarly, BAM is a combina-
significant α-2 agonist human exposure.) tion anesthetic that also could be very dangerous in an acci-
Using the data from human trials and anesthetic dental human exposure due to high doses of butorphanol
experience in primates and other animals, the α-2 agonist and medetomidine. Atipamezole and naltrexone should be
resuscitation pathway in Fig. 27.3 was developed. If the prepared as an antidote and immediately injected IM into
victim is stable, no immediate antagonist would be given. the victim. If administering the antagonist for one of the
The rescue team should monitor the victim for signs of agents doesn’t result in complete resolution of symptoms,
shock and prepare for transport. However, after a significant medical support of the victim may be required.
medetomidine exposure, especially in a remote area without
medical support, the rescuers should consider the use of ati-
pamezole as shown in Fig. 27.3. If the victim is unstable, Controversies in the Use of Antagonists in
100 mg atipamezole IM would be reasonable. If that is Human Exposure to Dangerous Drugs
not effective, it should be repeated. If the patient was
initially stable and then begins to develop symptoms, the A professional dilemma exists when presented with a medical
rescuers should consider giving atipamezole 0.6 mg/kg emergency involving a person with a known exposure to
IM or 0.3 mg/kg slow IV push. Because the dose of the UPO or α-2 agonists. Veterinarians are concerned about
α-2 agonist in an accidental exposure will not be precisely treating a human without a license. Emergency depart-
known, it seems more logical to administer atipamezole ment physicians in the United States do not have approval
using a mg/kg weight-based dose instead of a ratio of atipa- from federal regulating agencies, such as the US Food and
mezole: α-2 agonist (see Fig. 27.3). As with UPO, it is likely Drug Administration (FDA), to provide a patient with
the reversal agent for the animal being immobilized will be possibly the most pharmacologically appropriate antidote.
ready for use before the start of the procedure. If there is a During such an emergency, the gray, unclear boundaries
catastrophic medetomidine exposure and no other option of malpractice, ethics, and law must be considered. Legal
is available, it is reasonable to consider using the animals’ concerns are reviewed in the next section. The authors
reversal dose in the human victim. are careful not to go beyond the scope of our training
If the victim develops cardiac arrest, or the cardiac by making overconfident or unsubstantiated statements.
rhythm becomes nonperfusing, the rescuers should Rather, we provide a review of the most current literature
immediately start chest compressions. Adequate depth of on the use of naltrexone and atipamezole in humans as an
compressions during cardiopulmonary resuscitation (CPR) emergency treatment option based on scientific evidence,
will be essential to maintain perfusion to vital organs.8 If logic, and experience. Inevitably, it will be the decision of
available, atipamezole should be administered.20,21 Prompt the attending healthcare professional on how to proceed,
administration of the reversal agent in this situation may given the circumstance. It should be noted that the use of
reverse the cardiac arrest. Once at the hospital, ongoing any antagonist is not a substitute for seeking emergency
care will be dictated by Advanced Cardiac Life Support medical care for a person exposed to a dangerous anesthetic
(ACLS) protocols.68 The resuscitating physicians should agent.
consider implementing a norepinephrine infusion for severe
shock, as α-2 agonists lower norepinephrine levels.20,66,67 It Naltrexone Use in Humans
will be important for a member of the veterinary team to
accompany the patient with sufficient atipamezole and to Naltrexone and naloxone are pure opioid antagonists
bring the protocol in Fig. 27.3 along with the victim to the safe for use in humans. Naloxone has a short half-life of
hospital for the benefit of the emergency room physicians. approximately 60 minutes and is available in many forms
Human hospitals will not have atipamezole, and most of administration (IV, IM, and nasal spray).16,44,45 It is often
emergency medical staff will have no knowledge of the use referenced in zoo and wildlife literature as the emergency
of α-2 specific antagonists and are not experienced in the antagonist of choice for UPO exposure.1–4 Because the
treatment of severe medetomidine overdose. short-acting naloxone does not pharmacologically match
the long duration of action of UPOs, multiple doses may be
Exposure to a Combination of Potent required in clinically significant exposures. Naltrexone is a
Anesthetic Drugs close relative of naloxone but has a significantly longer dura-
tion of action.2,15,16,44,46 It is available for humans in tablet
Accidental human exposure to a combination of drugs may and extended-release injectable suspension forms and is
significantly impact the victim. Combining a UPO with used primarily to manage chronic conditions such as alcohol
midazolam and/or medetomidine may have a multiplicative and opioid dependence. The veterinary formulation is an
effect on depression of cardiorespiratory function. From a injectable solution (50 mg/mL, Naltrexone HCl, Trexonil,
174 SE C T I O N 6 Anesthesia and Analgesia
Wildlife Pharmaceuticals, Inc.) and is the reversal agent of Atipamezole has recently been recommended in case
choice for UPO anesthetized species, because its pharmacol- of an accidental exposure to these potent α-2 agonists in
ogy better matches the long duration of the UPO.2–4 humans20,21 and is further reviewed later in this chapter. See
As mentioned, naloxone is often referenced for use in case Fig. 27.3 for the recommended emergency response to an
of an emergency UPO exposure. Due to the current human exposure of potent α-2 agonists.
opioid overdose dilemma, some state health officials are even
allowing nonmedical professionals in prehospital locations Legal Concerns
to administer naloxone because it has been used to safely
reverse over 10,000 opioid-related overdoses in the United It is beyond the scope of this chapter to review and interpret
States.45,47 Recommendations for the use of naltrexone in the laws pertaining to the actions of veterinarians or emer-
a UPO exposure have been made in the zoo and wildlife gency care personnel responding to a catastrophic human
literature.3,4 These authors suggest providing 25–50 mg IM exposure to a dangerous anesthetic agent used in zoo and
followed by 25–50 mg IV, with the likelihood of naltrexone wildlife. The dilemma for both professions is the legal rami-
side effects being minimal. Because there are currently no fications if we choose to intervene responsibly. Fortunately,
injectable solutions available for humans, the veterinary this type of incident is rare, but it behooves us to understand
formulation would need to be used. It should be noted the moral, ethical, and legal principles of the law which are
that the injectable veterinary formulation of naltrexone in place to protect us in case we are presented with this type
follows the same US Pharmacopeia (USP) reference stan- of unfortunate incident. Two important terms found in
dards enforced by the FDA to ensure the identity, strength, the legal literature are germane when considering the risks
sterility, quality, and purity of human medicines (Bill Lance, involved in intervening in a human medical emergency:
Wildlife Pharmaceuticals, Inc., personal communication). Good Samaritan and Duty to Rescue.
There is no reference in the literature of the use of oral
naltrexone to treat a human exposed to an UPO. This Good Samaritan Law
may be considered only in a coherent, conscious patient This law states that citizens should not be discouraged from
with the ability to swallow a tablet. The slower onset of helping others at a fundamental level commensurate with
action of the oral antagonist would need to be considered. their expertise by fear of liability.51–53 All 50 states and the
A possible scenario might include a minor or questionable District of Columbia in the United States have some type
exposure where the injectable product was not available, of Good Samaritan law, but they vary by jurisdiction who is
refused by the patient, or considered unnecessary given the protected from liability and under what circumstances. This
circumstance. The authors agree that both naltrexone and is especially true if your action is reasonable, without decep-
naloxone should be considered in the case of an emergency tion and commensurate with your training, knowledge,
UPO exposure. See Fig. 27.2 for the resuscitation algorithm and ability. Prior to engaging in a compassionate act, the
for human exposure to a UPO. cautious veterinarian would assess the situation, determine
if he or she would competently be able to provide some
Atipamezole Use in Humans assistance without doing further damage, and then do what
he or she can to help. If the patient is unconscious, delu-
The use of atipamezole in the emergency response to an sional, intoxicated, or in imminent peril, there is implied
accidental exposure to the ultra-potent concentrated form consent if the assistance is reasonable and not negligent. It
of medetomidine used in zoo and wildlife has not been appears doubtful that a court would consider a compas-
thoroughly reviewed until recently. The safety and efficacy sionate veterinarian illegally practicing human medicine for
of atipamezole in humans has been investigated20,21,48,49 reasonably tending to a catastrophic exposure of a drug we
and is discussed further later in this chapter. It should be know is extremely dangerous to humans.
noted that atipamezole is currently not available for use
in humans, internationally. In comparison with veterinary Duty to Rescue
species, the human is more sensitive on a mg/kg basis to the In general, there is no law in the United States that obligates
α-2 agonist effects of dexmedetomidine and presumably to you to aid someone who is in danger, but there are special
its close relative, medetomidine.20,21 Furthermore, humans circumstances that may impose a moral or ethical duty
appear to be less sensitive to the effects of atipamezole when upon you to rescue.51,52 An example may include a situation
used as an α-2 antagonist for dexmedetomidine, and this in which you have a unique relationship and perceived
would require higher doses than in veterinary species. In obligation with the person in danger or if your negligence
domestic and nondomestic animals, atipamezole is recom- or action caused the need for rescuing. Legal definitions
mended at a 5:1 ratio to the mg dose of medetomidine and responsibilities for duty to rescue may vary by state
and 10:1 ratio to dexmedetomidine.6,18,19 It is significant and municipality. If an accident occurs, you may have a
to note that atipamezole is routinely and safely used to duty to rescue at some level due to your leadership role
reverse medetomidine and dexmedetomidine for anesthetic as veterinarian, knowledge of the drugs and their effects,
procedures in the gorilla, chimpanzee, and orangutan—our and ability to provide competent assistance due to medical
closest human relatives.22,24,50 training.
CHAPTER 27 Use of Naltrexone and Atipamezole in Emergency Response to Human Exposure to Ultra-Potent Opioids 175
30. Margetts L, Sawyer R: Transdermal drug delivery: principles and 51. DeGioa P: Human medical emergencies pose conundrum for
opioid therapy, Contin Educ Anaesth Crit Care Pain 7(5):171–176, veterinarians. Many are unsure of their responsibility and liability.
2007. Veterinary Information Network (VIN), 2013. Retrieved from
31. N’Da DD: Prodrug strategies for enhancing the percutaneous http://news.vin.com/vinnews.aspx?articleId=27398.
absorption of drugs, Molecules 19:20780–20807, 2014. Retrieved 52. Rollins BE: An ethicist’s commentary on when a veterinarian can
from www.mdpi.com/journal/molecules. render medical assistance to people, Can Vet J 44(3):189–191,
32. Stanley TH: The fentanyl story, J Pain 15(12):1215–1226, 2014. 2003.
33. Stanley TH, Haue B, Mock DL, et al: Oral transmucosal 53. Sutton V: Is there a doctor (and a lawyer) in the house? Why
fentanyl citrate (lollipop) premedication in human volunteers, our Good Samaritans laws are doing more harm than good for
Anesth Analg 69:21–27, 1989. a national public health security strategy: a fifty-state survey, J
34. Ramsay EC, Sleeman JM, Clyde VL: Immobilization of black Health Biomed L 7:261–300, 2010.
bears (Ursus americanus) with orally administered carfentanil 54. American Heart Association. Basic Life Support (BLS)
citrate, J Wildl Dis 31(3):391–393, 1995. guidelines, 2014. https://eccguidelines.heart.org/index.php/
35. Mama KR, Steffey EP, Withrow SJ: Use of orally administered circulation/cpr-ecc-guidelines-2/part-5-adult-basic-life-support-
carfentanil prior to isoflurane-induced anesthesia in a Kodiak and-cardiopulmonary-resuscitation-quality/.
brown bear, J Am Vet Med Assoc 217:546–549, 2000. 55. Petrini KR, Keyler DE, Ling L, et al: Immobilizing agents –
36. Mortenson J, Bechert U: Carfentanil citrate used as an oral developing an urgent response protocol for human exposure,
anesthetic agent for brown bears (Ursus arctos), J Zoo Wildl Med Proceedings of AAZV Annual Conference, St. Louis, MO, pp
32(2):217–221, 2001. 147–155, 1993.
37. Pollock CG, Ramsay EC: Serial immobilization of a Brazilian 56. Insys Advisory Committee: Naloxone for Treatment
tapir (Tapirus terrestrus) with oral detomidine and oral carfent- of Opioid Overdose. https://www.fda.gov/downloads/
anil, J Zoo Wildl Med 34(4):408–410, 2003. AdvisoryCommittees/CommitteesMeetingMaterials/Drugs/
38. Haymerle A, Fahlman A, Walzer C: Human exposures to immo- AnestheticAndAnalgesicDrugProductsAdvisoryCommittee/
bilising agents: results of an online survey, Vet Rec 167:327–332, UCM522690.pdf.
2010. 57. Webster LR, Johnson FK, Stauffer J, et al: Impact of intravenous
39. Dowell D, Haegerich TM, Chou R: CDC guideline for prescrib- naltrexone on intravenous morphine-induced high, drug liking,
ing opioids for chronic pain - United States, 2016, MMWR Morb and euphoric effects in experienced, nondependent male opioid
Mortal Wkly Rep 65(1):1–50, 2016. users, Drugs R D 11(3):259–275, 2011.
40. Equianalgesic. Opioid conversion chart. Retrieved from https:// 58. Crabtree B: Review of naltrexone, a long-acting opiate antagonist,
en.wikipedia.org/wiki/Equianalgesic. Clin Pharm 3(3):273–280, 1984.
41. Wecker L: Drugs to control pain. In Wecker L, Crespo L, 59. Bharati S, Pal A, Biswas C, et al: Incidence of cardiac arrest
Duanway G, et al, editors: Brody’s human pharmacology, ed 5, increases with the indiscriminate use of dexmedetomidine: A
Philadelphia, PA, 2010, Mosby Elsevier, pp 391–409. case series and review of published case reports, Acta Anaesthesiol
42. Coetzee JF: Safety of pain control with morphine: new (and old) Taiwan 49:165–167, 2011.
aspects of morphine pharmacokinetics and pharmacodynamics, 60. Hoffmann U, Meister C, Golle K, et al: Severe intoxication with
South Afr J Anaesth Analg 16(2):7–15, 2010. the veterinary tranquilizer xylazine in humans, J Anal Toxicol
43. Weese J, Faires M: A survey of needle handling practices 25(4):245–249, 2001.
and needlestick injuries in veterinary technicians, Can Vet J 61. Ingersoll-Weng E, Manecke G, Thistlethwaite P: Dexmedetomi-
50(12):1278–1282, 2009. dine and cardiac arrest, Anesthesiology 100(3):738–739, 2004.
44. Pani N, Dongare PA, Mishra RK: Reversal agents in anaesthesia 62. Anderson R, Hart G, Crumpler C, et al: Clonidine overdose:
and critical care, Indian J Anaesth 59(10):664–669, 2015. report of six cases and review of the literature, Ann Emerg Med
45. Wheeler E, Jones TS, Gilbert MK, et al: Opioid overdose 10(2):107–112, 1981.
prevention programs providing naloxone to laypersons - United 63. Osterhoudt KC: Clonidine and related imidazoline poisoning.
States, 2014, MMWR Morb Mortal Wkly Rep 64(23):631–635, Up to Date. https://www.uptodate.com/contents/clonidine-and
2015. -related-imidazoline-poisoning.
46. Gonzalez J, Brogden R: Naltrexone. A review of its pharmacody- 64. Roberge R, Mcguire S, Krenzelok E: Yohimbine as an antidote
namic and pharmacokinetic properties and therapeutic efficacy for clonidine overdose, Am J Emerg Med 14(7):678–680, 1996.
in the management of opioid dependence, Drugs 35(3):192–213, 65. Schieber R, Kaufman N: Use of tolazoline in massive clonidine
1988. poisoning, Am J Dis Child 135(1):77–78, 1981.
47. Centers for Disease Control and Prevention: Opioid Overdose. 66. Karhuvaara S, Kallio A, Scheinin M, et al: Pharmacological
Retrieved from https://www.cdc.gov/drugoverdose/. effects and pharmacokinetics of atipamezole, a novel alpha-2
48. Huupponen R, Karhuvaara S, Anttila M, et al: Buccal delivery adrenoreceptor antagonist – a randomized, double blind cross
of an α2-adrenergic receptor antagonist, atipamezole, in humans, over study in healthy male volunteers, Br J Clin Pharmacol
Clin Pharmacol Ther 58(5):506–511, 1995. 30:97–106, 1990.
49. Scheinin H, Aantaa R, Anttila M, et al: Reversal of the sedative 67. Karhuvaara S, Kallio A, Salonen M, et al: Rapid reversal of
and sympatholytic effects of dexmedetomidine with a specific alpha 2-adrenoceptor agonist effects by atipamezole in human
alpha 2-adrenoceptor antagonist atipamezole: a pharmacody- volunteers, Br J Clin Pharmacol 31(2):160–165, 1991.
namic and kinetic study in healthy volunteers, Anesthesiology 68. Link MS, Berkow LC, Kudenchuk PJ, et al: Part 7: Adult
89(3):574–584, 1998. Advanced Cardiovascular Life Support: 2015 American Heart
50. Adami C, Wenker C, Hoby S, et al: Evaluation of effectiveness, Association Guidelines Update for Cardiopulmonary Resuscita-
safety and reliability of intramuscular medetomidine-ketamine tion and Emergency Cardiovascular Care, Circulation 132(18
for captive great apes, Vet Rec 171(8):196, 2012. Suppl 2):S444–S464, 2015.
28
Vaporizers and Field Anesthesia
Equipment for Free-Ranging Wildlife
SATHYA K. CHINNADURAI
Introduction drugs are readily expelled from the body during the course
of recovery, there is minimal potential for prolonged drug
Performing general anesthesia in the field may be technically residues.11 Disadvantages include the need to transport
challenging when equipment designed for a single location volatile fluids, expense and bulk of the vaporizers, and
(e.g., hospital use) must be modified for use in remote logistical concerns of transporting compressed gases.12,13 In
locations with limited access to electrical power and oxygen. some cases, inhalant anesthesia may be chosen to avoid
There is the additional challenge of having sufficient sup- having to use tightly regulated controlled substances; this is
plies transported to and stored at a field site.1 Capture- and especially true crossing international boundaries.
anesthesia-related morbidity and mortality may occur with
field immobilizations, regardless of anesthetic protocols.2,3 Basic Inhalant Pharmacology
In general, the two broad categories of anesthetic agents are
inhaled or injectable agents. Because field anesthesia needs A set of basic definitions necessary for understanding inhal-
to be simple, safe, and easily mobile, injectable anesthesia ant anesthetic and vaporizer use is provided in Box 28.1.
is often used instead of inhalant anesthesia for logistical With the exception of nitrous oxide, all commonly used
reasons.1 Use of volatile inhalant anesthesia in the field inhalant anesthetics are a vapor at room temperature and
is a tradeoff between the rapid induction and recovery not truly a gas. A vapor is the gaseous state of a substance
associated with inhalants and the logistical difficulty of that is a liquid at ambient temperature and pressure. A
moving compressed gas cylinders and vaporizers into the gas may be delivered at a concentration between 0% and
field. Injectable only protocols lack some of the ability 100%, whereas a vapor has a maximum concentration that
for “fine-tuning” by means of incremental adjustments in is determined by its vapor pressure.10 The amount of an
depth. In addition, there is the possibility of prolonged inhalant anesthetic in a mixture may be expressed as a
drug effect and renarcotization. This chapter focuses on the volume percent or a partial pressure. When administering
use of inhalant anesthesia, with descriptions of the types of a vaporized inhalant anesthetic, the end goal is to achieve a
vaporizers and novel delivery systems and ventilators that partial pressure of anesthetic in the brain and spinal cord that
may be adapted to the challenges of the field setting. results in anesthesia.14 A series of concentration gradients
needs to be established to allow movement of anesthetic
Inhalant Anesthesia from the vaporizer, through the circuit to the lung, and then
from the alveoli to the blood and then the central nervous
Inhalant anesthetics are widely used in a clinical setting system. Multiple factors may affect the rate of equilibration
and possess unique advantages and disadvantages for use between the vaporizer and the brain, including vaporizer
in a field setting. With careful attention to the principles setting, fresh gas flow rate, inspired anesthetic concentra-
of inhalant anesthetic pharmacology and appropriate anes- tion, circuit volume, alveolar ventilation, cardiac output,
thetic equipment use, wildlife veterinarians have successfully and solubility of the inhalant.10,14
used inhalants in a wide variety of species in a multitude of The commercially available inhalant anesthetics differ in
natural settings. Use of inhalants for avian, marine mammal, terms of potency, solubility, and vapor pressure. Isoflurane
and small rodent anesthesia is well described in the wildlife and sevoflurane are the two most commonly used inhal-
medicine literature.4–9 Unlike injectable anesthetics, inhal- ant agents in field veterinary medicine. Isoflurane has a
ants are administered and eliminated via the respiratory higher blood solubility compared with sevoflurane, owing
system. This allows for a rapid and precise adjustment of to a higher blood to gas partition coefficient. This higher
the anesthetic depth of the patient.10 Because many of these solubility translates to a slower induction and recovery in
177
178 SE C T I O N 6 Anesthesia and Analgesia
most species. Isoflurane is also more potent than sevoflu- Table 28.1. A volatile anesthetic allowed to vaporize in a
rane, meaning that a lower partial pressure of isoflurane in closed container (i.e., the vaporizer) will reach a concentra-
the brain is necessary for anesthesia. Potency is typically tion proportional to its saturated vapor pressure. For example,
described in terms of minimum alveolar concentration isoflurane has a saturated vapor pressure of 240 mm Hg.
(MAC; see Box 28.1). The commonly used inhalation So, if liquid isoflurane is allowed to vaporize at atmospheric
anesthetics in veterinary medicine are described in pressure (760 mm Hg) in a closed container, the saturated
vapor concentration of isoflurane in the container will be
240/760 × 100, or 32%. In contrast, sevoflurane has a
• BOX 28.1 Terminology, Abbreviations, and
saturated vapor pressure of 160 mm Hg and thus reaches
Units of Measure Used to Describe a maximum concentration of 21% (160/760 × 100 = 21).
Vaporizers and Inhalant Anesthetics This is important to understand for open drop anesthesia
and if a vaporizer is tipped, as described later.
Vapor pressure: Partial pressure of a vapor over the liquid. Vapor Most anesthetics have a saturated vapor concentration
pressure is dependent on temperature; at higher temperatures, well above therapeutic levels and, if they were allowed to
a liquid will have a higher vapor pressure.
Saturated vapor pressure: The maximum vapor pressure in a
vaporize freely and be breathed by the patient, could be
closed container at equilibrium. At this point, for every molecule lethal. For this reason the preferred way of delivering inhal-
that enters the gaseous phase from the liquid, one molecule ant gas anesthetic is by using a precision, agent-specific
enters the liquid phase. Like vapor pressure, saturated vapor vaporizer and compressed oxygen.
pressure is temperature dependent.
Delivered anesthetic concentration: The concentration of
the anesthetic vapor in a mixture of gases. Concentration is Vaporizers
expressed as a volume percentage (%). The concentration is
dependent on the partial pressure of the vapor or gas being Most commonly used inhalant anesthetics are delivered by
described and the other gases in the mixture. a vaporizer with some fresh gas source. The only common
Blood to gas partition coefficient: A partition coefficient is exception is nitrous oxide, which is regulated by flowmeter
the ratio of the concentration of a gas in two separate media at
equilibrium and is a measure of solubility in dissimilar fluids. The
alone. Most modern vaporizers are variable bypass, concen-
blood to gas partition coefficient for an inhalant is the ratio of tration calibrated, and temperature and flow compensated.
the concentration of the inhalant in the blood versus in the gas These features are designed to reduce the chance of user
phase. Drugs with a higher blood to gas coefficient are more error. Nonprecision (uncalibrated) vaporizers are rarely in
soluble in blood and thus have slower induction and recovery use in veterinary medicine but are, on occasion, used in
times.
Potency: Potency of inhalant anesthetics is typically defined
field settings.15 As mentioned previously, delivered amounts
in terms of MAC. The term “minimum anesthetic concentration” of inhalant anesthetics may be quantified either by partial
is used for those species without alveoli. MAC may be used to pressure or concentration in volume percentage. Most clini-
compare anesthetics with one another. cians are more familiar with delivered concentrations as
Latent heat of vaporization: Calories needed to change 1 g determined by the percentage setting on the vaporizer. For
of liquid to vapor. As a liquid vaporizes, the remaining liquid will
cool due to energy lost in the vaporization process. This is the
example, a precision isoflurane vaporizer set to 3% should
reason that an anesthetic liquid will cool during vaporization. deliver 3% isoflurane in 97% carrier gas (typically oxygen).
Specific heat: Heat needed to raise 1 g of material To understand the function and use of vaporizers in the
1°C. Copper in a vaporizer has a high specific heat, so the field, it is essential to understand a certain set of terminology
temperature change is minimal during vaporization. (see Box 28.1). Because the saturated vapor concentration
Units of measure for pressure and their conversion: 100 kPa
= 1000 mbar = 1 bar = 760 mm Hg = 1030 cm H2O = 14.7 psi
of an anesthetic is always well above the clinically useful
= 1 atm. volume percentage used, it is necessary to dilute the inhal-
ant vapor with fresh gas (oxygen, a combination of oxygen
MAC, Minimum alveolar concentration.
and nitrous oxide, or room air). For example, the saturated
vapor concentration of isoflurane at 20°C and 1 atm is
TABLE
28.1 Selected Properties of Commonly Used Inhalant Anesthetics
Vapor Pressure (mm Hg) Vapor Pressure (mm Hg) Blood to Gas
Anesthetic @ 20°C @ 24°C Partition Coefficient MAC (in Dogs, %)
Isoflurane 240 286 1.4 1.3
Sevoflurane 160 183 0.68 2.36
Desflurane 700 804 0.42 7.2–9
Methoxyflurane 23 28 12 0.29
CHAPTER 28 Vaporizers and Field Anesthesia Equipment for Free-Ranging Wildlife 179
32%, which is likely lethal to all vertebrate species. The available for laboratory animal use and has been modified
vaporizer functions to dilute the 32% isoflurane vapor with for field use.17
oxygen to achieve a clinically useful percentage (0%–5%). Inhalant anesthetics delivered by an “open drop” method,
Most vaporizers in clinical use for both veterinary and in which the anesthetic is applied to a cotton ball or gauze
human medicine are precision, out-of-circuit vaporizers. and allowed to spontaneously vaporize in a closed container,
These vaporizers are agent-specific, concentration-calibrated have been used extensively in rodent anesthesia.18 As men-
machines that provide a regulated volume percentage of tioned previously, anesthetics allowed to vaporize in this
inhalant anesthetic.16 Typically, they function by a vari- fashion will reach a concentration dictated by their vapor
able bypass mechanism that allows a certain portion of the pressure. Isoflurane will reach a maximum concentration of
carrier gas (typically oxygen) to flow over a pool of liquid 32%, whereas sevoflurane will reach a maximum concen-
anesthetic until it vaporizes to saturation. The remainder tration of 21%. Open drop delivery results in extremely
of the carrier gas passes through a bypass chamber, and high concentrations of volatile anesthetic, in many cases
the two paths are mixed to achieve the desired concentra- far exceeding lethal doses. This method should be used
tion set on the vaporizer dial. If a vaporizer is tipped over only by experienced personnel, with a previously measured
(typically greater than 45 degrees), liquid anesthetic will container volume and calculated amounts of anesthetic. In
flood the bypass chamber and the vaporizer will release an addition, the open drop method should be used only for
incredibly high concentration of anesthetic, at or near its brief induction, not continued maintenance, of anesthesia
saturated vapor concentration (e.g., 32% for isoflurane). and reserved for situations when transport of the compressed
Most modern vaporizers are temperature, flow, and back oxygen and a vaporizer are impossible.1
pressure compensated, although these factors are rarely
considered in controlled hospital use. These compensation Gas Anesthesia at Altitude
mechanisms have limits, and it is important to remember
that the vaporizers are only temperature compensated for Changes in barometric pressure may affect the output of a
a range of 15°C–35°C such that the amount of vapor- precision vaporizer. This issue is rarely encountered in the
ized anesthetic will be decreased at colder temperatures hospital setting but may be common in field settings. This
and increased at higher temperatures.16 Similarly, regarding is due to the changing ratio of ambient pressure and anes-
flow compensation, most modern vaporizers are accurate thetic vapor pressure that depends on altitude. Vaporizers
between 0.25 and 15 L/min of fresh gas flow.12 are calibrated at 20°C at sea level (1 atm or 760 mm Hg
One notable exception to the typical vaporizer design is barometric pressure). At a higher altitude, ambient pressure
the desflurane vaporizer. Desflurane is highly volatile and is less than 1 atm, but the vapor pressure (in mm Hg) of the
boils at room temperature. Due to its high latent heat of anesthetic does not change; thus delivered concentration
vaporization, desflurane rapidly cools during vaporization, for a given vaporizer setting will increase.10 For example, at
and this would overwhelm the insulating capacity of a 760 mm Hg, the saturated vapor concentration of sevoflu-
regular vaporizer, so the desflurane vaporizer needs to be rane is 21% (equation earlier). If the same drug vaporizes
thermostatically controlled to keep it at 39°C. In addition, at an ambient pressure of 632 mm Hg (5000 ft elevation),
because desflurane vaporizes so extensively, it would need an the saturated vapor concentration is 25% (160/632 × 100
infeasibly high fresh gas flow rate in a traditional vaporizer. = 25). Thus a sevoflurane vaporizer set at 5% will deliver
Instead, no fresh gas goes into the desflurane sump, and it a slightly higher concentration when used at 632 mm Hg
releases pure desflurane vapor into the mixture, which is than at 760 mm Hg.
then diluted with oxygen.12 The need for electrical power This is made more confusing by the fact that the effect of
limits the use of desflurane in the field. the anesthetic is still determined by potency, as expressed in
Nonprecision, in-circuit vaporizers are no longer in terms of MAC. MAC as a partial pressure does not change
common use in veterinary medicine, but they do still with altitude, but MAC expressed as a volume percentage
occasionally find a place in field anesthesia. The units are does change. At higher altitude, the vaporizer will put out
small and light and have low resistance to breathing.15 These a higher volume percentage but the same partial pressure.
devices may be used with low-potency anesthetics with a So, although the delivered percentage might be higher, the
low vapor pressure. It should be noted that the delivered animal’s MAC expressed as a volume percentage increases
percentage cannot be precisely controlled with a dial, so use proportionally and the effect of a given vaporizer setting
with currently available inhalant anesthetics may lead to a will be similar at different elevations.10 Changes in ambient
fatal overdose. The delivered concentration of anesthetic gas pressure may also affect the accuracy of a flowmeter that is
may vary with patient ventilation and fresh gas flow rate; calibrated at sea level.
thus the potential for user error is high. The only way to
measure delivered inhalant concentrations is by using a gas Anesthesia Machines
analyzer, which is rarely feasible in a field setting. Another,
less traditional, mechanical vaporizer uses a syringe of liquid The vaporizer is just one component needed for volatile anes-
anesthetic delivered in a precise amount to achieve a desired thetic administration. The anesthetic circuit requires oxygen
concentration when mixed with pumped ambient air. It is or another compressed carrier gas, a pressure regulator with
180 SE C T I O N 6 Anesthesia and Analgesia
in Figs. 28.1 and 28.2. Components Labeled. The machine may be set up as a rebreather
In many cases, due to logistical ease, nonrebreathing (A) and a nonrebreather (B). The components of the system include (1)
systems are used for field anesthesia machines. It should be oxygen tank and regulator, (2) vaporizer, (3) carbon dioxide adsorbent
canister (rebreathing system only), (4) pressure manometer (nonre-
noted that a nonrebreathing system is not physiologically breathing system only), (5) adjustable pressure limiting (pop-off) valve,
appropriate for animals larger than 30 kg and ideally is (6) rebreathing bag, and (7) scavenge hosing.
not used on animals greater than 10 kg. The perceived
advantage of a nonrebreathing system is the simpler design,
with fewer parts and less potential for mechanical failure. would require an oxygen flow rate of 3 L/min, whereas
Unfortunately, a nonrebreathing system also requires a a circle system would require 300 mL/min (Box 28.2). If
much higher oxygen flow rate and expends more volatile that flow rate is not met, it is highly likely that the patient
anesthetic agent, making it much less suitable for field use. will inhale its own exhaled gases, including carbon dioxide.
Maintaining a 10-kg animal on a nonrebreathing system When using a nonrebreathing system on a larger animal in
CHAPTER 28 Vaporizers and Field Anesthesia Equipment for Free-Ranging Wildlife 181
Oxygen Safety
Transporting oxygen into the field is not without hazard.
Because the cylinders are under high pressure, it is pos-
sible for them to rupture, if not handled appropriately,
and should be handled only by trained personnel. Unsafe
practices include storing cylinders upright, instead of on
their side, transporting them for distances by hand without
a proper cart, and exposure to extreme temperatures.
Unfortunately, many of these practices are commonplace
and occasionally necessary in a field setting but should
be minimized whenever possible. Exposure to extreme
temperatures should be avoided at all costs; temperatures
greater than 54°C (130°F) and less than −7°C (20°F) may
• Figure 28.3 Oxygen pressure regulator with a pressure gauge damage an oxygen tank, making them dangerous to use.12
and an integrated flowmeter that may be used to supply a portable Even more modest fluctuations in temperature may affect
anesthesia machine through the attached hose or may be connected the pressure of the gas inside a fixed volume canister. Most
to an oxygen demand valve.
tanks have a pressure relief mechanism built into the valve.
This pressure relief system will allow the contents of the
TABLE Oxygen Cylinder Size, Volume, and Pressure tank to vent rapidly before the tank itself would explode due
28.2 When Filled at 21°C to over pressure. Dropping a tank could result in damage
Volume When Pressure When to the tank or release of the pressure relief mechanism and
Cylinder Size Full Full rapid discharge of the contents, which may turn the tank
or any surrounding loose material into a deadly projectile.
B 200 1900 Oxygen is not flammable by itself but is an oxidizing
D 400 1900 agent, and, if a flammable material and a source of ignition
E 660 1900
(flame or spark) are present, a fire may occur. Fires in an
enriched oxygen environment will burn hotter and faster,
M 3450 2200 and, if the fire involves oxygen under pressure, an explosion
H 6900 2200 may occur. Transfilling, or filling a smaller oxygen cylinder
(E tank) from a larger one (H tank), may be common
practice in a field setting. Rapidly filling an empty small
is critical in a field setting where replacement oxygen tanks container from a large one at high pressure will cause the
and filling sources may be days away. Tank pressure is read smaller cylinder to heat up rapidly due to recompression
out on a gauge on the pressure regulator. Because oxygen of the gas, and the resulting heat could ignite nearby flam-
is a gas, the volume in the tank is directly proportional to mable materials.12
the pressure in the tank, assuming that the tank is kept at a Multiple safety mechanisms are used to prevent acci-
constant temperature. So a tank that was filled to 660 L at dental mixing of compressed gases; the diameter index
1900 psi will contain 330 L when the tank pressure reads safety system (DISS) uses a standard diameter and thread
950 psi and 165 L when the pressure is 475 psi.12,16 Table configuration to prevent oxygen lines from being connected
28.2 lists common cylinder volumes and pressures. Oxygen to fittings for other gases and vacuum. Similarly, the pin
cylinders may be made of steel alloy, which is stronger, or index safety system prevents oxygen tank valves from being
aluminum, which is lighter. connected to regulators for other gases. It is imperative that
In the planning stages of a field project, it is critical all connections and fittings be tested before transporting
to understand how much working time is possible with a equipment to a remote setting, because appropriate connec-
given number of tanks. Similarly, efficient use of oxygen in tions and fittings may not be easily available.
the field is critical to avoid running out. For example, the Portable oxygen concentrators are battery-powered,
following equations may be used to determine how much compact, portable devices that may be used in the field for
oxygen is needed if the plan is to anesthetize three 50-kg supplemental oxygen. They may be more convenient and
animals for 1 hour each. On a circle system, the ideal fresh portable than oxygen cylinders. These units use room air
gas flow rate from Box 28.2 would be 1.5 L/min (30 mL/ and, by extracting the nitrogen, may dispense 90%–96%
min × 50 kg = 1500 mL/min = 1.5 L/min). oxygen.21
3 animals × 1 h animal × 60 min h × 1.5 L min
= 270 L of oxygen. Field Ventilatory Support
The 270 L of oxygen is less than half the volume of a full Multiple devices may provide positive pressure ventilation in
E tank, and so this could be easily accomplished. remote locations. Bag-valve devices, also known as manual
CHAPTER 28 Vaporizers and Field Anesthesia Equipment for Free-Ranging Wildlife 183
flavescens) being ventilated with a bag-valve mask and monitored with • Figure 28.6 A modified leaf blower used to ventilate an anesthe-
battery-powered pulse oximetry and capnography. (Photo courtesy of tized African elephant (Loxodonta africana). (Photo courtesy of the
the Chicago Zoological Society.) North Carolina Zoo.)
mammals or large ungulates, these data may prove essential 7. Heath RB, Delong R, Jameson V, et al: Isoflurane anesthesia in
in assessing the patient under anesthesia before a crisis or free ranging sea lion pups, J Wildl Dis 33(2):206–210, 1997.
cardiopulmonary arrest occurs. Such information may also 8. Gales NJ, Mattlin RH: Fast, safe, field-portable gas anesthesia for
otariids, Mar Mammal Sci 14(2):355–361, 1998.
prove useful in assessing instances of postrelease mortality
9. Kocer CJ, Powell LAA: Field system for isoflurane anesthesia of
of treated animals. Lactate measurements provide invalu- multiple species of mesopredators, Am Midl Nat. 161:406–412,
able information about adequacy of perfusion and oxygen 2009.
delivery and may be used as an indicator of exertional 10. Steffey E, Mama K, Brosnan R: Inhalation anesthetics. In Grimm
myopathy. K, Lamont L, Tranquilli W, et al, editors: Lumb and Jones’ veteri-
nary anesthesia, ed 5, 2015, Wiley-Blackwell, pp 297–331.
Conclusion 11. Papich MG: Drug residue considerations for anesthetics and
adjunctive drugs in food-producing animals, Vet Clin North Am
Inhalant anesthetics may be safely and effectively used in Food Anim Pract 12(3):693–706, 1996.
field settings on a wide variety of species. A basic understand- 12. Dorsch J, Dorsch S: Understanding Anesthesia Equipment, ed
ing of inhalant pharmacology and vaporizer and anesthetic 5, Philadelphia, 2008, Wolters Kluwer, Lippincott Williams &
Wilkins.
circuit design and function is needed to use this equip-
13. International Air Transport Association: IATA dangerous goods
ment safely. Each type of machine has inherent benefits regulations, ed 54, Montreal, Canada, 2013, International Air
and limitations that determine its appropriateness for the Transport Association.
species of interest. In most situations, wildlife veterinarians 14. Lamont L, Grimm K: Clinical pharmacology. In West G, Heard
and anesthetists will need to modify commercially available D, Caulket TN, editors: Image result for zoo and wild animal
equipment or purpose-build units to meet the needs of the immobilization and anesthesia zoo animal and wildlife immobiliza-
species of interest in their natural habitat. tion and anesthesia, ed 2, Ames, IA, 2014, John Wiley & Sons,
Inc., pp 5–12.
15. Lewis JCM: Field use of isoflurane and air anesthetic equipment
Acknowledgments in wildlife, J Zoo Wildl Med 35(353):303–311, 2004.
Sections of this chapter were previously published in 16. Mosley C: Anesthesia equipment. In Grimm K, Lamont L,
Tranquill IW, et al, editors: Lumb and Jones’ veterinary anesthesia,
Chinnadurai SK, Strahl-Heldreth D, Fiorello CV, Harms
ed 5, Amea, IA, 2015, Wiley-Blackwell, pp 23–84.
CA: Best-practice guidelines for field-based surgery and 17. Gorini SJ, Wedul JM, Arnemo JDC, et al: Field anesthesia of
anesthesia of free-ranging wildlife. I. Anesthesia and anal- least weasels (Mustela nivalis nivalis) with isoflurane, Wildl Biol
gesia. J Wildl Dis 52(2s):S14–S27, 2016 and are used with Pr 1:7–13, 2013.
permission of the journal. 18. Parker WT, Muller LI, Gerhardt RR, et al: Field use of isoflu-
rane for safe squirrel and woodrat anesthesia, J Wildl Manage
72(5):1262–1266, 2008.
References 19. American College of Veterinary Anesthesia and Analgesia: Com-
mentary and recommendations on control of waste anesthetic
1. Chinnadurai SK, Strahl-Heldreth D, Fiorello CV, et al: Best- gases in the workplace. [www.acvaa.org], 2013.
practice guidelines for field-based surgery and anesthesia of 20. Smith JC, Bolon B: Comparison of three commercially avail-
free-ranging wildlife. I. Anesthesia and analgesia, J Wildl Dis able activated charcoal canisters for passive scavenging of waste
52(2s):S14–S27, 2016. isoflurane during conventional rodent anesthesia, J Am Assoc Lab
2. Arnemo JM, Ahlqvist P, Andersen R, et al: Risk of capture- Anim Sci 42(2):2003.
related mortality in large free-ranging mammals: experiences 21. Fahlman Å, Caulkett N, Arnemo J, et al: Efficacy of a portable
from Scandinavia, Wildlife Biol 12(1):109–113, 2006. oxygen concentrator with pulsed delivery for treatment of hypox-
3. DelGiudice GD, Sampson BA, Kuehn DW, et al: Understanding emia during anesthesia of wildlife, J Zoo Wildl Med 43(1):67–76,
margins of safe capture, chemical immobilization, and handling 2012.
of free-ranging white-tailed deer, Wildl Soc Bull 33(2):677–687, 22. Jeon M, Mama KR, Zuba JR, et al: Evaluation of blood gas
2005. values in anesthetized southern white rhinoceros (Ceratotherium
4. Desmarchelier M, Cheveau M, Imbeau L, et al: Field use of simum) ventilated with a novel demand ventilator in a zoological
isoflurane as an inhalant anesthetic in the American marten park setting. J Zoo Wildl Med 48(4):1016–1025, 2012.
(Martes americana), J Wildl Dis 43(4):719–725, 2007. 23. Citino SB, Bush M, Rivera O: Simple, unique field ventilator for
5. Small MF, Baccus JT, Waggerman GL: Mobile anesthesia unit large ungulates: another use for your leaf blower. In Proceedings
for implanting radiotransmitters in birds in the field, Southwest AAZV, AAWV, AZA/NAG Joint Conference 2007:51–52.
Nat 49(2):279–282, 2004. 24. Horne WA, Tchamba MN, Loomis MR: A simple method of
6. Machin KL, Caulkett NA: Evaluation of isoflurane and propofol providing intermittent positive-pressure ventilation to etorphine-
anesthesia for intraabdominal transmitter placement in nesting immobilized elephants (Loxodonta africana) in the field, J Zoo
female canvasback ducks, J Wildl Dis 36(2):324–334, 2000. Wildl Med 32(4):519–522, 2001.
29
Perianesthetic Monitoring: Equipment
and Interpretation
KHURSHEED MAMA
“For every mistake that is made for not knowing, a hundred are made
for not looking.” access. It may also be disruptive to the surgeon during the
procedure.
Anonymous
Palpation of a Pulse
Although straightforward, pulse palpation is not always pos-
Introduction sible, either as a result of extreme vasoconstriction due to
large doses of certain medications (e.g., alpha-2 adrenergic
Most anesthetic and adjuvant drugs compromise patient agonists such as medetomidine) or lack of easy access to
homeostasis. Further compromise may result from the externally palpable arteries (e.g., snake).
animal’s physical condition, disease processes, and planned
medical or surgical procedure. Untoward events may occur Use of Pulse Monitors (e.g., Doppler, Pulse
suddenly and, in the absence of intervention, have disastrous Oximeter, Arterial Pressure Waveform)
consequences. The degree of animal monitoring should be Although these tools too are not uniformly applicable
risk-based and inform about physiologic changes. This and may not provide accurate values for blood pressure
chapter reviews available monitoring tools and discusses or saturation in all species, they can inform on heart rate.
their usefulness and limitations. Interpretation and integra- For example, the Doppler may be placed over the heart in
tion of information provided by the selected equipment a reptile and provide an audible signal. Both the Doppler
in conjunction with observation of the animal facilitate and pulse oximeter are available in compact and battery-
appropriate intervention. powered units.
Electrocardiogram
Monitoring the Cardiovascular System
The rate is derived from a tachometer and displayed
Normal cardiovascular function is essential for the mainte- digitally—the tachometer should record a beat with every
nance of adequate oxygen delivery to the tissues. Oxygen QRS complex. On occasion, depending on the amplitude
delivery = cardiac output (CO) × O2 content. CO is deter- of the other depolarization waves, it may count them or
mined by two intrinsic factors (heart rate and myocardial other artifacts caused by motion as well, yielding an inac-
contractility) and two extrinsic factors (preload and after- curate value.
load) that functionally couple the heart and vasculature. The first three methods allow for assessment of mechani-
Although CO is occasionally measured in clinical veterinary cal activity of the heart (or circulation) and, in the case of
patients, parameters such as heart rate and rhythm, arterial the Doppler or arterial pressure monitor, may provide some
blood pressure, central venous pressure (CVP), mucous quantification of this. Ranges for expected heart rates are
membrane color, and capillary refill time are often used unlikely to be available for all species and circumstances
to estimate this and thus the adequacy of tissue perfusion. and are further influenced by anesthesia medications. For
example, the alpha-2 agonist drugs (e.g., medetomidine) are
known to cause bradycardia secondary to centrally mediated
Monitoring Heart Rate
sedation and hypertension resulting from vasoconstriction.1–3
Auscultation Using an External or Opioid agonists are reported to have more variable effects.
Esophageal Stethoscope In canine species and nonhuman primates, they may cause
This is feasible in many species but not practical in others, bradycardia and bradyarrhythmias, whereas in equine and
either due to size considerations or inability to gain megavertebrate species, they may cause tachycardia.4–6
185
186 SE C T I O N 6 Anesthesia and Analgesia
heart, 0.73 mm Hg should be subtracted from or added to pseudoaneurysm, and fistula formation are also reported,
the recorded value (1.36 cm water = 1 mm Hg). especially following long-term catheterization. Inappropri-
As a general rule the Doppler may be used on a wide ate use may result in misinterpretation of data and air or
range of animals (to provide an audible pulse signal), thrombus embolization.
whereas oscillometric technology appears to work best Anesthesia generally lowers blood pressure values, and
on patients with regular heart rhythms and a heart rate to ensure perfusion of vital organs, maintenance of a MAP
within the stated “normal” range. Techniques utilizing a of no less than 60 mm Hg or a systolic arterial pressure
cuff may be inaccurate in patients with irregularly shaped (SAP) of greater than 90 mm Hg is recommended in small
or unusually muscular extremities, in patients with poor animals. It is recommended that the MAP during equine
tissue compliance (e.g., reptiles), etc. Much work has been anesthesia be maintained at least in the range of 70–80 mm
done correlating noninvasive and invasive methods in Hg and that the SAP remain at a value above 100 mm
domesticated and nondomesticated species.20,22–26 Hg to ensure adequate muscle (and organ) perfusion.
Complications reported with use of noninvasive blood Indirect and direct arterial blood pressure values are avail-
pressure monitoring in people include pain, venous stasis, able for other domesticated and nondomesticated species;
compartment syndrome, peripheral neuropathy, and the latter are often obtained during anesthesia, where the
petechiae/ecchymosis. influence of drugs may confound the interpretation of
“normal.”6,8,27–29
Direct Methods Hypotension is common under general anesthesia and
An aneroid manometer or a strain gauge/transducer requires frequently warrants intervention. An intravenous crystalloid
cannulation of an artery and connection of the catheter or colloid fluid bolus and decreasing the dose of inhalation
to a “detector,” using tubing. To maintain accuracy, the anesthetic often resolves the problem. Inotropes and vaso-
tubing should be relatively short and noncompliant. When pressors are used when the former do not lead to resolution
using a strain gauge with a physiologic monitor, a waveform in a timely manner and have dose-dependent and species-
summating sine waves is produced. specific actions. For example, dobutamine is the preferred
To ensure accuracy of the readings, it is also essential that inotrope in horses during inhalation anesthesia and is typi-
the transducer be appropriately balanced (zeroed relative to cally effective at low doses (0.5–2 µg/kg/min).30 Conversely,
atmospheric pressure). The zero reference level is based on higher doses (5–10 µg/kg/min) are needed to improve CO
an estimate of the location of the left ventricular outflow in anesthetized dogs, but a change in blood pressure may
tract (or alternatively the right atrium) and should be main- not be observed. Dopamine (5–7.5 µg/kg/min) will increase
tained for the duration of blood pressure measurement. This blood pressure in dogs and cats; in horses, tachycardia or
is considered the point of the shoulder (or thoracic inlet) for tachydysrhythmias may be observed.31,32 Rabbits, which
patients in dorsal recumbency and midline for patients in often exhibit hypotension during inhalation anesthesia,
lateral recumbency. With single use of modern transducers, show no change even with high doses of dopamine and only
calibration may not be necessary; but with repeated use or minimal improvement after administration of the vasopressor
in the research environment, calibration against a standard phenylephrine.33 These examples highlight the importance
(e.g., mercury or water manometer) is recommended. This of careful monitoring when using these vasoactive drugs,
is especially important when working out of the range of especially in species where basic knowledge of their response
blood pressures of commonly anesthetized domesticated is not known. An alternative that is useful especially when
species and people (e.g., giraffe or elephant where hyperten- ionized calcium values are low is the titrated administration
sion is anticipated). Care should also be taken to adjust of calcium.
the volume of flush and heparin concentration so as to Hypertension is not commonly observed during inhalation
minimize volume overload or heparinization. anesthesia with the exception of adult cattle. Conversely,
A strain gauge provides systolic, diastolic, and mean during injectable anesthesia, most notably when alpha-2
pressure values, whereas the aneroid manometer provides adrenergic agonist drugs are used, blood pressure is often
mean arterial blood pressure values. In field conditions, the elevated. Hypertension to varying degrees is observed in
latter provides a functional and inexpensive way to assess animals with disease (e.g., renal, adrenal) and may neces-
blood pressure directly without the need for electricity and sitate management in the perianesthetic period. Although
expensive equipment. When using the aneroid manometer, the range of normal blood pressure values is not known for
it is the interface between the fluid-filled portion of the all exotic species, hypertension is likely to be more common
line connected to the arterial catheter and the air in the with the use of injectable sedative and anesthesia drugs,
line and not the manometer itself that serves as the zero because many cause vasoconstriction and sympathetic stimu-
reference point. lation. Hence, interpretation of values (if deemed accurate)
Peripheral sites for catheter placement vary with species must be made in light of medications and the species prior
and include the dorsal pedal, digital, auricular, tail, medial, to any intervention. A simple treatment for hypertension
or lateral saphenous artery. Complications of arterial in animals maintained on inhalation anesthetic agents is to
catheterization, while low, include infection, ischemia, and increase the dose and see if blood pressure decreases. If pain
hemorrhage. In human patients, peripheral neuropathy, is considered the cause, analgesic medications should be
188 SE C T I O N 6 Anesthesia and Analgesia
provided. Alternatively, vasodilating drugs or sympatholytic differences (e.g., three- vs. four-chambered hearts) will also
drugs may be used. As an example, acepromazine has been influence “normal” values in nonmammalian species.
used to counter the hypertension caused by ocular admin- The PaCO2 is directly proportional to the CO2 produced
istration of phenylephrine in dogs.34 Vasodilators such as and inversely related to alveolar ventilation. During anes-
hydralazine may also be used if vasoconstriction is suspected thesia, CO2 production should remain relatively consistent
as the cause of hypertension. For sympathetically mediated if body temperature does not vary. Alveolar ventilation, on
hypertension, beta blockers such as esmolol or propranolol the other hand, may vary significantly, as it is influenced
provide a more specific alternative. by the depressant effects of anesthetic drugs, positioning,
and so on. Abdominal distention, surgical manipulation,
Monitoring Central Venous Pressure brain or spinal cord disease, severe metabolic illness, hypo-
thermia, airway obstruction, and other factors may further
CVP is the pressure measured in the thoracic vena cava and influence ventilation in both the awake and anesthetized
used as an indicator of adequate preload in patients with states.
normal myocardial function. It is determined by a complex
interaction of the pumping action of the right heart, blood Blood Gas Analysis
volume, and vascular tone. It may be measured as described The PaCO2 may be measured using an anaerobic sampling
previously for arterial pressure using a calibrated (in the technique and blood gas analyzer. Blood gas analysis provides
range of measurement) and zeroed strain gauge or by mea- intermittent information; but unlike the case with PaO2
suring the rise of a column of fluid connected to a catheter measurements (discussed later), a venous sample may be
placed in the thoracic vena cava. CVP should be recorded used, allowing for easier sample acquisition. Venous values
at the end of exhalation and in the absence of positive end- are normally 3–5 mm Hg higher than arterial values.41
expiratory pressure (PEEP). CVP monitoring is not routine Hypercapnia or hypoventilation is common during
but is useful in high-risk patients. Detailed information anesthesia; in healthy mammals breathing a high fraction
regarding CVP waveforms and pulmonary artery catheter of inspired oxygen, some degree of elevation in CO2 is
monitoring is available.35 acceptable and even beneficial for cardiovascular function.
In healthy mammals, an end-tidal (exhaled breath) carbon
dioxide (ETCO2) of approximately 50–55 mm Hg or a
Monitoring the Pulmonary System PaCO2 of 60–65 mm Hg is often considered acceptable if
Ventilation the pH is maintained above 7.2 units. Interestingly, cardio-
vascular function is not well maintained in birds with the
Ventilation is the means by which the lung removes carbon elevations in CO2, so ventilation should be supported.42,43
dioxide (CO2), a product of metabolism, from the body. Although ventilators are most commonly designed for
Its regulation is important in the maintenance of acid-base animals breathing inhalation anesthetics, tools such as the
balance. For each 10 mm Hg increase in PaCO2, the pH megavertebrate demand ventilator and Hudson demand
will decrease approximately 0.05 unit. Elevated PaCO2 valve are used to support breathing and also provide oxygen.
increases cerebral blood flow, and an abnormally low PaCO2 The Ambu bag will support ventilation (with room air)
(<20 mm Hg) reduces it (in mammals). The PaCO2 has an in smaller animals. Guidelines for ventilation are based
impact on oxygenation in air-breathing animals or those on normal parameters and are provided in the section on
residing at higher elevations where the barometric pressure ventilometry and clinical assessment further on.
is low.
For most domestic species, breathing room air at sea Capnography
level (barometric pressure 760 mm Hg or torr), PaCO2 Estimates of PaCO2 may be made clinically on a continu-
is maintained between 35 and 45 mm Hg. However, ous basis using capnography (graph and value provided) or
species variations occur; domesticated cats maintain their capnometry (value only). Because the source of exhaled gas
PaCO2 at or below the low end of the range, whereas is alveolar (A) gas (which arises from blood returning to the
horses maintain values at or above the upper end of the lung from tissues where metabolic processes have occurred),
range.36,37 At higher elevations, mammalian species tend to the ETCO2 may be used to estimate alveolar and therefore
hyperventilate to varying degrees to maintain oxygen ten- arterial CO2. The relationship between alveolar (PaCO2)
sions. Limited data are available for other species.38,39 When and end-tidal ETCO2 is influenced by the breathing circuit,
interpreting blood gas (PCO2 and PO2) values, especially character and rate of breathing, and for sidestream analyzer,
from species where body temperature differs significantly the inspiratory and sampling flow rates and site of sampling.
(e.g., reptiles), the reader should consider whether values In normal clinical circumstances, the ETCO2 will be lower
should be assessed at analyzer temperature (37°C), where than the alveolar or arterial CO2 for mammals; in birds,
normal reference ranges are best defined, or at measured the relationship is less clearly defined, with reports of both
body temperature.40 Hypothermia increases solubility in higher and lower ET values.44–46 In small animals when the
the blood and therefore decreases the partial pressure; the ventilation/perfusion ratio is well maintained, the values
converse is true with a higher body temperature. Anatomic closely approximate (within 1–3 mm Hg) each other. The
CHAPTER 29 Perianesthetic Monitoring: Equipment and Interpretation 189
difference widens due to increased sampling from dead [Oxygen content (mL dL ) = (1.36 × Hemoglobin
space during certain procedures (e.g., thoracotomy).47 In concentration × % Sat 1000) + (PO2 × 0.003)]
large mammals, wide and often variable differences in the
range of 10–20 mm Hg may be observed.41 The value 1.36 refers to the oxygen-binding capacity
Two types of analyzers (sidestream and mainstream) are of 1 g of hemoglobin when well saturated. Hemoglobin is
available, each with advantages and disadvantages. Side- expressed in terms of grams per 100 mL blood; PCV/3 is a
stream analyzers are available in portable units and as part of clinical estimate. The percent saturation refers to the relative
a multiparameter physiologic monitor; with adaptation, it saturation of hemoglobin and is measured with an oximeter
may be applied to broad circumstances. Mainstream analyz- or estimated from the hemoglobin dissociation curve after
ers are suited for small animals and afford the advantage measuring the PaO2. For a PaO2 greater than 150, this is
of being battery-powered and portable. They are placed regarded as 100%. The PO2 is the partial pressure generated
between the endotracheal tube and breathing circuit; by dissolved oxygen (0.003 accounts for the solubility of
in addition to adding bulk, they may increase work of oxygen in the blood at 37°C; i.e., 0.003 mL O2 will be
breathing. dissolved in each 100 mL of blood per mm Hg PO2). From
In addition to providing information about CO2 during this formula it becomes evident that hemoglobin plays the
anesthesia maintenance, this technology may be used to major role in carrying oxygen in the blood. However, the
confirm intubation and provides valuable information driving pressure for the transport of oxygen from blood to
regarding equipment (e.g., exhausted CO2, absorbent or tissues is the partial pressure.
malfunctioning values) and circulation. Monitors are typi- The alveolar gas equation
cally equipped with alarms to alert the observer to low or PA = (PB − H2Ovap) × FiO2 − PaCO2 0.8
high CO2 readings that may result from other causes (e.g.,
apnea, hypoventilation). accounts for barometric pressure (PB), water vapor pressure
(H2Ovap), inspired oxygen (FiO2), and PaCO2. Once the
Ventilometry and Clinical Assessment alveolar pressure of oxygen has been calculated, the expected
Blood gas analyzers and capnographs are available with PaO2 may be derived. A normal alveolar (A) to arterial (a)
increasing frequency in veterinary practice, but cost issues gradient of 10 (room air) to 100 (100% oxygen) mm Hg
and lack of user comfort with the equipment still remains. may exist. Clinically a simple way to arrive at the expected
Under conditions of normal CO2 production, guidelines PaO2 is to multiply the FiO2 by 5 at sea level and by 4 at
enable the use of easily observed parameters to estimate a mile high (barometric pressure approximately 640 mm
CO2. These also guide settings of the mechanical ventilator Hg). The P/F ratio is also advocated to estimate adequacy
or demand valve. of lung function. These tools, while simple, do not take into
“Minute ventilation” (product of tidal volume and respi- account the influence of CO2, which may play a significant
ratory rate over 1 minute) approximates 150–250 mL/kg/ role.
min. For domestic mammals, tidal volume ranges from 10
to 20 mL/kg and the respiratory rate from 6 to 20 breaths Measurement of PaO2
per minute (smaller mammals maintain higher respiratory The PO2, like PCO2, is measured using a blood gas analyzer.
rates and birds usually require larger volumes due to their The sample may be obtained by percutaneous puncture
unique respiratory anatomy). Respiratory rate is obtained of an artery or sampled from an arterial catheter. Sites
by observing the rebreathing bag or the animal’s chest. Tidal for puncture or catheter placement differ by species. The
volume (two-thirds to alveolar ventilation and one-third facial, transverse facial, lateral metatarsal, dorsal pedal,
to dead space ventilation) may be estimated by excursions radial, superficial ulnar, and auricular vessels are possible
of the rebreathing bag or quantitated by a ventilometer or sites. Lingual venous sampling has been used as an alterna-
respirometer placed on the expiratory limb of the (small tive in anesthetized carnivores, as studies have shown that
animal) anesthetic breathing circuit. values closely approximate those from an arterial source.48
Variations in ventilatory management (low pressure, A consistent sample volume (commonly 1 mL, but capil-
high rate) and addition of supportive measures such as lary sampling is an option in smaller patients with some
PEEP to improve lung compliance and oxygenation may be analyzers) is collected anaerobically (to preserve accuracy
applied when deemed necessary (e.g., pneumonia). of the reading) in a heparinized syringe (to prevent blood
clots entering the machine). All air should be removed
Oxygenation from the sample and the sample corked and placed on ice
water if it is not to be run immediately; analysis should be
Oxygen delivery is the product of CO and arterial oxygen completed as soon as possible and ideally within an hour
content. Oxygen content is commonly calculated from of sampling for mammals and contemporaneously in birds.
other parameters, namely hemoglobin concentration, its An excess of heparin or contamination with air will alter
saturation (SO2), and the partial pressure of oxygen (PO2) PaO2 values (raise it for samples collected when an animal
dissolved in plasma. The relationship of these parameters to is breathing room air and lower it for animals breathing a
oxygen content is defined by the following formula: high FiO2).41,49,50
190 SE C T I O N 6 Anesthesia and Analgesia
Portable analyzers offer the veterinarian working in the generated from metabolic processes and heat dissipated
field a practical option for blood gas analysis. Many utilize (by conduction, convection, and evaporation). Anesthesia
single-use disposable self-calibrating cartridges, but these affects thermoregulatory centers in the brain and influences
may function only within certain temperature ranges or the generation and dissipation of heat. Due to a decrease
require cartridge refrigeration. in metabolic rate induced by the sleep state of anesthesia,
heat generation is decreased. Heat loss is increased by a
Measurement of SaO2 number of mechanisms related to anesthesia and surgery;
Blood gas analyzers provide intermittent information and cool intravenous fluids and inspired gases, cold tables, surgi-
may not be cost-effective. The measurement of oxygen cal clips and prepped areas, open body cavities, and so on
saturation using a pulse oximeter provides a means of con- all contribute. In general, most animals regardless of body
tinuously monitoring the patient’s oxygenation, usually at size tend to lose heat during anesthesia.
a lower cost. Additionally, the equipment is easily portable. The implications of hypothermia are many. Under
Pulse oximeter probes are of two types, transmittance and extreme conditions one may alter blood viscosity and
reflectance. The transmittance probe is attached to the coagulation and increase the likelihood of myocardial fibril-
patient externally (tongue, lips, ear, etc.), whereas the lation. Smaller decreases in body temperature will affect
reflectance probe may be placed in the oral or nasal cavity, anesthetic dose requirements (MAC is reduced 5%–8% per
rectum, vagina, inside of the eyelid, and so on. Accuracy degree centigrade decrease in body temperature) and rate of
is influenced by pigment, tissue thickness, and movement, clearance of anesthetic drugs.52
among other factors. Although hypothermia is the more likely, the opposite
Oximetry is based on the intensity of light transmitted extreme in body temperature may also be seen (as, e.g.,
through a blood sample. The different forms of hemoglobin in an animal with increased muscle activity) and is just
(oxygenated and reduced) absorb different wavelengths as consequential to the patient. Malignant hyperthermia
of light to different degrees. The pulse oximeter further is the extreme situation in which a patient may react to
distinguishes background absorption from that during the inhaled anesthetic and enter a hyperdynamic metabolic
pulsatile (arterial) flow. Accuracy is greatest in the range of state that, without early detection and intervention, is often
saturations between 80% and 95%. fatal.
Relationship of SaO2 and PaO2
Laboratory Parameters
Hemoglobin saturation is related to the PaO2 by a sigmoid
curve. Clinically, the information is analogous but the In addition to the measurement of blood gases, “blood gas”
interpretation of the numbers differs. A PaO2 of 80 mm analyzers provide additional useful measured and calculated
Hg or greater is acceptable, as is a SaO2 of 95% or greater. laboratory values. Measured values may include pH, lactate,
A PaO2 of 60 mm Hg is approximately equivalent to a glucose, creatinine, hemoglobin, and electrolytes. Bicarbon-
SaO2 of 90% for commonly anesthetized small animal ate, base excess, and saturation are derived.
species and is an indicator of hypoxemia; variations in this The range (7.35 and 7.45 units) for normal pH (at
relationship likely exist in nondomestic species but are not 37°C, approximating normal human body temperature)
fully elucidated.41,51 Due to the nature of this relationship, is well described for mammals. A pH below this range
an SaO2 of 100% (hemoglobin maximally saturated) could indicates acidemia and one above this range, alkalemia.
reflect a PaO2 ranging from 100 mm Hg to upward of Carbon dioxide (as previously described) and bicarbonate
500 mm Hg. Clinically, this becomes limiting if the goal (20–28 mEq/L) describe the respiratory and metabolic
is to assess pulmonary function in the face of high FiO2. contributions. As with carbon dioxide, bicarbonate
values vary between mammalian species; carnivores tend
Monitoring Body Temperature to present with lower (17–24 for cats) values, whereas
herbivores often have higher (24–32 for horses) values.
Although simple to perform either by intermittent monitor- Bicarbonate provides an indication of the metabolic con-
ing (using a thermometer placed in either the rectum or tribution to pH (high values indicate metabolic alkalosis
auricular canal) or by continually using a thermistor probe and low values metabolic acidosis). Bicarbonate values are,
(placed in the esophagus or rectum), this is often ignored however, influenced by carbon dioxide (as described by
in clinical practice. Knowledge of normal ranges for the the Henderson-Hasselbalch equation). Broadly, an increase
different species is critical for appropriate interpretation. in PaCO2 of 10 mm Hg will result in an increase of
For example, normal body temperature in many avian 1–3 mEq/L in the bicarbonate value; note that this is not
species would be considered hyperthermic for mammals. compensation but simply the result of a mass shift. Base
Conversely some marsupials (e.g., kangaroo, wallaby) and excess provides a true measure of the acid-base balance by
most reptiles have normal body temperatures that are lower removing the influence of CO2 on bicarbonate. Base excess
than those of mammals. values (mEq/L) tend to be more negative in carnivores
Body temperature is controlled by thermoregula- (−7 to +3) and more positive (0 to +4) in herbivores.
tory centers in the brain and reflects the balance of heat A negative value outside these ranges indicates metabolic
CHAPTER 29 Perianesthetic Monitoring: Equipment and Interpretation 191
acidosis, whereas a positive value indicates metabolic administration in dogs, J Am Vet Med Assoc 239(1):81–89,
alkalosis. 2011.
Lactate and blood glucose values should be interpreted 10. Ducharme NG, Fubini SL: Gastrointestinal complications
associated with the use of atropine in horses, J Am Vet Med Assoc
based on typical normal ranges. A lactate value greater than
182(3):229–231, 1983.
2 mmol/L indicates anaerobic metabolism and a blood 11. Lerche P: Anticholinergics. In Grimm KA, Lamont LA, Tran-
glucose value less than 60 mg/dL points to hypoglycemia. quilli WJ, et al, editors: Veterinary anesthesia and analgesia, ed 5,
Electrolyte values may vary slightly among species but tend Ames, IA, 2015, John Wiley & Sons Inc, pp 178–182.
to be held to fairly tight ranges; ionized values for potassium 12. Olson ME, Vizzutti D, Morck DW, et al: The parasympatholytic
and calcium are influenced by pH (a low pH will falsely effects of atropine sulfate and glycopyrrolate in rats and rabbits,
elevate both values) and should be interpreted in light Can J Vet Res 57:254–258, 1993.
of this. 13. Lin H: Comparative anesthesia and analgesia of ruminants and
In animals that are anemic or at risk for hemorrhage, swine. In Grimm KA, Lamont LA, Tranquilli WJ, et al, editors:
measurement of the packed red cell volume and total Veterinary anesthesia and analgesia, ed 5, Ames, IA, 2015, John
protein provides additional useful information. Wiley & Sons Inc, pp 743–753.
14. Caulkett NA, Cattet MR, Cantwell S, et al: Anesthesia of wood
bison with medetomidine-zolazepam/tiletamine and xylazine-
Summary zolazepam/tiletamine combinations, Can Vet J 41:49–53, 2000.
15. Giral M, García-Olmo DC, Gómez-Juárez M, et al: Anaesthetic
Although not an exhaustive list, this chapter provides effects in the ferret of alfaxalone alone and in combination
options for monitoring animals during heavy sedation and with medetomidine or tramadol: a pilot study, Lab Anim
general anesthesia. Based on working conditions and degree 48(4):313–320, 2014.
of compromise, the anesthetist may select from these to 16. Kiers HD, Hofstra JM, Wetzels JF: Oscillometric blood pres-
complement subjective observations. Appropriate use and sure measurements: differences between measured and calculated
interpretation are key to correctly applying the information mean arterial pressure, Neth J Med 66(11):474–479, 2008.
to benefit the animal. 17. Babbs CF: Oscillometric measurement of systolic and diastolic
blood pressures validated in a physiologic mathematical model,
Biomed Eng Online 11:56, 2012.
References 18. Scansen BA, Vitt J, Chew DJ, et al: Comparison of forelimb
and hindlimb systolic blood pressures and proteinuria in healthy
1. Murrell JC, Hellebrekers LJ: Medetomidine and dexmedeto- Shetland Sheepdogs, J Vet Intern Med 28(2):277–283, 2014.
midine: a review of cardiovascular effects and antinociceptive 19. Haberman CE, Kang CW, Morgan JD, et al: Evaluation of oscil-
properties in the dog, Vet Anaesth Analg 32:117–127, 2005. lometric and Doppler ultrasonic methods of indirect blood pres-
2. Grimsrud KN, Ait-Oudhia S, Durbin-Johnson BP, et al: sure estimation in conscious dogs, Can J Vet Res 70(3):211–217,
Pharmacokinetic and pharmacodynamic analysis comparing 2006.
diverse effects of detomidine, medetomidine, and dexmedeto- 20. Stepien RL, Rapoport GS: Clinical comparison of three methods
midine in the horse: a population analysis, J Vet Pharmacol Ther to measure blood pressure in nonsedated dogs, J Am Vet Med
38(1):24–34, 2015. Assoc 215(11):1623–1628, 1999.
3. Sinclair MD: A review of the physiological effects of α2-agonists 21. Petric AD, Petra Z, Jerneja S, et al: Comparison of high definition
related to the clinical use of medetomidine in small animal oscillometric and Doppler ultrasonic devices for measuring blood
practice, Can Vet J 44(11):885–897, 2003. pressure in anaesthetised cats, J Feline Med Surg 12(10):731–737,
4. KuKanich B, Wiese AJ: Opioids. In Grimm KA, Lamont LA, 2010.
Tranquilli WJ, et al, editors: Veterinary anesthesia and analgesia, 22. Aarnes TK, Hubbell JA, Lerche P, et al: Comparison of invasive
ed 5, Ames, IA, 2015, John Wiley & Sons Inc, pp 207–226. and oscillometric blood pressure measurement techniques in
5. Jacobson ER, Kollias GV, Heard DJ, et al: Immobilization anesthetized camelids, Can Vet J 53(8):881–885, 2012.
of African elephants with carfentanil and antagonism with 23. Harvey L, Knowles T, Murison PJ: Comparison of direct and
nalmefene and diprenorphine, J Zoo Wildl Med 19(1–2):1–7, Doppler arterial blood pressure measurements in rabbits during
1988. isoflurane anaesthesia, Vet Anaesth Analg 39:174–184, 2012.
6. Buss P, Miller M, Fuller A, et al: Cardiovascular effects of etor- 24. Sant Cassia EV, Boswood A, Tordiffe AS: Comparison of
phine, azaperone, and butorphanol combinations in chemically high-definition oscillometric and direct arterial blood pressure
immobilized captive white rhinoceros (Ceratotherium simum), J measurement in anesthetized cheetahs (Acinonyx jubatus), J Zoo
Zoo Wildl Med 47(3):834–843, 2016. Wildl Med 46(3):506–516, 2015.
7. Mans C, Guzman DS-M, Lahner LL, et al: Sedation and physi- 25. Tünsmeyer J, Hopster K, Feige K, et al: Agreement of high
ologic response to manual restraint after intranasal administra- definition oscillometry with direct arterial blood pressure
tion of midazolam in Hispaniolan Amazon parrots (Amazona management at different blood pressure ranges in horses under
ventralis), J Avian Med Surg 26(3):130–139, 2012. general anaesthesia, Vet Anaesth Analg 42(3):286–291, 2015.
8. Citino SB, Bush M: Reference cardiopulmonary physiologic 26. Gladczak AK, Shires PK, Stevens KA, et al: Comparison of
parameters for standing, unrestrained white rhinoceroses (Cera- indirect and direct blood pressure monitoring in normotensive
totherium simum), J Zoo Wildl Med 38(3):375–379, 2007. swine, Res Vet Sci 95(2):699–702, 2013.
9. Congdon JM, Marquez M, Niyom S, et al: Evaluation of the 27. Van Citters RL, Kemper WS, Franklin DL: Blood flow and
sedative and cardiovascular effects of intramuscular administra- pressure in the giraffe carotid artery, Comp Biochem Physiol
tion of dexmedetomidine with and without concurrent atropine 24:1035–1042, 1968.
192 SE C T I O N 6 Anesthesia and Analgesia
28. Chinnadurai SK, Wrenn A, DeVoe RS: Evaluation of noninva- and analgesia, ed 5, Ames, IA, 2015, John Wiley & Sons Inc,
sive oscillometric blood pressure monitoring in anesthetized boid pp 86–113.
snakes, J Am Vet Med Assoc 234(5):625–630, 2009. 42. Seok S, Jeong D, Hong I, et al: Cardiorespiratory dose-response
29. Johnston MS, Davidowski LA, Rao S, et al: Precision of repeated, relationship of isoflurane in Cinereous vulture (Aegypius monachus)
Doppler-derived indirect blood pressure measurements in con- during spontaneous ventilation, J Vet Med Sci 79(1):160–165,
scious psittacine birds, J Avian Med Surg 25(2):83–90, 2011. 2017.
30. Schier MF, Raisis AL, Secombe CJ, et al: Effects of dobutamine 43. Ludders JW, Rode J, Mitchell GS: Isoflurane anesthesia in sand-
hydrochloride on cardiovascular function in horses anesthetized hill cranes (Grus canadensis): minimal anesthetic concentration
with isoflurane with or without acepromazine maleate premedi- and cardiopulmonary dose-response during spontaneous and
cation, Am J Vet Res 77(12):1318–1324, 2016. controlled breathing, Anesth Analg 68(4):511–516, 1989.
31. Rosati M, Dyson DH, Sinclair MD, et al: Response of hypo- 44. Desmarchelier M, Rondenay Y, Fitzgerald G, et al: Monitor-
tensive dogs to dopamine hydrochloride during deep isoflurane ing of the ventilatory status of anesthetized birds of prey by
anesthesia, Am J Vet Res 68(5):483–494, 2007. using end-tidal carbon dioxide measured with a microstream
32. Pascoe PJ, Ilkiw JE, Pypendop BH: Effects of increasing infusion capnometer, J Zoo Wildl Med 38(1):1–6, 2007.
rates of dopamine, dobutamine, epinephrine, and phenylephrine 45. Pettifer GR, Cornick-Seahorn J, Smith JA, et al: The comparative
in healthy anesthetized cats, Am J Vet Res 67(9):1491–1499, cardiopulmonary effects of spontaneous and controlled ventila-
2006. tion by using the Hallowell EMC Anesthesia WorkStation in
33. Gosliga JM, Barter LS: Cardiovascular effects of dopamine Hispaniolan Amazon parrots (Amazona ventralis), J Avian Med
hydrochloride and phenylephrine hydrochloride in healthy Surg 16(4):268–276, 2002.
isoflurane-anesthetized New Zealand White rabbits (Oryctolagus 46. Edling TM, Degernes LA, Flammer K, et al: Capnographic
cuniculus), Am J Vet Res 76(2):116–121, 2015. monitoring of anesthetized African grey parrots receiving
34. Pascoe PJ, Ilkiw JE, Stiles J, et al: Arterial hypertension associated intermittent positive pressure ventilation, J Am Vet Med Assoc
with topical ocular use of phenylephrine in dogs, J Am Vet Med 219(12):1714–1718, 2001.
Assoc 205(11):1562–1564, 1994. 47. Wagner AE, Gaynor JS, Dunlop CI, et al: Monitoring adequacy
35. Reems MM, Aumann M: Central venous pressure: principles, of ventilation by capnometry during thoracotomy in dogs, J Am
measurement, and interpretation, Compend Contin Educ Vet Vet Med Assoc 212(3):377–379, 1998.
34(1):E1, 2012. 48. Wagner AE, Muir WW, Bednarski RM: A comparison of arterial
36. Grandy JL, Steffey EP, Miller M: Arterial blood PO2 and PCO2 and lingual venous blood gases in anesthetized dogs, J Vet Emerg
in horses during early halothane-oxygen anaesthesia, Equine Vet Crit Care 1(1):14–18, 1991.
J 19(4):314–318, 1987. 49. Rezende ML, Haskins SC, Hopper K: The effects of ice-water
37. Waddell LS: Blood gas analysis. NAVC Clinician’s Brief January storage on blood gas and acid-base measurements, J Vet Emerg
2012:18–19. Crit Care 17(1):67–71, 2007.
38. Stinner JN: Ventilation, gas exchange and blood gases in the 50. Hopper K, Rezende ML, Haskins SC: Assessment of the effect
snake, Pituophis melanoleucus, Respir Physiol 47(3):279–298, of dilution of blood samples with sodium heparin on blood
1982. gas, electrolyte, and lactate measurements in dogs, Am J Vet Res
39. Schnellbacher R, da Cunha A, Olson EE, et al: Arterial catheter- 66(4):656–660, 2005.
ization, interpretation, and treatment of arterial blood pressures 51. Haymerle A, Knauer F, Walzer C: Two methods to adapt the
and blood gases in birds, J Exot Pet Med 23(2):129–141, 2014. human haemoglobin-oxygen dissociation algorithm to the blood
40. Wang T, Abe AS, Glass ML: Temperature effects on lung and of white rhinoceros (Ceratotherium simum) and to determine the
blood gases in Bufo paracnemis: consequences of bimodal gas accuracy of pulse oximetry, Vet Anaesth Analg 43(5):566–570,
exchange, Respir Physiol 113(3):231–238, 1998. 2016.
41. Haskins SC: Monitoring anesthetized patients. In Grimm KA, 52. Sessler DI: Temperature monitoring and perioperative thermo-
Lamont LA, Tranquilli WJ, et al, editors: Veterinary anesthesia regulation, Anesthesiology 109(2):318–338, 2008.
SECTION 7
Diagnostics
30 Wildlife Necropsy Primer, 194
193
30
Wildlife Necropsy Primer
DENISE MCALOOSE
194
CHAPTER 30 Wildlife Necropsy Primer 195
• BOX 30.1 Necropsy Examination Personal Protective Equipment (PPE), Supplies, Equipment, and Tools
Documentation • Culture: sterile in-date swabs, urine cups, and bags, culture
• Forms: necropsy protocol and form, morphometrics data transport media/tubes (for bacteria, virus)
sheets, tissue sample checklist, human interaction form, • Tissue fixation: 10% neutral buffered formalin, 4%
notebook/paper (regular and waterproof [e.g., Rite in the glutaraldehyde or other EM fixative in small screw-top vials,
Rain®]) isopropyl alcohol (for ectoparasites and endoparasites,
• Labeling: tape, laundry tags with metal clips, pencils, cytologic preparations, etc.)
waterproof marking pens and pencils (to label samples that • For genetic/molecular diagnostics (aliquot into small screw-
will be immersed in liquid fixatives), Tyvek®* top vials): 20% DMSO/saturated saline solution (genetics),
• Photodocumentation: digital or film camera, batteries, RNA-later® or TRIzol® (molecular diagnostics)
memory cards, labels (include in every image) • Lighting: head lamp, flashlight, batteries, light bulbs,
generator with extra bulbs and fuel
Safety and Basic PPE • Cold chain: ice chest, ice packs, refrigerator, freezer (−20°C,
−80°C, dry ice, liquid nitrogen), absorbent packing materials
• Clothing: gloves, mask (e.g., surgical, N95, masks with
• Laboratory equipment: microscope (for field settings: field
integrated face shield), eye protection (goggles, face shield),
adapted [mirror as a light source], car battery–adapted power
surgical scrubs, lab coat, coveralls, aprons, boots, gloves,
source, generator), centrifuge
caps (head cover)
• Other: ropes/straps/chains, string/suture, parafilm, glass
• Disinfection: sponges, dish soap, scrub brushes, disinfectant,
slides and slide boxes, water supply/source (for cleaning/
bleach, alcohol (70% ethyl alcohol)
clean-up), plastic tarps, plastic tape/ropes to cordon off
• Other: first aid kit, communication link (e.g., satellite phone),
necropsy site, garbage bags, biohazard bags, disinfectant,
SDS/MSDS
bleach, sponges/scrub brushes, paper towels, portable
Tools generator (for electric powered saws, refrigerators, etc.)
• Cutting: scalpel blades, scalpel handles, knives (6- or 8-in Additional Equipment for Small Carcasses (<5 g)
blade), knife sharpeners†, scissors (small and large), bone
• Magnification: dissecting microscope, magnification
shears, handsaws (e.g., hacksaw, reciprocating saw),
headband, surgical loupe
axe/hatchet, wedges, mallet/hammer, cutting boards,
• Cutting: microdissection forceps, scissors
rongeurs, loppers (hedge clippers), chisel/wedge (e.g.,
T shaped) Additional Equipment for Megavertebrate (whale,
• Tissue handling: forceps, meat hook elephant, etc.)
• Containers: rigid leakproof wide-mouth spillproof screw-top
containers (various sizes), zip-top plastic bags and Whirl- • Cutting: flensing knives, large reciprocating saw, large
pak® (various sizes), serum tubes for fluid, blood, and urine hammer/mallet and chisel, appropriate sharpener(s), shovel
collection, aluminum foil, Teflon® bags, cryovials, sterile vials/ • Tissue handling: large meat hooks, gaff hook
containers/bags, sterile needles and syringes (various sizes), • Morphometrics (metric): 20 m long (min) tape measure
trochar (various sizes) • Mobility (for moving carcass, appendages, etc.): hoist/crane
• Morphometrics (metric): ruler, calipers, tape measure, scales with large-capacity mounted scale
• Sterilization: sterile instruments, matches or propane torch • Containers: large sealable containers (e.g., vials to garbage
and searing blade/spatula, isopropyl alcohol for flaming cans)
instruments • Other: thick rope/chain (min of 20 m long), block and tackle
*When labeled with permanent marker or pencil, may be placed in liquid (e.g., formalin) for identification purposes.
†
For standard knives, recommend hand held sharpeners not sharpening stones because the use of stones requires significant expertise, and inexperienced operators
may dull rather than sharpen cutting instruments (e.g., knives, scissors).
This is a general guide that may be tailored to meet the specific situational needs of a particular necropsy examination(s).
MSDS, Material safety data sheets; PPE, personal protective equipment; SDS, safety data sheets.
Data from http://www.seadocsociety.org/wp-content/uploads/Orca-necropsy-protocol-FINAL-May-15-2014.pdf; Necropsy procedures for wild animals. In White L,
Edwards A, editors: Conservation research in the African rain forests: a technical handbook, New York, 2000, Wildlife Conservation Society, pp 196–217; Woodford
M, Keet D, Bengis R: A guide to post-mortem procedures and a review of pathological processes identified in the elephant. In Woodford M, editor. Post-mortem
procedures for wildlife veterinarians and field biologists. Paris, France, 2000, Office International des Epizooties, Care for the Wild and the Veterinary Specialist
Group/Species Survival Commission of the World Conservation Union (IUCN).
identify problems that pose safety risks such as cold or regionally, or internationally reportable/notifiable disease
rain, rising tides, or suboptimal animal position; in some (e.g., USDA [US Department of Agriculture], CDC
cases where safety cannot be ensured, a necropsy should be [Centers for Disease Control and Prevention], OIE [World
postponed or abandoned. Wildlife and veterinary patholo- Organization for Animal Health] reportable disease) or
gists and clinicians and human medical health professionals illegal activities (e.g., poaching, poisoning) are suspected,
are typically responsible for making these decisions based on the scene should be secured and appropriate authorities
a risk assessment that relies on an understanding of specific contacted before proceeding. For reportable/notifiable
and relative risk, knowledge about infectious diseases and diseases, direct contact with regulatory agencies or online
circumstantial situations, institutional standard operating information will provide the most up-to-date information
protocols, and best practice protocols. In addition, if at about those diseases that are listed (see http://www.oie.int/
any point there is a suspicion or confirmation of a locally, animal-health-in-the-world/oie-listed-diseases-2017/).
196 SE C T I O N 7 Diagnostics
In advance of a necropsy, it is important to have organized of a struggle) that may be relevant to the circumstances of
plans and assign roles and responsibilities. Standard tasks the animal’s death.
include identification of a lead prosector/pathologist, and
individuals or teams to collect morphometric data, perform External and Internal Examination
prosection, label and manage samples (diagnostic, research,
archival), perform photodocumentation, manage data/data A good rule of thumb before performing a necropsy pro-
sheets (record, upload, organize), and ensure fulfillment cedure is to familiarize yourself with the normal anatomy
of all protocols (including sample collection for research and natural history of the species on which you will be
requests), ship samples, perform clean-up/disinfection and working (e.g., normal diet, fecal consistency, color patterns,
carcass disposal, and manage communication within and sexual dimorphism, common and zoonotic diseases). This
between necropsy investigative team members and stake- is particularly important when performing a necropsy on a
holders, including the media. Having organized plans and species with which you are not familiar (e.g., what is the best
assigned roles is particularly important in megavertebrate approach to opening the body cavity of a chambered nau-
necropsies or mass mortality events, due to the size or tilus [Nautilus pompilius]) and will inform activities related
number of animals, participants of varied skill levels, and to PPE and risk. Numerous texts and online resources, as
numerous different agencies that may be involved. well as colleagues with species-specific knowledge, are good
places to start. It is also worth remembering that, although
Necropsy Examination every taxon and species has unique features, familiarity with
one often serves as a helpful reference for another. For
Before a postmortem examination begins, it is worth example, familiarity with the anatomy of well-described
remembering that you, whatever your role, are the most and studied domestic or farmed species is directly applicable
valuable asset in the process. All conclusions drawn in a to the anatomy of most nondomestic mammals, fish, and
mortality investigation rest on the information and samples birds. Invertebrates provide a greater challenge. This is an
you and your team collect. Best intentions and memory are incredibly diverse group that includes more than 99% of all
no substitute for the collection of real-time verbal, written, animal species on the planet. Over the past decade, there
and photographic documentation. Each person involved in has been increased interest in this group, and expertise and
the necropsy must take personal responsibility for his or resources are becoming more widely available.
her actions. Outcomes are as good as the data you collect All necropsy examinations should be systematic, follow
and the focus you bring to the task, and what you do standardized procedures and protocols, and include check-
during and after the examination will make the impossible, lists for the collection of “standard” sets of tissues (Box
possible. 30.2). This ensures that a carcass is thoroughly examined,
All mortality investigations, regardless of whether they standardized information for generating baseline and
involve a single animal or a large mortality event, contain a documenting abnormalities is captured, and a complete
set of consistent elements: a systematic, iterative diagnostic set (or sets) of data and samples are collected for baseline,
plan that applies evidence against a set of case definitions diagnostic, archival, and research purposes. When practical,
and differential diagnoses to establish a cause of death (Fig. fulfilling sampling/research-associated requests (e.g., bio-
30.1). Central in this process is the necropsy examina- materials requests) allows valuable, additional information
tion. Providing specific necropsy protocols for all taxa and to be generated. Where applicable, standardized necropsy
for every situation is beyond the scope of this chapter. procedures and/or sampling/research-associated requests
However, all necropsy procedures follow a similar, basic that are required by governmental agencies should also be
format (described below). followed.
Even in this age of digital photography, written descrip-
Collection and Documentation of Relevant tions of visual observations (animal, physical environment,
Historical Information etc.) remains a critically important component of a necropsy
examination. But that does not mean that they need to be
In addition to information that will be collected from the overly complicated. Two general rules of thumb during a
carcass, it is quite important to collect and record relevant necropsy: describe what you see and limit descriptions to
death-related information. This includes basic animal infor- factual observations rather than interpretations or diagnoses
mation (e.g., species, breed, gender, age/age group, birth (this occurs later). Keep in mind that you describe all sorts of
and death dates) and clinical medical information about things every day and the language you use to describe what
the subject animal, contact animals, and other sympatric you see during a necropsy examination is no different than
species (including other wildlife/vermin, humans, domestic the language you use during everyday conversations. Basic
animals, livestock, etc.), environmental, and epidemiologic necropsy descriptions include information about location
information. It is also important to record information (e.g., organ, body system) number (e.g., 1, 10, more than
about the location in which an animal was found dead, 20 but less than 100, multiple, many, myriad), color, size,
environmental conditions/enclosure information, hus- shape, distribution (e.g., focal, regionally extensive, diffuse),
bandry changes, and to note other findings (e.g., evidence consistency and texture (soft, firm, hard), and odor. For
CHAPTER 30 Wildlife Necropsy Primer 197
• Figure 30.1 Mortality Investigation Flow Chart. All mortality investigations include a consistent set of
standardized activities. These include collection of information related to the circumstances of an animal’s
death, gross necropsy and sample collection for ancillary diagnostic testing, and establishing a set of
differential diagnoses and case definitions against which test results are applied in order to arrive at final
diagnoses and conclusions. Integral in this process is collection of information and documentation of all
findings and clear and consistent communication with all relevant stakeholders.
example, “Dozens of pinpoint to 5 mm in diameter, soft, collected samples should be entered into logs and secure
light tan nodules that contain thick, white material on cut databases developed for this purpose. This is important
section are present throughout the liver.” Measurements are for a whole host of reasons, including rapid data retrieval
useful for documenting lesions, including weights of organs and collation during mortality events, and data and sample
thought to be too large or too small. Size measurements sharing for current and future research projects. Information
should be taken in three dimensions (length × width × should be as detailed as possible to ensure that important
height/thickness). Remember, a description should include information is not forgotten or lost.
those details that allow someone who did not witness the Under ideal conditions, a necropsy is performed and
necropsy to create an accurate image of what you saw. Also samples are collected from freshly dead animals. If necropsy
remember that a description is not an interpretation or a will be delayed, carcasses should be refrigerated or kept cool
diagnosis. Those are best left to the pathologist to formulate on ice for several hours to a few days; freezing should be an
based on your description (in the above description, it is option of last resort because the freeze-thaw cycle will alter
hoped that you imagined multiple hepatic abscesses). For the color of many tissues and kill many pathogens (e.g.,
photodocumentation, always include a size reference (e.g., bacteria, fungi), and ice crystal formation will significantly
small ruler) and animal-related information (minimum: limit the value of histology. However, under some circum-
ID number, date). All information from the necropsy and stances (e.g., when a necropsy cannot be safely, thoroughly,
198 SE C T I O N 7 Diagnostics
or adequately performed in a timely manner), a necropsy findings in the structures of the head (e.g., eyes, ears, nares,
can be performed on previously frozen carcasses, although bill, beak, teeth, cere, gills, tonsil); the outer covering of
the results will not be optimal. the body (e.g., skin, hair, feathers, scales, claws, hooves);
The external examination is an assessment of the tissues the subcutis, glands (e.g., periorbital, salivary, scent, salt,
outside the main body cavities. This begins with direct perianal, uropygial), and skeletal muscles; external repro-
measurement or estimation of carcass weight, collection of ductive system tissues; the skeletal system (including one to
morphometric data, photodocumentation, and assessment multiple joints); and the peripheral nervous system. It also
of state of decomposition. Every carcass, regardless of its includes objective (measured) and/or subjective assessment
state of decomposition, may provide valuable scientific data. of fat stores (e.g., subcuticular, blubber) and muscle mass
In many field situations, access to fresh carcasses is the and collection of bone marrow (typically femur or rib in
exception rather than the rule. Environmental conditions nonavian and tibiotarsal bone in avian species). Written
(e.g., high humidity, high temperatures, or dense foliage and photographic documentation of all notable normal and
in natural or recreated settings) and interval from death abnormal findings should be recorded, and representative
to discovery all affect the degree of decomposition. As the sample(s) from all tissues and all lesions should be collected
body decomposes, all tissues degrade. The speed at which and placed into 10% neutral buffered formalin (NBF) for
this occurs and the reliability of diagnostic test results that histology or collected as appropriate for ancillary diagnostic
may be obtained from different tissue types during this testing.
process vary. It is therefore important to document the An internal examination is an assessment of the organ
degree of carcass decomposition, otherwise known as carcass systems and individual organs in the body cavities. There
condition, to contextualize observations and diagnostic are a number of common approaches to an internal exami-
test results. This is a subjective assessment, and a relatively nation. One is not necessarily better than another, and
common scheme categorizes postmortem carcass condition your preferred method may vary depending on the species
as excellent (freshly dead) to mildly autolyzed, moderately and specific circumstances. Regardless of the method, of
autolyzed, severely autolyzed, or skeletonized/desiccated/ paramount importance is that you work safely, have an
mummified remains. In the marine mammal community, organized plan, and have a systematic strategy for organ
several protocols use a similar carcass condition coding and organ system examination to ensure all tissues are
system.3 Assessment of carcass condition is particularly examined, lesions documented, and samples collected. This
useful in low-resource or time-restricted settings to identify is especially important when working in large teams, on
those carcasses that will be the most diagnostically valuable large animals, or in complicated situations (e.g., a mass
(e.g., those that are “freshest” or most intact) and to direct mortality event). It is also important to work cleanly to
sampling and testing. minimize, as much as possible, contamination of the work-
In vertebrates a systematic external examination of site and exposure risks to personnel and other animals (e.g.,
the carcass includes evaluation of normal and abnormal scavengers).
CHAPTER 30 Wildlife Necropsy Primer 199
expanded scope and scale, that is inherent in an outbreak/ necropsies. Establishment of a sampling and necropsy
mortality event. In many countries, local and national plans protocol as soon as possible at the outset of the outbreak
or other strategies have been developed and are activated or mortality event is crucial to obtaining the best data to aid
in outbreak situations. Familiarity with those that exist in the investigation. Sampling and necropsy protocols should
your region is important because valuable resources that aid incorporate appropriate tissue collection for evaluation
or are necessary in your response and investigation may be of not only the first tier differential etiologies but also a
available through these programs. For example, the Incident broad range of alternative etiologies. Often with large-scale
Command System (ICS) is a management concept that was mortality events, it is not practical or logistically possible to
created to address the demands and complexities inherent examine or sample every carcass in a reasonable postmortem
in emergency response or natural disaster events. In the interval. As such, focusing thorough diagnostic efforts on
United States the ICS is used by many federal agencies, the freshest carcasses, especially those that die earliest in
and its adapted use may provide organizational structure to the event, may yield the most reliable data. In addition,
veterinary outbreak and mass mortality event response and plans should be established at the outset of the investigation
investigations. Main components of the ICS are: Unified for carcass management, database and sample management
Command, Operations, Planning, Logistics, and Finance and storage, and sharing of data. It is also valuable during
and Administration; subdivision units (e.g., incident com- the course of the event to reevaluate the plan in order to
mander; safety, information and liaison officers) can be make modifications based on what is being learned during
developed depending on the needs for the particular event the event (e.g., recognition of broader geographic scale
(see https://training.fema.gov/EMIWeb/IS/ICSResource/ or additional affected species) that increase efficiency and
index.htm; accessed 16 Feb 2018). investigation outcomes.
Critical components in mortality events and outbreak
investigations include (1) identifying an investigative team Clean-Up and Carcass Disposal
and resources, (2) verifying that an outbreak is occurring, As occurs during a necropsy, PPE should be worn during
(3) establishing a case definition(s) and categorizing cases clean-up and carcass disposal. Blood and residual tissues
using results of necropsy examination, histology, ancillary should be removed from instruments and work surfaces
diagnostic test results, etc., to establish the scope of the with soap and water, after which they should be disinfected.
outbreak, (4) establishing baseline information about the The best choices for disinfection have broad antimicrobial
event and disease (e.g., temporospectral, species, gender, properties (e.g., 0.5% sodium hypochlorite; 10% bleach,
age class), (5) examining the descriptive epidemiologic 70% alcohol, borax; see also references [14 and 15] for
features of the cases to generate hypotheses, (6) testing more information about common disinfectants) and are
hypotheses and performing additional analysis as needed, effective against many common pathogens. Depending on
(7) implementing control measures (when possible), and (8) the disinfectant (e.g., bleach), instruments may need to be
communicating findings and maintaining surveillance.12,13 rinsed after disinfection to prevent corrosion. It should be
For any event, it is also important to quickly determine if noted that prions are resistant to a number of common
a disease (e.g., infectious, toxic) with significant zoonotic disinfectants and other forms of destruction.16 Current, best
or animal health impacts (e.g., anthrax, Ebola) or illegal practice protocols for disinfection and carcass management
activities is present in order to engage and work together should be obtained and implemented in consultation with
with the appropriate agencies and authorities in the mortal- governmental agencies for suspected or confirmed cases.
ity investigation. It is also important that the investigative Whether in a laboratory or a field setting, necropsy
team has a broad range of expertise (e.g., epidemiology, examinations and disposal of carcass, necropsy waste,
population biology, pathology, clinical medicine, genetics, sharps, and infectious materials should be performed in a
bacteriology, virology, toxicology, and species- and environ- manner that minimizes environmental contamination and
mental specific expertise) to help guide the investigation. exposure of the necropsy team or domestic or wild animals
Pathologists often work closely with epidemiologists during to infectious, toxic, or other disease agents. Efforts should
outbreak or mass mortality event investigations to construct be made to minimize the amount of nonanimal waste (e.g.,
case definitions, develop hypotheses on the cause of the aprons, masks, gloves, plastic bags) that is generated and
event, analyze data, recommend ancillary diagnostic testing, burned, buried, or transported. It is important to be aware
and ultimately when making recommendations for control of and adhere to relevant organizational, local, national,
measures or long-term surveillance. and international regulations and protocols for carcass and
Epidemiologic characteristics of an outbreak, along with medical waste disposal, which may involve consultation
historic data and necropsy findings, guide decisions related with local environmental and health agencies. Options may
to first tier diagnostic testing (those deemed most relevant be limited due to a number of factors, including personal
and immediately important). First tier diagnostic tests and safety, location, size of a carcass(es), environmental condi-
histopathology findings often guide further ancillary testing. tions, location relative to water sources/catchments/wells
Approaches to necropsy evaluation and sampling during etc., ability to safely move/relocate a carcass for disposal
an outbreak or large-scale mortality event are like those generally and within a reasonable timeframe (this may
used during routine, systematic, and thorough diagnostic be logistically and politically complex and challenging),
CHAPTER 30 Wildlife Necropsy Primer 201
financial limitations, animal welfare and environmental Collecting a limited set of tissues may provide a diagnosis.
considerations, and willingness/ability/permission of con- However, this may reflect an incomplete picture of the
tracting facilities/landfills, etc. to accept the carcass/waste. cause and contributing or predisposing factor(s) in death.
Options for carcass, tissue, and waste disposal include In a worse-case scenario, this can lead to misinterpretations
incineration/burning and burial, burial, rendering, landfill, and false conclusions that drive inappropriate husbandry,
fermentation, and biocremation (alkaline hydrolysis), which medical management, or other animal- or outbreak-related
may be possible on-site or by external contractors, as well activities and mitigation strategies.
as mounding, composting, and natural decay at the site of Sample handling and storage for histology and ancil-
death/necropsy. A number of detailed descriptions of several lary diagnostic testing are critically important to optimize
of these options are available and include those of national their diagnostic value and contributions to animal health,
and international governmental and nongovernmental management, surveillance, and conservation outcomes (see
regulatory agencies.4,5,17 On-site incineration/burning and Table 30.1). With the exception of formalin-fixed tissues,
burial or burial alone may be the most practical option in most diagnostic samples need to be refrigerated or frozen to
some cases (e.g., in mass mortality events). In known or prevent degradation and maintain the viability of pathogens,
suspected anthrax cases, incineration/burning and burial toxins, etc. if immediate on-site testing is not available,
or burial, or other methods that prevent sporulation and for transport to an external diagnostic laboratory, or when
destroy the bacteria, should be used.2 If prion disease is archived. This often includes establishing and maintaining
suspected, a 2001 report suggested rendering, incineration, a cold chain, which can be particularly challenging in field
and alkaline hydrolysis (all under certain conditions) as settings.18 This involves appropriate, consistent temperatures
the most reliable technologies at that time for reducing using one or a combination of materials and equipment.
the infectivity of the organism.4 In addition, remember Storage containers/equipment holding diagnostic samples
that pathogens may persist, proliferate, and pose extended should not be used for holding food, beverages, or other
human and animal exposure risks in unburned, buried or medical materials such as pharmaceuticals. Basic equipment
closed plastic bags. typically includes refrigerators (4°C ice chests/boxes, ice/gel
packs, etc.), freezers (e.g., 0°C, −20°C, −80°C; non “frost
Sample Collection and Management free”), and containers appropriate for standard and ultralow
storage (e.g., cryovials). Other options include dry ice and
Even when the cause of death seems obvious, it is important liquid nitrogen (LN2) (e.g., dewars [vacuum flasks], dry
that interpretations and conclusions drawn during a mortal- shippers [vapor shippers]). Use of and handling samples
ity investigation on an individual or group of animals are stored at ultralow temperatures should be carried out with
informed by all relevant information. This process is sequen- extreme caution because exposure can cause severe burns.
tial and iterative, with gross necropsy typically followed In additional, dry ice and LN2 vapor can cause asphyxiation
by histology. Both directly affect decisions about ancillary and can be explosive so should be used only in ventilated
diagnostic tests that are needed to confidently arrive at spaces or containers; handling should be limited to trained
conclusions and are subsequently performed. In addition to personnel. For electrical equipment, a consistent source of
a thorough, systematic necropsy examination, collection of power is an absolute necessity. In all settings, backup sources
a set of tissues for histology and ancillary diagnostic testing of electricity, often in the form of a generator, are essential.
and tissue archiving (Table 30.1) should be a standard in In less-developed settings, multiple generators may serve
all necropsy examinations. It may not always be possible to as both the primary and backup power sources. Scheduled
collect all of the recommended samples from each animal, monitoring and documentation of temperatures should
but the more consistently these goals can be achieved and be standard operating procedures to ensure consistent and
reports generated, the greater the chance that we will accu- continual storage temperatures. Whenever possible, samples
rately identify diseases and disease trends. Pathologists are should be immediately stored and frozen samples handled
often asked about the minimum set of tissues that should in ways that minimize freeze-thaw cycles.
be collected or that are necessary to make a diagnosis. Two significant sample management challenges are space
However, in situations in which collection of a complete and sample curation. With proper storage, tissues are a
set of tissues is not possible, a should be collected. This will valuable diagnostic resource for many years. Managing
vary by species and circumstance but should include those space, cost, power needs, ventilation, duration of research
tissues that are most likely to be most diagnostically relevant projects, chain of custody, and other aspects of short- and
(e.g., common sites of pathogenic infection). In general, long-term management of samples all need to be considered
these are the major organs and include heart, lung, liver, and strategies developed. Plans and protocols for the cura-
kidney, spleen, brain/spinal cord, stomach, and small and tion of sample storage and movement (e.g., for diagnostic
large intestine. Targeted tissue testing can also be performed testing, archiving, to fulfill research requests) must also
for specific suspect diseases (e.g., brain, lung, tracheobron- be developed and implemented. Sample information has
chial lymph nodes for suspected canine distemper testing; historically been maintained in paper logs; however, most
liver, kidney, fat, stomach content, spleen, hair, brain for people, even in remote locations, have access to computers
general toxicology screening; urine for domoic acid testing). and use electronic databases or software, which need to be
202 SE C T I O N 7 Diagnostics
TABLE
30.1 Diagnostic Tests and Recommended Sample Collection, Handling, and Storage
TABLE
30.1 Diagnostic Tests and Recommended Sample Collection, Handling, and Storage—cont’d
*This list includes several common test categories and general recommendations for sample collection, handling, and storage. Specific best practice protocols
should be consulted whenever possible, in development of your in-house standard operating procedures for sample management.
†
Recommended for labeling is pencil for glass slides, permanent marker or other appropriate inks for bags, tags, and labels that are resistant to alcohol, formalin,
other solvents, refrigeration, ultralow temperatures (e.g., −20°C, −80°C, liquid nitrogen) routinely used in necropsy or cytologic procedures or tissue storage/
handling/transport. It is also useful, in addition to labeling the container/bag, to place a labeled tag (labeled ink) in the container; cut pieces of Tyvek® may be used.
‡
Tissues collected for histology are typically placed in 10% neutral buffered formalin (NBF). For NBF and tissues placed in other fixatives, it is critical to maintain
a minimum ratio of 1 part tissue to 10 parts fixative to ensure proper fixation. Salinity and pH of fixative are important considerations for marine invertebrates. For
invertebrates, 10% formalin or sea-water formalin are adequate fixatives in most cases. Specialized fixatives are preferred for some species such as zinc-formalin
(Z-fix) for corals and Davidson solution for soft-bodied invertebrates (cnidaria). Decalcification can cause significant artifactual changes in the heavily mineralized
tissues of some species, particularly corals and echinoderms. If the structure of the skeleton of these animals is critical for evaluation, the pathologist should
consider submitting tissues for processing in a mineralized state (such as through plastic embedded and sectioning at a bone pathology laboratory) or a slow
method of decalcification should be chosen (using the acetic acid in Davidson solution or pH-balanced EDTA). Enrobing the delicate tissues of coral species in
alginate prior to decalcification has proven an excellent method for protecting the delicate surface polyps while retaining skeletal structural features. Formic acid
and hydrochloric acid may be used for decalcification, but tissue disruption through gas bubble formation should be recognized and not interpreted as a lesion.
For species with a chitin-based ectoskeleton, Pyreni solution may be used to soften the chitin and provide soft supple tissue for sectioning.
§
Recipes for marine invertebrate specific electron microscopy fixative are recommended.
NBF, Neutral buffered formalin; VTM, viral transport medium.
204 SE C T I O N 7 Diagnostics
regularly updated and backed up to prevent catastrophic platforms in the not too distant future; robust systems for
data loss. In some situations, for example, large active sample secure data storage and backups will therefore continue
repositories or zoo collections with large sample inventories, to be a necessity. These databases contain a treasure trove
dedicated staff may be needed to curate and manage sample of baseline and disease-related information and available
acquisition and disposition information and databases, as samples, all of which are hugely valuable in retrospective
well as sample retrieval and shipment. Archived inventories and prospective projects focused on wildlife ecology, health
should be reviewed every 6 months to update additions and monitoring and disease surveillance, and conservation
remove tissues that become redundant. projects in zoos and other managed and free-range settings.
Most facilities do not have the in-house resources or Shared access to these valuable resources should be the
expertise to perform the broad range of tests that are rule rather than the exception (e.g., collaborative research
run as components of a complete necropsy/postmortem projects, biomaterials requests, contributions to SSP/EEP
examination. Shipment of samples to external laboratories and TAG programs). Whether retrospective or prospective,
is therefore a requisite of many necropsies. All sample ship- inclusion and participation of wildlife veterinary patholo-
ments must comply with all local, state/provincial, federal/ gists, who can appropriately interpret and contextualize
national, and international laws and regulations to ensure data to prevent misinterpretation (a common problem
human and animal safety and sample viability. Special in data review by nonpathologists) and contribute to and
packaging, handling, and labeling may be required (e.g., guide the development of protocols that involve necropsy
for shipments containing dry ice or LN2). For shipment and sample collection from free-ranging wildlife or zoo
to local laboratories, contacting the lab to discuss specific animals, are essential.
package labeling, submission forms, test requests, and
minimum sample volumes is of value. For international Communication
shipments, export and import permits to ship samples and
compliance with treaties and regulations concerned with Communication during a necropsy or active mortality
the international movement of samples from endangered or investigation and after are critical, both practically and politi-
managed species (e.g., Convention on International Trade cally. Communications should be directly coordinated first
in Endangered Species of Wild Fauna and Flora [CITES], with managers and/or managing authorities (institutional,
US National Oceanic and Atmospheric Administration local, national, etc.) before being more widely disseminated.
[NOAA for marine mammals], CDC [primates, rodents, Information is generally of interest and should be shared
civets, bats]) may also be required (see Chapter 4). The regularly and transparently with stakeholders (e.g., clini-
regulations and paperwork differ in every country and can cal veterinary staff, keepers, curatorial staff, collaborators,
be quite complicated, and processing can be excruciatingly indigenous farmers, public) as results become available and
time consuming. However, it is essential to understand all interpretations and conclusions are drawn. Immediate com-
steps in the process, be in compliance, and pay attention munication of test results is also specifically important for
to all of the many details. Failure to do so knowingly or medical treatment and/or mitigation of disease in contact
unknowingly may result in significant fines, import/export with wild or domestic animals, staff, or others (e.g., public
permit refusal, confiscation and return of samples to their health networks) and for directing additional ancillary diag-
country of origin, or confiscation and destruction of samples nostic testing. As mentioned previously, suspicion or confir-
by border patrol agents. Because of the latter, it is often a mation of listed or reportable diseases must be immediately
good idea, whenever possible, to maintain a duplicate set shared with appropriate local, national, and international
of samples at your facility as an assurance against problems authorities (see above), who may need to participate in
that might occur during the shipping process. In addition, disease management. The format for the communication
many diagnostic labs will discard tissues, glass slides, or varies depending on need and may include sharing of
paraffin blocks after a certain retention time, and many complete or summary pathology reports; peer-reviewed
claim ownership of all submitted specimens (which they publications; presentations at professional conferences or
may or may not have the legal authority to claim). Written community meetings; development or iterative revision of
agreements with laboratories may be required to ensure protocols, practical technical manuals, and other teaching
return, retention, or ownership of your samples. materials; or other forms of communication with the media,
including interviews and articles in the lay press. For high-
Data Management and Sharing profile cases or outbreaks with high public interest, it may
be of value to identify a communications lead or team to
All historical data, pathology reports, ancillary diagnostic field questions and provide comments that are informed by
test results, photographic documentation, and sample the pathologist and necropsy team. Creating a list of key
archiving and curation must be organized and managed. talking points to be used by the communications team is
In many settings the records exist as/in a combination of beneficial to guide messaging, especially when conveying
paper and electronic documents and databases. However, complex scientific information to the public. Care should
technologic advances will likely push all activities related to also be taken to communicate only confirmed data or to
documentation, even that done in the field, onto electronic qualify all results as preliminary until they are confirmed.
CHAPTER 30 Wildlife Necropsy Primer 205
Introduction the surface of the lung, and gas within the gastrointestinal
tract. Modern CT is also best suited to accommodate large
The value of cross-sectional imaging (computed tomog- patient sizes because CT x-ray photons are less attenuated
raphy [CT] and magnetic resonance imaging [MRI]) in by large patient tissues than ultrasound waves, resulting
zoological medicine was quickly recognized following the in less deterioration of image quality while maintaining
first emergence of the technology in the 1970s, but wide- excellent spatial resolution.
spread application was restricted due to cost and equipment The superiority of CT over radiographs and ultrasound
availability.1 Currently, CT and MRI scanners are installed for detection and characterization of innumerable condi-
in veterinary facilities around the globe, providing many tions is well established in domestic animals. For instance,
zoos with ready access. As the technology continues to the diagnostic value of CT in tumor and metastasis detec-
rapidly evolve, imaging once reserved only for complex, tion is broadly recognized,6 and CT broadly allows for
high-profile cases has now become standard, routine care characterization of orthopedic lesions that are otherwise
in some zoological facilities. undetectable. It also allows superior assessment of soft tissue
Entire textbooks are devoted to the veterinary applica- and bony structures in the sinuses, pulmonary parenchyma,
tions of CT and MRI, and the reader is directed to these abdominal viscera, vasculature, and lymph nodes, among
sources, as well as theoretical medical imaging texts, for many others. Studies specifically characterizing the superi-
discussion of the underlying physics and principles of image ority of CT diagnostic sensitivity in nondomestic animals
acquisition, image interpretation, minimization of artifacts, are limited, but extrapolation is valid. In birds, CT has
and advanced applications.2–5 Technologic evolution is been shown to be superior to radiographs for detection of
rapid, and readers are also referred to other sources to best the early stages of upper airway disease and other soft tissue
assess current equipment options. The aim of this chapter disorders of the head.7 Radiographic evaluation of birds
is to provide an overview of the value of cross-sectional in many zoos is performed on a regular basis as a screen-
imaging in nondomestic species and the optimization of ing tool for aspergillosis, but mild abnormalities may be
image acquisition in a zoo setting. Discussion is heavily detectable only with CT.8,9 Similarly, pulmonary parenchy-
focused on CT because its use in zoological medicine is far mal lesions in cetaceans that are silent on both ultrasono-
more widespread (see also Chapter 32). graphic and radiographic evaluation can be detectable on
CT (Fig. 31.1).10,11
Radiographs and abdominal ultrasound are commonly
Benefits of Cross-sectional Imaging in performed in zoos as part of preventative medical examina-
Zoological Medicine tions, and an additive integration of CT into such routine
practice offers many benefits for preventative care. For
Cross-sectional imaging is by definition superior to example, traditional three-view radiographs in great apes
radiography due to the elimination of superimposition present many positioning challenges under anesthesia. Their
artifact (i.e., three-dimensional [3D] data of a 3D patient, size results in marginal diagnostic quality for fine scale
instead of 2D data of a 3D patient). It is not susceptible to lesion assessment and often requires hemithoracic views.
critical ultrasonography artifacts such as shadowing, which In contrast, CT imaging provides a comprehensive study
obliterates an operator’s ability to see tissues deep to bone, that allows detection of pulmonary lesions undetectable
206
CHAPTER 31 Use of Computed Tomography/Magnetic Resonance Imaging in Zoological Medicine 207
A C
B D
• Figure 31.1 Computed tomography (CT) provides superior detection of many diseases in zoo species.
Shown are a lateral thoracic radiograph (A) and a parasagittal CT reconstruction (B) of a bottlenose
dolphin. Note the CT projection reveals sites of peribronchiolar consolidation (arrows) not detectable
radiographically. Also shown is a coronal reconstruction (C) demonstrating central peribronchiolar lesions
and foci of ground-glass opacification (arrows) not visible on ultrasonographic evaluation of the normal
peripheral lung (D).
radiographically (Fig. 31.2), superior detection of sinusitis has proven valuable for objective body condition assessment
and air sacculitis,12 and complete assessment of degenerative and establishment of weight goals by monitoring adipose
changes in spinal and thoracic joints.13 stores (Fig. 31.3).
In many species, anatomic structures limit the value Cross-sectional imaging allows unparalleled assessment
of other diagnostic imaging modalities. Protective features of dentition and is of particular value in species with a
(e.g., plates, shells, scales, quills) are superimposed over narrow oral gape, such as aardvarks (Orycteropus afer) and
internal structures on radiographic images, drastically macropods. Integration of CT into examinations in these
decreasing diagnostic utility. These same features can species allows for better preventative care through earlier
also render ultrasonography impractical. Cross-sectional detection of periodontal disease and superior monitoring
modalities are generally not impeded by these features, and of response to treatment.15 CT provides unobstructed
they can be removed from view in a digital environment. views of the jaw and tooth roots to allow lesion localiza-
The use of CT is widely recognized as a best practice for tion (Fig. 31.4) and precise serial monitoring. Integration
imaging chelonians and respiratory disease in reptiles.9,14 of CT into preventative healthcare for such species has
CT provides great value in assessing general health of species been greatly beneficial. The authors have also found value
with anatomy that precludes easy palpation. For instance, it in using CT to localize foramina for dental nerve blocks
208 SE C T I O N 7 Diagnostics
Equipment Considerations
Equipment Access
In the past, access to CT and MRI equipment posed a
challenge to many zoos. Currently, arrangements with
veterinary referral clinics, veterinary colleges, and human
imaging centers often provide zoo clinicians with access to
cross-sectional imaging, but routine use at an off-site facility
B remains challenging. Dangerous and large animals must be
maintained under anesthesia during transport, increasing
anesthetic risk for the patient due to prolonged procedure
time and inability to maintain an ideal anesthetic environ-
ment in a moving vehicle. Human safety considerations also
exist when transporting dangerous zoo animals off-site into
less-controlled environments. Logistical problems may exist
when maneuvering large animals through facilities designed
for small animals or people, and scheduling considerations
often necessitate “after hours” use. Even when animals
can be placed in a crate and anesthetized off-site, having
C access to appropriate emergency equipment and staff can
be challenging. Bringing zoo animals into a veterinary facil-
• Figure 31.2 Computed tomography (CT) provides superior diag- ity in proximate contact with companion animals creates
nostic capabilities in preventative medical care. Axial MinIP (minimum infectious disease risks. Use of human facilities leads to
intensity projection) (A) and coronal CT reconstruction (B) from a pre-
ventative medical examination on a gorilla reveal external compression
both veterinary concerns of anthropozoonoses and liability
and narrowing of the left mainstem bronchus (curved black arrows) and concerns related to zoonotic diseases. Public relations and
hypoinflation of the left lung. These signify important considerations for privacy considerations must be considered before transport-
patient positioning and anesthesia. A ventrodorsal radiograph (C) fails ing zoo animals, and the expenses associated with facility
to diagnose bronchial constriction. Radiographs also require acquisi- use and transportation may be considerable.
tion of multiple hemithoracic views due to patient size.
The immense value of on-site equipment at a zoo cannot
be understated. Safety and collection health considerations
and for measuring canal depth in endodontic repairs are dramatically decreased. Time under anesthesia is mini-
(see Fig. 31.4). mized to the extent that a full body multidetector CT scan
The large size of many zoo species can restrict imaging can be performed more quickly than a radiographic study.
success with ultrasound (limited penetration capabil- New concerns noted during an exam can be addressed
ity) and radiography (practical and technologic limits). immediately with advanced imaging without the need for
Advancements in human bariatric medicine have resulted an additional anesthetic event. Complete imaging studies
in greater availability of CT scanners with a large gantry can be performed during preventative healthcare exams,
bore (≥90 cm) and robust table design (≥300 kg) that offer increasing early detection rates. Considerations of cost
immense versatility for large zoo species (Fig. 31.5). per case are eliminated, encouraging frequent, routine use
Three-dimensional renderings created from cross- that results in higher-quality medicine and a new standard
sectional data provide further value in assessing species that of care. The controlled zoo environment also facilitates
lack thorough anatomic descriptions and medical illustra- the ability to perform interventional CT procedures that
tions. The manipulation of 3D images in virtual space is enhance diagnostics and treatment. In-house equipment
CHAPTER 31 Use of Computed Tomography/Magnetic Resonance Imaging in Zoological Medicine 209
A B
C
• Figure 31.3 Computed tomography (CT) provides versatility in imaging patients with protective outer
coverings. A ventrodorsal radiograph (A) of a short-beaked echidna provides limited diagnostic information
due to superimposition of quills. A coronal soft tissue CT reconstruction (B) of the same animal reveals
superior detail. Body condition can also be evaluated using CT to quantify adipose stores (shaded blue);
this provides more accurate characterization than radiographs or visual examination (represented by a
three-dimensional reconstruction of the animal [C]).
allows clinicians to easily maintain advanced anesthetic upgrades to newer CT scanners incur relatively little instal-
monitoring equipment, intravenous access, patient ther- lation cost. As technology advances, refurbished equipment
moregulatory support, and ventilatory support (Fig. 31.7). is broadly available at steeply reduced prices. Many zoos
also have broad community support, which may assist in
Selection of Computed securing a CT scanner from a local hospital upgrading to
Tomography Equipment a newer unit.
Preventative maintenance and repairs represent consider-
The investment required for a scanner is substantial and able ongoing costs. A service contract can help to keep these
remains the principal limitation to the widespread applica- annual costs consistent and avoids unexpected fluctuation
tion of CT in zoological medicine. Installation into an in operating budgets. Facilities must also be willing to invest
existing facility may require upgrades to electrical utilities, appropriately in staff training to ensure proficiency in CT
surge suppression equipment, ventilation modifications for operation and rudimentary image assessment. Human CT
adequate cooling, and installation of lead-lined drywall and equipment is generally operated by registered CT techni-
windows. Special care should be taken to ensure adequate cians, but most veterinary facilities use veterinary techni-
floor space is available for maneuvering gurneys with large cians with advanced training. Veterinarians must become
patients, anesthesia equipment, and interventional supplies. familiar enough with CT principles to articulate their study
If building infrastructure is designed appropriately, future goals and corresponding scan parameters.
210 SE C T I O N 7 Diagnostics
B C
D E
• Figure 31.4 Dental evaluation with computed tomography (CT). Three-dimensional (3D) (A), axial
(B), and parasagittal (C) CT images of a western grey kangaroo illustrate the value of CT for evaluating
dentition in species with a narrow oral gape. Note the severe focal lysis of root apices and alveolar bone
of this right maxillary molar (arrows in B, C). A normal unerupted molar is also seen (*). Sagittal (D) and
3D (E) CT reconstructions of the teeth of an Amur leopard illustrate the value of CT in efficient endodontic
repair in large animals by providing precise pulp cavity measurements and foraminal locations for nerve
blocks (arrow).
Conventional (fan-beam) multidetector CT operates by equine veterinary medicine over the past decade, driven
simultaneously scanning multiple slices, thereby improv- by its lower cost and installation ease. CBCT operates
ing image quality (spatial resolution) and reducing scan on standard electrical supply and requires fewer safety
time. Scanners are routinely referred to by their number considerations due to lower radiation output. Although
of detector arrays (e.g., 64-slice CT) and the smallest superior to traditional radiography with regards to lack
achievable slice thickness (e.g., 0.5 mm). Dual-source CT of superimposition, CBCT has numerous drawbacks
is a generational technology step that combines two x-ray compared with conventional CT. CBCT studies are
sources and detectors that rotate simultaneously to further more sensitive to patient motion, have poor soft tissue
improve resolution and speed, while decreasing radiation contrast resolution, and are prone to windmill streak-
dose. Such advanced machines (>320 slice, 0.3 mm) allow ing artifact (Fig. 31.8).16,17 Given the intended use in
imaging of the entire human heart within one beat and human medicine for small structures of the head, these
other specialized applications. Although not yet broadly artifacts are amplified in larger veterinary patients.
available in veterinary medicine, such technology will Furthermore, tissue attenuation cannot be quantified
become increasingly accessible to zoos over time. reliably in Hounsfield units with CBCT, leading to sig-
Commonly used in human maxillofacial imaging, nificant restrictions in diagnostic interpretation relative to
cone beam CT (CBCT) has emerged in small animal and conventional CT.18
A B
D E
• Figure 31.5 Bariatric computed tomography (CT) scanners offer enhanced diagnostic capabilities for large zoo species. (A) Pygmy hippo CT
scan. (B) Okapi pelvic CT scan. (C) Sagittal soft tissue CT reconstruction of a sloth bear. Three-dimensional CT volume renderings of a sloth bear
(D) and an Amur leopard (post contrast) (E).
212 SE C T I O N 7 Diagnostics
A B
• Figure 31.6 Thoracic three-dimensional volume renderings of a bottlenose dolphin; dorsal view. (A)
Lung surface. (B) Bronchial tree with digital excision of pulmonary tissue. The CT data enable virtual
endoscopy in a digital environment to create an efficient endoscopic route to a precise location in the
live animal for sample collection. ([A], Ivančić M, Solano M, Smith CR: Computed tomography and
cross-sectional anatomy of the thorax of the liver bottlenose dolphin (Tursiops truncatus). Anat Rec
297:901–915, 2014.)
A B
C D
F
• Figure 31.8 Cone beam computed tomography (CBCT) compared with conventional CT (CCT) in large zoo species. (A) Axial CBCT image of
the lungs of an American alligator. (B) Axial CCT image of the lungs of an Orinoco crocodile. (C, E) Axial and sagittal CBCT reconstructions of a
giant anteater in lung and soft tissue reconstruction algorithms. (D, F) Axial and sagittal CCT reconstructions of a giant anteater in comparable
reconstruction algorithms. Note that CBCT provides a limited field of view and poor soft tissue contrast resolution compared with CCT and is prone
to motion and windmill streaking artifacts (visible as radiating and semicircular lines in [C], and horizontal lines in [E]).
Such software should at minimum allow for the genera- determining resolution. A tightly collimated DFOV thus
tion of multiplanar reformats (sagittal and coronal planes), provides optimal resolution. Retroactive reconstruction of
minimum/maximum intensity projections, synced review raw data can be performed during postprocessing to address
of multiple axial series, and 3D reconstruction. this issue, allowing enhanced DFOV resolution within a
Expertise in image interpretation is critical to proper wider original SFOV.
diagnosis. As medical knowledge continues to grow expo- Placement of patient limbs is important to maximize
nentially, veterinarians cannot be experts in all fields. In image quality of specific organs. To decrease tissue pen-
much the same way most clinical veterinarians are not etration depth by the beam, forelimbs generally should be
adequately trained to evaluate histopathology, most veteri- extended cranially for imaging of the thorax or the limbs
narians are not adequately trained to form detailed diagno- themselves and positioned caudally for imaging of the head
ses from cross-sectional imaging. The diagnostic knowledge or neck. Orthopedic implants should be positioned away
and expertise a veterinary radiologist provides cannot be from the ROI to reduce streaking from scatter, beam hard-
understated. A radiologist’s expertise is reliant on familiarity ening, and edge effects. Patient recumbency can influence
with the anatomy of a given species. Medical radiologists the degree of motion artifact that occurs with respiration.
may be of great value in interpretation of images from Positioning has been shown to also significantly affect
an ape, but their expertise diminishes in other taxonomic lung volume in some species (e.g., red-eared slider turtles
clades. Veterinary radiologists possess a wider breadth of [Trachemys scripta] and Humboldt penguins [Spheniscus
expertise, particularly in species with a close domestic humboldti]).19,20
animal counterpart. Normal anatomic structures, such as Appropriate anesthetic management during a scan is
the avian glycogen body, can be easily misinterpreted as paramount. Minimally, an ECG and capnograph should be
pathologic lesions by radiologists unfamiliar with a given visible from outside the CT room during the scan. Monitor-
species. Radiologists with specialized expertise in nondo- ing equipment may be left in place during scout scans
mestics are highly desirable within zoological medicine and used for study planning, but any wires or metal equipment
provide tremendous clinical guidance. that passes through the gantry should be removed prior
to full diagnostic scans, to avoid artifacts. Intubation and
Optimization of Computed Tomography ventilator use are recommended for long, complex scans.
Patient temperature should be monitored closely because
for Zoological Applications CT rooms are cooled to maintain the equipment. Small
Procedure Planning patients can become hypothermic quickly, and patient
warmers should be used proactively.
Advanced imaging technology necessitates a degree of
technical knowledge and prescan planning far more Scan Settings
complex than radiography or ultrasonography. The added
logistical challenges posed by anesthetized zoo patients Consultation with a radiologist is recommended prior to
demand maximal preparation, coordination, efficiency, and any CT scan to ensure image acquisition settings are set
expertise. By having practical knowledge of CT acquisition appropriately. Slice thickness, interval, pitch, rotation speed,
parameters, clinicians can greatly enhance the diagnostic and other settings all impact the number of axial images
quality of a study. A mutual understanding of the primary acquired during a scan. This, combined with the length of
clinical concern by both the clinician and radiologist is the ROI, determines the scan duration. Acceptable scan
critically important for appropriate planning. duration varies based on the species, health status, anesthetic
considerations, and other factors. Contrast enhancement
Patient Positioning and Anesthesia adds additional time. In large patients, high tube current
may necessitate a wait period between scans to allow for tube
With large patients, positioning on the CT table is para- cooling. It is important that clinicians and CT operators
mount to ensure unimpeded movement through the gantry. communicate closely regarding acceptable scan duration.
Weight limits on the cantilevered portion of the table may Clinicians should have a general understanding of recon-
necessitate splitting a study and craniocaudally rotating the struction algorithms. Images should be acquired using the
patient between scans. In such instances, scanned regions proper algorithm (e.g., soft tissue, bone, lung) for a given
should overlap to avoid data loss. The anatomic region ROI. For instance, lung tissue should not be evaluated
of interest (ROI) should be horizontally and vertically using a soft tissue algorithm. Acquisition with the wrong
centered in the scan field of view (SFOV), but vertical algorithm severely limits diagnostic quality because window
centering may not be possible in large patients, depending width and level adjustments during viewing can only
on bore size. In zoo species with limited reference data, the partially compensate for an incorrect algorithm. Multiple
use of a wide SFOV avoids cropping of anatomy that may reconstruction algorithms can be applied simultaneously
be of diagnostic or future reference value but can adversely during scanning or retrospectively reconstructed from
impact image resolution. The DFOV for reconstructed raw data. As a general guide, scans in zoo species are best
images is spread across a fixed matrix (generally 5122 pixels), acquired using multiple algorithms.
CHAPTER 31 Use of Computed Tomography/Magnetic Resonance Imaging in Zoological Medicine 215
• Figure 31.9 Postprocessing of raw computed tomography (CT) data. Shown are original (left) and
postprocessed (right) axial CT reconstructions from a gorilla thorax at the level of the shoulder. Note that
reducing the display field of view and application of an adaptive noise reduction algorithm to the raw CT
data greatly decrease image noise, increase resolution, and improve image quality in large zoo species.
Collecting CT data beyond the specific region of clinical slower CT scanners. Respiratory gaiting eliminates this
concern is valuable, given the relative lack of reference images artifact by syncing image acquisition with breathing but
in many species. Modern scanners allow for efficient acqui- necessitates the use of a ventilator in veterinary patients
sition of whole body studies. However, clinicians should that do not breathe on cue. Alternatively, decreasing slice
remain cognizant of dose efficiency (achieving diagnostic thickness, increasing slice overlap, and adjusting rotation
image quality at the lowest possible dose), particularly in speed may minimize the impact of individual breaths.
young animals and long-lived species exposed to repeated Recognition of significant motion artifact during a scan
scans. may necessitate image reacquisition.
Streaking artifacts can occur in highly attenuating skel-
Contrast Enhancement etal regions (shoulders and pelvis) due to photon starvation
(i.e., an insufficient number of photons reach the detector).
Pre- and post-contrast imaging is routine in veterinary Proper patient positioning can decrease this artifact. Inad-
CT studies, unless scan duration is a concern or there are equate dose can also result in increased noise and dimin-
contraindications to intravenous contrast administration ished image quality. Automatic mA adjustment in modern
(e.g., patient dehydration).21 There is still much to be scanners minimizes artifacts by increasing tube current for
learned about contrast use in zoo species due to variations highly attenuating sections within a ROI. Photon starva-
in anatomy and physiology. Nonionic, iodinated contrast tion is a notable challenge in large species because further
agents (e.g., iohexol) are safer for young, geriatric, and increases in tube current may not be possible. In such cases,
debilitated patients than ionic solutions (e.g., iothalamate, postprocessing of raw data with adaptive noise reduction
diatrizoate). Timing of image acquisition is critical for algorithms can greatly enhance image quality (Fig. 31.9).
meaningful angiography, excretory imaging studies, and Given the rise in bariatric imaging and emphasis on dose
assessing tissue enhancement. Large-bore catheters facilitate efficiency in humans, future technology will likely continue
the necessary rapid administration of contrast, but a power to aid image quality for zoo species.
injector is beneficial in large species. Iodinated contrast can
also be administered enterally for luminal gastrointestinal
enhancement. Interventional Computed
Tomography Applications
Artifact Detection and Prevention
Interventional radiology refers to the use of imaging
Imaging artifacts can severely impact the quality of a scan procedures to guide minimally invasive diagnostic and
by depicting a structure on an image that is not present therapeutic techniques. Software features on CT scanners
within the patient. Detailed discussion of artifact formation facilitate the collection of precise biopsies (full-core and
is available in other sources.2,5,16,22 Recognition of some aspirates). CT-assisted biopsies can be obtained using laser
common artifacts (e.g., beam hardening, ring detector, guidance, trajectory assessment, and repeated short-range
partial volume, metal streaking) can be partially corrected scans. CT-fluoroscopy in modern scanners allows for real-
with software applications or recalibration. time visualization of needle placement and advancement.
Motion artifact from respiration can significantly affect Using this technology, highly precise procedures in delicate
thoracic imaging in zoo animals, particularly when using anatomic regions are possible (Fig. 31.10).
216 SE C T I O N 7 Diagnostics
A B
C D
E F
• Figure 31.10 Interventional computed tomography (CT). A coronal CT reconstruction (A) reveals soft
tissue attenuating material filling the left maxillary sinus (white arrow) in a white-cheeked gibbon, not
apparent radiographically (B). On-site CT equipment at a zoo facilitates interventional CT procedures (C).
Using CT laser guidance, detailed measurements, and focal repeated scans (D), a Rosenthal needle is
precisely directed into the sinus to obtain diagnostic aspirates while avoiding delicate adjacent anatomic
structures (E). A three-dimensional volume rendering illustrates needle placement (F).
CHAPTER 31 Use of Computed Tomography/Magnetic Resonance Imaging in Zoological Medicine 217
218
CHAPTER 32 Moving Beyond Survey Radiographs 219
A B
• Figure 32.1 (A) and (B) African elephant (Loxodonta africana) lateral digital radiology (DR) image of
necropsy specimens along the dorsal margin of the proximal and distal inter-tarsal joints of the left limb,
with mild to moderate osteophyte formation (black arrows) present consistent with osteoarthrosis and
degenerative joint disease (Fig. 32.1A). Computed tomography image of the left tarsus, seen in Fig.
32.1A, of the African elephant showing severe erosions and fragmentation of the articular margins and
subchondral bone is present (white arrow), which was not apparent in the DR images (Fig. 32.1B). (Images
courtesy of Ajay Sharma, University of Georgia College of Veterinary Medicine.)
A B
• Figure 32.2 (A) and (B) 3D reconstruction from computed tomography (CT) of the dorsolateral view
of the right hind foot of an Asian elephant (Elephas maximus), showing a bipartite distal phalanx (white
star) and absence of the middle phalanx of digit 5 (white arrow) (Fig. 32.2A). Sagittal slice CT image of an
Asian elephant, showing remodeling of the metacarpal (black arrow) and fracture of the middle phalanx
digit 4 of the right hind foot (black star) (Fig. 32.2B). (Courtesy PeerJ, Inc 2017. Regnault, et al.)
allows for built-in controlled comparison between abnor- The purpose of two orthogonal images is to visualize
malities localized to a specific region. This is then based the region(s) of interest in two different projections and
on physical examination, observation, and/or history, and determine how the abnormality silhouettes on these radio-
compared to those without mirror-image structures that are graphic views. Because this compresses a three-dimensional
of normal appearance. For this comparison to occur within structure into a two-dimensional image, we use the second
the given patient or within a given species, it is important radiograph to ascertain the location of an object within the
that positioning be performed in a consistent and reproduc- third plane due to orthogonal nature. Some changes are
ible manner for all regions of interest. There are standard only apparent on a single view due to superimposition or
views used to complete survey radiographic examination absence of radiographic changes within the opposite image
of normal structures with all requiring a minimum of two from the beam not striking the interface at a tangent; thus
orthogonal images (taken at 90 degrees to one another) to not demonstrating attenuation of the beam in the projected
complete the study.12,26–30 image (no changes in radiographic signs are apparent on
220 SE C T I O N 7 Diagnostics
A B
• Figure 32.4 (A) and (B) Computed tomography (CT) of the right carpus of Asian elephant (Elephas
maximus). CT cross section (Fig. 32.4A) through the proximal and distal carpus (Fig. 32.1A and 32.2), and
3D reconstruction (Fig. 32.4B), showing the obvious presence (arrows) of osteoarthrosis and degenerative
change in the metacarpus and carpus. (Images by Rush EM.)
CHAPTER 32 Moving Beyond Survey Radiographs 221
A B
• Figure 32.5 (A) and (B) Orthogonal skull digital radiology of a juvenile badger (Taxidea taxus) with
neurologic signs and chronic sinusitis and rhinitis. (Courtesy Antech Imaging Services, 2017.)
A B
• Figure 32.6 (A) and (B) Computed tomography image with multiplanar reconstruction of the badger
(Taxidea taxus) skull, ventrodorsal (Fig. 32.6A) and lateral (Fig. 32.6B) views. Note the deviation of the
rostral sinuses from the midline. Infiltrative change in the sinuses is designated by the arrows. The green
crosshairs indicates the location in the skull with respect to sections taken in Fig. 32.7. (Courtesy Antech
Imaging Services, 2017.)
1. DR images may be reviewed immediately after exposure format (DICOM) for transfer to digital database.17,29
on a local monitor to determine if views are correctly Analog films must be digitized with a scanner or possibly
positioned and include the entire region of interest. Both a camera before they are available on a DICOM network.
analog radiographs and computed radiology (CR) require Each time an image is copied into an alternate format,
a processing step before an image may be observed. The some of the information and detail will be lost. Often
patient cannot be moved from the location of image this will produce an acceptable image, which retains
capture (most often a specified room or table) before the diagnostic detail; however, other times it may reduce the
study is completed. DR allows for decreased anesthesia copied image to nondiagnostic with regard to quality
time required to obtain images in comparison to analog and detail.17
and CR. DR removes the processing step and provides an
instant view of the produced image for quick evaluation Imaging Paradigm—What Next
of technique and positioning.
2. Images produced by DR and CR are typically produced After radiographs of the region of interest have been
in Digital Imaging and Communications in Medicine obtained and reviewed for abnormalities, consideration for
222 SE C T I O N 7 Diagnostics
A B
• Figure 32.7 (A) and (B) Multi-planar reconstructed computed tomography of the badger (Taxidea taxus)
skull, axial projection showing the severe destructive and infiltrative changes in the maxillary sinuses at
different levels of the sinus (arrows). (Courtesy Antech Imaging Services, 2017.)
A B
• Figure 32.8 (A) and (B) Orthogonal digital radiology projections of a red-tailed hawk (Buteo jamaicensis)
skull with a large growing fracture not visible on radiographs. Note the soft tissue enlargement and poor
definition of the lesion (arrows). (Courtesy JZWM, Rush EM, Shores A, Meintel S, et al: Growing skull
fracture in a red-tailed hawk (Buteo jamaicensis). J Zoo Wildl Med 45(3):658–663, 2014.)
additional noninvasive diagnostic imaging may be needed center of the CT gantry without rotation or bending of the
to further evaluate an area or organ system. If detail is poor skull. The hard palate of the skull should be parallel to the
or absent in the abdomen, then progression to an alternate scan table to allow direct acquisition of true axial slices of
imaging modality (e.g., CT or US) may be needed to see the the skull and brain.
soft tissues within the abdomen that are surrounded by fluid Slices generated by the CT machine should be accom-
or have loss of regional adipose tissue. Ultrasound is ideal panied by scout views (similar to radiograph and produced
for focal examinations but may be difficult for evaluation in two orthogonal planes) and used in planning the slice
of an entire abdomen due to superimposed gas, underly- acquisition with regard to start and finish of axial slices.
ing pain, and/or pathology, or the massive size of a lesion The scout views are linked with the axial slices at time of
that is contiguous with multiple organs, limiting ability review on the computer monitor so that relative position
to determine site of origin. If US is nondiagnostic, then of the slice within the thorax, abdomen, or extremity is
whole body CT examination to image the entire thorax and readily apparent by superimposition of the slice on the scout
abdomen may be warranted.14,17,20–26 image. With these, it may be difficult to determine which
Just as in radiography, movement by the patient during ribs and vertebrae are present within a given axial slice when
CT image acquisition decreases image detail and could reviewing axial images.
mask pathology. Patient positioning is critical to allow The major advantage of CT images is production of slices
obtainment of CT axial images in the standard imaging of body tissue (determined by slice thickness set by operator
plane for review. The patient must be positioned in the before scan) with removal of superimposition artifact, and
CHAPTER 32 Moving Beyond Survey Radiographs 223
A B
• Figure 32.9 (A) and (B) Cross sectional computed tomography (CT) image made prior to surgery of the
red-tailed hawk (Buteo jamaicensis) demonstrating the defect in the skull with slight soft tissue protrusion
and mild cerebral herniation. This image also demonstrated the smooth (chronic) nature of the edges of the
skull and the site of the defect which is notated with an arrow (Fig. 32.9A). Sagittal and 3D reconstruction
images of preoperative CT scan clearly define the skull defect with the smooth, lipped edges that were
noted in surgery. The arrow clearly notes the defect in the skull (Fig. 32.9B). (Courtesy JZWM, Rush EM,
Shores A, Meintel S, et al: Growing skull fracture in a red-tailed hawk (Buteo jamaicensis). J Zoo Wildl
Med 45(3):658–663, 2014.)
increased ability to differentiate tissues and organs due to or to guide surgery approach and resection/debulking
differential attenuation of the x-ray beam within the body options.17,29,34
(detected by the CT during image acquisition).
Another advantage of CT is multiplanar reformatting. Summary of Imaging Planning
This is where cross-sectional images from a region of inter-
est are reconstructed in the other two orthogonal planes: 1. Standardized positioning of anatomic region for all
sagittal and coronal (sometimes referred to as dorsal plane). species.
In order to produce high-resolution reformatted images in 2. Minimum of two views to complete the radiographic
the desired plane of interest, thinner axial slices are needed study.
and patient motion must be minimized.17,29,34 3. Sedation/anesthesia to minimize movement and control
Contrast-enhanced images should be obtained following respiration.
standard noncontrast enhanced CT imaging. This increases 4. Transfer images to data storage center for storage and
conspicuity of vessels, regions of increased blood flow, or possible review by specific radiologist and/or clinicians.
abnormal vasculature permeability that results in regional Referral of images to an imaging expert may help to
contrast enhancement. This enhancement is often needed interpret the diagnostic images and help set up protocol
to determine size and location of lesions observed during for further assessment of the lesion via additional imaging
a scan. Some lesions may not be visible, or size may be modalities. This relationship may be very productive and
underestimated/overestimated due to contrast enhancement. advantageous as radiologists are trained to consider a next
Once CT images have been obtained, interpretation of imaging option or step in the continued evaluation of
the images should produce a reasonable list of differential patients, based upon findings of initial survey radiographic
diagnoses based on radiographic findings, clinical history, images.
and possible signalment of patient (species, age, sex,
geographic location). The location of the lesion as well as Future Directions
the effect on adjacent structures (including invasion, dis-
placement, or encapsulation/encasement) may be used for Both zoological and radiographic knowledge of multiple
planning the next course of action. Changes seen are often species could significantly increase over time with a shared
nonspecific, and the CT machine may be used to guide database. The ability to predict which possible disease
aspiration and biopsy of lesions for pathologic assessment, processes are present based on location, appearance, and
224 SE C T I O N 7 Diagnostics
Acknowledgments
The authors would like to thank Paul Fisher and Antech
Imaging Services (AIS), for their support and dedication
to advancement of imaging diagnostics, including offering
development of an accessible database for zoological species
with AIS. Thanks to Dr. Sophie Regnault for images.
2. Fowler ME: Foot disorders. In Fowler ME, Mikota SK, editors: of noncardiac thoracic disease in the dog and cat, Vet Radiol
Biology, medicine and surgery of elephants, Ames, IA, 2006, Ultrasound 46:114–121, 2005.
Blackwell Publishing Professional, pp 271–290. 20. Steinmetz HW, Zimmerman N: Computed tomography for the
3. Gage L: Antemortem diagnostics. In Fowler ME, Mikota SK, diagnosis of sinusitis and air sacculitis in orangutans. In Fowler
editors: Biology, medicine and surgery of elephants, Ames, IA, ME, Miller RE, editors: Zoo and wild animal medicine, ed 7, St.
2006, Blackwell Publishing Professional, pp 192–198. Louis, 2012, Elsevier Saunders, pp 422–430.
4. McMannamon R, Lowenstine L: Cardiovascular disease in great 21. Vann Bonn W, Jensen ED, Brook F: Radiology, computed
apes. In Fowler ME, Miller RE, editors: Zoo and wild animal tomography and magnetic resonance imaging. In Dierauf LA,
medicine, ed 7, St. Louis, 2012, Elsevier Science, pp 408–415. Gulland FMD, editors: CRC handbook of marine mammal
5. Mikota SK, Sargent EL, Ranglack GS: The musculoskeletal medicine, ed 2, Boca Raton, FL, 2000, CRC Press, pp 557–591.
system. In Medical Management of the elephant, West Bloomfield, 22. Regnault S, Dixon JJI, Warren-Smith C, et al: Skeletal pathology
MI, 1994, Indira Publishing House, pp 137–143. and variable anatomy of elephant feet assessed using computed
6. Mumby C, Bouts T, Sambrook L, et al: Validation of a new tomography, PeerJ 5:e2877, 2017, doi:10.7717/peerj.2877.
radiographic protocol for Asian elephant feet and description of 23. Rush EM, Brawner WR, Ogburn AL, et al: Comparison of
their radiographic anatomy, Vet Rec 173:318, 2013. radiographs versus computed tomography for evaluation of the
7. Oosterhuis JE: Elephant toenail abscess workshop. In: Caring for distal limb in an Asian elephant. Proceedings of the American
Elephants Series. AAZV Annual Conference August 2004. Association of Zoo Veterinarians, 2005, pp. 68–69.
8. Sigeal-Willott JL, Alexander A, Isaza R: Digital radiography 24. Rush EM, Shores A, Meintel S, et al: Growing skull fracture
of the elephant foot. In Fowler ME, Miller RE, editors: Zoo in a red-tailed hawk (Buteo jamaicensis), J Zoo Wildl Med
and wild animal medicine, ed 7, St. Louis, MO, 2012, Elsevier 45(3):658–663, 2014.
Science, pp 422–430. 25. Wakman KA, Sanchez CR, Lung NP, et al: The use of magnetic
9. Vann Bonn W, Brook F: Overview of diagnostic imaging. In resonance imaging to better define hoof pathology in the reticu-
Dierauf LA, Gulland FMD, editors: CRC handbook of marine lated giraffe (Giraffa camelpardalis reticulata), J Zoo Wildl Med
mammal medicine, ed 2, Boca Raton, FL, 2000, CRC Press, pp 45(3):668–671, 2014.
551–556. 26. Van Zeeland YRA, Shoemaker NJ, Hsu EW: Advances in diag-
10. Wiedner E: Proboscidea. In Fowler ME, Miller RE, editors: Zoo nostic imaging. In Speer BL, editor: Current therapy in avian
and wild animal medicine, ed 8, St. Louis, 2015, Elsevier Science, medicine and surgery, 2016, St. Louis, MO, pp 531–549.
pp 517–532. 27. Capello V, Cauduro A, Widmer WR: The basics of radiology.
11. Adkesson MJ, Rabin DA: Degenerative skeletal disease of pri- In Capello V, Lennox AM, editors: Clinical radiology of exotic
mates. In Fowler ME, Miller RE, editors: Zoo and wild animal companion mammals, Ames, IA, 2008, Wiley-Blackwell, pp
medicine, ed 7, St. Louis, 2012, Elsevier Science, pp 396–407. 2–43.
12. Silverman S: Diagnostic imaging. In Mader DR, editor: Reptile 28. Morgan JP: Exotic species radiology. In Techniques of veterinary
medicine and surgery, ed 2, St. Louis, MO, 2006, Saunders, radiology, Ames, IA, 1994, Iowa State University Press, pp
Elsevier, pp 471–489. 441–453.
13. Brook F, Vann Bonn W, Jensen E: Ultrasonography. In Dierauf 29. Wisner E, Zwingenberger A: Atlas of small animal CT and MRI,
LA, Gulland FMD, editors: CRC handbook of marine mammal Ames, IA, 2015, Wiley-Blackwell.
medicine, ed 2, Boca Raton, FL, 2000, CRC Press, pp 593–620. 30. Morgan JP: Avian radiology. In Techniques of veterinary radiology,
14. Cooley SD, Schlipf JW, Jr, Stieger-Vanegas SM: Computed Ames, IA, 1994, Iowa State University Press, pp 433–440.
tomographic characterization of the pulmonary system in clini- 31. Wilkinson R, Hernandez-Divers S, LaFortune M, et al: Diag-
cally normal alpacas, Am J Vet Res 74:572–578, 2013. nostic imaging techniques. In McArthur S, Wilkinson R, Meyer
15. De Rycke LM, Gielen IM, Simoens PJ, et al: Computed tomog- J, editors: Medicine and surgery of tortoises and turtles, Ames, IA,
raphy and cross-sectional anatomy of the thorax in clinically 2004, Blackwell, pp 187–238.
normal dogs, Am J Vet Res 66:512–524, 2005. 32. Silverman S: Diagnostic imaging. In Mader DR, editor: Reptile
16. Henninger W: Use of computed tomography in the diseased medicine and surgery, ed 2, St. Louis, MO, 2006, Saunders,
feline thorax, J Small Anim Pract 44:56–64, 2003. Elsevier, pp 471–489.
17. Kealy JK, McAllister H, Graham JP: The radiograph. In Diag- 33. Malka S, Hawkins MG, Jones JH, et al: Effect on body position
nostic radiology and ultrasonography of the dog and cat, ed 5, St. on respiratory system volume in anesthetized red-tailed hawks
Louis, 2011, Elsevier, Saunders, pp 1–22. (Buteo jamaicensis) as measured via computed tomography, Am J
18. Morandi F, Mattoon JS, Lakritz J, et al: Correlation of helical Vet Res 70(9):1155–1160, 2009.
and incremental high-resolution thin-section computed tomo- 34. Stickle RL, Hathcock JT: Interpretation of computed tomo-
graphic imaging with histomorphometric quantitative evaluation graphic images, Vet Clin North Am Small Anim Pract 23:417–435,
of lungs in dogs, Am J Vet Res 64:935–944, 2003. 1993.
19. Prather AB, Berry CR, Thrall DE: Use of radiography in
combination with computed tomography for the assessment
SECTION 8
226
33
Equine Herpesviruses and
Interspecies Infections
ALEX DAVID GREENWOOD AND KLAUS OSTERRIEDER
Equine Herpesvirus Background and neurologic disorders in domestic equids, albeit with dif-
ferent frequencies and severities. EHV-1 is one of the most
The Herpesvirales, as a viral group, have a remarkably wide consequential pathogens of domestic horses (Equus caballus)
host species range and can infect countless vertebrate worldwide.6 In horses there are serious consequences of
and invertebrate species.1 While not among the most EHV-1 infection as show events, races, and transport
aggressive and virulent viruses known, within each of the may be affected, with the economic consequences often
herpesvirus families, subfamilies, and genera are pathogens, devastating due to movement restrictions, and cancelling
which may cause a range of clinical signs and have serious of events.7
consequences for the host. Despite the wide distribution Considered a horse pathogen, recent discoveries in both
of herpesviruses among the metazoa, comfort has been zoo and wildlife strongly suggest that EHV-1 and its close
taken in the widely accepted tenet that herpesviruses are relative EHV-9 lack strong species specificity and can both
generally host-specific and, therefore, zoonotic infections infect and cause disease in a wide variety of mammalian
are not a general consideration for this viral group.2 taxa (Fig. 33.1).8–13
However, this tenet is eroding as examples of historical
and recent cross-species (and even cross-ordinal) infections Diagnostics
have been described, many associated with severe symp-
toms or fatalities; for example, malignant catarrhal fever Because of the lack of species specificity of EHV-1 and
(MCF) is caused by a member of the Gammaherpesvirinae EHV-9 and clinical consequences, the rest of this chapter
that is not restricted to a specific species and can cause will focus on these two virus species. Before discussing the
outbreaks with high mortality in mixed-species zoological specifics of interspecies EHV-1 infections, it is worthwhile
collections.3 reviewing the typical clinical signs and the diagnostic tools
Equine herpesviruses (EHVs) were recently shown to also available for confirming infection. Neurologic manifestation
cause interspecies infections. There are nine EHVs identi- such as epileptiform seizures are an indicator of encephalitis,
fied to date, three of which (EHV-2, EHV-5, EHV-7) are which in captive settings could be taken as evidence for
classified within the Gammaherpesvirinae and six of which EHV-1 or EHV-9 involvement.7 However, the list of known
(EHV-1, EHV-3, EHV-4, and EHV-6, EHV-8, EHV-9) encephalitis-causing bacterial and viral pathogens is quite
are classified within the Alphaherpesvirinae.4 EHVs classified long. In polar bears (Ursus maritimus), domestic cats, and
as Gammaherpesvirinae are rarely associated with disease. humans, autoimmune encephalitis against glutamate recep-
However, the gammaherpesviruses in particular appear to tors has been documented.14 Therefore, while encephalitis
have historically violated the tenet of expected host specific- is a commonly observed symptom in EHV-1 and EHV-9
ity, with most viruses in this group deriving from bats interspecies infections, it is not a basis from which to make
and primates.5 The lack of direct or frequent association a diagnosis. In addition, the neurologic disease induced
with negative health outcomes (MCF, Epstein-Barr virus, by EHV-1 in horses is an encephalopathy, a stroke-like
and Kaposi sarcoma-associated herpesviruses as exceptions syndrome following endothelial cell infection and vasculitis,
with few equine associated health effects) is reflected in not a bona fide encephalitis.
less research focused on equine gammaherpesviruses and There are some considerations in suspected EHV-1 and
the long- or short-term health consequences of interspecies EHV-9 cases for facilitating diagnostics and helping to
infection are unclear. ensure that a conclusive diagnosis can be reached. In non-
The Alphaherpesvirinae EHV-1, EHV-4, and EHV-9 are fatal cases where abortion occurs or neurologic symptoms
all associated with abortion, neonatal death, and respiratory are detected, fetal tissues should be frozen for diagnostic
227
228 SE C T I O N 8 Emerging and Changing Infectious Diseases
• Figure 33.1 Equine herpesviruses (EHV-1 and EHV-9) infect a broad range of mammal species
experimentally, in captivity, and in nature. Infected host species are indicated by picture or by vector
graphic. Reservoir species for EHV-1 and EHV-9 are boxed in grey. The asterisk “*” indicates that while
EHV-1 and EHV-9 circulate among wild and domestic equids, their distributions are not uniform. For
example, the domestic horse breeds are not reservoirs for EHV-9. Experimentally infected animal groups
are boxed in blue and nonexperimental infections (zoological collections or in the wild) are boxed in orange.
EHV-1 and -9 have been detected in both wild and captive zebra and rhinoceros species. However, the
only known fatal EHV-associated rhinoceros case was an Indian rhinoceros (Rhinoceros unicornis). The
black bear (Ursus americanus), Thompson’s gazelle (Eudorcas thomsonii), zebra, domestic cat, domestic
dog, house mouse, and hamster photographs were kindly provided by Arne Lawrenz, Oliver Höner, Marion
East and Peter Seeber, Kristin Mühldorfer, Emanuel Heitlinger and Satoko Izume, respectively. The black
bear, macaque, and rabbit photographs were kindly provided by the Tierpark Berlin and the goat, llama,
giraffe, pig, rhinoceros, and water buffalo photos by the Zoological Garden Berlin.
CHAPTER 33 Equine Herpesviruses and Interspecies Infections 229
purposes. From adults, nasal discharge, nasal swabs, blood, when bound to the primary antibody, initiates an enzymatic
and serum should be collected. In fatal cases, lung, blood, reaction that can be detected by determining substrate
serum, nasal epithelia, and brain tissue should be removed conversion using an optical scanner that measures optical
and stored at −80°C if possible and not at −20°C. Of critical density in the wells of the plate. If no antibody is present
importance, samples should be collected and stored, and in the serum, there is nothing for the secondary antibody to
preferably frozen, as quickly as possible to prevent sample bind to and no signal is emitted. In the case of EHV-1 and
degradation, which complicates or prevents further down- EHV-9, discrimination was achieved, although peptides
stream analysis. In many cases it is better to just refrigerate for EHV-1 cross-react somewhat with EHV-9-specific sera.
samples if they can be transported to a diagnostic laboratory However, the developed EHV-9 peptide does not react
within a few days. with EHV-1 positive sera.17 The peptides were shown to
In equids, pathology and histology usually present as the be specific and applicable to blood samples from a variety
myeloencephalopathy, mostly affecting the spinal cord. In of wild and captive animals. They were also substantially
non-equids, neurologic symptoms are often characterized more sensitive to low titer antibody than SNT analysis. Of
by severe seizures. Pathologic findings generally include note, due to issues of background and secondary antibody
nonsuppurative encephalitis and gliosis, but Barr bodies are cross reaction, application to carnivore sera has not been
mostly absent.8,10 DNA extracted from tissues can be tested successful to date, restricting analysis in these animals to
for presence of herpesviruses using pan-herpesvirus (qPCR) SNT. Nonetheless, specific peptide-based ELISA systems
protocols,15 which can distinguish between herpesviruses provide a very efficient method for screening serum from
normally associated with a given species or evidence for a animals for past exposure to EHVs. Both methods allow for
cross-species transmission following DNA sequencing of the determination of EHV-1 and EHV-9 exposure, whether
amplified PCR products. More sensitive and EHV-specific the individual is currently infected or not.
quantitative PCR assays16 can be applied in cases of faint
signal or to quantitate viral loads in a given sample with Experimental Interspecies Infections
such services provided by many laboratories in Europe and
the United States. In the case of EHV-1, there are a number Evidence for the general ability of EHV-1 and EHV-9 to
of World Organisation for Animal Health (OIE) reference cause disease comes in part from experimental infection
laboratories that provide state-of-the-art diagnostic services studies (Fig. 33.1). Recent work has shown that much
(http://www.oie.int/). like gammaherpesviruses, EHV-1 and EHV-9 can infect
Symptomatic individuals in a population, and even non-equine species. Several experimental infections have
viremic individuals, are only a fraction of animals that have demonstrated that a wide range of species in most mamma-
been infected at some point in their lifetimes. Even for lian orders are susceptible to EHV-1 and EHV-9. Relatively
highly pathogenic viruses, many infected individuals will few experimental infections in non-equid species have been
be asymptomatic and will clear the infection. However, performed for EHV-1. Rabbits experimentally infected with
infection generally results in the production of specific EHV-1 developed both respiratory and neurologic symp-
antibodies that may persist in the individual long term or, in toms.19 Laboratory mice infected with EHV-1 could clear
some cases, lifelong. Therefore, exposure of different species lytic infection but latency was established. However, this
to different EHVs can be measured indirectly, irrespective of depended on the EHV-1 strain with some strains (Brazilian
whether the individual is currently productively infected or strains A4/72 and A9/92) able to induce neurologic clinical
not. Serology-based approaches measure antibodies against signs.20,21
a given virus or viral antigen. Assays based on whole viruses, EHV-9 experimental infection studies are broader with
for example, serum neutralization tests (SNTs), examine respect to representatives of different mammalian orders
whether viral replication can be neutralized by serum from tested. Laboratory mice were able to clear experimental
a given animal, indicating antibodies against the virus are infection after 72 hours.22 However, lesions were induced in
produced by the individual. However, SNTs will fail to suckling hamsters.23 Neurologic signs were observed in both
discriminate closely related viruses, for example, EHV-1 dogs and cats infected with EHV-9.24,25 Lethal encephalomy-
from EHV-4 or EHV-9, because they are genetically and elitis was induced by EHV-9 in goats.26 Conflicting results
antigenically very similar. Recently, synthetic peptides that were observed for pigs. In one study, no clinical signs were
are designed to specifically identify viral epitopes that are observed, although neurologic lesions were observed, while
as distinct as possible have been developed and tested for in a second experimental infection meningoencephalitis was
their discriminatory power for the closely related alpha- induced.27,28 It is not clear why the outcomes were variable.
herpesviruses EHV-1, EHV-4, and EHV-9.17,18 The test Among primates, macaques (Macaca macaca) appeared to be
used is an antigen enzyme-linked immunosorbent assay resistant to EHV-9 infections, whereas marmosets exhibited
(ELISA) and can be performed on 96 samples in a single severe neurologic symptoms.29,30 The conclusions that can
experiment in microwell plastic plates, allowing for the be drawn are that EHV-1 and particularly EHV-9 are able
simultaneous screening of a large number of individual to infect and cause clinical symptoms in a broad diversity
animals. The peptide antigens are used to coat the plates and of mammalian orders, including primates, in experimental
serum is applied followed by a secondary antibody, which, infections.
230 SE C T I O N 8 Emerging and Changing Infectious Diseases
Nonexperimental Interspecies Infections that captive zebras have a significantly lower prevalence
of seropositivity than wild zebras. Zebras in general are
Experimental infections involved controlled and often very more likely to be serologically positive for EHV-1 when
high viral doses that may not reflect the conditions required compared to EHV-9; in contrast, rhinoceros are more likely
for natural viral transmission. In addition, progression of to be serologically positive for EHV-9 than EHV-1. The
infection is often very specific to the tissues infected at results suggest that African rhinoceros may be reservoirs
the point of entry for the virus, for example, intranasal species for EHV-1 and particularly EHV-9. This may have
administration of viruses. Therefore, while many species can significant management repercussions as even in zoological
potentially be infected by EHV-1 and EHV-9, it remains collections that do not maintain zebras or other equids
somewhat unclear what the full species susceptibility spec- in captivity, the presence of rhinoceros could potentially
trum of infected may be in natural settings or in zoological result in cross-species transmission of EHV-1 or EHV-9.
collections. We make the distinction between natural set- Although diagnosed fatal cases are not frequent, in captivity
tings and zoological collections because natural infections and in the wild, EHV-1 and -9 exhibit a broad host range
would involve contact among members of the same species and broad prevalence in perissodactyls.
or among species that are sympatric in nature. In contrast,
zoological collections bring together many species at high Modes of Transmission
densities, many of which have never existed in sympatry
and are potentially mutually naïve to pathogens carried by EHV-1 and EHV-9 infection is known to be spread from
species in adjacent enclosures. It should also be pointed individual to individual as a respiratory smear infection.
out that it is far easier to observe clinical signs resulting However, this route of transmission applies to equids and
from interspecific infections in zoological collections in may not be generally applicable to interspecies transmis-
general, because there is little predation that would result sions. In the majority of observed cross-species transfers,
in the culling of sick animals. Individual animals are also there was no obvious physical contact between equids and
under a level of surveillance that is rarely achieved in the non-equids. For example, in a 2010 fatal EHV-1 infection
wild, facilitating the observation of even minor alteration of a polar bear, the zebra enclosure was 200 m from that of
in behavior indicative of disease. the polar bear, and the zookeepers were not shared between
The lack of host specificity of EHV-1 and EHV-9 is the enclosures.10 Nor were the polar bears fed equine meat
not restricted to experimental infection and extends to or carcasses, which could expose them to remaining infec-
both zoological collections and the wild. Among equids, tious virus if the culled equids were virus positive at the time
fatal cases of cross-species transmissions in zoos have been of killing. The conditions were similar in a fatal polar bear
observed frequently in a diverse set of taxa. Among non- case in 2009 with involvement of EHV-9.12 Transmission
equid species, EHV-9 was isolated from fatal cases in captive was indirect inasmuch as neither equids nor rhinoceros were
Thompson’s gazelles (Eudorcas thomsonii), giraffes, and a co-housed in close proximity. In a recent case, a polar bear
polar bear.9,11,12 Although identified in non-equid species, and Indian rhinoceros exhibited neurologic signs. The polar
EHV-9 does not appear to be a naturally occurring virus bear recovered and was found weakly positive for EHV-1
of domestic horses though it can be isolated from several in nasal swabs.8 Several weeks later, the rhino aborted an
wild equine species, particularly zebras.31–33 Captive gazelles, 8.5-month-old fetus, exhibited neurologic clinical signs,
antelopes, cattle, alpacas (Vicugna pacos), llamas (Lama and was euthanized after failure to respond to treatment.
glama), and black bears (Ursus americanus) have also been An EHV-1–EHV-9 recombinant very closely related to that
shown to be infected with EHV-1.13,34–36 More recently, an found in the 2010 polar bear case was identified in the
EHV-1–EHV-9 recombinant virus was associated with both lung and brain of the rhinoceros. A clear route of spread
lethal and nonlethal polar bear encephalitis cases and with or connection, if there was any, between the polar bear
a fatal encephalitis case in an Indian rhinoceros (Rhinoceros enclosure and rhino enclosure is not obvious. Clearly, direct
unicornis).8,10 respiratory transmission from equid to non-equid has not
In the wild, EHV-1 and EHV-9 also exhibit cross-species been likely in the majority of reported cases.
circulation where they have been tested (which to date Fomites, defined as nonliving objects (anything from
has been relatively limited). Serologic evidence exists that dust to clothing to shared instruments) are often responsible
black and white rhinoceros (Diceros bicornis and Cerato- for transfer of pathogens.12 Supporting this mode of trans-
therium simum, respectively) are seropositive for EHV-1 mission, Saklou et al. (2013) demonstrated that in “barn
and EHV-9.37 A recent study of 277 animals in the wild conditions” and at certain temperatures, infectious EHV-1
or kept in captivity, including 151 wild samples from could persist on a variety of surfaces including stall bedding,
41 wild plains zebras (Equus quagga) and 17 wild black shavings, and leather.38
rhinoceros sera, indicated significant prevalence of EHV-1 Another potential source of contamination is water.
and EHV-9 in both captive and wild populations.17 The EHV-1 remains stable for several weeks in water, even at
results demonstrated very high prevalence of EHV-1 and high temperature.39,40 Many zoological collections have
EHV-9 in zebras and black and white rhinoceros in both enclosure-connecting water sources or noncaptive animal
captivity and the wild. Statistical analysis demonstrated populations (rodents, birds) that move among enclosures
CHAPTER 33 Equine Herpesviruses and Interspecies Infections 231
and/or water sources. Thus, it is possible that water con- management and hygiene options available. Diagnostics for
taminated with the viruses could expose multiple enclosures viremic and nonviremic are rapid, sensitive, and specific.
to various EHVs. Alternatively, secondary reservoirs or If used for regular monitoring, shedding can be detected
fomite carrying rodents and birds could transfer infected early enough to isolate individual animals and prevent the
materials from one water source to the next. build-up of virus in the environment that would promote
The mode of transmission from environmental sources interspecies transfer. Implementation of such monitoring is
or intermediate hosts remains an unsolved mystery and strongly recommended.
there may be more than one route. This is a critical area
of research because, without a clearer picture, developing
management strategies will remain difficult. Regardless, References
stringent hygiene controls should be applied to prevent
EHVs from spreading from equid (or rhinoceros) enclosures 1. Fields BN, editor: Virology. In Virology, Philadelphia, 2007,
to other areas of the zoo. Equid meat should be fed only Lippincott Williams & Wilkins.
to carnivores naturally sympatric with equids. For example, 2. Tischer BK, Osterrieder N: Herpesviruses–a zoonotic threat? Vet
lions hunt and consume zebras in nature but polar bears do Microbiol 140:266–270, 2010.
not. Therefore, it is likely less problematic, from a health 3. Zanin E, Capua I, Casaccia C, et al: Isolation and characteriza-
standpoint, to feed zebra or other equid meat to lions than tion of Aujeszky’s disease virus in captive brown bears from Italy,
J Wildl Dis 33:632–634, 1997.
to non-African carnivores. This applies to mixed species
4. Davison AJ, Eberle R, Ehlers B, et al: The order Herpesvirales,
enclosures where natural sympatry should be considered Arch Virol 154:171–177, 2009.
when placing species together. 5. Escalera-Zamudio M, Rojas-Anaya E, Kolokotronis SO, et al:
Bats, primates, and the evolutionary origins and diversification
To Vaccinate or Not? of mammalian gammaherpesviruses, MBio 7:2016.
6. Pusterla N, Hussey GS: Equine herpesvirus 1 myeloencephalopa-
While EHV-1 has been intensively studied in domestic thy, Vet Clin North Am Equine Pract 30:489–506, 2014.
horses, there are still sufficient unknowns about the virus, 7. Lunn DP, Davis-Poynter N, Flaminio MJ, et al: Equine
for example, its capacity to evade hosts immune responses, herpesvirus-1 consensus statement, J Vet Intern Med 23:450–461,
to preclude a highly effective vaccination strategy.7 Among 2009.
the recommendation for horses are a commercial live viral 8. Abdelgawad A, Azab W, Damiani AM, et al: Zebra-borne equine
herpesvirus type 1 (EHV-1) infection in non-African captive
vaccine (Rhinoimmune) and two inactivated viral vaccines
mammals, Vet Microbiol 169:102–106, 2014.
(Pneumabort K-Fort Dodge and Prodigy-Intervet).7 There 9. Fukushi H, Tomita T, Taniguchi A, et al: Gazelle herpesvirus 1:
is some conflicting evidence as to whether live or inactivated a new neurotropic herpesvirus immunologically related to equine
vaccines can lower incidence of EHV-1 induced abortions, herpesvirus 1, Virology 227:34–44, 1997.
but the majority of publications suggest efficacy. There is 10. Greenwood AD, Tsangaras K, Ho SY, et al: A potentially fatal
very little evidence, however, that vaccination can prevent mix of herpes in zoos, Curr Biol 22:1727–1731, 2012.
neurologic disease. It is clear, however, that vaccination does 11. Kasem S, Yamada S, Kiupel M, et al: Equine herpesvirus type
not guarantee the complete prevention of viral shedding. 9 in giraffe with encephalitis, Emerg Infect Dis 14:1948–1949,
Vaccination trials will have to be performed in different 2008.
equid species to determine the efficacy of the commercial 12. Schrenzel MD, Tucker TA, Donovan TA, et al: New hosts for
vaccines and determine if the benefits of a vaccination equine herpesvirus 9, Emerg Infect Dis 14:1616–1619, 2008.
13. Wohlsein P, Lehmbecker A, Spitzbarth I, et al: Fatal epizootic
program are worth the financial cost. Regardless of whether
equine herpesvirus 1 infections in new and unnatural hosts, Vet
vaccination regimens are implemented or not, consistent Microbiol 149:456–460, 2011.
monitoring of viral shedding, isolation of shedding indi- 14. Pruss H, Leubner J, Wenke NK, et al: Anti-NMDA receptor
viduals, and decontamination of areas exposed to shedding encephalitis in the polar bear (Ursus maritimus) Knut, Sci Rep
animals will be key to managing cross-species transmission. 5:12805, 2015.
If live virus escape can be reduced at the source, the chance 15. VanDevanter DR, Warrener P, Bennett L, et al: Detection and
of interspecies transmission should also be reduced. analysis of diverse herpesviral species by consensus primer PCR,
J Clin Microbiol 34:1666–1671, 1996.
16. Perkins GA, Goodman LB, Dubovi EJ, et al: Detection of equine
Conclusions herpesvirus-1 in nasal swabs of horses by quantitative real-time
EHV-1 and EHV-9 have the ability to infect a very wide PCR, J Vet Intern Med 22:1234–1238, 2008.
17. Abdelgawad A, Hermes R, Damiani A, et al: Comprehensive
range of mammals in experimental, captive, and natural
serology based on a peptide ELISA to assess the prevalence of
settings. The full “natural” host range of these viruses is closely related equine herpesviruses in zoo and wild animals,
not fully determined and the modes of transmission need PLoS ONE 10:e0138370, 2015.
further exploration. It seems evident that the two viruses 18. Lang A, de Vries M, Feineis S, et al: Development of a peptide
can threaten the health of a variety of equid and non-equid ELISA for discrimination between serological responses to
mammals in zoological collections. Vaccination efficacy in equine herpesvirus type 1 and 4, J Virol Methods 193:667–673,
nondomestic equid species needs research, but there are other 2013.
232 SE C T I O N 8 Emerging and Changing Infectious Diseases
19. Kanitz FA, Cargnelutti JF, Anziliero D, et al: Respiratory and 9 in common marmosets (Callithrix jacchus), J Med Primatol
neurological disease in rabbits experimentally infected with equid 36:335–342, 2007.
herpesvirus 1, Microb Pathog 87:45–50, 2015. 31. Borchers K, Bottner D, Lieckfeldt D, et al: Characterization of
20. Baxi MK, Borchers K, Bartels T, et al: Molecular studies of the equid herpesvirus 1 (EHV-1) related viruses from captive Grevy’s
acute infection, latency and reactivation of equine herpesvirus-1 zebra and blackbuck, J Vet Med Sci 68:757–760, 2006.
(EHV-1) in the mouse model, Virus Res 40:33–45, 1996. 32. Ghanem YM, Fukushi H, Ibrahim ES, et al: Molecular phy-
21. Mori CMC, Mori E, Favaro LL, et al: Equid herpesvirus type-1 logeny of equine herpesvirus 1 isolates from onager, zebra and
exhibits neurotropism and neurovirulence in a mouse model, J Thomson’s gazelle, Arch Virol 153:2297–2302, 2008.
Comp Pathol 146:202–210, 2012. 33. Ibrahim ES, Kinoh M, Matsumura T, et al: Genetic relatedness
22. El-Nahass E, El-Dakhly KM, El-Habashi N, et al: Susceptibility and pathogenicity of equine herpesvirus 1 isolated from onager,
of BALB/c-nu/nu mice and BALB/c mice to equine herpesvirus zebra and gazelle, Arch Virol 152:245–255, 2007.
9 infection, Vet Pathol 51:581–590, 2014. 34. Chowdhury SI, Ludwig H, Buhk HJ: Molecular biological
23. El-Habashi N, El-Nahass el S, Fukushi H, et al: Experimental characterization of equine herpesvirus type-1 (Ehv-1) Isolates
intranasal infection of equine herpesvirus 9 (EHV-9) in suckling from ruminant hosts, Virus Res 11:127–139, 1988.
hamsters: kinetics of viral transmission and inflammation in the 35. Crandell RA, Ichimura H, Kit S: Isolation and compara-
nasal cavity and brain, J Neurovirol 16:242–248, 2010. tive restriction endonuclease DNA fingerprinting of equine
24. Yanai T, Fujishima N, Fukushi H, et al: Experimental infection herpesvirus-1 from cattle, Am J Vet Res 49:1807–1813, 1988.
of equine herpesvirus 9 in dogs, Vet Pathol 40:263–267, 2003. 36. Rebhun WC, Jenkins DH, Riis RC, et al: An epizootic of blind-
25. Yanai T, Tujioka S, Sakai H, et al: Experimental infection ness and encephalitis associated with a herpesvirus indistinguish-
with equine herpesvirus 9 (EHV-9) in cats, J Comp Pathol able from equine herpesvirus-i in a herd of alpacas and llamas, J
128:113–118, 2003. Am Vet Med Assoc 192:953–956, 1988.
26. Taniguchi A, Fukushi H, Yanai T, et al: Equine herpesvirus 9 37. Fischer-Tenhagen C, Hamblin C, Quandt S, et al: Serosurvey for
induced lethal encephalomyelitis in experimentally infected selected infectious disease agents in free-ranging black and white
goats, Arch Virol 145:2619–2627, 2000. rhinoceros in Africa, J Wildl Dis 36:316–323, 2000.
27. Narita M, Uchimura A, Kawanabe M, et al: Invasion and spread 38. Saklou N, Burgess B, Morley PW, et al: Equine herpesvirus type
of equine herpesvirus 9 in the olfactory pathway of pigs after 1 (EHV-1) in the environment: how long will it stay infective?
intranasal inoculation, J Comp Pathol 124:265–272, 2001. Equine Vet J 45:16, 2013.
28. Narita M, Uchimura A, Kimura K, et al: Brain lesions and 39. Doll ER, Mc CW, Bryans JT, et al: Effect of physical and chemi-
transmission of experimental equine herpesvirus type 9 in pigs, cal environment on the viability of equine rhinopneumonitis
Vet Pathol 37:476–479, 2000. virus propagated in hamsters, Cornell Vet 49:75–81, 1959.
29. Kodama A, Yanai T, Kubo M, et al: Cynomolgus monkeys 40. Dayaram A, Franz M, Schattschneider A, et al: Long term stabil-
(Macaca fascicularis) may not become infected with equine ity and infectivity of herpesviruses in water, Sci Rep 21(7):46559,
herpesvirus 9, J Med Primatol 40:18–20, 2011. 2017. doi:10.1038/srep46559
30. Kodama A, Yanai T, Yomemaru K, et al: Acute neuropatho-
genicity with experimental infection of equine herpesvirus
34
Ebola Virus Disease in Great Apes
KENNETH CAMERON AND PATRIC IA REED
Disclaimer: Any opinions or conclusions in this article are it has been significant. Only ZEBOV and TAFV have been
those of the authors and do not necessarily represent the identified in great apes.3,5,8,9
views of the US Fish and Wildlife Service.
Ebola Virus Disease in African Apes
Introduction
In contrast to human EVD outbreaks, which have occurred
All species of great apes are classified as either endangered or throughout tropical Africa, those involving apes have been
critically endangered by the International Union for Con- restricted to West and Central Africa. In 1994 a single EVD
servation of Nature (IUCN); worldwide, great ape popula- outbreak occurred in chimpanzees in Taï National Park,
tions are in decline, threatened by habitat degradation and Ivory Coast. Mortality was estimated at approximately 25%
destruction, poaching, climate change, and disease.1 Among of the chimpanzee community.10 The pathogen involved
disease threats, Ebola virus disease (EVD) is cited as a major was identified as a new species of filovirus, subsequently
cause of population declines of western lowland gorillas named Taï Forest Ebolavirus.11 One person, who contracted
(Gorilla gorilla gorilla).2 EVD after performing a postmortem examination on a
chimpanzee that died during the outbreak, survived
Ebola Virus Disease infection.11
In Central Africa, Gabon and Congo are home to
EVD is an acute, severe disease caused by viruses of the genus the vast majority of western lowland gorillas (Gorilla
Ebolavirus, family Filoviridae.3 Zaire Ebolavirus (ZEBOV), gorilla gorilla) and central chimpanzees (Pan troglodytes
the species most associated with EVD in apes, was first troglodytes).2 In the late 1990s and early 2000s, EVD
discovered in 1976. Since then, four additional species of outbreaks involving ZEBOV occurred in two clusters in
Ebolavirus have been identified: Sudan Ebolavirus (SUDV), northeastern Gabon and northwestern Congo. Between
Taï Forest Ebolavirus (TAFV), and Bundibugyo Ebolavirus 1994 and 1996, multiple reports of gorilla, chimpanzee,
(BDBV) in Africa and Reston Ebolavirus (RESTV) in Asia.3 and other wildlife carcasses accompanied the human EVD
Multiple Ebola virus species have caused more than 30 outbreaks in Gabon’s Minkébé Forest region. Most of
EVD outbreaks in humans throughout sub-Saharan Africa. the human outbreaks there were linked to contact with
Eight of those—all caused by ZEBOV—have occurred in chimpanzee carcasses.4,12 Wildlife abundance surveys
Gabon and the Republic of Congo (Congo) and were conducted in the region before and after the outbreaks
linked to human contact with gorilla or chimpanzee meat showed up to a 98% decline in gorilla and chimpanzee
and/or carcasses.4,5 A prolonged EVD outbreak caused by populations.12
ZEBOV in West Africa from 2013 to 2016 infected more Five human EVD outbreaks that occurred in the
than 28,000 people, and more than 11,000 people died.6 Gabon-Congo transborder region from 2001 to 2005 were
One epidemiologic study proposed that the index case of also associated with contact with infected carcasses, mostly
that epidemic may have been exposed to ZEBOV from gorillas and chimpanzees.5,8,13 A large number of wildlife
an insectivorous bat; however, no definitive link could be carcasses had been reported in this region just prior to
demonstrated.7 the human outbreaks and, in each, the human index case
(usually a hunter) was linked to infection from a gorilla,
chimpanzee, or duiker carcass.5,8 Large mammal abun-
Impact of Ebola Virus Disease dance estimates conducted in 2000 and 2003 (pre- and
on Great Apes post-EVD outbreaks) in the Gabon-Congo transborder
region showed a 59%, 89%, and 53% decline in gorillas,
Although the precise degree of impact of EVD on great ape chimpanzees, and duikers, respectively. Based on this infor-
populations is difficult to quantify, there is little doubt that mation, investigators estimated that hundreds or thousands
233
234 SE C T I O N 8 Emerging and Changing Infectious Diseases
of gorillas there were lost to EVD.8 Between 2001 and Clinical Signs
2003, investigators recovered 98 wildlife carcasses in this
region, including 65 great apes. Of 21 carcasses tested for In humans, the incubation period of EVD is typically 2–21
Ebola virus by polymerase chain reaction (PCR), antigen days, and victims are not considered infectious until they
detection, and, in some cases, immunohistochemical develop symptoms. Once symptoms develop, they are vari-
staining, 14 tested positive by at least one diagnostic able but include a sudden onset of fever, fatigue, muscle
test, including 10 gorillas, 3 chimpanzees, and 1 duiker.5 pain, headache, and sore throat, followed by vomiting,
From 2002 to 2003, researchers in the Lossi Gorilla diarrhea, rash, and, in a minority of cases, both internal
Sanctuary in northwestern Congo witnessed a mortal- and external bleeding (e.g., oozing from the gums, blood
ity event in free-ranging wild gorillas and chimpanzees in the stool).22 Sequelae in many EVD survivors—including
under study. Over a period of several months, dozens of musculoskeletal pain, headache, ocular symptoms, abdomi-
gorilla and chimpanzee carcasses were discovered. Overall nal pain, and sexual dysfunction—constitute what is referred
mortality, based on nest counts and observations of to as post-Ebola syndrome. These symptoms may persist for
known individuals, was estimated at 90%–95%. Gorilla months to years after recovery.23
mortality there was estimated to have exceeded 5000 In apes, clinical signs are not specifically known, but
individuals.14 there have been observations of wild chimpanzees exhibiting
Then in 2005 an EVD outbreak occurred in great apes severe diarrhea and emaciation, and other nonhuman pri-
in Odzala-Kokoua National Park (OKNP), northwestern mates exhibiting vomiting, diarrhea, hair loss, emaciation,
Congo, concurrent with a nearby human outbreak. Wildlife and epistaxis in association with known ZEBOV-associated
abundance surveys conducted in the park in 2005, 2008, EVD epizootics.24 Necropsy results on a TAFV-positive
and 2012 estimated a nearly 50% reduction in gorillas there, chimpanzee reported signs of hemorrhage and nonclotting
representing about 20,000 individuals.15 This decrease in blood.11
gorilla populations in the absence of observed outbreaks In one study, nonape primates experimentally infected
supports the assertion that, given the extreme difficulty with ZEBOV or SUDV exhibited a wide variety of clinical
in monitoring wild great ape populations in this region, signs, typically beginning within 3 days postinfection. These
additional outbreaks may have gone undetected. In fact, included fever, anorexia, cachexia, dehydration, diarrhea,
an unusual ape mortality event was noted in early 2007 intermittent melena, rectal bleeding, and petechial rash.25 In
along the eastern edge of OKNP, during which local villag- another study, nonape primates died within 8 days of being
ers reported dozens of gorilla carcasses. One chimpanzee experimentally infected with ZEBOV and within 12–14
and nine gorilla carcasses were sampled over a 4-month days after being infected with TAFV.26
period.16 Despite the lack of confirmatory testing, the Apes may survive Ebola virus infection, as evidenced by
mortality event appeared to be consistent with an EVD the detection of antibodies in wild-born chimpanzees and
outbreak (K. Cameron, P. Reed, personal communication, other primates, in feces of wild gorillas, and by the survival
June 30, 2017). of individuals in groups that suffered mortality during an
Although multiple human EVD outbreaks have occurred EVD outbreak.14,24,27,28
in the Democratic Republic of Congo (DRC), none has
been reported in bonobos (Pan paniscus) or eastern gorillas Transmission
(Gorilla beringei). To date, BDBV and SUDV have not been
detected in wild apes. In humans, Ebola viruses are highly infectious. Infection
Based on observations of EVD impacts on known wild usually occurs through direct contact of infected bodily
gorilla populations, ZEBOV-associated mortality in apes fluids with mucous membranes or broken skin. An increased
may reach 95%, exceeding that in humans.14,17 In summary, risk of transmission occurs in the acute phase of infection,
although the precise number of apes killed by EVD is when patients are viremic. Prior to the onset of clinical
impossible to determine, overwhelming circumstantial signs, and once the virus is cleared, there is little risk of
evidence points to massive EVD-associated gorilla and transmission.
chimpanzee mortality in the Central Africa region. However, ZEBOV may persist in urine as well as in the
placenta, amniotic fluid, and fetus in women who became
Ebola Virus Disease in Asian Apes infected while pregnant; it may also persist in breast milk
in women infected while breastfeeding.22 Viral persistence
RESTV, the only Ebola virus species known to occur in Asia, has also been demonstrated in “privileged” sites, such as the
has been identified in laboratory nonape primates imported eyes, brain, and testes in both convalescing patients and
to the United States from the Phillipines.18 RESTV is not laboratory monkeys.29,30 ZEBOV has been isolated from
believed to be pathogenic to humans or Asian monkeys and human semen up to 7–9 months postonset of EVD.31 Such
no cases of EVD have been reported in Asian apes.19 One viral persistence raises the possibility of transmission or
study did report ZEBOV antibodies in captive Bornean reemergence postoutbreak.31
orangutans (Pongo pygmaeus), but these findings have been Routes of transmission in apes have not been definitively
called into question.20,21 established, including routes of spillover from putative
CHAPTER 34 Ebola Virus Disease in Great Apes 235
reservoir species. A predominant theory proposes that closely related to Ebolavirus—has been isolated from Egyp-
apes may consume food items contaminated with bodily tian fruit bats (Rousettus aegyptiacus), supporting the theory
fluids (saliva, urine, feces) from reservoirs (e.g., fruit bats) of bats as filovirus reservoirs.52,53
at common feeding sites.32,33 However, viral shedding of
ZEBOV in wild bat urine or feces has not been reported.34 Diagnostics
Transmission within ape groups presumably occurs
through physical contact with a sick or deceased group The diagnosis of Ebola virus disease in humans is based on
member.5 Transmission between ape groups may occur the presence of viral RNA, antigen, or antibodies. Diag-
to varying degrees by a variety of mechanisms, including nostic assays employed in suspected human cases include
contact with a deceased member of another group, disper- antibody-capture enzyme-linked immunosorbent assay
sion of females upon the death of the group leadership, (ELISA), antigen-capture detection tests, serum neutraliza-
cofeeding encounters between groups, intergroup aggressive tion test, reverse transcriptase polymerase chain reaction
interactions, or intergroup breeding.5,35–37 (RT-PCR) assay, electron microscopy, and virus isolation
Evidence suggests that EVD outbreaks in ape popula- by cell culture.22 Preferred diagnostic specimens in humans
tions may extend over large geographic areas, though how include whole blood in ethylenediaminetetraacetic acid
this occurs is a matter of debate. Two theories predominate (EDTA) from live symptomatic patients or an oral fluid
in the literature.5,12,14 One proposes that transmission of the specimen in universal transport medium collected from a
virus continues from group to group, moving as a “wave” deceased patient or when blood collection is not possible.22
of transmission across the landscape.38 Transmission in this Collection of blood samples from wild apes is impractical
scenario would be largely dependent on the incubation and diagnostics are usually conducted on carcass samples.
period and speed of disease progression in individual apes, Obtaining an accurate diagnosis may be particularly chal-
on group and individual movement, and on intra- and lenging in tropical field settings, where carcasses degrade
intergroup social dynamics.39,40 But detection of different quickly. Degraded tissues may be unsuitable for many
strains of ZEBOV in gorilla carcasses located in close Ebola virus diagnostic assays and more prone to produce
proximity during a given outbreak argues against a major false-negative results.5 In fact, in field settings, attempts at
role of such transmission.5 viral isolation from decomposed carcasses have generally
A second theory proposes that such wide geographic been unrewarding.
spread results from multiple spillover events into an ape
population from a reservoir host species. Transmission in Differential Diagnosis
this scenario would be more dependent on reservoir species
dynamics.8 The fact that large distances between ZEBOV- Differential diagnosis is dependent on the epidemiologic
positive ape carcasses found during a short period and the context and may include bacterial, viral, fungal, and parasitic
existence of physical barriers to ape movement, such as causes.32 In humans, significant diseases to rule out include
roads and rivers, tend to support this mode of transmis- malaria, typhoid fever, shigellosis, cholera, leptospirosis,
sion.5 These proposed transmission mechanisms are not plague, rickettsiosis, relapsing fever, meningitis, hepatitis,
mutually exclusive, however, and could occur concurrently yellow fever, and other viral hemorrhagic fevers.22 In African
during a given outbreak. apes, anthrax may also be considered.54,55
development specifically for use in great apes, is based on protective equipment that burial of a ZEBOV-infected
an ape-specific cytomegalovirus (CMV).58 This is a self- carcass—particularly in a wooded environment—would
replicating vaccine, designed to spread from ape to ape, put investigators at additional risk of exposure. Incineration
stimulating immunity against ZEBOV as is does so. The using vegetation and/or diesel fuel, which has been employed
proposed advantages of such a vaccine are (1) that it could, in more accessible regions, is very time-consuming, imprac-
once administered to relatively few individuals, immunizing tical in many remote regions, and carries additional risk.
a much larger portion of the ape population and (2) that Transporting the carcass to another location for incineration
it could do so with reduced effort and in a logistically or burial is highly ill-advised, as it carries significant risk
easier and more cost-effective, manner. The predominant and goes against basic principles of pathogen containment.
potential disadvantage of this self-replicating vaccine Spraying the carcass with a 5% sodium hypochlorite solu-
relates to concerns about safety in target and nontarget tion has also been proposed but carries the risk of human
species.59 poisoning in the event that local inhabitants attempt to
Practical concerns about vaccinating wild apes include scavenge meat (K. Cameron, P. Reed, personal communica-
the route of vaccine administration (e.g., by injection versus tion, June 30, 2017).
orally), the ease of access to apes (e.g., habituated to close Given these concerns, one program left carcasses to decay
human presence versus unhabituated), the number of doses unburied following sampling (K. Cameron, P. Reed, personal
of the vaccine needed to achieve adequate immunity in an communication, June 30, 2017). Concerns about human
individual (i.e., a single dose versus the need for booster infection in the meantime were addressed by conducting
doses), the purpose of the vaccination campaign (e.g., outreach education in local communities before departing
to provide immunity from future infection or to halt an the region (K. Cameron, P. Reed, personal communication,
ongoing epizootic), the relative risk of an EVD outbreak to June 30, 2017).
a given ape population, and more.59 A given vaccine may
have more or less application to various ape populations, Future Directions
their level of habituation, and the degree and nature of the
Ebola virus threat. Given that many human EVD outbreaks Although laboratory and field studies over the past 4
are closely associated with contact with wildlife carcasses, decades have greatly improved our understanding of the
reducing occurrence of EVD in ape populations could also epidemiology and pathophysiology of Ebola virus in
have protective benefits for humans. humans, our understanding of its ecology as well as its
This issue of ape vaccination against Ebola virus has precise impact on wild apes remains relatively poor. This
stimulated lively discussion among conservationists. Moral lack of basic knowledge makes designing potential mitiga-
issues about intervening in natural processes in “wild” tion strategies for both apes and humans more challenging.
populations versus human obligations to do so, potential More investigation is needed to better characterize the basic
unintended consequences of the use of genetically modified ecology of Ebola viruses, importantly regarding potential
vaccines, and other vaccine safety issues dominate these reservoir species, including bats; the identification of poten-
discussions.59,60 First and foremost, any vaccine must be tial vector species that may complicate the transmission
shown to be efficacious and safe for target (ape) and non- chain and obfuscate the reservoir host; elucidation of the
target (nonape, including wildlife and human) species prior ecology of those reservoirs and vectors; and identification
to deployment.60 of environmental factors that may affect the virus’s sporadic
reemergence. Greater understanding of the virus’s transmis-
Carcass Disposal sion in a natural setting is needed, including the route or
routes of spillover from reservoir to susceptible hosts; both
In the event of detection of a great ape carcass that may interspecific and intergroup transmission among apes; and
signal an EVD epizootic, programs have been established the potential roles of other susceptible species, such as wild
to collect samples for Ebola virus diagnostic testing.5,16 pigs and duikers. As this knowledge improves, there will
Whether or not to dispose of these potentially Ebola virus– be a need for updated and more refined projections of the
infected carcasses in order to limit transmission potential long-term impacts of EVD on ape populations. Improved
has been debated and remains a practical issue in EVD surveillance, allowing early detection of EVD outbreaks in
surveillance efforts. Carcass degradation is likely to vary apes, would help to facilitate much of the needed research.
with environmental conditions. One study showed that In the meantime, work should continue on the potential
carcasses are likely to remain infective for 3–4 days in a use of vaccination as a control measure to protect ape
tropical environment.8 In another study, in which carcasses species against EVD, with a particular focus on vaccine
were not exposed to insect scavengers, viable Ebola virus was safety for both target and nontarget species. In order to
isolated from infected laboratory macaques up to 7 days or accomplish this and maximize future successes in preserving
more postmortem.61 these iconic ape species, the issue of EVD’s impact on wild
Burial and/or incineration of carcasses on site has been ape populations will require improved open, rational, and
proposed (K. Cameron, P. Reed, personal communica- constructive dialogue within the scientific and conservation
tion, June 30, 2017). The high risk of breach of personal communities.
CHAPTER 34 Ebola Virus Disease in Great Apes 237
40. Ryan SJ, Jones JH, Dobson AP: Interactions between social 52. Towner JS, Amman BR, Sealy TK, et al: Isolation of genetically
structure, demography, and transmission determine disease diverse Marburg viruses from Egyptian fruit bats, PLoS Pathog
persistence in primates, PLoS ONE 8:e76863, 2013. 5(7):e1000536, 2009.
41. Pourrut X, Kumulungui B, Wittmann T, et al: The natural 53. Amman BR, Carroll SA, Reed ZD, et al: Seasonal pulses of
history of Ebola virus in Africa, Microbes Infect 7(7):1005–1014, Marburg virus circulation in juvenile Rousettus aegyptiacus bats
2005. coincide with periods of increased risk of human infection, PLoS
42. Swanepoel R, Leman PA, Burt FJ, et al: Experimental inocula- Pathog 8(10):e1002877, 2012.
tion of plants and animals with Ebola virus, Emerg Infect Dis 54. Leendertz FH, Lankester F, Guislain P, et al: Anthrax in Western
2:321–325, 1996. and Central African great apes, Am J Primatol 68(9):928–933,
43. Peterson AT, Bauer JT, Mills JN: Ecologic and geographic 2006.
distribution of filovirus disease, Emerg Infect Dis 10:40–47, 2004. 55. Hoffmann C, Zimmermann F, Biek R, et al: Persistent anthrax as
44. Peterson AT, Carroll DS, Mills JN, et al: Potential mammalian a major driver of wildlife mortality in a tropical rainforest, Nature
filovirus reservoirs, Emerg Infect Dis 10:2073–2081, 2004. 548(7665):82, 2017.
45. Peterson AT, Lash RR, Carroll DS, et al: Geographic potential 56. De Santis O, Audran R, Pothin E, et al: Safety and immuno-
for outbreaks of Marburg hemorrhagic fever, Am J Trop Med Hyg genicity of a chimpanzee adenovirus-vectored Ebola vaccine in
75:9–15, 2006. healthy adults: a randomised, double-blind, placebo-controlled,
46. Sanchez A, Geisbert TW, Feldmann H: Filoviridae: marburg dose-finding, phase 1/2a study, Lancet Infect Dis 16:311–320,
and ebola viruses. In Knipe DM, Howley PM, editors: Fields 2016.
virology, Philadelphia, 2007, Lippincott Williams & Wilkins, 57. Henao-Restrepo AM, Camacho A, Longini IM, et al: Efficacy
pp 1279–1304. and effectiveness of an rVSV-vectored vaccine in preventing
47. Hayman DTS, Yu M, Crameri G, et al: Ebola virus antibodies in Ebola virus disease: final results from the Guinea ring vaccina-
fruit bats, Ghana, West Africa, Emerg Infect Dis 18:1207–1209, tion, open-label, cluster-randomised trial (Ebola Ça Suffit!),
2012. Lancet 389(10068):505–518, 2017.
48. Leroy EM, Kumulungui B, Pourrut X, et al: Fruit bats as reser- 58. Marzi A, Murphy AA, Feldmann F, et al: Cytomegalovirus-based
voirs of Ebola virus, Nature 438(7068):575–576, 2005. vaccine expressing Ebola virus glycoprotein protects nonhuman
49. Pourrut X, Souris M, Towner JS, et al: Large serological survey primates from Ebola virus infection, Sci Rep 6:21674, 2016.
showing co-circulation of Ebola and Marburg viruses in Gabo- 59. Leendertz SA, Wich SA, Ancrenaz M, et al: Ebola in great apes –
nese bat populations, and a high seroprevalence of both viruses current knowledge, possibilities for vaccination, and implications
in Rousettus aegyptiacus, BMC Infect Dis 9(1):159, 2009. for conservation and human health, Mamm Rev 2016.
50. Ohimain EI: Ecology of ebolavirus: a review of current 60. Osofsky S, Olson S, Cameron K: [edited] PRO/AH/EDR>
knowledge, speculations and future research directions, Virology Ebola update (35): vaccine, comment. 14 Apr 2016. Available
1(3):555564, 2016. at http://promedmail.org/post/4167038.
51. Strong JE, Wong G, Jones SE, et al: Stimulation of Ebola virus 61. Prescott J, Bushmaker T, Fischer R, et al: Postmortem stability
production from persistent infection through activation of the Ras/ of Ebola virus, Emerg Infect Dis 21(5):856, 2015.
MAPK pathway, Proc Natl Acad Sci USA 105(46):17982–17987,
2008.
35
Chagas Disease: Wildlife Infection
With Trypanosoma Cruzi in a One
Health Context
SARAH HAMER AND CAROLYN HODO
239
240 SE C T I O N 8 Emerging and Changing Infectious Diseases
• Figure 35.1 Key reservoir hosts in the ecology of Chagas disease across the Americas differ in
domestic, peridomestic, and sylvatic transmission settings, with marked geographic variation in vertebrate
reservoir community and triatomine vector distribution. However, there is considerable overlap between
cycles, with dogs playing important roles in both domestic and peridomestic transmission, and certain
wildlife species being important as both sylvatic and peridomestic reservoirs. True domestic transmission
involves vectors that colonize homes, feeding on human and domestic animal inhabitants, while perido-
mestic and sylvatic transmission occur in the outside environment and involve different vector species.
domestic settings.3,13 Dogs can develop acute or chronic arthropods. Furthermore, many zoo animals have extensive
cardiac disease, whereas the clinical implications of infec- travel histories, providing opportunities for the importation
tion in other domestic reservoirs have been relatively under- or exportation of parasite infection to and from the park,
studied. In the United States, T. cruzi has been detected in including exotic parasite strains that may differ from those
at least 26 wildlife species across 15 southern states, as far maintained by local wildlife. Disease has been reported
north as Missouri.2,14 Among the most well-studied and in a number of wild and captive New and Old World
highly infected reservoirs in the United States are carnivores nonhuman primates.17–21 For example, heart abnormalities
including raccoons (Procyon lotor), striped skunks (Mephitis consistent with Chagas disease were found at a nonhuman
mephitis), coyotes (Canis latrans), gray foxes (Urocyon cine- primate center in Louisiana, where 1.6% of over 2000
reoargenteus); woodrats (Neotoma spp.); opossums (Didelphis individuals had been exposed to the parasite.22 Addition-
virginiana); and nine-banded armadillos (D. novemcinctus). ally, 8.5% of cynomolgus macaques (Macaca fascicularis) in
In Brazil, key wildlife reservoirs include opossums (Didelphis outdoor housing in central Texas were infected or exposed
spp. and Philander spp.); xenarthrans including armadillos, to the parasite, many of which harbored cardiac lesions
anteaters, and sloths; nonhuman primates especially tama- from infection.19 At a nonhuman primate facility in central
rins (Leontopithecus spp.); carnivores especially coatis (Nasua Texas with ongoing T. cruzi infections in macaques, we
nasua); and bats; while diverse rodent species generally have found high levels of parasite in the blood and tissues of
overall low T. cruzi infection prevalence and seem to play native wildlife trapped at the facility, including raccoons,
a secondary role as reservoirs.15 In Mexico, opossums (D. opossums, and skunks, suggesting these wild animals serve
marsupialis and D. virginiana), rodents, bats, raccoons, and as a source of infection that can be transmitted by vectors
xenarthrans are among the key reservoir hosts in nature.16 to the primates.23 A polar bear (Ursus maritimus) at a zoo
Zoological parks or areas with captive animals are poten- in Mexico developed fatal acute cardiomyopathy due to T.
tially high-risk areas for Chagas disease if they coincide with cruzi infection.24 Chagas disease was implicated as the cause
the distribution of triatomine vectors, because animals are of sudden death of a 7-year-old female wolf-hybrid at a zoo
housed in outdoor areas that are accessible by blood-sucking in central Texas where triatomine vectors are commonly
CHAPTER 35 Chagas Disease: Wildlife Infection With Trypanosoma Cruzi in a One Health Context 241
found in and around the animal enclosures (Clark P, per- appearing to reflect local, easily-accessible blood sources,13
sonal communication, February 27, 2017). Because Chagas including nidicolous mammals such as woodrats and
disease often presents with nonspecific signs or sudden armadillos.2 Because infection prevalence31 and sylvatic host
death, it is likely that it is underreported in wildlife and associations2 are known to vary among triatomine species,
zoo animal species. risk of parasite transmission to humans and animals likely
varies according to the vector species found in a local area.
Vectors
Transmission Routes
Across the Americas, insects of the family Reduviidae, sub-
family Triatominae, including species of the genera Rhodnius, Transmission of T. cruzi is predominantly through the
Panstrongylus, and Triatoma, are the most important vectors infected feces of the triatomine vector. The insect vector
of T. cruzi (Fig. 35.2) to humans and animals. These noc- acquires the blood form trypomastigote stage of the parasite
turnal insects are blood feeding throughout all five nymphal while feeding on an infected mammalian host, and the para-
stages and as adults. Eleven species of triatomine insects have site replicates as the insect form epimastigote stage in the
been recorded across the southern United States,2 where digestive tract of the bug. Epimastigotes mature to infective
they are colloquially known as “kissing bugs” or “conenose metacyclic trypomastigotes in the insect hindgut, which are
bugs.” In Mexico, 19 of the 31 local triatomine species have passed in the feces and may be deposited onto a susceptible
been consistently found to invade human houses and all host. The parasite does not penetrate intact skin but can
have been found to be naturally infected with T. cruzi.25 In enter microlesions at the site of feeding, other broken skin,
the Amazon Basin, at least 10 of the 16 triatomine species or a mucous membrane; this mode of transmission is termed
that occur in the region have been found to be infected stercorarian, or “vector-fecal” and is thought to be the most
with T. cruzi.26 Both adults and nymphs have been collected important route accounting for human infections across the
from zoos,24,27,28 nonhuman primate centers,29 and domestic Americas where vectors colonize homes. Transmission via
animal enclosures.30 Blood meal analyses of triatomines the oral route occurs through a variety of means and may
have revealed a generalist opportunistic feeding strategy be the most important route in free-ranging mammals.5
Oral transmission can occur following the ingestion of an
infected bug, ingestion of food contaminated with bug feces,
or ingestion of infected blood or tissue from a parasitemic
host. Additionally, at least one species of opossum (Didelphis
marsupialis) has been demonstrated to shed the infective
form of the parasite in anal sac secretions,32 presenting a
method for oral transmission via ingestion of parasite from
opossum feces in contaminated foodstuffs. Oral transmis-
sion has been demonstrated in skunks, raccoons, opossums,
and wood rats,33–36 and suggested in dogs.37 Additionally,
congenital transplacental transmission has been described
in humans and dogs,38 but its importance as a route of
infection to wildlife or other animals in natural environ-
ments has not been studied. Congenital transmission was
not demonstrated in experimentally infected opossums (D.
marsupialis).39
It has been suggested that hematophagous (sucking) lice
may serve as alternative vectors of T. cruzi in nonhuman
primate facilities, though transmission via this route has
not been proven.40 Bed bugs (Cimex lectularius)41 and bat
bugs (Cimex pilosellus)42 have been shown to be competent
• Figure 35.2 Triatomine vectors, commonly known as kissing bugs vectors for T. cruzi in laboratory settings, but the impor-
or conenose bugs, are large nocturnal insects that are dark brown tance of other blood-sucking vectors in the transmission
to black in body color, often with distinct, reddish- to cream-colored of T. cruzi in natural settings has yet to be demonstrated.
stripes visible along the edges of the abdomen. Their head is stick-like
and tapering, and their six legs are relatively thin and tapering. Left to
right; Triatoma protracta, the most common species in the western Implications of Trypanosoma cruzi
United States; Triatoma gerstaeckeri, the most common species in Genetic Diversity
Texas; Triatoma sanguisuga, the most common species in the eastern
United States. Scale bar represents 25 mm or approximately 1 inch. T. cruzi is a genetically heterogeneous species and comprises
(Photo credit: Gabriel Hamer, PhD, Texas A&M University Department
at least seven strain types or discrete typing units (DTUs):
of Entomology. Originally printed in Curtis-Robles R, Wozniak EJ,
Auckland LD, et al: Combining public health education and disease TcI-VI and TcBat. TcI has been divided into TcIdom and
ecology research: using citizen science to assess Chagas disease TcIsyl, representing domestic and sylvatic isolates.43 The
entomological risk in Texas. PLoS Negl Trop Dis 9:e0004235, 2015.) parasite DTUs provide a framework for understanding
242 SE C T I O N 8 Emerging and Changing Infectious Diseases
associations of the parasite with different geographical prominent feature, especially right bundle branch block.
locations, reservoir host species, and clinical manifestations Dependent edema may be a feature, and scrotal edema was
in hosts infected with different parasite strain types.3,44 described in an infected chimpanzee.20 In some cases, acute
The general distribution of DTUs includes TcI across the death occurs with no premonitory clinical signs.
Americas; TcII, TcV, and TcVI in the Southern Cone; TcIII
in central South America; and TcIV in the southern United Pathology
States and northern South America.45 In some geographic
areas, parasite DTUs are closely associated with certain The typical T. cruzi pathologic changes in the heart are
vector or host species; for example, the vast majority of T. characterized by mononuclear cell inflammatory infiltrates
cruzi in raccoons in the southern United States is TcIV,46,47 composed of lymphocytes, plasma cells, and macrophages,
and opossums across the Americas harbor TcI.2,3 TcIII together with myocardial degeneration and necrosis, with
occurs almost exclusively where the vector Panstrongylus fibrosis in the chronic stages. The heart may be grossly
geniculatus is distributed in central South America. enlarged and/or pale depending on the severity and chronic-
Growing evidence suggests that certain T. cruzi strain ity of the disease. The amastigote tissue stages (“leishmanial
types may be associated with different clinical outcomes forms”) of the parasite are found in intracellular tissue pseu-
in humans.13,45,48 Similarly, experimental studies in dogs docysts, are approximately 4 µm in diameter, and contain
have demonstrated differing clinical, pathologic, and a round nucleus and rod-shaped kinetoplast. Amastigotes
immunologic outcomes resulting from infection with dif- are most commonly found in the heart (Fig. 35.3) but
ferent strains. For example, dogs infected with T. cruzi have been reported in many other tissues, such as skeletal
isolates from an armadillo and opossum developed acute muscle, lymph nodes, central nervous system tissue, and
and chronic myocarditis, while dogs infected with an isolate tissues of the digestive tract. Atypical manifestations of T.
from another dog did not develop disease,49 and increased cruzi infection may include acute lymphadenopathy51 or
numbers of inflammatory cells were observed in the heart neurologic disease as described in dogs52,53 and a horse.54
in dogs infected with TcI compared to TcII.50 Few studies While studies that document wildlife infection are
have assessed the degree to which different T. cruzi strain common, those that characterize the pathology associated
types are associated with differential clinical outcome in with infection are rarer. Experimentally infected South
infected wildlife species. American opossums (D. marsupialis) developed only mild
inflammation associated with scattered parasites, while more
Pathogenesis intense inflammation and tissue destruction was observed
in naturally infected opossums.55 Mild to moderate inflam-
Although few specific studies have examined T. cruzi patho- mation was observed in tissues of experimentally infected
genesis in wildlife species, in humans and dogs, infection Virginia opossums (D. virginanus), while experimentally
with T. cruzi is characterized by acute, indeterminate, and infected raccoons (P. lotor) developed moderate to severe
chronic stages. During the acute stage, parasitemia can be
high and the parasite is found in many organs. A small
percentage of individuals experience severe cardiac disease
during this stage, but the majority will have only mild
nonspecific clinical signs or be completely asymptomatic.
An effective immune response reduces parasite numbers
but is unable to eliminate the parasite completely. During
the indeterminate phase, clinical signs are absent, but the
parasite persists in target organs, most often the heart, and
antibodies can be detected in the serum. Approximately
30% (in humans) of indeterminate stage patients will
progress into the chronic stage characterized by cardiomy-
opathy or rarely, megasyndromes of the gastrointestinal
tract. The proportion of infected nonhuman animals that
develop chronic disease is relatively unknown and likely
species-dependent.
• Figure 35.3 Photomicrograph of the heart of a naturally infected
Clinical Signs rhesus macaque from a nonhuman primate facility in central Texas.
A cardiac myocyte contains a pseudocyst filled with Trypanosoma
Clinical signs during the acute phase, if present, are gener- Cruzi amastigotes (arrow). The myocardium is infiltrated by inflam-
matory cells, predominantly lymphocytes and plasma cells with few
ally nonspecific. In the chronic stage, clinical signs are those
macrophages, and there is multifocal myocardial degeneration and
of heart failure (exercise intolerance, lethargy, coughing, loss (hematoxylin and eosin stain, 20× magnification). (Photo credit:
pleural effusion, ascites, dependent edema), and not spe- Wallace Baze, DVM, PhD, DACVP, Keeling Center for Comparative
cific to T. cruzi infection. Cardiac arrhythmia may be a Medicine and Research, MD Anderson Cancer Center.)
CHAPTER 35 Chagas Disease: Wildlife Infection With Trypanosoma Cruzi in a One Health Context 243
inflammation.56 No histopathologic lesions were observed vary widely across tests and species, and dynamics of local
in a survey of infected raccoons, coyotes, foxes, and bobcats transmission vary by geographic location, affecting positive
in Texas,46 while a more detailed pathologic investigation predictive values. Serologic tests detect the presence of anti-
revealed inflammation and myocardial degeneration in bodies; antibody-positive animals are generally considered to
some infected Texas coyotes.57 also be currently infected because self-cure is thought to be
rare. Serologic testing, namely, indirect fluorescent antibody
Diagnosis assay (IFA), is a common tool for detecting anti-T. cruzi
antibodies in dogs but is not commonly available for zoo
Diagnosis of T. cruzi presents significant challenges, espe- and wildlife species. Rapid immunochromatographic lateral
cially for zoo and wild animal species for which diagnostic flow assays designed for human diagnostics have been used
tests have not been validated. Diagnosis of Chagas disease on dog and wildlife samples in research settings. Limitations
is based on clinical suspicion and supporting diagnostic test of serology include the difficulty of finding species-specific
results. The parasite may be directly observed via microscopic controls for many wildlife and zoo species, lack of validation
examination of blood smears during parasitemic stages or of in these species, and potential for cross-reaction with other
infected tissues during chronic stages. Parasitemia has been trypanosome species in areas where those are endemic (e.g.,
shown to occur within days to 4 weeks after infection in dogs T. rangeli, T. vivax, T. evansi, T. equiperdum, and T. theileri).
and mice but becomes undetectable after a short period of Further, very acute infections may fail to be detected using
time.58 On light microscopy, the blood form of the parasite serologic methods alone; anti-T. cruzi antibodies are first
is C- or S-shaped, 16–22 µm in length, with a rod-shaped detected an average of 3 weeks postinfection in dogs.58
kinetoplast, an undulating membrane, free flagellum, and a Polymerase chain reaction (PCR) allows direct detection
round nucleus located in the central or front portion of the of the parasite, is generally considered highly sensitive and
body (Fig. 35.4). Histopathologic examination of tissues specific, and does not necessitate species-specific reagents
can demonstrate the presence of amastigotes and/or char- or controls, thereby increasing its utility as applied to
acteristic lesions (described previously). Amastigotes may be wildlife or zoo animals. However, PCR of blood is a useful
rare in chronic stages of the disease and their absence does diagnostic tool only during a period of parasitemia, the
not preclude diagnosis if characteristic myocardial inflam- level and duration of which may vary between host species.
mation is present. Immunohistochemistry would be helpful PCR of the heart or other tissues is useful for postmortem
to confirm infection when amastigotes are not seen but is diagnosis. Often, several diagnostic methods must be used
not widely available. to make a diagnosis, combining clinical suspicion, serologic
Numerous indirect methods are used to determine infec- status, pathology, and molecular results.
tion, many of which have not been properly validated for
use in wildlife species or domestic animal species, given Treatment, Management, Prevention
the absence of a gold standard diagnostic test. Sensitivity
and specificity of different existing diagnostic tests may Treatment options for T. cruzi infection are extremely
limited. Benznidazole and nifurtimox are the antiparasitic
drugs most commonly used to treat humans but are not
readily available in veterinary medicine (especially not
in the United States) and are associated with significant
side effects. Clinical case management of diseased animals
typically focuses on the symptomatic treatment of cardiac
complications, and case management is complicated by the
absence of a vaccination or approved antiparasitic treat-
ment against T. cruzi. Although many infected animals may
remain asymptomatic, there are not currently prognostic
data available to evaluate which animals will develop disease.
Major recommendations for reducing risk of transmis-
sion in domestic and captive animal settings generally focus
on integrated vector control methods (Box 35.1). Vector
control recommendations include insecticides, screening
animal enclosures, reducing use of night lighting (which
may attract flying adult triatomines), removing brush piles
and other microhabitats conducive to wildlife nests that
serve as sources of blood meals for kissing bugs, and sealing
• Figure 35.4 Trypanosoma cruzi trypomastigotes in a canine blood
crevices that may serve as hiding places for triatomines.
smear, with undulating membrane, free flagellum, apical nucleus, and
large kinetoplast (100× magnification). (Photo credit: Karen Snowden, Additionally, because of the possibility of transmission via
DVM, PhD, DACVM, Texas A&M University College of Veterinary carnivory, efforts should be made to limit the exposure
Medicine and Biomedical Sciences.) of captive animals to potentially infected wildlife. While
244 SE C T I O N 8 Emerging and Changing Infectious Diseases
• BOX 35.1 Recommendations for Triatomine that allows them to serve as reservoirs to infect vectors, as
Vector Control has been shown for dogs and cats in villages in Argentina.62
However, this contribution must be considered in light of
• Use of residual insecticides and physical barriers such as the presence and infection status of other local reservoirs
diatomaceous earth
• Screening animal enclosures and human housing
and vectors.
• Reducing use of night lighting (which may attract flying adult
triatomines) Conclusion
• Removing brush piles and other microhabitats conducive to
wildlife nests or bug harborage sites T. cruzi is a zoonotic parasite with a wide host range
• Sealing crevices within animal enclosures that may serve as
hiding places for triatomines
including hundreds of wild and domestic mammals, and a
One Health approach to assess human, wildlife, domestic
animal, and vector infection data has been useful in defin-
ing transmission networks and assessing disease risk. The
clinical impacts of infection across most wild taxa are largely
unknown. Because most diagnostic tests are validated for
rats captured at Texas nonhuman primate facilities with human or canine use, diagnosis of Chagas disease in zoo and
documented local transmission were not infected with T. wild animals is a challenge; further, antiparasitic treatments
cruzi,59 rodents have been implicated as reservoirs in other are not widely available especially in the United States. Vigi-
areas.13,60 Because of the ability of the opossum to shed the lance for triatomine vectors and integrated vector control
infective stage of the parasite through anal secretions,32 care are the key for Chagas disease control. Enhanced awareness
should also be taken to prevent the contamination of animal for vectors and infected animals among veterinary practitio-
feed by opossum feces. ners working with zoo animals and wildlife will afford the
While direct animal-animal transmission is considered protection of both human and animal health.
unlikely in the absence of predation, there is a potential
risk that captive infected animals may serve as reservoirs to
infect local vectors. However, this level of risk is not well References
characterized, and an individual animal’s reservoir potential
is variable on a species-level and individual-animal basis. 1. Gaunt M, Miles M: The ecotopes and evolution of triatomine
bugs (Triatominae) and their associated trypanosomes, Mem Inst
Additionally, in endemic areas including the southern
Oswaldo Cruz 95:557–565, 2000.
United States, local wildlife populations are frequently 2. Bern C, Kjos S, Yabsley MJ, et al: Trypanosoma cruzi and Chagas’
highly infected, likely serving as the primary parasite disease in the United States, Clin Microbiol Rev 24:655–681,
reservoirs, with captive animal reservoirs playing secondary 2011.
roles. Nevertheless, vector control activities could help to 3. Jansen AM, Xavier SCC, Roque ALR: The multiple and complex
mitigate risk by limiting the access of vectors to infected and changeable scenarios of the Trypanosoma cruzi transmission
captive animals. cycle in the sylvatic environment, Acta Trop 151:1–15, 2015.
4. Chagas C: Nova tripanozomiaze humana: estudos sobre a mor-
folojia e o ciclo evolutivo do Schizotrypanum cruzi n. gen., n. sp.,
Zoonotic Potential ajente etiolojico de nova entidade morbida do homem, Mem Inst
The main risk of human exposure to T. cruzi is from Oswaldo Cruz 1:159–218, 1909.
5. Jansen AM, Roque ALR, Xavier SCC: Trypanosoma cruzi
contact with infected vectors. The efficiency of the stercorar-
enzootic cycle: general aspects, domestic and synanthropic hosts
ian (vector-fecal) route of transmission is extremely low, and reservoirs. In Telleria J, Tibayrenc M, editors: American
and estimates are that several hundred or more contacts trypanosomiasis Chagas disease: One hundred years of research, ed
with infected bugs are necessary for vector-borne parasite 2, Cambridge, MA, 2017, Elsevier, pp 265–282.
transmission,61 and so human transmission is greatest in 6. Chagas C: Sobre un trypanosomo do tatú, Tatusia novemcincta,
areas where there are frequent contacts between vectors transmitido pela Triatoma geniculata Latr. (1811). Possibilidade
and people. Direct transmission from infected animals to do ser o tatu um depositario do tatú Trypanosoma cruzi no
humans is possible via exposure to infected blood, which mundo exterior Brasil, Médico 1:305–306, 1912.
may occur through broken skin (needle stick, existing skin 7. World Health Organization: Chagas disease in Latin America: an
wounds) or through contact with mucous membranes (oral, epidemiological update based on 2010 estimates, Wkly Epidemiol
conjunctival). As such, care should be taken when handling Rec 6:33–44, 2015.
8. Schmunis GA, Yadon ZE: Chagas disease: A Latin American
the blood of infected animals. Additionally, because of the
health problem becoming a world health problem, Acta Trop
ability of infected opossums to shed infective parasites in 115:14–21, 2010.
their anal secretions, care should be taken when handling 9. Manne-Goehler J, Umeh CA, Montgomery SP, et al: Estimating
feces and cleaning animal enclosures to avoid contact with the burden of Chagas disease in the United States, PLoS Negl Trop
broken skin or mucous membranes. The presence of infected Dis 10:e0005033, 2016.
animals may increase local risk to other uninfected animals 10. Cantey PT, Stramer SL, Townsend RL, et al: The United States
or humans if the infected animals maintain a parasitemia Trypanosoma cruzi infection study: evidence for vector-borne
CHAPTER 35 Chagas Disease: Wildlife Infection With Trypanosoma Cruzi in a One Health Context 245
transmission of the parasite that causes Chagas disease among 29. Kasa TJ, Lathrop GD, Dupuy HJ, et al: An endemic focus of
United States blood donors, Transfusion 52:1922–1930, 2012. Trypanosoma cruzi infection in a subhuman primate research
11. Garcia MN, Aguilar D, Gorchakov R, et al: Evidence of autoch- colony, J Am Vet Med Assoc 171:850–854, 1977.
thonous Chagas disease in southeastern Texas, Am J Trop Med 30. Curtis-Robles R, Snowden KF, Dominguez B, et al: Epidemiology
Hyg 92:325–330, 2015. and molecular typing of Trypanosoma cruzi in naturally-infected
12. Desquesnes M: Veterinary aspects. In Telleria J, Tibayrenc M, hound dogs and associated Triatomine vectors in Texas, USA,
editors: American trypanosomiasis Chagas disease: One hundred PLoS Negl Trop Dis 11:e0005298–18, 2017.
years of research, ed 2, Cambridge, MA, 2017, Elsevier, pp 31. Curtis-Robles R, Wozniak EJ, Auckland LD, et al: Combin-
283–298. ing public health education and disease ecology research: using
13. Gürtler RE, Cardinal MV: Reservoir host competence and the citizen science to assess Chagas disease entomological risk in
role of domestic and commensal hosts in the transmission of Texas, PLoS Negl Trop Dis 9:e0004235, 2015.
Trypanosoma cruzi, Acta Trop 151:32–50, 2015. 32. Urdaneta-Morales S, Nironi I: Trypanosoma cruzi in the anal
14. Hodo CL, Hamer SA: Toward an ecological framework for glands of urban opossums. I-Isolation and experimental infec-
assessing reservoirs of vector-borne pathogens: wildlife reservoirs tions, Mem Inst Oswaldo Cruz 91:399–403, 1996.
of Trypanosoma cruzi across the southern United States, ILAR J 33. Davis DS, Russell LH, Adams LG, et al: An experimental infec-
2017. tion of Trypanosoma cruzi in striped skunks (Mephitis mephitis),
15. Jansen AM, Xavier SCC, Roque ALR: Ecological aspects of J Wildl Dis 16:403–406, 1980.
Trypanosoma cruzi: wild hosts and reservoirs. In Telleria J, 34. Yaeger RG: Transmission of Trypanosoma cruzi infection to opos-
Tibayrenc M, editors: American trypanosomiasis Chagas disease: sums via the oral route, J Parasitol 57:1375–1376, 1971.
One hundred years of research, ed 2, Cambridge, MA, 2017, 35. Roellig DM, Ellis AE, Yabsley MJ: Oral transmission of Trypano-
Elsevier, pp 243–264. soma cruzi with opposing evidence for the theory of carnivory, J
16. López-Cancino SA, Tun-Ku E, De la Cruz-Felix HK, et al: Parasitol 95:360–364, 2009.
Landscape ecology of Trypanosoma cruzi in the southern Yucatan 36. Ryckman RE, Folkes DL, Olsen LE, et al: Epizootiology of
Peninsula, Acta Trop 151:58–72, 2015. Trypanosoma cruzi in southwestern North America, J Med Ent
17. Monteiro RV, Baldez J, Dietz J, et al: Clinical, biochemical, and 2:87–108, 1965.
electrocardiographic aspects of Trypanosoma cruzi infection in 37. Montenegro VM, Jimenez M, Dias JCP, et al: Chagas disease in
free-ranging golden lion tamarins (Leontopithecus rosalia), J Med dogs from endemic areas of Costa Rica, Mem Inst Oswaldo Cruz
Primatol 35:48–55, 2006. 97:491–494, 2002.
18. Dickerson MF, Astorga NG, Astorga NR, et al: Chagas disease 38. Rodríguez-Morales O, Ballinas-Verdugo MA, Alejandre-Aguilar
in 2 geriatric rhesus macaques (Macaca mulatta) housed in the R, et al: Trypanosoma cruzi connatal transmission in dogs with
Pacific Northwest, Comp Med 64:323–328, 2014. Chagas disease: experimental case report, Vector Borne Zoonotic
19. Pisharath H, Zao C-L, Kreeger J, et al: Immunopathologic Dis 11:1365–1370, 2011.
characterization of naturally acquired Trypanosoma cruzi infec- 39. Jansen AM, Madeira FB, Deane MP: Trypanosoma cruzi infection
tion and cardiac sequalae in cynomolgus macaques (Macaca in the opossum Didelphis marsupialis: absence of neonatal trans-
fascicularis), J Am Assoc Lab Anim Sci 52:545–552, 2013. mission and protection by maternal antibodies in experimental
20. Bommineni YR, Dick EJ, Estep JS, et al: Fatal acute Chagas infections, Mem Inst Oswaldo Cruz 89:41–45, 1994.
disease in a chimpanzee, J Med Primatol 38:247–251, 2009. 40. Argañaraz ER, Hubbard GB, Ramos LA, et al: Blood-sucking
21. Gleiser CA, Yaeger RG, Ghidoni JJ: Trypanosoma cruzi infection lice may disseminate Trypanosoma cruzi infection in baboons,
in a colony-born baboon, J Am Vet Med Assoc 189:1225–1226, Rev Inst Med Trop Sao Paulo 43:271–276, 2001.
1986. 41. Salazar R, Castillo-Neyra R, Tustin AW, et al: Bed bugs (Cimex
22. Dorn PL, Daigle ME, Combe CL, et al: Low prevalence of lectularius) as vectors of Trypanosoma cruzi, Am J Trop Med Hyg
Chagas parasite infection in a nonhuman primate colony in 92:331–335, 2015.
Louisiana, J Am Assoc Lab Anim Sci 51:443–447, 2012. 42. Wood SF: Development of California Trypanosoma cruzi in the
23. Hodo CL, Wilkerson GK, Birkner EC, et al: Trypanosoma cruzi bat bedbug, J Parasit 37:330–331, 1951.
transmission among captive nonhuman primates, wildlife, and 43. Ramírez JD, Tapia-Calle G, Guhl F: Genetic structure of
vectors, EcoHealth 2018. In press. Trypanosoma cruzi in Colombia revealed by a high-throughput
24. Jaime-Andrade J, Avila-Figueroa D, Lozano-Kasten FJ, et al: nuclear multilocus sequence typing (nMLST) approach, BMC
Acute Chagas’ cardiopathy in a polar bear (Ursus maritimus) in Genet 14:96, 2013.
Guadalajara, Mexico, Rev Soc Bras Med Trop 30:337–340, 1997. 44. Ramirez JD, Guhl F, Umezawa ES, et al: Evaluation of adult
25. Ramsey JM, Townsend Peterson A, Carmona-Castro O, et al: chronic Chagas’ heart disease diagnosis by molecular and sero-
Atlas of Mexican Triatominae (Reduviidae: Hemiptera) and logical methods, J Clin Microbiol 47:3945–3951, 2009.
vector transmission of Chagas disease, Mem Inst Oswaldo Cruz 45. Zingales B, Miles MA, Campbell DA, et al: The revised Trypano-
110:339–352, 2015. soma cruzi subspecific nomenclature: rationale, epidemiological
26. Coura JR, Junqueira ÂCV, Fernandes O, et al: Emerging relevance and research applications, Infect Genet Evol 12:240–253,
Chagas disease in Amazonian Brazil, Trends Parasitol 18:171–176, 2012.
2002. 46. Curtis-Robles R, Lewis BC, Hamer SA: High Trypanosoma
27. Minuzzi-Souza TTC, Nitz N, Knox MB, et al: Vector-borne cruzi infection prevalence associated with rare cardiac pathology
transmission of Trypanosoma cruzi among captive neotropical among wild carnivores in central Texas, Int J Parasitol Parasites
primates in a Brazilian zoo, Parasit Vectors 9:1–6, 2016. Wildl 5:117–123, 2016.
28. Klotz SA, Schmidt JO, Dorn PL, et al: Free-roaming kissing 47. Roellig DM, Savage MY, Fujita AW, et al: Genetic variation and
bugs, vectors of Chagas disease, feed often on humans in the exchange in Trypanosoma cruzi isolates from the United States,
Southwest, Am J Med 127:421–426, 2014. PLoS ONE 8:e56198, 2013.
246 SE C T I O N 8 Emerging and Changing Infectious Diseases
48. Ramírez JD, Guhl F, Rendón LM, et al: Chagas cardiomyopathy 56. Roellig DM, Ellis AE, Yabsley MJ: Genetically different isolates of
manifestations and Trypanosoma cruzi genotypes circulating in Trypanosoma cruzi elicit different infection dynamics in raccoons
chronic chagasic patients, PLoS Negl Trop Dis 4:e899–9, 2010. (Procyon lotor) and Virginia opossums (Didelphis virginiana), Int
49. Barr SC, Gossett KA, Klei TR: Clinical, clinicopathologic, and J Parasitol 39:1603–1610, 2009.
parasitologic observations of trypanosomiasis in dogs infected 57. Hodo CL: The ecology of Trypanosoma cruzi and its mammalian
with North American Trypanosoma cruzi isolates, Am J Vet Res hosts in Texas, USA. Doctoral dissertation, Texas A&M Univer-
52:954–960, 1991. sity, 2017.
50. Duz ALC, Vieira PM, Roatt BM, et al: The TcI and TcII Try- 58. Snowden KF, Kjos SA: American trypanosomiasis. In Greene CE,
panosoma cruzi experimental infections induce distinct immune editor: Infectious diseases of the dog and cat, ed 4, St. Louis, 2012,
responses and cardiac fibrosis in dogs, Mem Inst Oswaldo Cruz Elsevier, pp 722–730.
109:1005–1013, 2014. 59. Hodo CL, Bertolini NR, Bernal JC, et al: Lack of Trypanosoma
51. Nabity MB, Barnhart K, Logan KS, et al: An atypical case of cruzi infection in urban roof rats (Rattus rattus) at a Texas facility
Trypanosoma cruzi infection in a young English Mastiff, Vet housing naturally infected nonhuman primates, J Am Assoc Lab
Parasitol 140:356–361, 2006. Anim Sci 56:1–6, 2017.
52. Berger SL, Palmer RH, Hodges CC, et al: Neurologic mani- 60. Ramsey JM, Gutiérrez-Cabrera AE, Salgado-Ramírez L, et al:
festations of trypanosomiasis in a dog, J Am Vet Med Assoc Ecological connectivity of Trypanosoma cruzi reservoirs and
198:132–134, 1991. Triatoma pallidipennis hosts in an anthropogenic landscape with
53. Kjos SA, Snowden KF, Craig TM, et al: Distribution and endemic Chagas disease, PLoS ONE 7:e46013, 2012.
characterization of canine Chagas disease in Texas, Vet Parasitol 61. Nouvellet P, Dumonteil E, Gourbière SB: The improbable
152:249–256, 2008. transmission of Trypanosoma cruzi to human: the missing link
54. Bryan LK, Hamer SA, Shaw S, et al: Chagas disease in a Texan in the dynamics and control of Chagas disease, PLoS Negl Trop
horse with neurologic deficits, Vet Parasitol 216:13–17, 2016. Dis 7:e2505–e2512, 2013.
55. Araujo Carreira JC, Jansen AM, Deane MP, et al: Histo- 62. Gürtler RE, Cécere MC, Lauricella MA, et al: Domestic dogs
pathological study of experimental and natural infections by and cats as sources of Trypanosoma cruzi infection in rural
Trypanosoma cruzi in Didelphis marsupialis, Mem Inst Oswaldo northwestern Argentina, Parasitology 134:69–82, 2007.
Cruz 91:609–618, 1996.
36
The Effects of Climate Change on
Disease Spread in Wildlife
ERIK HOFMEISTER AND CAROLINE VAN HEMERT
247
248 SE C T I O N 8 Emerging and Changing Infectious Diseases
Physical changes in the environment, such as higher scapularis has been expanding northward from the United
ambient or soil temperature, can release thermal constraints States into Canada, affecting southern and western Ontario,
that affect the life cycle of parasites, thereby allowing for Manitoba,21 and Quebec.22 Distribution of Lyme disease is
invasion and expansion of a parasite’s geographic range. predicted to continue its northward expansion to match the
One such example is the recently documented expansion shifting ranges of Ix. scapularis and the white-footed mouse
of parasitic nematodes in the Canadian Arctic.15 The lung (Peromyscus leucopus), a highly efficient reservoir host that
nematode Umingmakstrongylus pallikuukensis infects musk is constrained by winter climatic conditions.23 A similar
oxen (Ovibos moschatus) in the Arctic and has recently been northward expansion of Ix. ricinus, the vector of Borreliosis,
detected beyond its endemic range on Victoria Island. The tick-borne encephalitis, and other diseases in Europe, has
invasion and expansion of U. pallikuukensis have likely been reported in Norway24 and Sweden.25 Migrating birds
been facilitated by more permissive climatic conditions carrying feeding ticks are a likely source of tick range expan-
and increased animal movements.15 Higher summer tem- sion, with higher summertime temperatures facilitating tick
peratures allow the larvae of U. pallikuukensis to develop establishment in more northerly regions.24,26
to the infectious stage more rapidly within its intermediate The spread of other wildlife diseases and disease vectors
host, the marsh slug (Deroceras laeve), thus reducing the to higher elevations has also been linked to climate change.5
parasite’s life cycle from 2 years to 1 year.16 Increased rate of Currently, avian malaria, caused by Plasmodium relictum, is
larval development results in a higher density of infectious limited to elevations below 1500 m on the island of Hawai’i.
larvae and a longer period of time that muskoxen are at This is due to a reduction of mosquito activity at higher
risk of exposure.15,16 Historically, movements of muskoxen (cooler) elevations27 and also a longer development time for
populations between the Canadian mainland and Victoria the parasite within mosquitoes.28 Even though the forests
Island occurred sporadically but have recently become more of the island of Kaua’i are below 1500 m, native Hawaiian
common in association with icing events.15 Thus, changing avian communities have been maintained because the island
environmental conditions affect not only the life cycle of is cooler than comparable elevations on other Hawaiian
the parasite but also movement patterns of hosts, leading islands.29 However, recent surveys demonstrate an increase
to increased distribution of this lungworm. Similarly, Var- in avian malaria in native birds on Kaua’i, the presence of
estrongylus sp., a lung nematode that infects both muskoxen mosquitoes at higher elevations, and a precipitous decline
and caribou (Rangifer tarandus), has also been described on in native forest birds on the island.29 Native Hawaiian birds
Victoria Island, and available data suggest recent invasion are already threatened by introduced diseases,30 and climate-
and expansion.15 induced elevational shifts in avian malaria transmission
Climate change may also affect the development and could result in further extinctions (Fig. 36.1).31,32
associated geographic spread of disease vectors. Epizootic It is also important to note that, although less commonly
hemorrhagic disease (EHD) virus is vectored by a biting reported, climatic changes can lead to range contractions for
midge (Culicoides sonorensis) and causes disease in North select pathogens or parasites. Such a case has been predicted
America among populations of white-tailed deer (Odocoileus in North America for the Lone Star tick (Amblyomma
virginianus) and other wild cervids.17 As with other vector-
borne infectious diseases, higher summer temperatures
often result in increased vector abundance, higher viral load,
and a faster rate of viral replication in infected vectors.18
Lower summer rainfall may also lead to increased vector
abundance due to the greater exposure of mud flats, the
preferred breeding sites for midges. A record number of
EHD cases in wild ungulates were reported in 2012 from
27 US states, including states that had not previously
reported the disease.19 In 2012, the states reporting record
outbreaks of EHD all recorded above average or record
annual temperatures and, with few exceptions, below to
much below average precipitation. In addition, midges
capable of transmitting EHD were recorded for the first
time in southern Ontario,20 indicating a potential range
expansion of the vector.
Environmental changes have also recently influenced
the distribution of tick-borne diseases, for which wildlife
species often serve as reservoirs or play an important role • Figure 36.1 ‘Apapane (Himatione sanguinea), a native Hawaiian
been linked to climate warming.45–47 Toxicity resulting between marine and terrestrial species and creating new
from exposure to HABs that are caused by dinoflagellates, opportunities for disease transmission.51,52 Additionally, loss
cyanobacteria, or, in some cases, diatoms48 affect a variety of of sea ice allows for greater mixing of pathogen communities
vertebrate and nonvertebrate marine species. Specific drivers as eastern and western marine species come into contact,
behind HAB events are often not well understood but are such as is projected for the Canadian Arctic.10 Loss of sea ice
thought to be related to nutrient levels, ocean currents, may also result in seasonal changes in animal movements,
upwellings, and ocean temperature.49 The largest recorded with potential impacts on disease transmission. The first
outbreak of domoic acid, produced by the diatom Pseudo- known occurrence of avian cholera in Alaska occurred in
nitzschia australis, occurred in 2015 along the west coast 2013 and affected several marine bird species that had not
of North America and resulted in extensive strandings of previously been documented with the disease.13 Although
marine mammals and closures of clam and crab fisheries.45 the cause of the outbreak is unknown, it occurred coincident
This event was apparently initiated by anomalously warm with unusually warm water temperatures and reduced sea
ocean conditions, and subsequent laboratory and field experi- ice cover in the Bering Sea, conditions that allowed birds
ments demonstrated maximum growth rates of P. australis to remain in open water around the island for a longer
with increased temperature and higher toxin production duration (K. Kuletz, personal communication, December
with nutrient enrichment.45 In Scandinavian countries bor- 23, 2014).
dering the North Sea, regional climate warming has led to Direct and indirect effects of climate change may also
increased fresh water input to marine environments, causing alter transmission patterns of disease in wildlife through
stratification of the marine waters by salinity. Stratification, changes in population density or population structure.
along with increased nutrients contained in the run-off, has Unlike infectious diseases that result in low pathogenicity
been associated with changes in the diatom: dinoflagellate and a more chronic course, infectious agents that cause
ratio and bloom formation.46 Current predictions indicate epizootics are often potentiated by increases in the density
that the influences of warmer ocean temperatures, ocean or proportion of susceptible animals in a population. In
acidification, vertical stratification, extreme weather pat- North America, drought, one possible extreme outcome of
terns, and continued anthropogenic influences will lead to climate change, has been associated with transmission of St.
an increase in severe HAB events.8,45,47,50 Louis encephalitis to birds.53 Using models to predict past
In some instances, prevalence or severity of disease may and future cases of West Nile virus (WNV) in humans,
also be reduced as a result of climate change, especially for drought was identified as a more reliable predictor of
cases in which warmer temperatures hinder pathogen or human cases than temperature or precipitation alone.54
parasite development. Such an example has been described Drought is believed to promote congregation of birds at
in the Arctic for Ostertagia greuhneri, a common abomasal available water sources and increase the rate of host and
nematode of caribou and muskoxen.3,39 Contrary to expec- vector contact. Wild birds are variably susceptible to WNV,
tations, experimentally warmed tundra plots resulted in and population declines have been reported in several North
reduced development rates of O. greuhneri, a pattern that American species.55
was verified in the laboratory.39 Although cold is typically a In the marine environment, warmer water temperatures
more limiting factor than heat in tundra environments, O. may be contributing to sea star wasting disease (SSWD),
greuhneri appears to be highly adapted to Arctic conditions an emerging aquatic disease thought to be caused by a
and warming temperatures will likely decrease the exposure densovirus (Fig. 36.3).56 The virus has been detected in
of Arctic ungulates to this parasite.12 museum specimens dating back nearly 70 years, but a major
outbreak of SSWD occurred in 2013–2014 and resulted in
Emerging Diseases widespread mortality among a number of sea star species
from northern California to Alaska. The disease appears
Here we define emergence of novel pathogens to include to be more prevalent and more severe in warmer water,57
newly detected pathogens as well as occurrence of known and ocean temperature anomalies were strong during the
pathogens in previously unaffected taxa. The precise causes outbreak,57 although this pattern was not consistent across
of emerging diseases are often not known, but climate- all affected areas.58 The role of climate change in SSWD is
driven environmental changes may be contributing factors. still being investigated, but the frequency of outbreaks may
Recent shifts in ecosystem composition and population increase with rising ocean temperatures.
dynamics have important implications for disease spread as In addition to landscape-level impacts, climate change
severe and erratic disease outbreaks may occur if formerly has the potential to affect a variety of microclimates. Such
isolated species or populations come into contact. changes may be related to snake fungal disease (SFD), a
Rapid environmental changes, leading to new interac- newly emergent disease in North America, caused by the
tions between species at key ecologic transition zones, are fungus Ophidiomyces ophiodiicola (Fig. 36.4; see Chapter
predicted to lead to an increase in novel disease outbreaks 56).59 The fungus, which infects the epidermis of an animal
among Arctic wildlife.51,52 For example, reduction in sea ice and can also invade the subcutaneous tissue, muscles, and
along the Arctic coastline has led to more frequent use of organs, has been identified since 2006 in wild snakes in the
land by polar bears and walruses, thus promoting contact eastern United States.60 Infection can lead to death of the
CHAPTER 36 The Effects of Climate Change on Disease Spread in Wildlife 251
• Figure 36.3 Diseased (left) and healthy (right) ochre sea stars (Pisaster ochraceus). A large outbreak of sea star wasting disease occurred in
2013–2014 on the northern coast of Washington and was correlated with higher ocean temperatures. (Photograph by K. Lafferty, USGS Western
Ecological Research Center.)
burgdorferi, the agent of Lyme disease, at the northeastern limit populations associated with the coral Pocillopora damicornis,
of its distribution, Evol Appl 7:750–764, 2014. Front Microbiol 6:432, 2015.
24. Hasle G, Bjune G, Edvardsen E, et al: Transport of ticks by 42. Kleypas JA, Yates KK: Coral reefs and ocean acidification,
migratory passerine birds to Norway, J Parasitol 95:1342–1351, Oceanography 22:108–117, 2009.
2009. 43. Byrne M, Ho M, Selvakumaraswamy P, et al: Temperature, but
25. Jaenson T, Jaenson D, Eisen L, et al: Changes in the geographical not pH, compromises sea urchin fertilization and early develop-
distribution and abundance of the tick Ixodes ricinus during the ment under near-future climate change scenarios, Proc Biol Sci
past 30 years in Sweden, Parasit Vectors 5:8, 2012. 276:1883–1888, 2009.
26. Ogden NH, Lindsay LR, Hanincova K, et al: Role of migratory 44. Edmunds PJ, Carpenter RC: Recovery of Diadema antillarum
birds in introduction and range expansion of Ixodes scapularis reduces macroalgal cover and increases abundance of juvenile
ticks and of Borrelia burgdorferi and Anaplasma phagocytophilum corals on a Caribbean reef, Proc Nat Acad Sci U S A 98:5067–5071,
in Canada, Appl Environ Microbiol 74:1780–1790, 2008. 2001.
27. Ahumada JA, Lapointe D, Samuel MD: Modeling the population 45. Paerl HW, Huisman J: Climate change: a catalyst for global
dynamics of Culex quinquefasciatus (Diptera: Culicidae), along an expansion of harmful cyanobacterial blooms, Environ Microbiol
elevational gradient in Hawaii, J Med Entomol 41:1157–1170, Rep 1:27–37, 2009.
2004. 46. Edwards M, Johns DG, Leterme SC, et al: Regional climate
28. LaPointe DA, Goff ML, Atkinson CT: Thermal constraints to change and harmful algal blooms in the northeast Atlantic,
the sporogonic development and altitudinal distribution of avian Limnol Oceanogr 51:820–829, 2006.
malaria Plasmodium relictum in Hawai’i, J Parasitol 96:318–324, 47. Glibert PM, Icarus Allen J, Artioli Y, et al: Vulnerability of
2010. coastal ecosystems to changes in harmful algal bloom distribu-
29. Paxton EH, Camp RJ, Gorresen PM, et al: Collapsing avian tion in response to climate change: projections based on model
community on a Hawaiian island, Sci Adv 2, 2016. analysis, Global Change Biol 20:3845–3858, 2014.
30. Atkinson CT, LaPointe DA: Introduced avian diseases, climate 48. Glibert PM, Anderson DM, Gentien P, et al: The global complex
change, and the future of Hawaiian Honeycreepers, J Avian Med phenomena of harmful algal blooms, Oceanography 18:136–147,
Surg 23:53–63, 2009. 2005.
31. LaPointe DA, Benning TL, Atkinson CT: Avian malaria, climate 49. Kudela R, Pitcher G, Probyn T, et al: Harmful algal blooms in
change and native birds of Hawaii. In Lovejoy TE, Hannah LJ, coastal upwelling systems, Oceanography 18:184–197, 2005.
editors: Climate change and biodiversity, New Haven, CT, 2005, 50. O’Neil J, Davis TW, Burford MA, et al: The rise of harmful
Yale University Press, pp 317–321. cyanobacteria blooms: the potential roles of eutrophication and
32. Atkinson CT, LaPointe DA: Ecology and pathogenicity of avian climate change, Harmful Algae 14:313–334, 2012.
malaria and pox. In Pratt K, Atkinson CT, Banko PC, et al, 51. Burek KA, Gulland FM, O’Hara TM: Effects of climate change
editors: Conservation biology of Hawaiian forest birds: implications on Arctic marine mammal health, Ecol Appl 18:126–134,
for island avifauna, New Haven, CT, 2009, Yale University Press, 2008.
pp 234–252. 52. Van Hemert C, Pearce JM, Handel CM: Wildlife health in
33. Springer YP, Jarnevich CS, Barnett DT, et al: Modeling the a rapidly changing North: focus on avian disease, Front Ecol
present and future geographic distribution of the Lone Star Environ 12:548–556, 2014.
Tick, Amblyomma americanum (Ixodida: Ixodidae), in the 53. Shaman J, Day JF, Stieglitz M: St. Louis encephalitis virus in wild
Continental United States, Am J Trop Med Hyg 93:875–890, birds during the 1990 South Florida epidemic: The importance
2015. of drought, wetting conditions, and the emergence of Culex
34. Maskey JJ, Sweitzer RA, Goodwin BJ: Climate and habitat influ- nigripalpus (Diptera: Culicidae) to arboviral amplification and
ence prevalence of meningeal worm infection in North Dakota, transmission, J Med Entomol 40:547–554, 2003.
USA, J Wildl Dis 51:670–679, 2015. 54. Paull SH, Horton DE, Ashfaq M, et al: Drought and immunity
35. Lenarz MS, Nelson ME, Schrage MW, et al: Temperature medi- determine the intensity of West Nile virus epidemics and climate
ated moose survival in Northeastern Minnesota, J Fish Wildl change impacts, Proc Biol Sci 284:21162078, 2017.
Manag 73:503–510, 2009. 55. George TL, Harrigan RJ, LaManna JA, et al: Persistent impacts
36. Murray DL, Cox EW, Ballard WB, et al: Pathogens, nutritional of West Nile virus on North American bird populations, Proc
deficiency, and climate influences on a declining moose popula- Nat Acad Sci U S A 112:14290–14294, 2015.
tion, Wildlife Monogr 1–29, 2006. 56. Hewson I, Button JB, Gudenkauf BM, et al: Densovirus associ-
37. Laaksonen S, Pusenius J, Kumpula J, et al: Climate change ated with sea-star wasting disease and mass mortality, Proc Nat
promotes the emergence of serious disease outbreaks of filarioid Acad Sci U S A 111:17278–17283, 2014.
nematodes, EcoHealth 7:7–13, 2010. 57. Eisenlord ME, Groner ML, Yoshioka RM, et al: Ochre star
38. Welch DA, Samuel WM, Wilke CJ: Suitability of moose, elk, mortality during the 2014 wasting disease epizootic: role of
mule deer, and white-tailed deer as hosts for winter ticks (Der- population size structure and temperature, Philos Trans R Soc
macentor albipictus), Can J Zool 69:2300–2305, 1991. Lond B Biol Sci 371:2016.
39. Hoar B: Ecology and transmission dynamics of Ostertagia 58. Menge BA, Cerny-Chipman EB, Johnson A, et al: Sea star
gruehneri in barren ground caribou: Dissertation, University of wasting disease in the keystone predator Pisaster ochraceus in
Calgary, 2012. Oregon: Insights into differential population impacts, recovery,
40. Hoegh-Guldberg O, Mumby PJ, Hooten AJ, et al: Coral reefs predation rate, and temperature effects from long-term research,
under rapid climate change and ocean acidification, Science PLoS ONE 11:2016.
318:1737–1742, 2007. 59. Lorch JM, Lankton J, Werner K, et al: Experimental infection
41. Tout J, Siboni N, Messer LF, et al: Increased seawater temperature of snakes with Ophidiomyces ophiodiicola causes pathological
increases the abundance and alters the structure of natural Vibrio changes that typify snake fungal disease, MBio 6:2015.
254 SE C T I O N 8 Emerging and Changing Infectious Diseases
60. Lorch JM, Knowles S, Lankton JS, et al: Snake fungal disease: 65. Friend M, McLean RG, Dein FJ: Disease emergence in birds:
an emerging threat to wild snakes, Philos T R Soc B 371: Challenges for the twenty-first century, Auk 118:290–303, 2001.
2016. 66. Lafferty KD, Porter JW, Ford SE: Are diseases increasing in the
61. Frick WF, Pollock JF, Hicks AC, et al: An emerging disease causes ocean? Annu Rev Ecol Evol Syst 35:31–54, 2004.
regional population collapse of a common North American bat 67. Jones KE, Patel NG, Levy MA, et al: Global trends in emerging
species, Science 329:679–682, 2010. infectious diseases, Nature 451:990–U994, 2008.
62. LaPointe DA, Atkinson CT, Samuel MD: Ecology and conserva- 68. Felton A, Fischer J, Lindenmayer DB, et al: Climate change, con-
tion biology of avian malaria, Ann N Y Acad Sci 1249:211–226, servation and management: an assessment of the peer-reviewed
2012. scientific journal literature, Biodivers Conserv 18:2243–2253,
63. Rosa G, Anza I, Moreira P, et al: Evidence of chytrid-mediated 2009.
population declines in common midwife toad in Serra da Estrela, 69. Walther GR, Post E, Convey P, et al: Ecological responses to
Portugal, Anim Conserv 16:306–315, 2013. recent climate change, Nature 416:389–395, 2002.
64. Pollard AJ, Dobson SR: Emerging infectious diseases in the 21st 70. Bradley CA, Altizer S: Urbanization and the ecology of wildlife
century, Curr Opin Infect Dis 13:265–275, 2000. diseases, Trends Ecol Evol (Amst) 22:95–102, 2007.
37
Prion Diseases in Wildlife
CHRISTINA J. SIGURDSON AND PATRICIA AGUILAR-CALVO
P
rion diseases, also known as transmissible spongiform and atypical BSE in cattle.20–22 For the acquired TSEs, the
encephalopathies (TSEs), are infectious and fatal most plausible route of transmission is through ingestion of
neurodegenerative disorders that include bovine prion-contaminated feed.
spongiform encephalopathy (BSE), TSEs of zoo animals, The efficiency of prion transmission between individuals
scrapie of sheep and goats, transmissible mink encepha- varies widely depending on the prion. BSE in cattle shows
lopathy (TME), and chronic wasting disease (CWD) of little evidence of direct transmission from one individual
cervids (Table 37.1).1–8 To date, outbreaks in zoos have been to another and prions are mostly constrained to the central
limited to small numbers of animals, however, infectious nervous system (CNS). Zoo animals were likely infected
prions can incite large-scale, multispecies epidemics. For through consuming BSE-contaminated meat or meat and
example, within the United Kingdom, BSE infected more bone meal (MBM). In zoo collections, prion infection was
than 180,000 cattle,6,9 as well as zoo bovids, felids, and diagnosed in 19 species, including 8 antelope and bovid
primates.10–16 species [greater kudu (Tragelaphus strepsiceros), eland (Tau-
The infectious agent in prion disease is composed of rotragus oryx), nyala (Tragelaphus angasi), gemsbok (Oryx
the pathogenic misfolded and aggregated prion protein, gazella), scimitar-horned oryx (Oryx dammah), Arabian
known as PrPSc (“Sc” denotes scrapie, the prion disease of oryx (Oryx leucoryx), ankole cows (Bos taurus), and Ameri-
sheep and goats).17,18 The primary amino acid sequence can bison (Bison bison)], 7 felid species [cheetah (Acinonyx
of the prion aggregate is determined by the host cellular jubatus), puma (Felis concolor), ocelot (Felis pardalis), Asian
prion protein, PrPC (“C” denotes the cellular, physiologic golden cat (Catopuma temminckii), leopard cat (Prionailurus
form of the prion protein), encoded by the prion gene, Bengalis), tiger (Panthera tigris), and lion (Panthera leo)],
Prnp.19 Following transmission to the host, prions seed the and 4 nonhuman primates species [Mayotte brown lemurs
misfolding of PrPC in an autocatalytic process. Therefore (Eulemur fulvus), white fronted brown lemur (Eulemur
prions are infectious proteins and do not contain any albifrons), mongoose lemur (Eulemur mongoz), and Rhesus
specific nucleic acids; the term prion is derived from the macaque (Macaca mulatta)] (see Table 37.1).23 Prion disease
words “proteinaceous infectious particle.”18 Over a period outbreaks also occurred in exotic animals in France, Ireland,
of months to years, prions accumulate to high levels in the Germany, and even Australia, although almost all of the
brain and spinal cord, resulting in spongiform degeneration infected animals originated from zoological collections in
with vacuolation, neuronal death, and activated astrocytes the United Kingdom.1
and microglia. Although the incubation period can be In contrast to BSE in cattle, scrapie and CWD are highly
many years, even decades, the clinical phase is typically transmissible between animals. In scrapie-infected sheep
rapidly progressive (weeks to months) and may include and CWD-infected deer, prions accumulate in lymphoid
behavior abnormalities, motor dysfunction, cognitive tissues, including tonsils, lymph nodes, and Peyer patches
impairment, and ataxia, depending on the prion and species (Fig. 37.1).24–26 CWD prions are shed in the saliva, urine,
affected. and feces directly into the environment,27,28 contaminating
Here we review the transmission and epidemiology, clini- grazing areas and water sources. CWD prions can be trans-
cal signs and diagnosis, and the treatment and prevention mitted via fomites; for example, feed buckets and bedding
of prion diseases in zoo animals and in free-ranging cervids. used by CWD-infected deer can transmit the infection
to uninfected deer.29 Additionally, direct animal-to-animal
Transmission and Epidemiology contact or contact with prion-contaminated secretions and
excretions, such as feces, urine, saliva, blood, placenta,
Most prion diseases of animals are acquired by exposure to and milk, are possible routes of infection.30–32 Vertical
prions, however prions can also arise sporadically in aged transmission to offspring has also been demonstrated for
animals, for example, atypical scrapie in sheep and goats prion-infected sheep, goats, and deer.33,34
255
256 SE C T I O N 8 Emerging and Changing Infectious Diseases
TABLE
37.1 Prion Diseases in Wild and Captive Animals
Year of
Disease Host Etiology Description
Transmissible mink encephalopathy Mink Ingestion of bovine spongiform 19653,4
(TME) encephalopathy (BSE) or scrapie-
contaminated meat and bone meal
Feline spongiform encephalopathy Domestic cat, cheetah, ocelot, Ingestion of BSE-contaminated meat 19901,5
(FSE) puma, lion, Asian golden and bone meal
cat, leopard cat, tiger
Exotic ungulate spongiform Kudu, eland, gemsbok, nyala, 19892
encephalopathy oryx, bison
Transmissible spongiform Lemur, rhesus macaque 199611
encephalopathy in nonhuman
primates
Chronic wasting disease (CWD) Mule deer, white-tailed deer, Unknown source; highly transmissible 19677,65
moose, reindeer, sika deer, through the ingestion or direct
Rocky Mountain elk contact with pastures, soil, feces,
urine, saliva, or blood
Bovine spongiform encephalopathy Cattle Ingestion of BSE-contaminated meat 19866
(BSE) and bone meal
Bovine amyloidotic spongiform Cattle Unknown source; probably 200466
encephalopathy (BSE-L) spontaneous origin
Atypical spongiform Cattle Unknown source; probably 200467
encephalopathy Type-H (BSE-H) spontaneous origin
Scrapie Sheep, goat, mouflon Unknown source; transmissible 173268
through the ingestion or direct
contact with pastures, feces,
placenta, or blood
Atypical scrapie Sheep and goat Unknown source; probably 199869
spontaneous origin
A B C
• Figure 37.1 Brainstem and tonsil sections from a chronic wasting disease-infected mule deer. (A)
Hematoxylin and eosin stain of brain shows neurons and adjacent vacuoles (arrows), lesions typical of a
spongiform encephalopathy. (B) Immunohistochemical (IHC) stain for PrP shows abundant PrPSc deposits
(red), here shown on the neuronal plasma membrane (center). (C) IHC of the tonsil reveals PrPSc deposits
in the germinal center of lymphoid follicles. Scale bars = 50 µm (A), 100 µm (B), or 500 µm (C).
a slow disease progression over weeks is common. The main were diagnosed with CWD. For the next two decades,
signs reported include ataxia, tremor, abnormal head and CWD was thought to exist exclusively in Colorado and
ear posture, and weight loss. Myoclonus, dullness, excessive Wyoming. In 1995, BSE prions were discovered to have
movements of the lips and tongue, nibbling of the tail, spread from cattle to humans, and with a zoonotic risk of
decreased rumination, hyperesthesia, and anxiety may also prions recognized, CWD surveillance surged throughout
be observed.1,38 North America and revealed isolated, noncontiguous popu-
The incubation period for feline spongiform encepha- lations of CWD-infected cervids from Utah to New York,
lopathy (FSE) in cheetahs ranges from 4.5 to 8 years. For and south to Texas and Arkansas, as well as in two Canadian
domestic cats, the incubation period is unknown, although provinces (Fig. 37.2). Highly reported prevalences in certain
all affected animals were at least 2 years old. Clinical signs states, including Wisconsin and Arkansas (>20%), suggests
rapidly progress over 1–3 months.23 Domestic cats initially that CWD has been established in some regions for more
display behavior changes such as unusual timidity, hiding, than a decade. To date, CWD-infected cervids have been
and aggressiveness, followed by ataxia, gait abnormalities reported in 25 US states, 2 Canadian provinces, South
(mainly affecting the hind legs), hypermetria, and hyper- Korea (ranched elk imported from North America), and
esthesia to sound and touch. Decreased grooming, exces- most recently in Europe. In 2016, free-ranging reindeer
sive salivation, polyphagia, polydipsia, and dilated pupils and moose in Norway were diagnosed with CWD,44 raising
have also been reported. In later stages of the disease, questions of how CWD prions spread to free-ranging
animals are somnolent and may develop tremors. Similar reindeer and moose populations in northern Europe.
behavior and motor signs have been described in zoo cat Possibilities include spread by commercially available
species.39 cervid urine used as a hunting bait, CWD-contaminated
Prion-infected nonhuman primates initially show behav- fomites, or horizontal spread from sporadic prion
ioral signs of depression, nervousness, voracious appetite, disease.
teeth grinding, yawning, and altered grooming activity, Is interspecies prion transmission a risk, given that
which progresses to truncal ataxia with tremors. In end predators, scavengers, rodents, ungulates, and birds are
stages of disease, animals display myoclonic jerks, ataxia, likely exposed to CWD-infected cervids? There are no
and aggressiveness.23,40 known naturally occurring cases of CWD in noncervid
In TME-infected mink, the incubation period ranges species, suggesting a high transmission barrier, yet cross-
from 7 to 12 months. Progression of the disease is similar species transmission remains a possibility.
to FSE, as mink show altered behavior with hyperesthesia,
increased aggressiveness, and hyper-irritability, followed by Clinical Disease
ataxia, circling, tremors, and compulsive biting of self or
objects. Death occurs one week to several months after The initial stages of CWD are clinically subtle and include
disease onset.23,41 weight loss and behavior changes, such as depression
and isolation from the herd. Animals often appear thin
Chronic Wasting Disease and unthrifty, and may develop polyuria and polydipsia,
hypersalivation, ataxia, frequent regurgitation, and esopha-
CWD is among the most infectious prion disease in animals, geal distension.42 The cause of death is commonly due to
affecting captive and free-ranging Cervidae including mule aspiration pneumonia. Given that the clinical signs are
deer (Odocoileus hemionus), white-tailed deer (O. virgin- often subtle and nonspecific, surveying cervid populations
ianus), Rocky Mountain elk (Cervus canadensis nelsoni), is critical to avoid large-scale epidemics within a collec-
moose (Alces alces), sika deer (Cervus nippon), and reindeer tion and to prevent transmission of CWD among cervid
(Rangifer tarandus tarandus). Reported prevalences of up populations.
to 35% in free-ranging animals and 90% in captivity are
a testament to the efficient transmission of CWD prions. Transmission
Importantly, CWD prions can cross species barriers and
infect other cervids, including Reeves’ muntjac (Muntia- CWD prions are not restricted to the CNS and also accu-
cus reevesi). No cervid species should be assumed to be mulate in systemic organs beginning in early disease. Few
CWD-resistant. As CWD prions are typically shed into the other prion infections induce such extensive deposits of
environment, outbreaks of CWD are best identified early extraneural prions. In CWD-infected deer, prion aggregates
and controlled expeditiously. Two CWD-positive animals accumulate in all lymphoid tissues including tonsils (see
have been reported in zoological collections.42 Fig. 37.1), lymph nodes, Peyer patches, and spleen, as well
as in adrenal glands, pancreatic islets, pituitary, and skeletal
Epidemiology muscle.25,26 Such an extensive extraneural reservoir may
underlie the efficient horizontal spread of CWD prions.
CWD was initially discovered in captive mule deer in Prions are shed in saliva, urine, and feces, and these excreta,
northern Colorado in 1967.43 Soon thereafter, captive Rocky as well as blood, can transmit CWD to naïve animals.27,28
Mountain elk in Wyoming as well as free-ranging cervids Shed prions bound to soil contaminate the environment,
258 SE C T I O N 8 Emerging and Changing Infectious Diseases
and exposure to prion-contaminated fomites, that is, water aggregated isoform, PrPSc, from the normal cellular isoform,
sources or bedding, can initiate an infection. PrPC, as PrPSc is relatively resistant to protease digestion
and is detected with anti-PrP antibodies. ELISA-based
Diagnostic Tools for Prion Disease diagnostic kits are available for testing brain samples.
Histologic examination of the brain typically reveals
Extraneural prion accumulation presents opportuni- neuronal vacuolation and astrogliosis (see Fig. 37.1). Each
ties for antemortem screening of cervid populations. prion disease produces a different disease phenotype in
In deer, prion immunolabelling of recto-anal mucosa- terms of brain areas affected, size and morphology of prion
associated lymphoid tissue biopsies are a sensitive and spe- plaques, and level of gliosis and vacuolation. For example,
cific method for testing live animals for CWD infection.45,46 FSE is characterized by spongiform degeneration in the
CWD-infected elk cases may be missed, however, as not all neuropil of the brain and spinal cord with the most severe
elk develop prions in lymphoid tissues. Similarly, scrapie lesions localized to the medial geniculate nucleus of the
can be detected by prion immunolabelling of the third thalamus as well as the basal nuclei. TME-infected mink
eyelid lymphoid follicles, although this test is only highly develop extensive neuropil vacuolation, as well as neuronal
sensitive by 10–14 months or later after infection.47 degeneration and astrocytosis in the cerebral cortex, particu-
Postmortem tests for prion disease are numerous and larly in the frontal cortex, as well as the corpus callosum.
include immunohistochemical staining of brainstem, or New extraordinarily sensitive techniques are under
western blot, or commercial ELISA assays to detect aggre- development for large-scale population testing of body
gated prions. Prion diseases are diagnosed postmortem by fluids or tissue samples. Protein misfolding cyclic amplifica-
detection of PrPSc aggregates in the CNS with immunob- tion (PMCA) and real-time quaking induced conversion
lotting, ELISA, or IHC (see Fig. 37.1). These diagnostic (RT-QuIC) assays amplify low levels of aggregated prions in
methods allow the discrimination of the pathologic early stages of the disease with high sensitivity in a number
CHAPTER 37 Prion Diseases in Wildlife 259
of tissues and body fluids, including cerebrospinal fluid information sheet is available from the American Associa-
(CSF), urine, saliva, blood, brain tissue, and lymph node tion of Zoo Veterinarians Infectious Disease Committee at
tissue.48–51 http://www.aazv.org/?754.
19. Kretzschmar HA, Stowring LE, Westaway D, et al: Molecular spongiform encephalopathy in a German captive cheetah, J Gen
cloning of a human prion protein cDNA, DNA 5:315–324, Virol 91:2874–2883, 2010.
1986. 40. Bons N, Mestre Frances N, Charnay Y, et al: Spontaneous
20. Baron T, Biacabe AG, Arsac JN, et al: Atypical transmissible spongiform encephalopathy in a young adult rhesus monkey, C
spongiform encephalopathies (TSEs) in ruminants, Vaccine R Acad Sci III 319:733–736, 1996.
25:5625–5630, 2007. 41. Schneider DA, Harrington RD, Zhuang D, et al: Disease-
21. Benestad SL, Arsac JN, Goldmann W, et al: Atypical/Nor98 associated prion protein in neural and lymphoid tissues of mink
scrapie: properties of the agent, genetics, and epidemiology, Vet (Mustela vison) inoculated with transmissible mink encephalopa-
Res 39:19, 2008. thy, J Comp Pathol 147:508–521, 2012.
22. Biacabe AG, Morignat E, Vulin J, et al: Atypical bovine spon- 42. Williams ES, Miller MW: Chronic wasting disease in deer and
giform encephalopathies, France, 2001–2007, Emerg Infect Dis elk in North America, Rev - Off Int Epizoot 21:305–316, 2002.
14:298–300, 2008. 43. Williams ES, Young S: Spongiform encephalopathies in Cervi-
23. Sigurdson CJ, Miller MW: Other animal prion diseases, Br Med dae, Rev - Off Int Epizoot 11:551–567, 1992.
Bull 66:199–212, 2003. 44. Benestad SL, Mitchell G, Simmons M, et al: First case of chronic
24. Hadlow WJ, Kennedy RC, Race RE: Natural infection of Suffolk wasting disease in Europe in a Norwegian free-ranging reindeer,
sheep with scrapie virus, J Infect Dis 146:657–664, 1982. Vet Res 47:88, 2016.
25. Sigurdson CJ, Spraker TR, Miller MW, et al: PrP(CWD) in the 45. Wild MA, Spraker TR, Sigurdson CJ, et al: Preclinical diagnosis
myenteric plexus, vagosympathetic trunk and endocrine glands of chronic wasting disease in captive mule deer (Odocoileus
of deer with chronic wasting disease, J Gen Virol 82:2327–2334, hemionus) and white-tailed deer (Odocoileus virginianus) using
2001. tonsillar biopsy, J Gen Virol 83:2629–2634, 2002.
26. Sigurdson CJ, Williams ES, Miller MW, et al: Oral transmis- 46. Keane D, Barr D, Osborn R, et al: Validation of use of rectoanal
sion and early lymphoid tropism of chronic wasting disease mucosa-associated lymphoid tissue for immunohistochemical
PrPres in mule deer fawns (Odocoileus hemionus), J Gen Virol diagnosis of chronic wasting disease in white-tailed deer (Odocoi-
80:2757–2764, 1999. leus virginianus), J Clin Microbiol 47:1412–1417, 2009.
27. Mathiason CK, Powers JG, Dahmes SJ, et al: Infectious prions in 47. O’Rourke KI, Baszler TV, Besser TE, et al: Preclinical diagnosis
the saliva and blood of deer with chronic wasting disease, Science of scrapie by immunohistochemistry of third eyelid lymphoid
314:133–136, 2006. tissue, J Clin Microbiol 38:3254–3259, 2000.
28. Tamguney G, Miller MW, Wolfe LL, et al: Asymptomatic deer 48. Saborio GP, Permanne B, Soto C: Sensitive detection of patho-
excrete infectious prions in faeces, Nature 461:529–532, 2009. logical prion protein by cyclic amplification of protein misfold-
29. Mathiason CK, Hays SA, Powers J, et al: Infectious prions in ing, Nature 411:810–813, 2001.
pre-clinical deer and transmission of chronic wasting disease 49. Wilham JM, Orru CD, Bessen RA, et al: Rapid end-point
solely by environmental exposure, PLoS ONE 4:e5916, 2009. quantitation of prion seeding activity with sensitivity comparable
30. Scott JR: Scrapie pathogenesis, Br Med Bull 49:778–791, to bioassays, PLoS Pathog 6:e1001217, 2010.
1993. 50. Atarashi R, Satoh K, Sano K, et al: Ultrasensitive human prion
31. Taylor DM, McConnell I, Fraser H: Scrapie infection can be detection in cerebrospinal fluid by real-time quaking-induced
established readily through skin scarification in immunocompe- conversion, Nat Med 17:175–178, 2011.
tent but not immunodeficient mice, J Gen Virol 77:1595–1599, 51. Orru CD, Bongianni M, Tonoli G, et al: A test for Creutzfeldt-
1996. Jakob disease using nasal brushings, N Engl J Med 371:519–529,
32. Mohan J, Brown KL, Farquhar CF, et al: Scrapie transmission 2014.
following exposure through the skin is dependent on follicular 52. Ehlers B, Diringer H: Dextran sulphate 500 delays and prevents
dendritic cells in lymphoid tissues, J Dermatol Sci 35:101–111, mouse scrapie by impairment of agent replication in spleen, J
2004. Gen Virol 65:1325–1330, 1984.
33. Gough KC, Maddison BC: Prion transmission: prion excretion 53. Farquhar CF, Dickinson AG: Prolongation of scrapie incubation
and occurrence in the environment, Prion 4:275–282, 2010. period by an injection of dextran sulphate 500 within the month
34. Nalls AV, McNulty E, Powers J, et al: Mother to offspring before or after infection, J Gen Virol 67:463–473, 1986.
transmission of chronic wasting disease in reeves’ muntjac deer, 54. Diringer H, Ehlers B: Chemoprophylaxis of scrapie in mice, J
PLoS ONE 8:e71844, 2013. Gen Virol 72:457–460, 1991.
35. Marsh RF, Hadlow WJ: Transmissible mink encephalopathy, 55. Kimberlin RH, Walker CA: Antiviral compound effective against
Rev - Off Int Epizoot 11:539–550, 1992. experimental scrapie [letter], Lancet 2:591–592, 1979.
36. Barlow RM: Transmissible mink encephalopathy: pathogenesis 56. Kimberlin RH, Walker CA: The antiviral compound HPA-23
and nature of the aetiological agent, J Clin Pathol Suppl (R Coll can prevent scrapie when administered at the time of infection,
Pathol) 6:102–109, 1972. Arch Virol 78:9–18, 1983.
37. Baron T, Bencsik A, Biacabe AG, et al: Phenotypic similarity of 57. Beringue V, Adjou KT, Lamoury F, et al: Opposite effects of
transmissible mink encephalopathy in cattle and L-type bovine dextran sulfate 500, the polyene antibiotic MS-8209, and congo
spongiform encephalopathy in a mouse model, Emerg Infect Dis red on accumulation of the protease-resistant isoform of PrP in
13:1887–1894, 2007. the spleens of mice inoculated intraperitoneally with the scrapie
38. Kirkwood JK, Cunningham AA, Wells GA, et al: Spongiform agent, J Virol 74:5432–5440, 2000.
encephalopathy in a herd of greater kudu (Tragelaphus strep- 58. Nodelijk G, van Roermund HJ, van Keulen LJ, et al: Breeding
siceros): epidemiological observations, Vet Rec 133:360–364, with resistant rams leads to rapid control of classical scrapie in
1993. affected sheep flocks, Vet Res 42:5, 2011.
39. Eiden M, Hoffmann C, Balkema-Buschmann A, et al: Bio- 59. Kimberlin RH: Aetiology and genetic control of natural scrapie,
chemical and immunohistochemical characterization of feline Nature 278:303–304, 1979.
CHAPTER 37 Prion Diseases in Wildlife 261
60. Hagenaars TJ: Control of scrapie by selective breeding: what are 65. Williams ES, Young S: Spongiform encephalopathy of Rocky
we getting for free?, Vet Rec 174:528–529, 2014. Mountain elk, J Wildl Dis 18:465–471, 1982.
61. Kanata E, Humphreys-Panagiotidis C, Giadinis ND, et al: 66. Casalone C, Zanusso G, Acutis P, et al: Identification of a second
Perspectives of a scrapie resistance breeding scheme targeting bovine amyloidotic spongiform encephalopathy: molecular simi-
Q211, S146 and K222 caprine PRNP alleles in Greek goats, Vet larities with sporadic Creutzfeldt-Jakob disease, Proc Natl Acad
Res 45:43, 2014. Sci USA 101:3065–3070, 2004.
62. Lacroux C, Perrin-Chauvineau C, Corbiere F, et al: Genetic 67. Biacabe AG, Laplanche JL, Ryder S, et al: Distinct molecular
resistance to scrapie infection in experimentally challenged goats, phenotypes in bovine prion diseases, EMBO Rep 5:110–115,
J Virol 88:2406–2413, 2014. 2004.
63. Aguilar-Calvo P, Garcia C, Espinosa JC, et al: Prion and prion- 68. Plummer PJ: Scrapie—a disease of sheep: a review of the litera-
like diseases in animals, Virus Res 207:82–93, 2015. ture, Can J Comp Med Vet Sci 10:49–54, 1946.
64. Fox KA, Jewell JE, Williams ES, et al: Patterns of PrPCWD 69. Benestad SL, Sarradin P, Thu B, et al: Cases of scrapie with
accumulation during the course of chronic wasting disease infec- unusual features in Norway and designation of a new type,
tion in orally inoculated mule deer (Odocoileus hemionus), J Gen Nor98, Vet Rec 153:202–208, 2003.
Virol 87:3451–3461, 2006.
38
Avian Influenza: A Brief Overview of
the Pathobiology and Current Status
in Domestic and Nondomestic Species
DARREL K. STYLES AND YVONNE NADLER
262
CHAPTER 38 Avian Influenza: A Brief Overview of the Pathobiology and Current Status in Domestic and Nondomestic Species 263
tissue tropism results because of the presence of HA-specific manifesting severe clinical illness themselves. In the past
enzymes needed to cleave the hemagglutinin protein (an two decades, HPAI H5N1 has continued to diversify, and
essential step for viral infection of the cell) after attachment the lineage has given rise to multiple sublineages designated
to the cell in the avian intestinal or respiratory tract. As in phylogenetic terms as clades.
alluded to earlier, an important cell tropism and patho-
genicity determinant of AIVs is the sequence and types The New Paradigm
of amino acids associated with the HA cleavage site. If a
single basic amino acid is strategically located at the cleavage The HPAI H5 clade 2.3.4.4 of avian influenza (first reported
point within HA, this restricts the enzyme or enzymes (and in domestic ducks in China in 2008) burst forth from Asia
thereby the cell types) that may successfully cleave the HA in 2013–2015 to create an intermittent global epiornitic,
protein to allow fusion with the host cell membrane and even reaching North America in 2014–2015.15 This clade
completion of the replication cycle. LPAI viruses possess a of H5 viruses achieved what their progenitor HPAI H5N1
single basic amino acid at the cleavage point within their could not—namely global distribution—and it appears to
HA protein; this permits action only by enzymes found in be uniquely adapted to the dabbling duck (genus Anas).
the intestinal tract of waterfowl. By contrast, HPAI viruses H5 clade 2.3.4.4 includes multiple subtypes, but the most
have multiple basic amino acids located in the cleavage notable are Eurasian lineage HPAI H5N8, H5N6, and
region; this permits an array of enzymes of multiple cell H5N2.16 Despite descending from the zoonotic HPAI
types to actively cleave the HA protein. Therefore HPAI H5N1, few of these novel subtypes have demonstrated the
viruses are systemic in nature and may infect multiple organ same zoonotic potential with the exception of EA HPAI
systems, whereas LPAI viruses are limited to single organ H5N6.
systems. Eurasian HPAI H5N8 (EA HPAI H5N8) has achieved
To meet the regulatory definition for HPAI, the viral a global distribution through movement by wild migratory
HA must first be sequenced to ascertain the composition waterfowl, whereas H5N6 and H5N2 have largely been
of the cleavage region. If multiple basic amino acids are limited to Asia to date. This clade is now well established
detected in this region, the virus by definition is considered throughout Asia and possibly other areas. H5 clade 2.3.4.4
to be HPAI irrespective of its clinical manifestation. Most viruses do not appear to cause clinical disease in many
HPAI viruses are indeed highly pathogenic, but some may species of wild or domestic dabbling ducks but have been
be subclinical in behavior depending on the species and shown to be lethal to other wild bird species such as raptors,
inherent characteristics of the virus.12 geese, and swans.
Beginning in January 2014, H5N8 spread rapidly
through domestic poultry operations in South Korea and
The Changing Ecology of Avian even resulted in mass mortalities of some wild birds such as
Influenza Viruses Baikal teal (Anas formosa). Then, in the fall of 2014, H5N8
reached the Middle East and Europe. By November 2014,
For many years, the long-held dogma was that low- H5N8 was detected in North America and presumably
pathogenicity viruses were found in wild waterfowl and arrived through viral exchange in Beringia between infected
occasionally spilled over into the domestic poultry com- Eurasian ducks and North American ducks.17 H5N8 reas-
partment but that highly pathogenic viruses were almost sorted with an endemic North American LPAI virus to give
exclusively limited to domestic poultry, with very rare rise to novel reassortants including Eurasian/North Ameri-
exceptions.13 However, a novel AIV emerged in Asia that can HPAI H5N2 and HPAI H5N1 (EA/NA HPAI H5N2
was to change that paradigm. The first detection of Asian and H5N1). Both H5N8 and H5N2 were carried south by
lineage HPAI H5N1 (HPAI H5N1) was in 1997 in Hong wild migratory waterfowl, predominantly dabbling ducks.
Kong, where it caused severe disease in domestic poultry There was only a single detection of EA/NA HPAI H5N1,
and exhibited a zoonotic potential resulting in illness and and it did not appear to persist or spread in the waterfowl
death in humans.14 Rapid response to the outbreak seemed population. However, H5N8 did spread in wild ducks and
to eradicate the virus, but HPAI H5N1 did not disap- precipitated outbreaks in domestic poultry in the United
pear; in fact, it continued to persist and reassort in wild States, largely in the fall of 2014 and early winter of 2015
migratory waterfowl and domestic poultry. HPAI H5N1 along the Pacific flyway. H5N2 caused serious outbreaks
spread throughout East and Southeast Asia and became in domestic turkeys in British Columbia, Canada, in late
endemic in that region. However, in 2005, the virus was 2014 but did not result in significant outbreaks in the
carried by wild migratory waterfowl into the Middle East, United States until the spring of 2015. Starting in March
Europe, and Africa.4 This was the first HPAI virus that 2015 and coinciding with the northern migration in the
appeared to successfully infect wild and domestic waterfowl Mississippi flyway, H5N2 precipitated the largest cluster of
(predominantly in the genus Anas) without causing signifi- HPAI outbreaks in domestic poultry in the history of the
cant clinical disease in all cases. Hence, these birds could United States, mainly in the central and upper Midwest.
move HPAI H5N1 long distances along migration routes These outbreaks continued and were not resolved until
and precipitate outbreaks in domestic poultry without June 2015.
CHAPTER 38 Avian Influenza: A Brief Overview of the Pathobiology and Current Status in Domestic and Nondomestic Species 265
Wild bird surveillance has shown that neither H5N8 concepts and to translate them into site-specific actions
nor H5N2 appeared to persist long-term in the wildlife depending on risk of infection. First, conceptual biosecurity
compartment, with only two geographically independent should include a site evaluation relative to surrounding
detections in single mallards (Anas platyrhynchos) in the risk factors such as wetlands (HPAI reservoir habitat) or
summer and fall of 2015 and a single detection in a poultry operations. Structural biosecurity measures are the
mallard in the summer of 2016. Similar observations have “physical” barriers to disease. In the 2014–2015 HPAI
been made after incursions into Europe from 2013 to the outbreaks in the Midwest, specimens in high-value collec-
present. This suggests that H5 clade 2.3.4.4 does not readily tions were housed indoors until the disease threat had been
persist within the wild migratory waterfowl compartment minimized. Other facilities were able to install temporary
at prevalence rates detectable by the various surveillance “covers” to minimize the entry to certain exhibits by wild
systems in place globally. This notion is supported by the waterfowl. Operational biosecurity should describe how the
fact that outbreaks in domestic poultry diminish and then site is managed to prevent the introduction of pathogens.
disappear outside of the migration season. Moreover, active Personnel protocols, personal protective equipment (PPE),
surveillance within the US wild bird compartment shows and procedures for movement, cleaning, and disinfection of
that these H5 viruses circulated at a low prevalence in that equipment and how these materials are moved through the
population. Hence, one might speculate that to maintain facility are elements of operational biosecurity. Examples
infection of the wild waterfowl compartment with subtypes include changing footwear when entering animal enclosures
of this clade of H5 viruses requires frequent spillover events and ensuring that any free-roaming fowl (e.g., peafowl and
from infected domestic poultry. Indeed, domestic ducks pheasants) may be caged when there is a threat.
have been shown to be important subclinical reservoirs Vaccination of a collection is a last resort, and any threat
for Eurasian H5 HPAI viruses.18 Additionally, there is from infection must be severe to warrant implementing this
evidence suggesting that residual cross-protection generated countermeasure. There are many impediments to vaccinat-
from natural infection by endemic LPAI viruses provides ing nondomestic species. First, any such procedure must be
some heterosubtypic immunity, which may be another cleared with the national and state/provincial animal health
factor contributing to the failure of EA H5 HPAI clades authorities because there are many regulations surrounding
to persist in wild waterfowl populations.19 Last, one might the use of avian influenza vaccines directed against notifi-
hypothesize that EA H5 HPAI viruses are less virally “fit” in able subtypes. Second, the types of vaccines available for
wild waterfowl as compared with LPAI strains that naturally use in nondomestic birds is largely limited to inactivated
circulate in this population. Therefore the LPAI viruses are products, because the efficacy of modern vectored or modi-
more competitive in the host. fied live-vectored vaccine products (those routinely used in
It is essential to recognize that as this clade of EA H5 domestic poultry) is largely unknown and such products
HPAI viruses maintains wide circulation in Asia, it will may not be efficacious for nondomestic avian species. Third,
continue to evolve and pose an ongoing threat to domestic the performance of even inactivated products may be highly
poultry and wild birds globally. Whether and when this variable across species lines in terms of efficacy and duration
clade of H5 viruses will return via migration to either of immunity. Lastly, vaccinated birds may have to be appro-
Europe or North America depends on a number of factors priately identified by some type of permanent marker (e.g.,
that are not well defined but may include weather patterns a tag, microchip), and the movement of vaccinated birds
and storms affecting migration and the prevalence of infec- may well be restricted both domestically and internationally
tion in domestic poultry (especially domestic ducks of the by trade law owing to the risk of imperiling commerce in
genus Anas). poultry products. Nevertheless, vaccination of nondomestic
avian species has been accomplished successfully in coun-
tries that are under constant or intermittent threat from
Biosecurity of Collections and Vaccination HPAI.21,22 It is important to recognize that vaccination is
of Nondomestic Avian Species a tool and that it should not supplant any of the tenets of
biosecurity.
Zoos and exhibitors face a number of issues when trying
to safeguard collections against incursions from potentially Summary
infected wild migratory waterfowl, and zoos have been
affected by HPAI H5N8 clade 2.3.4.4.20 The water features Avian influenza evolves in unpredictable ways that will
present in many zoological parks are attractive to wildlife continue to challenge the health of both wild and domestic
and serve as a ready stopover point, replete with food and species. The dynamics of viral exchange between recognized
shelter, for migrants. This makes management of outdoor host species such as ducks and land-based poultry may con-
collections challenging. It is probably best for zoos to tribute to the ongoing genetic destabilization of the virus
consult experts on wildlife deterrents; these can be found and increases in virulence. The recently acquired ability of
in many state or federal wildlife agencies. However, there these novel influenza viruses to be subclinically carried and
are basic precautions that may be taken to minimize risk of transmitted by select species of wild waterfowl increases
infection to collections. It is helpful to consider biosecurity the threat to captive collections globally. However, the best
266 SE C T I O N 8 Emerging and Changing Infectious Diseases
tools to defend zoological parks and collections against this 12. Capua I, Alexander DJ: Avian influenza infection in birds: a
continuing threat are vigilance and practical biosecurity. challenge and opportunity for the poultry veterinarian, Poult Sci
88(4):842–846, 2009.
13. Vandergrift K, Sokolow S, Kilpatrick A: Ecology of avian influenza
References viruses in a changing world, Ann N Y Acad Sci 1195:113–128,
2010.
1. Gilbert M, Philippa J: Avian influenza H5N1 virus: epidemiol- 14. Li K, Guan Y, Wang J, et al: Genesis of a highly pathogenic
ogy in wild birds, zoo outbreaks and zoo vaccination policy. and potentially pandemic H5N1 virus in eastern Asia, Nature
In Fowler’s zoo and wild animal medicine current therapy (vol 7). 430(6996):209–213, 2004.
St. Louis, MO, 2012, Elsevier, pp 343–348. 15. Taubenberger J, Morens D: H5Nx panzootic bird flu – influ-
2. Samji T, Influenza A: Understanding the viral life cycle, Yale J enza’s newest worldwide evolutionary tour, Emerg Infect Dis
Biol Med 82(4):153–159, 2009. 23(2):340–342, 2017.
3. Zambon MC: Epidemiology and pathogenesis of influenza, J 16. De Vries E, Hongbo G, Meiling D, et al: Rapid emergence of
Antimicrob Chemother 44(suppl B):3–9, 1999. highly pathogenic avian influenza subtypes from a subtype of
4. Gilbert M, Xiangming X, et al: Anatidae migration in the western H5N1 hemagglutinin variant, Emerg Infect Dis 21(5):842–846,
Palearctic and spread of highly pathogenic avian influenza virus 2015.
H5N1, Emerg Infect Dis 12(11):1650–1656, 2006. 17. Lee D, Torchetti M, Winker K, et al: Intercontinental spread
5. Tong S, Li Y, Rivailler P, et al: A distinct lineage of influenza A of Asian-origin H5N8 to North America through Beringia by
virus from bats, Proc Natl Acad Sci USA 109(11):4269–4274, migratory birds, J Virol 89(12):6321–6324, 2015.
2012. 18. Song Y, Cul J, Hui S, et al: New reassortant H5N8 highly
6. Tong S, Zhu X, Li Y, et al: New world bats harbor diverse pathogenic avian influenza virus from waterfowl in Southern
influenza A viruses, PLoS Pathog 9(10):e1003657, 2013. China, Frontiers Micro 6:2015.
7. Bodewes R, Morick D, et al: Recurring influenza B viruses in 19. Latorre-Margalef N, Brown J, Fojtik A, et al: Competition
seals, Emerg Infect Diseases 19(3):511–512, 2013. between influenza A virus subtypes through heterosubtypic
8. Hause B, Collin E, et al: Characterization of a novel influenza immunity modulates re-infection and antibody dynamics in the
virus in cattle and swine: proposal for a new genus in the Ortho- mallard duck, PLoS Pathog 13(6):2017.
myxoviridae family, MBio 5(2):1–10, 2014. 20. Hagarajan S, Kumar M, Murugkar H, et al: Novel reassortant
9. Taubenberger JK, Kash JC: Influenza virus evolution, host adap- highly pathogenic avian influenza (H5N8) virus in zoos, India,
tation and pandemic formation, Cell Host Microbe 7(6):440–451, Emerg Infect Dis 23(4):717–719, 2017.
2010. 21. Lécu A, De Langhe C, Petit T, et al: Serologic response and
10. Thontiravong A, Kitikoon P, Wannaaratana S: Quail as a poten- safety to vaccination against avian influenza using inactivated
tial mixing vessel for the generation of new reassortant influenza H5N2 vaccine in zoo birds. J Zoo Wildl Med 40(4):731–743,
A viruses, Vet Micro 160:305–313, 2012. 2009.
11. Maurer-Stroh S, Lee R, Gunalan V, et al: The highly pathogenic 22. Lierz M, Hafez H, Klopfleisch R, et al: Protection and virus
H7N3 avian influenza strain from 2012 in Mexico acquired an shedding of falcons vaccinated against highly pathogenic avian
extended cleavage site through recombination with host 28S influenza A virus (H5N1), Emerg Infect Dis 13(11):1667–1674,
rRNA, Virol J 10(1):2013. 2007.
39
Emerging Reptile Viruses
RACHEL E. MARSCHANG
I
n reptile virology, it may be difficult to recognize the and squamates as well as in amphibians and fish; they are
true emergence of a pathogen, as detection of previously covered in Chapter 52.
unknown organisms has occurred rapidly in recent years.
This has been due both to the use of new technologies in Adenoviridae
reptile diagnostics and to the increased interest of research-
ers from various backgrounds in reptiles as virus hosts. Adenoviruses are large, nonenveloped double-stranded (ds)
The increasing use of next-generation sequencing (NGS) DNA viruses. Members of three different genera have been
methods has led to many of these discoveries and appears described in reptiles: atadenoviruses, siadenoviruses, and
likely to increase our knowledge of viruses in zoo, aquarium, the proposed “testadenoviruses.”6 The atadenoviruses have
and wildlife species significantly in the coming years. The been hypothesized to have evolved in squamate reptiles,7
development and commercial availability of sensitive diag- and some of the viruses found in these reptiles appear to be
nostic tests has increased, and changes in detection rates host species–specific. Atadenovirus infections in squamates
may not always reflect a true increase in viral prevalence are not always associated with disease, although they may
among reptiles. This underlines the importance of additional be involved in multipathogen disease processes. There is
studies using a wide range of disciplines and techniques to increasing evidence that numerous squamate atadenoviruses
understand both the clinical significance of many viruses are able to switch between various squamate hosts, with
in various reptile species as well as their host ranges, the unknown clinical and epidemiologic consequences8 as well
pathogenesis of associated disease, and epidemiology. as ramifications for viral detection methods. Although
Numerous factors influence the emergence of viral atadenoviral infections in squamates are well documented,
infections in reptile populations. These include effects of reports of adenoviral infections in chelonians are relatively
climate change, which may influence the spread of viral rare and the genetic diversity of the detected viruses is
infections in reptiles in multiple ways, including increasing relatively high, indicating that at least in some cases adeno-
the range of invertebrate vectors, especially for arboviruses viruses have recently switched to chelonians as hosts. The
(Table 39.1); influencing the reptile immune system; and first report of a disease outbreak associated with adeno-
the replication rates of viruses capable of infecting reptiles. viral infection in chelonians was in a group of Sulawesi
Direct human interaction has been shown to affect the tortoises (Indotestudo forsteni) that were illegally imported
spread of multiple diseases in wild animals, and the pet into the United States.9 Affected animals developed severe
trade plays a large role in the international spread of viruses multisystemic disease with clinical signs including anorexia,
(e.g., for ranaviruses,1,2 herpesviruses,3 and reptarenaviruses) lethargy, mucosal ulcerations, nasal and ocular discharge,
in pet reptiles.4 In addition, the prevalence of viruses in pet and diarrhea. The group experienced a mortality rate of
reptiles and possibly also wild reptiles appears to undergo 82%. An adenovirus was detected by polymerase chain
a fluctuating pattern in many cases, so that study over reaction (PCR) as well as by electron microscopy in cells
multiple years or decades may be necessary in order to of affected tissues. The virus was determined to belong in
understand the natural dynamics of viral infection in these the genus Siadenovirus. Siadenoviruses had previously been
animals.5 This may be in part due to functions of reptile described in birds but are hypothesized to have evolved
immunity as well as, in some cases, the long time span in amphibians.10 The same virus was later also found in
that may occur between infection and the development of impressed tortoises (Manouria impressa) and a Burmese star
disease. tortoise (Geochelone platynota) with systemic disease.11
A wide range of viruses has been described in reptiles Other adenoviruses found in chelonians have differed
(Table 39.2). This chapter focuses on select viruses that genetically from previously described genera. These viruses
have been recently described or for which there is evidence have been described in a wide range of terrestrial and
that their epidemiology has changed in recent years. Rana- aquatic species in the United States and Europe. They have
viruses are important emerging pathogens in chelonians been found in both diseased and clinically healthy animals.
267
TABLE
39.1 Arboviruses Described in Reptiles
Associated
Invertebrate Geographic Disease in Zoonotic
Virus Family Virus Name Vectors Reptilian Hosts Distribution* Reptiles Potential Ref.
Bunyaviridae Orthobunyvirus: Cache Mosquitoes, Texas soft-shelled turtle (Apalone North None Yes 55, 56
Valley virus, Tensaw virus, Anopheles spp. [Trionyx] spinifera emoryi), Skink America,
Kowanyama virus (Cryptoblepharus [Ablepharus boutonii] Australia
virgatus)
Nairovirus: Crimean-Congo Ticks (Hyalomma Horsfield’s tortoise (Testudo horsfieldii) Middle East None Yes 57
hemorrhagic fever virus aegyptium)
Togaviridae Eastern equine encephalitis Mosquitoes Various squamates, most often in various North and None Yes 58–60
virus (EEEV), Venezuelan snake spp.; chelonians, crocodilians South
equine encephalitis virus America
(VEEV), western equine
encephalitis virus (WEEV)
268 SE C T I O N 8 Emerging and Changing Infectious Diseases
Flaviviridae Japanese encephalitis virus, Mosquitoes Crocodilians, squamates, chelonians; WNV Asia, North Neurologic and Yes 59–62
St. Louis encephalitis especially in crocodilians and South skin lesions in
virus, Powasan virus, West America crocodilians
Nile virus (WNV), Zika virus
Rhabdoviridae Vesicular stomatitis virus, Mosquitoes (e.g., Redbelly water snake (Nerodia [Natrix] Australia, None reported Yes 55, 56,
Charlesville virus, Almpiwar Aedes aegypti), erythrogaster), Australian house gecko North and 63, 64
virus, Marco virus, Timbo sandflies (Gehyra australis), teiid lizards: giant South
virus, Chaco virus, Sena (Phlebotomus ameiva (Ameiva ameiva ameiva), Striped America
Madureira virus spp.), midges forest whiptail (Kentropyx calcarata),
(Forcipomyia spp., Caiman lizard (Dracaena guianensis),
Culicoides spp.) skink (Cryptoblepharus virgatus)
Iridoviridae Hemocytiviruses Unknown Various squamate and chelonian spp.; Africa, Asia, Anemia, Unlikely 65–69
sequence data available from: Europe, systemic
Bearded dragon (Pogona vitticeps), North disease
peninsula ribbon snake (Thamnophis America,
sauritus sackenii), Iberian mountain
lizard (Iberolacerta [Lacerta] monticola)
*Geographic areas in which detection in reptiles has been reported. The geographic range of individual viruses may be greater.
This table includes descriptions of detection of viruses by various methods as well as serologic evidence of previous infection (without proof of viremia).
CHAPTER 39 Emerging Reptile Viruses 269
TABLE
39.2 Virus Families and Genera Described in Orders of Reptiles
*There is some evidence of infection with these viruses in this group of reptiles.
The viruses detected in these cases have been preliminarily switched hosts into chelonian species (the siadenovirus and
named “testadenoviruses,” based on the hypothesis that atadenovirus) appear to have been associated with more
they have coevolved in chelonians.6 In another case in a severe pathologies.
spur-thighed tortoise (Testudo graeca) with stomatitis and
esophagitis, a virus in the genus Atadenovirus was detected Herpesviridae
by PCR and sequencing.12 There are therefore a multitude
of different adenoviruses belonging to at least three differ- Herpesviruses are large, enveloped dsDNA viruses that
ent genera that have been discovered in chelonians within are known to cause latent infections in hosts surviving
the past decade. The viruses hypothesized to have recently acute infection. Herpesviruses have long been described
270 SE C T I O N 8 Emerging and Changing Infectious Diseases
as pathogens in a wide range of reptilian hosts, especially have also indicated shifts in prevalence of specific viruses
chelonians. Detections in various orders of reptiles have in some pet chelonians.
increased in recent years. In many cases, virus detection There are four known genetically distinct herpesviruses
is based on histologic detection of intranuclear inclusion that can infect tortoises, named testudinid herpesvirus 1
bodies in infected cells, followed by electron microscopy through 4 (TeHV1-4). TeHV1, 2, and 3 have all been asso-
and on detection of viral DNA using a panherpesviral ciated with severe stomatitis and glossitis as well as rhinitis,
PCR targeting a portion of the DNA-dependent DNA conjunctivitis, and hepatitis. TeHV4 was originally detected
polymerase.13 in a clinically healthy tortoise during quarantine screening.25
TeHV3 has been associated with higher morbidity and mor-
Crocodilian Herpesviruses tality rates than TeHV1.26 Recent studies using molecular
techniques to investigate genomic differences between
Recent studies in Australia in farmed saltwater crocodiles TeHV3 strains as well as transmission studies have provided
(Crocodylus porosus) have led to the description of three evidence that there may also be differences in pathogenicity
different crocodilian herpesviruses.14 Infections have been between different isolates.24,27 In a study in Europe almost
associated with three disease syndromes: conjunctivitis and/ 20 years ago in which samples from tortoises kept as pets
or pharyngitis (CP), lymphoid proliferation and nonsup- were tested for the presence of herpesviruses, only TeHV1
purative encephalitis (SLPE), and multifocal lymphohis- and TeHV3 were detected, with TeHV3 making up greater
tiocytic infiltration of the dermis (LNS).5 CP was found than 80% of the detected herpesviruses.28 In a recent study
primarily in hatchlings, SLPE in juveniles and growers, and screening samples from over 1000 chelonians in Europe
LNS in harvest-sized animals. The causative nature of the for herpesviruses and other pathogens, approximately half
herpesviruses for each disease syndrome has not been fully of the herpesviruses detected were categorized as TeHV1,
established, although herpesviruses were found significantly indicating that the prevalence of this virus in pet tortoises
more often in diseased crocodiles than in controls.5 in Europe has increased over the past years, possibly due to
changes in the pet trade.3 It might also reflect a change in
Squamate Herpesviruses the popularity of specific tortoise species in the pet trade.
During that same study, a TeHV4 was detected for the first
Herpesviruses have been described periodically in various time in Europe in an African tortoise species, the leopard
squamate species. Genetically, little information is available tortoise (Stigmochelys pardalis).29
on these viruses, but what is available indicates that they are Detections of herpesviruses in other families of chelonians
diverse and not closely related to chelonian or crocodilian has also increased rapidly in recent years.30–36 In the family
herpesviruses. In lizards, herpesviruses have been associated Emydidae, herpesviruses have been detected in a freshwa-
with stomatitis,15,16 papillomas,17 hepatitis, and enteritis.18 ter turtle (Pseudemys concinna concinna), a northern map
In a disease outbreak associated with a herpesvirus infection turtle (Graptemys geographica), wild bog turtles (Glyptemys
in captive adult horned vipers (Vipera ammodytes ammodytes) muhlenbergii), wood turtles (G. insculpta), spotted turtles
in Europe, animals developed widespread hemorrhage, (Clemmy guttata), and box turtles (Terrapene sp.). Clinical
coelomic and pericardial effusion, and hepatitis. All of the signs associated with infection in these animals have ranged
horned vipers in the collection died, whereas common from detections in clinically healthy animals to stomatitis,
European vipers (Vipera berus) housed in the same facility papillomatous skin lesions, rhinitis, and sudden death.
remained unaffected.19 Histology has demonstrated hepatic lipidosis, pneumonia,
and both hepatocellular and splenic necrosis. Intranuclear
Chelonian Herpesviruses inclusions have been found in cells in the liver, lung, and
spleen.31 Infected animals have included both captive and
Herpesviruses have long been described as important patho- wild turtles.
gens in various chelonian species, mostly in sea turtles, In the order Pleurodira, herpesviruses have been described
where they are considered the cause of fibropapillomatosis in a captive Krefft’s river turtle (Emydura macquarii krefftii)
(see Chapter 57), as well as grey patch disease,20 lung, (family Chelidae) in Australia with ulcerative lesions of the
eye, and trachea disease,21 loggerhead genital-respiratory skin and shell associated with orthokeratotic hyperkeratosis
herpesvirus-associated disease, and loggerhead orocutaneous with intranuclear inclusions in keratinocytes.35 In the family
herpesvirus-associated disease,22 and in tortoises, in which Pelomedusidae, a herpesvirus was detected in West African
they have mostly been associated with stomatitis, rhinitis, and mud turtles (Pelusios castaneus) that were imported into
conjunctivitis. The chelonid fibropapillomatosis–associated Europe from Africa and were clinically healthy.36
herpesvirus, officially known as Chelonid alpha herpesvirus
5, has been placed in a separate genus, Scutavirus, in the Picornaviridae in Tortoises
subfamily Alphaherpesvirinae.23 All reported herpesviruses of
chelonians appear to cluster in this genus.24 Recent studies Picorna-like viruses have been known to occur in tortoises
on herpesviruses in a wide range of chelonian species have in Europe since the mid-1990s. They were originally iso-
led to the description of numerous new virus types and lated in cell culture and, proving difficult to characterize,
CHAPTER 39 Emerging Reptile Viruses 271
were sometimes called “virus x.” Viruses in this category depression, and loss of body condition.46 Following detec-
have been sequenced and shown to represent a new genus tion of a barnivirus-like virus in affected animals, a real-
in the family Picornaviridae, with the name Torchivirus.37,38 time PCR was developed and used to detect virus in oral
This group of viruses have been detected mostly in Testudo secretions. Virus was detected significantly more often in
spp. but also in several African tortoise species. They have diseased than in healthy animals, although virus was also
been associated with softening of the plastron in juvenile detected in some of the lizards that were considered healthy
tortoises and with rhinitis, stomatitis, and ascites in adult at the time of sampling.46
tortoises. They have also been isolated from clinically healthy
animals.28,39 Transmission studies with T. hermanni and T. Arenaviridae in Snakes
graeca showed that the kidneys were most severely affected.40
Serologic testing has shown that some wild-caught tortoises Inclusion body disease (IBD) was originally described in the
in Europe and Africa have antibodies against these viruses. early 1980s and 1990s47 and was defined by the presence of
Epidemiology of these viruses in Europe appears to undergo characteristic eosinophilic to amphophilic intracytoplasmic
some fluctuation over time. A recent study showed that inclusions in neurons and in epithelial cells of a wide range
torchiviruses were detected by PCR using mostly oral swabs of tissues.48 The inclusions are made up of a specific protein
as samples in 2.2% of over 1000 chelonians tested over a known as inclusion body disease protein (IBDP).48 The
11 2-year period in a commercial laboratory in Europe.3 disease affects various species of boas and pythons and is
Another picornavirus with the suggested genus name associated with a wide variety of clinical signs. Central
“Rafivirus” was described from Sulawesi tortoises that were nervous system (CNS) signs are most often described, but
also infected with an adenovirus.41 Animals died with severe animals may develop anorexia, pneumonia, various skin
systemic disease,9 but the role of the picornavirus in the lesions, mouth rot, and other problems. However, a large
disease is not known. number of snakes (especially boa constrictors) may remain
clinically healthy despite the presence of inclusions and/or
Nidovirales virus. In 2012, NGS showed the probable cause of IBD to
be newly discovered viruses in the family Arenaviridae in
Coronaviruses belong to the order Nidovirales, and viruses the new genus Reptarenavirus.49–51 These viruses are closely
in this family have been among the most prominent emerg- associated with the disease and can cause the development
ing viruses in a wide variety of mammalian species in recent of similar inclusions in cell culture. Antibodies against
years, causing various respiratory disease syndromes includ- reptarenaviruses isolated in cell culture bind to typical
ing severe acute respiratory syndrome (SARS) and Middle inclusions in the cells of affected animals.49,50 IBDP is
East respiratory syndrome (MERS). NGS of samples from believed to be accumulated viral nucleocapsid protein.49
reptiles displaying signs of respiratory disease has led to Additional studies on snakes infected with arenaviruses
the description of viruses in the order Nidovirales, family have demonstrated a huge amount of genetic diversity in
Coronaviridae, subfamily Torovirinae in various python these viruses, with multiple species and possibly different
species and in boas.42–45 A new genus name, “Barnivirus”, genera detected.4,52 In many cases, snakes were infected
has been suggested for these viruses.42 Infections have been with multiple viruses, and the two genomic segments
reported most commonly in ball pythons (Python regius) of the reptarenaviruses, L and S, appeared to reassort
but also in Indian pythons (P. molurus), Burmese pythons readily.4 It has been hypothesized that snake importation
(P. bivittatus), green tree pythons (Morelia viridis), carpet and husbandry practices may have created this diversity
pythons (M. spilota), and boa constrictors (Boa constrictor). among reptarenaviruses, with capture of wild snakes,
A genetically related but distinct virus has been described importation into various countries, mixing of animals
in shingleback lizards (“bobtails,” Tiliqua rugosa) in Aus- for breeding purposes, and overcrowding all being part
tralia.46 In ball pythons, the “Barnivirus” was associated of an “anthropogenic disruption of pathogen ecology.”4 It
with a proliferative interstitial pneumonia. In some cases, has been hypothesized that coinfection or superinfection
tracheitis, stomatitis, esophagitis, and/or rhinitis were also with multiple reptarenaviruses could play a role in disease
associated with infection. In individual cases, lesions were development.52
also observed in other parts of the body and included
encephalitis, acute nephritis, salpingitis, hepatic lipidosis, Other Newly Described Viruses of Reptiles
keratitis, and colitis. The greatest viral load was detected in
the lungs of affected snakes.42 A similar disease syndrome There are many other examples of recently described viruses
of unknown etiology was reported to have been observed in reptiles, with new reports coming out regularly. Recent
in ball pythons since the 1990s.42 developments include a description of the new family
The shingleback lizards, in which a related virus was Sunviridae for Sunshinevirus in the Mononegavirales. Sun-
detected, were wild caught in western Australia and found shinevirus is associated with CNS and respiratory disease
to have a disease syndrome called “bobtail flu,” with clinical in pythons, mostly in Australia.53 Bornaviruses have also
signs including mucopurulent discharge from the eyes and recently been detected in snakes in several cases,54 and there
nose, lethargy, lack of appetite, pale mucous membranes, is some indication that infections may be associated with
272 SE C T I O N 8 Emerging and Changing Infectious Diseases
neurologic disease (T. Hyndman, personal communication, 18. Hughes-Hanks JM, Schommer SK, Mitchell WJ, et al: Hepatitis
March 30, 2016). and enteritis caused by a novel herpesvirus in two monitor lizards,
J Vet Diagn Invest 22:295–299, 2010.
19. Catoi C, Gal AF, Taulescu MA, et al: Lethal herpesvirosis in 16
References captive horned vipers (Vipera ammodytes ammodytes): pathological
and ultrastructural findings, J Comp Pathol 150:341–344, 2014.
1. Duffus ALJ, Waltzek TB, Stöhr AC, et al: Distribution and 20. Rebel G, Rywlin A, Haines H: A herpesvirus agent associated
host range of ranaviruses. In Gray MJ, Chinchar VG, editors: with skin lesions of green sea turtles in aquaculture, Am J Vet Res
36:1221–1224, 1975.
Ranaviruses: lethal pathogens of ectothermic vertebrates, 2015,
21. Jacobson ER, Gaskin JM, Roelke M, et al: Conjunctivitis,
SpringerOpen, pp 9–58. tracheitis, and pneumonia associated with herpesvirus infection
2. Stöhr AC, Blahak B, Heckers KO, et al: Ranavirus infections in green sea turtles, J Am Vet Med Assoc 189:1020–1023, 1986.
associated with skin lesions in lizards, Vet Res 44:84, 2013. 22. Stacy BA, Wellehan JFX, Foley AM, et al: Two herpesviruses
3. Kolesnik E, Obiegala A, Marschang RE: Detection of Mycoplasma associated with disease in wild Atlantic loggerhead sea turtles
spp., herpesviruses, topiviruses, and ferlaviruses in diagnostic (Caretta caretta), Vet Microbiol 126:63–73, 2008.
samples from chelonians in Europe, J Vet Diagn Invest 2017. 23. Davison A, McGeoch D. Create genus Scutavirus (type species:
4. Stenglein MD, Jacobson ER, Chang L-W, et al: Widespread the currently unassigned species Chelonid herpesvirus 5) in
recombination, reassortment, and transmission of unbalanced subfamily Alphaherpesvirinae, family Herpesviridae, 2010.
compound viral genotypes in natural arenavirus infections, PLoS Retrieved March 9, 2017 from https://talk.ictvonline.org/ICTV/
Pathog 11(5):e1004900, 2015. proposals/2010.016a-eV.A.v2.Scutavirus.pdf.
5. Shilton CM, Jerrett IV, Davis S, et al: Diagnostic investigation of 24. Gandar F, Wilkie GS, Gatherer D, et al: The genome of a tortoise
new disease syndromes in farmed Australian saltwater crocodiles herpesvirus (testudinid herpesvirus 3) has a novel structure and
(Crocodylus porosus) reveals associations with herpesvirus infec- contains a large region that is not required for replication in vitro
tion, J Vet Diagn Invest 28:279–290, 2016. or virulence in vivo, J Virol 2015; pii: JVI.01794-15.
6. Doszpoly A, Wellehan JF, Jr, Childress AL, et al: Partial charac- 25. Bicknese EJ, Childress AL, Wellehan JF, Jr: A novel herpesvirus
terization of a new adenovirus lineage discovered in testudinoid of the proposed genus Chelonivirus from an asymptomatic bow-
turtles, Infect Genet Evol 17:106–112, 2013. sprit tortoise (Chersina angulata), J Zoo Wildl Med 41:353–358,
7. Harrach B: Reptile adenoviruses in cattle? Acta Vet Acad Sci Hung 2010.
48:485–490, 2000. 26. Marschang RE: Viruses infecting reptiles, Viruses 3:2087–2126,
8. Ball I, Ofner S, Funk RS, et al: Prevalence of neutralising antibod- 2011.
ies against adenoviruses in lizards and snakes, Vet J 202:176–181, 27. Origgi FC, Tecilla M, Pilo P, et al: A genomic approach to
2014. unravel host-pathogen interaction in chelonians: the example of
9. Rivera S, Wellehan JF, Jr, McManamon R, et al: Systemic adeno- testudinid herpesvirus 3, PLoS ONE 10:e0134897, 2015.
virus infection in Sulawesi tortoises (Indotestudo forsteni) caused 28. Marschang RE: Isolierung und Charakterisierung von Irido-,
by a novel siadenovirus, J Vet Diagn Invest 21:415–426, 2009. Herpes- und Reoviren aus Landschildkröten sowie Beschreibung
10. Davison AJ, Benkö M, Harrach B: Genetic content and evolu- eines nicht charakterisierten zytopathogenen Agens. Vet Med
tion of adenoviruses, J Gen Virol 84:2895–2908, 2003. Diss, Justus-Liebig-Universität Giessen, Germany, 2001.
11. Schumacher VL, Innis CJ, Garner MM, et al: Sulawesi tortoise 29. Kolesnik E, Mittenzwei F, Marschang RE: Detection of tes-
adenovirus-1 in two impressed tortoises (Manouria impressa) and tudinid herpesvirus type 4 in a leopard tortoise (Stigmochelys
a Burmese star tortoise (Geochelone platynota), J Zoo Wildl Med pardalis), Tierärztl Praxi Ausg K Kleintiere Heimtiere 44:283–286,
43:501–510, 2012. 2016.
12. Garcia-Morante B, Pénzes JJ, Costa T, et al: Hyperplastic sto- 30. Jungwirth N, Bodewes R, Osterhaus AD, et al: First report of a
matitis and esophagitis in a tortoise (Testudo graeca) associated new alphaherpesvirus in a freshwater turtle (Pseudemys concinna
with an adenovirus infection, J Vet Diagn Invest 28:579–583, concinna) kept in Germany, Vet Microbiol 170:403–407, 2014.
2016. 31. Ossiboff RJ, Newton AL, Seimon TA, et al: Emydid herpesvirus
13. Van Devanter DR, Warrener P, Bennett L, et al: Detection and 1 infection in northern map turtles (Graptemys geographica) and
analysis of diverse herpesviral species by consensus primer PCR, painted turtles (Chrysemys picta), J Vet Diagn Invest 27:392–395,
J Clin Microbiol 34:1666–1671, 1996. 2015.
14. Hyndman TH, Shilton CM, Wellehan JF, Jr, et al: Molecular 32. Ossiboff RJ, Raphael BL, Ammazzalorso AD, et al: Three novel
identification of three novel herpesviruses found in Australian herpesviruses of endangered Clemmys and Glyptemys turtles, PLoS
farmed saltwater crocodiles (Crocodylus porosus) and Australian ONE 10:e0122901, 2015.
captive freshwater crocodiles (Crocodylus johnstoni), Vet Microbiol 33. Sim RR, Norton TM, Bronson E, et al: Identification of a novel
181:183–189, 2015. herpesvirus in captive Eastern box turtles (Terrapene carolina
15. Wellehan JF, Johnson AJ, Latimer KS, et al: Varanid herpesvirus carolina), Vet Microbiol 175:218–223, 2015.
1: a novel herpesvirus associated with proliferative stomatitis in 34. Yonkers SB, Schneider R, Reavill DR, et al: Coinfection with a
green tree monitors (Varanus prasinus), Vet Microbiol 105:83–92, novel fibropapilloma-associated herpesvirus and a novel Spirorchis
2005. sp. In an eastern box turtle (Terrapene carolina) in Florida, J Vet
16. Wellehan JF, Nichols DK, Li LL, et al: Three novel herpesviruses Diagn Invest 27:408–413, 2015.
associated with stomatitis in Sudan plated lizards (Gerrhosaurus 35. Cowan ML, Raidal SR, Peters A: Herpesvirus in a captive Aus-
major) and a black-lined plated lizard (Gerrhosaurus nigrolinea- tralian Krefft’s river turtle (Emydura macquarii krefftii), Aust Vet
tus), J Zoo Wildl Med 35:50–54, 2004. J 93:46–49, 2015.
17. Literak I, Robesova B, Majlathova V, et al: Herpesvirus-associated 36. Marschang RE, Heckers KO, Heynol V, et al: [Herpesvirus
papillomatosis in a green lizard, J Wildl Dis 46:257–261, 2010. detection in clinically healthy West African mud turtles (Pelusios
CHAPTER 39 Emerging Reptile Viruses 273
castaneus)], Tierarztl Prax Ausg K Kleintiere Heimtiere 43:166–169, 53. Hyndman TH, Marschang RE, Wellehan JFX, et al: Isolation and
2015. molecular identification of Sunshine virus, a novel paramyxovirus
37. Farkas SL, Ihász K, Fehér E, et al: Sequencing and phylogenetic found in Australian snakes, Infect Genet Evol 12:1436–1446,
analysis identifies candidate members of a new picornavirus 2012.
genus in terrestrial tortoise species, Arch Virol 160:811–816, 54. Stenglein MD, Leavitt EB, Abramovitch MA, et al: Genome
2015. sequence of a bornavirus recovered from an African garter snake
38. Zell R, Delwart E, Gorbalenya AE, et al: Create 1 new species (Elapsoidea loveridgei), Genome Announc 2:e00779-14, 2014.
(Torchivirus A) in a new genus (Torchivirus), 2016. Retrieved 55. Hoff G, Trainer O: Arboviruses in reptiles: isolation of a bun-
Feb. 6, 2018 from: https://data.ictvonline.org/proposals/2016 yamwera group virus from a naturally infected turtle, J Herpetol
.018a-dS.A.v1.Torchivirus.pdf. 7:55–62, 1973.
39. Heuser W, Pendl H, Knowles NJ, et al: Soft plastron, soft carapace 56. Doherty RL: Arboviruses of Australia, Aust Vet J 48:172–180,
with skeletal abnormality in juvenile tortoises. Histopathology 1972.
and isolation of a novel picornavirus from Testudo graeca and 57. Široký P, Bělohlávek T, Papoušek I, et al: Hidden threat of
Geochelone elegans, Tierarztl Prax Ausg K Kleintiere Heimtiere tortoise ticks: high prevalence of Crimean-Congo haemorrhagic
42:310–320, 2014. fever virus in ticks Hyalomma aegyptium in the Middle East,
40. Paries S: The role of virus “x” (tortoise picornavirus nsp.) in Parasit Vectors 7:101, 2014.
kidney disease and shell weakness syndrome in various European 58. Jacobson ER: Viruses and viral diseases of reptiles. In Jacobson
tortoise species determined by experimental infection, in Proceed- ER, editor: Infectious diseases and pathology of reptiles, Boca Raton,
ings of the 2nd International Conference of Avian Herpetological FL, 2007, CRC Press, Taylor and Francis Group, pp 395–460.
and Exotic Mammal Medicine 2015; 351. 59. Kuno G: Persistence of arboviruses and antiviral antibodies
41. Ng TF, Wellehan JF, Coleman JK, et al: A tortoise-infecting in vertebrate hosts: its occurrence and impacts, Rev Med Virol
picornavirus expands the host range of the family Picornaviridae, 11:165–190, 2001.
Arch Virol 160:1319–1323, 2015. 60. Marschang RE: Virology. In Divers SJ, Stahl S, editors: Mader’s
42. Stenglein MD, Jacobson ER, Wozniak EJ, et al: Ball python reptile and amphibian medicine and surgery, ed 3. In press, Elsevier.
nidovirus: a candidate etiologic agent for severe respiratory 61. Marka A, Diamantidis A, Papa A, et al: West Nile Virus State of
disease in Python regius, MBio 5(5):e01484-14, 2014. the Art Report of MALWEST Project, Int J Environ Res Public
43. Bodewes R, Lempp C, Schürch AC, et al: Novel divergent nido- Health 10:6534–6610, 2013.
virus in a python with pneumonia, J Gen Virol 95:2480–2485, 62. Bueno MG, Martinez N, Abdalla L, et al: Animals in the Zika
2014. virus life cycle: what to expect from megadiverse Latin American
44. Uccellini L, Ossiboff RJ, de Matos RE, et al: Identification of a countries, PLoS Negl Trop Dis 10(12):e0005073, 2016.
novel nidovirus in an outbreak of fatal respiratory disease in ball 63. Doherty RL, Carley JG, Standfast HA, et al: Isolation of arbovi-
pythons (Python regius), Virol J 11:144, 2014. ruses from mosquitoes, biting midges, sandflies and vertebrates
45. Marschang RE, Kolesnik E: Detection of nidoviruses in live collected in Queensland, 1969 and 1970, Trans R Soc Trop Med
pythons and boas, Tierärztl Praxis Ausg K Kleintiere Heimtiere Hyg 67:536–543, 1973.
44:22–26, 2016. 64. Wellehan JFX, Pessier AP, Archer LL, et al: Initial sequence char-
46. O’Dea MA, Jackson B, Jackson C, et al: Discovery and partial acterization of the rhabdoviruses of squamate reptiles, including
genomic characterisation of a novel nidovirus associated with a novel rhabdovirus from a caiman lizard (Dracaena guianensis),
respiratory disease in wild shingleback lizards (Tiliqua rugosa), Vet Microbiol 158:274–279, 2012.
PLoS ONE 11:e0165209, 2016. 65. Alves de Matos AP, Paperna I, Crespo E: Experimental infec-
47. Schumacher J, Jacobson ER, Homer BL, et al: Inclusion body tion of lacertids with lizard erythrocytic viruses, Intervirology
disease in boid snakes, J Zoo Wildl Med 25:511–524, 1994. 45:150–159, 2002.
48. Chang LW, Jacobson ER: Inclusion body disease, a worldwide 66. Alves de Matos AP, Caeiro MF, Papp T, et al: New viruses from
infectious disease of boid snakes: a review, J Exot Pet Med Lacerta monticola (Serra da Estrela, Portugal): further evidence
19:216–225, 2010. for a new group of nucleo-cytoplasmic large deoxyriboviruses
49. Stenglein MD, Sanders C, Kistler AL, et al: Identification, (NCLDVs), Microsc Micoanal 17:101–108, 2011.
characterization, and in vitro culture of highly divergent arena- 67. Eid R, Radad K, Al-Shraim M: Iridoviral infection consistent
viruses from boa constrictors and annulated tree boas: candidate with lizard erythrocytic virus in Chamaelo calyptratus, Wien
etiological agents for snake inclusion body disease, MBio 3(4): Tierärztl Monatsschr 98:82–86, 2011.
e00180-12, 2012. 68. Grosset C, Wellehan JF, Jr, Owens SD, et al: Intraerythrocytic
50. Hetzel U, Sironen T, Laurinmäki P, et al: Isolation, identifica- iridovirus in central bearded dragons (Pogona vitticeps), J Vet
tion, and characterization of novel arenaviruses, the etiological Diagn Invest 26:354–364, 2014.
agents of boid inclusion body disease, J Virol 87:10918–10935, 69. Wellehan JF, Jr, Strik NI, Stacy BA, et al: Characterization of
2013. an erythrocytic virus in the family Iridoviridae from a penin-
51. Bodewes R, Kik M, Raj VS, et al: Detection of novel divergent sula ribbon snake (Thamnophis sauritus sackenii), Vet Microbiol
arenaviruses in boid snakes with inclusion body disease in the 131:115–122, 2008.
Netherlands, J Gen Virol 94:1206–1210, 2013.
52. Hepojoki J, Salmenperä P, Sironen T, et al: Arenavirus coinfec-
tions are common in snakes with boid inclusion body disease, J
Virol 89:8657–8660, 2015.
40
Emerging Diseases in Bats
JONATHAN H. EPSTEIN
274
CHAPTER 40 Emerging Diseases in Bats 275
Filoviruses (Ebola and Marburg Viruses) Ebola viruses.51 As with CoV and henipaviruses, filoviruses
appear to be geographically widespread in bat hosts in
Ebola virus was first discovered in 1976; since then there both Africa and Asia. Although some Ebola viruses and
have been more than 26 outbreaks of Ebola virus disease.41 Marburg virus have been associated with high mortality
Over the past 40 years, the natural reservoir for Ebola virus rates in people, Ebola Reston virus illustrates how genetic
has remained a mystery. Although some of the outbreaks diversity within a viral group can influence pathogenic-
were epidemiologically linked to contact with wild animals, ity in humans or domestic animals. Until there is an a
few had evidence directly linking cases to contact with bats.8 priori method for determining pathogenicity from genetics,
Human infections in Central Africa have been associated filovirus surveillance and ecologic research in bats and other
with such contact and with the consumption of infected wildlife—including work done at the International Centre
animals such as gorilla (Gorilla gorilla), chimpanzee (Pan for Medical Research, Franceville (CIRMF)42,47; the US
troglodytes), or duiker (Cephalophus spp.) carcasses.42 In Centers for Disease Control44,45,52; and the US Agency for
December 2013, an outbreak of Ebola Zaire virus, of unprec- International Development (USAID) under its Emerging
edented magnitude in West Africa, began in Guéckedou, Pandemic Threats: PREDICT program53—will help to
Guinea, following a single introduction from an unknown provide a better understanding of filovirus host ecology and
animal reservoir (hypothesized to be a bat) into the human viral genomics and inform strategies to reduce the risk of
population.43 Importantly, human social dynamics, rather Ebola virus disease and outbreaks of Marburg virus disease
than repeated introductions from an animal reservoir, (see also Chapter 19).
were responsible for the rapid and uncontrolled spread
of Ebola virus disease through Guinea, Sierra Leone, and
Liberia, underscoring the importance of human-wildlife Coronaviruses (Severe Acute
interaction in spillover and the triggering of epidemics and Respiratory Syndrome and Middle
pandemics. East Respiratory Syndrome)
However, over the past decade there has been a growing
body of evidence suggesting that multiple bat species CoVs comprise a large viral family known to infect a wide
carry Ebola viruses, whereas Marburg virus appears to variety of animals, including humans. Prior to the emer-
be primarily carried by the Egyptian fruit bat (Rousettus gence of SARS and MERS, only four CoVs were known
aegyptiacus), a common frugivorous bat found throughout to infect humans.54 The SARS pandemic of 2002–2003
the African continent and in the Middle East.44 Marburg infected more than 8000 people in 27 countries and had a
virus infection occurs seasonally in R. aegyptiacus, with peak case fatality rate of ~9%.55,56 MERS-CoV (as of June 2017)
infection rates occurring during the birthing season.45 As has infected more than 2000 people in 27 countries and had
with henipaviruses, experimental infections with Marburg a 35% case fatality rate.57 These two epidemics solidified
virus in R. aegyptiacus suggest that there is minimal pathol- CoVs as a viral family of concern for human health. SARS-
ogy and no visible signs of disease in these bats when CoV emerged from bats through the live animal markets
infected and that they may shed virus for up to 19 days of southern China in 2003.55 The close caging of various
postinoculation.46 Ebola virus Zaire has been detected in mammalian species, including bats, and the general lack of
several different bat species in Central Africa, including effective biosecurity practices in handling and butchering
the hammer-headed fruit bat (Hypsignathus monstrosus), animals in live animal markets facilitated the infection of
Franquet’s epauletted fruit bat (Epomops franquetti), and multiple species, including civets (Paguma larvata), raccoon
the little collared fruit bat (Myonycteris torquata).47 Ebola dogs (Nyctereutes procyonoides), and ferret badgers (Melogale
virus has not yet been isolated from bats; however, viral spp.), all of which were initially suspected as being the
RNA and antibodies have been detected in several species. primary source of the virus in early investigations.58 Initially,
Ebola Reston virus, a species causing disease in macaques civets were implicated as the source of SARS-CoV, and
but not humans or pigs, was detected in the common markets and farms were depopulated of civets as a control
bent-wing bat (Miniopterus schreibersi), a common insec- measure. Importantly, farmed civets outside the marketplace
tivorous bat, in the Philippines.48 Antibodies reactive to did not have evidence of SARS-CoV infection, suggesting
Ebola Zaire antigen have been found in Leishenault’s fruit an alternate reservoir.59,60 The eventual discovery of SARS-
bat (Rousettus leishenaulti) in Bangladesh, and although a like CoVs in bats was an important step in understanding
filovirus has not yet been identified, these findings suggest the natural reservoir, although early bat viral isolates did
that an immunogenically related filovirus is circulating in not use the same cell entry mechanism as SARS-CoV and
these bats.49 Novel filoviruses, yet to be characterized, have therefore were not able to cause SARS in animal models. In
been found in the cave nectar bat (Eonycterus spelea) and 2012, nearly 10 years after the initial discovery of bat SARS-
R. leishenaulti in China.50 These viruses may be closely like CoVs, a CoV much more closely related to SARS-CoV
related to those causing the immune response detected in and capable of directly infecting humans was identified
the same species in Bangladesh. The NPC-1 receptor, used among Chinese horseshoe bats (Rhinolophus sinicus) in
by filoviruses for cell entry, is conserved across several bat Yunnan, China.61 Although bats are no longer legally sold
species, which further supports a broad bat species range for in live animal markets in China, there are still communities,
CHAPTER 40 Emerging Diseases in Bats 277
including some in Yunnan, that hunt and eat Rhinolophus henipaviruses may provide insight into where surveillance
bats, raising the possibility that SARS could reemerge. should be targeted based on viral diversity hot spots. Cur-
In 2012, another novel CoV was discovered in people in rently there are geographic regions such as Latin America
Saudi Arabia.62 Ultimately named MERS-CoV, its genetic where there is a disproportionately low number of zoonotic
relationship to SARS-CoV and other beta CoVs found in viruses that have been characterized in bats, relative to bat
bats in Hong Kong led early investigations to focus on bats species richness, making this a region where surveillance
as a potential reservoir. A short RNA fragment matching efforts could potentially bring a high yield.6 Ultimately
MERS-CoV was found in an Egyptian tomb bat (Taphozous the integration of host ecology with viral discovery will be
perforatus) in Saudi Arabia, although epidemiologic studies important for understanding the risk of viral spillover. NiV
have not confirmed this species as a reservoir.63 MERS- is an important reminder that simply the presence of a
related CoVs have been found in other bat species in Asia host and virus is not sufficient for zoonotic transmission to
and Africa64; however, dromedary camels are the most likely occur. A viable “interface” or mechanism of transmission is
animal source of infection for people.65 Juvenile camels shed also needed for spillover (provided that people or domestic
MERS-CoV more frequently than adults, and infection is animals are susceptible to infection). Experimental studies
associated with a mild respiratory disease.65 Nosocomial will also be vital to clarify the pathogenesis and transmis-
transmission has also been a significant risk factor for human sibility of novel viruses. Reverse engineering has made it
MERS-CoV infection.66 In 2015, an outbreak involving 81 much easier to “rescue” or recreate viruses in the laboratory
people occurred in South Korea and was linked to hospital- from sequence data. Genetically modified mice provide an
based transmission.67 important model for the study of susceptibility to infection
The discovery of SARS-like CoVs in bats fueled further and pathogenesis in human physiology.
investigation and the discovery of a large diversity of CoVs The recent deluge of new viruses found in bats globally
in bats and the hypothesis that all human CoVs originated warrants a degree of caution against overstating their threat
in bats.64,68 It is estimated that 1200–6000 CoVs are carried to human or animal health when communicating findings
by bats worldwide, some of which will also have the poten- to the public or policy makers. In the majority of instances,
tial to emerge in human or domestic animal populations.64 there is no evidence that any newly discovered virus in bats
Porcine epidemic diarrhea virus (PEDV) is an alphacoro- has infected any other animal or person, thereby making
navirus that in 2013 emerged in the United States and it simply a bat virus until proven otherwise. When newly
reemerged in Asia, causing economically significant disease identified viruses are related to known zoonoses, they are
outbreaks in domestic pigs.69 Although PEDV has not been often presented as potential threats to human or animal
directly linked to bats, it does cluster phylogenetically with health, but there is the potential to cause undue public
other alphacoronaviruses that have been found in bats.64,70 alarm when reporting these findings. Given the potential
CoV diversity and richness correlate with bat species rich- for negative and scientifically unsupported actions against
ness, but as with all categories of novel viruses, it is not bats that include extermination, messaging to the public
currently possible to determine which viruses have zoonotic should provide appropriate context where there is a lack of
potential.64 Hospital- or community-based severe acute evidence for human or animal health impact and emphasize
respiratory infection (SARI) surveillance in regions with that bats are ecologically invaluable animals. Extermination
high bat biodiversity and high-risk bat-human interfaces of bats should not be considered an effective response to
(e.g., guano mining71) should consider CoV screening as an outbreak of a bat-borne pathogen or a control measure
part of a diagnostic approach to investigating respiratory to prevent outbreaks. This approach actually enhanced
disease clusters in people or diarrheal disease in animals. the local transmission of Marburg virus among bat
Identification of novel CoVs as etiologic agents in humans populations following the extirpation of bats from a mine
or domestic animals will provide additional insights into in Uganda.72
genetic determinants of pathogenicity and their relationship There will continue to be a large research and surveil-
with bat CoVs. lance focus on bats as hosts for zoonoses. Data are mount-
ing to support bats as important reservoirs compared
Discussion with other mammals, and large-scale surveillance efforts
like PREDICT and the recently launched Global Virome
Within each of the groups of viruses discussed, it is likely that Project, a 10-year effort to identify the majority of viruses in
there are still many as yet undiscovered species, strains, and key wildlife species in emerging disease hot spots,73 will shed
genetic variants comprised by the genetic diversity of nature. more light on the total diversity of viruses in bat species
In addition, the high mutation rate of RNA viruses—and and the types of human-animal interfaces that exist in
in the case of CoVs the ability to recombine—means that different geographic and cultural contexts. Understanding
new genotypes are continuously being created. This presents specific human behaviors that promote contact with bats
a serious challenge to cataloging viral diversity, but doing and developing strategies that limit bat-human-domestic
so may ultimately pay off by allowing for the identifica- animal contact without harming bats is key to reducing
tion of genetic determinants of pathogenesis. Also, having the risk of viral spillover while also preserving bats and the
a library of sequences from all bat CoVs, filoviruses, or ecologic services they provide.
278 SE C T I O N 8 Emerging and Changing Infectious Diseases
References 22. Luby SP, Hossain MJ, Gurley ES, et al: Recurrent zoonotic trans-
mission of Nipah virus into humans, Bangladesh, 2001–2007,
1. Taylor LH, Latham SM, Woolhouse MEJ: Risk factors for human Emerg Infect Dis 15:1229–1235, 2009.
disease emergence, Philos Trans R Soc London B 356:983–989, 23. WHO: Nipah virus outbreaks in the WHO South-East Asia
2001. Region: World Health Organization Southeast Asia Office, 2015.
2. Morse SS, Mazet JAK, Woolhouse M, et al: Zoonoses 3 Pre- 24. Halpin K, Hyatt AD, Fogarty R, et al: Pteropid bats are con-
diction and prevention of the next pandemic zoonosis, Lancet firmed as the reservoir hosts of henipaviruses: a comprehensive
380:1956–1965, 2012. experimental study of virus transmission, Am J Trop Med Hyg
3. Daszak P, Cunningham AA: Anthropogenic change, biodiver- 85:946–951, 2011.
sity loss, and a new agenda for emerging diseases, J Parasitol 25. Middleton DJ, Morrissy CJ, van der Heide BM, et al: Experimen-
89(Suppl):S37–S41, 2003. tal Nipah virus infection in pteropid bats (Pteropus poliocephalus),
4. Wolfe ND, Dunavan CP, Diamond J: Origins of major human J Comp Pathol 136:266–272, 2007.
infectious diseases, Nature 447:279–283, 2007. 26. Salah Uddin Khan M, Hossain J, Gurley E, et al: Use of infrared
5. Daszak P, Cunningham AA, Hyatt AD: Anthropogenic envi- camera to understand bats’ access to date palm sap: implications
ronmental change and the emergence of infectious diseases in for preventing nipah virus transmission, Ecohealth 1–9, 2011.
wildlife, Acta Trop 78:103–116, 2001. 27. Bates PJJ, Harrison DL: Sub-Order MEGACHIROPTERA:
6. Olival KJ, Hosseini PR, Zambrana-Torrelio C, et al: Host and Family Pteropodidae: old world fruit bats. Bats of the Indian
viral traits predict zoonotic spillover from mammals, Nature subcontinent. Kent, England: Harrison Zoological Museum,
2017; advance online publication. 1997;13–15.
7. Schountz T: Immunology of bats and their viruses: challenges 28. Epstein JH, Prakash V, Smith CS, et al: Henipavirus infec-
and opportunities, Viruses-Basel 6(12):4880–4881, 2014. tion in fruit bats (Pteropus giganteus), India, Emerg Infect Dis
8. Epstein JH, Field HE: Anthropogenic epidemics: the ecology of 14:1309–1311, 2008.
bat borne viruses and our role in their emergence. In Wang LF, 29. Hsu VP, Hossain MJ, Parashar UD, et al: Nipah virus encepha-
Cowled C, editors: Bats and viruses: a new frontier of emerging litis reemergence, Bangladesh, Emerg Infect Dis 10:2082–2087,
infectious diseases, 2015, Wiley-Blackwell. 2004.
9. Teeling EC, Springer MS, Madsen O, et al: A molecular phy- 30. Yadav PD, Raut CG, Shete AM, et al: Short report: detection of
logeny for bats illuminates biogeography and the fossil record, nipah virus RNA in fruit bat (Pteropus giganteus) from India, Am
Science 307:580–584, 2005. J Trop Med Hyg 87:576–578, 2012.
10. Olival KJ, Hayman DTS: Filoviruses in bats: current knowledge 31. Anthony SJ, Epstein JH, Murray KA, et al: A strategy to estimate
and future directions, Viruses-Basel 6:1759–1788, 2014. unknown viral diversity in mammals, MBio 4:2013.
11. Hahn MB, Epstein JH, Gurley ES, et al: Roosting behaviour 32. Chowdhury S, Khan SU, Crameri G, et al: Serological evidence
and habitat selection of Pteropus giganteus reveal potential of Henipavirus exposure in cattle, goats and pigs in Bangladesh,
links to Nipah virus epidemiology, J Appl Ecol 51:376–387, Plos Negl Trop Dis 2014.
2014. 33. Field H, de Jong C, Melville D, et al: Hendra virus infection
12. Nowak RM: Nowak RM, editor: Walker’s bats of the world, ed 5, dynamics in Australian fruit bats, PLoS ONE 6:2011.
Baltimore, MD, 1994, The Johns Hopkins University Press, pp 34. Epstein JH, Field HE, Luby S, et al: Nipah virus: impact, origins,
237–239. and causes of emergence, Curr Infect Dis Rep 8:59–65, 2006.
13. Hutson AM, Mickelburgh SP, Racey PA: Microchiropteran 35. Pernet O, Schneider BS, Beaty SM, et al: Evidence for henipa-
bats: global status survey and conservation action plan, Gland, virus spillover into human populations in Africa, Nat Commun
Switzerland, and Cambridge, UK, 2001, IUCN/SSC Chiroptera 5:2014.
Specialist Group. 36. Hayman DTS, Suu-Ire R, Breed AC, et al: Evidence of Henipa-
14. Boyles JG, Cryan PM, McCracken GF, et al: Economic impor- virus infection in West African fruit bats, PLoS ONE 3:2008.
tance of bats in agriculture, Science 332:41–42, 2011. 37. Li Y, Wang J, Hickey AC, et al: Antibodies to Nipah or Nipah-
15. Fujita MS, Tuttle MD: Flying foxes (Chiroptera, Pteropodidae): like viruses in bats, China, Emerg Infect Dis 14:1974–1976,
threatened animals of key ecological and economic importance, 2008.
Conserv Biol 5:455–463, 1991. 38. Broder CC, Xu K, Nikolov DB, et al: A treatment for and
16. Mickleburgh SP, Hutson AM, Racey PA: A review of the global vaccine against the deadly Hendra and Nipah viruses, Antiviral
conservation status of bats, Oryx 36:18–34, 2002. Res 100:8–13, 2013.
17. Epstein JH, Anthony SJ: Viral discovery as a tool for pandemic 39. Broder CC, Weir DL, Reid PA: Hendra virus and Nipah virus
preparedness, OIE Sci Tech Rev 36:2017. animal vaccines, Vaccine 34:3525–3534, 2016.
18. Chua K, Bellini W, Rota P, et al: Nipah virus: a recently emergent 40. Luby SP: The pandemic potential of Nipah virus, Antiviral Res
deadly paramyxovirus, Science 288:1432–1435, 2000. 100:38–43, 2013.
19. Pulliam JRC, Epstein JH, Dushoff J, et al: Agricultural inten- 41. Outbreaks Chronology: Ebola Virus Disease: CDC, 2017.
sification, priming for persistence and the emergence of Nipah 42. Leroy EM, Rouquet P, Formenty P, et al: Multiple Ebola virus
virus: a lethal bat-borne zoonosis, J R Soc Interface 9:89–101, transmission events and rapid decline of central African wildlife,
2012. Science 303:387–390, 2004.
20. Field H, Crameri G, Kung NY-H, et al: Ecological aspects of 43. Gire SK, Goba A, Andersen KG, et al: Genomic surveillance
Hendra Virus. In Lee B, Rota PA, editors: Henipavirus: ecology, elucidates Ebola virus origin and transmission during the 2014
molecular virology, and pathogenesis, 2012, pp 11–23. outbreak, Science 345:1369–1372, 2014.
21. Rahman SA., Hassan L, Epstein JH, et al: Risk factors for Nipah 44. Towner JS, Amman BR, Sealy TK, et al: Isolation of genetically
virus infection among pteropid bats, peninsular Malaysia, Emerg diverse Marburg viruses from Egyptian fruit bats, PLoS Pathog
Infect Dis 2013. 5:2009.
CHAPTER 40 Emerging Diseases in Bats 279
45. Amman BR, Carroll SA, Reed ZD, et al: Seasonal pulses of coronavirus-like virus in palm civets at an animal market and on
Marburg virus circulation in juvenile Rousettus aegyptiacus bats farms, J Virol 79:11892–11900, 2005.
coincide with periods of increased risk of human infection, PLoS 61. Ge X-Y, Li J-L, Yang X-L, et al: Isolation and characterization of
Pathog 8:e1002877, 2012. a bat SARS-like coronavirus that uses the ACE2 receptor, Nature
46. Schuh AJ, Amman BR, Jones MEB, et al: Modelling filovirus 503:535–538, 2013.
maintenance in nature by experimental transmission of Marburg 62. Zaki AM, van Boheemen S, Bestebroer TM, et al: Isolation of a
virus between Egyptian rousette bats, Nat Commun 8:2017. novel coronavirus from a man with pneumonia in Saudi Arabia,
47. Leroy EM, Kumulungui B, Pourrut X, et al: Fruit bats as reser- N Engl J Med 367:1814–1820, 2012.
voirs of Ebola virus, Nature 438:575–576, 2005. 63. Memish ZA, Mishra N, Olival KJ, et al: Middle East Respiratory
48. Jayme SI, Field HE, de Jong C, et al: Molecular evidence of Syndrome Coronavirus in Bats, Saudi Arabia, Emerg Infect Dis
Ebola Reston virus infection in Philippine bats, Virol J 12:2015. 19:1819–1823, 2013.
49. Olival KJ, Islam A, Yu M, et al: Ebola virus antibodies in fruit 64. Anthony SJ, Johnson CK, Greig DJ, et al: Global patterns in
bats, Bangladesh, Emerg Infect Dis 19:270–273, 2013. coronavirus diversity, Virus Evol 3:vex012, 2017.
50. Xing-Lou Y, Yun-Zhi Z, Ren-Di J, et al: Genetically diverse 65. Hemida MG, Elmoslemany A, Al-Hizab F, et al: Dromedary
filoviruses in Rousettus and Eonycteris spp. Bats, China, 2009 and camels and the transmission of Middle East Respiratory Syndrome
2015, Emerg Infect Dis 23:482, 2017. Coronavirus (MERS-CoV), Transbound Emerg Dis 64:344–353,
51. Ng M, Ndungo E, Kaczmarek ME, et al: Filovirus receptor 2017.
NPC1 contributes to species-specific patterns of ebolavirus 66. Mohd HA, Memish ZA, Alfaraj SH, et al: Predictors of MERS-
susceptibility in bats, Elife 4:2015. CoV infection: a large case control study of patients presenting
52. Amman BR, Jones MEB, Sealy TK, et al: Oral shedding of with ILI at a MERS-CoV referral hospital in Saudi Arabia, Travel
Marburg virus in experimentally infected Egyptian fruit bats Med Infect Dis 14:464–470, 2016.
(Rousettus aegyptiacus), J Wildl Dis 51:113–124, 2015. 67. Oh M-d, Choe PG, Oh HS, et al: Middle East Respiratory Syn-
53. 2016 PREDICT Annual Report. Davis, CA: PREDICT Con- drome Coronavirus superspreading event involving 81 persons,
sortium, 2017. Korea 2015, J Korean Med Sci 30:1701–1705, 2015.
54. van der Hoek L: Human coronaviruses: what do they cause?, 68. Drexler JF, Corman VM, Drosten C: Ecology, evolution and
Antivir Ther 12:651–658, 2007. classification of bat coronaviruses in the aftermath of SARS,
55. Ksiazek TG, Erdman D, Goldsmith CS, et al: A novel corona- Antiviral Res 101:45–56, 2014.
virus associated with severe acute respiratory syndrome, NEJM 69. Lee C: Porcine epidemic diarrhea virus: an emerging and re-
348:1953–1966, 2003. emerging epizootic swine virus, Virol J 12:193, 2015.
56. WHO: Summary of probable SARS cases with onset of illness 70. Simas PVM, Barnabe ACD, Duraes-Carvalho R, et al: Bat
from 1 November 2002 to 31 July 2003, 2003. Coronavirus in Brazil related to Appalachian Ridge and porcine
57. WHO: Middle East respiratory syndrome coronavirus (MERS- epidemic diarrhea viruses, Emerg Infect Dis 21:729–731, 2015.
CoV), 2017. 71. Wacharapluesadee S, Sintunawa C, Kaewpom T, et al: Group C
58. Guan Y, Zheng BJ, He YQ, et al: Isolation and characteriza- Betacoronavirus in Bat Guano Fertilizer, Thailand, Emerg Infect
tion of viruses related to the SARS coronavirus from animals in Dis 19:1349–1351, 2013.
Southern China, Science 302:276–278, 2003. 72. Amman BR, Nyakarahuka L, McElroy AK, et al: Marburgvirus
59. Tu CC, Crameri G, Kong XG, et al: Antibodies to SARS coro- resurgence in Kitaka Mine bat population after extermination
navirus in civets, Emerg Infect Dis 10:2244–2248, 2004. attempts, Uganda, Emerg Infect Dis 20:1761–1764, 2014.
60. Kan B, Wang M, Jing HQ, et al: Molecular evolution analysis 73. Carroll D, Daszak P, Wolfe ND, et al: The Global Virome
and geographic investigation of severe acute respiratory syndrome Project. in review.
41
Zika Virus: A Real Threat to Wildlife?
LILIAN SILVA CATENACCI AND BIANCA NASCIMENTO DE ALCANTARA
280
CHAPTER 41 Zika Virus: A Real Threat to Wildlife? 281
TABLE Wildlife Species That Have Tested Positive for Zika Virus Antibody or Antigen by Serologic,
41.1 Molecular,and/or Viral Isolation Testing in Experimental and/or Natural Conditions
*Experimental infection; †natural infection; ‡serologic diagnostic; §molecular diagnostic; ¶virus isolation diagnostic; NA, not mentioned.
Data from Bueno MG, Martinez N, Abdalla L, et al: Animals in the Zika virus life cycle: what to expect from megadiverse Latin American countries. PLoS Negl
Trop Dis 10, 2016; Haddow AD, Schuh AJ, Yasuda CY, et al: Genetic characterization of Zika virus strains: geographic expansion of the Asian lineage. PLoS
Negl Trop Dis 6:e1477, 2012; and Vanchiere JA, Ruiz JC, Brady AG et al: Experimental Zika Virus Infection of Neotropical Primates. Am J Trop Med Hyg
98:173–177, 2017.
282 SE C T I O N 8 Emerging and Changing Infectious Diseases
Few studies have focused on the role of animals as hosts In response to the ongoing epidemic, new studies on
for ZIKV.16,24 In 1956, for the first time, Boorman and ZIKV have been carried out in model animals, mainly to
Porterfield were able to confirm the transmission of ZIKV address research and development needs for novel methods
from A. aegypti to animals, especially mice and monkeys of diagnosis, vaccination, and therapy for human popula-
during experimental studies.28 Multiple monkey species in tions. Nevertheless, research is still greatly needed to better
the Zika Forest were found to be seropositive for ZIKV, understand the risk of ZIKV for wildlife.
suggesting that they may become infected and support
viral replication.26 However, other mammals in the Zika Pathogenesis of Zika Virus
Forest (including squirrels, tree rats, giant pouched rats, and
civets) did not show serologic evidence of ZIKV infection,19 Mosquito-borne flaviviruses are thought to replicate ini-
although a subsequent study in Kenya detected ZIKV anti- tially in dendritic cells near the site of the infectious bite
bodies in small mammals including rats and shrews.29 A and then to spread quickly to lymph nodes, where they
study by Bueno and collaborators (2016) presented wildlife replicate and thereafter reach the bloodstream.26 In vitro,
species that were antibody-positive to ZIKV, both in situ ZIKV has been shown to infect fibroblasts and keratin-
and experimentally. Most primates identified as ZIKV- ocytes.26 Despite recent progress in our understanding, the
positive in the wild or in sentinel studies are Old World pathogenesis of ZIKV in vivo remains largely unknown.26
species.24 To date, there has not been solid evidence of an Similarly, ZIKV distribution in anatomic tissues, the
Asian or American sylvatic cycle of ZIKV, but such a sylvatic duration of infectious shedding, and the immune response
cycle could be widespread and still go undetected due to to primary ZIKV in humans and other animals remain
the lack of surveillance for sylvatic arboviruses.9,24 The pres- unclear.21
ence of animals seropositive for ZIKV does not necessarily Several species of immunocompromised mice29 and
mean that they are viremic or may be able to transmit the NHPs, such as rhesus, pigtail (Macaca nemestrina), and
virus to a mosquito (as a reservoir). Further studies are cynomolgus (Macaca fascicularis), were used for studies
required to properly understand the role of vertebrates in of the natural behavior and pathogenesis of ZIKV infec-
the ZIKV dynamic transmission.9,18,24 Recently ZIKV has tion.5,12,21,26 Studies also have been conducted to better
been of significant concern for conservationists, and there understand the serologic behavior and pathogenesis caused
is a lack of studies in wildlife species.1,13,30 Free-living and by the Asian lineage in neotropical primates (Callimico
semicaptive orangutans (Pongo spp.) were evaluated during goeldi, Saimiri collinsi, Saimiri boliviensis boliviensis, Saimiri
translocations from forest fragments or degraded habitat sciureus sciureus and Aotus nancymaae).32–36 These studies
in eastern Sabah (Malaysia) in 2003 and were found to evaluated the experimental infection by ZIKV, the inocula-
have anti-ZIKV antibodies.30 But how this virus could be tion routes, and the viral load in the viremic period in an
pathogenic for wildlife that already suffer anthropogenic effort to evaluate these NHPs as ZIKV study models.32–36
and natural pressure remains unknown. Is the ZIKV All assays of rhesus and cynomolgus monkeys have shown
capable of leading to NHP population declines, as has to be permissive for infection by the Asian strain of ZIKV.
occurred for howler monkey (Alouatta spp.) due to yellow Virus RNA was detected in several body fluids, and it was
fever outbreaks?9,18,24 The role of neotropical primates in the possible to detect different concentrations of this nucleic
maintenance cycle of ZIKV is still unclear.31 Could other acid in different body fluids such as plasma, urine, saliva,
vectors—such as Haemagogus sp. or Sabethes sp.—serve as cerebrospinal fluid, semen, seminal fluids, and vaginal
bridges facilitating the reverse spillover and establishment of fluids 1–10 days postinfection (DPI).5,12,25,26 Cynomolgus
an enzootic ZIKV transmission cycle in the Americas?9,24,31 monkeys presented higher viral loads of ZIKV in testis 8
Based on the huge biodiversity in the Americas, the proxim- DPI. After 2–3 weeks of infection, neutralizing antibodies
ity of wild vertebrate species to urban and rural areas, and to ZIKV were detected in most animals.5,12,26 Cynomolgus
the wide distribution of A. aegypti and other mosquito monkeys were also challenged with the African lineage, but
genera, ZIKV spillover to wild primates or another wildlife they were not permissive.5 Dudley et al. (2016) and Osuna
vertebrate is a potentially real scenario.9,18,24 et al. (2016) reinfected rhesus monkeys and did not observe
In northeastern Brazil, studies showed that 29% (7/24) viral replication, which could mean that the monkeys
of free-living and asymptomatic New World primates, Cal- may acquire immune protection after the first ZIKV
lithrix jacchus and Sapajus libidinosus, were infected with exposure.
ZIKV.31 Using real-time reverse-transcriptase polymerase According to Vanchiere et al. (2017), viremia and viruria
chain reaction (qRT-PCR), they detected the virus and were detected in two (Saimiri boliviensis boliviensis, Saimiri
showed that the ZIKV genome sequence from monkeys sciureus sciureus) of the four total animals by RT-qPCR. Peak
was 100% similar to the ZIKV circulating in humans in plasma viremia occurred between 5–10 DPI as extrapolated
South America.31 To our knowledge, no other neotropical from qPCR. Viruria was detected at 9 and 14 dpi; salivary
species were detected with ZIKV in natural conditions. ZIKV secretion was detected in three animals as late as
However, Oliveira-Filho et al. (2018) found the presence 14 dpi.
of neutralizing antibodies for ZIKV in S. libidinosus (1/49) Two of the four owl monkeys (Aotus nancymaae) had
and S. flavius (1/49) in natural conditions in Brazil. ZIKV viremia at 2 and 4 dpi by PCR. However, ZIKV was
CHAPTER 41 Zika Virus: A Real Threat to Wildlife? 283
not detected in urine, salivary secretions, or excretions from investigators, aspartate aminotransferase, alanine amino-
owl monkeys by culture or qPCR assays.36 transferase, alkaline phosphatase, and creatinine phosphatase
increased and peaked by day 3 and day 21.21,25,26 Creatine
Clinical Manifestations kinase increases may be due to viral myositis or repeated
sedation hemolysis and endocrine abnormalities.25
As occurrence of ZIKV infections in wildlife was mainly The pattern of white blood cell counts remained within
found accidentally during serosurveys searching for other normal variation ranges,25,26 although increases were found
pathogens, there is almost no information on clinical signs by Osuna et al. (2016). However, the counts returned to
in wild animals.35 In a sentinel study in Uganda in 1947, baseline in most animals within 2 days of infection.21
one primate showed mild pyrexia.12 Typically ZIKV infec- All neotropical primates infected by Vanchiere et al.
tion in humans has been associated with a self-limiting (2017) remained clinically well after inoculation. The
febrile illness often including rash, arthralgia, and conjunc- routine hematology and serum chemistry analyses were
tivitis, although most infections are asymptomatic.13,29 Wild normal at 4 dpi, with the exception of a modest increase in
mammals with ZIKV infection apparently have few clinical ALT over baseline for one animal.36
signs.24
A study by Dudley et al. (2016) in rhesus macaques Zika Virus Targets Various Organs and Causes
infected with ZIKV showed mild to moderate inappetence Pathologic Damage
resulting in mild weight loss in four animals. Two animals
also developed a very mild rash around the inoculation site at Necropsy data from rhesus and cynomolgus macaques dem-
1 DPI that persisted for 4–5 days. No other abnormal clini- onstrate that ZIKV strains may invade and replicate within
cal signs were noted. Two other studies in rhesus macaques the CNS (including cerebrum, cerebellum, brain stem, and
demonstrated that within 8–10 DPI animals displayed fever spinal cord) of macaques following subcutaneous infection
(axillary temperature 38.9°C), with peak temperatures of despite the fact that no neurologic signs are observed.21,26
40.1°C23 and 39.5°C.21 All experiments used almost the ZIKV RNA has also been detected in visceral fragments
same infectious dose of ZIKV (approximately 105 plaque- (e.g., liver, kidney, spleen, parotid glands, large intestine,
forming unit [PFU]) but from different strains, which may small intestine, cecum, bladder, testes, lymph node, heart,
explain the difference in results. and stomach) in rhesus macaques. Male genital tracts were
One of the six cynomolgus monkeys that had been chal- identified with persistent foci of ZIKV-infected cells local-
lenged with the ZIKV African lineage developed a slight ized in the testes, prostate, and seminal vesicles in the two
erythema around the injection site (Draize dermal score of 1 species studied.21,37 This observation may have negative
on study days 8 and 10 DPI). The erythema had resolved by implications for conservation programs if it is demonstrated
day 14. No clinical signs were observed in any of the other that ZIKV can be transmitted sexually.
animals during the course of the study.5 As this individual
did not present viremia of the ZIKV African lineage, the
erythema was probably not related with the virus. Collecting Biological Materials to
None of the neotropical primates infected by the Investigate Zika Virus in Wildlife
Vanchiere et al. (2017) exhibited signs of clinical disease
after ZIKV inoculation. Once collected, blood samples should be divided into two
No evidence of neurologic complications, such as aliquots. A whole-blood sample is immediately stored in
the Guillain-Barré syndrome observed in human adults, liquid nitrogen or dry ice; the other is maintained at 4°C
was found in the experiments with monkeys. However, for 3 hours until it is centrifuged for the collection of
congenital birth defects were demonstrated in one experi- serum, which is then placed in liquid nitrogen. Samples
ment.33 Ten days after inoculation with a ZIKV Asian should be sent on dry ice to the diagnostic reference
lineage strain in a pregnant pigtail macaque at 119 days of center. A minimum of 0.5 mL of whole blood is required
gestation, the fetal brain developed a periventricular lesion to perform molecular assays for the detection of virus and
and evolved asymmetrically in the occipitoparietal lobes. 0.5 mL of serum is the minimum necessary to conduct
However, in this experiment, five subcutaneous infections antibody screening. For the detection and isolation of
in the pregnant animal were initiated using 107 PFU each.33 ZIKV, whole blood should be collected in the viremic
This amount of ZIKV inoculated is higher than that used period (1–5 DPI); it may be detected by qRT-PCR up to
in the other experiments and may not represent natural 14 DPI depending on the inoculated viral load. After 7
infection. DPI, antibody testing is prioritized. Molecular detection in
other fluids such as urine, saliva, and tears and also tissues
Complete Blood Count and Blood such as CNS and placenta occurs well during the viremic
Chemistry Panels period and should be stored in liquid nitrogen and trans-
ported on dry ice. Tissues for histopathologic processing
Blood chemistry was monitored in three ZIKV infection should be stored in 10% formaldehyde and sent at room
assays.21,25,26 According to the observations made by the temperature.38
284 SE C T I O N 8 Emerging and Changing Infectious Diseases
A B
C D
• Figure 41.1 (A) Fetal necropsy in Saimiri collinsi; (B) Brain infected with Zika virus, showing lisencephaly
and dilation of the 4th cerebral ventricle; (C and D) Frontal cortical area with vasocongestion, focal
hemorrage with gliosis, satelitosis and neuroniophagia, besides of immunostaining in neurons and glial
cells. (Alcantara BN personal file). (Images courtesy Bianca Alcantara.)
7. Faria NR, Azevedo RDSDS, Kraemer MUG, et al: Zika virus in 26. Li X-F, Dong H-L, Huang X-Y, et al: Characterization of a
the Americas: Early epidemiological and genetic findings, Science 2016 clinical isolate of Zika virus in non-human primates,
352:345–349, 2016. EBioMedicine 12:170–177, 2016.
8. Mota MT, Terzian AC, Silva ML, et al: Mosquito-transmitted 27. Huang W-C, Abraham R, Shim B-S, et al: Zika virus infection
viruses - the great Brazilian challenge, Braz J Microbiol 47 (Suppl during the period of maximal brain growth causes microcephaly
1):38–50, 2016. and corticospinal neuron apoptosis in wild type mice, Sci Rep
9. Althouse BM, Vasilakis N, Sall AA, et al: Potential for Zika virus 6:34793, 2016.
to establish a sylvatic transmission cycle in the Americas, PLoS 28. Boorman JPT, Porterfield JS: A simple technique for infection of
Negl Trop Dis 10:e0005055, 2016. mosquitoes with viruses transmission of zika virus, Trans R Soc
10. Oehler E, Watrin L, Larre P, et al: Zika virus infection compli- Trop Med Hyg 50:238–242, 1956.
cated by Guillain-Barre syndrome–case report, French Polynesia, 29. Lazear HM, Govero J, Smith AM, et al: A mouse model of Zika
December 2013, Euro Surveill 19:2014. virus pathogenesis, Cell Host Microbe 19:720–730, 2016.
11. Araujo LM, Ferreira MLB, Nascimento OJ, et al: Guillain-Barré 30. Kilbourn AM, Karesh WB, Wolfe ND, et al: Health evaluation
syndrome associated with the Zika virus outbreak in Brazil, Arq of free-ranging and semi-captive orangutans (Pongo pygmaeus) in
Neuropsiquiatr 74:253–255, 2016. Sabah, Malaysia, J Wildl Dis 39:73–83, 2003.
12. Dick GWA, Kitchen SF, Haddow AJ: Zika virus. I. Isolations and 31. Favoretto S, Araujo D, Oliveira D, et al: First detection of Zika
serological specificity, Trans R Soc Trop Med Hyg 46:509–520, virus in neotropical primates in Brazil: a possible new reservoir,
1952. bioRxiv 049395, 2016.
13. Hayes EB: Zika virus outside Africa, Emerg Infect Dis 15: 32. Alcantara BN, Simith DB, Araújo MTF, et al: Standardization
1347–1350, 2009. of Zika virus antigen detection by immunohystochemistry in
14. Beckham JD, Pastula DM, Massey A, et al: Zika virus as an neotropical primate tissues. In: Proceedings; 2017.
emerging global pathogen: neurological complications of zika 33. Simith DB, Alcantara BN, Carvalho CAM, et al: Detection of
virus, JAMA Neurol 73:875–879, 2016. IgM antibodies by ELISA in the non-human primate Saimiri
15. Aman MJ, Kashanchi F: Zika virus: a new animal model for Collinsi experimentally infected by Zika virus, in proceedings.
an arbovirus, PLoS Negl Trop Dis 10:2016. Available at: http:// 2017. In: Proceedings; 2017.
www.ncbi.nlm.nih.gov/pmc/articles/PMC4858223/. (Accessed 34. Alcantara BN, Rodrigues DSG: Neotropical Callimico goeldii is
29 January 2017). susceptible to Zika virus intradermal infection. Abstract present
16. Haddow AD, Schuh AJ, Yasuda CY, et al: Genetic characteriza- in First International Conference on Zika Virus, 2017, Washing-
tion of Zika virus strains: geographic expansion of the Asian ton DC. 2017. https://www.zikaconference.com/.
lineage, PLoS Negl Trop Dis 6:e1477, 2012. 35. Oliveira-Filho EF, Oliveira RAS, Ferreira DRA, et al: Seropreva-
17. Parra B, Lizarazo J, Jiménez-Arango JA, et al: Guillain–Barré lence of selected flaviviruses in free-living and captive capuchin
Syndrome associated with Zika virus infection in Colombia, N monkeys in the state of Pernambuco, Brazil, Transbound Emerg
Engl J Med 375:1513–1523, 2016. Dis 2018. https://doi.org/10.1111/tbed.12829.
18. Vorou R: Zika virus, vectors, reservoirs, amplifying hosts, and 36. Vanchiere JA, Ruiz JC, Brady AG, et al: Experimental Zika
their potential to spread worldwide: what we know and what we virus infection of neotropical primates, Am J Trop Med Hyg
should investigate urgently, Int J Infect Dis 48:85–90, 2016. 98:173–177, 2017.
19. Haddow AJ, Williams MC, Woodall JP, et al: Twelve isolations of 37. Adams Waldorf KM, Stencel-Baerenwald JE, Kapur RP, et al:
Zika virus from Aedes (Stegomyia) africanus (Theobald) taken in Fetal brain lesions after subcutaneous inoculation of Zika virus in
and above a Uganda forest, Bull World Health Organ 31:57–69, a pregnant nonhuman primate, Nat Med 22:1256–1259, 2016.
1964. 38. CDC: CDC’s Response to Zika virus: Collection and submission
20. Paul LM, Carlin ER, Jenkins MM, et al: Dengue virus antibodies of fetal tisuues for zika virus testing. In: Proceedings: 1–3, 2016.
enhance Zika virus infection, bioRxiv 050112, 2016. 39. Zarnke RL, Calisher CH, Kerschner J: Serologic evidence of
21. Osuna CE, Lim S-Y, Deleage C, et al: Zika viral dynamics arbovirus infections in humans and wild animals in Alaska, J
and shedding in rhesus and cynomolgus macaques, Nat Med Wildl Dis 19:175–179, 1983.
22:1448–1455, 2016. 40. Clarke DH, Casals J: Techniques for hemagglutination and
22. Gyawali N, Bradbury RS, Taylor-Robinson AW: The global hemagglutination-inhibition with arthropod-borne viruses, Am
spread of Zika virus: Is public and media concern justified in J Trop Med Hyg 7:561–573, 1958.
regions currently unaffected? Infect Dis Poverty 5:37, 2016. 41. Mlakar J, Korva M, Tul N, et al: Zika virus associated with
23. Heymann DL, Hodgson A, Sall AA, et al: Zika virus and micro- microcephaly, N Engl J Med 374:951–958, 2016.
cephaly: Why is this situation a PHEIC? Lancet 387:719–721, 42. Sarno M, Aquino M, Pimentel K, et al: Progressive lesions of
2016. central nervous system in microcephalic fetuses with suspected
24. Bueno MG, Martinez N, Abdalla L, et al: Animals in the Zika congenital Zika virus syndrome, Ultrasound Obstet Gynecol
virus life cycle: what to expect from megadiverse Latin American 50(6):717–722, 2017.
countries, PLoS Negl Trop Dis 10:2016. Available at: http:// 43. Schuler-Faccini L, Ribeiro EM, Feitosa IML, et al: Possible asso-
www.ncbi.nlm.nih.gov/pmc/articles/PMC5179043/. (Accessed ciation between Zika virus infection and microcephaly - Brazil,
29 January 2017). 2015, MMWR Morb Mortal Wkly Rep 65:59–62, 2016.
25. Dudley DM, Aliota MT, Mohr EL, et al: A rhesus macaque 44. Carvalho CAM, Casseb SMM, Gonçalves RB, et al: Bovine lac-
model of Asian-lineage Zika virus infection, Nat Commun toferrin activity against Chikungunya and Zika viruses, bioRxiv
7:12204, 2016. 071571, 2016.
42
An Overview of Middle East
Respiratory Syndrome in the
Middle East
ARSHAD HAROON TOOSY AND SEAN O’SULLIVAN
Introduction
is a positive-sense enveloped single-stranded RNA virus and
Middle East respiratory syndrome (MERS) is an emerging is the first lineage of 2c Betacoronavirus known to infect
infectious zoonotic disease caused by a novel coronavirus humans.2,8 It is more closely related to bat CoVs HKU4
(CoV). MERS was first reported in 2012 in Jeddah, and HKU5 (lineage 2c) than to the severe acute respiratory
Kingdom of Saudi Arabia (KSA), and in Jordan, respec- syndrome CoV (SARS-CoV, (lineage 2b).2,3 Recent genome
tively.1 The disease was considered a potential pandemic sequencing analysis reported the genomic evolution rate
threat to public health in the Persian Gulf region.2 (See (1.12 × 103 substitutions per site), suggesting that MERS-
also Chapter 19.) CoV diverged from its viral ancestor in March 2012.12
Most known CoVs infect and circulate in animals, mainly Analysis of human MERS-CoV sequences has identi-
bats, but a number of CoVs are known to cause human fied several circulating genotypes. These distinct genotypes
disease (see also Chapter 40).3–5 The rapid emergence of are phylogenetically classified into clades A, B, and, most
MERS-CoV coupled with its limited geographic distribu- recently, C, which correlate with outbreaks of MERS
tion has led to the suspicion that this is a zoonotic disease among humans.4,5,8,12 The emergence of divergent MERS-
with an animal reservoir,4 and the evidence supports the CoV clades in humans since 2012 is consistent with several
hypothesis that dromedary camels (DCs) are the reservoir independent sporadic introductions into the human popu-
host. In DCs MERS-CoV causes a mild, transient upper lation from an animal reservoir, of which the camel was
respiratory tract (URT) infection.4–6 unquestionably the source.6,8,12,13
A mild or asymptomatic disease has also been reported in
humans, but this is not always the case. MERS-CoV infec- Pathogenesis
tion in humans often results in a severe, life-threatening
disease of the lower respiratory tract (LRT), with high mor- Host cell entry of MERS-CoV is mediated by the binding
tality.1,2,5 Immunocompromised, elderly people and those of MERS spike (S) proteins to a specific cellular recep-
with comorbidities, usually with a history of close contact tor known as dipeptidyl peptidase 4 (DPP4).11 DPP4 is
with infected DCs, are particularly susceptible.7,8 In such expressed on the epithelial and endothelial cells of most
cases the disease progresses rapidly, resulting in multiorgan human organ tissues in ex vivo studies using human tissue
failure and acute respiratory distress syndrome. Between culture lines; this may account for the multisystem clinical
2012 and April 7, 2017, a total of 1936 confirmed human spectrum of the MERS-CoV infection.2,14
cases were reported to the World Health Organization A strain cultured from a fatal human case was experi-
(WHO), resulting in 690 fatalities (crude case fatality rate mentally inoculated into three DCs using intratracheal,
of 36%; see Fig. 42.1).1,9 Potential clinical cases of MERS intranasal, and conjunctival routes.15 A mild transient
in DCs must be reported to the World Organisation for disease resulted in submucosal inflammation and necrosis
Animal Health (OIE) as an emerging infectious disease.10 in the URT and LRT, but the alveoli remained unaffected.15
Experimental inoculation of rhesus macaques (Macaca
Virology mulatta) and common marmosets (Callithrix jacchus)
resulted in mild to severe LRT disease causing multifocal
MERS-CoV is a member of the subfamily Coronavirinae, interstitial pneumonia in the macaques and extensive fatal
genus Betacoronavirus, subgroup lineage 2c.11 MERS-CoV pneumonia in the marmosets.2,14,16
287
288 SE C T I O N 8 Emerging and Changing Infectious Diseases
• Figure 42.1 Epicurve: global confirmed cases of Middle East respiratory syndrome–coronavirus
(MERS-CoV), reported to the World Health Organization as of April 7, 2017 (n = 1936). (Reprinted from
WHO Emergencies: MERS-CoV. <http://www.who.int/emergencies/mers-cov/en/>.)
strategic serosurveys of humans using samples collected Genetic deep sequencing methods (i.e., high-throughput
after 2012 have been infrequent.4,5,10 There is a paucity of sequencing) have been readily available to researchers since
baseline data to describe the proportion of the potentially the disease was first reported.12 Sequenced data have been
infected human population for much of the Arabian Penin- used in these cases to successfully construct the phylogenetic
sula and all of East Africa, including the Horn of Africa.10 tree between related viruses and hosts.2,6,7,12
Direct MERS-CoV antigen detection is possible but has
Transmission been rarely performed.10 Immunochromatography assays
and monoclonal antibody-based capture ELISAs targeting
The exact mechanism of transmission from camels to the MERS-CoV nucleocapsid protein have been described.20
human remains uncertain.10 Sustained close contact is most Since the virus was first reported in 2012, a range of
probably necessary for transmission by aerosolized droplets, comprehensive laboratory tests has been developed.10,30
as MERS-CoV viral RNA has been detected in air samples To better understand the disease, it has been important
from a barn housing infected DCs in Qatar, and the virus to collate sampling methodology data, laboratory results,
may remain viable in aerosol for up to 45 minutes.10,27–29 The and analyses in combination with clinical and epidemio-
potential public health risk resulting from aerosol-generating logic data.10 Until laboratory assays are fully validated, a
activities ranges from contamination of a room occupied by combination of molecular and serologic laboratory tests
a symptomatic patient to slaughter practices.8,24,30 is required to improve confidence in laboratory diagnosis
Aerosolized transmission of MERS-CoV has been attrib- during outbreaks.30 In cases of mild or asymptomatic
uted to hospital outbreaks in KSA and South Korea.26,27,29 infection, full validation of serologic assays is required to
MERS-CoV spreads inefficiently from human to human, rule out false-negative results.31 Validation is also required
but transmission is effective in a hospital environment, to successfully apply newly developed diagnostic serology
where susceptible individuals are concentrated and the algorithms to inform public health decisions.10,30,31
risks are amplified by poor infection prevention and control
(IPC) protocols.8 Treatment
In some reported cases of MERS, direct contact with
camels was not apparent.27,29 Camel-to-human transmission Therapeutic options for MERS-CoV are limited. Sup-
through other routes is, however, possible owing to the portive treatment is indicated for hospitalized patients, but
consumption of unpasteurized camel milk or raw camel vigilance for complications is essential.8 Empirical use of
meat and in traditional medicine, when camel urine is antimicrobial agents or steroids has not succeeded in revers-
consumed as a natural remedy for a variety of ailments.3,10 ing the progression of severe disease.8,27,29 No specific drug
A recent survey has found that infected camels may shed or vaccine is currently available to treat MERS. Indeed,
MERS-CoV virus in milk and urine, and the virus has been it has been stated that the complexity and time required
shown to remain infectious for 3 days in milk stored at for the development and registration process of drugs for
4°C.3,10 The transmission risks associated with the handling human use impedes the ability to counter the rapid threat
of camel products, raw milk, urine, and meat during animal against an emerging infectious agent.8 For example, there
slaughter are yet to be fully elucidated. Further studies are is no vaccine available against SARS-CoV because of the
needed to demonstrate the potential of camel-to-human brevity of the threat to the public health.2,8 It is likely that
transmission.8 a MERS-CoV vaccine for human use may not be developed
due to a lack of commercial interest, or if the threat posed
Diagnosis by MERS-CoV declines in the meantime.8
Nevertheless, given the prevalence of MERS-CoV infec-
Serologic methods with high sensitivity and specificity to tion in the Middle East’s DC population and due to the
detect MERS-CoV antibodies have been developed for potential for spillover to the human population in direct
use in seroepidemiologic studies. Methods include indirect contact with DCs, the development of a vaccine for use in
immunofluorescence assays, enzyme-linked immunosor- DCs may be feasible.4,5,32 A recent successful trial of a MERS
bent assays (ELISAs), protein microarray technology, and orthopoxvirus vaccine has conferred mucosal immunity in
microneutralization (MN) assays.13,20–22 Pseudoparticle the URTs of DCs.32 Eradication of MERS-CoV from herds
virus neutralization tests (ppNTs) and conventional MN may be possible, if vaccines are administered to young,
assays have also been used to detect neutralizing antibodies immunologically naive camels prior to exposure.4,5,32
to MERS-CoV.18
Validated molecular assays have been developed.10,13,19 Control and Prevention
RT-rtPCR is the preferred diagnostic method for the detec-
tion of MERS-CoV and has been endorsed by the WHO.1 Identification of the zoonotic source of MERS guides control
Confirmation of MERS in suspected cases requires the strategies at the human-animal interface.3,30 By preventing
screening of samples targeting a number of genes specific spillover of MERS-CoV from animals to humans, the risk
to MERS-CoV, namely Up E, ORF 1a, ORF 1b, and N of nosocomial and familial outbreaks in the Middle East
genes.1,10,13,19 could be eliminated.3
290 SE C T I O N 8 Emerging and Changing Infectious Diseases
At present the implementation of intensive IPC mea- its infancy. Aside from bats, the role that other wildlife may
sures in human health care is vital, including improving play in the ecology of MERS-CoV in East Africa and Arabia
education and awareness among healthcare workers.1,8 Most is yet to be elucidated.
human cases have been linked to lapses in IPC, as one-fifth
of viral infections have been reported among healthcare Acknowledgments
workers.1,8 Stringent precautions while handling suspected
MERS-CoV patients include the use of personal protective The authors wish to thank the following: H. E. Ghanim
equipment (PPE) (i.e., disposable gloves, gowns, respiratory Mubarak Al Hajeri and the senior management of Al Ain
protection, and eye protection).2,8,33 Immunocompromised Zoo for their encouragement and support, Dr. Ahsan Ul
individuals and those with preexisting medical conditions Haq of Dubai Camel Hospital for his technical insight,
should avoid close contact with DCs.2,8 and Dr. Andrew Higgins, Fellow of the Zoological Society
Public health authorities should adopt a standardized of London and Honorary editor in chief of The Veterinary
public health response protocol to include standardized Journal, who reviewed the manuscript.
case reporting methodology as defined by the WHO.1,30
Standardization of case definitions aids accurate calculation References
of a case fatality ratio by including mild or asymptomatic
cases.30 The Health Authority of Abu Dhabi in the United 1. WHO MERS-CoV Global Summary and risk assessment, 5
December 2016 WHO/MERS/RA/16.1. http://www.who.int/
Arab Emirates recently implemented a standardized report-
emergencies/mers-cov/mers-summary-2016.pdf?ua=1. (Accessed
ing option for MERS, successfully incorporating it into 1 March 2017).
existing epidemiologic surveillance systems with the aim of 2. Banik GR, Khandaker G, Rashid H: Middle East respiratory
enhancing surveillance, educating healthcare workers, and syndrome coronavirus “MERS-CoV”: current knowledge gaps,
ensuring laboratory capacity.25 Paediatr Respir Rev 16(3):197–202, 2015.
In countries where MERS-CoV is enzootic in DCs, 3. Han H-J, Yu H, Yu X-J: Evidence for zoonotic origins of
MERS control at the animal-human interface is unlikely Middle East respiratory syndrome coronavirus, J Gen Virol
to succeed unless appropriate preventive strategies are 97(2):274–280, 2016.
implemented.5 These should include the following: 4. Mohd H, Al-Tawfiq J, Memish Z: Middle East respiratory
• Strict regulation of camel movement with imposition syndrome coronavirus (MERS-CoV) origin and animal reservoir,
of a requirement for MERS clearance prior to the Virol J 13(1):87, 2016.
5. Omrani A, Al-Tawfiq J, Memish Z: Middle East respiratory
importation and transport of camels, including animals
syndrome coronavirus (MERS-CoV): animal to human interac-
presented for slaughter. tion, Pathog Glob Health 109(8):354–362, 2015.
• Camels with detectable MERS-CoV RNA should be 6. Haagmans B, Al Dhahiry S, Reusken C, et al: Middle East respi-
quarantined and tested at regular intervals. ratory syndrome coronavirus in dromedary camels: an outbreak
• Use of appropriate PPEs while handling DCs. investigation, Lancet Infect Dis 14(2):140–145, 2014.
• Increased awareness among camel owners and the general 7. Azhar E, El-Kafrawy S, Farraj S, et al: Evidence for camel-to-human
public of the risks of consuming unpasteurized camel transmission of MERS coronavirus, NEJM 370(26):2499–2505,
milk and urine. This may prove challenging given the 2014.
depth of customs and beliefs in some areas. 8. Mackay IM, Arden KE: Middle East respiratory syndrome: an
• Accelerated development of safe and effective MERS emerging coronavirus infection tracked by the crowd, Virus Res
vaccines for animal and human use.5,33 202:60–88, 2015.
9. WHO Emergencies: MERS-CoV. Available at: http://
www.who.int/emergencies/mers-cov/en/. (Accessed 8 April
Conclusions 2017).
10. Mackay IM, Arden KE: MERS coronavirus: diagnostics, epide-
MERS-CoV has been observed for only 4 years, and vigilance miology and transmission, Virol J 12(1):222, 2015.
is vital for the containment of the disease due to the high 11. Raj VS, Mou H, Smits SL, et al: Dipeptidyl peptidase 4 is a
case fatality rate in humans and possible genetic instability functional receptor for the emerging human coronavirus-EMC,
of the virus.8 Continued laboratory testing, genetic sequenc- Nature 495:251–254, 2013.
ing, analysis, timely data sharing, and clear communication 12. Cotten M, Lam TT, Watson SJ, et al: Full-genome deep sequenc-
are essential if such vigilance is to be effective.8 Nonetheless, ing and phylogenetic analysis of novel human betacoronavirus,
despite the potential for a pandemic outbreak at multiple Emerg Infect Dis 19:736–42B, 2013.
mass gatherings during the Islamic calendar (Hajj, Eid, and 13. Alagaili A, Briese T, Mishra N, et al: Middle East respiratory
syndrome coronavirus infection in dromedary camels in Saudi
Umrah) there were no reported outbreaks of MERS during
Arabia, MBio 5(2):e00884–14, 2014.
or immediately after these events.10 As such MERS-CoV is 14. van den Brand J, Smits S, Haagmans B: Pathogenesis of Middle
not a virus of pandemic concern.10 East respiratory syndrome coronavirus, J Pathol 235:175–184,
Since 2012 our understanding of MERS has increased 2015, doi:10.1002/path.4458.
greatly although gaps in knowledge still exist. The under- 15. Adney DR, van Doremalen N, Brown VR, et al: Replication and
standing of the disease’s ecology—especially the interplay shedding of MERS-CoV in upper respiratory tract of inoculated
between camels, humans, and the environment—is still in dromedary camels, Emerg Infect Dis 20:1999–2005, 2014.
CHAPTER 42 An Overview of Middle East Respiratory Syndrome in the Middle East 291
16. Yao Y, Bao L, Deng W, et al: An animal model of MERS pro- 25. Al Hosani FI, Pringle K, Al Mulla M, et al: Response to emer-
duced by infection of rhesus macaques with MERS coronavirus, gence of Middle East respiratory syndrome coronavirus, Abu
J Infect Dis 209(2):236–242, 2014. Dhabi, United Arab Emirates, 2013–2014, Emerg Infect Dis
17. Anthony SJ, Gilardi K, Menachery VD, et al: Further evidence 22(7):1162, 2016.
for bats as the evolutionary source of Middle East respiratory 26. Hunter J, Nguyen B, Aden D, et al: Transmission of Middle
syndrome coronavirus, MBio 8:e00373–17, 2017. https:// East respiratory syndrome coronavirus infections in healthcare
doi.org/10.1128/mBio.00373-17. (Accessed 18 April 2017). settings, Abu Dhabi, Emerg Infect Dis 22(4):647, 2016.
18. Memish Z, Mishra N, Olival K, et al: Middle East respiratory 27. Assiri A, McGeer A, Perl TM, et al: Hospital outbreak of Middle
syndrome coronavirus in bats, Saudi Arabia, Emerg Infect Dis East respiratory syndrome coronavirus, NEJM 369:407–416,
19(11):2013. 2013.
19. Chu D, Poon L, Gomaa N, et al: MERS coronaviruses in 28. Azhar EI, Hashem AM, El-Kafrawy SA, et al: Detection of the
dromedary camels, Emerg Infect Dis 20(6):1049–1053, 2014. Middle East respiratory syndrome coronavirus genome in an
20. Hemida M, Perera R, Wang P, et al: Middle East respiratory air sample originating from a camel barn owned by an infected
syndrome (MERS) coronavirus seroprevalence in domestic live- patient, MBio 5:e1450–e1514, 2014.
stock in Saudi Arabia, 2010 to 2013, Euro Surveill 18(50), 2013. 29. Oboho I, Tomczyk S, Al-Asmari A, et al: 2014 MERS-CoV
pii=20659. Retrieved from http://dx.doi.org/10.2807/1560 outbreak in Jeddah—a link to health care facilities, NEJM
-7917.ES2013.18.50.20659. (Accessed 1 March 2017). 372(9):846–854, 2015.
21. Meyer B, Müller M, Corman V, et al: Antibodies against MERS 30. de Sousa R, Reusken C, Koopmans M: MERS coronavirus:
coronavirus in dromedaries, United Arab Emirates, 2003 and data gaps for laboratory preparedness, J Clin Virol 59(1):4–11,
2013, Emerg Infect Dis 20(4):552–559, 2014. 2014.
22. Reusken C, Ababneh M, Raj V, et al: Middle East respiratory 31. Al Hammadi ZM, Chu DK, Eltahir YM, et al: Asymptomatic
syndrome coronavirus (MERS-CoV) serology in major livestock MERS-CoV infection in humans possibly linked to infected
species in an affected region in Jordan, June to September 2013, camels imported from Oman to United Arab Emirates, Emerg
Euro Surveill 18(50), 2013. pii=20662. Retrieved from http:// Infect Dis 21:12, 2015.
www.eurosurveillance.org/ViewArticle.aspx?ArticleId=20662. 32. Haagmans B, van den Brand J, Raj V, et al: An orthopoxvirus-
(Accessed 1 March 2017). based vaccine reduces virus excretion after MERS-CoV infection
23. Raj V, Farag E, Reusken C, et al: Isolation of MERS Corona- in dromedary camels, Science 351(6888):77–81, 2016.
virus from a Dromedary Camel, Qatar, Emerg Infect Dis 20(8), 33. Centers for Disease Control and Prevention: Interim prevention
2014. and control recommendations for hospitalized patients with
24. Khalafalla A, Lu X, Al- Mubarak A, et al: MERS-CoV in upper Middle East respiratory syndrome coronavirus (MERS-CoV).
respiratory tract and lungs of dromedary camels, Saudi Arabia, Available at: https://www.cdc.gov/coronavirus/mers/infection-
2013–2014, Emerg Infect Dis 21(7):1153, 2015. prevention-control.html. (Accessed 18 April 2017).
43
Disease Risk to Endemic Animals From
Introduced Species on Madagascar
FIDISOA RASAMBAINARIVO AND STEVEN M. GOODMAN
292
CHAPTER 43 Disease Risk to Endemic Animals From Introduced Species on Madagascar 293
elsewhere in the world, the reason for the observed declines sparrow [Passer domesticus] and house crow [Corvus splen-
is likely a combination of factors, including resource com- dens]). These nonnative birds may facilitate the introduc-
petition, predation, and disease.20,21 This chapter attempts tion of pathogens and pose different threats to the native
to review the literature on pathogen introduction from Malagasy avifauna, as has been the case for bird populations
exotic hosts and highlights the risks of disease introduction elsewhere.10,11 For example, in the Galápagos, it is thought
on the rich and unique fauna of Madagascar. that the introduction of the avipoxvirus and Haemoproteus
blood pathogens in the 19th century was associated with
either pet caged birds or natural songbird migration.32
Disease Risks From Some species of Haemoproteus are known to reduce the
Introduced Amphibians physical condition and reproductive success of captive and
wild birds; on Madagascar, this genus and Plasmodium
The Asian common toad (Duttarphrynus melanosticus) are transmitted by numerous species of mosquitoes living
was first reported in Madagascar in 2014.22 This invasive sympatrically with native birds.33,34 Most studies of bird
alien amphibian species is widely distributed across many pathogens on Madagascar focus on domestic fowl, and only
environmental types in Asia and may competitively exclude a few investigations have documented pathogens infecting
Malagasy amphibians. It has also been hypothesized that the island’s native birds.33,35
these toads might affect Madagascar’s native carnivorans West Nile virus (WNV), a member of the family Fla-
through their cardiotoxic toxins.23 viviridae, is widely distributed in parts of the Old World.
An additional concern regarding this invasive toad During the summer of 1999, this virus was introduced in
species is the cointroduction of pathogens, particularly the Western Hemisphere presumably by the transport of
Ranavirus and the amphibian chytrid fungus Batracho- infected humans, birds, or mosquitoes and spread rapidly,
chytridium dendrobatidis (Bd). Both have been associated causing morbidity and mortality of birds and mammals in
with major declines in amphibian populations elsewhere in the United States and Canada.36,37 On Madagascar, evidence
the world.24,25 Madagascar was previously considered to be of WNV exposure was found in both introduced and native
one of the last major land masses without this amphibian animal species including birds, peridomestic rodents, fruit
fungal pathogen.26 Despite several surveys and long-term bats (Pteropus rufus), and several lemur species.31,38–40
research, the presence of the pathogen was not confirmed However, the phylogenetics of the WNV strains isolated
on Madagascar before 2010.26,27 More recently, the fungus on Madagascar suggest a local WNV transmission cycle
was detected on several amphibians shipped from Madagas- with no new, recent introductions of WNV despite the
car to the United States.28 Subsequently, and as a result of migration and introduction of several bird and arthropod
coordinated efforts, the pathogen was found in wild frogs species on the island.31,39 The introduction to Madagascar
in multiple areas across the island, where representatives of of a foreign strain of WNV might have a dramatic effect
all anuran families have tested Bd-positive.26,28 However, on the local fauna.
results from field surveys from the same sites are conflicting, Similarly, Newcastle disease virus (NDV), a member of
and there has been no evidence of mass amphibian mortali- the Paramyxoviridae, is distributed throughout the world
ties, raising the question of whether the pathogen is firmly and infects a wide range of birds. Virulent forms of NDV
established or is recurrently introduced.29 cause widespread and highly contagious disease in both
In its native range, the Asian common toad is also known domestic and wild birds.41 It is responsible for the high mor-
to harbor potentially zoonotic pathogens such as Salmo- tality of chickens in rural Madagascar, which in turn causes
nella30; by extrapolation, its introduction to Madagascar important economic burdens.42 In most cases, domestic
may also become a concern for public health or the native bird infections on the island implicate genotype XI of
Malagasy fauna. In any case, further research is needed NDV, a form related to the variant responsible for the first
regarding the pathogens that this toad may carry. pandemic of Newcastle disease (genotype IV), whereas wild
birds are primarily infected by genotype Ib, although some
Disease Risks From Introduced Birds are also infected by genotype XI.43,44 These results suggest
that genotype XI circulates across Madagascar between wild
Approximately 209 of the 283 bird species found on Mada- and domestic birds and may potentially affect either the
gascar are breeding residents and a few have regular seasonal native or exotic bird populations via spillover and spillback
migration between sub-Saharan Africa and islands in the mechanisms.45
western Indian Ocean. The number of Palearctic migrants, A recent study has disclosed that certain coronaviruses,
particularly Passeriformes, passing through Madagascar is specifically of the genus Gammacoronavirus, are present in
very limited compared with continental Africa; this has wild Malagasy birds, particularly species living in aquatic
important implications for the potential transmission of habitats.46 Coronaviruses are found in a variety of animals,
pathogens between birds.31 In addition, several species have in which they can cause respiratory, enteric, and neurologic
been introduced either intentionally for food production diseases. This particular genus is associated with infectious
(e.g., poultry and other domestic fowl) or pest control bronchitis virus, but birds that tested positive did not appear
(common myna [Acridotheres tristis]) or accidentally (house to be affected.46 The strains of avian coronaviruses detected
294 SE C T I O N 8 Emerging and Changing Infectious Diseases
The spirurid parasite Spirocerca lupi is another pathogen rattus.69 Members of the Paramyxoviridae viral family have
transmitted by introduced dogs that may negatively affect been associated with a number of emerging diseases that
the native fauna of Madagascar.58 This nematode parasite affect humans and natural animal populations. Evidence
is prevalent in free-roaming rural dogs on the island and indicates that R. rattus is an important spreader of UMRVs
is presumably responsible for the death of several captive and that there is considerable lateral exchange of UMRVs
lemurs through the formation of aortic aneurysms and their between sympatrically occurring mammals.69 To our knowl-
subsequent rupture.59 Whether these parasites negatively edge, no data are available on the isolation of Morbillivirus
affect endemic population of lemurs and carnivorans is among lemurs and carnivorans on Madagascar.
unknown and warrants further research.
Additionally, it has recently been found that some brown Conclusions
lemurs (Eulemur albifrons) had antibodies to Toxoplasma
gondii, a protozoan whose only known definitive hosts are Introduced species are an increasingly dominant part of
members of the family Felidae, represented on Madagascar many natural and human-modified landscapes. It is esti-
by introduced cats.60 This indicates a disease spillover from mated that invasive species, particularly invasive mammal
exotic felids to the endemic mammalian fauna of the island. predators such as dogs and cats, have caused the extinction
Similarly, 95% (n = 44) of the Cryptoprocta evaluated in two of at least 87 species of birds, 45 species of mammals, and
western protected areas had antibodies against T. gondii.57 10 species of reptiles and that they are threatening many
This parasite was associated with neurologic disease and more.70 Some of these extinctions may be mediated by the
death in several captive Malagasy species and may potentially introduction of pathogens.71 On Madagascar, several of the
affect wild endemic euplerid and lemur populations.61,62 “worst invasive species” were introduced following human
colonization.72 Their impacts, including those of associated
Disease Risks From Introduced Rodents pathogens, on the fauna of Madagascar warrant further
research and monitoring. It is hoped that the data generated
Pathogenic genospecies of Leptospira bacteria were also from such research will ultimately pave the way to successful
detected in a large proportion of introduced rodents in strategies for managing the risks of “pathogen pollution”
various parts of Madagascar, notably Leptospira interrogans in and disease spillover to the native fauna of Madagascar.
Rattus rattus.63 Leptospira interrogans is of particular concern
because it is known to be pathogenic in more than 150 Acknowledgments
animal species.64 Infected Rattus, then, constitute a threat to
both public health and animal conservation.64,65 One of the The authors would like to extend thanks to Ingrid Porton,
interesting results of these studies is the genetic uniqueness Franco Andreone, and Goncalo Rosa for their contributions
of several Leptospira forms among native Malagasy terres- to this chapter.
trial and volant mammals, indicating probable isolation
and evolution in deep time.66 This suggests little exchange
of Leptospira between Rattus and native small mammals.66 References
Exposure of other native species including lemurs and
carnivorans to Leptospira interrogans from invasive rodents 1. Lowe S, Browne M, Boudjelas S, et al: 100 of the world’s worst
is unknown. invasive alien species: A selection from the global invasive species
In a similar pattern, native rodents seem to harbor differ- database. The Invasive Species Specialist Group (ISSG) of the
Species Survival Commission (SSC) of the World Conservation
ent species of trypanostomid parasites than their introduced
Union (IUCN), 2000.
counterparts. In the Ranomafana National Park, 30% 2. Hughes J, Macdonald DW: A review of the interactions
of R. rattus examined during a survey were infected by between free-roaming domestic dogs and wildlife, Biol Conserv
Trypanosoma lewisii, whereas the native and sympatric neso- 157:341–351, 2013.
myines belonging to the genus Eliurus were infected with 3. Cunningham A, Daszak P, Rodriguez J: Pathogen pollution:
a morphologically different trypanostomid.67 The authors Defining a parasitological threat to biodiversity conservation, J
suggest that native rodents may not be infected by the Parasitol 89:S78–S83, 2003.
parasite from introduced species or that the relatively recent 4. Kelly DW, Paterson RA, Townsend CR, et al: Parasite spillback:
invasion of Rattus in the interior of the park has not yet A neglected concept in invasion ecology? Ecology 90:2047–2056,
resulted in the transmission of these parasites. Elsewhere, on 2009.
Christmas Island, Trypanosoma lewisii carried by R. rattus 5. Prenter J, MacNeil C, Dick J, et al: Roles of parasites in animal
invasions, Trends Ecol Evol (Amst) 19:385–390, 2004.
was associated with the infection and subsequent decline of
6. Holt RD, Grover J, Tilman D: Simple rules for interspecific
R. macleari, a native rodent.68 dominance in systems with exploitative and apparent competi-
Recent research on Madagascar has found a virus of the tion, Am Nat 741–771, 1994.
genus Morbillivirus that is currently considered as unclas- 7. Rushton S, Lurz P, Gurnell J, et al: Modelling the spatial dynam-
sified Morbilli-related paramyoxoviruses (UMRVs).69 This ics of parapoxvirus disease in red and grey squirrels: A possible
virus is thought to be widespread among endemic rodents, cause of the decline in the red squirrel in the UK? J Appl Ecol
tenrecs, and bats as well as introduced rodents, specifically R. 37:997–1012, 2000.
296 SE C T I O N 8 Emerging and Changing Infectious Diseases
8. Medina FM, Bonnaud E, Vidal E, et al: A global review of the 29. Kolby JE, Skerratt LF: Amphibian chytrid fungus in Madagascar
impacts of invasive cats on island endangered vertebrates, Glob neither shows widespread presence nor signs of certain establish-
Chang Biol 17:3503–3510, 2011. ment, PLoS ONE 10:2015.
9. Mooney HA, Cleland EE: The evolutionary impact of invasive 30. Ribas A, Poonlaphdecha S: Wild-caught and farm-reared
species, Proc Natl Acad Sci USA 98:5446–5451, 2001. amphibians are important reservoirs of salmonella, a study in
10. van Riper C, van Riper SG, Goff ML, et al: The epizootiology north-east Thailand, Zoonoses Public Health 2016.
and ecological significance of malaria in Hawaiian land birds, 31. Tantely ML, Goodman SM, Rakotondranaivo T, et al: Review
Ecol Monogr 56:327–344, 1986. of West Nile virus circulation and outbreak risk in Madagascar:
11. van Riper C, van Riper SG, Hansen WR, et al: Epizootiology and Entomological and ornithological perspectives, Parasite 23:2016.
effect of avian pox on Hawaiian forest birds, Auk 119:929–942, 32. Parker PG, Buckles EL, Farrington H, et al: 110 years of avipox-
2002. virus in the Galapagos Islands, PLoS ONE 6:2011.
12. Myers N, Mittermeier RA, Mittermeier CG, et al: Biodiversity 33. Schmid S, Dinkel A, Mackenstedt U, et al: Avian malaria on
hotspots for conservation priorities, Nature 403:853–858, Madagascar: Bird hosts and putative vector mosquitoes of dif-
2000. ferent Plasmodium lineages, Parasit Vectors 10:6, 2017.
13. Poux C, Madsen O, Marquard E, et al: Asynchronous coloniza- 34. Merino S, Moreno J, Sanz JJ, et al: Are avian blood parasites
tion of Madagascar by the four endemic clades of primates, pathogenic in the wild? A medication experiment in blue tits
tenrecs, carnivores, and rodents as inferred from nuclear genes, (Parus caeruleus), Proc R Soc Lond B Biol Sci 267:2507–2510,
Syst Biol 54:719–730, 2005. 2000.
14. Jenkins RK, Tognelli MF, Bowles P, et al: Extinction risks and 35. Savage AF, Robert V, Goodman SM, et al: Blood parasites in
the conservation of Madagascar’s reptiles, PLoS ONE 9:e100173, birds from Madagascar, J Wildl Dis 45:907–920, 2009.
2014. 36. McLean R, Ubico S, Bourne D, et al: West Nile virus in livestock
15. Vieites DR, Wollenberg KC, Andreone F, et al: Vast underesti- and wildlife. In Mackenzie J, Barrett ADT, Deubel V, editors:
mation of Madagascar’s biodiversity evidenced by an integrative Japanese encephalitis and West Nile viruses, Berlin, 2002, Springer,
amphibian inventory, Proc Natl Acad Sci USA 106:8267–8272, pp 271–308.
2009. 37. Lanciotti R, Roehrig J, Deubel V, et al: Origin of the West
16. Glaw F, Vences M: A field guide to the amphibians and reptiles of Nile virus responsible for an outbreak of encephalitis in the
Madagascar, ed 3, Cologne, 2007, Vences & Glaw. northeastern United States, Science 286:2333–2337, 1999.
17. Dewar RE, Richard AF: Madagascar: A history of arrivals, 38. Sondgeroth K, Blitvich B, Blair C, et al: Assessing flavivirus,
what happened, and will happen next, Annu Rev Anthropol lentivirus, and herpesvirus exposure in free-ranging ring-tailed
41:495–517, 2012. lemurs in southwestern Madagascar, J Wildl Dis 43:40–47, 2007.
18. Goodman SM: Rattus on Madagascar and the dilemma of 39. Maquart M, Boyer S, Rakotoharinome VM, et al: High preva-
protecting the endemic rodent fauna, Conserv Biol 9:450–453, lence of West Nile virus in domestic birds and detection in 2 new
1995. mosquito species in Madagascar, PLoS ONE 11:e0147589, 2016.
19. Farris ZJ, Golden CD, Karpanty S, et al: Hunting, exotic 40. Root JJ: West Nile virus associations in wild mammals: A syn-
carnivores, and habitat loss: Anthropogenic effects on a native thesis, Arch Virol 2012.
carnivore community, Madagascar, PLoS ONE e0136456, 41. Alexander D, Senne D: Newcastle disease, other avian paramyxo-
2015. viruses, and pneumovirus infections. In Diseases of poultry, ed 12,
20. Vanak AT, Gompper ME: Interference competition at the Ames, IA, 2008, Blackwell Publishing, pp 75–98.
landscape level: The effect of free-ranging dogs on a native 42. Maminiaina OF, Koko M, Ravaomanana J, et al: Epidemiology
mesocarnivore, J Appl Ecol 47:1225–1232, 2010. of Newcastle disease in village poultry farming in Madagascar,
21. Knobel DL, Butler JR, Lembo T, et al: Dogs, disease, and wildlife. Rev - Off Int Epizoot 26:691–700, 2007.
In Gompper ME, editor: Free-ranging dogs and wildlife conserva- 43. de Almeida RS, Hammoumi S, Gil P, et al: New avian paramyxo-
tion, Oxford, 2014, Oxford University Press, pp 144–169. viruses type I strains identified in Africa provide new outcomes
22. Kolby JE, Kraus F, Rabemananjara F, et al: Stop Madagascar’s for phylogeny reconstruction and genotype classification, PLoS
toad invasion now, Nature 509:563–564, 2014. ONE 8:2013.
23. Brown KA, Farris ZJ, Yesuf G, et al: Modeling co-occurrence 44. Maminiaina OF, Gil P, Briand F-X, et al: Newcastle disease virus
between toxic prey and naïve predators in an incipient invasion, in Madagascar: Identification of an original genotype possibly
Biodivers Conserv 25:2723–2741, 2016. deriving from a died out ancestor of genotype iv, PLoS ONE
24. Andreone F: Madagascar: Risk review is under way for invasive 5:2010.
toad, Nature 512:253, 2014. 45. Cappelle J, Caron A, De Almeida RS, et al: Empirical analysis
25. Daszak P, Berger L, Cunningham AA, et al: Emerging infectious suggests continuous and homogeneous circulation of Newcastle
diseases and amphibian population declines, Emerg Infect Dis disease virus in a wide range of wild bird species in Africa,
5:735, 1999. Epidemiol Infect 143:1292, 2015.
26. Kolby JE: Presence of the amphibian chytrid fungus batracho- 46. de Sales Lima FE, Gil P, Pedrono M, et al: Diverse gammacoro-
chytrium dendrobatidis in native amphibians exported from naviruses detected in wild birds from Madagascar, Eur J Wildl
Madagascar, PLoS ONE 9:2014. Res 61:635–639, 2015.
27. Andreone F, Carpenter AI, Cox N, et al: The challenge of 47. Köndgen S, Kühl H, N’Goran PK, et al: Pandemic human viruses
conserving amphibian megadiversity in Madagascar, PLoS Biol cause decline of endangered great apes, Curr Biol 18:260–264,
6:943–946, 2008. 2008.
28. Bletz MC, Rosa GM, Andreone F, et al: Widespread presence 48. Junge RE, Barrett MA, Yoder AD: Effects of anthropogenic
of the pathogenic fungus Batrachochytrium dendrobatidis in wild disturbance on indri (Indri indri) health in Madagascar, Am J
amphibian communities in Madagascar, Sci Rep 5:8633, 2015. Primatol 73:632–642, 2011.
CHAPTER 43 Disease Risk to Endemic Animals From Introduced Species on Madagascar 297
49. Irwin MT, Junge RE, Raharison J-LL, et al: Variation in physi- 61. Spencer JA, Joiner KS, Hilton CD, et al: Disseminated toxo-
ological health of diademed sifakas across intact and fragmented plasmosis in a captive ring-tailed lemur (Lemur catta), J Parasitol
forest at Tsinjoarivo, eastern Madagascar, Am J Primatol 90:904–906, 2004.
72:1013–1025, 2010. 62. Corpa J, García-Quirós A, Casares M, et al: Encephalomyelitis
50. Rasambainarivo F, Gillespie T, Wright P, et al: Survey of giardia by Toxoplasma gondii in a captive fossa (Cryptoprocta ferox), Vet
and cryptosporidium in lemurs from the Ranomafana National Parasitol 193:281–283, 2013.
Park, Madagascar, J Wildl Dis 49:741–743, 2013. 63. Rahelinirina S, Léon A, Harstskeerl RA, et al: First isolation
51. Villers LM, Jang SS, Lent CL, et al: Survey and comparison and direct evidence for the existence of large small-mammal
of major intestinal flora in captive and wild ring-tailed lemur reservoirs of Leptospira sp. In Madagascar, PLoS ONE 5:e14111,
(Lemur catta) populations, Am J Primatol 70:175–184, 2008. 2010.
52. Bodager JR, Parsons MB, Wright PC, et al: Complex epidemi- 64. Bharti AR, Nally JE, Ricaldi JN, et al: Leptospirosis: A zoonotic
ology and zoonotic potential for Cryptosporidium suis in rural disease of global importance, Lancet Infect Dis 3:757–771,
Madagascar, Vet Parasitol 207:140–143, 2015. 2003.
53. Charles-Smith LE, Cowen P, Schopler R: Environmental and 65. Jobbins S, Sanderson C, Alexander K: Leptospira interrogans at
physiological factors contributing to outbreaks of Cryptospo- the human–wildlife interface in northern Botswana: A newly
ridium in coquerel’s sifaka (Propithecus coquereli) at the Duke identified public health threat, Zoonoses Public Health 2013.
Lemur Center: 1999–2007, J Zoo Wildl Med 41:438–444, 2010. 66. Dietrich M, Wilkinson DA, Soarimalala V, et al: Diversification
54. Bublitz DC, Wright PC, Rasambainarivo FT, et al: Pathogenic of an emerging pathogen in a biodiversity hotspot: Leptospira in
enterobacteria in lemurs associated with anthropogenic distur- endemic small mammals of Madagascar, Mol Ecol 23:2783–2796,
bance, Am J Primatol 77:330–337, 2015. 2014.
55. Williams CV: Prosimians. In Miller RE, Fowler ME, editors: 67. Laakkonen J: Trypomastigotes and potential flea vectors of the
Fowler’s zoo and wild animal medicine (vol 8). St. Louis, MO, endemic rodents and the introduced Rattus rattus in the rainfor-
2015, Saunders, pp 291–301. ests of Madagascar, Biodivers Conserv 12:1775–1783, 2003.
56. Reynes J-M, Andriamandimby SF, Razafitrimo GM, et al: 68. Wyatt KB, Campos PF, Gilbert MT, et al: Historical mammal
Laboratory surveillance of rabies in humans, domestic animals, extinction on Christmas island (Indian Ocean) correlates with
and bats in Madagascar from 2005 to 2010, Adv Prev Med introduced infectious disease, PLoS ONE 3:2008.
2011:727821, 2011. 69. Wilkinson DA, Mélade J, Dietrich M, et al: Highly diverse
57. Pomerantz J, Rasambainarivo FT, Dollar L, et al: Prevalence of morbillivirus-related paramyxoviruses in wild fauna of the south-
antibodies to selected viruses and parasites in introduced and western Indian Ocean islands: Evidence of exchange between
endemic carnivores in western Madagascar, J Wildl Dis 2016. introduced and endemic small mammals, J Virol 88:8268–8277,
58. Alexander AB, Poirotte C, Porton IJ, et al: Gastrointestinal para- 2014.
sites of captive and free-living lemurs and domestic carnivores in 70. Doherty TS, Glen AS, Nimmo DG, et al: Invasive preda-
eastern Madagascar, J Zoo Wildl Med 47:141–149, 2016. tors and global biodiversity loss, Proc Natl Acad Sci U S A
59. Blancou J: Note sur l’infestation des lémuriens malgaches par 113:11261–11265, 2016.
Spirocerca lupi (Rudolphi, 1809), Rev Elev Med Vet Pays Trop 71. Young HS, Parker IM, Gilbert GS, et al: Introduced species,
29:127, 1976. disease ecology, and biodiversity–disease relationships, Trends
60. Junge RE, Dutton CJ, Knightly F, et al: Comparison of bio- Ecol Evol (Amst) 2016.
medical evaluation for white-fronted brown lemurs (Eulemur 72. Russell JC, Cole NC, Zuël N, et al: Introduced mammals on
fulvus albifrons) from four sites in Madagascar, J Zoo Wildl Med western Indian Ocean islands, Glob Ecol Conserv 6:132–144,
39:567–575, 2008. 2016.
SECTION 9
Infectious Diseases
44 Techniques for Vaccinating Wildlife, 299
298
44
Techniques for Vaccinating Wildlife
MARTIN GILBERT
V
accines have been widely used for over 200 years, that could be readily produced in large quantities, providing
and have been one of the most effective means of a more cost-effective approach to managing outbreaks and
controlling infectious disease in human and veteri- disrupting rabies reservoirs than alternative strategies like
nary medicine. Despite this long history, vaccines have been culling. Ongoing programs continue to strive for rabies
relatively underutilized in free-ranging wildlife, for a range elimination in more diverse wild carnivore communities
of logistical, ethical and legal reasons.1,2 This chapter will in eastern North America, where elimination from some
summarize the current understanding of vaccine use in free- species (particularly skunks [Mephitis spp.]) remains a chal-
ranging wildlife, introducing the approaches and limitations lenge.6 More limited programs have also been undertaken to
to vaccine delivery, and key considerations in their use (see control M. bovis and CSFV in wild boar (Sus scrofa), while
also Chapters 45 and 79). For the purpose of this chapter, experimental trails are being pursued to manage M. bovis,
vaccines are defined as products that are administered and B. abortus in other species (see Table 44.1).
with the intention of evoking a specific immune response Vaccines are also used as a tool to promote the conserva-
(whether humoral and/or cell-mediated) against invasive tion of endangered species, either by raising the immune
microparasites, macroparasites, or neoplasias. Although status of the threatened population itself or by preventing
vaccines are also used for regulating reproduction of disease-mediated losses of important prey species.7,8 The
wildlife populations through immunocontraception, these field use of vaccines for conservation purposes remains
techniques have been reviewed elsewhere.3 limited (see Table 44.1), although a number of novel
products are under development.9–11 In most contemporary
Motivations for Wildlife Vaccination examples, field delivery has occurred in a reactive manner,
when an outbreak is already underway,2 although vaccines
There are three main reasons for attempting the vaccination may also be used prophylactically as part of reintroduction
of wildlife, which have implications for both the design of or translocation projects.12,13 For instance, field trials are
vaccine products and the intended outcomes of vaccination now underway using a recombinant canine distemper virus
programs. Firstly, vaccines may be used to control the trans- vaccine in Hawaiian monk seals (Neomonachus schauinslandi)
mission of pathogens of public health significance, such as to manage potential outbreaks of phocine distemper virus.14
rabies, for which wildlife reservoirs may be an important In this case, epidemiologic models suggest that reactive
source of human infection. Wildlife reservoirs may also strategies may have little impact on population survival
play a role in maintaining pathogens with agro-economic due to an extended period required to achieve protective
significance due to their impacts on domestic animals, such immunity, whereas prophylactic approaches may be more
as Mycobacterium bovis, Brucella abortus, or classical swine successful.15 Recent trials have also shown some promise
fever virus. Such vaccination programs target reservoir in reducing the impact of outbreaks of Yersinia pestis on
populations in order to reduce or eliminate spillover into populations of prairie dogs (Cynomys spp.) in the United
humans or domestic animals. In these situations, priority is States using widespread prophylactic delivery of an orally
given to maximizing vaccine coverage within the reservoir, available raccoon poxvirus-based recombinant vaccine.16
evoking immunity as reliably and cost effectively as possible, Although the trial did not eliminate cases of Yersinia infec-
with the objective of reducing transmission to acceptable tion in vaccinated populations, it was associated with a
levels, or eliminating the pathogen entirely. The potential greater abundance of prairie dogs than unvaccinated control
success of reservoir-directed strategies is exemplified by the populations. The vaccine therefore could benefit both the
elimination of rabies from western Europe through the prairie dogs themselves, as well as their ecologic dependent,
distribution of oral vaccinia-based recombinant vaccines the black-footed ferret (Mustela nigripes; a species that has
targeting red foxes (Vulpes vulpes) (Table 44.1).4,5 This also been a focus of vaccination to reduce the impact of
program relied on the availability of low cost vaccine baits canine distemper virus).17
299
300 SE C T I O N 9 Infectious Diseases
TABLE
44.1 Examples of Vaccines Used to Control Infectious Disease in Wildlife Populations in the Field
Country/ Delivery
Target Species Pathogen Region Route Circumstances Citation
Mountain gorilla Gorilla beringei Measles virus Rwanda IM OUTBREAK 51,52
beringei
Black-footed ferret Mustela nigripes Canine distemper virus United States SC CONTROL 17
Island fox Urocyon littoralis Canine distemper virus United States IM, O OUTBREAK 53,54
Red wolf Canis rufus Canine parvovirus, United States IM RELEASE 13
canine distemper
virus
Hawaiian monk Neomonachus Phocine distemper virus United States IM TRIAL 14
seal schauinslandi
Reservoir species — Rabies virus Europe O CONTROL 4,5
Reservoir species — Rabies virus North America O CONTROL 6
African wild dog Lycaon pictus Rabies virus Tanzania, IM OUTBREAK, 12,55†
South Africa RELEASE
Ethiopian wolf Canis simensis Rabies virus Ethiopia IM OUTBREAK 19,56
European rabbit Oryctolagus Viral hemorrhagic Spain SC TRIAL 7
cuniculus disease virus,
myxomatosis virus
Wild boar Sus scrofa Classical swine fever Germany O OUTBREAK 57
virus
Florida puma Puma concolor coryi Feline leukemia virus United States IM OUTBREAK 58
Cheetah Acinonyx jubatus Bacillus anthracis Namibia PAR TRIAL 59
Black rhinoceros Diceros bicornis B. anthracis Namibia IM CONTROL 59
Indian one-horned Rhinoceros unicornis B. anthracis India IM OUTBREAK 60
rhinoceros
Black rhinoceros, D. bicornis, B. anthracis Zimbabwe IM OUTBREAK 22
white Ceratotherium
rhinoceros, roan simum,
antelope, kudu, Hippotragus
waterbuck, equinus,
African buffalo Tragelaphus
hippopotamus strepsiceros,
Kobus
ellipsiprymnus,
Hippopotamus
amphibius
Prairie dog spp. Cynomys spp. Yersinia pestis United States O TRIAL 8
Black-footed ferret M. nigripes Y. pestis United States SC CONTROL 17,61,62
Common brushtail Trichosurus Mycobacterium bovis New Zealand O TRIAL 63
possum vulpecula
Eurasian badger Meles meles M. bovis United IM TRIAL 64
Kingdom
Eurasian badger M. meles M. bovis Ireland O TRIAL 65,66
Wild boar S. scrofa M. bovis Spain O TRIAL 67
Bison, elk Bison bison, Cervus Brucella abortus United States IM TRIAL 68,69
elaphus
CHAPTER 44 Techniques for Vaccinating Wildlife 301
TABLE
44.1 Examples of Vaccines Used to Control Infectious Disease in Wildlife Populations in the Field—cont’d
Country/ Delivery
Target Species Pathogen Region Route Circumstances Citation
Bighorn sheep Ovis canadensis Pasteurella multocida, United States IM OUTBREAK 70†
P. trehalosi,
Mannheimia
haemolytica
White-footed Peromyscus Borrelia burgdorferi United States O TRAIL 40,41
mouse leucopus
Koala Phascolarctos Chlamydia pecorum Australia SC TRIAL 71
cinereus
Delivery route is indicated as oral (O), intramuscular injection (IM), subcutaneous injection (SC), or unspecified parenteral route (PAR). Circumstances of vaccine
use are summarized as outbreak response (OUTBREAK), long-term control (CONTROL), prophylaxis of released animals (RELEASE), or field trial (TRIAL).
Interventions reported by authors as unsuccessful are denoted by †.
In declining populations each individual becomes to be palatable for the target species.1,20 Oral delivery may
increasingly important to maintaining overall genetic diver- also be desirable for programs with a conservation objective,
sity, and strategies that maximize vaccination coverage may but the smaller numbers of animals involved raises the
be desirable, as a means of slowing the depletion of unique possibility of other delivery methods, particularly when the
alleles. However, maintaining this level of vaccine coverage species is readily observed, or where handling is feasible.
is challenging and may be cost prohibitive in conservation However, it should be recognized that vaccines that require
programs that lack an economic or public health incentive. parenteral delivery introduce additional risks associated with
But for highly threatened populations, where the objective capture, handling, and/or immobilization.1 Alternatively,
may be to minimize the probability of disease-induced vaccines have been delivered remotely via projectile systems
extinction, it may be possible to meet program goals such as darts, or bio-bullets that enable the intramuscular
through low coverage vaccination strategies that do not release of a vaccine immunogen.21,22 Vaccines have also
attempt the elimination or eradication of the pathogen. This been delivered experimentally via sprays that introduce the
is particularly applicable in the case of multi-host pathogens immunogen via the nasal or conjunctival mucosa.23,24 This
such as rabies or canine distemper virus, where abundant might have potential applications for simultaneously vac-
populations of domestic dogs or wild mesocarnivores can cinating large numbers of animals, particularly for species
act as pathogen reservoirs and perpetual sources of infection that congregate when breeding or roosting.
for threatened hosts. Eliminating infection in the reservoir
may be impractical, but low coverage strategies that aim to Considerations for Vaccine Delivery
vaccinate small numbers of a threatened population may be
sufficient to limit the spread of infection during outbreaks, Fundamentally vaccines intended for use in wildlife must
and avert extinction through the persistence of immune meet the same standards of safety and efficacy of products
survivors.18 This strategy was successful in halting the spread intended for humans or domestic species. However, the
of rabies virus during an outbreak affecting Ethiopian circumstances of delivery to free-ranging animals raise a
wolves (Canis simensis).19 An initial dose of an inactivated distinct and additional set of challenges.
vaccine (Nobivac Rabies, Intervet, Milton Keynes, United
Kingdom) was given to trapped wolves, which elicited an Safety
antibody response by 1 month post inoculation, although
a booster dose at 1–6 months was required to maintain As with any vaccine, products used in wildlife should not
antibody titers. induce disease, or lead to other deleterious side effects in
target species. This presents a challenge in the case of threat-
Methods of Delivery ened hosts where ethical or practical constraints prohibit
safety trials in target species, and it may be necessary to
The availability of vaccines that can be delivered orally draw inference from trials in a closely related model species.
is hugely beneficial for wildlife vaccination programs, Furthermore, it may also be necessary to consider vaccine
particularly those that must attain high coverage to achieve safety in nontarget hosts (including humans), particularly
the elimination of the pathogen. Oral products have been when using oral baits or other nonparenteral methods,
successfully delivered in meat (e.g., chicken heads or rabbit where there is no assurance that a vaccine product will be
legs), or via packets embedded in a flavored matrix designed delivered exclusively to the intended target species.
302 SE C T I O N 9 Infectious Diseases
can be incorporated into more cost-effective “off the shelf ” 8. Tripp DW, Rocke TE, Streich SP, et al: Apparent field safety of a
oral delivery systems, wildlife vaccines may move beyond raccoon poxvirus-vectored plague vaccine in free-ranging prairie
the theoretically possible, to the economically feasible.48 dogs (Cynomys spp.), Colorado, USA, J Wildl Dis 51:401–410,
2015.
Just as technologic advances open new opportunities,
9. Kreiss A, Brown GK, Tovar C, et al: Evidence for induction of
they may also introduce new challenges, and following the humoral and cytotoxic immune responses against devil facial
experiences of the AWD vaccination in the Serengeti, per- tumor disease cells in Tasmanian devils (Sarcophilus harrisii)
ceived or actual failure can have far-reaching consequences. immunized with killed cell preparations, Vaccine 33:3016–3025,
One conceptual development is the proposed release of 2014.
self-disseminating vaccines, such as an experimental Ebola 10. Warfield KL, Goetzmann JE, Biggins JE, et al: Vaccinating
virus vaccine intended for use in protecting great apes in captive chimpanzees to save wild chimpanzees, Proc Natl Acad
Central Africa. The proposed recombinant vaccine uses a Sci 111:1–4, 2014.
cytomegalovirus (CMV) that is assumed to be safe and 11. Tsuda Y, Parkins CJ, Caposio P, et al: A cytomegalovirus-based
host-specific, to carry Ebola virus nucleoprotein, and has vaccine provides long-lasting protection against lethal Ebola virus
been shown to protect both mice and rhesus macaques challenge after a single dose, Vaccine 33:2261–2266, 2015.
12. Hofmeyr M, Bingham J, Lane EP, et al: Rabies in African wild
from experimental challenge.11,49 While this approach could
dogs (Lycaon pictus) in the Madikwe Game Reserve, South Africa,
overcome the logistical difficulties of vaccinating rare and Vet Rec 146:50–52, 2000.
cryptic species in remote areas, it requires a high level of 13. Harrenstein LA, Munson L, Ramsay EC, et al: Antibody
confidence in the safety and host-specificity of the product, responses of red wolves to canine distemper virus and canine
as containment would be impossible once released into the parvovirus, J Wildl Dis 33:600–605, 1997.
ecosystem. However, the pivotal assumption that CMV is 14. Malakoff D: A race to vaccinate rare seals, Science 352:1265,
clinically inconsequential is debatable, as the virus is con- 2016.
sidered the main infectious cause of human birth defects in 15. Baker JD, Harting AL, Barbieri MM, et al: Modeling a morbil-
the United States (with approximately twice the incidence livirus outbreak in Hawaiian monk seals to aid in the design of
of Down’s syndrome).50 There could be considerable public mitigation programs, J Wildl Dis 53:2017.
health ramifications of releasing a recombinant and trans- 16. Rocke TE, Tripp DW, Russell RE, et al: Sylvatic plague vaccine
partially protects prairie dogs (Cynomys spp.) in field trials,
missible CMV, not to mention the untested susceptibility
Ecohealth 1–13, 2017, doi:10.1007/s10393-017-1253-x.
of our close relatives, and primary vaccine target, the great 17. US Fish and Wildlife Service: Recovery plan for the black-footed
apes. Even if safety could be ensured, the acceptance by ferret (Mustela nigripes): Second Revision, 2013.
the general public of disseminating a genetically modified 18. Haydon DT, Randall DA, Matthews L, et al: Low-coverage vac-
virus into the wild should not be assumed. Ultimately, cination strategies for the conservation of endangered species,
societal and political support are essential components of Nature 443:692–695, 2006.
successful wildlife vaccination programs and should not 19. Knobel DL, Fooks AR, Brookes SM, et al: Trapping and vaccina-
be ignored. tion of endangered Ethiopian wolves to control an outbreak of
rabies, J Appl Ecol 45:109–116, 2008.
20. Cross ML, Buddle BM, Aldwell FE: The potential of oral vac-
References cines for disease control in wildlife species, Vet J 174:472–480,
2007.
1. Blancou J, Gilot-Fromont E, Artois M, et al: Options for the 21. Olsen SC, Kreeger TJ, Schultz W: Immune responses of bison
control of disease 1: Targeting the infectious or parasitic agent. to ballistic or hand vaccination with Brucella abortus strain RB51,
In Delahay RJ, Smith GC, Hutchings MR, editors: Management J Wildl Dis 38:738–745, 2002.
of disease in wild mammals, Tokyo, Japan, 2009, Springer Japan, 22. Clegg SB, Turnbull PCB, Foggin CM, et al: Massive outbreak
pp 97–121. of anthrax in wildlife in the Malilangwe Wildlife Reserve, Zim-
2. Woodroffe R: Managing disease threats to wild mammals, Anim babwe, Vet Rec 160:113–118, 2007.
Conserv 2:185–193, 1999. 23. Corner LAL, Norton S, Buddle BM, et al: The efficacy of bacille
3. Kirkpatrick JF, Lyda RO, Frank KM: Contraceptive vaccines for Calmette-Guérin vaccine in wild brushtail possums (Trichosurus
wildlife: a review, Am J Reprod Immunol 66:40–50, 2011. vulpecula), Res Vet Sci 73:145–152, 2002.
4. Mähl P, Cliquet F, Guiot AL, et al: Twenty year experience of 24. Corner LA, Buddle BM: Conjunctival vaccination of the brush-
the oral rabies vaccine SAG2 in wildlife: a global review, Vet Res tail possum (Trichosurus vulpecula) with bacille Calmette-Guérin,
45:1–17, 2014. N Z Vet J 53:133–136, 2005.
5. Müller T, Freuling CM, Wysocki P, et al: Terrestrial rabies control 25. Blancou J: Ecology and epidemiology of fox rabies, Rev Infect Dis
in the European Union: historical achievements and challenges 10:S604–S605, 1988.
ahead, Vet J 203:10–17, 2015. 26. Fry TL, Atwood T, Dunbar M: Evaluation of rhodamine B as a
6. Slate D, Rupprecht CE, Rooney JA, et al: Status of oral rabies biomarker for raccoons, Hum-Wildl Interact 4:275–282, 2010.
vaccination in wild carnivores in the United States, Virus Res 27. Fry T, Van Dalen K, Hurley J, et al: Mucosal adjuvants to
111:68–76, 2005. improve wildlife rabies vaccination, J Wildl Dis 48:1042–1046,
7. Calvete C, Estrada R, Lucientes J, et al: Effects of vaccination 2012.
against viral haemorrhagic disease and myxomatosis on long-term 28. Edmonds MD, Samartino LE, Hoyt PG, et al: Oral vaccination
mortality rates of European wild rabbits, Vet Rec 155:388–392, of sexually mature pigs with Brucella abortus vaccine strain RB51,
2004. Am J Vet Res 62:1328–1331, 2001.
304 SE C T I O N 9 Infectious Diseases
29. Aldwell FE, Keen DL, Parlane NA, et al: Oral vaccination with 51. Hastings B, Kenny D, Lowenstine L, et al: Mountain gorillas and
Mycobacterium bovis BCG in a lipid formulation induces resis- measles - ontogeny of a wildlife vaccination program, Proc Annu
tance to pulmonary tuberculosis in brushtail possums, Vaccine Meet Am Assoc Zoo Vet 198–207, 1991.
22:70–76, 2003. 52. Spelman LH, Gilardi KV, Lukasik-Braum M, et al: Respira-
30. Burgess DC, Burgess MA, Leask J: The MMR vaccination and tory disease in mountain gorillas (Gorilla beringei beringei) in
autism controversy in United Kingdom 1998–2005: inevitable Rwanda, 1990–2010: outbreaks, clinical course, and medical
community outrage or a failure of risk communication?, Vaccine management, J Zoo Wildl Med 44:1027–1035, 2013.
24:3921–3928, 2006. 53. Timm SF, Munson L, Summers BA, et al: A suspected canine
31. Jansen VA, Stollenwerk N, Jensen HJ, et al: Measles outbreaks distemper epidemic as the cause of a catastrophic decline in
in a population with declining vaccine uptake, Science 301:804, Santa Catalina Island foxes (Urocyon littoralis catalinae), J Wildl
2003. Dis 45:333–343, 2009.
32. Burrows R: Rabies in wild dogs, Nature 359:277, 1991. 54. Vickers TW, Garcelon DK, Fritcher DL, et al: Evaluation of
33. Creel S: Cause of wild dog deaths, Nature 360:633, 1992. an oral and food-based canarypox-vectored canine distemper
34. Heinsohn R: When conservation goes to the dogs, Trends Ecol vaccine in endangered Channel Island Foxes (Urocyon Littoralis),
Evol (Amst) 7:214–215, 1992. Proc AAZV, AAWV, WDA Jt Conf: 87–88, 2004.
35. Macdonald DW: Causes of wild dog deaths, Nature 360:633–634, 55. Gacoyne SC, King AA, Laurenson MK, et al: Aspects of rabies
1992. infection and control in the conservation of the African wild dog
36. Burrows R, Hofer H, East ML: Extinction and intervention in a (Lycaon pictus) in the Serengeti region, Tanzania, Onderstepoort J
small population: the case of the Serengeti wild dogs, Proc R Soc Vet Res 60:415–420, 1993.
London B 256:281–292, 1994. 56. Randall DA, Marino J, Haydon DT, et al: An integrated disease
37. Dye C: Serengeti wild dogs: what really happened?, Trends Ecol management strategy for the control of rabies in Ethiopian
Evol (Amst) 11:188–189, 1996. wolves, Biol Conserv 131:151–162, 2006.
38. Woodroffe R: Assessing the risks of intervention: immobiliza- 57. Moennig V: The control of classical swine fever in wild boar,
tion, radio-collaring and vaccination of African wild dogs, Oryx Front Microbiol 6:1–10, 2015.
35:234–244, 2001. 58. Cunningham MW, Brown MA, Shindle DB, et al: Epizootiology
39. Cleaveland S, Mlengeya T, Kaare M, et al: The conservation and management of feline leukemia virus in the Florida puma, J
relevance of epidemiological research into carnivore viral diseases Wildl Dis 44:537–552, 2008.
in the Serengeti, Conserv Biol 21:612–622, 2007. 59. Turnbull PC, Tindall BW, Coetzee JD, et al: Vaccine-induced
40. Bhattacharya D, Bensaci M, Luker KE, et al: Development of a protection against anthrax in cheetah (Acinonyx jubatus) and
baited oral vaccine for use in reservoir-targeted strategies against black rhinoceros (Diceros bicornis), Vaccine 22:3340–3347,
Lyme disease, Vaccine 29:617–636, 2011. 2004.
41. Tsao JI, Wootton JT, Bunikis J, et al: An ecological approach 60. Pandit P, Sinha S: Anthrax incidence and its control by vac-
to preventing human infection: vaccinating wild mouse reser- cinating greater one horned rhino (Rhinoceros unicornis) against
voirs intervenes in the Lyme disease cycle, Proc Natl Acad Sci anthrax in Jaldapara Wildlife Sanctuary, West Bengal, India,
101:18159–18164, 2004. Indian For 132:45–65, 2006.
42. Chambers MA, Carter SP, Wilson GJ, et al: Vaccination against 61. Rocke TE, Smith S, Marinari P, et al: Vaccination with F1-V
tuberculosis in badgers and cattle: an overview of the challenges, fusion protein protects black-footed ferrets (Mustela nigripes)
developments and current research priorities in Great Britain, Vet against plague upon oral challenge with Yersinia pestis, J Wildl
Rec 175:90–96, 2014. Dis 44:1–7, 2008.
43. Gilbert M, Soutrina S, Seryodkin I, et al: Canine distemper virus 62. Abbott RC, Osorio JE, Bunck CM, et al: Sylvatic plague vaccine:
as a threat to wild tigers in Russia and across their range, Integr a new tool for conservation of threatened and endangered
Zool 10:329–343, 2015. species?, Ecohealth 9:243–250, 2012.
44. Sadler RA, Ramsay E, McAloose D, et al: Evaluation of two 63. Tompkins DM, Ramsey DS, Cross ML, et al: Oral vaccination
canine distemper virus vaccines in captive tigers (Panthera tigris), reduces the incidence of tuberculosis in free-living brushtail
J Am Vet Med Assoc 47:558–563, 2016. possums, Proc R Soc London B Biol Sci 276:2987–2995, 2009.
45. Cassone A: Fungal vaccines: real progress from real challenges, 64. Chambers MA, Rogers F, Delahay RJ, et al: Bacillus Calmette-
Lancet Infect Dis 8:114–124, 2008. Guérin vaccination reduces the severity and progression of
46. McMahon TA, Sears BF, Venesky MD, et al: Amphibians acquire tuberculosis in badgers, Proc Biol Sci 278:1913–1920, 2011.
resistance to live and dead fungus overcoming fungal immuno- 65. Aznar I, McGrath G, Murphy D, et al: Trial design to estimate
suppression, Nature 511:224–227, 2014. the effect of vaccination on tuberculosis incidence in badgers, Vet
47. Stice MJ, Briggs CJ: Immunization is ineffective at preventing Microbiol 151:104–111, 2011.
infection and mortality due to the amphibian chytrid fungus 66. Corner L, Gormley E: Control of TB in wildlife by oral BCG
Batrachochytrium dendrobatidis, J Wildl Dis 46:70–77, 2010. vaccination, Expert Rev Vaccines 8:1339, 2009.
48. Corro LM, Tripp DW, Stelting SA, et al: Using off-the-shelf 67. Beltrán-Beck B, Ballesteros C, Vicente J, et al: Progress in oral
technologies to mass manufacture oral vaccine baits for wildlife, vaccination against tuberculosis in its main wildlife reservoir in
J Wildl Dis 53:2017. Iberia, the Eurasian wild boar, Vet Med Int 2012:978501, 2012.
49. Marzi A, Murphy AA, Feldmann F, et al: Cytomegalovirus-based 68. Kreeger TJ, Cook WE, Edwards WH, et al: Brucella abortus
vaccine expressing Ebola virus glycoprotein protects nonhuman strain RB51 vaccination in elk. II. Failure of high dosage to
primates from Ebola virus infection, Sci Rep 6:21674, 2016. prevent abortion, J Wildl Dis 38:27–31, 2002.
50. Cheeran MC, Lokensgard JR, Schleiss MR: Neuropathogenesis 69. Olsen SC, Christie RJ, Grainger DW, et al: Immunologic
of congenital cytomegalovirus infection: disease mechanisms and responses of bison to vaccination with Brucella abortus strain
prospects for intervention, Clin Microbiol Rev 22:99–126, 2009. RB51: comparison of parenteral to ballistic delivery via
CHAPTER 44 Techniques for Vaccinating Wildlife 305
compressed pellets or photopolymerized hydrogels, Vaccine 24: 71. Waugh C, Khan SA, Carver S, et al: A prototype recombinant-
1346–1353, 2006. protein based Chlamydia pecorum vaccine results in reduced
70. Cassirer EF, Rudolph KM, Fowler P, et al: Evaluation of ewe vac- chlamydial burden and less clinical disease in free-ranging
cination as a tool for increasing bighorn lamb survival following koalas (Phascolarctos cinereus), PLoS ONE 11:e0146934,
pasteurellosis epizootics, J Wildl Dis 37:49–57, 2001. 2016.
45
Brucellosis in North American Wildlife
JACK C. RHYAN AND PAULINE NOL
Brucellosis: An Introduction The most recent discoveries of new Brucella species were
B. papionis in captive baboons (Papio spp.),8 B. vulpis in
Brucellosis is a group of diseases caused by bacterial species red foxes in Austria,9 strains closely related to B. vulpis in
in the genus Brucella. Brucella spp. organisms are small, rodents (Myodes glareolus; Microtis spp.) and shrews (Sorex
nonspore-forming gram-negative coccobacilli that live and spp.) in Germany,10 and as yet unnamed strains found in
replicate inside the host’s cells, especially phagocytic cells Australian rodents (Rattus assimilis; Melomys spp.).11
and placental trophoblasts. Classically, each species has
been subdivided into biovars dependent on the presence of
certain antigens as determined by reacting the organisms Cooperative State-Federal Brucellosis
with different antisera. Since genomic typing has developed, Eradication Program
however, the relationship between genotypes and biovars is
being reevaluated. To understand the importance of the presence of brucel-
Most, but not all, Brucella spp. cause zoonotic disease. losis in wildlife reservoirs in North America, familiarity
Brucellosis in humans is known as undulant fever and is with the 80-year eradication effort in the United States is
characterized by persistent and recurrent bouts of fever, helpful. In the early 1900s, brucellosis was considered the
chills, night sweats, headache, arthralgia, arthritis, anorexia, most economically devastating disease of livestock, and in
nausea, weight loss, malaise, and dementia. Brucella infec- 1934, the Cooperative State-Federal Brucellosis Eradication
tions in humans usually respond to antibiotic therapy, Program was begun. The program has used quarantine, test
although recrudescence is not uncommon and polymerase and slaughter, calfhood vaccination, and, more recently,
chain reaction (PCR) indicates that brucellar DNA may be adult vaccination to accomplish its goal. In 1934, nearly
demonstrated in patients years after antibiotic treatment.1 50% of the cattle herds tested had seropositive or suspect
The causative organism of a severe, sometimes fatal, animals and by the 1990s less than 200 affected herds
disease in soldiers in Malta in 1887 was identified by remained. For over a decade cattle and captive bison (Bison
David Bruce and later given the name Brucella melitensis. bison) herds, nationwide, have been negative for brucellosis
A similar organism, B. abortus, was identified a decade later with the exception of herds in the Greater Yellowstone
by Bernard Bang as causing “infectious abortion” in cattle. Area (GYA) that acquire the infection from free-ranging elk
In the following decades, B. suis was identified in domestic (Cervus canadensis).12 National and state efforts to control
swine, B. canis in dogs, B. ovis in sheep, and B. neotomae swine brucellosis were begun in the late 1950s, at which
in desert woodrats (Neotoma tepida). Until recently, those time the disease was prevalent in domestic herds. Currently,
organisms comprised the genus Brucella. However, since swine brucellosis has been eradicated from the commercial
1994, several new species of Brucella have been identified. swine industry.
In marine mammals, two new species have been reported
worldwide (currently named B. pennipedialis and B. ceti, Brucellosis in Wildlife
in seals and cetaceans respectively), and are reviewed by
Nymo et al., 2011.2 Brucella inopinata has been identified There are four general ecosystem infections of brucellosis
in numerous frog species (reviewed by Mühldorfer et al., in North American wildlife: (1) B. abortus in bison and
2016).3 Brucella microti was first isolated common voles elk in the GYA and in bison in and around Wood Buffalo
(Microtus arvalis) in the Czech Republic4 and subsequently National Park (WBNP), Canada; (2) B. suis in invasive wild
has been found to be present in the soil from the same swine; (3) B. suis biovar 4 in caribou (Rangifer tarandus)
area,5 and in addition, was later isolated in red foxes (Vulpes and other wildlife in the Arctic; and (4) B. ceti and B. pin-
vulpes) in Austria and a wild boar (Sus scrofa) in Hungary.6,7 nipedialis in marine mammals. The successful eradication
306
CHAPTER 45 Brucellosis in North American Wildlife 307
of B. abortus from cattle, ranched bison, and most public Brucellosis in Wood Buffalo National
bison herds in the United States and Canada leaves bison Park Bison
and elk in the GYA and bison in WBNP as the remaining
reservoirs of the infection. Brucella suis remains present in Canada’s sole known wildlife reservoir of brucellosis consists
many wild swine populations and occurs occasionally in of several subpopulations of bison within and adjacent to
backyard or “transitional” domestic swine operations having WBNP in Alberta and the Northwest Territories. Currently,
contact with wild swine. bison populations within the greater WBNP region are
There are many published surveys of wildlife for brucel- estimated at 4500 animals and have generally increased in
losis, although most are limited to examination of serum for recent years in spite of a coinfection with bovine tubercu-
antibodies to Brucella spp. The serologic tests used are usually losis and predation by wolves.16 In a survey conducted in
those developed for cattle to detect antibodies to B. abortus. the mid-1980s, 25% of 72 WBNP bison examined had
When using these tests to detect antibodies to other species evidence (culture and/or serology) of B. abortus infection.17
of Brucella, one is dependent on cross-reacting antibodies
developed against shared antigens in the organism’s cell wall. The Disease in Bison
Factors to take into account when interpreting the results of
these studies include: undetermined or variable specificities In bison, B. abortus is transmitted primarily through
and sensitivities of tests when used in nonbovine species; ingestion or mucosal contact with organisms shed in the
detection of antibodies indicates possible exposure to the placenta, fetus, and birth fluids at abortion or calving. It is
agent, not necessarily active infection; and false positive also shed in the milk; however, it appears that calves often
results may occur due to cross-reactive antibodies developed clear any infection and become seronegative by weaning.
on exposure to several non-Brucella bacterial species, such In fact, 5-month-old calves may be seronegative, though
as Yersinia enterocolitica strain 09. In the case of marine suckling milk from which B. abortus can be isolated.18 Post-
mammals, considerable variation on the same serum sample weaning seronegative calves, regardless of prior serostatus
between standard B. abortus tests has been observed and due to maternal antibody, are susceptible to infection and
numerous assays have been developed specifically for abortion. Seroconversion to positive rates (indicating at
detection of antibodies to brucellosis in pinnipeds and least exposure to B. abortus) in the GYA are 20% per year
cetaceans. for calves up to 3 years of age and then decrease to 10%
per year.18
Latent infection, also known as the “heifer syndrome,”
Brucellosis in Bison and Elk in the is a phenomenon which is rare but has been documented
Greater Yellowstone Area in cattle. Calves born to infected dams become latently
infected due to in utero or neonatal exposure. They become
Brucellosis was first serologically diagnosed in wild bison in seronegative after weaning and show no evidence of infec-
Yellowstone National Park in 1917, in elk in Yellowstone in tion until their first parturition in which they seroconvert
1931, and on the National Elk Refuge, Jackson, Wyoming and may abort or shed organisms. Understandably, this
in 1930. Populations of bison and elk in the GYA are phenomenon may be problematic for eradication strate-
approximately 5000 and 100,000 respectively. Brucella gies. Latent infection has not been documented in bison;
seroprevalence in GYA bison is approximately 50%–60% however, it is too early to conclude that it does not
and is highly variable in elk, ranging from negligible to 40% occur due to the limited number of observations made
depending on the year and location.13 Recent surveys in elk thus far.
show an average of 21.9% positive on feeding grounds and Among female bison exposed to B. abortus, some become
3.7% positive on land in proximity to feeding grounds.13 infected and do not abort or shed organisms; others abort
Until about 2006, it was thought that the 22 state and one or more times. Some infected cows may shed at parturi-
federal elk feeding grounds in Wyoming and intermittent tion, have one or more normal calves without shedding, and
winter feeding grounds in Idaho were necessary for main- then have an abortion with marked shedding of brucellae.
tenance and transmission of infection in elk. However, it Additionally, the infection causes metritis and mastitis in
has been observed that seroprevalence in some elk herds cows, with reduced birth rates in seropositives compared
remote from feeding grounds has markedly increased,13 to seronegatives.19 Infection of the mammary gland often
indicating expanding infection among elk populations. occurs but not as frequently as in cattle.20 Retained placenta
This has resulted in many cases of transmission to cattle may accompany abortion or calving and likely contributes
and captive bison herds in the GYA.12 Factors that may considerably to transmission among herd-mates, especially
have influenced the apparent switch from elk as a spillover juvenile male bison who are still present with the nursery
host to a maintenance host include increasing populations groups and very interested in the hanging placenta.
of elk, land use changes resulting in larger congregations of In bulls B. abortus causes seminal vesiculitis, epididy-
elk on private property during winter and spring,14 and the mitis, ampullitis, and occasionally orchitis, which may
reintroduction of wolves to the GYA which has influenced affect fertility. It is thought that venereal transmission is
elk behavior.15 not a significant factor in the epidemiology of the disease
308 SE C T I O N 9 Infectious Diseases
in bison. This is extrapolated from our knowledge of the persistent bacteremia. The disease is thought to be gener-
disease in cattle and is supported by the findings of a single ally fatal in moose, and this species is considered a likely
limited study.21 In using several seronegative bulls to breed dead-end host.
positive cows, we have not seen seroconversion in the bulls. In many areas of Alaska and Canada, moose share habitat
In a study collecting semen from wild bison bulls in the with caribou and reindeer. Brucella suis biovar 4 infects
spring (prebreeding season), 1–8 CFU B. abortus/mL of caribou and reindeer (see section “Brucellosis in Caribou”)
semen could be recovered.22 In recently infected bulls, and has been isolated from bone lesions from a wild
we have isolated up to 2000 CFU B. abortus per mL of debilitated moose killed in the Northwest Territories.27 An
semen. After instilling 10 × 107 CFU B. abortus strain 19 experimental infection of a moose with biovar 4 produced
into the vagina of cows at breeding, we observed transient severe clinical illness, and infection of blood, lymph nodes,
seroconversion but were unable to isolate the organism from liver, and spleen.28 It was the authors’ opinion the infection
tissues 6 months later.23 Nonreproductive tract lesions in would have been fatal in the wild.
bison are occasionally seen and include arthritis, abscesses,
bursitis, and hygroma formation. Following infection, Brucellosis in Bighorn Sheep
bison develop antibodies to B. abortus, detectable on several
tests currently used for cattle. The success in obtaining Brucella abortus infection has been observed in bighorn sheep
a B. abortus isolate from a seropositive bison varies with (Ovis canadensis) in a research facility and was attributed to
the stage of infection, the harvest of appropriate tissues, fence-line contact with aborting infected elk.29 The infection
and the rigor of the isolation process. In recently infected produced abortion, placentitis, orchitis, epididymitis, and
animals, a large percentage of seropositives will be culture lymphadenitis in the bighorns and apparently resulted in
positive.24 In a small sample of random seropositive bison the death of two rams. The organism has not been isolated
from the GYA, we isolated the organism from 47% of from bighorns in the wild.
seropositives.20 Antibody levels to B. abortus may slowly In North America, serological titers to B. ovis have been
decrease over time but are surprisingly persistent with only a detected in wild bighorn sheep. An experimental infec-
rare animal changing from seropositive to seronegative over tion of bighorns demonstrated seroconversion, abortion,
several years.18 epididymitis, ampullitis, seminal vesiculitis, prostatitis, and
necrotizing and pyogranulomatous orchitis (Fig. 45.1).30
The Disease in Elk Compared with domestic sheep, bighorns developed more
frequently detectable bacteremia and equivalent or more
The pathogenesis and clinical disease in elk are similar severe lesions.
to what is seen in bison, although reports vary regarding
percentage of abortions in infected elk. In male elk, epi- Brucellosis in Wild Pigs
didymides, seminal vesicles, and ampullae may be infected
with B. abortus. Also in elk, bursa, joint, and tendon sheath Brucella suis infection was first isolated in an invasive wild
infections due to B. abortus are not uncommon. Infection pig in North America in 1974 in an animal collected in
of the mammary gland is not as prevalent in elk as in cattle. South Carolina, USA.31 In the following four decades, B.
There appear to be differences in intraspecies transmissibil- suis has been isolated from wild pigs in eight additional
ity of brucellosis in elk and bison with elk being less efficient United States including Hawaii. Brucella suis biovar 1
transmitters. This is likely, at least in part, due to behavioral predominates in North America; however, biovar 3 has
differences. Calving events among bison, especially early been identified in a Hawaiian wild pig.32 In another five
in the calving season, attract other cows and calves that states there is evidence of Brucella exposure in wild pigs
intimately interact with the birthing process through sniff- in the form of antibody detection. As serologic tests are
ing and licking. In contrast, elk generally calve away from not very sensitive in detecting brucellosis in swine, it is
the herd and maintain some isolation for several days, even possible that the disease may go underreported in some
in confinement. Of course, the presence of a feeding ground areas.32,33 At least 38 states in the United States and 3
changes this isolation dynamic considerably as the infected provinces of Canada are known to harbor invasive wild pigs
cow elk may abort or dribble infectious discharge on the with numbers likely exceeding 5 million.34,35 The presence
feedline, exposing others to the agent. of B. suis in North American wild pig populations that
have great potential for continued range expansion poses a
Brucellosis in Moose constant threat to the domestic swine industry (especially
transitional swine herds) and other domestic animals, as
Natural and experimental B. abortus infection has been well as to human health. However, it is difficult to predict
reported in the moose (Alces alces).25,26 Reported lesions how the prevalence and range of brucellosis in wild pigs
include pericarditis, myocarditis, pleuritis, peritonitis, may change with changing wild pig populations, as there is
arteritis, lymphadenitis, orchitis, hepatitis, and arthritis. little information regarding the epidemiology of brucellosis
In experimentally infected moose, antibody responses in these animals in North America. Much more research
developed within 15 days and remained high along with must be done regarding the epidemiology of this disease in
CHAPTER 45 Brucellosis in North American Wildlife 309
• Figure 45.1 Bighorn sheep ewe (Ovis canadensis) experimentally infected with Brucella ovis aborting
fetus and ram inspecting the fetus indicating possible route of transmission. (Photos courtesy of Matt
McCollum.)
wild pigs in order to be able to effectively understand and organism that was classified as B. suis biovar 4.39 A survey
manage it in these free-ranging populations. conducted in the 1960s found that in communities where
Brucella suis infection in swine may produce the fol- caribou were eaten, 5%–20% of the residents had positive
lowing clinical signs: weak or stillborn piglets, abortion titers to Brucella. The infection is pan-Arctic and whereas
(although less common than in cattle infected with B. R. tarandus is the natural host, other species are susceptible
abortus), orchitis, and lameness and posterior paralysis including muskox, foxes, moose, dogs, wolves, bears, and
due to arthritis. Associated lesions may include placentitis, some rodents.28,40–42 At present, the seroprevalence of B.
seminal vesiculitis, epididymitis, arthritis, and abscessation suis biovar 4 infection is estimated to be less than 5%
of various tissues. Transmission in swine generally occurs in Alaskan reindeer (R. Gerlach, personal communication,
through the venereal route as well as by exposure to aborted December 12, 2016) and highly variable in caribou, occa-
fetuses or other products of parturition. Brucella suis infec- sionally exceeding 40% in Canada.43 Brucellosis has not
tion in humans and other species may occur through aerosol been diagnosed in imported reindeer in the lower 48 states
and oral routes, as well as via breaks in the epidermis, and of the United States or in the small number of woodland
may be severe. There are several reports of B. suis being caribou that range in southern British Columbia, northwest
contracted by slaughterhouse workers and hunters while Montana, and northern Idaho.
butchering or field dressing infected pigs.36,37 Transmission of the disease is by ingestion of or contact
with infected products of parturition. Whether venereal
Brucellosis in Caribou transmission or vertical transmission through the milk
occurs is unknown. In R. tarandus, B. suis biovar 4 produces
The northern portions of North America are home to abortion, weak calves, retained placentas, lameness, sterility,
hundreds of thousands of free-ranging caribou occupying orchitis, epididymitis, seminal vesiculitis, metritis, mastitis,
boreal, montane, and arctic environments. Additionally, nephritis, and bursitis.44 An alternate pathway of transmis-
there are approximately 20 herds of reindeer and domesti- sion was tested by Vashkevich (1978),45 who demonstrated
cated caribou, often also free-ranging and located primarily transovarian transmission of brucellosis from infected to
in the Seward Peninsula and Aleutian Islands of Alaska. naïve reindeer by the warble fly (Oedemagena tarandi).
Reindeer and their herders comprise a small but growing Disease management tools for reindeer consist of vaccina-
commercial livestock industry whereas hunting of caribou tion (adjuvented, killed B. suis biovar 4) and test and cull
provides meat and other animal products for a large number of seropositives. Currently no specific disease management
of indigenous peoples in proximity to those animals. is practiced in caribou.
Brucellosis was serologically first detected in humans in
Alaska in 1939. Subsequently, an organism resembling B. Brucellosis in Wild Carnivores
suis was isolated from human blood and bone marrow, and
epidemiologic studies suggested caribou as a source of infec- Several serologic surveys have indicated exposure of North
tion. In 1963, the organism was isolated from caribou,38 American wild carnivores to Brucella spp., summarized by
and later studies demonstrated the human and reindeer Davis (1990).46 These species include wolves, arctic foxes,
isolates from Alaska, Canada, and Siberia were the same red foxes, coyotes, and black, grizzly, and polar bears42;
310 SE C T I O N 9 Infectious Diseases
however, naturally occurring disease has not been reported and a common dolphin (Delphinus delphis) found dead
in these carnivores. Experimental infections have suggested on the Scottish coast.49 Also in 1994, a similar organism
that carnivores usually develop detectable antibodies and was isolated from an aborted fetus of a captive bottlenose
transient infection of lymph nodes and organs but do not dolphin (Tursiops truncatus) in California.50,51 Brucella spp.
usually shed significant numbers of brucellae.47 Transmis- infections have since been detected by serology and/or
sion of B. abortus from coyotes to cattle has been observed culture in numerous species of cetaceans and pinnipeds,
under experimental, close-confinement conditions,48 but with nearly global distribution.2,52 Additionally, a recent
wild carnivores are generally considered “spillover” hosts report describes the isolation of a unique Brucella strain
and are not thought to be significant transmitters of infec- from an osteolytic lesion of a southern sea otter (Enhydra
tion in nature.46 In selected areas such as the GYA, wild lutris nereis) on the coast of California.53 Molecular and
canids could serve as effective sentinels for the presence of biochemical data suggest the organism represents a novel
brucellosis in free-ranging ungulates. lineage distinct from B. ceti and B. pinnipedialis though
more closely related to pinniped strains. Reports of evidence
Brucellosis in Marine Mammals of Brucella spp. infection or exposure in North American
marine mammals are summarized in Table 45.1.
Brucella spp. were first isolated in 1994 from harbor seals Brucella ceti has been associated with a variety of lesions in
(Phoca vitulina), harbor porpoises (Phocoena phocoena), several cetacean species (see Table 45.1) including abortions
TABLE Summary of Publications Reporting Evidence of Brucella spp. Infection or Exposure in Marine Mammals
45.1 by Species
TABLE Summary of Publications Reporting Evidence of Brucella spp. Infection or Exposure in Marine Mammals
45.1 by Species—cont’d
Common Name Type of Tests
(Species) Location Pathology Performed Reference
New England, USA N/O Serology, Culture, Maratea et al.
Histopathology, (2003)58
Immunohistochemistry
Atlantic Coast, USA; St. N/A Serology Nielsen et al. (2001)67
Lawrence Estuary, Canada;
Nova Scotia, Canada;
Vancouver Island, Canada
Puget Sound, WA, USA Pulmonary abscess Serology, Culture, Lambourn et al.
Histopathology, (1998)71
Immunohistochemistry
Hooded seal St. Lawrence Estuary, Canada; N/A Serology Nielsen et al. (2001)67
(Cystophora Gulf of St. Lawrence,
cristata) Canada; Nova Scotia,
Canada; Newfoundland,
Canada
Gray seal Gulf of St. Lawrence, Canada; N/A Serology Nielsen et al. (2001)67
(Halichoerus Nova Scotia, Canada
grypus)
Harp seal (P. Magdalen Islands, Quebec, N/O Serology, Culture Forbes et al. (2000)72
groenlandica) Canada
New England, USA N/O Serology, Culture, Maratea et al.
Histopathology, (2003)58
Immunohistochemistry
Gulf of St. Lawrence, Canada; N/A Serology Nielsen et al. (2001)67
Newfoundland, Canada
Ringed seals Newfoundland, Canada; N/A Serology Nielsen et al. (2001)67
(Pusa hispida) Nunavut, Canada
Baffin Island, Nunavut, Canada N/O Serology, Culture Forbes et al. (2000)72
Nunavut, Canada; Holman N/A Serology Nielsen et al. (1996)73
Island, Northwest Territories,
Canada
Northern fur seal Alaska, USA Placentitis Serology, PCR, Duncan et al.
(Callorhinus Histopathology, (2014)56
ursinus) Immunohistochemistry
Hawaiian monk Hawaii, USA N/A Serology Nielsen et al. (2005)74
seal (Monachus
schauinslandi)
California sea San Miguel Island, CA, USA N/O Culture, Histopathology Goldstein et al.
lion (Zalophus (2011)75
californianus)
Polar bears Alaska, USA N/A Serology O’Hara et al. (2010)42
(Ursus arctos)
Atlantic walrus Nunavut, Canada N/A Serology Nielsen et al. (2001)67
(Odobenus Nunavut, Canada N/A Serology Nielsen et al. (1996)73
rosmarus
rosmarus)
Southern sea California, USA Osteolytic lesions, Culture, Histopathology, Miller et al. (2017)53
otter (Enhydra myelitis, the Immunohistochemistry
lutris) myocardial mottling,
hepatosplenomegaly
11. Tiller RV, Gee JE, Frace MA, et al: Characterization of novel Bru- 30. McCollum M, Rhyan J, Coburn S, et al: Clinical, culture,
cella strains originating from wild native rodent species in North serology, and histopathology outcomes of bighorn sheep experi-
Queensland, Australia, Appl Environ Microbiol 76:5837–5845, mentally infected with Brucella ovis, J Wildl Dis 49:900–910,
2010. 2013.
12. Rhyan JC, Nol PN, Quamce C, et al: Transmission of brucellosis 31. Wood GW, Hendricks JB, Goodman DE: Brucellosis in feral
from elk to cattle and bison, Greater Yellowstone Area, USA, swine, J Wildl Dis 12:579–582, 1976.
2002–2012, Emerg Infect Dis 19:1992, 2013. 32. Pedersen K, Quance CR, Robbe-Austerman S, et al: Identifica-
13. Scurlock BM, Edwards WH: Status of brucellosis in free-ranging tion of Brucella suis from feral swine in selected states in the USA,
elk and bison in Wyoming, J Wildl Dis 46:442–449, 2010. J Wildl Dis 50:171–179, 2014.
14. Cross PC, Cole EK, Dobson AP, et al: Probable causes of increas- 33. Stoffregen WC, Olsen SC, Wheeler CJ, et al: Diagnostic charac-
ing brucellosis in free-ranging elk of the Greater Yellowstone terization of a feral swine herd enzootically infected with Brucella,
Ecosystem, Ecol Appl 20:278–288, 2010. J Vet Diagn Invest 19:227–237, 2007.
15. Proffitt KM, White PJ, Garrott RA: Spatio-temporal overlap 34. Pimental D: Environmental and economic costs of vertebrate
between Yellowstone bison and elk – implications of wolf restora- species invasions into the United States. In Proceedings: Inter-
tion and other factors for brucellosis transmission risk, J Appl Ecol national Symposium on Managing Vertebrate Invasive Species:
47:281–289, 2010. 2007.
16. Shury TK, Nishi JS, Elkin BT, et al: Tuberculosis and brucellosis 35. Brook RK, Van Beest FM: Feral wild boar distribution and
in wood bison (Bison bison athabascae) in Northern Canada: A perceptions of risk on the central Canadian prairies, Wildl Soc
renewed need to develop options for future management, J Wildl Bull 38:486–494, 2014.
Dis 51:543–554, 2015. 36. Brucella suis infection associated with feral swine hunting – three
17. Tessaro SV, Forbes LB, Turcotte C: A survey of brucellosis and states, 2007–2008: Centers for Disease Control, 2009:618–
tuberculosis in bison in and around Wood Buffalo National Park, 621.
Canada, Can Vet J 31:174–180, 1990. 37. Trout D, Gomez TM, Bernard B, et al: Outbreak of brucellosis
18. Rhyan JC, Aune K, Roffe T, et al: Pathogenesis and epidemiology at a United States pork packing plant, J Occup Environ Med
of brucellosis in Yellowstone bison: Serologic and culture results 37:697–703, 1995.
from adult females and their progeny, J Wildl Dis 45:729–739, 38. Huntley BE, Philip RN, Maynard JE: Survey of brucellosis in
2009. Alaska, J Infect Dis 112:100–106, 1963.
19. Fuller JA, Garrot RA, White PJ, et al: Reproduction and survival 39. Meyer ME: Identification and virulence studies of Brucella
of Yellowstone bison, J Wildl Manage 71:2365–2372, 2007. strains isolated from Eskimos and reindeer in Alaska, Canada,
20. Rhyan JC, Gidlewski T, Roffe TJ, et al: Pathology of brucellosis in and Russia, Am J Vet Res 27:353–358, 1966.
bison from Yellowstone National Park, J Wildl Dis 37:101–109, 40. Forbes LB: Isolates of Brucella suis biovar 4 from animals
2001. and humans in Canada, 1982–1990, Can Vet J 32:686–688,
21. Robison CD, Davis DS, Templeton JW, et al: Conservation of 1991.
germ plasm from bison infected with Brucella abortus, J Wildl 41. Forbes LB, Tessaro SV: Transmission of brucellosis from reindeer
Dis 34:582–589, 1998. to cattle, J Am Vet Med Assoc 203:289–294, 1993.
22. Frey RK, Clarke PR, McCollum MP, et al: Evaluation of bison 42. O’Hara TM, Holcomb D, Elzer P, et al: Brucella species survey
(Bison bison) semen from Yellowstone National Park, Montana, in polar bears (Ursus maritimus) of northern Alaska, J Wildl Dis
USA, bulls for Brucella abortus shedding, J Wildl Dis 49:714–717, 46:687–694, 2010.
2013. 43. Curry PS, Elkin BT, Campbell M, et al: Filter-paper blood
23. Uhrig SR, Nol P, McCollum M, et al: Evaluation of transmission samples for ELISA detection of Brucella antibodies in caribou, J
of Brucella abortus strain 19 in bison by intravaginal, intrauterine, Wildl Dis 47:12–20, 2011.
and intraconjunctival inoculation, J Wildl Dis 49:522–526, 44. Tessaro SV, Forbes LB: Brucella suis biotype 4: a case of granulo-
2013. matous nephritis in a barren ground caribou (Rangifer tarandus
24. Rhyan JC, Holland SD, Gidlewski T, et al: Seminal vesiculi- groenlandicus L.) with a review of the distribution of rangiferine
tis and orchitis caused by Brucella abortus biovar 1 in young brucellosis in Canada, J Wildl Dis 22:479–483, 1986.
bison bulls from South Dakota, J Vet Diagn Invest 9:368–374, 45. Vashkevich RB: The reindeer warble fly (Oedemagena tarandi L.)
1997. as a vector for Brucella, Vopr Prirodnoaei Ochagovosti Bolezneaei
25. Forbes LB, Tessaro SV, Lees W: Experimental studies on 9:119–131, 1978.
Brucella abortus in moose (Alces alces), J Wildl Dis 32:94–104, 46. Davis D: Brucellosis in wildlife. In Duncan K, Na JR, editors:
1996. Animal brucellosis, Boca Raton, FL, 1990, CRC Press.
26. Corner AH, Connell R: Brucellosis in bison, elk, and moose in 47. Tessaro SV, Forbes LB: Experimental Brucella abortus infection
Elk Island National Park, Alberta, Canada, Can J Comp Med Vet in wolves, J Wildl Dis 40:60–65, 2004.
Sci 22:9–21, 1958. 48. Davis DS, Heck FC, Williams JD, et al: Interspecific transmis-
27. Honour S, Hickling KMH: Naturally occurring Brucella suis sion of Brucella abortus from experimentally infected coyotes
biovar 4 infection in a moose (Alces alces), J Wildl Dis 29:596–598, (Canis latrans) to parturient cattle, J Wildl Dis 24:533–537,
1993. 1988.
28. Dieterich RA, Morton JK, Zamke RL: Experimental Brucella 49. Ross HM, Jahans KL, MacMillan AP, et al: Brucella species
suis biovar 4 infection in a moose, J Wildl Dis 27:470–472, infection in North Sea seal and cetacean populations, Vet Rec
1991. 138:647–648, 1996.
29. Kreeger TJ, Cook WE, Edwards WH, et al: Brucellosis in captive 50. Miller WG, Adams LG, Ficht TA, et al: Brucella-induced abor-
Rocky Mountain bighorn sheep (Ovis canadensis) caused by tions and infection in bottlenose dolphins (Tursiops truncatus), J
Brucella abortus biovar 4, J Wildl Dis 40:311–315, 2004. Zoo Wildl Med 30:100–110, 1999.
314 SE C T I O N 9 Infectious Diseases
51. Ewalt D, Payeur JP, Martin BM, et al: Characteristics of a 64. Goertz CEC, Salvatore F, Bohach GA, et al: Brucella sp. verte-
Brucella species from a bottlenose dolphin (Tursiops truncatus), J bral osteomyelitis with intercurrent fatal Staphylococcus aureus
Vet Diagn Invest 6:448–452, 1994. toxigenic enteritis in a bottlenose dolphin (Tursiops truncatus), J
52. Hernández-Mora G, Palacios-Alfaro JD, González-Barrientes R: Vet Diagn Invest 23:845–851, 2011.
Wildlife reservoirs of brucellosis: Brucella in aquatic environ- 65. Niemanis AS, Koopman HN, Westgate AJ, et al: Evidence of
ments, Rev - Off Int Epizoot 32:89–103, 2013. exposure to Brucella sp. in harbor porpoises (Phocoena phocoena)
53. Miller MA, Burgess TL, Dodd EM, et al: Isolation and charac- from the Bay of Fundy, Canada, J Wildl Dis 44:480–485,
terization of a novel marine Brucella from a southern sea otter 2008.
(Enhydra lutris nereis), California, USA, J Wildl Dis 2017. 66. West KL, Levine G, Jacob J, et al: Coinfection and vertical
54. Guzmán-Verri C, González-Barrientos R, Hernández-Mora G, transmission of Brucella and morbillivirus in a neonatal sperm
et al: Brucella ceti and brucellosis in cetaceans, Front Cell Infect whale (Physeter macrocephalus) in Hawaii, USA, J Wildl Dis
Microbiol 2:3, 2012. 51:227–232, 2015.
55. Colegrove KM, Venn-Watson S, Litz J, et al: Fetal distress and in 67. Nielsen O, Stewart REA, Nielsen K, et al: Serologic survey of
utero pneumonia in perinatal dolphins during the northern Gulf Brucella spp. antibodies in some marine mammals of North
of Mexico unusual mortality event, Dis Aquat Organ 119:1–16, America, J Wildl Dis 37:89–100, 2001.
2016. 68. Hueffer K, Gende SM, O’Hara TM: Assay dependence of
56. Duncan CG, Tiller R, Mathis D, et al: Brucella placentitis and Brucella antibody prevalence in a declining Alaskan harbor seal
seroprevalence in northern fur seals (Callorhinus ursinus) of the (Phoca vitulina) population, Acta Vet Scand 55:2, 2013.
Pribilof Islands, Alaska, J Vet Diagn Invest 4:507–512, 2014. 69. Lambourn DM, Garner M, Ewalt D, et al: Brucella pinnipedialis
57. Garner MM, Lambourn DM, Jeffries SJ, et al: Evidence of Bru- infections in Pacific harbor seals (Phoca vitulina richardsi) from
cella infection in Parafilaroides lungworms in a Pacific harbor seal Washington State, USA, J Wildl Dis 49:802–815, 2013.
(Phoca vitulina richardsi), J Vet Diagn Invest 9:298–303, 1997. 70. Zarnke RL, Saliki JT, Macmillan AP, et al: Serologic survey for
58. Maratea J, Ewalt DR, Frasca S, et al: Evidence of Brucella sp. Brucella spp., phocid herpesvirus-1, phocid herpesvirus-2, and
infection in marine mammals stranded along the coast of south- phocine distempter virus in harbor seals from Alaska, 1976–1999,
ern New England, J Zoo Wildl Med 34:256–261, 2003. J Wildl Dis 42:290–300, 2006.
59. Dawson CE, Perrett LL, Stubberfield EJ, et al: Isolation and 71. Lambourn D, Jeffries S, Polzin L, et al: Disease screening of
characterization of Brucella from the lungworms of a harbor harbor seals (Phoca vitulina) from Gertrude Island, Washington.
porpoise (Phocoena Phocoena), J Wildl Dis 44:237–246, 2008. Proceedings: Puget Sound Research Conference 1998;881–889.
60. Brew SD, Perrett LL, Stack JA, et al: Human exposure to Brucella 72. Forbes L, Nielsen O, Measures L, et al: Brucellosis in ringed seals
recovered from a sea mammal, Vet Rec 144:4823, 1999. and harp seals from Canada, J Wildl Dis 36:595–598, 2000.
61. Annette HS, Will SP, Carol AG, et al: Human neurobrucel- 73. Nielsen O, Nielsen K, Stewart REA: Serologic evidence of
losis with intracerebral granuloma caused by a marine mammal Brucella spp. exposure in Atlantic walruses (Odobenus rosmarus
Brucella spp, Emerg Infect Dis 9:485, 2003. romarus) and ringed seals (Phoca hispida) of Arctic Canada, Arctic
62. McDonald WL, Jamaludin R, Mackereth G, et al: Characteriza- 49:383–386, 1996.
tion of a Brucella sp. strain as a marine-mammal type despite 74. Nielsen O, Nielsen K, Braun R, et al: A comparison of four
isolation from a patient with spinal osteomyelitis in New serologic assays in screening for Brucella exposure in Hawaiian
Zealand, J Clin Microbiol 44:4363–4370, 2006. monk seals, J Wildl Dis 41:126–133, 2005.
63. Cassle SE, Jensen ED, Smith CR, et al: Diagnosis and successful 75. Goldstein T, Gill VA, Tuomi P, et al: Assessment of clinical
treatment of a lung abscess associated with Brucella species infec- pathology and pathogen exposure in sea otters (Enhydra lutris)
tion in a bottlenose dolphin (Tursiops truncatus), J Zoo Wildl Med bordering the threatened population in Alaska, J Wildl Dis
44:495–499, 2013. 47:579–592, 2011.
46
Update on Melioidosis in Zoo
and Wild Animals
PAOLO R. MARTELLI AND HUI SUK-WAI
M
elioidosis is a severe, often fatal, systemic disease method for isolation of B. pseudomallei from soil may be
of humans and other animals caused by a gram- downloaded from www.melioidosis.info. Studies comparing
negative bacillus, Burkholderia pseudomallei. A culture and polymerase chain reaction (PCR) to screen for
wide range of animal species affected by melioidosis differs B. pseudomallei in a soil sample found a higher positivity
in their susceptibility to the infection and to the disease. rate by PCR than by culture.12,13 PCR detects DNA signal
Rising awareness of the disease resulted in an increase in from both live and dead bacteria. DNA from dead cells
melioidosis case reports from more than 20 countries. The may be excluded by using selective nucleic acid intercalating
list of species at risk continues to expand.1,2 Literature dyes, ethidium monoazide (EMA), or propidium monoa-
reports of melioidosis in zoo species represent only a fraction zide (PMA).14
of the information exchanged between zoo veterinarians Strain typing of B. pseudomallei by multilocus sequence
and managers. Imported animals in apparent good health typing (MLST) requires pure bacterial isolates.15,16 An
have spread the disease to areas previously not affected, updated library of B. pseudomallei MLST may be found
where they have been responsible for sporadic cases outside at http://bpseudomallei.mlst.net. MLST analysis of
endemic areas. An outbreak of melioidosis at the Jardin des 146 isolates from Ocean Park between 1999 and 2010,
Plantes in Paris in the 1970s was traced to either imported including 111 environmental and 35 clinical samples,
horses from Iran or a giant panda (Ailuropoda melanoleuca) demonstrated that five of the eight environmental
from China. The disease spread widely outside the zoo, strains have caused disease in animals at Ocean Park:
infecting animals and humans.3,4 This chapter will provide ST70, ST32, ST37, ST660, and ST684. The most
an update on progress made at the Hong Kong Ocean Park prevalent strain in both environmental and clinical samples
on diagnosis and treatment of infection in marine mammals is ST70.17
and birds and summarize the status of melioidosis in other Infection typically occurs after exposure to soil or water
zoo species. contaminated with B. pseudomallei. Modes of infection
include inhalation, ingestion, or through open wounds,
Etiology and Epidemiology although a definite route of infection is seldom identified.
Transplacental infection was reported in goats and pigs.1,18
B. pseudomallei is a gram-negative, bipolar staining, aerobic, Other routes such as biting insects, sexual transmission,
motile, rod-shaped bacterium. It is an environmental or suckling have been suggested but are considered very
saprophyte commonly isolated from contaminated soil, rare.19–23
surface water, and heavy rain water. B. pseudomallei is Cases of melioidosis at Ocean Park correlate with
remarkably resistant in the environment. The bacterium meteorologic conditions, with increased incidence associ-
was still cultivable after 16 years in distilled water,5 70 days ated with heavy rain and strong winds. Studies in Taiwan,
in an acid environment at pH 4.5, and 35 days in saltwater Hong Kong, and Singapore have also linked increased rates
at 32 ppt.6,7 B. pseudomallei may also live within amoeba of infection to high rainfall.24–26 In Hong Kong Ocean Park
(Acanthamoeba sp), fungi (Gigaspora sp), and plants.8–10 B. between the months of May and October, 10%–45% of
pseudomallei is sensitive to exposure to ultraviolet (UV) rainwater samples collected during heavy rainfall are posi-
light and is killed in less than 5 minutes in pools with free tive for B. pseudomallei. Other local conditions, including
chlorine concentrations higher than 0.3 ppm or oxidation- soil composition and wind speed, also play a major role in
reduction potential (ORP) readings higher than 500 mV the incidence of the disease.
(Martelli & Hui, unpublished).10a In humans, factors predisposing to melioidosis include
Recommended methods for environmental sampling are debilitating conditions such as diabetes, thalassemia, renal
published.11 A standard operating procedure on a simplified disease, alcoholism, and prolonged steroid therapy.27 In
315
316 SE C T I O N 9 Infectious Diseases
animals, decreased host immunity from illness, chronic Clinical Presentation, Pathology,
stress, steroid therapy, and the nature of the environmental
exposure are believed to be predisposing factors.1,27 Treatment, and Management
Cetaceans
Diagnosis
The clinical and pathologic presentations of melioidosis in
Positive bacterial culture of B. pseudomallei confirms the cetaceans at Ocean Park are detailed in Kinoshita.27 In the
diagnosis. Culture is 100% specific, but sensitivity may be decade since that report, there have been three additional
as low as 60%, depending on the method of sample col- cases of cetacean melioidosis, including two confirmed by
lection, media used, and expertise of the microbiologist.28 blood culture and another by serology. All cases occurred
Ashdown medium is the most reliable selective culture between May and October and only days after a typhoon
medium for the specific isolation of B. pseudomallei, although or heavy rain. All were successfully treated. Initial clinical
B. pseudomallei does grow in commercially available blood presentation was invariably acute to peracute. Early clinical
culture media. Colonies of motile, oxidase positive, gram- signs are nonspecific, such as pyrexia, lethargy, inappetence,
negative bacilli, resistant to gentamicin and colistin, sensi- and lack of response to the trainers. Pyrexia is a reliable
tive to amoxicillin-clavulanic acid, grown under aerobic early indicator of infection. Rectal temperature is measured
conditions, with a characteristic colonial morphology of at a depth of 15–20 cm. During the times of higher risk
metallic sheen and progression to dry and wrinkled colonies (wet season, typhoon season), it is advisable to take rectal
should be presumptively identified as B. pseudomallei.29 It temperature twice daily (BID). A temperature increase of
is important to collect blood for culture before antibiotic half a degree or higher is reported immediately and warrants
therapy begins because blood cultures from confirmed cases investigating further. Blood must be immediately collected
are usually negative after antibiotic treatment has begun. for analysis and culture when the previously mentioned
The API 20NE & Vitek systems (bioMerieux, France) are nonspecific symptoms are detected in a cetacean during
widely used to confirm the isolates biochemically. Matrix- the wet season or shortly after a heavy rainfall in an area
assisted laser desorption/ionization time-of-flight mass with a high incidence of melioidosis. When melioidosis is
spectrometry (MALDI-TOF MS) improves accuracy and suspected at presentation, ceftazidime at 30 mg/kg is admin-
speed.30–32 istered intravenously immediately after blood collection. All
Monoclonal antibody against exopolysaccharides on the confirmed cases of melioidosis had initial blood pictures
cell surface of B. pseudomallei, such as latex agglutination showing leukocytosis, neutrophilia, elevated fibrinogen,
(LA) and immunofluorescence assay (IFA), are highly spe- prolonged erythrocyte sedimentation rate (ESR), and very
cific and sensitive and may be used on cultures to provide low iron. Pronounced hypoferemia with values lower than
an early diagnosis.27,33,34 Lateral flow immunoassay (LFI) 10 µmol/L and as low as 2 µmol/L are the result of B.
for detection of capsular polysaccharide (CPS) antigen may pseudomallei’s secretion of malleobactin, a molecule with a
also be applied to blood, pus, sputum, urine, and bacterial superior ability to sequester iron from circulating blood.40,41
colonies.35,36 Other biochemistry changes reflect multiple organ insult
A variety of PCR detection methods for B. pseudomallei due to disseminated septicemia. Fibrinogen, measured on
have been developed. Targeted genes include type III secre- citrated blood, is a very responsive and early marker of
tion system, 16S rRNA, 23S rRNA, flagellin C, ribosomal inflammation in cetaceans. Later in the disease, hyperfer-
protein subunit, groEL, Tat-domain protein, etc.29,37 remia may manifest as a result of mixed iatrogenic and
PCR has been shown to be specific and sensitive when bacterial hepatic insult.
applied to bacterial isolates. However, PCR may have low The current treatment of the initial acute phase is inten-
specificity and sensitivity when applied directly to clinical sive and differs from the previously reported treatment only
specimens. in that quinolones are no longer used.27 The initial therapy
Serology is an essential tool in the diagnosis of individual consists of intravenous (IV) injections of ceftazidime at
infections and for mass surveillance. Indirect hemagglutina- 30 mg/kg 3 times daily (TID) for a minimum of 3 weeks, or
tion assay (IHA) is the earliest described serologic test for longer if episodes of pyrexia, leukocytosis, or hypoferremia
melioidosis and is routinely performed in endemic areas.38 persist. Seroconversion of IgG may require up to 18 days in
An updated standard operating procedure for IHA may be dolphins; hence the 3-week duration of the initial therapy.
downloaded from www.melioidosis.info. Should ceftazidime be poorly tolerated or contraindicated,
A cetacean-specific enzyme-linked immunosorbent assay meropenem at 20 mg/kg IV TID for 3 weeks could be an
(ELISA) was described by Chow (2004).39 Despite its high alternative. However, this remains theoretical and to date
specificity and sensitivity, the usefulness of the cetacean we have not encountered bacterial resistance or dolphin
ELISA is limited due to the long interval between infection intolerance to ceftazidime. Phlebitis is very likely to occur
and seroconversion that may take up to 18 days.27 after administration of IV meropenem.27
For other species, including birds, Ocean Park Hong This initial intensive therapy is followed by 20 weeks of
Kong uses a microagglutination test (MAT) developed in- eradication therapy consisting of TID oral administrations
house from multiple strains (Chan, unpublished). (PO) of amoxicillin-clavulanic acid (Clavulox) at 16 mg/kg,
CHAPTER 46 Update on Melioidosis in Zoo and Wild Animals 317
corresponding to 12.8 mg/kg of amoxicillin and 3.2 mg/kg haptoglobin. In pinnipeds, haptoglobin levels are a more
of clavulanic acid. A recent long-acting antibiotic cefovecin reliable indicator of early inflammation than fibrinogen.43
(Convenia) was found to be effective in vitro against all the Since Kinoshita’s previous report,27 an additional five cases
strains of B. pseudomallei, clinical or environmental, isolated of melioidosis were managed in three California sea lions
in Hong Kong Ocean Park (Martelli & Hui, unpublished). (Zalophus californiae) and two harbor seals (Phoca vitulina).
Dosing interval for cefovecin at 8 mg/kg subcutaneous (SC) Both harbor seals and two Californian sea lions survived.
is 17 days.42 Currently cefovecin is not being considered to The Californian sea lion that did not survive died within
treat the acute phase, but SC injections every 17 days offer hours of presentation, shortly after blood sampling and the
a tempting alternative to TID PO amoxicillin-clavulanic first injection of meropenem. The animal was extremely
acid for the eradication phase. More studies are needed (see depressed and pyrexic on presentation. B. pseudomallei was
also pinnipeds later). detected by IFA on the blood sample and cultured from all
Positive response to the treatment is characterized organs at necropsy. This was an extraordinarily aggressive
clinically by a rapid return to normal body temperature form of melioidosis.
and a gradual improvement of appetite and demeanor. In The treatment previously described in pinnipeds con-
cetaceans, daily blood samples to monitor hematology, sisted of SQ injections of meropenem 18.5 mg/kg TID for
biochemistry, and fibrinogen are used to assess efficacy of 2–3 weeks followed by an eradication therapy comprising
the treatment. Serology using ELISA is performed once of four antibiotics.27 This treatment protocol, although suc-
a week on the daily serum collected that week to capture cessful in seven of 12 cases, caused many iatrogenic compli-
the moment of seroconversion. Serology will eventually cations such as necrotizing dermatitis, iatrogenic myositis,
confirm or refute whether an animal with negative blood SQ cellulitis, hematuria, trauma due to restraint, and loss
cultures had contracted melioidosis. Animals that do not of appetite. Access to a pool had to be restricted to allow
seroconvert after 3 weeks and remain unwell require further repeat captures, something much disliked by the animal and
work-up. their handlers. Relapses during eradication treatment were
In two cases the initial leukocytosis was followed by a not rare and proved fatal in a grey seal (Halichoerus grypus).
severe leukopenia and neutropenia. A single intramuscular The current revised protocol’s most significant addition is
(IM) injection of granulocyte colony-stimulating factor the placement under general anesthesia of a central venous
(G-CSF, Neupogen) at 2 µg/kg led to a normalization of access catheter immediately after the decision to treat for
the leukogram. melioidosis is made. Through a central line, IV medications,
Supportive treatment includes 2–4 L of water per including rehydration fluids, may be easily administered
100 kg of body weight PO daily and ad hoc nutritional painlessly under voluntary behavior or with only cage
support. Liver supplements including silymarin (Legalon), restraint. The animal does not need to be confined on land
s-adenosylmethionine (SAM-e), and multivitamins are for the duration of treatment, which markedly reduces stress
given for the entire duration of the treatment including and favors recovery in aquatic species. A central line allows
the eradication phase. ad libitum blood draws in species for which blood sampling
Three strains of B. pseudomallei, ST32, ST37, and ST70, is challenging. This in turn allows a better assessment of the
were confirmed to have infected the dolphins in Hong efficacy of the treatment. Central venous catheters may be
Kong. Infections by ST37 have a better prognosis, with up left in place for months and require minimum care.44 With
to 60% survival compared with zero survival for ST32 and a central venous access, the antibiotic of choice is ceftazi-
ST70 (Martelli & Hui, unpublished). dime at 30 mg/kg IV TID for 2–4 weeks. The catheter
We believe the next milestone in the treatment of ceta- is left in place during the first months of the eradication
cean melioidosis will be the development of a permanent therapy, allowing blood draws or supplemental rehydration
venous access port to avoid repeat injections in the small and immediate IV access in case of relapse.
tail vessels. For the eradication therapy the four-drug protocol men-
tioned previously was abandoned and replaced successfully
Pinnipeds with PO amoxicillin-clavulanic acid at 16 mg/kg TID for
20 weeks. Alternatively, SQ injections of cefovecin at 4 mg/
Clinical presentations of melioidosis in pinnipeds are mostly kg once monthly for 5 months have also achieved successful
acute or peracute. Clinical signs are anorexia, lethargy and eradication.44
prostration, lack of response to trainers, and pyrexia. In In cases for which placing a central line is not possible,
facilities with high endemicity, unresponsive pinnipeds meropenem SQ at 18–20 mg/kg TID for 2–4 weeks is
should be restrained for clinical examination that includes followed by the eradication therapy described previously.
rectal temperature and blood sampling for hematology, Contrast computed tomography is advised near the end
biochemistry, and bacterial culture. As a precaution, a suit- of the treatment or in recalcitrant cases to detect residual
able antibiotic such as meropenem at 18–20 mg/kg may abscesses.
be administered SQ during the clinical examination. In Atypical clinical presentations in pinnipeds include a
confirmed cases of melioidosis, blood picture shows leuko- mammary infection that progressed to a fatal septicemia in a
cytosis, neutrophilia, marked hypoferremia, and increased California sea lion27 and a pustular alopecia in a harbor seal.
318 SE C T I O N 9 Infectious Diseases
That adult male seal presented with oscillating pyrexia, capri- ocular disease.51 Multiple small and large creamy abscesses
cious appetite, extensive pustular dermatitis, and alopecia. It are disseminated throughout the body at necropsy. Felidae,
was diagnosed with bacterial dermatitis related to molting. canidae, ursidae, and herpestidae are susceptible.1,2 Meerkats
Treatment with two different courses of antibiotics and are reportedly very susceptible to melioidosis in Thailand and
topical agents was unsuccessful. Melioidosis was diagnosed in Northern Australia47,52 yet have never become infected in
3 weeks after initial presentation based on rising antibodies Singapore (Oh, personal communication) because of differ-
(ELISA). This seal was treated with SQ cefovecin at 4 mg/ ent environmental conditions. Treatment is adopted from
kg once a month for 6 months. All symptoms regressed, primates or pinnipeds protocols but with species-specific
and recovery was complete. This was an uncharacteristic challenges. Prolonged treatments with cefovecin at 8 mg/kg
presentation of melioidosis in a marine mammal. twice monthly SQ and or trimethoprim/sulfamethozazole
In harbor seals, 4/4 infections with ST37 resulted in 25 mg/kg PO BID or amoxicillin-clavulanic 16 mg/kg PO
death. Also in harbor seals, 3/3 infections with ST70 BID may not treat septicemic states but could be attempted
survived, whereas 2/2 sea lions infected with ST70 died. in chronic cases.
Pinnipeds have also become infected with ST660, a strain
never isolated from cetaceans kept at the same park. The Herbivores
data on strains are presently too scarce for analysis.
Herbivores are regularly affected by melioidosis in endemic
Primates countries, presumably because their feeding habits favor
soil ingestion and because of the frequency of hoof and
Melioidosis is virtually undefinable in humans.40 Symptoms interdigital disease in this taxon. The list of published
in humans range from benign skin infections to fulminant species at risk is long and incomplete.1,2,47 It is prudent to
septicemia, wasting disease, and quiescent infections, assume that all herbivores and all marsupials are susceptible
although it is commonly thought of as an acute respiratory to melioidosis. Epidemics have occurred in ruminants and
infection.40,45 Symptoms of melioidosis in nonhuman pri- manifest as herds with widespread lameness, limb swelling,
mates range from septicemia, to wasting or chronic wounds. wasting disease, and lack of stamina.1 On necropsy, lung,
Multiple organ infections with disseminated abscesses are liver, and spleen abscesses and bacterial cultures will confirm
the common necropsy findings.1,2 Differential diagnosis the diagnosis. Equidae may be infected and seroconvert
must include tuberculosis and chromobacteriosis. but are overall more resistant, although melioidosis may
Great and lesser apes and New and Old World monkeys be fatal.1
have contracted melioidosis.2,46,47 Four of four Western Treatment of individual herbivores is rarely attempted
lowland gorillas (Gorilla gorilla gorilla) succumbed to and could follow any of the treatments previously men-
melioidosis shortly after their arrival in Singapore in 1983.48 tioned. The pasture or exhibit is generally the reservoir of
Two other gorillas were imported to Singapore in 1992. the pathogen and ideally would be abandoned or converted.
One died of melioidosis 6 months after arrival, and the Application of calcium oxide,53 good foot care, elevated or
remaining animal was sent to a European zoo. Respiratory hanging food troughs, sound social composition, and good
distress was the more evident clinical sign. B. pseudomallei soil drainage may lower exposure to B. pseudomallei and
was cultured from multiple organs on postmortem. It is reduce the incidence of the disease.
interesting to note that gorillas kept in Jakarta (Indonesia)
since 2002 have to date not contracted melioidosis (Oh, Birds
personal communication). This emphasizes the role of local
conditions in the occurrence of melioidosis. Strains reported The list of avian species affected by B. pseudomallei is
in primates include ST296, ST612, ST132, ST36, ST 131, growing and includes macaroni penguins (Eudyptes chryso-
and ST109.47 The human treatments for melioidosis are lophus),54 Galliformes, ratites, raptors and many psittacines,
suitable for primates. An initial IV intensive phase of 2 weeks and Columbiformes.55 Pathology includes splenic and
or longer with ceftazidime 30 mg/kg IV TID or IV merope- hepatic abscess, pneumonia, and generalized congestion.
nem 18.5 mg/kg TID followed by an eradication phase of The clinical presentation is variable, and birds are con-
12–20 weeks with PO trimethoprim/sulfamethozazole at sidered quite resistant to the disease.1,55 In endemic areas,
25 mg/kg BID or PO amoxicillin-clavulanic acid at 16 mg/ differential diagnosis of nonspecific clinical findings such
kg TID.49,50 Long-term sedation may assist with the IV as choanal congestion, radiographic evidence of pulmonary
treatment phase. Discouraging access to the contaminated congestion, splenomegaly, and hepatomegaly must include
soil or water through husbandry and enclosure design helps melioidosis. Avian patients may present with only minor or
to minimize contact with the pathogen. vague complaints such as lameness or ill-doing. Twelve cases
of avian melioidosis were diagnosed in Ocean Park between
Carnivores 1998 and 2007. Of those, 10 were presented dead (83%),
one died under treatment, and only one recovered fully
Melioidosis in carnivores may present as septicemia or as (8%). Psittacines represented 10 of the 12 cases, indicating
localized infections of the skin, limbs, or epididymis or even either a higher susceptibility of psittacines to the disease
CHAPTER 46 Update on Melioidosis in Zoo and Wild Animals 319
or, more likely, a more attentive husbandry staff for that to date. Zoo managers in endemic areas must seek access
group of birds. to excellent veterinary resources and build awareness of
In 2008 we undertook efforts to improve detection and melioidosis throughout their company and the public.
treatment of avian melioidosis. Between 2008 and 2016, In Ocean Park Hong Kong, across-the-board institu-
nine cases of avian melioidosis were diagnosed. Three tional awareness resulted in investments that reduced
were presented dead (33%); two failed to respond and contact with the pathogen and increased diagnostic and
were euthanized. Four birds recovered fully (44%), a net therapeutic capabilities. Reducing the interval between
improvement over the previous decade. The hemogram case presentation, diagnosis, and treatment of melioidosis
of avian melioidosis shows pronounced leukocytosis and markedly improved prognosis and survivability. Previously
heterophilia often more than 70,000 or even 90,000 white melioidosis accounted for a third of all marine mammal
blood cells (WBCs)/µL and 70% or higher respectively. deaths at Ocean Park,25 but there has been zero mortality
Affected psittacines demonstrate elevated aspartate trans- due to melioidosis in cetaceans since 2006. More intensive
aminase (AST) and creatine kinase (CK) accompanied management, combining behavioral training and advanced
by initial leukocytosis, heterophilia, and positive serol- vascular access, contributed to achieving zero mortality in
ogy. These enzymes decrease during treatment in spite of pinnipeds since 2008. There has also been progress in avian
repeated muscle trauma from ceftazidime IM injections. melioidosis diagnosis and treatments.
This suggests, but in no way affirms, that elevated WBCs,
heterophils, CK, and AST support a suspicion of infection Acknowledgments
by B. pseudomallei in psittacines. Paired serology using MAT
confirms an active infection and guides the treatment plan. We thank Karthiyani Krishnasamy, Vanaporn Wuthiekanun,
The current treatment for systemic avian melioidosis con- and Lee Foo Khong for valuable comments on the
sists of an intensive therapy of 2–4 weeks of IM ceftazidime manuscript.
at 100–150 mg/kg TID. The eradication phase consists of
170 mg/kg of amoxycillin-clavulanic acid PO BID for 20 References
weeks. Concurrently 20 mg/kg of terbinafine PO once daily
(SID) are administered on alternating weeks for 20 weeks 1. Rush CM, Thomas AD: Melioidosis in animals. In Ketheesan
to prevent secondary fungal disease from prolonged use of N, editor: Melioidosis: a century of observation and research,
antibiotics. Amsterdam, 2012, Elsevier, pp 313–336.
A localized infection occurred in a Harris hawk present- 2. Kasantikul T, Sommanustweechai A, Polsrila K, et al: Retrospec-
tive study on fatal melioidosis in captive zoo animals in Thailand,
ing with a necrotizing inguinal dermatitis. A pure culture
Transbound Emerg Dis 2015.
of B. pseudomallei (ST70) was grown from the lesion. The 3. Mollaret HH: l’affaire du jardin de splantes ou comment la meli-
animal was successfully treated with IM marbofloxacin oidose fit son apparition en france, Med Mal Infect 18:643–654,
10 mg/kg BID for 2 weeks followed by PO cefuroxime 1988.
100 mg/kg SID for 20 weeks. There is clearly still room for 4. White NJ: Melioidosis, Lancet 361:1715–1722, 2003.
more options and efficacy in the diagnosis and treatment 5. Pumpuang A, Chantratia N, Wikraiphat C, et al: Survival of
of avian melioidosis. Burkholderia pseudomallei in distilled water for 16 years, Trans R
Studies show that wild birds present a low but real risk Soc Trop Med Hyg 105(10–2):598–600, 2011.
of dispersing melioidosis.55 6. Dejsirilert S, Kondo E, Chiewslip D, et al: Growth and survival
of Pseudomonas pseudomallei in acidic environments, Jpn J Med
Sci Biol 44:63–74, 1991.
Other Species 7. Chan SY, Wuthiekanun V, Gao Y, et al: Effects of different salini-
ties on the survival of Burkholderia pseudomallei. In Proceedings
Other species, including reptiles and fish, may become of the 5th World Melioidosis Congress 2007; p 206.
infected or carry the disease.1,2 Pet iguanas (Iguana iguana) 8. Inglis TJ, Rigby P, Robertson TA, et al: Interaction between
were implicated in importing B. pseudomallei in California Burkholderia pseudomallei and Acanthamoeba species results in
and in the Czech Republic.56,57 coiling phagocytosis, endamebic bacterial survival, and escape,
Infect Immun 68:1681–1686, 2000.
Conclusion 9. Levy A, Chang BJ, Abbott LK, et al: Invasion of spores of the
arbuscular mycorrhizal fungus Gigaspora decipiens by Burkhold-
In conclusion, global increase in awareness has multiplied eria spp, Appl Environ Microbiol 69:6250–6256, 2003.
the number of reports and expanded the known distribu- 10. Inglis TJ, Sagripanti JL: Environmental factors that affect the
tion and impact of melioidosis in humans and animals. survival and persistence of Burkholderia pseudomallei, Appl
Environ Microbiol 72:6865–6875, 2006.
Management of zoological collections in endemic areas 10a. Martelli P, Hui SW: Ocean Park guideline of laboratory opera-
requires dedicated veterinary and husbandry practices. tions, environmental laboratory, updated May 2017.
Preventative measures against melioidosis such as species 11. Limmathurotskul D, Dance D, Wuthiekanun V, et al: Systematic
selection, enclosure design, sanitation practices, and specific review and consensus guildelines for environmental sampling
staff training are important to minimize the impact of meli- of Burkholderia pseudomallei, PLoS Negl Trop Dis 7(3):e2105,
oidosis. There are no effective vaccines against melioidosis 2013.
320 SE C T I O N 9 Infectious Diseases
12. Kaestli M, Mayo M, Harrington G, et al: Sensitive and specific 31. Lau SKP, Tang BSF, Curreem SOT, et al: Matrix-assisted laser
molecular detection Burkholderia pseudomallei, the causative desorption ionization time of flight mass spectrometry for
agent of melioidosis, in the soil of tropical northern Australia, rapid identification of Burkholderia pseudomallei: importance of
Appl Environ Microbiol 73:6891–6897, 2007. expanding databases with pathogens endemic to different locali-
13. Lau SKP, Chan SY, Currem SOT, et al: Burkholderia pseudomallei ties, J Clin Microbiol 50:3142–3143, 2012.
in soil samples from an oceanarium in Hong Kong detected using 32. Wang H, Chen YL, Teng SH, et al: Evaluation of the Bruker
a sensitive PCR assay, Emerg Microbes Infect 3:e69, 2014. Biotyper matrix-assisted laser desorption/ionization time-of-
14. Lee P, Chan SY, Hui SW, et al: Introduction of live/dead screen- flight mass spectrometry system for identification of clinical
ing system for Burkholderia pseudomallei PCR in soil sample. and environmental isolates of Burkholderia pseudomallei, Front
In Proceedings of the 7th World Melioidosis Congress 2013; Microbiol 7:2016. article 415.
p 170. 33. Anuntagool N, Naigowit P, Petkanchanapong V, et al: Monoclonal
15. Godoy D, Randle G, Simpson A, et al: Multilocus sequence antibody-based rapid identification of Burkholderia pseudomallei
typing and evolutionary relationships among the causative in blood culture fluid from patients with community-acquired
agents of melioidosis and glanders, Burkholderia pseudomallei septicaemia, J Med Microbiol 49(12):1075–1078, 2000.
and Burkholderia mallei, J Clin Microbiol 41:2068–2079, 2003. 34. Chantratita N, Tandhavanant S, Wongsuvan G, et al: Short
16. McCombie RL, Finkelstein RA, Woods D: Mutilocus sequence report: Rapid detection of Burkholderia pseudomallei in blood
typing of historical Burkholderia pseudomallei isolates collected cultures using a monoclonal antibody based immunofluorescent
in Southeast Asia from 1964 to 1967 provides insight into the assay, Am J Trop Med Hyg 89(5):971–972, 2013.
epidemiology of melioidosis, J Clin Microbiol 44:2951–2962, 35. Houghton R, Reed D, Hubbard M, et al: Development of a
2006. prototype lateral flow immunoassay (LFI) for the rapid diagnosis
17. Chan SY, Tsui HH, Hui SW, et al: Characterization of Burkhold- of melioidosis, PLoS Negl Trop Dis 8(3):e2727, 2014.
eria pseudomallei isolates from animals and environment using 36. Robertson G, Sorenson A, Govan B, et al: Rapid diagnostics for
multilocus sequence typing at an oceanarium in Hong Kong. In melioidosis: a comparative study of a novel lateral flow antigen
Proceedings of the 6th World Melioidosis Congress 2010; p 66. detection assay, J Med Microbiol 64:845–848, 2015.
18. Thomas AD, Forbes-Faulkner JC, Norton JH, et al: Clinical and 37. Lertmemongkolchai G, Khaenam P, Norton R: The molecular
pathological observations on goats experimentally infected with detection of Burkholderia pseudomallei. In Ketheesan N, editor:
Pseudomonas pseudomallei, Aust Vet J 65(2):43–46, 1988. Melioidosis: a century of observation and research, Amsterdam,
19. McCornmick JB, Sexton DJ, McMurray JG, et al: Human-to- 2012, Elsevier, pp 168–173.
human transmission of Pseudomonas pseudomallei, Ann Intern 38. Alexander AD, Huxsoll DL, Warner ARJ, et al: Serological
Med 83:512–513, 1975. diagnosis of human melioidosis with indirect hemagglutination
20. McBride J, Currie BJ: Transmission of Burkholderia pseudomal- and complement fixation tests, Appl Microbiol 20:825–833,
lei via breast milk in northern Australia, Pediatr Infect Dis J 1970.
23:1169–1171, 2004. 39. Chow KE: Development and evaluation of indirect enzyme-
21. Abbink FC, Orendi JM, de Beaufort AJ: Mother-to-child linked immunosorbent assays for diagnosis of melioidosis in
transmission of Burkholderia pseudomallei, N Engl J Med marine mammals at Ocean Park. BS (Veterinary) Thesis, Uni-
344:1171–1172, 2001. versity of Sydney, Australia, 2004.
22. Noyal MJC, Harish BN, Bhat V, et al: Neonatal melioidosis: 40. Puthucheary SD, Vadivelu J: Human melioidosis, 2002, Singapore
a case report from India, Indian J Med Microbiol 27:260–263, University Press, p 19.
2009. 41. Yang H, Kooi CD, Sokol PA: Ability of Pseudomonas pseudomal-
23. Lumbiganon P, Pengsaa K, Puapermpoonsiri S, et al: Neonatal lei malleobactin to acquire transferrin-bound, lactoferrin-bound,
melioidosis: a report of 5 cases, Pediatr Infect Dis J 7:634–636, and cell-derived iron, Infect Immun 61:656–662, 1993.
1988. 42. Garcia-Parraga D, Gilabert JA, Valls M, et al: Pharmacokinetics
24. Ko WC, Cheung BMH, Tang HJ, et al: Melioidosis outbreak of cefovecin (Convenia) after intramuscular administration to
after typhoon, Southern Taiwan, Emerg Infect Dis 13(6):896–898, dolphins (Tursiops truncates) and sea lion (Otaria flavescens). In
2007. Proceedings of 41st IAAAM Conference, 2010; p 42.
25. Kinoshita RE: Epidemiology of melioidosis in an oceanarium: a 43. Tse L: An evaluation of the indicative value of acute phase
clinical, environmental and molecular study. M.Phil. Thesis for proteins in captive seals and sea lions, naturally infected with
the University of Hong Kong, 2003. Burkholderia pseudomallei. M.Sc. Thesis for the Hong Kong
26. Liu X, Pang L, Sim SH, et al: Association of melioidosis inci- Polytechnic University, 2010.
dence with rainfall and humidity, Singapore, 2003–2102, Emerg 44. Martelli P, Fernando N, Lee FK, et al: Placement of a central
Infect Dis 21(1):159–161, 2015. indwelling catheter in a trained seal lion (Zalophus califor-
27. Kinoshita RE: Melioidosis in marine mammals. In Zoo and wild nianus) for the purpose of intensive therapy in the treatment
animal medicine current therapy (vol 6). 2008, pp 299–307. of melioidosis. In Proceedings of 42th IAAAM Conference,
28. Hoffmaster AR, AuCoin D, Baccam P, et al: Melioidosis diag- 2011.
nostic workshop 2013, Emerg Infect Dis 21(2):1–9, 2015. 45. Currie BJ, Fisher DA, Howard DM, et al: The epidemiology
29. Lau SKP, Sridhar S, Ho CC, et al: Laboratory diagnosis of of melioidosis in Australia and Papua New Guinea, Acta Trop
melioidosis: past, present and future, Exp Biol Med 240:742–751, 74:121–127, 2000.
2015. 46. Tammemagi L, Johnston AY: Melioidosis in an orangutan in
30. Karger A, Stock R, Ziller M, et al: Rapid identification of north Queensland, Aust Vet J 39:241–242, 1963.
Burkholderia mallei and Burkholderia pseudomallei by intact cell 47. Theobald V, Mayo M, Benedict S, et al: Melioidosis in exotic
matrix-assisted laser desorption/ionization mass spectrometric animals from a tropical wildlife park. In Proceedings of the 8th
typing, BMC Microbiol 12:229, 2012. World Melioidosis Congress 2016, poster 96.
CHAPTER 46 Update on Melioidosis in Zoo and Wild Animals 321
48. Yap EH, Thong TW, Tan AL, et al: Comparison of Pseudomonas 53. Sommanustweechai A, Kasanntikul T, Somsa W, et al: Environ-
pseudomallei from humans, animals, soil and water by restriction mental management procedures following fatal melioidosis in a
endonuclease analysis, Singapore Med J 36:60–62, 1995. captive chimpanzee (Pan troglodytes), J Zoo Wildl Med 44(2):475,
49. Inglis TJJ: The treatment of melioidosis, Pharmaceuticals 2013.
3:1296–1303, 2010. 54. MacKnight K, Chow D, See B, et al: Melioidosis in a macaroni
50. Lipsitz R, Garges S, Aurigemma R, et al: Workshop on treat- penguin Eudyptes chrysolophus, Dis Aquat Org 9:105–107, 1990.
ment of and postexposure prophylaxis for Burkholderia pseudo- 55. Hampton V, Kaestli M, Mayo M, et al: Melioidosis in birds and
mallei and B. mallei infection, 2010, Emerg Infect Dis 18(12): Burkholderia pseudomallei dispersal, Australia, Emerg Infect Dis
2012. 17(7):1310–1312, 2011.
51. Parkes HM, Shilton CM, Jerrett IV, et al: Primary ocular melioi- 56. Zehnder AM, Hawkins MG, Koski MA, et al: Burkholderia
dosis due to a single genotype of Burkholderia pseudomallei in two pseudomallei isolates in 2 pet iguanas, California USA, Emerg
cats from Arnhem Land in the Northern Territory of Australia, Infect Dis 20(2):304–306, 2014.
J Feline Med Surg 11:856–863, 2009. 57. Elschner MC, Hnizdo J, Stamm I, et al: Isolation of the highly
52. Kongmakee P, Chaisongkram C, Sannanu S, et al: The situa- pathogenic and zoonotic agent Burkholderia pseudomallei from
tion on melioidosis in Thailand zoos. In Proceedings of the 9th a pet green iguana in Prague, Czech Republic, BMC Vet Res
International Meeting of ASCM. 2016; p 41. 10:283, 2014.
SECTION 10
Aquatic
47 Techniques for Addressing Parasites in Saltwater
Aquariums, 323
322
47
Techniques for Addressing Parasites in
Saltwater Aquariums
CLAIRE ERLACHER-REID
P
arasitic infections of fish are common and are often definitive diagnosis, humane euthanasia and necropsy
secondary opportunistic infections; therefore it is on a subset of fish, including wet mount examination of
recommended to establish a thorough quarantine pro- internal organs and histopathology, may be considered a
tocol (Box 47.1) in efforts to reduce the risks of introducing reasonable diagnostic tool. Other diagnostics that may be
parasitic infections to an established aquarium system.1–4 pursued to evaluate for the presence of parasites in some
Occasionally, breakthrough parasitic infestations still occur, saltwater fish species include fecal examination, freshwater
despite an intensive quarantine protocol, due to parasite dip, endoscopy, and coelomic saline flush. Identification of
life cycle, anatomic location of parasite on affected animal, the parasite to species or genus is not always required to
or treatment failure.5 Implementing a thorough routine determine an initial therapeutic course of action.6
preventative medicine protocol (Box 47.2) may facilitate
diagnosis and guide treatment and/or management deci- Treatment
sions for parasitic infections of established fish collections
(see also Chapter 49). The sensitivity of skin, fin, and gill wet mount diagnostic
examinations may be low in subclinical or minimally affected
Diagnostics fish because the acquired sample represents only a small
portion of the actual tissue; therefore a negative test result
Diagnostics should first consist of evaluation of the does not ensure that the population of fish examined are
environment, water quality, and visual observation of the truly parasite free.7 For this reason, prophylactic treatments
fish. Clinical signs of parasitic infections may be non- may be used as part of a quarantine protocol to prevent the
specific and may include the following: abnormal/erratic introduction of parasites into an established system.3,4
swimming, flashing (i.e., rubbing body across surfaces), Numerous factors must be taken into consideration when
clamped fins (i.e., holding fins close to body), increased deciding on a treatment plan besides the target parasite.
respiratory rate and/or piping (i.e., gulping at surface), color Environmental factors (e.g., temperature, salinity, dissolved
changes to skin and/or gills, excessive mucus on skin and/ oxygen, alkalinity) may influence treatment and monitor-
or gills, raised nodules, ulcerative and/or erosive lesions ing. It is important to know, for example, that for each
on the skin, loss of scales and/or fins, exophthalmia and/ 5 mg/L of formalin added to a system, it depletes 1 mg/L of
or cloudy eyes, coelomic distension, emaciation, or gross oxygen.1 Thus, increasing aeration and closely monitoring
visual observation of large external parasites. The majority dissolved oxygen are essential when treating with formalin,
of fish diseases can be diagnosed by evaluation of water especially at high temperatures. The decision to treat indi-
quality and wet mount examination of tissues with light vidual fish or an entire aquarium system/population often
microscopy.6 Therefore skin scrapes, fin clips, and gill biopsy guides the veterinarian’s medication selection and desired
wet mount examination with light microscopy should be route of administration. When oral medications are chosen,
considered the minimum diagnostic database for most the animal’s current appetite and the food preference of
species of fish when presented for a physical examination. the species typically guides the formulation selected (e.g.,
To avoid potential parasitic detachment that could decrease bioencapsulation in brine shrimp, medicated flakes or gel,
the sensitivity of your diagnostic test, when possible, it gruel). The life cycle of the target parasite influences treat-
is best to acquire these samples prior to sedation, while ment frequency and duration. Oviparous parasitic species
ensuring human and animal safety are still prioritized.6 usually require multiple repeated treatments and a longer
When there is high morbidity and mortality in a collection duration of treatment when compared with viviparous
of fish and ante mortem diagnostics have not revealed a parasitic species. Furthermore, it is important to know
323
324 SE C T I O N 10 Aquatic
• BOX 47.1 Quarantine Considerations for Fish • BOX 47.2 Routine Preventative Medicine
Maintained in Public Display Aquaria Considerations for Fish Maintained in
Public Display Aquaria
All new incoming fish species should be placed in quarantine for
a minimum of 30 days (45–60 days usually recommended for Elasmobranchs
cold water species).
Routine Health Evaluations
Entrance and Exit Examinations At minimum all elasmobranchs should be visually examined
by a staff veterinarian annually. When feasible, handling an
Entrance and exit examinations on a subset of the population elasmobranch for a brief physical examination once a year is
(~2%–5%) should occur within 1 week of quarantine arrival advised and should be discussed among staff veterinarians and
and 1 week prior to quarantine departure. The following aquarium leadership. The decision to handle should be based
criteria should be met when possible during both of those on species, human and animal safety, enclosure and ability to
examinations: catch, available equipment, and available space. If handled for
• Visual inspection a physical examination, the following sampling criteria should be
• Body weight met when possible:
• Skin scrape (+/− fin clip) wet mount examination • Appropriate restraint for select species to ensure both animal
• Gill clip wet mount examination when feasible and human safety (e.g., tonic clonic immobilization, oxygen
• Blood sampling for hematology and plasma biochemistry, narcosis, behavioral or manual restraint in a net or sling, or
especially in moray eel and elasmobranch species (not chemical)
recommended in fish <8 cm total length) • Body weight and morphometric measurements
• Thorough necropsy of all deceased fish when possible • Blood sampling for hematology, plasma chemistries, and
including postmortem wet mount samples of skin scrapes, fin point-of-care analyzer (e.g., blood gasses and lactate)
clips, gill biopsies, and all internal organs • Skin scrape and gill biopsy wet mount examination when
possible
Treatments • Ultrasound examination, especially females
Treatment protocols should be designated by veterinary staff. • Coelomic wash for species considered susceptible to Eimeria
This may include prophylactic treatment of quarantined fish southwelli (e.g., cownose rays)
to prevent parasitic introduction to established systems (e.g., • Radiographs for select species (e.g., sand tiger sharks with
praziquantel, formalin, fenbendazole, copper) and/or specific suspected spinal deformity)
treatment based on diagnostic examinations.
Teleost Fish
Medical Records Routine Health Evaluations
Written or computerized records should be maintained for each At minimum all teleost fish should be visually examined by a
system documenting the following criteria daily to monitor trends staff veterinarian annually. If a fish is handled for any cause,
during the quarantine period: the following minimal physical examination criteria should be
• Water quality parameters performed when feasible:
• Number of mortalities • Appropriate restraint for select species to ensure both animal
• Treatments administered (e.g., dose, duration/frequency, and and human safety (e.g., manual restraint in a net/sling or
route) chemical)
• Estimated percentage of food intake for the population • Weight and appropriate morphometric measurements
• Skin scrape (+/− fin clip) wet mount examination
Biosecurity • Gill clip wet mount examination when feasible
• Fish should ideally be quarantined in an isolated facility • Blood sampling for hematology and plasma biochemistry
located away from collection animals when possible (not recommended in fish <8 cm total length)
• Teleost fish, elasmobranchs, and invertebrate species should
be quarantined in separate systems by taxa when possible
• There should be designated staff members assigned to
work only in the quarantine facility. Alternatively, if that is the consequences the selected treatment may have on the
not possible, collection fish should be handled first prior to filtration, microbe community, and plants in the exhibit
employees entering the quarantine facility for the day and species-specific sensitivities to the medication prior to
• Foot baths should be maintained at all quarantine entryways
and exits, and if possible, between systems
treating. If this knowledge is not known, it is advised to
• Designated nets and equipment should be assigned to each consult with experienced colleagues, treat a smaller number
system. Alternatively, if that is not possible, gloves, hand of fish, or use surrogate species and monitor them closely
washing stations, and net disinfection stations should be prior to treating an entire system or population to avoid
readily available and easy to access unexpected adverse results.1
This information was compiled and summarized from previous published Treatment for parasitic infections may consist of environ-
literature on fish quarantine protocols; therefore the reader is encouraged to mental manipulation (e.g., temperature or salinity changes),
refer to these references for complete details.3,4
the addition of biological control (e.g., cleaner fish), manual
removal of parasites, and chemical medications. It is usually
recommended to treat the entire aquarium environment,
when possible and when applicable, for the best chance of
successful therapy.1 However, once parasites are introduced
to an established system, treatment may become problematic,
especially when involving large mixed-species aquariums
CHAPTER 47 Techniques for Addressing Parasites in Saltwater Aquariums 325
with complex life support systems. When that does occur, drug concentrations throughout treatment to determine
efforts are usually focused on parasitic management by main- appropriate dosage frequencies and to ensure therapeutic
taining animal health and appropriate temperature, water levels are maintained. Removing fish from an experienced
quality, filtration, substrate, and stocking density for the system into a naïve system or enclosure for each treatment
housed aquarium species to prevent parasitic flare-ups and may help to minimize rapid degradation.
subsequent morbidity and mortality. Indefinite hyposalinity
(Table 47.1) has been a valuable tool for the successful man- Capsalid Management in a Large Mixed-
agement of Neobenedinia sp. in large mixed-species exhibits Species Saltwater Aquarium
containing elasmobranchs and teleost fish. There has also
been evidence to suggest that short-term use of hyposalinity, Capsaloidea or capsalids, including the genera Neobenedenia
along with temperature increases, may aid in resolution and Benedenia, are large oviparous monopisthocotylean
of Cryptocaryon irritans infections in large tropical marine flatworms (approximately 3–10 mm) commonly found in
exhibits as well (K. Heym, personal communication, March tropical saltwater aquariums.6 They may infest the skin,
16, 2017). In cases of parasitic flare-ups in which a specific gills, and eyes of teleost fish and elasmobranchs, causing
species is markedly compromised and the entire system flashing, erratic swimming, scale loss, erosions of the skin,
cannot be treated, it may become necessary to remove those and ulcerations of the skin and eyes, and subsequently may
fish from the environment and treat individually. Manual lead to death. Diagnosis may be made with gross observa-
removal of parasites, short-term dips and baths, medications tion of the parasites, wet mount examination of the skin
administered via gastric lavage, and parenteral treatment are and gills, or histopathology. The parasites may also easily be
often reserved for treating individual fish or anorectic fish. recovered from the bottom of a bucket or tank following
Because parasitic infections are often secondary and oppor- a freshwater dip, and this method is often used to reduce
tunistic, it is important to complete a thorough physical parasitic loads in heavily parasitized fish prior to further
examination on heavily parasitized fish to determine if other treatments. Capsalids produce many triangular-shaped
medications (e.g., antimicrobials) and supportive care are eggs (more than 80 in a day in some species) that may
needed in addition to antiparasitic agents. take 4–21+ days to hatch depending on environmental
temperature.6 These eggs also contain long sticky threads
Challenges and Novel Treatment that are used for attachment to fish, substrate, nets, and
other objects. For these reasons, they are extremely difficult
Considerations to eliminate from a system once established, and efforts are
Degradation of Praziquantel and Formalin in usually focused on management to reduce outbreaks.
a Recirculating System Chemical treatments are often difficult in large aquarium
systems due to cost, volume, differences in species sen-
Research has discovered significant degradation of both sitivities, difficulty maintaining therapeutic levels, and
praziquantel and formalin treatments in saltwater aquariums bacterial degradation of the medication. Chronic exposure
when using standard published doses and frequencies.8,9 to reduced salinity is thought to reduce egg viability and
Failure to maintain therapeutic concentrations in a system may be a useful tool in the management of capsalids in large
could lead to decreased efficacy, recurrence of pathogen, aquarium systems. Indefinite hyposalinity (see Table 47.1)
and possibly the development of resistance.10 The microbial has proven successful at preventing recurrent outbreaks of
population within the aquarium system is thought to be external monogeneans (Neobenedenia sp.) in a 500,000-
the primary contributing factor for the degradation of gallon mixed-species exhibit housing sea turtles, sand tiger
these treatments. Degradation rates appear to increase with sharks (Carcharias taurus), nurse sharks (Ginglymostoma
subsequent dosing of these medications, possibly related to cirratum), southern stingrays (Dasyatis americana), spiny
increased bacterial activity or bacterial growth that may use lobsters (Panulirus argus), and teleost fish (K. Heym, per-
these medications as an energy source after the first exposure. sonal communication, March 16, 2017). Hyposalinity has
Investigations have demonstrated that an initial dose of been maintained in this exhibit since 2011, with no obvious
praziquantel at 2 mg/L and formalin at 25 mg/L degraded adverse effects observed in the collection species. Only when
to less than detectable limits in naïve recirculating saltwater attempts were made to increase the salinity did outbreaks
systems in approximately 9 days and 14 hours, respectively. and resultant morbidity and mortality reoccur. Biological
The rate of removal for each of those medications contin- control with cleaner fish may also be a viable solution for
ued to increase with subsequent treatments at those same reducing parasitic load in large aquarium exhibits.6
doses.8,9 Similar degradation has been observed in clinical
settings, with praziquantel reaching nondetectable levels in Leech Infestation in a Large Mixed-Species
as little as 8 hours in a freshwater aquarium (M. Hyatt, Saltwater Aquarium
personal communication, February 27, 2017) and in as
little as 4–6 hours in a saltwater aquarium (S. Boylan, Inadvertent introduction of Branchellion torpedinis leeches
personal communication, February 27, 2017). These into closed saltwater aquariums has reported. These marine
investigations emphasize the significance of monitoring Text continued on p. 331
TABLE
47.1 Treatment Protocols for Commonly Diagnosed Parasites in Saltwater Public Aquaria
Motile Ciliates irritans skin or gills; be attained slowly over 3 days6 requiring addition of
histopathology Formalin: hyposalinity and/or transfer
• 25 mg/L (0.025 mL/L) EOD up to 4 weeks27 to clean aquarium and/or
• 125–250 mg/L (0.125–0.25 mL/L) for 60 min q 3 days6,28 temperature manipulations
Chloroquine diphosphate: in conjunction with
• 10 mg/L prolonged bath for 2–3 weeks27 chemical txs
• 10 mg/L q 5 days for four doses6 Copper, in the form of copper
Hyposalinity: sulfate, is typically the tx of
• 14–18 g/L for 21–30 days. Salinity should be reduced slowly by 5–10 g/L a day. choice
• 15–18 g/L for 42–56 days along with temperature increases in large aquariums
housing elasmobranchs, teleosts, and sea turtles may aid in resolution of
infection (K. Heym, personal communication, March 16, 2017)
• 10 g/L for 3 h q 3 days for 4 txs6,27
Transfer fish to clean aquarium between chemical txs or transfer to a clean
aquarium q 3 days6
Temperature manipulations: Peak reproduction occurs between 20°C and 30°C
requiring frequent and repeated medication dosing. Reproduction stops at 19°C6
Uronemia spp. Wet mount of Metronidazole: When restricted to external
Scuticociliatosis skin, gills, • Bath at 50 mg/L daily for 10 days reportedly was successful in treating an surface of fish, organisms
or internal outbreak of scuticociliatosis (Philasterides dicentrarchi) in Australian Pot-bellied are easily treated. When
organs; Seahorses24 organisms invade deep
histopathology • 50 mg/kg PO SID for 7 days followed 30 days later with ponazuril at 10 mg/kg muscles, internal organs,
PO SID for 3 days, and repeated in 2 weeks may have been helpful in reducing and blood vessels, there is
parasitic load in a zebra shark with subcutaneous scuticociliatosis (S. DiRocco, a poor prognosis6
personal communication, March 2, 2017) Philasterides dicentrarchi
Jenoclean (Atacama extract 97% [Zeolites] + citric acid 3%) at 50 mg/L for 30 min and potentially Uronema
to treat scuticociliatosis (Philasterides dicentrarchi) in Olive Flounder (Paralichthys spp. reportedly pathogenic
olivaceus)29 in sea dragons and
Hydrogen Peroxide at 50 mg/L for 30 min to treat scuticociliatosis (Philasterides seahorses23–25
dicentrarchi) in Olive Flounder29
Formalin:
• 200 mg/L (0.2 mL/L) formalin bath for 2 h SID for 6 days used in Japanese
flounder6
• 125–250 mg/L (0.125–0.25 mL/L) for 1 h may be helpful to clear early superficial
skin infestation6,28
• 25 mg/L (0.025 mL/L) 24 h bath repeated EOD for 2–3 txs may be helpful to
clear superficial skin infestation. May be used as an adjunct to a freshwater
bath6,28
Other: Improve husbandry
Trichodina spp. Wet mount of Improve water quality and husbandry Often an indicator of
skin or gills; Formalin: poor sanitation or
histopathology • 25 mg/L (0.025 mL/L) 24 h bath repeated EOD for 2–3 txs if needed6,28 overcrowding30
• 125–250 mg/L (0.125–0.25 mL/L) 1-hour bath repeated SID for 2–3 days if
needed6,28
Cu2+ at 0.15–0.2 mg/L of free Cu2+ until effect6,28
Others: Freshwater dip/bath
Brooklynella spp. Wet mount of Formalin: Often not susceptible to
skin or gills; • 125–250 mg/L (0.125–0.25 mL/L) 1 h bath repeated SID for 2–3 days if needed6 Cu2+6
histopathology • 25 mg/L (0.025 mL/L) 24 h bath repeated EOD for 2–3 txs if needed6
Protozoa: Amyloodinium Wet mount of Cu2+ at 0.15–0.2 mg/L prolonged immersion for 2–3 weeks31 Chloroquine recently tx of
External ocellatum skin or gills; Chloroquine diphosphate at 10 mg/L prolonged immersion once or redose in 7–8 choice but Cu2+ still most
Dinoflagellate histopathology days for 2–4 txs if needed6 widely used
Hydrogen Peroxide at 75 mg/L (0.21 mL of 35% H2O2/L) for 30 min, retreat after 6 Can infest both
days and immediately move fish to an uncontaminated system6 elasmobranchs and
Others: Temperature manipulation, hyposalinity, freshwater baths/dips teleosts6
Clownfish seem to be
particularly susceptible to
this parasite30
Protozoa: Ichthyobodo spp. Wet mount of Formalin: Parasite has a direct life
External skin or gills; • 25 mg/L (0.025 mL/L) 24 h bath repeated EOD if needed6,28 cycle; therefore a single tx
Flagellate histopathology • 125–250 mg/L (0.125–0.25 mL/L) 1 h bath repeated SID if needed6,28 is usually effective
Metronidazole at 40 g/kg of feed at 2% BW per day for 10 days6
Secnidazole at 20 g/kg of feed per day at 2% BW for at least 2 days6
Triclabendazole at 40 g/kg of feed per day at 2% BW for at least 5 days6
Other: Improve husbandry/sanitation
Protozoa: Eimeria southwelli Wet mount of Cu2+ wire particles (50 mg, Copasure) at a dose of 36.7 mg/kg administered PO Associated with morbidity
Coccidia coelomic once appears to be the current tx of choice16 and mortality in cownose
saline flush; Toltrazuril at 10 mg/kg/day PO for 5 days may help to control but does not rays14
histopathology eliminate infection30
Metazoan: Monopisthocoty- Wet mount of Praziquantel: Oviparous species require
Monogeneans leans skin or gills; • 2–10 mg/L immersion for 3–6 h bath, repeat in 7 days6,28 multiple repeated txs (≥3)
Gyrodactylidae gross visual • 2–3 mg/L weekly for 4–5 txs used in conjunction with Cu2+ tx for oviparous at appropriate intervals
(viviparous) observation monogeneans (R. George, personal communication, January 27, 2017) and a longer duration of
Dactylogyridae of large • 8 mg/L prolonged immersion for 20 days, redose every fourth day tx when compared with
(oviparous) monogeneans; • 20 mg/L immersion for 1.5 h6 viviparous species
Capsalidae histopathology • 100 mg/L bath for 4 min or 20 g/kg of feed q 48 h at 1% BW were both Reproductive rate is
(oviparous) effective in reducing polyopisthocotylea in rockfish32 controlled by temperature
Monocotylidae • 5 mg/L immersion for 1,800 min (30 h), 10 mg/L immersion for 120 min, or (typically faster at warmer
(oviparous) 20 mg/L for 45–90 min reportedly reduces monogenean loads on eagle rays but temperatures)6
Polyopisthocotylea does not eliminate12,13 (R. George, personal communication, February 20, 2017)
(oviparous)
Continued
CHAPTER 47 Techniques for Addressing Parasites in Saltwater Aquariums
327
TABLE
47.1 Treatment Protocols for Commonly Diagnosed Parasites in Saltwater Public Aquaria—cont’d
• 5 mg/L praziquantel prolonged immersion for 1,800 min (30 h) in a naïve Praziquantel is generally the
treatment tank, moving animals to a second naïve treatment tank, then tx of choice; however,
treating with 3 mg/L praziquantel q 5 days for 4 to 5 months in conjunction it is strongly advised
with 10 mg/L chloroquine has been utilized successfully by at least one facility to measure drug levels
to treat monogeneans in eagle rays such that all life stages ceased to be in the water to confirm
detected after 2 months of treatment. Chloroquine concentration should begin therapeutic levels are
at 3 mg/L and slowly increase by 2–3 mg/L EOD until a final concentration being maintained. This
of 10 mg/L is achieved. No obvious adverse effects were noted in eagle rays knowledge may then
or Atlantic stingrays at this facility; however, cownose rays demonstrated be used to decide the
changes in swimming behavior during the treatment and were removed. It is frequency of repeated or
also recommended to remove ozone from the treatment tanks because it may redosed txs
react with the chloroquine and cause inappetance or other adverse effects in Capsalids have an affinity for
elasmobranchs (R. George, personal communication, February 20, 2017). ocular tissue
• 100 mg/kg BW via oral gavage reportedly reduces monogenean loads in eagle Dacylogyrids and
rays but does not eliminate. More research is currently underway5 Polyopisthocotylea have
• 100 mg/kg BW split into four doses SID for 3 days6 an affinity for gill tissue. Gill
Cu2+ 0.15 mg/L of free Cu2+ for 3–4 months to treat oviparous monogeneans. monogeneans are often
Treatment concentration should be attained slowly over 3 days. Often used as more resistant to treatment
an adjunct to praziquantel (3 mg/L weekly for 4–5 txs). Has been used without than skin parasites6
adverse effects in cownose rays but has caused inappetence in southern and Dactylogyroidea are most
Atlantic stingrays (R. George, personal communication, January 27, 2017) commonly found in
Hyposalinity: freshwater fish
• <20 g/L may reduce egg viability in oviparous species6
• Indefinite hyposalinity (20–24 ppt) has proven successful at preventing recurrent
outbreaks of external monogenean (Neobenedenia sp.) parasites in a large
mixed species exhibit (500,000 gallons) housing teleosts, elasmobranchs, and
sea turtles (K. Heym, personal communication, March 16, 2017)
Formalin:
• 150–250 mg/L (0.15–0.25 mL/L) bath for 60 min6,28
• 15–25 mg/L (0.015–0.025 mL/L) prolonged immersion6,28
Trichlorfon (Dylox):
• 2–5 mg/L bath for 60 min6
• 0.25–1 mg/L prolonged immersion for 2 tx at 3-day intervals for dactylogyrid
species.6 Has been used as an adjunct treatment prior to praziquantel use.
Mebendazole:
• 100 mg/L bath for 10 min6
• 1 mg/L prolonged immersion for 24 h6
Biological control: French angel fish, neon gobies, cleaning gobies, and blue-lined
cleaner wrasse pick monogeneans off other fish6,33
Other: Freshwater dip/bath to reduce parasite load, chloramine T
Metazoan: Wet mounts of Prevention: exclude birds and mammals from contact with fish, disinfect and Most common in freshwater
Digeneans gill, skeletal quarantine, intermediate host usually mollusk wild fish. Uncommon
muscle, Praziquantel: in aquaria due to the
intestinal • 1 mg/L immersion for 90 h6 complex life cycle involving
contents, or • 2–10 mg/L for 24 h prolonged immersion6 a variety of host animals
other organs; • 10 mg/L bath for 1 h6 Self-limiting and does not
histopathology • 25 mg/kg BW IM once6 progress in aquariums
• 50 mg/kg BW PO once6 without exposure to hosts
• 330 mg/kg BW PO once6
Other: Cu2+, Slaked lime, and Bayluscide molluscicides
Metazoan: Wet mount of Praziquantel: Tx targets adults not larvae
Cestodes intestinal • 5 mg/kg BW PO q 7 days up to 3 txs28 Do not feed live foods that
content or • 50 mg/kg BW PO once6 might transmit larval
other internal • 2 mg/L bath for 1–3 h, repeat in 1 week if needed6 cestodes
tissues/ Other: Exclude intermediate host contact with fish and water supply
organs;
histopathology
Metazoan: Wet mount Fenbendazole: Tx targets adults, not larvae.
Nematodes of feces, • 2 mg/L once a week for 3 weeks6,28 Encysted nematodes are
intestinal • 2.5 mg/g of food daily for 3 days, repeat in 2–3 weeks28 difficult to treat
contents, or • 25 mg/kg BW PO for 3 days6 Fish may be definitive hosts
other internal • 50 mg/kg BW PO once a week for 2 txs6 for adult nematodes or
tissues/ Levamisole: act as an intermediate
organs; • 4 g/kg of food once a week for 3 weeks28 host or transport for larval
histopathology • 2.5–10 mg/kg BW PO SID for 7 days28 nematodes
• 10 mg/kg BW PO once a week for 3 weeks28 Most nematodes are
• 10 mg/kg BW PO once, then repeated in 14 days resolved Huffmanela sp. oviparous, but there are
lesions in sandbar sharks34 some that are viviparous
• 10 mg/kg BW IM once, repeated at 14 and 28 days. Cleared Huffmanela sp. and some with direct life
egg tracks from a sandbar shark (Carcharhinus plumbeus)34 cycles
• 1–2 mg/L immersion for 24 h, repeat in 2–3 weeks28 Ivermectin has been used but
• 10 mg/L bath for 3 days28,30 is not recommended due
Other: Piperazine, trichlorphon, and mebendazole. Avoid feeding organisms to fish to low therapeutic index30
that may harbor the larvae (e.g., live copepods are a common source)
Metazoan: Gross visual Trichlorfon (Dylox) at 0.25–0.4 mg/L for a 5–6 h bath was used successfully to treat Premedication with atropine
Leeches observation Branchellion torpedinis leeches in elasmobranchs. Repeated tx in 30 days may at 0.04–0.08 mg/kg IM is
(Annelids) or wet mount be needed (A. McDermott, personal communication, January 26, 2017). It is advised (A. McDermott,
of skin, gills, important to remember when calculating the dose that commercial preparations personal communication,
and oral cavity of organophosphates vary in percentage of active ingredients6 January 26, 2017)
samples; Spotted eagle rays
histopathology anecdotally appear more
sensitive to this treatment;
therefore, pre-medication
with ≥0.06 mg/kg of
atropine is highly advised
Organophosphates are
typically the tx of choice
CHAPTER 47 Techniques for Addressing Parasites in Saltwater Aquariums
Continued
329
TABLE
47.1 Treatment Protocols for Commonly Diagnosed Parasites in Saltwater Public Aquaria—cont’d
Crustaceans Branchiurans observation • 0.01 mg/L immersion for 48 h q 6 days for 3 txs28 the most effective tx for
Isopods or wet mount • 0.03 mg/L immersion for Lernaea spp.6 crustacean parasites30
of gills, skin, • 75 mg/kg BW PO for 14 days for sea lice6 Enamectin has been
or oral cavity Enamectin: associated with rare
samples; • 50 µg/kg BW PO for 7 days for sea lice6,28 neurotoxicity and death
histopathology • 2 mg per 100 g of dry gel food, add water, and mix to create a medicated gel. when a fish ingests a
Feed approximately 2% BW for an estimated fish intake of 400 µg/kg/day for 10 greater amount of food
days for treatment of copepods (L. Adams, personal communication, March 14, than anticipated. Reduce
2017) the dose for ravenous
Trichlorfon: fish (L. Adams, personal
• 15–300 mg/L for 15–60 min at 3–18°C for sea lice6 communication, March 14,
• 2–5 mg/L for 60 min for isopods6 2017)
• 0.25–1 mg/L immersion28 It is important to remember
Dichlorvos: when calculating trichlorfon
• 0.5–2 mg/L for 30–60 min for sea lice6 doses that commercial
• 15 mg/L for 1 min for sea lice6 preparations often vary
Hydrogen Peroxide at 1250 mg/L (1.25 mL of 50% H2O2/L) for up to 30 min bath6 in percentage of active
Formalin at 150–250 mg/L (0.15–0.25 mL/L) immersion for 60 min6,28 ingredients6
Carbaryl (1-napthyl N-methylcarbamate), available as a suspension (Sevin 225 mg/ Carbaryl treatment may
mL) has been used as an alternative to trichlorfon for successful treatment of cause anorexia and at
isopods, branchurians, and copepods in temperate marine fish (<18°C) and a dose of 0.5 mg/L has
freshwater eels: resulted in muscle tetany
• 0.25–0.35 mg/L for 4–5 days or spasms in fish but has
• 0.25–0.35 mg/L for 2 txs at 48 h intervals with water changes in between (L. resolved 24 h following
Adams, personal communication, March 14, 2017) removal of the medication
Milbemycin oxime (Interceptor) alone or with spinosad (Triflexis) has been used for (L. Adams, personal
treatment of copepods: communication, March 14,
• 0.5–1 mg/kg PO once 2017)
• 4 mg/100 g of gel food (0.004%) fed one day (L. Adams, personal
communication, January 27, 2017)
Other: Manual removal, biological control, and freshwater dip/bath
The listed doses have been acquired from both published and anecdotal references and may not be appropriate for all species in all water conditions. Many of the listed treatments are not yet supported with
pharmacokinetic data; therefore the reader is encouraged to use caution. If adverse reactions are observed, fish should be removed from the treatment immediately. BW, Body weight; EOD, every other day; h, hour(s);
IM, intramuscular; min, minutes; PO, orally; q, every; SID, once daily; tx(s), treatment(s).
CHAPTER 47 Techniques for Addressing Parasites in Saltwater Aquariums 331
leeches exclusively parasitize elasmobranchs, resulting in to remove ozone before treatment because it is believed
ulcerations at attachment sites, lethargy, anorexia, anemia, to react with chloroquine and cause inappetance or other
and potential death in as little as 5 days (A. McDermott, adverse effects in elasmobranchs (R. George, personal com-
personal communication, January 26, 2017).6,11 Leeches munication, February 20, 2017). Praziquantel administered
may also serve as vectors for infectious diseases. Affected via gastric gavage to anesthetized spotted eagle rays has
species have included sawfish (Pristis pristis), guitarfish also resulted in dramatic decreases in parasite loads but
(Rhina ancylostoma), zebra sharks (Stegostoma fasciatum), did not eliminate (see Table 47.1).5 Research investigating
spotted eagle rays (Aetobatus narinari), manta rays (Manta pharmacokinetic data for this oral treatment regimen and
birostris), southern stingrays, and experimentally yellow various other chemical immersions along with additional
stingrays (Urobatis jamaicensis) (A. McDermott, personal management options are currently ongoing.
communication, January 26, 2017).11 Leeches are most
commonly recovered from the claspers, pectoral fins, eyes, Eimeria southwelli in Cownose Rays
oral cavity, and cephalic lobes and appear to remain perma- (Rhinoptera bonasus)
nently attached to the host if not removed. Manual removal
when leeches are easily accessible is a treatment option, Eimeria southwelli are apicomplexa coccidia parasites that
but the process is time consuming. Trichlorfon (Dylox) may be associated with morbidity and mortality in cownose
for a 5- to 6-hour bath has also been used with success; rays at high numbers.14 In small numbers and in the absence
however, it is strongly recommended to premedicate with of clinical signs, this parasite might be considered normal
atropine approximately 45–60 minutes prior to trichlorfon flora that may clear over time without treatment.15 Clinical
treatment (see Table 47.1). It is also important to remember signs have included discoloration of the skin, emaciation,
when calculating the dose that commercial preparations and death. Diagnosis includes microscopic identification of
of organophosphates vary in percentage of active ingredi- organisms on a wet mount obtained from a coelomic saline
ents.6 Leech cocoons will hatch in approximately 30 days; flush. A coelomic flush may be obtained in one of two ways:
therefore repeated treatment may become necessary at that (1) gently passing a lubricated sterile red rubber catheter
time.11 Topical or systemic antibiotic administration and (3–5 Fr) attached to a syringe through one of the coelomic
blood transfusions have also been helpful in the support- pores found on either side of the cloaca or (2) inserting
ive care and recovery of the affected host when deemed a winged infusion set attached to a syringe (21-gauge,
necessary by the veterinarian. Future studies to establish 19-mm needle) into the right ventral paramedian body
pharmacokinetic effects and safety for trichlorfon in various wall cranial to the pelvic girdle. Sterile 0.9% saline may
species of elasmobranchs are warranted. be infused into the coelomic cavity at 1% of the animal’s
body weight and aspirated for microscopic examination.14
Monogeneans in Spotted Eagle Rays The current most successful treatment when clinical signs
(Aetobatus narinari) are apparent appears to be copper wire particles (Copasure)
administered orally once (see Table 47.1).16 No obvious
Decacotyle floridana and Clemacotyle australis are monocot- negative effects have been documented to date, and the
ylid monogenean parasites that have been associated with animals do not appear to excrete copper to detectable levels
morbidity and mortality in spotted eagle rays.5,12,13 These in their enclosure following treatment (E. Clarke, personal
parasites have been recovered from the skin and gills of communication, January 26, 2017). Further studies to
spotted eagle rays, and clinical signs have included abnormal establish pharmacokinetic effects and safety for copper
swimming postures, bottom resting, and rubbing on the wire particles in elasmobranchs are needed. Toltrazuril,
walls of the enclosure.12 Praziquantel immersion (see Table ponazuril, clindamycin, and sulfadimethoxine treatments
47.1) reportedly reduces parasite loads and is often used as a administered at various doses, frequencies, and routes have
management tool; however, this treatment does not success- been attempted with inconsistent or inconclusive results. A
fully eradicate the parasite, requires repeated therapy and preliminary study evaluating the use of ponazuril in cownose
frequent handling, and is further complicated by bacterial rays did not achieve blood concentrations considered to
degradation of the medication in the water.12,13 A multistep be therapeutic in other species, but the parasite load did
treatment protocol involving a prolonged immersion of decrease and there were no negative side effects observed
praziquantel in a naïve treatment tank for 30 hours followed (S. Cassle, personal communication, January 24, 2017).
later by immersion with chloroquine in combination with Cloacal prolapse may be a complication of the parasitic
praziquantel for 4–5 months in a second naïve treatment infection and/or treatment attempts associated with the
tank (see Table 47.1) was utilized successfully by at least infection.17
one facility to treat monogeneans in eagle rays such that
all life stages ceased to be detected. No obvious adverse Copper Immersion in Cownose Rays
effects were noted in eagle rays or Atlantic stingrays with (Rhinoptera bonasus)
this treatment protocol at this facility; however, cownose
rays demonstrated changes in swimming behavior during The use of copper immersion treatment has typically been
this treatment and were removed. It is also recommended avoided in elasmobranchs due to intolerance and mortalities
332 SE C T I O N 10 Aquatic
associated with respiratory, osmoregulatory, and ionoregula- brain, and/or liver. It is thought that this disease may have
tory distress.3,15,18–21 However, it has been used safely as a been underreported or unrecognized in elasmobranchs until
prolonged immersion in cownose rays for 3–4 months in now or is an emerging disease.22 Systemic invasion has been
conjunction with praziquantel prolonged immersion weekly associated with a poor prognosis. To date, there are no known
for 4–5 treatments (see Table 47.1) to treat capsalid parasite effective treatment protocols published in elasmobranchs.
infestations (R. George, personal communication, January However, treatment with oral metronidazole followed a
27, 2017). Closely monitoring copper and praziquantel con- month later with oral ponazuril may have been helpful in
centrations in the water is extremely important throughout clearing or reducing parasitic load in a zebra shark with
the course of treatment. Although no adverse effects have subcutaneous scuticociliatosis (see Table 47.1). Follow-up
been observed in cownose rays, it has been associated with on the case to monitor for recurrence is currently ongoing
inappetence in southern and Atlantic stingrays (Dasyatis (S. DiRocco, personal communication, March 2, 2017).
sabina) (R. George, personal communication, January 27,
2017). Pharmacokinetic studies to evaluate the effects and Summary
safety of copper immersion at various concentrations in
elasmobranchs are warranted. Prevention and early diagnosis of parasitic infections are
recommended by establishing a thorough quarantine and
Scuticociliatosis in Sygnathid and preventative medicine protocol whenever possible. There
Elasmobranch Species are a multitude of published and unpublished protocols
that have been used for the treatment and management
Philasterides dicentrarchi are ciliated protozoa in the subclass of parasitic infections in fish with variable success; there-
Scuticociliatia that have been identified as the cause of disease fore the reader is strongly encouraged to use caution and
outbreaks in a range of marine teleost fish in aquarium and perform a thorough literature search and discuss treatment
aquaculture settings.22 Severe outbreaks of this parasite have options with experienced colleagues to select the treat-
recently been reported in aquarium-maintained Australian ment protocol that is best suited for each specific situa-
pot-bellied seahorses (Hippocampus abdominalis), weedy tion (see Table 47.1). There are numerous considerations
sea dragons (Phyllopteryx taeniolatus), and leafy sea dragons that should be addressed prior to and during treatment,
(Phycodurus eques) and may be associated with stressful including potential species-specific sensitivities, life cycle
environmental conditions (e.g., temperature fluctuations, of the parasite, and the relationship between and among
poor water quality, transport) resulting in compromise of water quality parameters, parasites, fish, and treatments.
the immune system. Clinically these animals have presented The reader is encouraged to monitor drug concentrations
with nodular or ulcerative epidermal lesions, hyperemia throughout treatment to determine appropriate dosage
or depigmentation, anorexia, lethargy, irregular respira- frequencies and ensure therapeutic levels are maintained.
tions, abnormal swimming, and/or death. Histopathologic Further pharmacokinetic research evaluating the effects and
examination revealed ciliate invasion into the gills, dermis, safety of antiparasitic medications in various species of fish
subdermal connective tissues, vasculature, skeletal muscle, are greatly needed in the available peer-reviewed literature.
ovary, kidney, intestines, thyroid, and/or brain.23–25 Treat- The World Association for the Advancement of Veterinary
ment with a prolonged immersion of metronidazole (see Parasitology (WAAVP) has created guidelines for testing the
Table 47.1) for 10 days appeared to be successful in treating efficacy of ectoparasiticides in finfish, and this may serve as
systemic disease in Australian pot-bellied seahorses; however, a useful resource for designing meaningful studies.35
knowledge of additional treatment options for sygnathids
is lacking in the literature, and further pharmacokinetic
research is warranted.24
References
Philasterides dicentrarchi has also been the cause of a 1. Harms CA: Treatments for parasitic diseases of aquarium and
rapidly lethal systemic infection in aquarium-maintained ornamental fish, Sem Avian Exotic Pet Med 5:54–63, 1996.
zebra sharks, Port Jackson sharks (Heterodontus portusjack- 2. Harms CA: Fish. In Fowler ME, Miller RE, editors: Zoo and wild
soni), and Japanese horn sharks (Heterodontus japonicus) animal medicine, ed 5, St. Louis, MO, 2003, Elsevier, pp 2–20.
characterized by necrotizing hepatitis, meningoencephalitis, 3. Hadfield CA, Clayton LA: Fish quarantine: current practices in
and/or thrombosing branchitis. Clinical signs prior to death public zoos and aquaria, J Zoo Wildl Med 42:641–650, 2011.
were brief and included lethargy, anorexia, and/or behavioral 4. Hadfield CA: Quarantine of fish and aquatic invertebrates in
abnormalities.22 Ciliate infections resembling scuticociliates public display aquaria. In Miller RE, Fowler ME, editors: Zoo
and wild animal medicine: current therapy (vol 7). St. Louis, 2012,
have also been identified in swell sharks (Cephaloscyllium
Elsevier, pp 202–209.
ventriosum), dusky smooth-hounds (Mustelus canis), south- 5. MyIniczenko ND, Nolan EC, Thomas A, et al: Management
ern stingrays, and California bat rays (Myliobatis californica) of monogenes in eagle rays (Aetobatus narinari) with high dose
(C. Erlacher-Reid, unpublished, 2016).22,26 These infections oral praziquantel, in Proceedings of the 48th annual IAAAM
have varied in severity, ranging from external infections of Conference, 2015.
the gills and skin, along with bacterial, flagellate, or viral 6. Noga EJ: Fish disease: diagnosis and treatment, ed 2, Ames, IA,
infections, to deep invasive lesions involving the kidney, 2010, Wiley-Blackwell.
CHAPTER 47 Techniques for Addressing Parasites in Saltwater Aquariums 333
7. Larrat S, Marvin J, Lair S: Low sensitivity of antemortem gill 23. Bonar CJ, Garner MM, Weber ES, et al: Pathological findings
biopsies for the detection of subclinical Pseudodactylogyrus bini in weedy (Phyllopteryx taeniolatus) and leafy (Phycodurus eques)
infestations in American eels (Anguilla rostrata), J Zoo Wildl Med seadragons, Vet Pathol 50:368–376, 2013.
43:190–192, 2012. 24. Cicco ED, Paradis E, Stephen C, et al: Scuticociliatid ciliate
8. Knight SJ, Boles L, Stamper MA: Response of recirculating salt- outbreak in Australian pot-bellied seahorse, Hippocampus
water aquariums to long-term formalin treatment, J Zoo Aquar abdominalis (Lesson, 1827): clinical signs, histopathologic
Res 4:77–84, 2016. findings, and treatment with metronidazole, J Zoo Wildl Med
9. Thomas A, Dawson MR, Ellis H, et al: Praziquantel degradation 44:435–440, 2013.
in marine aquarium water, PeerJ 4:e1857, 2016, doi:10.7717/ 25. Rossteuscher S, Wenker C, Jermann T, et al: Severe scutico-
peerj.1857. ciliate (Philasterides dicentrarchi) infection in a population of sea
10. Kuemmerer K: Antibiotics in the aquatic environment: a review. dragons (Phycodurus eques and Phyllopteryx taeniolatus), Vet Pathol
Part II, Chemosphere 75:435–441, 2009. 45:546–550, 2008.
11. Marancik DP, Dove AD, Camus AC: Experimental infection 26. Garner MM: A retrospective study of disease in elasmobranchs,
of yellow stingrays (Urobatis jamaicensis) with the marine leech Vet Pathol 50:377–389, 2013.
(Branchellion torpedinis), Dis Aquat Organ 101:51–60, 2012. 27. Yanong RPE: Cryptocaryon irritans infections (marine white
12. Janse M, Borgsteede FHM: Praziquantel treatment of captive spot disease) in fish. Program in Fisheries and Aquatic Sciences,
white-spotted eagle rays (Aetobatus narinari) infested with mono- SFRC, Florida Cooperative Extension Service, Institute of Food
gean trematodes, Bull Eur Ass Fish Pathol 23:152–156, 2003. and Agricultural Sciences, University of Florida, Gainesville,
13. Nolan EC, MyIniczenko ND, Steinmetz J, et al: Oral praziqu- FL, 2009. Retrieved 1 March 2017 from http://edis.ifas.ufl.edu/
antel pharmacokinetics in spotted eagle rays (Aetobatus narinari), fa164.
in Proceedings of the 47th annual IAAAM Conference, 2016. 28. Lewbart GA: Fish. In Carpenter JW, editor: Exotic animal
14. Stamper MA, Lewbart GA: Eimeria southwelli infection associ- formulary, ed 4, St. Louis, MO, 2013, Elsevier, pp 18–52.
ated with high mortality of cownose rays, J Aquat Anim Health 29. Jin C-N, Harikrishnan R, Moon Y-G, et al: Effectiveness of che-
10:264–270, 1998. motherapeutants against scuticociliate Philasterides dicentrarchi, a
15. MyIniczenko ND. Medical management of rays. In Miller RE, parasite of olive flounder, Vet Parasitol 168:19–24, 2010.
Fowler ME, editors: Zoo and wild animal medicine: current 30. Petty BD, Francis-Floyd R: Parasitic diseases of fish. In Aiello
therapy (vol 7). St. Louis, 2012, Elsevier, pp 170–176. SE, Moses MA, editors: The Merck veterinary manual, ed 11, NJ,
16. Clarke EO, Grzenda K, Murphy D, et al: Successful treatment 2016, Merck & Co., Inc.
of Eimeria southwelli in a cownose ray (Rhinoptera bonasus) using 31. Reed P, Francis-Floyd R: Amyloodinium infections of marine fish.
oral copper wire particles, in Proceedings of the 46th annual College of Veterinary Medicine, Florida Cooperative Extension
IAAAM Conference, 2016. Service, Institute of Food and Agricultural Sciences, University
17. McDermott A, Field C, Mejia-Fava J, et al: Medical and surgical of Florida, Gainesville, FL, 1994. Retrieved 1 March 2017
management of cloacal prolapses in cownose rays (Rhinoptera from http://agrilife.org/fisheries/files/2013/09/Amyloodinium-
bonasus) undergoing treatment for Eimeria southwelli infection, Infections-of-Marine-Fish.pdf.
in Proceedings of the 44th annual IAAAM Conference, 2013. 32. Kim KH, Cho JB: Treatment of Microcotyle sebastis (Monogenea:
18. Boeck GD, Hattink J, Franklin NM, et al: Copper toxicity in Polyopisthocotylea) infestation with praziquantel in an experimen-
the spiny dogfish (Squalus acanthias): urea loss contributes to the tal cage simulating commercial rockfish Sebastes schlegeli culture
osmoregulatory disturbance, Aquat Toxicol 84:133–141, 2007. conditions, Dis Aquat Organ 40:229–231, 2000.
19. Grosell M, Wood CM, Walsh PJ: Copper homeostasis and 33. Cowell LE, Watanabe WO, Head WD: Use of tropical cleaner
toxicity in the elasmobranch Raha erinacea and the teleost fish to control the ectoparasite Neobenedenia melleni (Monogenea:
Myoxocephalus octodecemspinosus during exposure to elevated Capsalidae) on seawater cultured Florida red tilapia, Aquaculture
water-borne copper, Comp Biochem Physiol C Toxicol Pharmacol 113:189–200, 1993.
135:179–190, 2003. 34. MacLean RA, Fatzinger MH, Woolard KD, et al: Clearance
20. Grosell M: Intestinal anion exchange in marine fish osmoregula- of a dermal Huffmanela sp. in a sandbar shark (Carcharhinus
tion, J Exp Biol 209:2813–2827, 2006. plumbeus) using levamisole, Dis Aquat Organ 73:83–88, 2006.
21. Grosell M, Blanchard J, Brix KV, et al: Physiology is pivotal for 35. Sommerville C, Endris R, Bell TA, et al: World association for
interactions between salinity and acute copper toxicity to fish and the advancement of veterinary parasitology (WAAVP) guideline
invertebrates, Aquat Toxicol 84:162–172, 2007. for testing the efficacy of ectoparasiticides for fish, Vet Parasitol
22. Stidworthy MF, Garner MM, Bradway DS, et al: Systemic 219:84–99, 2016.
scuticociliatosis (Philasterides dicentrarchi) in sharks, Vet Pathol
51:628–632, 2014.
48
Touch-Pools: The Other Side of
the Hand
CATHERINE HADFIELD AND KATHRYN A. TUXBURY
334
CHAPTER 48 Touch-Pools: The Other Side of the Hand 335
Planning
In the authors’ experience, the design, construction, and
commissioning of touch-pools often takes longer than
display habitats. All groups that are involved in building and
managing the habitat should be involved in the planning
process, including education and media representatives.
Additional time for animal training prior to moving to B
the habitat, as well as once in the habitat, is important in • Figure 48.1 Touch-pools demonstrating wide areas for refugia and
meeting goals. This can be accomplished by slowly increas- bays for staff access (A) and larger system with mangrove décor for
ing guest numbers in a controlled manner. A soft opening additional refugia (B). (A Courtesy National Aquarium; B Courtesy New
provides a gradual acclimation and time for environmental England Aquarium.)
or management changes. Staff and volunteer training for
touch-pools is also time-consuming but is essential for that extend into the habitat or a central dry space. The
animal care, guest management, and education goals. habitat shape may create narrow sections; it is important
The space required by a touch-pool is greater than that that those sections are wide enough to allow animals to
required by a display-only habitat because of the habitat pass through freely. In large touch-pools, interpreters may
shape, additional life-support equipment needs, and guest enter the habitat in waders or wetsuits, although this may
routing based on peak attendance and anticipated stay-times. be a more stressful scenario for the animals and should
The shape of the habitat must provide refugia, some- always be preceded by a cue.7 Accessibility and visibility
times known as retreat spaces; this is a fundamental welfare to the public are important factors. Concrete tanks with
requirement.6 These are spaces that are out of reach of guests acrylic windows are preferred because they provide better
and potentially out of sight. They provide the opportunity sound-dampening.8 Large windows may increase the need
for each animal to control its interactions with humans. for refugia that animals can choose to use.
The ideal refugia would be part of the habitat that extends The design of the interior of the habitat is also important
into a space that is separated from the guest space. More for animal health. Substrate and décor that is suitable for the
commonly, refugia are still in sight of guests but out of target species must be included to meet the behavioral needs
direct touch, for example, wide or deep sections (>18 inches of the animals. This may include sand, rocks, small coral
or 50 cm) that guests cannot reach, or caves or tunnels structures, mangrove roots, and/or sea grasses. Substrate
(Fig. 48.1). For touch-pools where guests have access on all may also reduce noise exposure for resident animals.8
sides, the habitat should be large enough to provide ample The life-support system of a touch-pool often needs
refugia in the center. To maximize the impact of refugia, to be larger or more complex than a display habitat of
staff should not catch animals from those areas. a similar volume to manage possible contaminants (e.g.,
For good animal care and guest interactions, the habitat skin oils, lotions, sunscreen, insect repellant, perfume, and
shape should allow interpreters to have access to the human hair). Water turnover must be high (more than once
animals and a view of the guests. This may involve bays per hour) with excellent surface skimming. The biological
336 SE C T I O N 10 Aquatic
filtration needs to accommodate the maximum animal Other scenarios should be considered with any exhibit:
bioload as well as the possible contaminants, based on peak • a catastrophic life-support failure (e.g., pump failure
attendance. In saltwater systems, good foam fractionation decreasing dissolved oxygen);
is essential. Both ultraviolet light and ozone disinfection • significant morbidity or mortality;
are strongly recommended. Where elasmobranchs are in a • a visible animal abnormality, which may represent a
system using ozone, the levels of nitrate, iodate, and iodide genuine health concern or a cosmetic issue that guests
should be monitored to avoid goiters.9 may notice;
It is essential for welfare reasons that animals are provided • welfare concerns.
with a regular, suitable period of full darkness. This may It is extremely helpful to be able to close off a touch-pool
conflict with late night events, custodial needs, and safety area and stop guest access if needed for animal care.
lighting requirements, and needs should be discussed prior
to construction. To help maintain appropriate temperature, Monitoring of Touch-Pools
the heating, ventilation, and air-conditioning (HVAC)
system must be able to cope with regular changes in guest Monitoring of touch-pool habitats and animals should be
numbers as well as seasonal temperature and humidity similar to display-only habitats, but closer tracking may be
fluctuations. required to ensure that health and welfare standards are met.
As with any exhibit, the authors have found that animal Water quality parameters must be assessed regularly. This
health and welfare are improved with the provision of suit- should include temperature, dissolved oxygen, pH, salin-
able off-exhibit holding space for all the species. However, ity, nitrogenous wastes, calcium, alkalinity, and hardness.
this may be even more important for a touch-pool where Touch-pools may show fluctuations in dissolved oxygen,
animals may benefit from rotations off-exhibit. Holding temperature, and ammonia based on guest numbers, par-
space should be in a quiet area close to the touch-pool ticularly in shallow areas and after peak guest attendance.
to minimize transport time. Having the holding space on In-line monitoring of dissolved oxygen and temperature are
the same water system as the exhibit allows animals to helpful to monitor trends, particularly around animal intro-
be transferred without an acclimation, but a water quality ductions, peak guest attendance, and any changes to the
issue on exhibit will then affect the holding space. The best life-support system. Water quality monitoring may include
option may be a combination of holding space that is on the bacterial counts. These are typically run based on indica-
same water system and holding space that is on a separate tor bacteria tests (e.g., total and fecal coliform bacteria).13
water system nearby. These may not represent changes in pathogenic bacteria,
Hand-washing and drying stations at the touch-pool but after a baseline is established for a specific system, they
entrance could reduce contamination of the system, but can be used to determine the level of disinfection required
compliance is likely to be low and it is probably more effec- to maintain the baseline.
tive to rely on the life-support system. Best practices dictate Feeding and behavior records should be reviewed
that hand-washing and drying stations are provided after regularly, particularly because touch-pool animals are often
touch-pools to reduce the risk of zoonotic disease transmis- managed by a wider range of staff than display habitats of
sion. The zoonotic risk is low, particularly compared with a similar size. Feed records, weights, and body condition
terrestrial animal interactive experiences such as petting scores should be used to evaluate feeding needs on a routine
zoos.10 However, with young guests and an increasing basis. Behaviors that should be tracked include bite wounds,
proportion of immunosuppressed people, services should mating, or stereotypies. If animals are exhibiting undesir-
always be provided to reduce the risks. Hand-cleaning sta- able behaviors, environmental and behavioral modification
tions should be easily visible, always available, on an exit plans should be instituted quickly. For example, aggression
route, and accessible at different heights. Signage should may be reduced by providing increased foraging opportuni-
be clear and readily visible. It may help to have digital ties. However, individuals or species that are not thriving in
screens to attract guests to the stations. Running water, a touch-pool and not responding to the changes provided
foaming hand wash, and hot-air or jet-air drying are usually should be transferred to different habitats where they can
recommended, although hand-sanitizers are associated with thrive.
greater compliance.11 Compliance is still poor (typically Routine examinations are often indicated to assess health
30%–60% at petting zoo hand-washing stations) but is and welfare criteria. This may consist of visual or hands-on
improved by verbal reminders by staff at the exit who are examinations with appropriate diagnostic tests. Routine
actively dispensing hand sanitizer.11,12 trimming of the venomous barb in stingrays is common in
Contingency planning is essential with any new exhibit. touch-pools. These events can provide unique educational
Some scenarios are specific to touch-pools: opportunities.
• contamination by human bodily fluids (e.g., blood or Animal health and welfare criteria should be assessed
vomit); continuously and reviewed on a routine basis. All touch-
• human injury (e.g., from a stingray barb, an urchin pool animals, vertebrate and invertebrate, should do as
spine, a bite, or a fall); well as conspecifics in display-only habitats. It is likely
• a zoonotic question or complaint. that it will take 2–3 years to find the optimal balance of
CHAPTER 48 Touch-Pools: The Other Side of the Hand 337
animal, life-support, and guest dynamics, at which point 3. Bassett L, Buchanan-Smith HM: Effects of predictability on the
the population and management should be well established. welfare of captive animals, Appl Anim Behav Sci 102:223–245,
2007.
4. Claxton AM: The potential of the human-animal relationship
Staff and Guest Considerations as an environmental enrichment for the welfare of zoo-housed
animals, Appl Anim Behav Sci 133:1–10, 2011.
Touch-pools are typically maintained by a combination 5. Laule GE: Positive reinforcement training and environmental
of animal husbandry and guest experience/education staff, enrichment: enhancing animal well-being, J Am Vet Med Assoc
some of whom are often volunteers. All staff should have 223:969–973, 2003.
extensive training on the natural history of the various 6. Anderson US, Benne M, Bloomsmith MA, et al: Retreat
species, conservation and education goals, and common space and human guest density moderate undesirable behavior
questions that arise. Unusual facts and “juicy questions” in petting zoo animals, J Appl Anim Welfare Sci 5:125–137,
encourage guest interaction and may be more effective than 2002.
scripted talks.14 Staff must also have a solid understanding of 7. Chisholm LA, Whittington ID, Fischer ABP: A review of
animal touch rules. Instructions are often that animals may Dendromonocotyle (Monogenea: Monocotylidae) from the skin
of stingrays and their control in public aquaria, Folia Parasitol
be gently touched with two fingers but not moved or picked
51:123–130, 2004.
up. Written signs may help, but their wording is important 8. Anderson PA: Acoustic characterization of seahorse tank environ-
and signs are not always read.15,16 Although guests have good ments in public aquaria: a citizen science project, Aquacult Eng
intentions, they may adversely affect the animals through 54:72–77, 2013.
lack of direction, lack of experience, and enthusiasm (e.g., 9. Morris AL, Stremme DW, Sheppard BJ, et al: The onset of goiter
moving, manipulating, or grabbing animals). Staff should in several species of sharks following the addition of ozone to a
be trained in effective guest control, with a focus on positive touch-pool, J Zoo Wildl Med 43:621–624, 2012.
reinforcement of preferred behavior. 10. Steinmuller N, Demma L, Bender JB, et al: Outbreaks of enteric
Evaluation of the guest metrics is important to determine disease associated with animal contact: not just a foodborne
if the touch-pool goals are being met. This may include guest problem anymore, Clin Infect Dis 43:1596–1602, 2006.
numbers, time spent at touch-pools, number of interpreter 11. Anderson MEC, Weese JS: Video observation of hand hygiene
practices at a petting zoo and the impact of hand hygiene inter-
interactions, and guest evaluations.1,14,17
ventions, Epidemiol Infect 140:182–190, 2012.
12. Erdozain G, KuKanich K, Chapman B, et al: Observation of
Conclusion public health risk behaviours, risk communication and hand
hygiene at Kansas and Missouri petting zoos – 2010–2011,
The immersive experiences provided by touch-pools can Zoonoses Public Health 60:304–310, 2012.
foster excitement about animals and conservation. With 13. Culpepper EE, Clayton LA, Hadfield CA, et al: Coliform bacte-
good planning and management, touch-pools can meet ria monitoring in fish systems: current practices in public aquaria,
animal health and welfare, education, and conservation J Aquat Anim Health 28:85–90, 2016.
goals. Examples of goals may include meeting or exceeding 14. Kopczak C, Fisiel JF, Rowe S: Families talking about ecology at
AZA animal care standards, morbidity and mortality rates touch tanks, Env Ed Res 21:129–144, 2015.
equal to nontouch conspecifics, and a sustainable manage- 15. Clayton S, Fraser J, Saunders CD: Zoo experiences: conversations,
connections, and concern for animals, Zoo Biol 28:377–397,
ment plan. Animals can thrive and be ambassadors for
2009.
change. 16. Kratochvil H, Schwammer H: Reducing acoustic disturbances
The authors would like to encourage staff at zoos and by aquarium guests, Zoo Biol 16:349–353, 1997.
aquariums to objectively assess touch-pool habitats and to 17. Wagner K, Chessler M, York P, et al: Development and imple-
contribute data to the literature. Studies exist on guest mentation of an evaluation strategy for measuring conservation
effects on mammals and birds, as well as effects of fish and outcomes, Zoo Biol 28:473–487, 2009.
aquatic invertebrate touch-pools on families.1,18–20 There are 18. Farrand A, Hosey G, Buchanan-Smith HM: The guest effect in
abundant opportunities to study guest effects on health and petting zoo-housed animals: aversive or enriching?, Appl Anim
behavior of fish and aquatic invertebrates, as well as water Beh Sci 151:117–127, 2014.
quality, morbidity and mortality comparisons, and how 19. Rowe S, Kisiel J: Family engagement at aquarium touch tanks
touch-pools affect us all. – exploring interactions and the potential for learning. In
Davidsson E, Jakobsson A, editors: Understanding interactions at
science centers and museums: approaching sociocultural perspectives,
References Rotterdam, 2012, Sense Publishers, pp 63–77.
20. Sherwen SL, Magrath MJL, Butler KL, et al: Little penguins,
1. Kisiel J, Rowe S, Vartabedian M, et al: Evidence for family
Eudyptula minor, show increased avoidance, aggression and vigi-
engagement in scientific reasoning at interactive animal exhibits,
lance in response to zoo guests, Appl Anim Beh Sci 168:71–76,
Sci Ed 96:1047–1070, 2012.
2015.
2. Sahrmann JM, Niedbalski A, Bradshaw L, et al: Changes in
human health parameters associated with a touch tank experience
at a zoological institution, Zoo Biol 35:4–13, 2016.
49
Sharks and Medicine
LEIGH CLAYTON AND KATHRYN E. SEELEY
S
harks are common and popular display animals in zoos and use some combination of ram ventilation and buccal
and aquaria. There are more than 440 known species pumping.2
of shark, with variations in lifestyle and environment. Sharks have large livers that comprise up to 25% of an
Most species are marine, with few exceptions that are not individual’s body weight. They have a simple stomach and a
typically maintained in aquaria. short small intestinal tract that empties into the spiral colon
Among the most common sharks maintained in human (also known as the spiral valve). The large surface area of
care are nurse sharks (Ginglymostoma cirratum), sand tiger the spiral colon compensates for the overall shortness of the
sharks (Carcharias taurus), sandbar sharks (Carcharhinus digestive tract and allows for more efficient nutrient absorp-
plumbeus), blacktip reef sharks (Carcharhinus melanop- tion. Sharks have a rectal gland that is located proximal to
terus), bonnethead sharks (Sphyrna tiburo), leopard sharks the cloacal opening and is thought to play a role in sodium
(Triakis semifasciata), zebra sharks (Stegostoma fasciatum), balance.2
and whitespotted bamboo sharks (Chiloscyllium plagiosum). Sharks lack bone marrow and lymph nodes and use alter-
Larger species, such as whale sharks (Rhincodon typus) and native hematopoietic organs such as the thymus, spleen,
hammerhead sharks (Sphyrnidae spp.), are less frequently epigonal organ, and Leydig cells. The ability of the epigonal
maintained but are found at some institutions.1 organ and Leydig organ to produce lymphomyeloid tissues
Sharks are in the same class as rays, both of which are is unique to elasmobranchs.2
characterized by their cartilaginous skeletons. However,
there are differences in the approach to medical manage- Husbandry and Management
ment between these elasmobranchs. This chapter will focus
on the management of shark species commonly housed in As with all aquatic animals, water quality is of paramount
aquatic and zoological institutions. importance to maintaining good health. Parameters such
as temperature, pH, salinity, nitrates, nitrite, ammonia,
Biology, Anatomy, and Physiology alkalinity, and heavy metal levels may vary between shark
taxa and should be thoroughly researched prior to bringing
The basic body plan and anatomy are similar among all a new species to an institution.
shark species. Their skeleton is composed of cartilage instead Exhibits need to be designed with both animal and
of bone, and they have small placoid scales covering their human safety in mind. Ideally an exhibit should be set up so
bodies. Sharks have a two-chambered heart consisting of an that the animal is accessible and may safely be manipulated
atrium and a ventricle. Blood is carried from the heart to or restrained. This allows for management of the individual
the capillaries in the gills, where oxygen exchange occurs. in case of illness or injury. Mixed species exhibits are pos-
The branchial arteries distribute the blood throughout the sible when size and demeanor of the species to be housed
body, and deoxygenated blood returns to the heart.2 together is taken into consideration.
Most species have five pairs of gill slits. Ventilation is
dependent on the lifestyle of the shark. Open water pelagic
sharks primarily use ram ventilation in which water is forced Restraint
over their gills while constantly swimming. Few obligate Manual Restraint
ram ventilators are kept in aquaria due to the challenge of
providing adequate space for continuous swimming in a Many small- to medium-sized sharks can be manually
captive setting. In contrast, benthic species can move water restrained with adequate facilities and experienced handlers.
over their gills through buccal pumping. This allows them The goal of any restraint is to minimize stress and ensure
to rest comfortably on the bottom and tolerate hypoxic the safety of both the animal and the handlers. The handler
environments. Many species are intermediate ventilators should wear gloves to protect from the abrasive nature of
338
CHAPTER 49 Sharks and Medicine 339
their skin. Many sharks respond well to operant condition- Physical Examination
ing and may be trained to target and voluntarily move into
a sling. Small sharks can be held safely. Many sharks go into A physical examination on a shark should begin from afar
tonic immobility when placed in dorsal recumbency. This by obtaining a thorough history and observing how the
reflex, documented in multiple species, including blacktip animal interacts with its environment and conspecifics.
reef sharks and leopard sharks, causes the individual to Appetite, attitude, swimming pattern, and evaluation of
become immobile and allows for safer handling.3 Although any external lesions may all be assessed without needing to
this restraint technique may be useful for nonpainful proce- restrain the individual.
dures, it has not been documented to have analgesic effects When performing a physical examination, the handler
and in some instances has been linked to hyperesthesia, should evaluate the shark from nose to tail. The integument
so additional analgesia should be used when performing should be assessed for any lesions, cuts, or abrasions. Overall
painful or invasive procedures.4 body condition should be evaluated, and if possible a weight
Because this method of restraint requires that the shark should be obtained. Ocular examination may be performed
must be physically turned upside down, it is applicable with the use of a light source, slit lamp, or ophthalmoscope.
only to individuals and species that are small enough to A safe evaluation of the oral cavity may be done by inserting a
do so safely. polyvinyl chloride (PVC) pipe into the oral cavity and using
Larger sharks may be restrained using a shark box by a light source to evaluate the mucosa and teeth. Respiratory
using a sling to remove them from the water and place them rate and effort should be noted by monitoring opercular
in the box. The restrainers then apply firm pressure along movement. Auscultation has little utility for monitoring
the body with one person holding the jaw shut. The box is of cardiac parameters, but a Doppler, electrocardiogram
small enough to restrict movement and allow safe sample (ECG), or ultrasound may be used to obtain heart rate
collection. It is important for all restraint that there is not a and rhythm. Because ECG clips may be difficult to place
prolonged pursuit because this may lead to elevated lactate on the skin of sharks, leads should be connected to needles
and acidemia.5 and placed in the appropriate locations. Evaluation of the
cloaca for any swelling, lesions, discharge, or discoloration
Chemical Restraint should be included in the examination. Measurements are
often taken to track growth and condition.
There are many species of shark that are too large or danger-
ous to manually restrain, and sedation or anesthesia are Diagnostics
required. Anesthetics may be administered through immer-
sion bath, injection, or orally, and numerous protocols Blood may be collected from several locations in a shark,
have been described.6,7 Equipment may vary depending depending on the species and the method of restraint. The
on species and environment, but a list of commonly used dorsal sinus and the ventral coccygeal vein are the two
materials is presented in Box 49.1. There is wide variability most commonly used sites. The ventral coccygeal vein is
in species-specific physiology. As a result, the efficacy of any preferred because blood in the dorsal sinus pools, whereas
anesthetic may depend on a number of factors, including the blood in the ventral tail vein is actively circulating.8
temperature, metabolism, drug receptors and distribution, When accessing the ventral coccygeal vein, it is important
and hepatic transformation.7 A complete review of shark to ensure that the needle is inserted at a 90-degree angle on
anesthesia is beyond the scope of this chapter, and the midline (Fig. 49.1). In many cases the needle needs to be
reader should consult additional source material for specific advanced through cartilage prior to reaching the vessel. In
protocols.7 larger species a spinal needle may be used. When obtaining
blood from the dorsal sinus the needle should be inserted
on midline caudal to the dorsal fin at a 45-degree angle
(Fig. 49.2).
Blood should always be placed immediately into antico-
• BOX 49.1 Common Equipment Needed for
agulant tubes. In sharks, dry heparin or ethylenediamine-
Shark Anesthesia
tetraacetic acid (EDTA) can both be used; however, EDTA
Appropriately sized container: tub, shark box can cause rapid hemolysis in stingray species, so if both
Stretcher species are maintained in a collection it is preferable to
Tank water for anesthesia and recovery use heparin tubes in all cases. If possible, a hemocytometer
Methods of ventilation: red rubber tubes, air pumps, catheter tip
syringes count should be performed immediately after collection
Thermometer to prevent thrombocyte and white blood cell aggregation.
pH meter If this is not possible, an aliquot can be preserved in 10%
Dissolved oxygen meter formalin for later evaluation; this maintains cell morphol-
Air stone ogy and prevents thrombocyte aggregation.9
Protective gloves
The unique physiologic characteristics of sharks must be
taken into account when interpreting blood work results.
340 SE C T I O N 10 Aquatic
Imaging
The shark skeletal system is composed of calcified car-
• Figure 49.2 Image of blood collection from the dorsal sinus in a tilage, resulting in excellent radiographic detail, making
sandbar shark (Carcharhinus plumbeus). radiographs a useful tool in evaluating skeletal structures.11
However, it is often difficult to evaluate soft tissue and
Marine elasmobranchs retain and reabsorb urea and other organs.11 When possible, both a dorsoventral and a lateral
solutes, thereby ensuring that plasma remains hyperosmotic image should be obtained. The use of contrast medium may
to their saline environment.10 Normal blood urea nitrogen be useful in evaluating the gastrointestinal tract but often
(BUN) should range from 1000 to 1300 mg/dL, and low requires multiple images.11 Because obtaining radiographs
values may indicate loss from renal disease or decreased requires removing the shark from the water, there may be
production due to hepatic disease. Marine elasmobranchs increased risk to the animal as well as potential damage to
have efficient salt excretion mechanisms in the kidney and the equipment.
specialized rectal gland that compensate for the influx of Ultrasound is very useful in assessing organ shape, loca-
sodium (Na) and chloride (Cl) from the environment. tion, and consistency and is a complement to radiology.13
However, serum concentrations of these electrolytes are It may also be used to monitor gilling and heart rates as
higher than seen in mammals. Differential counts vary well as diagnose pregnancy. It is important to note that
by species, but generally there should be 50%–75% lym- sharks have a large, lipid-filled liver, which will appear more
phocytes, 10%–30% heterophils, 0%–10% eosinophils, hyperechoic compared with mammals.
0%–1% basophils, and 0%–3% monocytes.11 Basophils Advanced imaging, such as computed tomography (CT)
are rare or absent in many shark species studied. The use of or magnetic resonance imaging (MRI), is not typically
acute phase proteins as markers of inflammation is being used in sharks due to their limited availability and com-
explored but has only been investigated in limited species.12 plicated logistics. Studies on preserved specimens have
Reference ranges have not been established in many sharks revealed that it may be a useful modality to characterize
but Tables 49.1 and 49.2 provide values for some com- internal structures.14 Due to logistic challenges, advanced
monly kept species. imaging is more often used to evaluate specific lesions of
Skin scrapes and biopsies may be taken if lesions are interest and is not generally performed as part of routine
detected on examination. It may be difficult to obtain a scanning.
CHAPTER 49 Sharks and Medicine 341
TABLE
49.1 Hematologic Reference Ranges for Select Shark Species
TABLE
49.2 Biochemical Reference Ranges for Select Shark Species
*Values too low to measure; out of range. BDL, Below detectable range.
†
Persky ME, Williams J, Burks RE, et al: Hematologic, plasma biochemistry, and select nutrient values in captive smooth dogfish (Mustelus canis). J Zoo Wildl
Med 43:842–851, 2016.
$
Haman K, Norton T, Thomas A, et al: Baseline health parameters and species comparisons among free-ranging Atlantic sharpnose (Rhizoprionodon ter-
raenovae), bonnethead (Sphyrna tiburo), and spiny dogfish (Squalus acanthias) sharks in Georgia, Florida, and Washington, USA. J Wildl Dis 48:295–306, 2012.
wild seven gill sharks (Notorynchus cepedianus), Tetraphyl- cholangiocarcinoma, testicular mesothelioma, melanoma,
lidean spp. in the spadenose shark (Scoliodon laticaudus), and lymphosarcomas have been reported.11
and Calliobothrium schneiderae in smooth-hound sharks Spinal deformities are commonly reported in captive
(Mustelus lenticulatus).24–27 Nematodes have been shown to sharks. It has been postulated that some of these deformities
cause a parasitic meningoencephalitis in nurse sharks.28 could be capture related, particularly when pound nets are
Noninfectious diseases of sharks include trauma, foreign used.29 There have also been correlations with nutritional
body ingestion, neoplasia, and deleterious environmental deficiencies, particularly potassium, zinc, and vitamin C,
impacts. Sharks may potentially ingest foreign objects that which play a critical role in cartilage development.29 In
end up in their enclosures. This is a particular consideration addition, congenital abnormalities of the vertebrae and
in aquaria where the public has access to the tank. Docu- skeletal system have been documented in several species.30
mentation of neoplastic disease is uncommon in sharks, Goiter in association with the addition of ozone to
but oral fibropapillomas, hepatic adenomas, intrahepatic a system has been reported in multiple species. When
CHAPTER 49 Sharks and Medicine 343
aquarium water is ozonated, it reduces the amount of 9. Arnold JE, Matsche MA, Rosemary K: Preserving whole blood
environmental iodine available, which is critical for thyroid in formalin extends the specimen stability period for manual cell
hormone synthesis.31 Urogenital sinus calculi have been counts for fish, Vet Clin Pathol 43:613–620, 2014.
10. Cusack L, Field CL, Hoopes L, et al: Comparison of pre- and
reported in a sand tiger shark.32
postquarantine blood chemistry and hematology values from
wild-caught cownose rays (Rhinoptera bonasus), J Zoo Wildl Med
Treatment and Therapies 47:493–500, 2016.
11. Smith M, Warmolts D, Thoney D, et al, editors: The elasmobranch
Pharmacokinetic and pharmacodynamic studies of thera- husbandry manual: Captive care of sharks, rays, and their relative,
peutics in sharks are limited. Treatment dosages are typically 2004 (https://sites.google.com/site/elasmobranchhusbandry/
extrapolated from other species. Cefovecin, a third-generation manual).
cephalosporin, maintains therapeutic serum concentrations 12. Krol L, Allender M, Cray C, et al: Plasma proteins and selected
for 4 days in white-spotted bamboo sharks (Chiloscyllium acute-phase proteins in the whitespotted bamboo shark (Chil-
plagiosum) at a dose of 8 mg/kg subcutaneously.33 A single oscyllium plagiosum), J Zoo Wildl Med 45(4):782–786, 2014.
40 mg/kg intramuscular dose of florfenicol in this species 13. Walsh MT, Pipers FS, Brendemuehl CA, et al: Ultrasonagraphy
as a diagnostic tool in shark species, Vet Radiol Ultrasound
was shown to maintain therapeutic concentrations for 120
34:213–219, 1993.
hours in serum and 72 hours in cerebrospinal fluid.34 14. Waller GNH, Williams SCR, Cookson MJ, et al: Preliminary
In cases of acute trauma and/or severe blood loss, blood analysis of elasmobranch tissue using magnetic resonance
transfusions may be performed. Cross-matching should be imaging, Magn Reson Imaging 12:535–539, 1994.
done prior to any transfusion to ensure that the donor is 15. Florio D, Gridelli S, Fioravanti ML, et al: First isolation of
compatible.35 Tenacibaculum maritimum in a captive sand tiger shark (Carch-
Nutritional support is important in anorectic animals, arias taurus), J Zoo Wildl Med 47:351–353, 2016.
and patients may be assist fed. In some animals this is done 16. Egidius E: Vibriosis: pathogenicity and pathology. a review,
by inserting whole fish into their oral cavity using tongs to Aquaculture 67:15–28, 1987.
encourage eating. If the shark does not respond to this, a 17. Grimes DJ, Colwell RR, Stemmler J, et al: Vibrio species as agents
fish gruel using their diet and vitamins may be made and of elasmobranch disease, Helgolander Meeresun 37:309–315,
1984.
tube fed. Gruel is administered by inserting a small piece
18. Marancik DP, Berliner AL, Cavin JM, et al: Disseminated fungal
of PVC into the oral cavity to pass a tube, then advancing infection in two species of captive sharks, J Zoo Wildl Med
the tube past the esophageal sphincter into the stomach. A 42:686–693, 2011.
total of 2%–5% of body weight should be given. 19. Pirarat N, Sahatrakul K, Lacharoje S, et al: Molecular and
pathological characterization of Fusarium solani species complex
References infection in the head and lateral line system of Sphyrna lewini,
Dis Aquat Organ 120:195–204, 2016.
1. American Elasmobranch Society Web site: The 2008 AES 20. Smith AG, Muhvich AG, Muhvich KH, et al: Fatal Fusarium
International Captive Elasmobranch Census. Available at: http:// solani infections in baby sharks, J Med Vet Mycol 27:83–91, 1989.
elasmo.org/census. (Accessed 19 May 2017). 21. Erlacher-Reid CD, Nollens HH, Schmitt TL, et al: Phaeohy-
2. Hamlett WC: Sharks, skates, and rays: the biology of elasmobranch phomycosis associated with ossification of the skull and cervical
fishes, Baltimore, MD, 1999, The Johns Hopkins University vertebrae in a swell shark (Cephaloscyllium ventriosum), J Zoo
Press. Wildl Med 47:1081–1085, 2016.
3. Henningsen AD: Tonic immobility in 12 elasmobranchs: use as 22. Young JM, Frasca S, Gruber SH, et al: Monogenoid infection of
an aid in captive husbandry, Zoo Biol 13:325–332, 1994. neonatal and older juvenile lemon sharks, Negaprion brevirostris
4. Mauk M, Olson R, LaHoste G, et al: Tonic immobility pro- (Carcharhinidae), in a shark nursery, J Parasitol 99:1151–1154,
duces hyperalgesia and antagonizes morphine analgesia, Science 2013.
213:353–354, 1981. 23. Bullard SA, Benz GW, Braswell JS: Dionchus postoncomiracidia
5. Hyatt MW, Anderson PA, O’Donnell PM, et al: Assessment of (monogenea: dionchidae) from the skin of blacktip sharks,
acid–base derangements among bonnethead (Sphyrna tiburo), Carcharhinus limbatus (carcharhinidae), J Parasitol 86:245–250,
bull (Carcharhinus leucas), and lemon (Negaprion brevirostris) 2000.
sharks from gillnet and longline capture and handling methods, 24. Caira JN, Durkin SM: A new genus and species of tetraphyl-
Comp Biochem Phys A 162:113–120, 2012. lidean cestode from the spadenose shark (Scoliodon laticaudus)
6. Miller SM, Mitchell MA, Heatley DJJ, et al: Clinical and in malaysian borneo, Comp Parasitol 73:42–48, 2006.
cardiorespiratory effects of propofol in the spotted bamboo 25. Ivanov VA: New species of crossobothrium (Cestoda: tetralidea)
shark (Chylloscyllium plagiosum), J Zoo Wildl Med 36:673–676, from the broadnose sevengill shark, Notorynchus cepedianus, in
2005. Argentina, J Parasitol 95:1479–1488, 2009.
7. Mylniczenko ND, Clauss TM, Stamper MA: Elasmobranchs and 26. Pickering M, Caira JN: Calliobothrium schneiderae n. sp.
holocephalans. In West G, Heard D, Caulkett N, editors: Zoo (Cestoda: tetraphyllidea) from the spotted estuary smooth-hound
animal and wildlife immobilization and anesthesia, ed 2, Ames, shark, Mustelus lenticulatus, from New Zealand, Comp Parasitol
IA, 2014, Wiley-Blackwell, pp 261–303. 75:174–181, 2008.
8. Mylniczenko N, Curtis E, Wilborn R, et al: Differences in 27. Ruhnke TR, Thompson VA: Two new species of paraoryg-
hematocrit of blood samples obtained from two venipuncture matobothrium (Tetraphyllidea: Phyllobothriidae) from the
sites in sharks, Am J Vet Res 67:1861–1864, 2006. lemon sharks Negaprion brevirostris and Negaprion acutidens
344 SE C T I O N 10 Aquatic
(Carcharhiniformes: Carcharhinidae), Comp Parasitol 73(1): 32. Walsh MT, Murru FL: Urogenital sinus calculi in a sand tiger
35–41, 2006. shark (Odontaspis taurus), J Wildl Dis 23:428–431, 1987.
28. Credille KM, Johnson LK, Reimschuessel R: Parasitic meningo- 33. Steeil JC, Schumacher J, George RH, et al: Pharmacokinetics
encephalitis in nurse sharks (Ginglymostoma cirratum), J Wildl of cefovecin (convenia®) in white bamboo sharks (Chiloscyllium
Dis 29:502–506, 1993. plagiosum) and atlantic horseshoe crabs (Limulus polyphemus), J
29. Anderson PA, Huber DR, Berzins IK: Correlations of capture, Zoo Wildl Med 45:389–392, 2014.
transport, and nutrition with spinal deformities in sandtiger 34. Zimmerman DM, Armstrong DL, Curro TG, et al: Pharma-
sharks, Carcharias taurus, in public aquaria, J Zoo Wildl Med cokinetics of florfenicol after a single intramuscular dose in
43:750–758, 2012. white-spotted bamboo sharks (Chiloscyllium plagiosum), J Zoo
30. Hoenig JM, Walsh AH: Skeletal lesions and deformities in large Wildl Med 37:165–173, 2006.
sharks, J Wildl Dis 19:27–33, 1983. 35. Hadfield CA, Haines AN, Clayton LA, et al: Cross matching
31. Morris AL, Stremme DW, Sheppard BJ, et al: The onset of goiter of blood in Carcharhiniform, Lamniform, and Orectolobiform
in several species of sharks following the addition of ozone to a sharks, J Zoo Wildl Med 41:480–486, 2010.
touch pool, J Zoo Wildl Med 43:621–624, 2012.
50
Decompression Medicine in Aquatic
Species (Fish and Sea Turtle Focus)
DANIEL GARCÍA-PÁRRAGA AND JOSÉ LUIS CRESPO-PICAZO
345
346 SE C T I O N 10 Aquatic
Recent studies reveal that capture depth and animal is observed as radiolucency (negative contrast) within or
size could have a significant impact on the likelihood of distending the vessels, or even in severe cases the cardiac
DCS.9 It was further hypothesized that variation in water chambers. Whole body DV projection generally provides
temperature, the ascent rate, the duration of entrapment, sufficient information to classify degree of GE, depending
and the type of fishery could all be important to alter on the amount and distribution of gas. Images representa-
the risk of GE.3,4 However, further research is needed tive of each category are summarized in Table 50.1. In mild
to determine which of these factors are relevant for sea cases the LL projection of the kidney region is the most
turtles. sensitive, allowing detection of relatively small amounts
From the medical point of view, forced submergence of gas. Radiographs from dead bycaught specimens
may have many other significant effects on the physiology are useful to aid with interpretation of lesions prior to
of sea turtles other than GE and DCS, such as hypoxia, necropsy.
acidosis, electrolyte unbalance, and exertional myopathy, During ultrasound examination, common findings are
among others.10–12 the presence of gas bubbles in echocardiography and/or
renal image and in severe cases, increased free fluid inside
Diagnosis the coelomic cavity. Ultrasound is the most sensitive method
to determine the presence of gas bubbles because even a
Diagnosis of DCS includes the animal’s history (type of small accumulation of gas can be seen in renal vessels as
fishing gear, duration the gear was in the water, fishing hyperechoic spots generating typical comet tail artifacts (see
depth, initial condition and progression through time), Figs. 50.1D, 50.2D, and 50.3D). However, ultrasound is
external clinical signs, and complementary diagnostic tests. less effective than radiography to estimate total volume of
Diagnostic imaging, including ultrasound, radiographs, pathologic gas and global organic distribution.
or computed tomography (CT), is the most practical and CT scan is the preferred method to quantify and discrimi-
reliable resource to accurately diagnose GE (see Chapters nate the presence of gas bubbles in different tissues, but it is
31 and 32). often not readily available and generally requires anesthesia
or at least sedation in active individuals. The whole body
Clinical History and Symptoms of the turtle should be scanned under maximum resolution
Turtles suffering from DCS show a narrow range of clinical and thinnest slice thickness to increase sensitivity and detect
signs that can be easily overlooked. Valuable information minor bubbles. CT images provide additional information
can be obtained from fishermen involved, because progres- about the presence of gas in areas where ultrasound cannot
sion of animal condition over time after surfacing seems to penetrate, such as the parenchyma of the central nervus
be especially relevant. system (CNS) and inside the vertebral canal. Bubbles in the
In our experience, bycaught turtles exhibit three main spinal cord region are frequently observed, even in mildly
behaviors right after surfacing: (1) apparently normal; (2) affected individuals (Fig. 50.4A).
hyperexcited with or without neurologic signs that may Image datasets of individuals with mild, moderate, and
develop over time (including extended neck and opened severe decompression are included (see Figs. 50.1, 50.2,
mouth, flipper retraction, limb paresis, breathing difficul- and 50.3).
ties, loss of regional nociception or superficial sensitivity, Magnetic resonance imaging (MRI) offers additional
and even full paralysis, depending on localization and prognostic information to help determine tissue damage
amount of gas bubbles); or (3) comatose. GE in various and presence of sequelae in turtles that survive. Anatomic
degrees of severity has been diagnosed following all three alterations, lack of tissue circulation/perfusion, or even
initial presentations, generally inducing a progression metabolic activity can provide information about the extent
from active to comatose in moderate to severe cases if of damage.
no specific treatment is applied. Positive buoyancy and
erratic swimming have been observed some hours after Laboratory Profile
surfacing in some individuals if returned directly to The primary acute biochemistry changes detectable in
the water. affected individuals with large presence of gas include a
moderate to severe increase in uric acid (UA) concentration
Diagnostic Imaging and a notable electrolyte imbalance (mainly hyponatremia
Combining ultrasound and radiographic examination results and hyperkalemia) at presentation. Severe elevation in
in the easiest, quickest, and most complete basic approach plasma creatine kinase (CK) and moderate elevation in
for a tentative diagnosis of GE and presumptive DCS in lactate dehydrogenase (LDH) enzyme activities are typically
sea turtles. Radiography is the simplest semiquantitative observed over the course of days following treatment. In
diagnostic tool and the most practical to categorize the addition, a left-shifted inflammatory response post treat-
severity of GE (Figs. 50.1A–C, 50.2A–C, and 50.3A–C). ment is frequently seen. However, it is difficult to discern
On the radiographs, dorsal-ventral (DV) and lateral-lateral which alterations correspond to the presence of GE versus
(LL) projections are recommended to allow a general view the other pathologic conditions that overlap with gas bubble
of the amount and distribution of gas. Intravascular gas formation during forced submergence.
CHAPTER 50 Decompression Medicine in Aquatic Species (Fish and Sea Turtle Focus) 347
* * *
A B
C D
• Figure 50.1 Imaging findings on admission in loggerhead sea turtles (Caretta caretta) with mild gas
embolism. (A) Dorsal-ventral radiographic image showing presence of gas as radiolucent angiograms
(black) on the renal veins (asterisks). (B) Computed tomography dorsal view of three-dimensional air
volume–rendering of gas spaces. Small amount of abnormal gas is clearly visualized caudal to pulmonary
silhouette. (C) Lateral-lateral radiographic image revealing intravascular gas caudal to lungs. (D) Renal
ultrasound showing gas bubbles (white arrows) evidenced as hyperechoic spots artifacts dispersed inside
kidney parenchyma and renal vessels.
cava, and other major vessels. In severe cases the spleen can
Postmortem Studies be emphysematous. Externally, the kidneys often exhibit
Ideally necropsies should be performed in the first hours multifocal extensive red areas consistent with marked con-
postmortem to avoid gas bubble formation from putrefac- gestion and acute renal infarcts. Intestinal mucosa often
tion. Special attention should be made to minimize gas reveals segmental congestive transversal bands from 1–3 cm
infiltration artifacts under traction of tissues and section wide, along different intermediate sections. The lungs of
of blood vessels (especially when removing the plastron). some animals present cranial pulmonary emphysema. Other
In mild cases, gas may be impossible to detect on necropsy gross findings include coelomic transudate in severe cases.
but is usually noticeable in moderately and severely affected Gas sampling and analysis to identify gas bubble compo-
patients. In these cases, gas often can be grossly visible sition can be performed according to previously described
in the median abdominal, mesenteric, gastric, pancreatic, methods.13 Gas composition of the bubbles should be
hepatic, and renal veins, as well as in the cardiac chambers consistent with air embolism or gases from decompression,
(especially the right atrium), the sinus venosus, the post ruling out putrefaction or gas infiltration.14
348 SE C T I O N 10 Aquatic
* *
* *
A B
C D
• Figure 50.2 Imaging findings on admission in loggerhead sea turtles (Caretta caretta) with moderate
gas embolism. (A) Dorsal-ventral radiographic image clearly showing presence of gas on renal and hepatic
veins (asterisks). (B) Computed tomography dorsal view of three-dimensional air volume–rendering of gas
spaces. Gas amount is noticeably larger than in mild cases and widely distributed over the kidney and
liver parenchyma. (C) Lateral-lateral radiographic image revealing significant intravascular gas caudal to
lungs. Lung field extension is limited due to liver gas expansion. Evident gas accumulation is present in the
caudal coelom and partially visible ventral to lungs. (D) Renal ultrasound revealing intraluminal gas in renal
parenchyma and major vessels evidenced as hyperechoic spots and comet tail artifacts. Note the loss
of detail in parenchyma due to gas bubble accumulation and comet tail artifacts in the renal portal vein.
*
*
A B
C D
• Figure 50.3 Imaging findings on admission in loggerhead sea turtles (Caretta caretta) with severe gas
embolism. (A) Dorsal-ventral radiographic image clearly showing large presence of intravascular gas in
cardiovascular system. Large vessels are filled with gas; i.e., postcava, hepatic veins, (asterisks). Lung
shape contrast is usually partially reduced or absent. (B) Computed tomography dorsal view of three-
dimensional air volume–rendering of gas spaces. Note pulmonary physiologic gas has almost disappeared
and gas is distributed following the main cardiovascular circuit. (C) Lateral-lateral radiographic image
revealing marked lung compression against the carapace due to the large presence of gas in intracoelomic
organs. (D) Renal ultrasound is impeded due to massive presence of intravascular and tissue gas.
conventional emergency treatment8,15 will help to promote diffusion through lungs, leading to successful treatment in
blood circulation and tissue perfusion and to reestablish some critical cases. Intubated animals should be closely
voluntary respiration. Stimulation of acupuncture point monitored and extubated as soon as they regain conscious-
GV26 (nasal philthrum) with a hypodermic needle also ness and start breathing voluntarily.
helps with resuscitation.16 Animals that have been through recompression therapy
Due to the simple design of some custom-made pres- should be reevaluated using radiography and ultrasound
sure chambers, animals cannot be artificially ventilated immediately after treatment, to evaluate the outcome of
when inside, so it is important to verify that sea turtles the therapy. In all cases the recompression therapy can be
are spontaneously breathing before HBOT. If respiration is repeated as necessary if some gas is still present.
interrupted, nitrogen elimination will be impeded, possibly Human-based recompression protocols were adapted to
resulting in the death of the patient inside the chamber. turtles, based on differences in anatomy and physiology.
Nonbreathing severely embolized turtles have undergone The time at elevated pressure was prolonged (up to 14–16
brief recompression cycles while intubated. This allows hours) using pure oxygen during the entire procedure.
the gas to compress, helping the nitrogen to go back into At present, an initial pressure of 2.8 ATA (1.8 ATM)
solution, reestablishing blood flow, and accelerating oxygen is maintained for the first 2 hours. The pressure is then
350 SE C T I O N 10 Aquatic
TABLE Criteria for Categorization of Gas Embolism in Live and Recently Dead Animals Based on the Amount
50.1 and Distribution of Gas Observed on Diagnostic Imaging
DV, Dorsal-ventral; GE, gas embolism; HBOT, hyperbaric oxygen treatment; LL, lateral-lateral.
gradually decreased over a period of 4 hours to a final can be placed in a plastic film–sealed tub connected to an
pressure of 1.9 ATA, where the pressure is held constant oxygen concentrator partially filled with water up to the
for the next 6 hours. Then the pressure is slowly further level of the nares of the turtle to help improve breathing
reduced back to atmospheric pressure (1 ATA) over a period (see Fig. 50.4D).
of 2 hours. It is essential to continuously observe breathing In the authors’ experience with more than 130 bycaught
during the entire procedure, especially during periods when loggerheads, treatment success mostly depends on severity
the pressure decreases, because breath holding during this of clinical signs at arrival, total amount and distribution of
time can lead to barotrauma. The current protocol is still intravascular gas, time to HBOT, and evidence of pulmo-
experimental and can be modified according to different nary water aspiration.
technical requirements. To maximize safety, it is important Sublethal effects cannot be ruled out in affected surviving
to remember that oxygen is flammable and explosive. individuals. In fact, cardiac, renal, and CNS damage have
If there is not access to a hyperbaric chamber, the patient been evident on histopathology in some individuals that
will benefit from breathing normobaric oxygen. Turtles survived the initial insult.
CHAPTER 50 Decompression Medicine in Aquatic Species (Fish and Sea Turtle Focus) 351
A B
C D
• Figure 50.4 (A) Computed tomography (CT) midsagittal image from a loggerhead sea turtle (Caretta
caretta) suffering from mild gas embolism. Notice that even in mild cases, abnormal gas is detected in
spinal cord region (asterisk). (B) CT transverse image of middle-coelom from severe gas embolism. Image
reveals severe presence of intravascular gas throughout hepatic parenchyma and enlarged liver volume.
Lungs appear dorsally compressed by liver expansion. (C) Custom-made hyperbaric chamber for sea
turtle hyperbaric oxygen therapy. (D) Container for alternative treatment with normobaric oxygen by means
of a hermetic plastic film sealed basin connected to an oxygen concentrator.
coming out of solution in the bloodstream and forming This technique has been widely promoted because of its
emboli in different tissues, and in fish most commonly ease of use for mitigation of acute barotrauma. However,
around the eyes, subcutaneous spaces under skin and the efficacy has been under intense debate and there are
flippers, or in the gill filaments. This condition is most contradictory results from different studies. Some studies
commonly known as gas bubble disease (GBD) syndrome indicate that venting the swim bladder at the surface with a
in aquarium fishes, with cold-water fishes being especially syringe and repositioning the stomach when everted, results
sensitive. Nitrogen is the main problematic gas because in only approximately 40%–50% survival of the fish at
oxygen is assimilated metabolically and thus is less likely to the surface and an unknown mortality following release.
form persistent bubbles.20 Regardless of gas bubble origin Increased survival has been reported by placing scuba divers
(decompression or supersaturation), treatment approach at mid depth to vent air bladders of fish coming up a
for excess of entrapped gas and/or gas bubble formation line from depth.24,25 Some authors point out that there is
can overlap for both processes, with focus on eliminating little evidence that venting increases survival, being equally
retained pathologic gas/bubbles. ineffective for freshwater and marine fishes. In other studies
the effects of venting varied with capture depth, with the
Diagnosis efficacy decreasing with depth.26 The available evidence
suggests that venting is generally discouraged in favor of
Observable signs of barotrauma in fish may include stomach recompression therapy. However, some studies still consider
eversion due to swim bladder overexpansion or even rupture, venting as an alternative option for barotrauma treatment
exophthalmia caused by retrobulbar gas emboli, intraocular in some fish species, reaching very high post reintroduc-
gas bubble formation potentially visible in the cornea and/ tion survivorship comparable to recompression therapy if
or anterior chamber of the eye, and emphysema in dermis applied rapidly enough.27 Following venting, an injured
especially visible in the fins (Fig. 50.5A, B, and E).1 Patients swim bladder may not be capable of regulating volume
are generally positively buoyant and in most cases unable to properly, or at best it may require a prolonged recovery
swim back to depth without intervention. Depending on period; so although venting can be practical under certain
severity, GE can be detected through diagnostic imaging circumstances and conditions where recompression is not
techniques, mostly ultrasound (see Fig. 50.5D), radiogra- feasible, the recommendation at present would be to apply
phy (see Fig. 50.5C), and CT; or bubbles can be seen in full recompression instead of venting, whenever possible.
blood vessels of virtually any organ, including skin (see Fig. For gas exophthalmia, mechanical removal of the gas
50.5B), gills, eyes (see Fig. 50.5A), viscera, and peritoneal from the retrobulbar space or even from inside the eye
cavity during necropsy. Common findings include vascular can be achieved using the appropriate syringe and needle.
and cardiac air, swim bladder overdistension or rupture, The needle should be the smallest gauge possible and long
hematomas, hemorrhages, and potentially internal organ enough to reach the gas pocket, which is easily identifiable
torsion associated with displacement.21,22 through radiography or guided by ultrasound (see Fig.
50.5C and D). This intervention should be repeated as
Treatment required throughout the course of the disease. Ultrasound
would typically aid in verifying the gas location within the
Although prevention is always a better approach than eye as well as potential damaged eye structures before and
treatment when dealing with decompressed fish, affected after treatment. If gas bubbles in the eye are left untreated,
patients once diagnosed should be recompressed as soon the consequences for the fish could include buphthalmos,
as possible to minimize tissue damage. Treatment has the rupture of the globe, atrophy (phthisis bulbi), infection,
traditionally been addressed by two means: venting or and even death.
recompression. Hyperbaric treatment is another treatment option.
Venting involves the insertion of a hypodermic needle Different studies reference that rockfishes showing severe
through the body wall to deflate a fish’s swim bladder or barotrauma can recover if quickly returned back to depth21,28
any air cavity. Major complications of barotrauma can be or with recompression in hyperbaric chambers.23,29 Time
quickly corrected with relative ease through the use of to recompression seems to be critical, and in some cases,
venting, but the outcome varies depending on the applica- animals can develop temporary or even permanent lesions
tion, fish species, and severity of trauma.23 associated with original barotrauma and GE.
• Figure 50.5 (A) John Dory fish (Zeus faver) showing intraocular bubbles. (B) Subcutaneous bubbles in dorsal fin from John Dory fish. (C) Head
dorsal-ventral radiographic comparison from juvenile piper gurnard fish (Trigla lyra) with retrobulbar gas compared with normal. Note radiolucent
anomaly behind the eye and the exophthalmia on the right x-ray. (D) Ocular ultrasound from two Ballan wrasse (Labrus bergylta). Upper image
shows normal anatomy with posterior echo reinforcement. Bottom study revealed marked exophthalmia and comet tail artifact behind the eye due
to presence of gas. (E) Rosy rockfish (Sebastes rosaceus) showing bilateral exophthalmia and corneal gas bubbles, as well as tight body (by swim
bladder distension) due to decompression event after being caught around 100 m deep. (Courtesy Bonnie Rogers, California State University).
(F) PVC double hyperbaric chamber used by Monterey Bay Aquarium for fish recompression treatment (Courtesy Joe Welsh, Monterey Bay
Aquarium).
CHAPTER 50 Decompression Medicine in Aquatic Species (Fish and Sea Turtle Focus) 353
A B
C D
E F
354 SE C T I O N 10 Aquatic
Recompression treatment can be done by two different system contains bypass valves to allow upper chamber
means: resubmersion of the fish to depth or the use of a pressurize/depressurize cycles, water source changes, or
hyperbaric chamber. other flow adjustments without disturbing a steady pres-
Resubmersion of fish to depth with remote sinking release sure in the main chamber. Because this second model is
devices or adapted cages: bottom cage/trap in the tank/ filled with pressurized water instead of gas, the risk for
ocean major accidents is less. For safety reasons, these chambers
One study showed that rockfish of the genus Sebastes made of PVC piping should not be used with pressurized
sp. caught at depths near 100 m, and then decompressed gases and especially oxygen.
but rapidly returned to depth and released, survived several Both single and double hyperbaric chambers are portable,
months developing active healthy reproductive organs.30 inexpensive, and work well for fish up to 40 cm.23
The California Department of Fish and Game, in collabo- Pressure in hyperbaric chambers can be adjusted to suit
ration with California and Oregon Sea Grant, has published the patient, depending on severity of signs, symptoms, and
a brochure called “Bring That Rockfish Down” (2008), in the kind of chamber available. The animals should be rapidly
which they promote and suggest methods for fishers to pressurized until normal buoyancy is restored or they sink
sink unwanted or prohibited species back to depth. This due to swim bladder gas compression. Fish are generally
brochure also recommends against the venting or gas aspira- initially recompressed at 5 ATM (6 ATA) for approximately
tion technique. The National Oceanic and Atmospheric 24 hours and then a controlled decompression (0.2–0.7
Administration has held barotrauma workshops and has ATM) every few hours (typically in periods over 8–10
opened a website (www.fishsmart.org) also encouraging the hours). The whole process can take from 3–4 days to more
use of sinking release devices. These devices range from the than a week.23 Fish buoyancy should be controlled during
simple and homemade to commercially available remotely the whole process. In case uncontrolled positive buoyancy
operated or pressure-activated units.23 is observed, recompression should be reapplied and pressure
In aquariums, it is not uncommon to put positively reduction should be more gradual. If fish are still slightly
buoyant or gas bubble–affected individuals on a cage at overbuoyant when taken out of the chamber, a cage can be
the bottom of a deep tank to facilitate gas reabsorption. used to retain the fish for a few more hours at the bottom
The trap should be lined with fine mesh synthetic netting of the tank until reaching neutral buoyancy.
to cushion the interior and avoid abrasion. This system Hyperbaric treatment is reported to be faster and with
of the cage and a weight can also be used in the ocean to decreased probability of relapse than when using the gas
send back to depth fished dysbaric individuals, increasing aspiration technique or venting. In addition, the prognosis
survivorship. is generally better.
Hyperbaric chambers have been used successfully to
recuperate fish suffering from decompression injury when Supportive Medical Treatment
collected at depth, as well as to treat fish suffering with
GBD, including gas exophthalmia or gas bubble pockets Combination of injectable antibiotics, antiinflammatories,
under skin. and a carbonic anhydrase inhibitor (acetazolamide) are
In addition, the Monterey Bay Aquarium (MBA) is one commonly used as required (there are frequent relapses)
of the most experienced institutions using two types of on GE/GBD-affected fishes to facilitate recovery until no
hyperbaric chambers for treatment of fish: pathologic gas pockets are present.
1. Single Chamber: a single chamber made of stainless steel Present studies indicate physical injuries, and behavioral
filled to approximately two-thirds capacity with seawater impairment associated with dysbarism may compromise
and then pressurized with oxygen while placed into a fish in the hours and weeks following onset, even with
refrigerated chamber or on ice to help reduce the water recompression. Potential visual aftereffects in fish that had
temperature and increase the gas solubility of the water. experienced exophthalmia, buphthalmia, and/or intraocular
Depending on the number of fish in the chamber, the gas are of concern. There is evidence of optic nerve stretch-
chamber can remain pressurized up to 8 hours before gas/ ing and visual impairment associated with dysbaric exoph-
water renovation is needed. Hyperbaric oxygen toxicity, thalmia.32,33 In case of gas exophthalmia, prognosis is better
although reported for some vertebrate species with pres- if the gas is located in the retrobulbar space compared with
sures over 2.8–3.0 ATA,31 does not seem to be affecting gas bubbles located inside the globe; as in the latter, bubbles
fish treated in these pressure chambers. The authors have can cause significant damage to eye structures, including
used pressurized oxygen at 4 ATA without any evident retinal detachment and complete vision loss.
deleterious effect in fish, whereas the MBA reports that
their standard oxygen pressurization is 7 ATA for 4–8 Acknowledgments
hours, and they never observed any problem associated
to it. We are grateful to the many excellent colleagues, animal
2. Double chamber: a custom-made chamber of polyvinyl care specialists, technical personnel from local authorities,
chloride (PVC) pipe that includes a staging chamber, being and students who have enabled us to complete part of
pressurized with a water pump. This double-chambered the studies mentioned in this chapter. Many thanks to all
CHAPTER 50 Decompression Medicine in Aquatic Species (Fish and Sea Turtle Focus) 355
professionals who have inspired us and supported our work 15. Norton TM, editor: Chelonian emergency and critical care. Semi-
along these years. Our special gratitude to two main referees nars in avian and exotic pet medicine, 2005, Elsevier.
of this work, Dr. Andreas Fahlman and Dr. Sally Nofs, 16. Litscher G: Ten years evidence-based high-tech acupuncture part
3: a short review of animal experiments, Evid Based Complement
who provided constructive criticism to the content of this
Alternat Med 7(2):151–155, 2010.
chapter, improving it significantly. Finally our thanks to Mr. 17. Hannah RW, Rankin PS, Penny AN, et al: Physical model of the
Joe Welsh from MBA, who provided additional comments development of external signs of barotrauma in Pacific rockfish,
and references for discussion, as well as a great number of Aquatic Biol 3(3):291–296, 2008.
images to be included in this chapter. 18. Hauck A: Gas bubble disease due to helicopter transport of
young pink salmon, Trans Amer Fish Soc 115(4):630–635, 1986.
References 19. Noga EJ: Gas supersaturation. In Noga EJ, editor: Fish disease:
diagnosis and treatment, 2011, John Wiley & Sons, pp 107–109.
1. Brown RS, Pflugrath BD, Colotelo AH, et al: Pathways of baro- 20. Marking LL: Gas supersaturation in fisheries: causes, concerns,
trauma in juvenile salmonids exposed to simulated hydroturbine and cures. DTIC Document, 1987.
passage: Boyle’s law vs. Henry’s law, Fish Res 121:43–50, 2012. 21. Jarvis ET, Lowe CG: The effects of barotrauma on the catch-
2. Francis TJR, Mitchell SJ: Manifestations of decompression and-release survival of southern California nearshore and shelf
disorders. In Brubakk AO, Neuman TS, editors: Bennett and rockfish (Scorpaenidae, Sebastes spp.), Can J Fish Aquat Sci
Elliott’s physiology and medicine of diving, 2003, Saunders, pp 65(7):1286–1296, 2008.
578–599. 22. Pribyl AL, Kent ML, Parker SJ, et al: The response to forced
3. Vann RD, Butler FK, Mitchell SJ, et al: Decompression illness, decompression in six species of Pacific rockfish, Trans Amer Fish
Lancet 377(9760):153–164, 2011. Soc 140(2):374–383, 2011.
4. Mahon RT, Regis DP: Decompression and decompression sick- 23. Welsh J: Hyperbaric chambers for fish, Diving for science
ness, Compr Physiol 4(3):1157–1175, 2014. 2012:129.
5. Gerth WA, Ruterbusch VL, Long ET: The influence of thermal 24. Starr RM, Heine JN, Johnson KA: Techniques for tagging and
exposure on diver susceptibility to decompression sickness. tracking deepwater rockfishes, N Am J Fish Manag 20(3):597–609,
Panama City, FL: NavSea, November 2007. Report No.: Con- 2000.
tract No.: NEDU TR 06-07. 25. Parrish FA, Moffitt RB: Subsurface fish handling to limit decom-
6. Fahlman A: Allometric scaling of decompression sickness risk in pression effects on deepwater species, Mar Fish Rev 54(3):29–32,
terrestrial mammals; cardiac output explains risk of decompres- 1992.
sion sickness, Sci Rep 7(40918):1–9, 2017. 26. Wilde GR: Does venting promote survival of released fish?
7. Lillo RS, Himm JF, Weathersby PK, et al: Using animal data Fisheries 34(1):20–28, 2009.
to improve prediction of human decompression risk following 27. Drumhiller KL, Johnson MW, Diamond SL, et al: Venting or
air-saturation dives, J Appl Physiol 93(1):216–226, 2002. rapid recompression increase survival and improve recovery of
8. Garcia-Parraga D, Crespo-Picazo JL, de Quiros YB, et al: Red Snapper with barotrauma, Mar Coast Fish 6(1):190–199,
Decompression sickness (‘the bends’) in sea turtles, Dis Aquat 2014.
Organ 111(3):191–205, 2014. 28. Hannah RW, Matteson KM: Behavior of nine species of Pacific
9. Fahlman A, Crespo-Picazo JL, Sterba-Boatwright B, et al: Defin- rockfish after hook-and-line capture, recompression, and release,
ing risk variables causing gas embolism in loggerhead sea turtles Trans Amer Fish Soc 136(1):24–33, 2007.
(Caretta caretta) caught in trawls and gillnets, Sci Rep 2017. In 29. Smiley JE, Drawbridge MA: Techniques for live capture of deep-
press. water fishes with special emphasis on the design and application
10. Stabenau EK, Vietti K: The physiological effects of multiple of a low-cost hyperbaric chamber, J Fish Biol 70(3):867–878,
forced submergences in loggerhead sea turtles (Caretta caretta), 2007.
Fish Bull 101(4):889–899, 2003. 30. Blain BJ, Sutton TM: Reproductive status and blood plasma
11. Harms CA, Mallo KM, Ross PM, et al: Venous blood gases and indicators of sex and gonad maturation status for yelloweye
lactates of wild loggerhead sea turtles (Caretta caretta) following rockfish following barotrauma and recompression events, Trans
two capture techniques, J Wildl Dis 39(2):366–374, 2003. Amer Fish Soc 145(6):1234–1240, 2016.
12. Snoddy JE, Williard AS: Movements and post-release mortality 31. Barthelemy L, Belaud A, Chastel C: A comparative study of
of juvenile sea turtles released from gillnets in the lower Cape oxygen toxicity in vertebrates, Respir Physiol 44(2):261–268,
Fear River, NC, USA, Endanger Species Res 12(3):235–247, 1981.
2010. 32. Rogers BL, Lowe CG, Fernández-Juricic E: Recovery of visual
13. De Quirós YB, González-Díaz Ó, Saavedra P, et al: Methodology performance in rosy rockfish (Sebastes rosaceus) following
for in situ gas sampling, transport and laboratory analysis of gases exophthalmia resulting from barotrauma, Fish Res 112(1):1–7,
from stranded cetaceans, Sci Rep 1:193, 2011. 2011.
14. De Quirós YB, González-Díaz O, Møllerløkken A, et al: 33. Rankin PS, Hannah RW, Blume MT, et al: Delayed effects of
Differentiation at autopsy between in vivo gas embolism and capture-induced barotrauma on physical condition and behav-
putrefaction using gas composition analysis, Int J Legal Med ioral competency of recompressed yelloweye rockfish (Sebastes
127(2):437–445, 2013. ruberrimus), Fish Res 186:258–268, 2017.
SECTION 11
56 Ophidiomycosis, 394
356
51
Euthanasia of Ectotherms
GREGORY A. LEWBART
357
358 SE C T I O N 11 Amphibians and Reptiles
TABLE
51.1 Euthanasia Methods for Invertebrates
Dosage/
Invertebrate Compound/Method Concentration Comments References
Sponges (Porifera)
Magnesium chloride 7.5% This is a two-step euthanasia* 34, 35
Coelenterates
Jellyfish Eugenol 120 mg/L Moon jellies (Aurelia aurita). 36
Two-step euthanasia*
MS-222 (tricaine 300–600 mg/L Moon jellies (Aurelia aurita). 37, 38
methanesulfonate); Two-step euthanasia*
buffered
Gastropods
Terrestrial Snails Ethanol 5% Two-step euthanasia* 2
““ Decarbonated beer 4.74% Two-step euthanasia* 2
Aquatic snails Ethanol/menthol (Listerine) 10% in saline Two-step euthanasia* 39
Magnesium salts (magnesium 20%–30% Two-step euthanasia* 34, 40
chloride or magnesium
sulfate)
Aplysia sp. (sea Magnesium chloride or 7.5% intracoelomic Two-step euthanasia* 41
hares) magnesium sulfate injection
Sodium pentobarbital 0.4 mg/mL H2O Two-step euthanasia* 42
Abalone 2-phenoxyethanol 0.5–3 mL/L Two-step euthanasia* 43
Benzocaine 100 mg/L Two-step euthanasia* 44
Magnesium sulfate 4–22 g/100 mL water Two-step euthanasia* 43
Sodium pentobarbital 1 mL/L (400 mg/L) Two-step euthanasia* 42, 45, 46
2-phenoxyethanol 0.5–3.0 mL/L * 41
Cephalopods
Ethanol 3%–10% in seawater Two-step euthanasia* 11, 47
Magnesium chloride 7%–10% in seawater Two-step euthanasia* 11, 47
Bivalves
Oysters
Magnesium chloride (3.5%) Variable effects with long 48–50
induction. Two-step euthanasia*
Propylene phenoxetol 1–3 mL/L Dosage is species dependent. 49, 50
(1% solution) Has been used in Tridacna
clams. Two-step euthanasia*
Scallops
Chloral hydrate 4 g/L Variable effects, temperature 51
dependent*
Magnesium chloride 30–50 g/L immersion Two-step euthanasia* 51
Annelida
Leeches Benzocaine 400 mg/L Hirudo medicinalis. May be 52
suitable for other aquatic
annelids*
Ethanol 5% Hirudo medicinalis. As an 53, 54
immersion*
Earthworms Ethanol 5% Lumbricus terrestris. As an 55
immersion*
CHAPTER 51 Euthanasia of Ectotherms 359
TABLE
51.1 Euthanasia Methods for Invertebrates—cont’d
Dosage/
Invertebrate Compound/Method Concentration Comments References
Arachnida
Tarantulas Alfaxalone 200 mg/kg intracardiac Combine with: 20 mg/kg 56
ketamine, 5.0 mg/kg morphine,
or 20 mg/kg xylazine*
Spiders and Carbon dioxide 3%–5% Induction chamber system* 35
Scorpions Isoflurane/Sevoflurane 3%–5%/4%–6% Induction chamber system* 53, 57–59
Crustaceans
Small crabs; Eugenol (clove oil) 0.125 mL/L * 60, 61
Lobsters 75–100 ppm
Hermit crabs Isoflurane 59
Marine crabs Ketamine 20 mg/kg IV Reliable and deep anesthesia 62
Add xylazine (20 mg/kg) for
greater effects*
Crayfish Ketamine 40–90 mg/kg IM * 63
Lidocaine 400–1000 mg/kg IM * 63
American Potassium chloride 1 g/kg (330 mg/mL Inject at base of second walking 64
lobsters solution) IV leg
Crabs Procaine 25 mg/kg IV 65
Xylazine 16–22 mg/kg IV * 60
Echinoderms
Sea stars Magnesium chloride 7.5%–8% * 66, 67
MS-222 (tricaine 1–10 g/L * 68
methanesulfonate);
buffered
Sea urchins MS-222; buffered 0.4–0.8 g/L * 69
*A two-step euthanasia is defined as euthanasia that requires a second method to humanely kill the animal. In most cases the animal has been rendered
unconscious by the first step before the second step, which may include a fixative such as 10% neutral buffered formalin or 70% ethanol, is applied. Freezing
or physical destruction may also constitute second-step applications.
are currently more than 7600 described species, almost Table 51.3 summarizes the commonly utilized methods,
90% of which are anurans (frogs and toads).26 The other both pharmaceutical and physical, with comments and
two orders, Caudata (salamanders) and Gymnophiona references where appropriate.
(caecilians), contain 9% and 3% of the species, respectively.
Most species have a two-stage life cycle and can respire Reptiles
through their skin. This presents different challenges when
euthanasia is being considered. For example, euthanasia for The class Reptilia is broken into 4 orders and 48 families
a larval form (tadpole) might require a different protocol containing 10,450 species, as of August 2016.30 Taxonomy
than a metamorphosed individual (frog or salamander). and nomenclature are dynamic disciplines, so these numbers
Lillywhite et al. provide a detailed review of anesthesia change frequently and may not be universally accepted by
and euthanasia in amphibians.27 Other recent articles that herpetologists. The snakes and the lizards belong in the
address these topics include Atwood28 and Shine et al.29 same order (Squamata) and the turtles, crocodilians, and the
360 SE C T I O N 11 Amphibians and Reptiles
TABLE
51.2 Euthanasia Methods for Fishes
*A two-step euthanasia is defined as euthanasia that requires a second method to humanely kill the animal. In most cases the animal has been rendered
unconscious by the first step before the second step, which may include double pithing, decapitation, or injectable pentobarbital.
TABLE
51.3 Euthanasia Methods for Amphibians
*A two-step euthanasia is defined as euthanasia that requires a second method to humanely kill the animal. In most cases the animal has been rendered
unconscious by the first step before the second step, which may include double pithing, decapitation, or injectable pentobarbital.
TABLE
51.4 Euthanasia Methods for Reptiles
*A two-step euthanasia is defined as euthanasia that requires a second method to humanely kill the animal. In most cases the animal has been rendered
unconscious by the first step before the second step, which may include double pithing, decapitation, or injectable pentobarbital.
362 SE C T I O N 11 Amphibians and Reptiles
References 24. Valentim AM, van Eeden FJ, Strähle U, et al: Euthanizing
zebrafish legally in Europe; Are the approved methods of
1. Leary S, Underwood W, Anthony R, et al: AVMA Guidelines for euthanizing zebrafish appropriate to research reality and animal
the euthanasia of animals: 2013 edition, AVMA, 2013, Schaum- welfare?, EMBO Rep 17(2):1688–1689, 2016.
burg, IL. 25. Lidster K, Readman GD, Prescott MJ, et al: International survey
2. Gilbertson CR, Wyatt JD: Evaluation of euthanasia techniques on the use and welfare of zebrafish Danio rerio in research, J Fish
for an invertebrate species, land snails (Succinea putris), J Am Biol 2017, doi:10.1111/jfb.13278.
Assoc Lab Anim Sci 55(5):577–581, 2016. 26. Amphibiaweb. Amphibiaweb.org. (Accessed 15 April 2017).
3. Murray MJ: Euthanasia. In Lewbart GA, editor: Invertebrate 27. Lillywhite HB, Shine R, Jacobson E, et al: Anesthesia and
medicine, ed 2, 2012, Wiley-Blackwell, pp 441–443. euthanasia of amphibians and reptiles used in scientific research:
4. Zachariah T: Invertebrate animal welfare. In Lewbart GA, Should hypothermia and freezing be prohibited?, Bioscience
editor: Invertebrate medicine, ed 2, 2012, Wiley-Blackwell, pp 67(1):53–61, 2017.
445–449. 28. Atwood MA: Effects of euthanasia method on stable-carbon and
5. Crook RJ: The welfare of invertebrate animals in research: Can stable-nitrogen isotope analysis for an ectothermic vertebrate,
science’s next generation improve their lot?, PostDoc J 1(2):9–20, Rapid Commun Mass Spectrom 28:909–913, 2013.
2013. 29. Shine R, Amiel J, Munn AJ, et al: Is cooling then freezing a
6. Horvath K, Angeletti D, Nascetti G, et al: Invertebrate welfare: humane way to kill amphibians and reptiles?, Biol Open 2013,
an overlooked issue, Ann Ist Super Sanitá 49(1):9–17, 2013. doi:10.1242/bio.012179.
7. Fiorito G, Affuso A, Basil J, et al: Guidelines for the care and 30. Reptile Database. Reptile-database.org/db-info/SpeciesStat.html.
welfare of cephalopods in research – A consensus based on an (Accessed 15 April 2017).
initiative by CephRes, FELASA and the Boyd Group, Lab Anim 31. Nevarez JG, Strain GM, da Cunha AF, et al: Evaluation of four
49(S2):1–90, 2015. methods for inducing death during slaughter of American alliga-
8. Berry A, Vitale A, Carere C, et al: EU guidelines for the care and tors (Alligator mississippiensis), Am J Vet Res 75:536–543, 2014.
welfare of an “exceptional invertebrate class” in scientific research, 32. Mader DR: Euthanasia. In Mader DR, Divers SJ, editors:
Ann Ist Super Sanitá 51(4):267–269, 2015. Reptile medicine and surgery, ed 2, St. Louis, MO, 2006, Elsevier,
9. Cooke GM, Tonkins BM: Behavioural indicators of welfare pp 564–568.
exhibited by the common European cuttlefish (Sepia officinalis), 33. Music MK, Strunk A: Reptile critical care and common emergen-
J Zoo Aquar Res 3(4):157–162, 2015. cies, Vet Clin Exot Anim 19:591–612, 2016.
10. Mather JA, Carere C: Cephalopods are best candidates for 34. Brusca RC: Common intertidal invertebrates of the Gulf of Cali-
invertebrate consciousness, Animal Sentience 107, 2016. fornia, ed 2, Tucson, AZ, 1980, University of Arizona Press.
11. Gleadall IG: The effects of prospective anaesthetic substances 35. Gunkel C, Lewbart GA: Anesthesia and analgesia of invertebrates.
on cephalopods: Summary of original data and a brief review In Fish R, Danneman P, Brown M, et al, editors: Anesthesia
of studies over the last two decades, J Exp Mar Biol 447:23–30, and analgesia in laboratory animals, ed 2, St. Louis, MO, 2008,
2013. Elsevier, pp 535–546.
12. Andrews PLR, Darmaillacq A-S, Dennison N, et al: The iden- 36. Freeman KS, Lewbart GA, Robarge WP, et al: Characterizing
tification and management of pain, suffering, and distress in eversion syndrome in captive scyphomedusa jellyfish, Am J Vet
cephalopods, including anaesthesia, analgesia and human killing, Res 70(9):1087–1093, 2009.
J Exp Mar Biol and Ecol 447:46–64, 2013. 37. Stoskopf MK: Coelenterates. In Lewbart GA, editor: Invertebrate
13. Fishbase. Fishbase.org. (Accessed 15 April 2017). medicine, ed 2, 2012, Wiley-Blackwell Publishing, pp 21–56.
14. Sneddon LU: Pain perception in fish; indicators and endpoints, 38. Dombrowski D: Unpublished work. 2017.
ILAR J 50:338–342, 2009. 39. Woodall AJ, Naruo H, Prince DJ, et al: Anesthetic treatment
15. Sneddon LU: Pain perception in fish: evidence and implications blocks synaptogenesis but not neuronal regeneration of cultured
for the use of fish, J Conscious Stud 18:209–229, 2011. Lymnaea neurons, J Neurophysiol 90:2232–2239, 2003.
16. Sneddon LU: Pain in aquatic animals, J Exp Biol 218:967–976, 40. Messenger JB, Nixon M, Ryan KP: Magnesium chloride as an
2015. anaesthetic for cephalopods, Comp Biochem Physiol 82C:203–205,
17. Rose JD: The neurobehavioral nature of fishes and the question 1985.
of awareness and pain, Rev Fish Sci 10:1–38, 2002. 41. Clark TR, Nossov PC, Apland JP, et al: Anesthetic agents for use
18. Rose JD, Arlinghaus R, Cooke SJ, et al: Can fish really feel pain?, in the invertebrate sea snail, Aplysia californica, Contemp Top Lab
Fish Fisheries 15:97–133, 2014. Anim Sci 35(5):75–79, 1996.
19. Neiffer D, Stamper MA: Fish sedation, anesthesia, analgesia, and 42. Martins-Sousa RL, Negrao-Correa D, Bezerra FSM, et al:
euthanasia: Considerations, methods, and types of drugs, ILAR J Anesthesia of Biomphalaria spp. (Mollusca, Gastropoda): sodium
50:343–360, 2009. pentobarbital is the drug of choice, Mem Inst Oswaldo Cruz
20. Blessing JJ, Marshal JC, Balcombe SR: Humane killing of fishes 96:391–392, 2001.
for scientific research: a comparison of two methods, J Fish Biol 43. White HI, Hecht T, Potgieter B: The effect of four anaesthetics
76:2571–2577, 2010. on Haliotis midae and their suitability for application in com-
21. Balko JA, Oda A, Posner LP: Immersion euthanasia of goldfish mercial abalone culture, Aquaculture 140:145–151, 1996.
(Carassius auratus), Proc Inter Assoc Aquat Anim Med 2016. 44. Edwards S, Burke C, Hindrum S, et al: Recovery and growth
22. Bartlett DH, Silk SB: Office of laboratory animal welfare com- effects of anaesthetic and mechanical removal on greenlip (Hali-
ments, Zebrafish 13(6):2016, doi:10.1089/zeb.2016.1344. otis laevigata) and blacklip (Haliotis rubra) abalone, J Shellfish Res
23. Hawkins P, Prescott MJ, Carbone L, et al: A good death? Report 19(1):510, 2000.
of the second Newcastle meeting on laboratory animal euthana- 45. Aquilina B, Roberts R: A method for inducing muscle relaxation
sia, Animals 6(50):2016, doi:10.3390/ani6090050. in the abalone, Haliotis iris, Aquaculture 190:403–408, 2000.
CHAPTER 51 Euthanasia of Ectotherms 363
46. Sharma PD, Nollens HH, Keogh JA, et al: Sodium 62. Quesada RJ, Smith CD, Heard DJ: Evaluation of parenteral
pentobarbitone-induced relaxation in the abalone Haliotis iris drugs for anesthesia in the blue crab (Callinectes sapidus), J Zoo
(Gastropoda): effects of animal size and exposure time, Aquacul- Wild Med 42(2):295–299, 2011.
ture 218:589–599, 2003. 63. Brown PB, White MR, Chaille J, et al: Evaluation of three
47. Scimeca J: Cephalopods. In Lewbart GA, editor: Invertebrate anesthetic agents for crayfish (Orconectes virilis), J Shellfish Res
medicine, ed 2, Ames, IA, 2012, Wiley-Blackwell Publishing, pp 15:433–435, 1996.
113–125. 64. Battison A, MacMillans R, MacKenzie A, et al: Use of injectable
48. Culloty SC, Mulcahy MF: An evaluation of anesthetics for Ostrea potassium chloride for euthanasia of American lobsters (Homarus
edulis (L, Aquaculture 107(2–3):249–252, 1992. americanus), Comp Med 50:545–550, 2000.
49. Norton JH, Dashorst M, Lansky TM, et al: An evaluation of 65. Oswald RL: Immobilization of decapod crustaceans for experi-
some relaxants for use with pearl oysters, Aquaculture 144:39–52, mental purposes, J Mar Biol Assoc UK 57:715–721, 1977.
1996. 66. McCurley RS, Kier WM: The functional morphology of starfish
50. Mills D, Tlili A, Norton J: Large-scale anesthesia of the tube feet: The role of a crossed-fiber helical array in movement,
silver-lip pearl oyster, Pinctada maxima Jameson, J Shellfish Res Biol Bull 188:197–209, 1995.
16:573–574, 1997. 67. Harms CA: Echinoderms. In Lewbart GA, editor: Invertebrate
51. Heasman MP, O’Connor WA, Frazer AWJ: Induction of anesthe- medicine, Ames, IA, 2012, Wiley-Blackwell Publishing, pp
sia in the commercial scallop, Pecten fumatus Reeve, Aquaculture 365–379.
131:231–238, 1995. 68. O’Neill PL: The effect of anesthesia on spontaneous contraction
52. Cooper JE, Mahaffey P, Applebee K: Anaesthesia of the medicinal of the body wall musculature in the astereroid Coscinasterias
leech (Hirudo medicinalis), Vet Rec 118:589–590, 1986. calamaria, Mar Behav Physiol 24:137–150, 1994.
53. Cooper JE: Invertebrate anesthesia, Vet Clin North Am Exotic 69. Applegate JR, Dombrowski D, Christian LS, et al: Tricaine
Anim Pract 4:57–67, 2001. methanesulfonate (MS-222) sedation and anesthesia in the
54. Marks DH, Cooper EL: Aeromonas hydrophila in the coelomic purple-spined sea urchin (Arbacia punctulata), J Zoo Wild Med
cavity of the earthworms Lumbricus terrestris and Eisenia foetida, 47(4):1025–1033, 2016.
J Invert Pathol 29:382–383, 1977. 70. Noga EJ: Fish disease: Diagnosis and treatment, ed 2, Ames, IA,
55. Salgado MA, Lewbart GA, Christian LS, et al: Evalua- 2010, Wiley-Blackwell, p 519.
tion of five different suture materials in the skin of the 71. Wilson JM, Bunte RM, Carty AJ: Evaluation of rapid cooling
earthworm (Lumbricus terrestris), Springerplus 3:423, 2014, and tricaine methanesulfonate (MS222) as methods of euthana-
doi:10.1186/2193-1801-3-423. sia in zebrafish (Danio rerio), J Am Assoc Lab Anim 48:785–789,
56. Gjeltema J, Posner LP, Stoskopf MK: The use of injectable 2009.
alphaxalone as a single agent and in combination with ketamine, 72. Torreilles SL, McClure DE, Green SL: Evaluation and refine-
xylazine, and morphine in the Chilean rose tarantula, Gram- ment of euthanasia methods for Xenopus laevis, J Am Assoc Lab
mostola rosea, J Zoo Wild Med 45(4):792–801, 2014. Anim Sci 48(5):512–516, 2009.
57. Melidone R, Mayer J: How to build an invertebrate surgery 73. Burns RB, McMahan W: Euthanasia methods for ectotherm ver-
chamber, Exotic DVM 7(5):8–10, 2005. tebrates. In Bonagura JD, editor: Kirk’s current veterinary therapy
58. Pizzi R: Spiders. In Lewbart GA, editor: Invertebrate medicine, ed XII: Small animal practice, Philadelphia, PA, 1995, Saunders, pp
2, 2012, Wiley-Blackwell, pp 187–221. 1379–1381.
59. Marnell C: Tarantula and hermit crab emergency care, Vet Clin 74. von Esse FV, Wright KM: Effect of intracoelomic propofol in
Exot Anim Med 19:627–646, 2016. White’s tree frogs, Pelodryas caerulea, Bull Assoc Rept Amph Vet
60. Gardner C: Options for immobilization and killing crabs, J 7–8, 1999.
Shellfish Res 116:219–224, 1997. 75. Conroy CJ, Papenfuss T, Parker J, et al: Use of tricaine meth-
61. Waterstrat PR, Pinkham L: Evaluation of eugenol as an anesthetic anesulfonate (MS222) for euthanasia of reptiles, J Am Assoc Lab
for the American lobster Homarus americanus, J World Aquacult Anim Sci 48(1):28–32, 2009.
Soc 36(3):420–424, 2005.
52
Ranaviral Disease in Reptiles
and Amphibians
MATTHEW C. ALLENDER
W
ildlife diseases have been on the rise across the Various ranaviruses account for epizootics, including
world.1 Although some of these diseases receive a FV3, Ambystoma tigrinum virus (ATV), soft-shelled turtle
great deal of attention, such as chytridiomycosis iridovirus (STIV), and Bohle iridovirus (BIV). FV3 is the
in worldwide amphibian declines, white-nose syndrome in only iridovirus identified in turtles in North America and
North American bats (see also Chapter 72), West Nile virus is also the most commonly reported iridovirus for anurans.
in birds, and chronic wasting disease in cervids,1–3 others ATV affects salamanders in the western United States,
are poorly understood and require further study (see also whereas BIV is the main ranavirus identified in Austra-
Chapter 39). Disease events may have a dramatic impact lia.10,11 Many manuscripts describe the detection of these
on local populations, becoming more critical as population ranaviruses using only polymerase chain reaction (PCR) of
size decreases.1 One such fatal disease in amphibians and the major capsid protein (MCP); thus terminology such as
reptiles results from viruses in the family Iridoviridae. Infec- FV3-like virus indicates that the sequence was homologous
tions and death have been reported in numerous species, to the type species but further characterization was not
contributing to population declines. performed. This has meant very little in a clinical setting
The family Iridoviridae consists of two subfamilies and because disease syndrome for FV3 and FV3-like diseases are
five genera (Table 52.1).4 They are large, icosahedral, DNA assumed to be synonymous; thus all mention of FV3-like is
viruses that may be found in an enveloped or nonenveloped considered to be FV3 in this chapter.
form. Diseases caused by viruses in the genus Ranavirus Ranaviruses have a wide species distribution, with
are a growing concern in free-ranging and captive amphib- significant differences in species susceptibility even within
ian and reptile populations.5 These viruses are found the same isolate.12,13 There are several species that appear
worldwide, cause significant morbidity and mortality in uniquely sensitive to these viruses, including tiger salaman-
affected species,5 and were placed on the World Organisa- ders (Ambystoma tigrinum) to ATV,11 wood frogs (Lithobates
tion for Animal Health (OIE) list of reportable diseases for sylvaticus) to FV3,12,14,15 and eastern box turtles (Terrapene
amphibians.6 Detailed information on taxonomy, molecular carolina carolina) to FV3.16 However, all anurans, caudates,
characteristics, surveillance techniques, and ecology is avail- and chelonians should be considered susceptible.
able elsewhere.7 Age class often aids in diagnosis in amphibian diseases.
Many reports indicate that larval amphibians in North
Geographic and Host Distribution America are more susceptible to mortality than adults,
whereas adult mortality is more commonly reported in
The discovery of frog virus 3 (FV3) was first reported in European amphibians.9,17,18 Adult chelonians have been
leopard frogs (Rana pipiens) being studied for Lucke renal more commonly reported to develop FV3-like infections
adenocarcinomas in the 1960s.8 Since that time, numerous than juveniles.16 However, there are fewer overall reports in
outbreaks have occurred in both free-ranging and captive reptiles with FV3, leading to the likelihood that adult rep-
herpetofauna across the world.5,9 Outbreaks have occurred tiles are more likely sampled and diagnosed than juveniles.
throughout the world, including North America (Arizona, In Australia, juveniles of two species of tortoise (Krefft river
Colorado, Florida, Georgia, Idaho, Kentucky, Maine, Mas- turtle [Emydura krefftii] and saw-shelled tortoise [Elseya lati-
sachusetts, Minnesota, New Hampshire, New York, North sternum]) were susceptible to BIV, whereas adult tortoises,
Carolina, North Dakota, Pennsylvania, Tennessee, Texas, juvenile crocodiles, and three species of snakes were not
Utah, Wyoming, Saskatchewan, Ontario), South America affected after experimental transmission.18 All age classes of
(Brazil, Uruguay, Venezuela), Europe (Croatia, Denmark, reptiles and amphibians should be considered susceptible to
Germany, Netherlands, Spain, United Kingdom), Asia this disease; however, juvenile/larval age classes may be at
(China, Japan), and Australia.5 increased risk in some species.
364
CHAPTER 52 Ranaviral Disease in Reptiles and Amphibians 365
TABLE
52.1 Taxonomy of Iridoviruses and Species Affected
A B
C D
• Figure 52.1 (A) Clinical signs of a larval wood frog (Lithobates sylvaticus) demonstrating hemorrhages
of the tail base; (B), Larval silvery salamander (Ambystoma platineum) with disseminated hemorrhages;
(C), An eastern box turtle (Terrapene carolina carolina) with dehydration and lethargy; (D), An eastern
box turtle with lethargy, ocular, and nasal discharge. (Photos A and B Courtesy Kelsey Low; C and D
Courtesy Matthew Allender.)
were seen only at 22°C, but not at 28°C; control turtles at neuroepithelium, nasal tissues, adipose, trachea, muscle,
both temperatures demonstrated only rare lethargy (12.5%) and osteoclasts.9,21,24,29,30,36
or leg swelling (25%).21
Pathologic findings in susceptible individuals are similar Diagnosis
between adults and juveniles of both amphibians and
reptiles.9,33 Infections lead to systemic organ failure due Several methods have been proposed for the diagnosis of
to cellular necrosis in the spleen, liver, kidney, and intes- ranaviruses, including histopathology, PCR, virus isola-
tines.9,33 Hemorrhagic syndromes are common in anurans tion, enzyme-linked immunosorbent assay (ELISA), elec-
in the United Kingdom34 and in salamanders in western tron microscopy, restriction fragment length polymorphism
North America35 and were observed in one experimentally (RFLP), and cytology.9,24,37
inoculated red-eared slider.20 Tiger salamanders have been Conventional PCR targeting the MCP is commonly
observed with polypoid lesions early in the course of the used to identify infections but is limited by the fact that
disease that progress to cover most of the body.36 There are it allows detection of only a 530-bp segment (or less) and
also nonspecific changes noted with ranavirus infections, does not confirm active infection. Diagnosis is most success-
such as lymphocytosis, lymphoid depletion, and vacuola- ful on fresh or frozen tissues, but a method for successful
tion of hepatocytes and renal tubular cells.33 Basophilic viral recovery from formalin-fixed tissues exists.38 Evaluation of
inclusions have been inconsistently observed in affected antemortem (toe clip) versus postmortem (liver) samples
tissues.20,24 When present, inclusions have been identified in anurans was evaluated for detection of FV3.39 The study
in erythrocytes, leukocytes, epithelial cells, meninges, gills, found that 88% of samples that were positive in liver
CHAPTER 52 Ranaviral Disease in Reptiles and Amphibians 367
samples were also positive in toe clips, leading the authors that are less specific should also be considered when
to conclude that both methods are effective for detection.39 attempting to determine the presence or absence of these
The specificity and sensitivity of PCR were also evaluated viruses. Clinical pathology is commonly used to character-
comparing postmortem (necropsy) and antemortum (tail ize the health status of an animal. Although not specific,
clips) in salamanders.40 The study demonstrated that tail it may be used with other diagnostic methods in a parallel
clips underestimate prevalence, although its agreement testing method to potentially increase the sensitivities of
increases as time after exposure increases.40 In reptiles, the assays. Intracytoplasmic inclusions were identified in
necropsy tissues are most commonly tested. Agreement of an eastern box turtle with natural infection30 but rarely in
necropsy tissues were compared with antemortem sampling red-eared sliders following experimental infection.21 This
and concluded whole blood and oral swabs were 100% same experimental challenge found a significant decrease
specific and 100% sensitive, whereas cloacal swabs were in total protein over time.50
100% specific and 83% sensitive if comparing with any
sample taken during the 30-day trial.21 However, experi- Treatment and Prevention
mental transmission in eastern box turtles demonstrated
whole blood detection spiked at 13 days and oral swabs not Therapeutic interventions in free-ranging settings are often
until 16 days.26 In the same study, whole blood detection impractical for combating disease outbreaks. However, in
ended by day 27, but oral swab detection persisted until captive animals or in those situations in which an endangered
day 55.26 Therefore it is recommended that a dual testing group of free-ranging animals is brought into captivity, an
strategy of both whole blood and oral swabs is needed for appropriate treatment protocol needs to be established.
greatest testing success. Acyclovir is a guanine analog antiviral drug.51 It is active
Quantitative PCR (qPCR) has been developed to detect against herpesviruses due to the presence of the thymidine
fewer viral copies than conventional PCR. Primers target kinase (TK) enzyme, which rapidly activates acyclovir to the
a segment of the ranavirus DNA polymerase gene.41 This monophosphate form.52 Several isolates of iridoviruses have
assay was validated in cell culture for several ranaviruses and been shown to have TK genes or functional TK enzymes.53,54
was found to be effective in detecting virus in fish tissues In vitro studies against an iridovirus using acyclovir indi-
experimentally infected with epizootic hematopoietic cated only a dose-dependent partial inhibition at 25 µg/
necrosis virus (EHNV)41 but has difficulty in differentiating mL.16 The half-life of acyclovir after a single oral valcyclovir
ranavirus species. A similar assay was developed based on dose in eastern box turtles is 14.1 hours, and that of mar-
a 56-bp segment of the MCP of FV3 and validated in ginated tortoises (Testudo marginata) given oral acyclovir is
Terrapene heart cell culture, eastern box turtle whole blood 8.8 hours.55,56 These results are slower than homeotherms,
extracts, and plasmid-spiked cell cultures.42 which could allow a reduction in dosing intervals. Famci-
Immunohistochemistry is useful in quickly confirming clovir at three doses was used during a ranavirus outbreak in
the presence of a pathogen in formalin-fixed tissue. An eastern box turtles in a zoo.57 There were no significant dif-
immunohistochemical (IHC) method has been developed ferences in survival based on dose, but other supportive care
using rabbit antiserum against purified virus to definitively measures were used and may have contributed to survival.
demonstrate systemic ranavirus infection in several tissues Besides pharmacologic treatment, other mechanisms to
from fish and amphibians.43,44 Immunohistochemistry also control FV3 include temperature alteration and disinfec-
has been developed in the United Kingdom that is capable tion. The effect of temperature on the pathogenesis of irido-
of detecting ranavirus in lymphocytes, fibrocytes, and mela- viruses has been shown to be important. Tiger salamanders
nomacrophages, among other tissues from amphibians.45 experimentally inoculated with ATV showed high mor-
IHC may be an invaluable diagnostic tool when working tality at 10°C and 18°C, whereas lower mortalities were
on an outbreak and has been used in Australia for this noted at 26°C.58 Furthermore, viral loads were highest in
purpose.46 However, IHC may be applied only to biopsy or the animals kept at 10°C and lowest in the animals kept
necropsy tissues, which are an invasive method that is not a at 26°C.58 In adult red-eared sliders, there was 100% mor-
preferred method for antemortem diagnosis. tality in turtles exposed to FV3 at 22°C and only 50% at
Serologic assays have been developed to detect antibodies 28°C.21 Although FV3 growth is inhibited at temperatures
against iridoviruses in amphibians and reptiles.18,47 Antibod- greater than 32°C, the effects of other ranaviruses in other
ies in cane toads (Bufo marinus) were detected using an species clearly indicate either virus or species-specific dif-
ELISA against purified BIV and EHNV.48,49 In chelonians, ferences in pathogenesis, and further studies are needed in
FV3 antigen and an anti–desert tortoise IgY were used several species to better characterize the role of temperature
in an indirect ELISA.47 This assay demonstrated a high in development of disease.
coefficient of variation (>15% in some replicates) but was Chlorhexidine at 0.75%, sodium hypochlorite at 3%,
used for evaluation of gopher tortoises and box turtles with and potassium peroxymonosulfate at 1% were all effective at
low prevalence (<1.5%)47; however, the high coefficient of inactivating ranavirus after a 1-minute exposure,59 whereas
variation may have led to false positives. potassium permanganate (100% concentration) after a
Although the previous diagnostic tests are specific to 60-minute exposure, chlorhexidine at 0.25%, and sodium
characterizing ranaviruses, additional diagnostic methods hypochlorite less than 3% were found to be ineffective.59
368 SE C T I O N 11 Amphibians and Reptiles
11. Greer AL, Brunner JL, Collins JP: Spatial and temporal patterns 29. Docherty DE, Meteyer CU, Wang J, et al: Diagnostic and
of Ambystoma tigrinum virus (ATV) prevalence in tiger sala- molecular evaluation of three iridovirus-associated salamander
manders Ambystoma tigrinum nebulosum, Dis Aquat Org 85:1–6, mortality events, J Wildl Dis 39:556–566, 2003.
2009. 30. Allender MC, Fry MM, Irizarry AR, et al: Intracytoplasmic
12. Schock DM, Bollinger TK, Chinchar VG, et al: Experimental inclusions in circulating leukocytes from an eastern box turtle
evidence that amphibian ranaviruses are multi-host pathogens, (Terrapene carolina carolina) with iridoviral infection, J Wildl Dis
Copeia 133–143, 2008. 42:677–684, 2006.
13. Hoverman JT, Gray MJ, Miller DL: Anuran susceptibilities to 31. De Voe RK, Geissler K, Elmore S, et al: Ranavirus-associated
ranaviruses: role of species identity, exposure route, and a novel morbidity and mortality in a group of captive eastern box turtles
virus isolate, Dis Aquat Org 89:97–107, 2010. (Terrapene carolina carolina), J Zoo Wildl Med 35:534–543,
14. Greer AL, Berrill M, Wilson PJ: Five amphibian mortality events 2004.
associated with ranavirus infection in south central Ontario, 32. Allender MC, Mitchell MA, McRuer D, et al: Prevalence, clini-
Canada, Dis Aquat Org 67:9–14, 2005. cal signs, and natural history characteristics of frog virus 3-like
15. Harp EM, Petranka JW: Ranavirus in wood frogs (Rana sylvatica): infections in eastern box turtles (Terrapene carolina carolina),
potential sources of transmission within and between ponds, J Herp Conserv Biol 8:308–320, 2013.
Wildl Dis 42:307–318, 2006. 33. Miller DL, Gray MJ, Rajeev S, et al: Pathologic findings in larval
16. Johnson AJ: Iridovirus infections of captive and free-ranging and juvenile anurans inhabiting farm ponds in Tennessee, USA,
chelonians in the United States, Gainesville, 2006, University of J Wildl Dis 45:314–324, 2009.
Florida, Veterinary Medicine. 34. Cunningham AA, Hyatt AD, Russell P, et al: Experimental trans-
17. Brunner JL, Storfer A, Gray MJ: Ranavirus ecology and evolution: mission of a ranavirus disease of common toads (Bufo bufo) to
from epidemiology to extinction. In Gray MJ, Chinchar VG, common frogs (Rana temporaria), Epidem Infect 135:1213–1216,
editors: Ranaviruses: Lethal pathogens of ectothermic vertebrates, 2007.
2015, Springer Open Publishing, pp 71–104. 35. Bollinger TK, Mao J, Schock D, et al: Pathology, isolation, and
18. Ariel E: Pathology and serological aspects of Bohle iridovirus preliminary molecular characterization of a novel iridovirus from
infections in six selected water-associated reptiles in North tiger salamanders in Saskatchewan, J Wildl Dis 35:413–429,
Queensland, 1997. North Queensland, James Cook University, 1999.
Department of Microbiology and Immunology. 36. Jancovich JK, Davidson EW, Morado JF, et al: Isolation
19. Johnson AJ, Pessier AP, Wellehan JFX, et al: Ranavirus infection of a lethal virus from the endangered tiger salamander
of free-ranging and captive box turtles and tortoises in the United Ambystoma tigrinum stebbinsi, Dis Aquat Org 31:161–167,
States, J Wildl Dis 44:851–863, 2008. 1997.
20. Johnson AJ: Experimental transmission and induction of 37. Miller DL, Pessier AP, Hick P, et al: Comparative pathology of
ranaviral disease in western ornate box turtles (Terrapene ornata ranaviruses and diagnostic techniques. In Gray MJ, Chinchar
ornata) and red-eared sliders (Trachemys scripta elegans), Vet Pathol VG, editors: Ranaviruses: lethal pathogens of ectothermic verte-
44:285–297, 2007. brates, 2015, Springer Open Publishing, pp 171–208.
21. Allender MC, Mitchell MA, Torres TA, et al: Pathogenecity 38. Kattenbelt JA, Hyatt AD, Gould AR: Recovery of ranavirus
of frog-virus 3-like virus in red-eared slider turtles (Trachemys dsDNA from formalin-fixed archival material, Dis Aquat Org
scripta elegans) at two environmental temperatures, J Comp Pathol 39:151–154, 2000.
149:356–367, 2013. 39. St-Amour V, Lesbarreres D: Genetic evidence of ranavirus in toe
22. Brenes R, Gray MJ, Waltzek TB, et al: Transmission of rana- clips: an alternative to lethal sampling methods, Conserv Genet
virus between ectothermic vertebrates, PLoS ONE 9:e92476, 8:1247–1250, 2007.
2014. 40. Greer AL, Collins JP: Sensitivity of a diagnostic test for amphibian
23. Kimble SJ, Karna AK, Johnson AJ, et al: Mosquitoes as a ranavirus varies with sampling protocol, J Wildl Dis 43:525–532,
potential vector of ranavirus transmission in terrestrial turtles, 2007.
Ecohealth 12:334–338, 2015. 41. Holopainen R, Honkanen J, Jensen BB, et al: Quantitation
24. Gray MJ, Miller DL, Hoverman JT: Ecology and pathology of of ranaviruses in cell culture and tissue samples, J Virol Meth
amphibian ranaviruses, Dis Aquat Org 87:243–266, 2009. 171:225–233, 2011.
25. Duffus ALJ, Pauli BD, Wozney K, et al: Frog virus 3-like infec- 42. Allender MC, Abd-Eldaim M, Schumacher J: PCR prevalence of
tions in aquatic amphibian communities, J Wildl Dis 44:12, Ranavirus in free-ranging eastern box turtles (Terrapene carolina
2008. carolina) at rehabilitation centers in three southeastern US states,
26. Hausmann JC, Wack AN, Allender MC, et al: Experimental J Wildl Dis 47:759–764, 2011.
challenge study of FV3-like ranavirus infection in previously 43. Balseiro A, Dalton KP, del Cerro A, et al: Pathology, isolation
FV3-like ranavirus infected eastern box turtles (Terrapene caro- and molecular characterisation of a ranavirus from the common
lina carolina) to assess infection and survival, J Zoo Wildl Med midwife toad Alytes obstetricans on the Iberian Peninsula, Dis
46:732–746, 2015. Aquat Organ 84:95–104, 2009.
27. Cunningham AA, Hyatt AD, Russell P, et al: Emerging epi- 44. Cinkova K, Reschova S, Kulich P, et al: Evaluation of a polyclonal
demic diseases of frogs in Britain are dependent on the source antibody for the detection and identification of ranaviruses from
of ranavirus agent and the route of exposure, Epidemiol Infect freshwater fish and amphibians, Dis Aquat Organ 89:191–198,
135:1200–1212, 2007. 2010.
28. Cunningham AA, Langston TES, Bennett PM, et al: Pathological 45. Cunningham AA, Tems CA, Russell PH: Immunohistochemical
and microbiological findings from incidents of unusual mortality demonstration of Ranavirus Antigen in the tissues of infected
of the common frog (Rana temporaria), Phil Trans R Soc Lond frogs (Rana temporaria) with systemic haemorrhagic or cutaneous
Biol Sci 351:1539–1557, 1996. ulcerative disease, J Comp Pathol 138:3–11, 2008.
370 SE C T I O N 11 Amphibians and Reptiles
46. Hyatt AD, Williamson M, Coupar BEH, et al: First identifica- concurrent herpesvirus and Mycoplasma infection: management
tion of a ranavirus from green pythons (Chrondropython viridis), and monitoring, J Zoo Wildl Med 47:256–270, 2016.
J Wildl Dis 38:239–252, 2002. 58. Rojas S, Richards K, Jancovich JK, et al: Influence of temperature
47. Johnson AJ, Wendland L, Norton TM, et al: Development on ranavirus infection in larval salamanders (Ambystoma tigri-
and use of an indirect enzyme-linked immunosorbent assay for num), Dis Aquat Organ 63:95–100, 2005.
detection of iridovirus exposure in gopher tortoises (Gopherus 59. Bryan LK, Baldwin CA, Gray MJ, et al: Efficacy of select disin-
polyphemus) and eastern box turtles (Terrapene carolina carolina), fectants at inactivating Ranavirus, Dis Aquat Organ 84:89–94,
Vet Microbiol 142:160–167, 2010. 2009.
48. Whittington RJ, Speare R: Sensitive detection of serum antibod- 60. Fox SF, Greer AL, Torres-Cervantes R, et al: First case of
ies in the cane toad Bufo marinus, Dis Aquat Organ 26:59–65, ranavirus-associated morbidity and mortality in natural popula-
1996. tions of the South American frog (Atelognathus patagonicus), Dis
49. Whittington RJ, Kerns C, Speare R: Detection of antibodies Aquat Organ 72:87–92, 2006.
against iridoviruses in the serum of the amphibian Bufo marinus, 61. Greer AL, Collins JP: Habitat fragmentation as a result of biotic
J Virol Meth 68:105–108, 1997. and abiotic factors controls pathogen transmission throughout a
50. Allender MC, Mitchell MA: Hematologic response to experi- host population, J Anim Ecol 77:364–369, 2008.
mental infections of frog virus 3-like virus in red-eared sliders 62. Allender MC, Abd-Eldaim M, Kuhns A, et al: Absence of Rana-
(Trachemys scripta elegans), J Herp Med Surg 23:25–31, 2013. virus and herpesvirus in a survey of two aquatic turtle species in
51. Elion G: Acyclovir: discovery, mechanism of action and selectiv- Illinois, J Herp Med Surg 19:16–20, 2009.
ity, J Med Virol 1:2–6, 1993. 63. Schloegel LM, Daszak P, Cunningham AA, et al: Two amphibian
52. Beutner KR, Friedman DJ, Forszpaniak C, et al: Valaciclovir diseases, chytridiomycosis and ranaviral disease, are now globally
compared with acyclovir for improved therapy for herpes zoster notifiable to the World Organisation for Animal Health (OIE):
in immunocompetent adults, Antimicrob Agents Chemother an assessment, Dis Aquat Organ 92:101–108, 2010.
39:1546–1553, 1995. 64. Schloegel LM, Picco AM, Kilpatrick AM, et al: Magnitude of
53. Coupar BEH, Goldie SG, Hyatt AD, et al: Identification of a the US trade in amphibians and presence of Batrachochytrium
Bohle iridovirus thymidine kinase gene and demonstration of dendrobatidis and ranavirus infection in imported North Ameri-
activity using vaccinia virus, Arch Virol 150:1797–1812, 2005. can bullfrogs (Rana catesbeiana), Biol Conserv 142:1420–1426,
54. Tsai C-T, Ting J-W, Wu M-H, et al: Complete genome sequence 2009.
of the grouper iridovirus and comparison of genomic organiza- 65. Jancovich JK, Davidson EW, Parameswaran N, et al: Evidence
tion with those of other iridoviruses, J Virol 79:2010–2023, for emergence of an amphibian iridoviral disease because of
2005. human-enhanced spread, Molec Ecol 14:213–224, 2004.
55. Gaio C, Rossi T, Villa R, et al: Pharmacokinetics of acyclovir 66. Picco AM, Collins JP: Amphibian commerce as a likely source
after a single oral administration in marginated tortoises, Testudo of pathogen pollution, Conserv Biol 22:1582–1589, 2008.
marginata, J Herp Med Surg 17:8–12, 2007. 67. Picco AM, Karam AP, Collins JP: Pathogen host switching in
56. Allender MC, Mitchell MA, Yarborough J, et al: Pharmacokinet- commercial trade with management recommendations, Ecohealth
ics of a single oral dose of acyclovir and valcyclovir in North 7:252–256, 2010.
America box turtles (Terrapene sp), J Vet Pharmacol Therap 68. St. Amour V, Wong WM, Garner TWJ, et al: Anthropogenic
36:205–208, 2012. influence on prevalence of 2 amphibian pathogens, Emerg Infecti
57. Sim RR, Allender MC, Crawford LK, et al: Ranavirus epizootic Dis 14:1175–1176, 2008.
in captive eastern box turtles (Terrapene carolina carolina) with
53
Anuran Reproduction
ELLEN BRONSON AND CARRIE K. VANCE
T
here are more than 6700 known species of Anura which stages of the reproductive cycle might be compro-
(frogs and toads) in the world, of which at least mised if there is reproductive failure. This is not a trivial step
2000 are currently considered threatened or endan- because the environmental cues and breeding behaviors are
gered,1 more than any other vertebrate taxa. A number of often species specific, thus requiring a solid understanding
complex factors have led to amphibian declines around of the reproductive cycle for a given species.19
the world, including habitat loss, disease, climate change,
pesticide use, and pollution.2–5 As a result of these biotic Anuran Reproductive Physiology
and abiotic factors, many amphibian populations are now
extinct or have become critically endangered in their native With few exceptions, anurans (frogs and toads) exhibit
habitats.5 Because amphibians have developmental stages external fertilization, in which the female deposits an egg
that use both aquatic and terrestrial environments, they are mass (termed oviposition or spawning) while the male
often considered sentinels for ecosystem health. However, fertilizes it.10,19 Egg maturation, or oogenesis, is signaled
amphibians are also threatened by specific pathogenic by progesterone to resume meiotic division and is typically
microorganisms, including ranaviruses, the trematode triggered by a combination of factors such as photope-
Ribeiroia ondatrae, and the fungus Batrachochytrium dendro- riod and temperature. The egg grows in size due to the
batidis (Bd).6,7 Bd has spread across several continents at an accumulation of vitellogenin, produced by the liver in
alarming rate, causing extinctions of numerous amphibian response to estrogen. After the eggs are mature and the
species.8,9 As a hedge against further amphibian extinc- environmental conditions are right, the female will seek
tions, zoological parks, aquaria, and other institutions have out a male for breeding. A cascade of hormones through
established assurance colonies with the goal of maintaining the HPG axis leads to ovulation of the eggs from the ovary
viable populations in captivity for species sustainability into the oviduct, and the egg mass becomes surrounded
and future reintroductions.10,11 In some cases the captive by egg jelly.10,18,20 Spawning follows (release of eggs from
assurance colonies consist of a significant proportion, if not the cloaca), such that the eggs are laid as a loosely bound
all, of the remaining individuals of the rescued species.4,11 string. Typically the male amplexes the female to fertilize
To sustain these assurance colonies and maintain genetic the eggs, embracing the female tightly with the front legs
diversity for long-term species survival, successful reproduc- around the cranial coelom, and fertilization by the male
tion of founders and descendants is required. Despite the occurs as the eggs are expressed. Most temperate anurans
best efforts of the zoological community, many amphibian lay eggs in water; however, the various species display a
species fail to naturally reproduce in captivity.5 Conse- wide array of reproductive strategies. Most anurans leave the
quently, amphibian biologists, veterinarians, and research- fertilized egg mass following reproduction, although there
ers around the world have partnered to develop hormone are a few species that participate in protecting the embryos
therapies and assisted reproductive technologies (ARTs) until hatching of the larval (tadpole) stage.
for enhanced reproductive output and management of Elucidating the behavioral cues that each species is
genetic diversity.12–15 Amphibian reproduction is a complex responsive to can take many years and be very difficult to
sequence of events that couples environmental, social, and identify and replicate in the captive setting. Many species
physical cues leading to hormonal cascades associated with require specific levels or changes in humidity, temperature,
the hypothalamic-pituitary-gonadal (HPG) axis, eventually barometric pressure, water depth, and light length and
resulting in gamete development and release.14,16–18 Amphib- intensity.4,19,21 Other factors may include tactile or acoustic
ian ART spans a wide range of approaches (e.g., hormone characteristics such as male calling or spray and vibration
therapy, egg expression, in vitro fertilization), which are effects of a replicated waterfall. Temperate species often do
used to either facilitate or circumvent specific steps in the best after a period of low temperature inducing brumation
natural cycle when reproductive failure becomes apparent. or hibernation followed by slow warming to induce egg
An important aspect in implementing ART is identifying production, egg maturation, and breeding behaviors.22
371
372 SE C T I O N 11 Amphibians and Reptiles
Many tropical species breed at the onset of, or during, the cues to stimulate breeding, some amphibian species do
rainy season, so misting systems or rain chambers, as well not reproduce well in captivity without the use of external
as temperature changes, may replicate the natural environ- hormones. For example, the Puerto Rican crested toad (Pel-
ment and spur reproduction. tophryne lemur), Mississippi gopher frog (Lithobates sevosus),
Houston toad (Anaxyrus houstonensis), and Wyoming toad
Health Factors Affecting Reproduction have rarely bred in captivity without the use of exogenous
hormones.22,24,26 The hormone cascade involved in both
Poor health of amphibians may impair the normal hormone spermiation and ovipositioning is well-preserved among
cascade and result in cessation of oogenesis and spermato- species, although in the development of protocols, some
genesis. For example, an animal in poor health or poor species seem to have better success with one or the other
nutrition will likely abort the process of vitellogenesis or of the hormones that are commercially available. Human
egg maturation if fat stores are insufficient, and instead the chorionic gonadotropin (hCG) has LH-like activity on the
body will begin to resorb the eggs. Principal energy reserves ovary or testes. Because it is a mammal-derived protein,
must be redirected for survival and metabolism and will be much higher doses are required in amphibians to stimulate
directed away from nonessential activities, such as reproduc- a response compared with when the hormone is given
tion, which is energy demanding. Another important health to mammals (approximately 2000 times higher per kg).
aspect is egg retention in female anurans, which is similar The most commonly used hCG product is Sigma-Aldrich
to dystocia in other vertebrates. These retained eggs remain C-1063 (CAS Number 9002-61-3). LH and FSH specific
either attached to the ovary or are ovulated into the oviduct to amphibians have not been developed, and synthetic
but not deposited, and this state may lead to bacterial commercial forms of these products have proven to be inef-
infection and sepsis.23 Occasionally, reproductive hormones fectual in amphibians.23 The most frequently used hormone
are needed to stimulate the release of these eggs from the analog in amphibians is gonadotropin-releasing hormone
ovary or oviduct to preserve the health of the female.10,24 For agonist (GnRH-A), commonly and hereafter referred
example, there has been a clinical need for the administra- to as luteinizing hormone–releasing hormone analog
tion of exogenous hormones to release egg masses from (LHRHa), available as the commercial product Des-Gly
Panamanian golden frogs (Atelopus zeteki), Wyoming toads 10, D-Ala 6-LHRH ethylamide acetate hydrate (Sigma-
(Anaxyrus baxteri), boreal toads (Anaxyrus boreas), and Aldrich, L4513; CAS Number 79561-22-1). Purchased as
tomato frogs (Dyscophus spp.).4,10,23,25 Panamanian golden a lyophilized powder, it is reconstituted in sterile saline or
frogs tend to develop very large egg masses and have long phosphate-buffered saline prior to use.21 The reconstituted
periods of amplexus (weeks to months); thus the reproduc- liquid may be frozen but is stable only for 24 hours once
tive season is metabolically challenging for both sexes, and thawed. Another group of drugs that have been investigated
high mortality is seen due to decimation of fat supplies and more recently in anurans with promising but mixed results
decreased food intake. Frequently, females do not lay eggs are the dopamine antagonists combined with LHRHa.27,28
even when amplexed, so the energy demands may be even Dopamine inhibits GnRH synthesis and secretion from
higher for months while the eggs resorb.4 For these reasons the hypothalamus, thereby reducing or eliminating the
the administration of exogenous hormones has been used secretion of LH and FSH. By administering a dopamine
to stimulate egg release and decrease mortality. antagonist, the inhibition of GnRH is removed and the
effect of the exogenous hormone(s) can theoretically be
Hormonal Regulation of Reproduction enhanced.17,21,28,29 In anurans a combination of LHRHa and
the dopamine antagonist metoclopramide has been termed
Amphibians share many of the main reproductive neuropep- “Amphiplex” by Trudeau and has been tested in a number of
tides, pituitary hormones, and gonadal steroids with other amphibian studies.27–29 However, the efficacy of such com-
vertebrates. Gonadotropin-releasing hormone (GnRH) is binations is known to be variable in fish species, and some
produced in the neurons of the amphibian hypothalamus, mortality has been reported in amphibians, so future studies
which stimulates the release of luteinizing hormone (LH) will need to delineate the efficacy and safety in the various
and follicle-stimulating hormone (FSH) from the anterior anuran species. Administration of exogenous hormones is
pituitary gland (Fig. 53.1).18 LH and FSH have a stimulatory considered most effective if given intracoelomically with
effect on steroidogenesis in the gonads. In males, they lead a small-gauge needle (Fig. 53.2). Other administration
to the release of testosterone, causing spermatogenesis and routes, such as absorbance through the skin, injections into
sperm maturation and release, and they also play a role in the dorsal lymph sac, subcutaneously, or intramuscularly,
calling and amplexus.22 In females, these gonadotropins lead have been used but typically with lower success.21,30,31
to ovulation, egg transport through the oviduct, production
and secretion of egg jelly, and spawning (see Fig. 53.1).
Exogenous Hormone Induction of
Exogenous Hormone Use in Anurans Spermiation in Male Frogs
Despite the best efforts of curators, keepers, and veterinar- Following exogenous hormone administration, sperm are
ians to replicate the natural habitats and environmental released into the cloaca and are mixed with urine, forming
CHAPTER 53 Anuran Reproduction 373
Behavioral
Response Amphibian Brain
Gonadotropin-
releasing hormone GnRH Hypothalamus
(GnRH)
GnRH
Negative feedback loop
–T
–E
–E
– P4
– DTH
Gamete
+ FSH + LH
Release
Human chorionic
+E gonadotropin (hCG)
Liver
+ DTH
G
+E hC
Ovaries Testes
Vitellogenin Leydig +E+T
Sertoli +E
Ovaposition Spermiation
• Figure 53.1 Diagram of the hormone cascade through the hypothalamic-pituitary-gonadal (HPG) axis.
Gonadotropin-releasing hormone (GnRH) is released by the hypothalamus in response to environmental or
endogenous cues and binds receptors at the anterior pituitary. The gonadotropin hormones—luteinizing
hormone (LH) and follicle-stimulating hormone (FSH)—are released and act at the level of the gonads to
stimulate steroidogenesis in the testes, and follicular growth and ovulation in the ovaries and yolk formation
(vitellogenesis) in the liver. LH is the regulator of the Leydig cells, which produce testosterone (T) and
estrogens (E). Testosterone is transported to Sertoli cells and converted to dihydrotestosterone (DHT) and
estrogen which may regulate positive and negative feedback loops in anuran species.
spermic urine. Spontaneous expression of spermic urine males need sufficient recovery time (approximately 2 weeks)
occurs when the animal is picked up and held over a Petri between hormone injections or spermiation can temporarily
dish and is a result of the natural defense mechanism seen cease.21,35
upon handling, especially in toads. Spermic urine may also Select species-specific protocols for hormone-induced
be obtained by gentle palpation with light pressure to the spermiation are presented in Table 53.1. LHRHa is the
ventral coelom (Fig. 53.3). For frogs a small-gauge catheter hormone of choice for inducing natural mating in most
inserted into the cloaca and urinary bladder may be necessary zoos and aquaria because it elicits a stronger amplexus
to obtain urine.21,31 In most toad species, the highest con- response than does hCG.14,36 For instance, LHRHa initiates
centration of sperm can be found 3–9 hours after hormone a good amplexus response in American toads (Anaxyrus
administration, but in frog species the response is earlier, americanus) and Fowler toads (Anaxyrus fowleri) with lower
with sperm being released in the urine 30–90 minutes after sperm production, whereas hCG tends to generate better
hormone treatment.14,30,32–34 If hormones are used to induce sperm production but a lower behavioral response.13,34
sperm production for natural breeding or artificial fertil- Hence, hCG is excellent for collection of sperm for AF and
ization (AF), it is imperative to know this expected time cryopreservation. When hCG is combined with LHRHa,
frame to coordinate with the window of availability of eggs it may produce an even stronger effect than with LHRHa
from the female for a particular species.13,22,34 Moreover, alone.18,34 LHRHa is used for induction of spermiation and
374 SE C T I O N 11 Amphibians and Reptiles
TABLE
53.1 Hormones and Dosages Used to Induce Spermiation in Select Male Anuran Species
Exogenous
Scientific Name Species Hormone Dosage Reference
Anaxyrus americanus American toad hCG 300 IU Kouba 2012
Anaxyrus boreas Boreal toad LHRHa 300 IU Roth 2010
Anaxyrus fowleri Fowler’s toad hCG 300 IU Kouba 2009
Atelopus zeteki Panamanian golden frog LHRHa 4 µg/g Della Togna 2017
Atelopus zeteki Panamanian golden frog Amphiplex LHRHa 0.4 µg/g + MCP 10 µg/g Della Togna 2017
Atelopus zeteki Panamanian golden frog hCG 10 IU/g Della Togna 2017
Bufo baxteri Wyoming toad hCG 300 IU Browne 2006
Bufo baxteri Wyoming toad LHRHa 4 µg Obringer 2000
Ceratophrys cranwelli Chacoan horned frog LHRHa 0.1–0.5 mg/kg Waggener 1998
Ceratophrys ornata Argentina horned frog LHRHa 0.1–0.5 mg/kg Waggener 1998
Lepidobatrachus laevis Budgett’s frog LHRHa 0.375 mg (0.29–0.58 µg/kg) Waggener 1998
Lepidobatrachus llanensis Llanos frog LHRHa 0.6 mg/kg Waggener 1998
Lithobates pipiens Northern leopard frog hCG/LHRHa 500 IU LHRH + 10 µg LHRHa Kouba 2009
Lithobates pipiens Northern leopard frog LHRHa 0.1–0.4 µg/kg Waggener 1998
Lithobates pipiens Northern leopard frog Amphiplex LHRHa 0.4 µg/g + MCP 10 µg/g Trudeau 2010
Litoria raniformis Southern bell frog LHRHaLucrin 20 µg Mann 2010
Peltophryne lemur Puerto Rican crested toad hCG 4 IU/g SC Crawshaw 2008
Peltophryne lemur Puerto Rican crested toad LHRHa 0.1 µg/g SC Crawshaw 2008
Pseudophryne corroboree Corroboree frog hCG 20 IU/g in DLS Byrne 2010
Pseudophryne corroboree Corroboree frog LHRHa 5 µg/g in DLS Byrne 2010
Pseudophryne guentheri Günther’s toadlet LHRHa 2 µg/g Silla 2011
Pyxicephalus adspersus African bullfrog LHRHa 0.1–0.5 mg/kg Waggener 1998
Rana temporaria European common frog LHRHa 50 µg Shishova 2011
Xenopus laevis African clawed frog LHRHa 0.1–0.5 mg/kg Waggener 1998
Xenopus tropicalis Tropical clawed frog hCG 100 IU Browne 2007
male needs to be synchronized with the expected time of prior to the ovulatory dose if the cues that stimulate egg
egg laying, and the pair is typically encouraged to amplex recruitment are not available, such as in the off season or
naturally.21,34 When amplexus does not occur naturally, when hibernation is needed.10,19 For example, a single low-
the female can be administered hormone and allowed to dose priming injection of LHRHa followed by an ovulatory
either spontaneously oviposit in the absence of a male, or dose of LHRHa is sufficient to induce female Günther’s
the eggs can be expressed from the cloaca through gentle toadlets to spawn.37 Fowler’s toads typically receive two low-
pressure. Once collected, the eggs may be fertilized with dose priming injections of hCG 72 hours apart, followed
stored sperm by conducting AF. Spawning may be induced by a higher ovulatory dose of hCG + LHRHa 48 hours
using a single ovulatory dose of hCG or LHRHa, or it may later to trigger egg laying.21 Spawning without hibernation
require one or more lower concentration priming doses to has been achieved in female Wyoming toads using either
advance egg maturation prior to oviposition. Some species, one or two priming doses and an ovulatory dose of hCG
such as the American toad, Puerto Rican crested toad, or the and LHRHa.22,25 In Northern leopard frogs, both sexes
common coquí (Eleutherodactylus coqui), have been found were treated with LHRHa plus metoclopramide to induce
to reliably lay eggs after receiving only an ovulatory dose amplexus and egg laying within 48–96 hours.28 However,
of exogenous hormones.24,48,49 However, in other cases a LHRHa alone was sufficient to achieve the same outcomes
priming dose may be required approximately 24–96 hours in female Panamanian golden frogs.4
376 SE C T I O N 11 Amphibians and Reptiles
A B
C D
• Figure 53.4 Ultrasound images of various egg development stages in the Mississippi gopher frog
(Lithobates sevosus). Arrows indicate ovarian tissue at each grade of oocyte development. Fluid is
indicated by hypoechoic areas and tissue or eggs by hyperechoic areas. (A) Grade 0, no or minimal egg
development, ovarian tissue is uniform and undeveloped (arrow). (B) Grade 1, minimal egg development
as eggs begin to form in small pockets in the ovaries (arrow). (C) Grade 2, increasing egg development
where eggs begin to be distinguishable from tissue (arrow). (D) Grade 3, fully developed individual eggs
are seen as hyperechoic specks surrounded by fluid. Imaging was performed with a Sonosite MicroMaxx
ultrasound machine equipped with a 38-mm broadband linear array transducer (range 6–13 MHz). A scan
depth of 2.7 cm was used. (Image credit: Allison Julien, Mississippi State University.)
TABLE
53.2 Hormones and Dosages Used to Induce Oviposition in Select Female Anuran Species
Priming
Scientific Name Species Hormone Dose(s) Ovulatory Dose Reference
Anaxyrus americanus American toad hCG, hCG/LHRHa 100 IU (twice) 500 IU/20 µg Kouba 2009
Anaxyrus baxteri Wyoming toad hCG/LHRHa 500 IU/4 µg; 500 IU/4 µg Browne 2006
100 IU/
0.8 µg
Anaxyrus boreas Boreal toad LHRH 0.1 µg/g Roth 2010
Anaxyrus fowleri Fowler’s toad hCG, hCG/LHRHa 100 IU (twice) 500 IU/20 µg Kouba 2009
Anaxyrus fowleri Fowler’s toad hCG/LHRHa 500 IU/4 µg Browne 2006
Anaxyrus fowleri Fowler’s toad LHRHa 20 µg Browne 2006
Anaxyrus fowleri Fowler’s toad LHRHa/ 20–60 µg/5 mg Browne, Li 2006
Progesterone
Atelopus zeteki Panamanian LHRHa 4 µg 4 µg Bronson 2015
golden frog
Eleutherodactylus coqui Common coqui LHRHa 20 µg Michael 2004
Lithobates pipiens Northern leopard LHRHa/ 0.4 µg/g + Trudeau 2010
frog Metoclopramide 10 µg/g
Mixophyes fasciolatus Great barred frog hCG 100 IU 100 IU Clulow 2012
Peltophryne lemur Puerto Rican LHRHa 0.1 µg/g Crawshaw 2008
crested toad
Pseudophryne corroboree Corroboree frog LHRHa 1 µg/g 5 µg/g Byrne 2010
Pseudophryne guentheri Günther’s toadlet LHRHa 0.4 µg/g 2 µg/g Silla 2011
Xenopus laeves African clawed frog hCG 10 IU 100–200 IU Browne 2007
Xenopus tropicalis Tropical clawed hCG 200 IU Browne 2007
frog
directly onto the egg jelly and allowed to sit for 5 minutes protocols at the ready may both increase our success in
before flooding the dish with aged tap water. The cleavage maintaining and expanding healthy populations in captivity
rate of the fertilized embryos may be observed through a as well as saving genetic material for the future.
dissecting microscope 4–6 hours following insemination
of the eggs. AF has been performed successfully in several
amphibian species to date.10,32,33,40 Wyoming toad tadpoles References
produced from AF have been released into the wild.22 1. IUCN: IUCN red list of threatened species: Amphibians.
2016. Available at: www.iucnredlist.org/initiatives/amphibians/
analysis/red-list-status. (Accessed 15 April 2017).
Role of Zoo Veterinarians in 2. Daszak P, Berger L, Cunningham AA, et al: Emerging infectious
Anuran Reproduction diseases and amphibian population declines, Emerg Infect Dis
5:735–748, 1999.
Zoo and wildlife veterinarians play an important role in 3. Neveu A: Incidence of climate on common frog breeding:
both the development of protocols for the medical manage- long-term and short-term changes, Acta Oecologica 35:671–678,
ment of reproductive disorders, as well as in the support 2009.
4. Bronson E, Barrett K, Murphy K, et al: Reproductive manage-
of reproductive specialists in the development of ARTs.
ment of the Panamanian Golden Frog (Atelopus zeteki). In 2015
As amphibian species become increasingly imperiled in Proceedings Annual Conference AAZV, 2015, p 52.
our changing world and assurance colonies of endangered 5. Bishop PJ, Angulo A, Lewis JP, et al: The amphibian extinction
amphibians become one of our only hopes in maintaining crisis - what will it take to put the action into the Amphibian
these species and returning them to their native habitats in Conservation Action Plan?, SAPIENS 5:97–111, 2012.
the future, the further development of ARTs will continue 6. Schloegel LM, Daszak P, Cunningham AA, et al: Two amphibian
to increase in importance. Having appropriate reproductive diseases, chytridiomycosis and ranaviral disease, are now globally
378 SE C T I O N 11 Amphibians and Reptiles
notifiable to the World Organisation for Animal Health (OIE): Frog (Rana servosa) and sperm transfer for in vitro fertilization,
an assessment, Dis Aquat Organ 2010. Preprint. Reprod Fertil Dev 24:170, 2011.
7. Rosenblum EB, Voyles J, Poorten TJ, et al: The deadly chytrid 27. Trudeau VL, Schueler FW, Navarro-Martin L, et al: Efficient
fungus: a story of an emerging pathogen, PLoS Pathog 6:1, 2010. induction of spawning of northern leopard frogs (Lithobates
8. Voyles J, Young S, Berger L, et al: Pathogenesis of chytridio- pipiens) during and outside the natural breeding season, Reprod
mycosis, a cause of catastrophic amphibian declines, Science Biol Endocrinol 11:14, 2013.
326:582–585, 2009. 28. Trudeau VL, Somoza GM, Natale GS, et al: Hormonal induc-
9. Berger L, Speare R, Daszak P, et al: Chytridiomycosis causes tion of spawning in 4 species of frogs by coinjection with a
amphibian mortality associated with population declines in rain gonadotropin-releasing hormone agonist and a dopamine
forests of Australia and Central America, Proc Natl Acad Sci antagonist, Reprod Biol Endocrinol 8:1–9, 2010.
95:9031–9036, 1998. 29. Della Togna G, Trudeau V, Gratwicke B, et al: Effects of hor-
10. Kouba AJ, Vance CK, Willis EL: Artificial fertilization for monal stimulation on the concentration and quality of excreted
amphibian conservation: current knowledge and future consid- spermatozoa in the critically endangered Panamanian golden frog
erations, Theriogenology 71:214–227, 2009. (Atelopus zeteki), Theriogenology 91:27–35, 2017.
11. Grow S: Grow S, editor: Amphibian conservation, 2010, Associa- 30. Obringer AR, O’Brian JK, Saunders RL, et al: Characterization
tion of Zoos and Aquariums. of spermation response, LH release and sperm quality in the
12. Reid GM: Abstracts from the Amphibian Ark Biobanking American toad (Bufo americanus) and the endangered Wyoming
Advisory Committee Workshop Towards a Biobanking Strategy toad (Bufo baxteri), Reprod Fertil Dev 12:51–58, 2000.
for Amphibian Conservation, September 6–8, 2010 London and 31. Byrne PG, Silla AJ: Hormonal induction of gamete release
Portsmouth, UK, Biopreserv Biobank 10:62–69, 2012. and in-vitro fertilisation in the critically endangered southern
13. Kouba AJ, Lloyd RE, Houck ML, et al: Emerging trends for corroboree frog, Pseudophryne corroboree, Reprod Biol Endocrinol
biobanking amphibian genetic resources: the hope, reality 8:144, 2010.
and challenges for the next decade, Biol Conserv 164:10–21, 32. Shishova NR, Uteshev VK, Kaurova SA, et al: Cryopreservation
2013. of hormonally induced sperm for the conservation of threatened
14. Kouba AJ, Vance CK: Applied reproductive technologies and amphibians with Rana temporaria as a model research species,
genetic resource banking for amphibian conservation, Reprod Theriogeniology 75:220–232, 2011.
Fertil Dev 21:719–737, 2009. 33. Waggener WL, Carroll EJ: A method for hormonal induction
15. Holt WV, Brown JL, Comizzoli P, editors: Reproductive science in of sperm release in anurans (eight species) and in vitro fertiliza-
animal conservation, 2014. tion in Lepidobatrachus species, Dev Growth Differ 40:19–25,
16. Tsai P-S: Neuroendocrine control of reproduction in amphibians. 1998.
In Norris DO, Lopez KH, editors: Hormones and reproduction of 34. Kouba AJ, DelBarco-Trillo J, Vance CK, et al: A comparison of
vertebrates (vol 2). Amphibians. London, 2011, Elsevier Inc, pp human chorionic gonadotropin and luteinizing hormone releas-
21–37. ing hormone on the induction of spermiation and amplexus in
17. Vu M, Trudeau VL: Neuroendocrine control of spawning in the American toad (Anaxyrus americanus), Reprod Biol Endocrinol
amphibians in its practical applications, Gen Comp Endocrinol 10:59, 2012.
234:28–39, 2016. 35. McDonough CE, Martin MW, Vance CK, et al: Frequency
18. Langhorne C: Developing assisted reproductive technologies for of exogenous hormone therapy impacts spermiation in male
endangered North American anurans, 2016. Fowler’s toad (Bufo fowleri), Reprod Fertil Dev 28:995–1003,
19. Browne RK, Zippel K: Reproduction and larval rearing of 2016.
amphibians, ILAR J 48:214–234, 2007. 36. Mann RM, Hyne RV, Choung CB: Hormonal induction of
20. Rastogi RK, Pinelli C, Polese G, et al: Hormones and reproduc- spermiation, courting behavior and spawning in the southern
tive cycles in anuran amphibians. In Norris DO, Lopez KH, bell frog, Litoria raniformis, Zoo Biol 29:774–782, 2010.
editors: Hormones and reproduction of vertebrates (vol 2). Amphib- 37. Silla AJ: Effect of priming injections of luteinizing hormone-
ians. London, 2011, Elsevier, pp 171–185. releasing hormone on spermiation and ovulation in Gunther’s
21. Kouba AJ, Vance CK, Calatayud N, et al: Assisted reproductive Toadlet, Pseudophryne guentheri, Reprod Biol Endocrinol 9:68,
technologies (ART) for amphibians. In Amphibian Husbandry 2011. Available at: http://www.ncbi.nlm.nih.gov/pubmed/
Resource Guide, 2012, p 60. 21599916.
22. Browne RK, Seratt J, Vance CK, et al: Hormonal priming, 38. Kouba AJ, Vance CK, Frommeyer MA, et al: Structural and
induction of ovulation and in-vitro fertilization of the endan- functional aspects of Bufo americanus spermatozoa: effects of
gered Wyoming toad (Bufo baxteri), Reprod Biol Endocrinol 4:34, inactivation and reactivation, J Exp Zool A Comp Exp Biol
2006. 295:172–182, 2003.
23. Roth TL, Szymanski DC, Keyster ED: Effects of age, weight, 39. Germano JM, Arregui L, Kouba AJ: Effects of aeration and
hormones, and hibernation on breeding success in boreal toads antibiotics on short-term storage of Fowler’s toad (Bufo fowleri)
(Bufo boreas boreas), Theriogenology 73:501–511, 2010. sperm, Aquaculture 396–399:20–24, 2013.
24. Crawshaw G: Veterinary participation in the Peurto Rican 40. Sargent MG, Mohun TJ: Cryopreservation of sperm of Xenopus
Crested Toad program. In Zoo and wild animal medicine, current laevis and Xenopus tropicalis, Genesis 41:41–46, 2005.
therapy, St. Louis, MO, 2008, Elsevier, pp 126–136. 41. Shishova NR, Uteshev VK, Sirota NP, et al: The quality and
25. Browne RK, Li H, Seratt J, et al: Progesterone improves the fertility of sperm collected from European common frog (Rana
number and quality of hormone induced Fowler toad (Bufo temporaria) carcasses refrigerated for up to 7 days, Zoo Biol
fowleri) oocytes, Reprod Biol Endocrinol 4:3, 2006. 32:400–406, 2013.
26. Kouba AJ, Willis E, Vance CK, et al: Development of assisted 42. Mannion P: Diagnostic ultrasound in small animal practice,
reproduction technologies for the endangered Mississippi Gopher Oxford, 2001, Blackwell Science.
CHAPTER 53 Anuran Reproduction 379
43. Schildger B, Triet H: Ultrasonography in amphibians, Sem Avian analysis methods, Reprod Biol Endocrinol 1–14, 2016. Available
Exot Pet Med 10:169–173, 2001. at: http://dx.doi.org/10.1186/s12958-016-0174-9.
44. Stetter M: Diagnostic imaging of amphibians. In Amphibian 47. Marcec R: Development of Assisted Reproductive Technologies
medicine and captive husbandry, Malabar, FL, 2001, Krieger for Endangered North American Salamanders. 2016.
Publishing Company, pp 253–272. 48. Michael SF, Buckley C, Toro E, et al: Induced ovulation and
45. Reyer HU, Bättig I: Identification of reproductive status in female egg deposition in the direct developing anuran Eleutherodactylus
frogs – a quantitative comparison of nine methods, Herpetologica coqui, Reprod Biol Endocrinol 2:6, 2004.
60:349–357, 2004. 49. Johnson CJ, Kouba AJ, Vance CK, et al: Oviposition and ultra-
46. Graham KM, Kouba AJ, Langhorne CJ, et al: Biological sex sound monitoring of American toads (Bufo americanus) treated
identification in the endangered dusky gopher frog (Lithobates with exogenous hormones, Proc Am Assoc Zoo Vet 299–301,
servosa): a comparison of body size measurements, secondary 2002.
sex characteristics, ultrasound imaging, and urinary hormone
54
Minimally Invasive Surgery
of Amphibians
NORIN CHAI
M General Considerations
ore than 7645 species of amphibians exist of
which 6745 belong to the Anura (frogs and
toads), 695 to the Caudata (newts and salaman- In the author’s experience, the main indications of MIS
ders), and 205 to the Gymnophiona (caecilians).1 Only in amphibians are listed in Box 54.1. Depending on the
the commonly maintained companion animal species are equipment, the small size of the animal would be the most
discussed here; therefore caecilians have been excluded. The common contraindication. As usual, sick animals with high
term minimally invasive surgery (MIS) is generally used to anesthetic risks should not undergo laparoscopy. Despite
refer to any procedure that is less invasive than open surgery the variation in size and the nature of the procedures that
used for the same purpose. It should be safe and associated may be performed, the basic materials needed for MIS
with a lower postoperative patient morbidity compared are listed in Box 54.2. The amphibian patient should be
with a conventional approach for the same operation. In handled with care, and wrapping with wet paper towel is a
our case it will mostly refer to rigid endosurgical procedures, good technique to restrain an animal for a quick examina-
mainly laparoscopy. There have been sporadic reports of tion or for medication administration. Wearing moistened,
amphibian endoscopy since the 1980s. Most previous powder-free gloves prevents the transfer of microorgan-
reports describe the use of endoscopy to examine the gender isms or chemicals from the handler as well as protection
or retrieve foreign bodies.2–5 In contrast to reptile medicine, against secreted toxins. Manual restraint is used for short
where laparoscopy has been well developed, literature is still and nonpainful procedures. For a better visualization, it is
scarce in amphibians. The greatest limiting factor is prob- advisable to fast large frogs and toads for 24–48 hours prior
ably equipment compatibility due to the small size of most to anesthesia. General anesthesia with analgesia is required
amphibians. Still, with appropriate equipment, many species for MIS. Updated information on anesthesia, analgesia, and
of newts, salamanders, frogs, and toads may be internally basic and advanced surgical procedures has been recently
examined, with minimal trauma and discomfort. Laparos- described (see also Chapter 60).8 The benefits of MIS are
copy may be a useful complementary tool to radiography numerous, but most useful is the ability to collect samples
and ultrasonography. However, it is sometimes difficult for a definitive diagnosis, accurate prognosis, and to direct
to interpret images in such small animals. Misdiagnosis is therapy. In general, complications resulting from a properly
not uncommon. In the author’s experience, laparoscopy in performed laparoscopy procedure itself are rare. However,
amphibians permits visualization of almost all the coelomic iatrogenic endoscope trauma may adversely affect organs
organs from a single point of entry. In Fowler’s Zoo and and biopsy results, as well as result in hemorrhage. The most
Wild Animal Medicine, Volume 8, an overview of current dramatic complication occurs when the ventral abdominal
techniques and applications of MIS in wildlife has been vein is damaged. This may happen on very small animals,
provided, including a discussion on MIS-specific risks and even from a paramedian approach. The problem is that
disadvantages and on recent developments in human and there is no way to control the hemorrhage. If complica-
domestic animal surgery that have implications for wildlife tions occur, the procedure is immediately stopped. After a
surgery.6 The major concepts described may be appropriate warm, anesthetic-free bath or shower, the animal is soaked
here as well. Amphibian surgery is a very small special- in the balanced electrolyte solutions, the amphibian Ringer
ized field; MIS in amphibians is even smaller. Updated solution (6.6 g NaCl, 0.15 g CaCl2, 0.15 g KCl, and
information on endoscopy in amphibians has been recently 0.2 g NaHCO3 per liter of dechlorinated water), until the
described.7 This chapter will provide a brief overview of recovering of its biological functions.9 A major concern
current techniques of MIS in amphibians and an example is the risk-benefit ratio of proceeding with a MIS in an
of their application in fundamental research. amphibian. Cognitive bias has been previously discussed
380
CHAPTER 54 Minimally Invasive Surgery of Amphibians 381
incision site is localized by these two blue marks on this adult African
clawed frog (Xenopus laevis). (Copyright Norin Chai.)
• BOX 54.2 Standard Equipment
• 1.9-mm integrated telescope
• 2.7-mm diameter, 18-cm length, 30° oblique rigid telescope
with a 4.8-mm operating sheath
• Endovideo camera and monitor
• Xenon light source and light cable
• 1-mm or 1.7-mm endoscopic biopsy forceps and grasping
forceps. Endoscopic needle is optional.
• Carbon dioxide (CO2) insufflator with silicone tubing. Care
must be taken when using CO2 insufflation, because it may
quickly dry out the organs and the mucosa. In the author’s
experience, the endoscopic procedure should not last more
than 5 min. A simple syringe for air or saline infusion is also
practical.
recumbency, the left pelvic limb simply placed against its tail base. A
in wildlife MIS.6 This bias is also true in amphibians. The
line drawn between the shoulder and the hind limbs is divided into
author believes that amphibian MIS should be performed three equal parts, and the skin incision site is localized by the blue
by a practitioner already experienced at least with reptile mark. (Copyright Norin Chai.)
MIS. Practical recommendations should follow the edict
“first do no harm.” field is aseptically prepared by gently wiping the surgical site
with sterile gauze soaked in 0.75% chlorhexidine solution
Clinical Applications of Minimally Invasive and left on the surgical site for at least 10 minutes before
surgery. In dorsal recumbency, a 3-mm paramedian skin
Surgery in Amphibians incision is made in the mid-coelom (between the shoulders
Laparoscopic Examination and and the cloaca), either left or right (Fig. 54.1). Care must be
Endoscopic Biopsy taken not to damage the macroscopic glands, lymph hearts,
and blood vessels, especially the mid-ventral vein. On right
Laparoscopy may be defined as the exploration of the coe- lateral recumbency, the left pelvic limb is simply placed
lomic cavity using a rigid endoscope, indicated for many against the tail base. A line drawn between the shoulder and
diagnostic and surgical procedures. Despite the diversity in the hind limbs is divided into three equal parts (Fig. 54.2).
body shape, and the visceral anatomy and its distribution in The incision site is located on the boundary between the
amphibians, the process is similar for all species. The author second and third parts. In all cases the underlying muscle
uses dorsal recumbency for all anurans. In more laterally is grasped and elevated away from the coelomic viscera,
compressed amphibians (e.g., newts), lateral recumbency and small hemostats are gently forced through the coelomic
is used. By convention (or habit acquired with endoscopy musculature and into the coelomic cavity. The hemostats
in birds), the animal is placed on its right side. Insufflation are removed and replaced by the telescope within its sheath
is useful, but lower flow rates are used and maintained for and obturator (with insufflation line attached to one of the
intracoelomic pressures compared with those reported in ports) (Fig. 54.3). By making a small skin incision and
small animals. Increased intracoelomic pressure compresses breach in the muscle, the sheath will be tight-fitting and
the lungs because there is no true diaphragm. Further- insufflation gas leakage will be minimal. Typically, CO2
more, assisted ventilation is rarely possible during MIS in insufflation pressures of 0.5–2 mm Hg with a flow rate not
amphibians. With the animal positioned in dorsal or lateral exceeding 0.5 L/min are used. In some situations, saline
recumbency (depending on the body shape), the surgical infusion may be preferred over gas.
382 SE C T I O N 11 Amphibians and Reptiles
Amphibians have an undivided pleuroperitoneal cavity. The kidneys, paired, dark, flattened, with ovoid structures,
The only separated compartment is the pericardial sac. This are also located dorsocaudally (see Fig. 54.5F). The urinary
common coelom permits the visualization of liver, gallblad- bladder is large and thin-walled in anurans. Endoscopic
der, heart, lungs, digestive tract, gonads, kidneys, bladder, biopsy of parenchymatous organs or intracoelomic masses
and fat body from only a single entry point. may be performed. Hepatic or renal samplings submitted
In general, the liver of anurans consists of two completely for culture and histology are used for diagnosis antemortem
separated lobes (Fig. 54.4A–G).10 The liver of caudates, and postmortem. Collecting liver biopsy samples using
such as the axolotl (Ambystoma mexicanum), is a single laparoscopic surgery is technically easy to perform (see Fig.
elongated organ that may be partially subdivided. One must 54.4O). Laparoscopy provides adequately sized and lesion-
pay attention to the ventral vein at this point (see Fig. specific tissue samples for histopathologic analysis and other
54.4H and I). A large gallbladder lies on the midline in the tests (e.g., culture and heavy metal analysis).
interlobular connective tissue of the liver (see Fig. 54.4J). After examination, the scope is removed and the animal
It is important to look for the spleen (see Fig. 54.4K). deflates immediately. The coelomic membrane and the skin
The stomach is situated dorsally within the left side of the are closed in one layer with one or two interrupted sutures
coelom. The intestines fill the contralateral right side (see (Fig. 54.6). Monofilament nylon seems to be the most
Fig. 54.4L–N). The intestine may be subdivided into the appropriate suture in amphibian skin.11 Then the animal
narrow, coiled small intestine followed by a short, wide large is transferred to a warm, anesthetic-free bath and is rinsed
intestine that leads to the cloaca. The lungs lie dorsal to each copiously with fresh, well-oxygenated water.
lobe of the liver and the heart is further cranial, between the
shoulders. Each gonad (described later) is associated with Endoscopic Orchiectomy
a conspicuous fat body which is subdivided into numerous
digitiform lobes, pressed up against the pleuroperitoneal Endoscopic orchiectomy in a bullfrog (Lithobates catesbe-
cavity. The size of the fat bodies varies greatly with the stage ianus) is used here as an example of MIS. The animal is
of reproductive cycle. The ovaries vary in size, depending placed in dorsal recumbency. Prior to orchiectomy, a laparo-
on stage of the reproductive cycle, and may be massive, scopic examination is conducted and the gonads identified
occupying a large part of the pleuroperitoneal cavity. The as described previously (Fig. 54.7A). A second entry is
small, ovoid testes are less apparent, located in a dorsal achieved under direct endoscopic visualization and guided
position and thus covered by other viscera (Fig. 54.5A–E). by transillumination of the body wall, thereby avoiding
A B C
• Figure 54.3 (A) Laparoscopic examination of an African clawed frog (Xenopus laevis). A 3-lead system
electrocardiogram is used to assess the heart rate. The insufflation line is visible, attached to one of
the ports of the telescope’s sheath. (B) Laparoscopic examination of a Golden poison frog (Phyllobates
terribilis). Due to the size of the animal, the monitoring is difficult, thus minimal. (C) A small skin incision
allows the sheath to be tight-fitting. Please also note that the incision in this African clawed frog is more
cranial: it was for a biopsy of the myocardium. (Copyright Norin Chai.)
• Figure 54.4 (A) The liver of anurans consists of two completely separated lobes, here in an African
clawed frog (Xenopus laevis). (B and C) In the African clawed frog, the normal hepatic gross appearance
varies from pale gray, pink brown, to black. Note the normal dark pigmentation due to the presence of
melanomacrophages. This is common in most amphibians. (D) Hepatic cystic lesion in an African clawed
frog caused by Contracaecum sp. (E) Hepatic lipidosis in a wide-mouth frog (Lepidobatrachus laevis).
(F) Focal hepatic discoloration in an African clawed frog caused by Mycobacterium liflandii infection. (G)
Several hepatic abscesses in a western clawed frog (Xenopus tropicalis) caused by Mycobacterium
szulgaï infection. (H and I) The ventral vein travels cranially between the lobes of the liver to further divide
with a branch entering each lobe. (J) The large gallbladder often appears yellowish (normal). (K) Spleen
with multiple abscesses in an African clawed frog caused by Mycobacterium gordonae infection. (L) The
stomach (S) may be found caudal to the liver varying in size and shape depending on the species and
nature of the prey. Note the normal fat bodies (FB). (M and N) Coelomic organs with the small intestine
(SI), the wide large intestine (WI), and testis (T). (O) Biopsy of the liver. (Copyright Norin Chai.)
CHAPTER 54 Minimally Invasive Surgery of Amphibians 383
A B C
D E F
G H I
J K L
M N O
384 SE C T I O N 11 Amphibians and Reptiles
A B C
D E F
• Figure 54.5 (A) In mature females, the ova extend cranially, cover the parietal surface, and may
occupy large parts of the coelom as showed here in an African clawed frog (Xenopus laevis). (B) Gender
identification in immature great crested newt (Triturus cristatus). Note the immature ovaries that are
dorsally located. (C) Testis of an adult two-colored leaf frog (Phyllomedusa bicolor). (D) Ovarian abscess
in a western clawed frog (Xenopus tropicalis) caused by Mycobacterium szulgaï. (E) Ovarian neoplasia
in a two-colored leaf frog. (F) Kidney of a two-colored leaf frog. The kidneys are dark red cigar-shaped
cylinders located caudal, lateral to the spine. (Copyright Norin Chai.)
A B C
D E F
• Figure 54.7 Orchiectomy by minimal invasive surgery in a bullfrog (Lithobates catesbeianus). (A)
The abdominal cavity is evaluated and the testicles identified. (B) A second entry is achieved with a 11
scalpel blade under direct endoscopic visualization. (C and D) The gonad is held with atraumatic 5 mm
rigid grasping forceps. The surrounding blood vessels (E) are cauterized with a diode laser. (F) After
cauterization, the testicle is removed from the celom. (Copyright Norin Chai.)
main model organisms used in cardiac regenerative studies plane was indicated by the loss of withdrawal reflexes. For
are zebrafish (Danio rerio) and mice.13 Both show marked this procedure, the 3-mm paramedian skin incision was
differences, adult zebrafish having a highly efficient cardiac made just beneath the sternum. Care was taken to not
regenerative capacity, while adult mice lack this ability.12 damage the mid-ventral vein. Following skin incision, the
Amphibians are evolutionarily placed between these two abdominal membrane was elevated, incised, and carefully
organisms and have been historical models for heart devel- dissected. The telescope-sheath system was inserted into the
opment and cardiovascular physiology.14 Urodeles such as pleuroperitoneal cavity, which was insufflated with CO2.
the neotenic axolotl and adult eastern newt can regener- After endoscope insertion, the heart was located behind the
ate their heart ventricle.15 Although Xenopus sp. has been falciform ligament (Fig. 54.8A); then using the endoscopic
considered as a powerful model organism for regeneration biopsy forceps, the falciform ligament was opened and the
research over the past decades, only limited information is single ventricle chamber was immediately apparent (see Fig.
available on the outcome of cardiac injury in this species 54.8B and C). The pericardial sac was carefully pinched
and other frogs. The author has developed an endoscopy- open; then during the ventricle diastole, a biopsy was
based resection method to explore the consequences performed toward the heart apex (see Fig. 54.8D). Once
of cardiac injury in adult African clawed frog (Xenopus the scope was removed, the animal deflated immediately,
laevis). This method allowed in situ live heart observation, naturally removing the CO2. The coelomic membrane and
standardized tissue biopsy size, and reproducibility. The the skin were closed in one layer using interrupted sutures
procedures were performed with the same list of material with monofilament nylon. The animal was then transferred
described in Box 54.2. Frogs were fasted 24 hours prior to to an anesthetic-free bath and rinsed copiously with fresh,
anesthesia. Presurgical preparations included hydration of well-oxygenated dechlorinated water. The minimal invasive
the animal in a shallow dechlorinated water bath. Analgesia surgery itself (from incision to skin suture) took an average
was performed with an initial dose of butorphanol (1 mg/ of five minutes.
kg) followed by meloxicam (0.4 mg/kg), both injected into Finally, using this endoscopy-guided cardiac injury
the lymph sacs. After 10 minutes, anesthesia was performed procedure, it has been shown that the adult Xenopus sp.
by transferring the animals to a bath of buffered 1% tricaine heart was unable to regenerate.12 However, adult Xenopus
methanesulfonate (MS-222) for 8 minutes. Loss of righting sp. may complement adult mammalian models to study
reflex suggested a light stage of anesthesia. The surgical the consequences of heart injury after tissue removal.
386 SE C T I O N 11 Amphibians and Reptiles
A B C D
• Figure 54.8 Biopsy of the myocardium in an African Clawed Frog (Xenopus laevis). (A) Note the
heart behind the falciform ligament. (B and C) With an endoscopic biopsy forceps, the falciform ligament
is opened and the single ventricle chamber is immediately apparent. (D) The pericardial sac was carefully
pinched open; then during the ventricle diastole, a biopsy was performed toward the heart apex.
(Copyright Norin Chai.)
6. Romain P: Minimally invasive surgery techniques. In Fowler M, in the skin of the African clawed frog (Xenopus laevis), J Am Assoc
Miller R, editors: Zoo and wild animal medicine, ed 8, St. Louis, Lab Anim Sci 45(6):22–26, 2006.
2015, Saunders, pp 689–697. 12. Marshall L, Vivien C, Girardot F, et al: Persistent fibrosis, hyper-
7. Chai N: Endoscopy in amphibians, Vet Clin North Am Exot Anim trophy and sarcomere disorganisation after endoscopy-guided
Pract 18(3):479–491, 2015. heart resection in adult Xenopus, PLoS ONE 12(3):e0173418,
8. Chai N: Surgery in amphibians, Vet Clin North Am Exot Anim 2017.
Pract 19(1):77–95, 2016. 13. Gamba L, Harrison M, Lien CL: Cardiac regeneration in model
9. Wright KM: Overview of amphibian medicine. In Mader DR, organisms, Curr Treat Options Cardiovasc Med 16:288, 2014.
editor: Reptile medicine and surgery, ed 2, St. Louis, 2006, Saun- 14. Burggren WW, Warburton S: Amphibians as animal models for
ders Elsevier, pp 941–971. laboratory research in physiology, ILAR J 48:260–269, 2007.
10. Crawshaw GJ, Weinkle TK: Clinical and pathological aspects 15. Witman N, Murtuza B, Davis B, et al: Recapitulation of
of the amphibian liver, Semin in Avian and Exotic Pet Med developmental cardiogenesis governs the morphological and
9(3):165–173, 2000. functional regeneration of adult newt hearts following injury,
11. Tuttle AD, Law JM, Harms CA, et al: Evaluation of the gross Dev Biol 354:67–76, 2011.
and histologic reactions to five commonly used suture materials
55
Medical Aspects of the Hungarian
Meadow Viper Reintroduction
ENDRE SÓS AND BÁLINT HALPERN
Introduction and History of the Project animals further reduced the number of the vipers and
enhanced this negative trend. Current viper populations
The Hungarian meadow viper (Vipera ursinii rakosiensis; occur on diverse grasslands, where high prey abundance and
[HMV]) is a small and elusive venomous snake which was several different microclimates still provide an ideal habitat
first described by the famous Hungarian zoologist, Lajos for the species. Despite its name, areas with the lowest
Méhely in 1893.1 This taxon is considered a subspecies elevation and unpredictable water levels are dangerous for
within the intensively studied Vipera ursinii species complex HMVs and are therefore not preferred.
(Orsini viper), which includes lowland and mountain The HMV has been protected in Hungary since 1982,
members (Fig. 55.1). Aspects of the life history of the HMV strictly protected since 1986, and was raised to the highest
have only recently been understood (Table 55.1). conservation category in 1992. This viper is the most
Formerly, this species inhabited many suitable habitats endangered vertebrate species in the country and became
and its distribution covered areas in Romania and Austria directly threatened with extinction at the end of the 20th
as well. Until 2004 it was thought to be found only within century. Its critical situation was also recognized interna-
the boundaries of Hungary, but four small populations tionally; therefore it is included in the Bern Convention
were recently rediscovered in Transylvania, Romania. Appendix II,5 is listed in Appendix I of the Convention
Despite this finding, the current distribution area is much on International Trade in Endangered Species of Wild
more restricted than the historical range.2 There are only Flora and Fauna,10 and categorized as “Endangered” on
two remaining fragmented populations in the Hanság the International Union for Conservation of Nature and
and Kiskunság areas of Hungary, whereas in all the other Natural Resources (IUCN) Red List.9 The Bern Conven-
former habitats the species has become extinct. There are tion approved a European conservation action plan for the
two subpopulations found in the Hanság and nine in the meadow viper in 2005.8 In Hungary an appropriate species
Kiskunság areas. The total population size is estimated to conservation plan exists for the protection of this species,
be between 500 and 1000 individuals. As with many reptile and a complex conservation project was initiated, cofunded
taxa, an accurate census is difficult and population trends by European Commission’s LIFE and later LIFE+ funds.
are based on observations in the wild. Viper collectors from In April 1993, at the start of these complex conservation
the 1970s reported that 50 vipers could be found in 1 day efforts, the total HMV population was estimated at less
at some sites; however, in field studies at the end of the than 500 individuals. At that time, conservation experts
1990s, only one viper was found in 50 days spent searching. initiated a long-term conservation program and agreed
The main cause of population declines is the continuous on a number of principles, which included basic research,
loss of habitat due to the intensification of agriculture and learning about the life history of the species, and identifying
use of grasslands after World War II. Because this subspecies other possible threat factors and reasons for the severe and
is the inhabitant of steppe remnants, draining of marshy continuous population decline.
meadows and plowing of grasslands greatly reduced and
fragmented the suitable natural habitats, and mechanical Conservation Actions and Results
mowing techniques physically threatened the survival of
snakes and changed the structure of remaining habitats. The In 2001 a 2-day Population and Habitat Viability Assess-
illegal pet trade and the overall persecution of venomous ment (PHVA) meeting was organized at Budapest Zoo,
388
CHAPTER 55 Medical Aspects of the Hungarian Meadow Viper Reintroduction 389
350
300
250
Annual number of offspring
200
150
100
50
0
2004 2005 2006 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016
arising from small size of recent populations.7 Similar to treatment was also possible, but it was mainly restricted
many other species, juvenile mortality is the highest in the to captive or rescued animals at the HMVCC, or snakes
first year of life of the animals. Therefore young vipers born exhibited in two zoos (Budapest and Szeged) in Hungary.
at the HMVCC reached adulthood in higher percentage Because the conservation breeding program was lacking
than those in natural populations, due to prey abundance, even basic information, it was essential to collect micro-
lack of predators and separate keeping in terraria during biologic samples from seemingly healthy individuals from
their first winter. At the beginning of the program (between the beginning, when establishing a solid captive population
2004 and 2008), juveniles were housed indoors in a heated for breeding purposes. Each May between 2005 and 2008,
facility for their first winters and were released into outdoor, during the release process into seminatural terraria, cloacal
seminatural enclosures late in the following spring. This swabs, fecal samples, and skin impressions were collected to
practice helped to establish a relatively large and strong gather data about the health status of the young animals, as
population within a short time frame. After the rapid growth well as to obtain a clear picture of the normal enteral flora
of the captive population, this practice was neither possible and the parasite status of the snakes (these samples were used
logistically, nor important for the benefit of the program; for bacteriologic, parasitologic, and virologic studies). This
thus all vipers (including the youngsters) spent their first step was of utmost importance, even for the latter stages of
winter in artificial wintering burrows in their seminatural the project, because releases could pose a potential threat to
terraria. These circumstances closely mimic the natural native populations through the transmission of diseases. We
conditions but allow only limited control of the animals. found that cloacal swabs and fecal material were easily col-
An annual mortality of approximately 10% of juveniles was lected even from 9- to 10-month-old youngsters. The results
observed in the cohorts of 2004–2008, but since 2009 a did not yield the presence of pathogenic organisms, but
higher first-year mortality was recorded, although this level similarly to many other reptile species, Salmonella sp. was
in most of the years is considered still lower than in the wild, found to be part of the normal enteral flora of HMVs, and
where predation must also be factored in for this age group. all obtained samples were free of ophidian paramyxovirus
The breeding element of the conservation program was (OPMV). Regular parasitologic, bacteriologic, and virologic
a crucial contribution factor for the survival of the HMV. sampling of HMVs continues to be part of the medical
However, it had several medical implications, including the protocol of the HMVCC.
wild source populations, the captive conservation breeding
populations, and the intended animals for release. Screening, Diagnostics and Special Medical
quarantine, and other preventive medicine tools needed to Conditions
be implemented by the program overall (including incom-
ing founder animals, translocated individuals within the Due to the small size and anatomy of the HMV, clinical
HMVCC, and assigned individuals for release). Individual examination has its limitations. The primary site for blood
CHAPTER 55 Medical Aspects of the Hungarian Meadow Viper Reintroduction 391
• Figure 55.3 The delicate ventral aspect of the caudal vein is the • Figure 55.4 Ovariohysterectomy in the Hungarian meadow viper
preferred site for blood collection in the Hungarian meadow viper (Vipera ursinii rakosiensis).
(Vipera ursinii rakosiensis).
collection is the ventral aspect of the caudal vein (Fig. 55.3). dystocia in HMVs are not fully understood. However, based
Although blood can be drawn from the right palatine vein on our experience, although these females could theoretically
as well, it is not advisable, due to the close proximity of produce offspring in the subsequent season, the likelihood
the fangs and the collected blood is often mixed with snake of a repeated dystocia is very high; therefore we recommend
venom. Because only a small amount may be taken at a time ovariohysterectomy in these individuals (Fig. 55.4).
(maximum 1% of body weight), the samples were mainly Healthy newborn vipers usually complete their first
used for genetic analyses. Evaluation of the health status was shedding within 10 minutes of birth; however, we do
difficult because the collection of the proper volume may be encounter problems with shedding relatively frequently.
compromised; therefore sometimes the most relevant and The affected animals are emaciated, refuse to eat, lose weight
informative values were measured exclusively. Due to the rapidly, and are unable to go through the normal sloughing
small size of the species, blood collection from the heart is process. Usually 2–4 layers of dry, parchment-like skin may
contraindicated. be found on their bodies, which is strongly attached, and
Apart from the physical examination (including sam- it is impossible to simply remove it. Despite force feeding,
pling) and blood work, diagnostic imaging was generally topical hydration, warm baths, and other supportive treat-
very useful due to the small size of the species. Due to the ment, only approximately 30% of these affected animals
fact that the main goal of the program was conservation may be saved.
breeding, we used radiography and ultrasonography to During the late autumn of 2006, we lost 17 young
assess the reproductive tracts. We may conclude that, for vipers born that same year out of 39 individuals. The clini-
the detection of late pregnancy, both methods are suitable, cal symptoms were rather complex, including respiratory,
but because of the potential teratogen radiologic effect of integumentary, and neurologic signs. One key element
radiology, we strongly advise use of ultrasound equipment of the differential diagnosis was to exclude the possible
(with 8–11 MHz frequency probes). involvement of OPMV infection because it could have been
As a rare reproductive medical condition, dystocia was detrimental to the future success of the whole program.
observed several times and caesarean sections were per- Disease was detected only among the young individuals in
formed in these instances. The parturition process may take the inside part of the HMVCC, and spread was presumed
some time in vipers, and a relaxed environment is of utmost to be from neighboring individuals in terraria in one row.
importance. According to our protocol, dystocia is present The final conclusion of this massive mortality was a reptile
if labor is longer than 24 hours. The first such case took mite (Ophionyssus natricis) and secondary bacterial infection
place in August 2005, when parturition appeared to have (Fig. 55.5).
ended in an adult female after seven juveniles were born Apart from the problems with juveniles, in a few cases
and despite conservative medical treatment (oxytocin and mechanical trauma had to be treated in rescued animals.
calcium), surgery was needed to remove five more young, Injuries were caused by lawn mowers and attack by predators.
of which two were still alive. The surgical technique was For such cases, devising a safe anesthesia protocol was man-
performed similarly to other snake species by way of a datory. In our experience, for premedication, we recommend
paramedian laparotomy, visualization and elevation of the the combination of ketamine hydrochloride (20–40 mg/
uterus, removal of the fetuses, and suturing the wall of kg) and midazolam (1–2 mg/kg) be given intravenously or
the uterus and the coelomic wall. One must consider that into the coelomic cavity. Intravenous administration may
the proper closure of the wound edges is quite challenging greatly reduce induction times and dosage, but due to the
due to the thin muscle layer. The contributing factors of small size of the species, it is often not possible, and larger
392 SE C T I O N 11 Amphibians and Reptiles
visible under the scales. (Courtesy Nadia Robert.) Hungarian meadow viper (Vipera ursinii rakosiensis).
We strongly feel that the medical elements were very fauna species. Strasbourg, Bern Convention Standing Commit-
important bricks in the structure of the HMV project. A tee, Council of Europe. 1979, revised 2002.
proper medical program may ensure not only the health of 6. Kovacs T, Korsos Z, Rehak I, et al: Population and habitat
viability assessment for the Hungarian meadow viper (Vipera
the individuals of the conservation breeding program but
ursinii rakosiensis), Budapest 1–107, 2002.
provide a One Health concept for the whole population, 7. Ujvari B, Madsen T, Kotenko T, et al: Low genetic diversity
including released and genuinely wild individuals. threatens imminent extinction for the Hungarian meadow viper
(Vipera ursinii rakosiensis), Biol Conserv 105:127–130, 2002.
References 8. Edgar P, Bird B: Action plan for the conservation of the meadow
viper (Vipera ursinii) in Europe, 2005. Council of Europe,
1. Méhely L: A magyar fauna egy uj mergeskigyoja (Vipera rakosien- Strasbourg. Retrieved April 25, 2017, from https://wcd.coe.int/
sis M), Math Term Tud Ert 12(2/3):87–92, 1894. com.instranet.InstraServlet?command=com.instranet.CmdBlob
2. Korsos Z: Europa legveszelyeztetettebb mergeskigyoja a parlagi Get&InstranetImage=1305847&SecMode=1&DocId=1440882
vipera (Vipera ursinii rakosiensis), Term Ved Kozl 1(1):83–88, &Usage=2.
1991. 9. IUCN (International Union for Conservation of Nature): The
3. Ujvari B, Korsos Z: Use of radiotelemetry on snakes: A review, IUCN Red List of threatened species, 2016. Gland, Switzerland,
Acta Zool Academ Sci Hung 46(2):115–146, 2000. and Cambridge, UK. Retrieved April 25, 2017, from http://
4. Nilson G, Andren C: The Meadow and Steppe Vipers of Europe www.iucnredlist.org.
and Asia. The Vipera (Acridophaga) ursinii complex, Acta Zool 10. CITES (Convention on International Trade in Endangered
Academ Sci Hung 47(2–3):87–267, 2001. Species of Wild Flora and Fauna): Appendices I, II and III, 2017.
5. Council of Europe: Convention on the conservation of European Châtelaine, Geneva, Switzerland. Retrieved April 25, 2017, from
wildlife and natural habitats. Appendix II. Strictly protected https://cites.org/eng/app/appendices.php.
56
Ophidiomycosis
JEAN A. PARÉ
394
CHAPTER 56 Ophidiomycosis 395
snake [Thamnophis sauritus], water snakes [Nerodia spp.], Arthroconidia are hardy, persist in the environment, and
black rat snake, eastern fox snake, black racer [Coluber can survive freezing.
constrictor], northern pine snake [Pituophis m. melano- When, where, and how ophidiomycosis (SFD) appeared
leucus], mud snake [Farancia abacura], rainbow snake [F. in wild snakes in North America remains speculative. Mild
erytrogramma], ring-necked snake [Diadophis punctatus], skin lesions typical of hibernation sores/blisters in timber
eastern milk snake [Lampropeltis t. triangulum]), acrochor- rattlesnakes at emergence were confirmed as ophidiomyco-
dids (file snake, Acrochordus sp.), pythonids (ball python), sis, raising the possibility that this disease has been around
boids (green anaconda), and elapids (broad-headed snake). for some time and that climate change and other factors
Only recently has ophidiomycosis been diagnosed outside may have resulted in the increased expression and severity of
of North America, in wild European snakes, specifically infection. Spillover of infection from captive to free-ranging
grass snakes (Natrix natrix) in the British Islands and a dice snakes appears much less likely.
snake (N. tessalata) in the Czech Republic.9 It is likely that Wild snakes may become infected with Oo through
all snakes are susceptible.10 contact with sick snakes or with exuvia left behind by sick
snakes, with contaminated soil, or with fomites. Recent
Epidemiology use of newer, more sensitive molecular assays did disclose
the presence of Oo at very low levels on the skin of some
Available data indicate that infection of captive snakes with normal-looking snakes15,16; therefore the risk exists that
Oo antedates that of wild snakes by two decades or more.1,9 healthy carriers may introduce disease in a collection or
The two first known Oo isolates were collected a year apart, population. Koch’s postulates for Oo have been fulfilled in
in 1985 and 1986, from geographically distant sources: a experimental challenges of corn snakes and cottonmouths.6,7
captive ball python in the United Kingdom and a corn The severity of lesions in infected animals appears to vary
snake in western New York State. In Australia in 1976, between snake species, ranging from disfiguring in many
three wild-caught carpet pythons (Morelia spilotes), within a massasaugas to very mild in northern pine snakes; it also
few months in captivity, died of fungal dermatitis in which varies across individuals within the same species.
Oo-like arthroconidial tufts were histologically disclosed at
the skin surface.11 Geotrichum candidum, recovered from
these pythons, was likely a misidentification, as Oo had not Pathogenesis, Clinical Signs, and
yet been described. In fact, in the past, Oo and other former Disease Outcome
CANV-like fungi have been consistently mistaken in the lab
for other arthroconidiating fungi, such as Trichophyton spp., Conidia come in contact with intact or abraded/breached
Trichosporon spp., Geotrichum candidum, and Malbranchea snake skin; they germinate and colonize the epidermal
sp. A case of severe, deep granulomatous facial mycosis in corneum with proliferating vegetative hyphae that release
a captive western massasauga (Sistrurus c. tergeminus) in a keratinases and other proteases. This results in epidermal
Central Florida Zoo in 1979 was likely a case of ophid- necrosis and the recruitment of inflammatory cells, initially
iomycosis misdiagnosed as phycomycosis (zygomycosis).12 heterophils. Eventually hyphae move down into the deeper
Ophidiomyces ophiodiicola was likely present on several epidermal layers and then through to the dermis. Hyphal
continents as early as in the 1980s. The global trade in proliferation may extend to underlying muscles and bones,
pet reptiles might well have facilitated movement of this especially in massasaugas and other crotalids. Clinically,
pathogen. focal or multifocal and coalescing scale necrosis occurs. The
Oo is rarely recovered from the skin of healthy captive head and face seem particularly affected, maybe because the
snakes. It was isolated from only 1 of 91 healthy snakes head is the part of the body that first contacts a potential
sampled in one study, in contrast with aspergilli, penicillia, source of the fungus and is also most likely to present
and other ubiquitous saprophytes.13 This one isolate, from abrasions or other wounds. Scales and scutes are discolored,
an African rock python (Python sebae) in a southwestern swollen, with subsequent epidermal brown crusting (Fig.
zoo, remains the only Oo isolate not to have been cultured 56.1). Ulceration occurs, as sometimes does hyperkerato-
from an actual skin lesion.13 Attempts to identify Oo by sis with thickened crusts or eschars. Granulomas in the
means of polymerase chain reaction (PCR) from the skin subcutis may cause swelling, and infection may extend to
of 31 wild massasaugas that were all free of lesions were underlying muscles and bone and result in disfiguration.
unsuccessful,14 further underlining a strong association Very rarely does terminal fungemia or dissemination of the
between the presence of this fungus on the skin of snakes fungus to internal viscera occur. Snakes typically remain
and the presence of lesions/disease. Many case reports of alert until disease has progressed. Lesions on the head and
ophidiomycosis in captive snakes involve recently captured often around the nares and the nasolabial pits in crotalids
animals, suggesting that the fungus was carried by these progressively impede hunting and feeding, so that animals
animals and that the stress of captivity may have resulted lose condition and die.
in disease. Ophidiomycosis is contagious and can spread Infected snakes often undergo more frequent ecdysis, as
among captive snakes. Propagation is likely by means if to better rid themselves of skin fungal loads. Infection also
of arthroconidia on the surface of infected or shed skin. leads to behavioral changes and aberrations that may affect
396 SE C T I O N 11 Amphibians and Reptiles
B
• Figure 56.3 (A) Right lateral view of the head of a free-ranging corn
snake, Pantherophis guttatus, with ophidiomycosis. (B) Right lateral
view of the head of the same corn snake, P. guttatus, postecdysis. The
hyphema resolved and the snake was released. ([A] Photo courtesy of
Michael Bisignano; [B] Photo courtesy of Julie Larsen Maher, WCS.)
presence of arthroconidia at the surfaces of lesions, whether relationship with some human-associated isolates, J Clin Micro-
on cytology of impression smears from lesions or in tissue biol 51:3338–3357, 2013.
sections, is highly suggestive of ophidiomycosis. A firm 2. Rajeev S, Sutton DA, Wickes BL, et al: Isolation and charac-
terization of a new fungal species, Chrysosporium ophiodiicola,
diagnosis relies on culture or PCR from clinical or necropsy
from a mycotic granuloma of a black rat snake (Elaphe obsoleta
material and demonstration of morphologically compat- obsoleta), J Clin Microbiol 47:1264–1268, 2009.
ible fungal elements in tissue sections. Conventional PCR 3. Allender MC, Dreslik M, Wylie S, et al: Chrysosporium sp. infec-
assays are available, and newer and more sensitive qPCR tion in massasauga rattlesnakes, Emerg Infect Dis 17:2383–2384,
and TaqMan PCR assays have recently been developed.15,16 2011.
Ideally, in situ hybridization for Oo in tissue sections would 4. McBride MP, Wojick KB, Georhoff TA, et al: Ophidiomyces
best demonstrate causation. ophiodiicola dermatitis in eight free-ranging timber rattlesnakes
(Crotalus horridus) from Massachusetts, J Zoo Wildl Med
46:86–94, 2015.
Treatment 5. Sleeman J: Snake Fungal Disease in the United States. National
Wild snakes with mild lesions at emergence may improve Wildlife Heath Center Wildlife Health Bulletin, 2013-02,
3 pages.
through successive shedding cycles over the summer and
6. Lorch JM, Lankton J, Werner K, et al: Experimental infection
may therefore not require treatment. Hospitalization may, of snakes with Ophidiomyces ophiodiicola causes pathological
however, be required for snakes with more severe lesions changes that typify snake fungal disease, MBio 6(6):e01534–15,
and captive snakes with ophidiomycosis. Strict isolation and 2015, doi:10.1128/mBio.01534-15.
proper biosecurity measures should be instituted to curtail 7. Allender MC, Baker S, Wylie D, et al: Development of Snake
the risk of contagion. A number of disinfectants—such as Fungal disease after experimental challenge with Ophidiomyces
bleach, ethanol, quaternary ammonium, and others—have ophiodiicola in cottonmouths (Agkistrodon piscivorus), PLoS ONE
demonstrated activity against Oo.17 The same compounds 10(10):e0140193, 2015, doi:10.1371/journal.pone.0140193.
should be used in the field to disinfect boots and gear 8. Allender MC, Raudabaugh DB, Gleason, et al: The natural
between sites. history, ecology, and epidemiology of Ophidiomyces ophiodiicola
Treatment consists of general supportive measures along and its potential impact on free-ranging snake populations,
Fungal Ecol 17:187–196, 2015.
with systemic and topical antifungals. Sick snakes should
9. Paré JA, Sigler L: An overview of reptile fungal pathogens in
be provided with fluid, thermal, and nutritional support the genera Nannizziopsis, Paranannizziopsis, and Ophidiomyces, J
as needed. This may be enough to achieve improvement Herpetol Med Surg 26:46–53, 2016.
with each ecdysis cycle when lesions are mild. Surgical 10. Franklinos LHV, Lorch JM, Bohuski E, et al: Emerging fungal
debridement of lesions is indicated when shedding fails to pathogen Ophidiomyces ophiodiicola in wild European snakes,
yield improvement, to remove crusts and eschars in which Sci Rep 7:3844, 2017, doi:10.1038/s41598-017-03352-1.
conidia would otherwise remain sequestered. Topical anti- 11. McKenzie RA, Green PE, Branch P: Mycotic dermatitis in captive
septics may be useful on wounds following debridement. carpet snakes (Morelia spilotes variegata), J Wildl Dis 12:405–408,
1976.
Itraconazole, voriconazole, and terbinafine were all active
12. Williams LW, Jacobson E, Gelatt KN, et al: Phycomycosis in a
in vitro against Oo isolates,18 but delivery of these oral western massasauga rattlesnake (Sistrurus catenatus) with infec-
compounds is problematic in venomous snakes. Osmotic tion of the telencephalon, orbit, and facial structures, Vet Med
pumps and implants for parenteral delivery of voricon- Small Anim Clin 74:1181–1184, 1979.
azole and terbinafine, respectively, are being investigated. 13. Paré JA, Sigler L, Rypien KL, et al: Cutaneous mycobiota of
Broad-spectrum antibiotics may help to prevent or treat captive squamate reptiles with notes on the scarcity of Chryso-
potential secondary bacterial infections. Duration of treat- sporium anamorph of Nannizziopsis vriesii, J Herpetol Med Surg
ment should extend beyond resolution of skin lesions and 13:10–15, 2003.
may require overwintering sick wild snakes in a captive 14. Allender MC, Dreslik MJ, Wylie DB, et al: Ongoing health
environment. Snakes with lesions should not be allowed, assessment and prevalence of Chrysosporium in the Eastern mas-
whenever possible, to hibernate. sasauga (Sistrurus catenatus catenatus), Copeia 101:97–102, 2013.
15. Allender MC, Bunick D, Dzhaman E, et al: Development and
use of a real-time polymerase chain reaction assay for the detec-
Conclusion tion of Ophidiomyces ophiodiicola in snakes, J Vet Diagn Invest
27:217–220, 2015.
Ophidiomycosis is a progressive and contagious dermato- 16. Bohuski E, Lorch JM, Griffin KM, et al: TaqMan real-time poly-
mycosis of captive and free-ranging snakes for which the merase chain reaction for detection of Ophidiomyces ophiodiicola,
outcome is often fatal. This disease should figure promi- the fungus associated with snake fungal disease, BMC Vet Res
nently on the list of differential diagnoses for snakes with 11:95, 2015, doi:10.1186/s12917-015-0407-8.
skin lesions. 17. Rzadkowska M, Allender MC, O’Dell M, et al: Evaluation of
common disinfectants effective against Ophidiomyces ophiodi-
References icola, the causative agent of snake fungal disease, J Wildl Dis
52:759–762, 2016.
1. Sigler L, Hambleton S, Paré JA: Molecular characterization 18. Paré JA, Andes DR, Sigler L: In vitro susceptibility of fungal
of reptile pathogens currently known as members of the isolates from reptiles to antifungal drugs, Proc AAZV, AAWV,
Chrysosporium anamorph of Nannizziopsis vriesii complex and AZA/NAG 124, 2005.
57
Fibropapillomatosis in Marine Turtles
ANNIE PAGE-KARJIAN
F
ibropapillomatosis (FP) in marine turtles is a debili- using molecular technologies such as polymerase chain reac-
tating, infectious disease characterized by single or tion (PCR) and in situ hybridization.25,26 The virus has been
multiple tumors that may develop anywhere on an visualized in tumors via transmission electron microscopy
afflicted turtle’s body (Fig. 57.1). FP mainly affects green (TEM), and immunohistochemistry and reverse transcrip-
turtles (Chelonia mydas) but has been reported in all tase PCR have further demonstrated that ChHV5 is tran-
marine turtle species.1–7 There is evidence that FP does not scriptionally active in epithelial cells of FP tumors.21,26–31
negatively impact green turtle population recovery, survival FP is a complex disease wherein multiple factors likely play
probability, or somatic growth; however, FP disease may a role in tumor development and progression, including
have severe negative effects on the health of an individually ChHV5 infection as well as environmental, microbial, and/
afflicted turtle.8–11 FP tumors are histologically benign; or immune-related cofactors (see also Chapter 39).
however—depending on their location, size, and degree
of invasiveness—they can be fatal in some cases.1 For Epidemiology
example, turtles with periocular or corneal tumors (Fig.
57.2) may have difficulty acquiring food and/or avoiding FP disease occurs worldwide but is mainly reported in
boats; turtles with oropharyngeal masses may have difficulty warmer waters in and around the tropics.1,32 The prevalence
feeding and/or breathing; turtles with flipper tumors may of FP disease has reached epizootic proportions in several
have reduced swimming ability and/or be more likely to green turtle populations.1,33 FP seems to be primarily a
become entangled in fishing gear; and turtles with internal disease of juvenile green turtles following their migration
tumors may experience organ dysfunction and/or physi- to near-shore habitats.34 Although FP was once identified
ologic imbalances.12,13 In free-ranging marine turtles, FP as a major cause of green turtle strandings in Hawaii, more
is most frequently observed in juveniles, and the presence recent studies show that prevalence of the disease in Hawai-
of FP-afflicted turtles has been associated with shallow/ ian green turtles is now in decline.8,35 Two plausible explana-
inshore waters, especially habitats affected by anthropogenic tions for this include the development of herd immunity and
impacts such as agricultural, urban, and industrial develop- the removal of a tumor-promoting environmental insult in
ment.1,14–17 FP is a major concern for caretakers of captive or the near-shore turtle foraging habitats.8,36,37 FP prevalence
rehabilitating turtles because extensive quarantine measures seems to be more stable in green turtles in the southeastern
are necessary for marine turtles with FP, and prognoses for United States, with very high prevalence in some areas of
turtles with severe FP are complicated by poor nutritional Florida, where FP is considered a major cause of mortality.11
condition, poor general health on admission, and secondary In other locations, FP prevalence is increasing, and it has
or opportunistic infections.18–21 recently been reported from numerous new localities in the
Atlantic and Pacific Oceans.16,38–40 In many studies, ChHV5
Etiology DNA has been detected via molecular techniques in tissue
samples collected from marine turtles with FP and from
Most evidence points to a herpesviral etiology for FP, with turtle populations in which FP prevalence is high.29,41–43
the majority of molecular data suggesting that chelonid ChHV5 prevalence independent of FP tumor prevalence
herpesvirus 5 (ChHV5) is the main causative agent of FP. has also been reported; for example, ChHV5 has been
A series of transmission experiments demonstrated three detected in normal skin biopsies from nontumored turtles
of the four Koch’s postulates, and in vitro replication of in populations where FP is common.43,44
the virus was demonstrated when de novo formation of FP tumors are a proven source of infectious ChHV5
ChHV5-positive intranuclear inclusions were observed in particles, and direct, horizontal transmission of ChHV5 was
three-dimensional cultures of green turtle skin cells.1,21–23 demonstrated in a series of infectivity trials.20–22,28,36,45 The
A consistently strong statistical association of ChHV5 with high prevalence of active ChHV5 infection in early tumors
FP tumors has been confirmed by many subsequent studies suggests that virus production and shedding predominantly
398
CHAPTER 57 Fibropapillomatosis in Marine Turtles 399
Clinical Signs
FP tumors are typically proliferative masses that can occur
anywhere on or within the turtle’s body (see Fig. 57.1).1,45,48
• Figure 57.2 Fibropapilloma lesions on the left eye of a juvenile green
Morphologically, FP tumors may have a wide variety of
turtle (Chelonia mydas) from the Florida Keys. Multilobulated verrucous gross appearances: flat plaques (Fig. 57.3), pedunculated,
tumors cover the epithelial surfaces of the conjunctiva, palpebrae, sessile, verrucous, smooth, or polypoid nodules, or a
sclera, and cornea, partially obscuring the turtle’s vision. Cutaneous combination of multiple types. The number, color, and
tumors may also be seen on the left and right front flippers. (Photo
size of FP masses may vary widely, depending on tumor
credit The Turtle Hospital.)
location and severity of disease. Secondary invaders such as
bacteria and/or fungi readily infect ulcerated FP lesions.27
occur early in the progression of FP disease.26,28 It has also The typical histologic description of cutaneous FP tumors
been postulated that ChHV5 transmission within a popula- includes papillary epidermal hyperplasia supported by
tion may largely depend on a few highly infectious indi- broad fibrovascular stalks with a varying ratio of epidermal
viduals with small tumors (<20 cm2 surface area) permissive to dermal proliferation.1,28 Lymphocytes and macrophages
to viral production.26 In areas where FP is endemic, viral may be found at tumor margins and infiltrating tumors in
shedding into the surrounding environment via sloughing moderate to marked numbers. Histologic evidence of clini-
of virally infected epidermal cells from FP tumors represents cal regression is observed in some tumors.19 Visceral tumors
a key source of infection; in areas with high FP prevalence, are perceived as more chronic lesions that develop following
epizootic transmission cycles are likely perpetuated in this cutaneous tumor proliferation.1,13,28 Histologic descriptions
way.34 ChHV5 transmission may be magnified by mechani- of visceral tumors include fibromas, myxofibromas, and
cal vectors such as marine leeches (Ozobranchus spp.).30 fibrosarcomas.1,18,28,49
Tissues other than cutaneous tumors may also be involved Stranded and free-ranging sea turtles with FP are often
in ChHV5 replication and transmission cycles, as suggested debilitated and/or cachectic. Severe FP has been associated
400 SE C T I O N 11 Amphibians and Reptiles
with various abnormalities in clinical pathology data, course of FP disease and host immunosuppression, stress,
including anemia, leukopenia, lymphopenia, eosinopenia, and comorbid conditions can lead to viral reactivation.58
and heterophilia.32,48,50 Hypoproteinemia, hypocalcemia, Supportive care of turtles with FP should include water
hypoalbuminemia, and hyperglobulinemia may also be of suitable quality and temperature, adequate and species-
observed in green turtles with FP.32,48–53 Suggestive of appropriate nutrition, fluid therapy, pain management, and
anemia of chronic disease and antigenic stimulation, these treatment of secondary infections.59 Antiviral therapeutics
changes are compatible with the clinical presentation of (e.g., L-lysine, acyclovir) may be used to supplement sup-
FP. Turtles afflicted with severe FP often present with a portive care; however, to date no controlled studies have
number of associated comorbidities, including bacterial, been performed on the efficacy of these treatments against
fungal, and/or parasitic coinfections, ileus, buoyancy issues, FP lesions.19
and boat-strike trauma.19 Although evidence suggests that some marine turtles with
less severe FP will undergo spontaneous FP lesion regression,
Diagnosis this should not be expected in most cases.13,19,60,61 Surgical
excision is currently the most effective way to treat cutane-
Although cutaneous FP may easily be recognized on gross ous, oral, and ocular FP lesions. Local or general anesthesia
examination, definitive diagnosis requires histopathol- can be used, depending on the size, number, and degree of
ogy findings compatible with FP. Follow-up diagnosis of invasiveness of the tumors. Multiple tumor excisions tend to
ChHV5 DNA using molecular techniques is also recom- require general anesthesia. The technique of choice is carbon
mended. If possible, all rehabilitating turtles with FP should dioxide (CO2) laser–mediated tumor removal; other options
be imaged to rule out visceral tumors. Widely available include sharp excision, cryosurgery, radioscalpel, electroche-
imaging techniques include radiography and ultrasonogra- motherapy, and electrocautery.13,19,62 The CO2 laser helps
phy; however, small soft tissue masses may evade diagnosis minimize hemorrhage to the tumor removal site as it simul-
with these techniques. Observation of suspicious inter- taneously cauterizes and seals the excision site or sites while
nal lesions on imaging may be followed by laparoscopy it cuts tissue.13 Laser power, pulse rate, and handpiece size
and biopsy.19 Endoscopic examination also has limitations, may be varied according to surface area extent and depth
however: dorsal lung and extraparenchymal lesions may be of the tumor or tumors. Plaque-like or broad-based FP
missed, and endoscopic procedures are not recommended lesions may be ablated using a lower power, whereas pedun-
in severely debilitated turtles.13,19 If available, computed culated or narrow-based tumors may be excised using a
tomography (CT) or magnetic resonance imaging (MRI) higher-power technique. Extreme care should be exercised
may be preferred, as these techniques do not require anes- in removing ocular lesions, including low power and low
thesia and tend to be more accurate in identifying small pulse rate, avoiding corneal tissue by ablating ocular tumors
internal tumors.54,55 at an angle. Sutures are usually not needed, and tumor exci-
ChHV5 infection may be inferred via identification of sion sites may be left open to heal by secondary intention;
viral DNA in biological swabs using molecular diagnos- however, sutures may be required in the case of removal of
tic techniques. For example, ChHV5 DNA has been dem- a very deep tumor. Perioperative analgesics and antibiot-
onstrated in cloacal, oral, and ocular swabs taken from ics should be administered. Careful postoperative monitor-
turtles with cutaneous FP lesions, although these sample ing should be implemented after tumor removal, including
types are not as sensitive for ChHV5 DNA as tumor and/ dry-docking patients for up to 24-hours postsurgery. Cuta-
or skin biopsies.25,56 Suspected cases of ChHV5 infection neous lesions may heal completely in as little as 12 weeks.
must be confirmed via direct visualization of areas of her- It is acceptable to perform multiple tumor removal surger-
pesvirus morphogenesis using histopathology and/or TEM ies, and this may be preferred in turtles with large tumor
in combination with molecular diagnostics such as PCR, burdens.13 The number of tumor removal surgeries was not
in situ hybridization, or immunohistochemistry. There are significantly related to prognosis in one study.19 A 4- to
several validated and published PCR assays targeting dif- 6-week period of healing should be allowed between surger-
ferent ChHV5 genes, although assays offered by commer- ies.13 An important caveat of tumor removal surgery is the
cial diagnostic laboratories may be less specific than those possibility of tumor regrowth: one study found that 38.5%
used in research.14,25,29,30,34,41,42,57 Confirmatory sequenc- of green turtles that underwent tumor removal surgery expe-
ing of PCR amplicons is required for proper diagnosis rienced FP regrowth within an average of 36 days postsur-
of ChHV5 infection. A serologic immunoassay based on gery.19 Although regrown tumors can be surgically removed,
recombinant antigen has been validated for detecting anti- it is best to avoid repeated cycles of tumor removal and
bodies to ChHV5 glycoprotein H but is not currently com- regrowth. To help prevent tumor regrowth, a wide margin of
mercially available.46 apparently normal tissue should be included in tumor exci-
sion whenever possible, as normal skin surrounding tumors
Treatment may serve as a source of ChHV5-infected cells.41 The like-
lihood of tumor regrowth may also be reduced by lowering
Supportive care is essential for the successful treatment of tank water temperatures by 2°C–5°C after tumor removal
FP, as the overall health of the turtle may strongly affect the surgery to help prevent viral reactivation.19
CHAPTER 57 Fibropapillomatosis in Marine Turtles 401
Prognostic Indicators and 2. Aguirre AA, Spraker TR, Chaves A, et al: Pathology of fibropapil-
lomatosis in olive ridley turtles (Lepidochelys olivacea) nesting in
Release Considerations Costa Rica, J Aquat Anim Health 11:283–289, 1999.
3. Barragan AR, Sarti ML: A possible case of fibropapilloma in
Tumor number, anatomic distribution, morphologic appear-
Kemp’s ridley turtle (Lepidochelys kempii), Mar Turtle Newsl
ance, and progression, as well as overall body condition and 67:27, 1994.
severity of comorbid conditions should dictate triage cri- 4. D’Amato AF, Moraes-Neto M: First documentation of fibropap-
teria for FP-afflicted marine turtles. If left untreated (and illomas verified by histopathology in Eretmochelys imbricata, Mar
in some cases regardless of treatment), certain tumor loca- Turtle Newsl 89:12–13, 2000.
tions and morphologies are associated with poor case out- 5. Harshbarger JC: Sea turtle fibropapilloma cases in the registry of
comes; these include visceral or intraocular tumors, tumors tumors in lower animals, in Proceedings, Research Plan for Marine
that have eroded thorough bony structures (e.g., carapace, Turtle Fibropapilloma. NOAA Tech Memo, 1991, 63–70.
plastron), and aggressive recurrent tumors.13,19 Turtles with 6. Huerta P, Pineda H, Aguirre A, et al: First confirmed case of
these lesions may be considered outright euthanasia can- fibropapilloma in a leatherback turtle (Dermochelys coriacea), in
didates. Other tumor types are more easily treated and Proceedings, 20th Annual Symposium on Sea Turtle Biology and
Conservation, NOAA Tech Memo, 2002, p 193.
should be considered on a case-by-case basis, taking into
7. Limpus CJ, Couper PJ, Couper KLD: Crab Island revisited:
account comorbid conditions, available resources and treat- reassessment of the world’s largest flatback turtle rookery after
ment options, and quarantine capabilities. Although in one twelve years, Mem Queensl Mus 33:227–289, 1993.
study, green turtles with ocular tumors were significantly 8. Chaloupka M, Balazs GH, Work TM: Rise and fall over 26 years
less likely to survive rehabilitation than turtles with tumors of a marine epizootic in Hawaiian green sea turtles, J Wildl Dis
but without ocular tumors, turtles with less severe ocular 45:1138–1142, 2009.
tumors may be candidates for treatment if materials and 9. Patricio AR, Velez-Zuazo X, Diez CE, et al: Survival probability
trained personnel are available. In the same study, turtles of immature green turtles in two foraging grounds at Culebra,
with only flat, plaque-like FP lesions had a significantly Puerto Rico, Mar Ecol Prog Ser 440:217–227, 2011.
better prognosis, including spontaneous lesion regression 10. Patricio AR, Diez CE, van Dam RP: Spatial and temporal vari-
in more than 50% of cases, as compared with turtles with ability of immature green turtle abundance and somatic growth
in Puerto Rico, Endanger Species Res 23:49–60, 2014.
more verrucous-types of FP lesions.19 In general, FP is con-
11. Foley AM, Schroeder BA, Redlow AE, et al: Fibropapillomatosis
sidered more of an incidental finding in loggerhead turtles in stranded green turtles (Chelonia mydas) from the eastern United
(Caretta caretta), and rehabilitation efforts focused on log- States (1980-98): trends and associations with environmental
gerheads with FP may have a more favorable outcome than factors, J Wildl Dis 41:29–41, 2005.
those focused on green turtles with FP.20 Based on clini- 12. Aguirre AA, Balazs GH, Spraker TR, et al: Pathology of oropha-
cal findings, medical opinion, and permitting conditions, ryngeal fibropapillomatosis in green turtles (Chelonia mydas), J
the attending veterinarian should determine intake criteria, Aquat Anim Health 14:298–304, 2002.
euthanasia candidacy, and release criteria for turtles with FP. 13. Mader DR: Medical care of sea turtles: medicine and surgery. In
To be eligible for release, a turtle must be deemed capable Mader DR, editor: Reptile medicine and surgery, Philadelphia, PA,
of survival in its current condition. As long as the turtle 2006, Saunders, pp 977–1000.
is clinically stable and the overall assessment of survival 14. Herbst LH, Ene AR, Su M, et al: Tumor outbreaks in marine
turtles are not due to recent herpesvirus mutations, Curr Biol
following release is favorable, the presence of FP lesions
14:R697–R699, 2004.
should not prevent the release of rehabilitated turtles. It 15. Aguirre AA, Balazs GH, Zimmerman B, et al: Evaluation of
is recommended that green turtles be rehabilitated and Hawaiian green turtles (Chelonia mydas) for potential patho-
released as quickly as possible, because the stress of cap- gens associated with fibropapillomas, J Wildl Dis 30:8–15,
tivity may contribute to FP tumor development de novo 1994.
or exacerbate cycles of tumor removal and regrowth in 16. Tristan T, Shaver DJ, Kimbro J, et al: Identification of fibropapil-
ChHV5-infected turtles.19 Thus reduction of tumor burden lomatosis in green sea turtles (Chelonia mydas) on the Texas coast,
and rehabilitation to a clinically stable condition, including J Herp Med Surg 20:109–112, 2012.
treatment of any comorbid conditions, are acceptable goals 17. Van Houtan KS, Hargrove SK, Balazs GH: Land use, macroal-
in preparing a turtle for release. The risk of introducing gae, and a tumor-forming disease in marine turtles, PLoS ONE
ChHV5 into naive wild turtle aggregations via clinical or 5:e12900, 2010.
18. Work TM, Balazs GH, Rameyer RA, et al: Retrospective pathol-
subclinical carriers may be reduced by releasing rehabili-
ogy survey of green turtles (Chelonia mydas) with fibropapil-
tated turtles within the same geographic area from which lomatosis in the Hawaiian islands, 1993–2003, Dis Aquat Org
they were recovered. Marine turtle rehabilitation programs 62:163–176, 2004.
should take into account FP prevalence within local wild 19. Page-Karjian A, Norton TM, Krimer P, et al: Factors influenc-
marine turtle populations when evaluating resources and ing survivorship in rehabilitating green sea turtles (Chelonia
developing rehabilitation goals. mydas) with fibropapillomatosis, J Zoo Wildl Med 45:507–519,
2014.
References 20. Page-Karjian A, Norton TM, Harms C, et al: Case descriptions
of fibropapillomatosis in rehabilitating loggerhead sea turtles
1. Herbst LH: Fibropapillomatosis of marine turtles, Annu Rev Fish (Caretta caretta) in the southeastern United States, Dis Aquat
Dis 4:389–425, 1994. Organ 115:181–191, 2015.
402 SE C T I O N 11 Amphibians and Reptiles
21. Herbst LH, Jacobson ER, Moretti R, et al: Experimental trans- 38. dos Santos GR, Margins AS, Torezani E, et al: Relationship
mission of green turtle fibropapillomatosis using cell-free tumor between fibropapillomatosis and environmental quality: a case
extracts, Dis Aquat Organ 22:1–12, 1995. study with Chelonia mydas off Brazil, Dis Aquat Org 89:87–95,
22. Herbst LH, Moretti R, Brown T, et al: Sensitivity of the trans- 2010.
missible green turtle fibropapillomatosis agent to chloroform 39. Rodenbusch CR, Almeida LL, Marks FS, et al: Detection
and ultracentrifugation conditions, Dis Aquat Org 25:225–228, and characterization of fibropapilloma associated herpesvirus
1996. of marine turtles in Rio Grande do Sul, Brazil, Pesq Vet Bras
23. Whitley RJ: Herpesviruses. In Baron S, editor: Medical microbiol- 32:1179–1183, 2012.
ogy, ed 4, Galveston, 1996, University of Texas Medical Branch 40. Duarte A, Faisca P, Loureiro NS, et al: First histological and
of Galveston. virological report of fibropapilloma associated with herpesvirus
24. Work TM, Dagenais J, Weatherby TM, et al: In vitro replica- in Chelonia mydas at Principe Island, West Africa, Arch Virol
tion of chelonid herpesvirus 5 in organotypic skin cultures from 157:1155–1159, 2012.
Hawaiian green turtles (Chelonia mydas), J Virol 91:e404–e417, 41. Lackovich JK, Brown DR, Homer BL, et al: Association of
2017. herpesvirus with fibropapillomatosis of the green turtle (Chelonia
25. Page-Karjian A, Norton TM, Ritchie B, et al: Quantifying che- mydas) and the loggerhead turtle (Caretta caretta) in Florida, Dis
lonid fibropapilloma-associated herpesvirus in symptomatic and Aquat Org 37:89–97, 1999.
asymptomatic rehabilitating green sea turtles, Endanger Species 42. Quackenbush SL, Casey RN, Murcek RJ, et al: Quantitative
Res 28:135–146, 2015. analysis of herpesvirus sequences from normal tissue and
26. Work TM, Dagenais J, Balazs GH, et al: Dynamics of virus fibropapillomas of marine turtles with real-time PCR, Virol
shedding and in situ confirmation of chelonid herpesvirus 5 287:105–111, 2001.
in Hawaiian green turtles with fibropapillomatosis, Vet Pathol 43. Page-Karjian A, Torres F, Zhang J, et al: Presence of chelonid
52:1195–1201, 2015. fibropapilloma-associated herpesvirus in tumored and non-
27. Jacobson ER, Buergelt C, Williams B, et al: Herpesvirus in cuta- tumored green turtles, as detected by polymerase chain reac-
neous fibropapillomas of the green sea turtle (Chelonia mydas), tion, in endemic and non-endemic aggregations, Puerto Rico,
Dis Aquat Organ 12:1–6, 1991. Springerplus 1:35, 2012.
28. Herbst LH, Jacobson ER, Klein PA, et al: Comparative pathol- 44. Alfaro-Nunez A, Bertelsen MF, Bojesen AM, et al: Global distri-
ogy and pathogenesis of spontaneous and experimentally induced bution of chelonid fibropapilloma-associated herpesvirus among
fibropapillomas of green turtles (Chelonia mydas), Vet Pathol clinically healthy sea turtles, BMC Evol Biol 14:206–217, 2014.
36:551–564, 1999. 45. Herbst LH, Sundberg JP, Shultz LD, et al: Tumorigenicity of
29. Lu Y, Wang Y, Yu Q, et al: Detection of herpesviral sequences in green turtle fibropapilloma-derived fibroblast lines in immuno-
tissues of green turtles with fibropapilloma by polymerase chain deficient mice, Lab Anim Sci 48:162–167, 1998.
reaction, Arch Virol 145:1885–1893, 2000. 46. Herbst LH, Lemaire S, Ene AR, et al: Use of baculovirus-expressed
30. Greenblatt RJ, Work TM, Balazs GH, et al: The Ozobranchus glycoprotein H in an enzyme-linked immunosorbent assay
leech a candidate mechanical vector for the fibropapilloma- developed to assess exposure to chelonid fibropapillomatosis-
associated turtle herpesvirus found latently infecting skin tumors associated herpesvirus and its relationship to the prevalence
on Hawaiian green turtles (Chelonia mydas), Virol 321:101–110, of fibropapillomatosis in sea turtles, Clin Vaccine Immunol
2004. 15:843–851, 2008.
31. Kang KI, Torres-Velez FJ, Zhang J, et al: Localization of 47. Jones K, Ariel E, Gurgess G, et al: A review of fibropapillomatosis
fibropapilloma-associated turtle herpesvirus in green turtles in green turtles (Chelonia mydas), Vet J 212:48–57, 2016.
(Chelonia mydas) by in-situ hybridization, J Comp Pathol 48. Work TM, Balazs GH: Relating tumor score to hematology in
139:218–225, 2008. green turtles with fibropapillomatosis, J Wildl Dis 35:804–807,
32. Adnyana W, Ladds PW, Blair D: Observations of fibropapil- 1999.
lomatosis in green turtles (Chelonia mydas) in Indonesia, Aust 49. Norton TM, Jacobson ER, Sundberg JP: Cutaneous fibropapil-
Vet J 10:737–742, 1997. lomas and renal myxofibroma in a green turtle (Chelonia mydas),
33. Williams EH, Bunkley-Willams L, Peters EC, et al: An epizootic J Wildl Dis 26:265–270, 1990.
of cutaneous fibropapillomas in green turtles (Chelonia mydas) 50. Aguirre AA, Balazs GH, Spraker TR, et al: Adrenal and hemato-
of the Caribbean: part of a panzootic?, J Aquat Anim Health logical responses to stress in juvenile green turtles (Chelonia mydas)
6:70–78, 1994. with and without fibropapillomas, Physiol Zool 68:831–854,
34. Ene A, Su M, Lemaire S, et al: Distribution of chelonid 1995.
fibropapillomatosis associated herpesvirus variants in Florida: 51. Aguirre AA, Balazs GH: Blood biochemistry values of green
genetic evidence for infection of turtles following recruitment turtles (Chelonia mydas) with and without fibropapillomatosis,
to neritic developmental habitats, J Wildl Dis 41:489–497, Comp Haemotol Int 10:132–137, 2000.
2005. 52. Cray C, Varella R, Bossart GD, et al: Altered in vitro immune
35. Chaloupka M, Work TM, Balazs GH, et al: Cause-specific tem- response in green turtles (Chelonia mydas) with fibropapilloma-
poral and spatial trends in green turtle strandings in Hawaiian tosis, J Zoo Wildl Med 32:436–440, 2001.
Archipelago (1982–2003), Mar Biol 154:887–898, 2008. 53. Work TM, Raymeyer RA, Balazs GH, et al: Immune status of
36. Lloyd-Smith JO, Cross PC, Briggs CJ, et al: Should we expect free-ranging green turtles with fibropapillomatosis from Hawaii,
population thresholds for wildlife disease?, Trends Ecol Evol J Wildl Dis 37:574–581, 2001.
(Amst) 20:511–519, 2005. 54. Croft LA, Gorham JP, Schaf SA, et al: Evaluation of magnetic
37. Herbst LH, Klein PA: Green turtle fibropapillomatosis: chal- resonance imaging for detection of internal tumors in green
lenges to assessing the role of environmental cofactors, Environ turtles with cutaneous fibropapillomatosis, J Am Vet Med Assoc
Health Persp 103:27–30, 1995. 225:1428–1435, 2004.
CHAPTER 57 Fibropapillomatosis in Marine Turtles 403
55. Straub J, Jurina K: Magnetic resonance imaging in chelonians, 59. Norton TM: Chelonian emergency critical care, Sem Av Exotic
Semin Avian Exotic Pet Med 10:181–186, 2001. Pet Med 14:106–130, 2005.
56. Monezi TA, Mehnert DU, de Moura EMM, et al: Chelonid 60. Guimaraes SM, Wanderley AV, Monteiro-Neto C, et al: Evi-
herpesvirus 5 in secretions and tumor tissues of green turtles dence of regression of fibropapillomas in juvenile green turtles
(Chelonia mydas) from southeastern Brazil: a ten-year study, Vet (Chelonia mydas) caught in Niteroi, southeast Brazil, Dis Aquat
Microbiol 186:150–156, 2016. Org 102:243–247, 2013.
57. Alfaro-Nunez A, Gilbert MT: Validation of a sensitive PCR 61. Hirama S, Ehrhart LM: Description, prevalence and severity of
assay for the detection of chelonid fibropapilloma-associated green turtle fibropapillomatosis in three developmental habitats
herpesvirus in latent turtle infections, J Virol Methods 206:38–41, on the east coast of Florida, Fla Sci 70:435–448, 2007.
2014. 62. Brunner CH, Dutra G, Silva CB, et al: Electrochemotherapy for
58. Origgi FC: Herpesvirus and tortoises. In Mader DR, editor: the treatment of fibropapillomas in Chelonia mydas, J Zoo Wildl
Reptile medicine and surgery, Philadelphia, 2006, Saunders, pp Med 45:213–218, 2014.
814–821.
58
Rehabilitation Medicine of
Confiscated Turtles
BONNIE L. RAPHAEL
404
CHAPTER 58 Rehabilitation Medicine of Confiscated Turtles 405
B C
• Figure 58.1 Conditions of transport of Chelonians. (A) Truck used for transport of approximately 8000
Philippine forest turtles (Siebenrockiella leytensis). (B) Close view of turtles with broken shells in the bottom
of the truck. (C) Truckload of confiscated Philippine forest turtles. (All photos courtesy Katala Foundation.)
repacking, to alternate triage centers so that personnel and versed in chelonian medicine, treatment, and surgery—is
supplies do not have to be sent to one site. Where only necessary; in some instances up to seven veterinarians and
one species is involved, the equipment and supplies can 100 husbandry and logistics personnel at a time have been
be tailored to the size and type of animal. Gavage tubes of needed. Veterinary technicians are essential for the myriad
appropriate size, specific tube-feeding formulas, needle and of tasks—from record keeping to pharmacy maintenance
syringe sizes, amounts of hydration fluids, total amounts to laboratory testing and treatments—that need to be
and dilutions of drugs, and mouth specula are only some performed. For large confiscations with high numbers of
of the size and taxa-dependent adjustments that should be predicted deaths, a pathologist is invaluable.
made. For drugs that are most commonly used, it is very
useful to have spreadsheets that calculate the milligrams Triage and Initial Care
and volumes based on weights so that they do not have
to be generated in the field. Hard copies of the protocols, When confiscated animals are first encountered by gov-
waterproofed, are always necessary in the field, so these ernmental authorities, nongovernmental organizations
should be easily available for packing. (NGOs), or individuals, the first tasks are to provide all
In terms of necessary personnel, for most large confisca- the animals with at least minimal environmental necessities.
tions, one or two logistics people are essential. They will The provision of heat or cooling depends, of course, on
be involved with transportation logistics, communication the surroundings. Often just provision of shade is enough
with and among all the personnel, food and lodging, data cooling in tropical conditions, whereas provision of heat
recording and compilation, ordering supplies, organizing may involve simply placing animals in partially sunny areas
teams, final reports, and liaising between veterinary teams protected from wind. Care must be taken to adjust condi-
and other support staff. At least one veterinarian—well tions depending on time of day or changing sunlight or
406 SE C T I O N 11 Amphibians and Reptiles
shade. As animals are unpacked from primary containers simple as index cards with summaries of medical records or
(truck beds, boxes, bags, etc.), a quick assessment of triage as complex as extensive electronic communications can be
category is done and animals are separated by species (Fig. used. Support personnel should be keeping and transcribing
58.2). Triage categories should begin as large groupings and the information, freeing up the veterinary and animal care
then over time be refined into more individualized records. staff to perform the medical care.
One of the challenges is keeping track of which animals In confiscations of greater than 50 animals, time and
are in which group, when follow-up treatments need to be resources usually dictate that the initial handling be per-
performed, and when animals change triage levels. In the formed efficiently. This often requires estimation of body
case of large numbers of animals, organization is critical weights, the treatment of animals that may not need it,
in order to provide the best possible care while minimiz- and less than complete emergency care for animals that
ing the amount of handling and stress on the animals. At could benefit from it. Years of dealing with confiscations
minimum, clipboards with papers divided into columns have revealed that the most important factors affecting
for identification (ID) number, species, triage level, weight, immediate survival are how long the animals have been in
antibiotic, fluids, and so on should be on hand and data transport/confinement or without food and water. Initially,
recorded. if dealing with large groups of animals, individualized
In cases where animals are being transported to other medicine is minimized.
facilities for long-term care, communication with receiving Short periods of captivity without extreme deprivation
facilities regarding treatment is important. Something as of food or water usually result in animals that need the
CHAPTER 58 Rehabilitation Medicine of Confiscated Turtles 407
TABLE
58.1 Chelonian Drugs and Dosages
Concentrations Route(s) of
Drug, Generic Available Administration Dosage Frequency Reference
Antibiotics
Amikacin 50 mg/mL, 250 mg/ SQ, IM, ICe Loading dose 5 mg/kg q48–72h 9
mL then 2.5–3 mg/kg
Ceftazidime 1 g, 5 g SQ, IM, ICe 20 mg/kg q72h 10
Clarithromycin Various PO 15 mg/kg q24–72h 11
Danofloxacin 180 mg/mL IM, SQ 6 mg/kg q24–48h 12
Enrofloxacin 5 mg/mL, 100 mg/ PO, SQ, IM 10 mg/kg PO, 5–10 mg/ q48h PO, q24h inj 13–15
mL kg SQ, IM
Oxytetracycline 50, 100, 200 mg/mL SQ, IM 40 mg/kg then 20 mg/kg q72h 16
Ticarcillin 3 g with 0.1 g IM 100 mg/kg q48h 17
clavulanic acid
Tulathromycin 100 mg/mL IM 5 mg/kg q5–7days 18
Analgesics
Meloxicam 5 mg/mL IM 0.2 mg/kg q24–48h for 4 19
treatments
Tramadol Various PO 5–10 mg/kg 19
Fluids
Reptile ringers 1 part LRS + 2 parts SQ, ICe 5–50 mL/kg q24–72h prn 19
2.5% Dextrose in
0.45% NaCl
LRS or other SQ, ICe 5–50 mL/kg q24–72h prn 19
balanced solution
2.5% Dextrose and SQ, ICe 5–50 mL/kg q24–72h prn 19
0.45% NaCl
0.9% NaCl SQ, ICe 5–50 mL/kg q24–72h prn 19
Anesthetics
Ketamine 100 mg/mL IM 10–60 mg/kg 19
Medetomidine Various IM 0.1–0.15 mg/kg 19
Ketamine/ IM 5–10 mg/kg 0.1–0.15 mg/ 19
medetomidine kg
Atipamizole 5 mg/mL IM 0.5–0.75 mg/kg 19
Alfaxalone 10 mg/mL IV, IM, ICe 6–9 mg/kg IV, 9–15 mg/ 19
kg IM 24 mg/kg ICe
Propofol 10 mg/mL IV 5–10 mg/kg 19
Dewormers
Levamasol Variable SQ, ICe 5 mg/kg q10–21d 19
Fenbendazole 100 mg/mL PO 10–25 mg/kg For 3 days or every 19
14 days for 3
treatments
Praziquantel 57 mg/mL PO, SQ, IM 5–8 mg/kg Repeat in 14 days 19
Metronidazole 5, 50, 100 mg/mL PO 25 mg/kg q24h for 5 days 19
or 50 mg/kg q14d
ICe, Intracoelomic; IM, intramuscular; IV, intravenous; PO, orally; SQ, subcutaneous.
408 SE C T I O N 11 Amphibians and Reptiles
TABLE
58.2 Chelonian Triage and Actions
B C
• Figure 58.2 Temporary holding conditions of confiscated turtles. (A) Turtles in small holding tubs
undergoing treatment. (B) Group holding pen for Philippine forest turtles (Siebenrockiella leytensis) ready
for release. (C) Individual holding for big-headed turtles (Platysternon megacephalum) in a large pool.
(Photos A and B courtesy Lisa Eidlin, Photo C courtesy Chris Hagen.)
CHAPTER 58 Rehabilitation Medicine of Confiscated Turtles 409
shortest and most straightforward periods of medical and in Table 58.2 have pharmacokinetic data to inform their use
conservatory care. Those animals are usually placed into the in turtles. Drugs with the longest interinjection intervals
level 1 triage enclosures; they often receive only the first include tulathromycin 5 mg/kg every 5–7 days, ceftazidime
triage treatments and then, after being placed in appropriate 20 mg/kg every 3 days, amikacin 5 mg/kg once and then
housing, begin feeding and drinking and require no further 2.5–5 mg/kg every 3 days, enrofloxacin 5–10 mg/kg every
care. If animals have been in the trade routes for longer 24–48 hours orally (PO), SQ, or intramuscularly (IM);
than a week, they will be starting to become dehydrated these are the usual initial treatments. Subsequent treatments
and should be provided with drinking or soaking water. are based on individual conditions, but consideration should
Sometimes this will be in the form of shallow pools or be given to at least following through a 7- to 10-day course
low-rimmed pans, in the case of terrestrial turtles; for others of treatment to minimize the development of bacterial
it may mean pools or tubs of 2–18 in. deep. resistance.
Animals that have been in the trade routes for more
than a week will often be in triage level 2 or 3. Treatment Endoparasitism
during the first few days includes not only antibiotics, but
subcutaneous (SQ) or intracoelomic (ICe) fluids and also Normal parasite loads in most turtles will be increased to
sometimes steroids and analgesics. Treatment of infected pathologic levels when stressors are applied. In conditions
eyes, wounds, and shell damage begins and requires ongoing where species are mixed together, it is also possible for
care for varying amounts of time. turtles that are naive to some parasites to become infected
Choice of antibiotics depends not only on pharmacoki- from other species. Superinfections of enteric parasites have
netic data but also on the advantage of treating only every been seen subsequent to both conditions. Treatment with
3 days with a cephalosporin versus daily injections of a injectable levamisole has been employed successfully in a
fluoroquinolone and penicillin. There is a temptation to use wide range of turtles. Fenbendazole has also been used
drugs known for being long-acting in birds or mammals on frequently, although caution should dictate that low, infre-
the assumption that the properties will be similar in reptiles. quent doses be used in order to avoid complications such
It is advisable, if at all possible, to start antibiotic treat- as bone marrow suppression. Under no conditions should
ment with drugs for which there is some level of chelonian ivermectin be used. Fecal flotations should be performed
pharmacokinetic information. to evaluate what type of parasite is present and so that
subsequent dewormings may be adjusted. Praziquantal
Syndromes may be used to treat trematode infections. In the case of
amoebae or high numbers of flagellated protozoans detected
The syndromes that are commonly seen in confiscated via microscopic examination of fecal or cloacal swab smears,
turtles may involve various body systems. In the immediacy metronidazole should be given via gavage.
of a confiscation, treatments are based on what can be
done efficiently with the highest probability of relieving Starvation
life-threatening conditions. Conditions that are not urgent
should be deferred until all animals have been assessed and Many animals will be extremely thin, their fat stores having
undergone basic care. been used up, often bordering on terminal starvation. Intro-
duction to calories must be done in a slow, measured manner
Dehydration in order to avoid inducing the complication of refeeding
syndrome. Slowly increase the amount being gavage-fed
If “reptile Ringers” (see Table 58.1) is available, it is pre- over 1–2 weeks until a quantity of approximately 10 mL/kg
ferred over other fluids, although standard prepackaged body weight can be administered. Fluid support will need
fluids (i.e., 2.5% dextrose in 0.45% NaCl) have been used to be continued during that time. Although gavage feeding
successfully.7 Fluids may be administered SQ, although the may be safely repeated, sometimes it is less stressful on the
ICe route will correct dehydration more rapidly. Doses of animals to place a pharyngostomy tube surgically. These
10–50 mL/kg are tolerated and can be repeated daily if tubes can be left in place until the animals start feeding
necessary. All animals should be provided with drinking voluntarily.
water in low bowls (to prevent drowning) as soon as is
practical. Syndromes Affecting Eyes
Septicemia, Bacterial Enteritis Blepharitis, conjunctivitis, and corneal ulcerations are all
seen (Fig. 58.3A). Stain corneas if possible; otherwise avoid
Most animals that have been in the trade or held in sub- treating with steroids. Consider treatment with topical,
standard conditions are susceptible to bacterial infections. systemic, and subconjunctival antibiotics. Parenteral anal-
Administration of antibiotics via injection at the same time gesics are often also indicated. In the case of aquatic turtles,
as fluids are given is recommended if there are signs of stress, blepharitis and conjunctivitis often resolve spontaneously
dehydration, starvation, or wounds. Most of the antibiotics when water quality is improved.
410 SE C T I O N 11 Amphibians and Reptiles
B C
• Figure 58.3 Ocular and shell lesions in confiscated Chelonians. (A) Corneal plaque and edema in
a Philippine forest turtle (Siebenrockiella leytensis). (B) and (C) Necrotic shell lesions in aquatic turtles
subjected to hard substrates and transport of turtles in overcrowded conditions in a truck. (Photos
courtesy Sheena Koeth.)
Syndromes Affecting Skin or Shell chronically ill and infected, so they are not candidates for
immediate surgical intervention. Shell fractures should be
Ulcerations, abscesses, amputations, and ectoparasites are temporarily stabilized after cleaning.
encountered (see Fig. 58.3B and C). Animals that have After the initial triage and treatments, the animal’s con-
been held on hard substrates or piled upon each other ditions and triage categories may change. All animals should
often have significant shell lesions consisting of damage to be accounted for daily and assessed for change in condition.
the subkeratinized bone. In extreme conditions there may As soon as is practical, animals in category 1 should be
be full-thickness necrosis through the shells. These lesions offered food. Records of interest in food, acceptance of
must be generously debrided and protected with appropri- food, force-feeding, and amounts offered should be kept
ate ointments and the animals then placed on soft substrates if at all possible. Animals in the other categories may be
or in water in which they can float (if strong enough not offered food, although the frequency of treatments may not
to drown). Collect ectoparasites in alcohol; cleanse the be conducive to feeding responses.
wounds with dilute chlorhexidine, povidone-iodine (Beta- Ongoing treatments will be dictated by the conditions of
dine), or just soap and water. Debride or open and remove the animals, laboratory results and necropsy findings. Fecal
purulent material; flush copiously with sterile water and examinations (direct and flotations) should be performed on
dilute povidone-iodine. Apply ointment. Allow to maintain subsets of animals of each species and category. Necropsies
contact for 15 minutes if possible before returning to water. should be performed on as many animals of each species
Usually amputations of limbs are for those that have been as is reasonable.
CHAPTER 58 Rehabilitation Medicine of Confiscated Turtles 411
412
CHAPTER 59 Medical Evaluation of Crocodilians 413
TABLE
59.1 Common Clinical Signs and Differential Diagnosis
appearance but is not an accurate marker of the body condi- multiple pathogens are generally present.5,32–34 Chronic
tion. That storage fat is poorly vascularized and does not stress and poor hygiene are determining factors in the
mobilize rapidly in response to starvation.5,30 Crocodiles also occurrence of skin diseases (see Table 59.1).
possess an intracoelomic well-vascularized sack of readily
available adipose tissue, known as the steatotheca or fat Immune System
body.5,30 The steatotheca mobilizes promptly in response to
metabolic requirements; thus measurement of the fat body Ascertaining the immune status of the patient is a basic
is a more accurate score of the metabolic status of crocodil- function of the clinical examination. Crocodiles do not
ians.5 A steatotheca to heart ventricle (S:V) mass ratio of possess lymph nodes. Three lymphoid organs are acces-
5 or greater indicates an excess energy store. An S:V ratio sible to the clinician—the tonsils, spleen, and thymus. The
lower than 0.5 indicates very poor body condition.5 The tonsils consist of lymphoid tissue nested within mucosal
steatotheca is visible by ultrasound against the abdominal folds behind the soft palate.5,35 The tonsils can be visualized
wall of the right flank. The volume or maximum dimen- by lifting the soft palate and using an angled mirror or
sions of the steatotheca are compared with the ventricular telescope. Tonsils become reactive in a number of infec-
measurements to estimate the state of nutrition, using the tious diseases, including chlamydiosis and herpesvirosis,
previous ratio (Fig. 59.2).5,31 two significant epidemic diseases of hatchlings.5,6,36–38 The
spleen may be appraised by ultrasound from the right flank
Integumentary System beyond the fat body and below the gallbladder or from the
ventral window below and to the right of the gallbladder. It
Healthy crocodile skin is smooth, shiny, and dry. Scale is a well-defined oval organ with a homogeneously granular
morphology and the distribution of osteoderms and integu- texture. The spleen reacts to sepsis by enlarging, deforming
mentary sense organs (ISOs) vary between species. Cuvier (budding), and changing texture (see Fig. 59.2). The liver
dwarf caimans (Paleosuchus palpebrosus) are very heavily also provides important clues regarding the septic state of
armored with thick osteoderms in all scales, whereas estua- the patient (see later). Hematology and serum biochemistry
rine crocodiles (Crocodylus porosus) only have osteoderms in are of minor help, and at times misleading, in assessing the
the larger dorsal scales. ISOs in the alligators and caimans immune state of crocodiles and are secondary to clinical
(Alligatorinae) are confined to the head. Infectious skin findings including ultrasound. Immunocompromise is
diseases may present as macules, papules, crusts, or ulcers the result of chronic stress caused or aggravated by poor
and may be viral, bacterial, or fungal in origin, although management and is associated with moderate anemia, mild
CHAPTER 59 Medical Evaluation of Crocodilians 415
A1
B1
B2
A2
• Figure 59.2 Ultrasound and gross appearance of the spleen and steatotheca of Siamese croco-
dile (Crocodylus siamensis). (A1 and A2) Ultrasound and gross appearance of the spleen (white star)
with inflammatory deformity (circle). (B1 and B2) Ultrasound and gross appearance of a well-stocked
steatotheca (black star). (C) Depleted steatotheca (black star). Heart ventricle and spleen are shown for
reference.
hypophosphatemia, mild hypoalbuminemia, and increased septicemia. Pericardial and epicardial accretions of uric
serum corticosterone.5,22 Tonsillar, splenic, and thymic acid (visceral gout) may be diagnosed and sampled by
lymphoid populations are reduced in immunosuppressed ultrasound.
crocodiles.22
Respiratory System
Circulatory System
Crocodiles have two symmetrical saccular lungs, no air sacs,
The crocodile cardiovascular anatomy and physiology are and unidirectional air flow.42,43 Respiratory diseases often
probably the most sophisticated of all vertebrates.9,39–41 become evident only in advanced stages. “Open-mouth
Crocodilian hearts beat 30–50 beats per minute at behavior” is part of thermoregulation or social behavior and
rest on land and 5–8 beats per minute when diving.39 is not a sign of respiratory distress. Respiratory signs include
Heart rate decreases under anesthesia.40 Studies on reduced stamina, listing in the water, abnormal swimming,
crocodile clinical cardiology are lacking. Heart rate can foul smell on exhalation, nasal expulsion of stained exudate,
be counted by observing or sensing below the costal arch and gurgling rales. Angry animals may emit rumbling or
for cardiac movements. Ultrasound allows clear cardiac hissing noises. A water-soaked towel placed between the
visualization and measurements. Cardiac contractility of skin and the stethoscope facilitates auscultation. Percus-
anesthetized or moribund animals may be evaluated by sion of the lung field complements auscultation and may
ultrasound. Pericardial effusions occur in chlamydiosis or suffice to diagnose pulmonary consolidation or collapse.
416 SE C T I O N 11 Amphibians and Reptiles
Latero-lateral and dorso-ventral radiographs allow diagnosis On ultrasound the healthy yolk sac has a homogeneous
of obvious conditions in all species, but finer interpreta- appearance.
tion may be hampered by the outline of the osteoderms.
Computed tomography is the imaging tool of choice for the
evaluation of the crocodile respiratory system.44 Urinary System
Bacterial and fungal pneumonia typically present as
granulomatous focal or disseminated infections.5,44,45 Bron- Crocodiles excrete ammonia, uric acids, and small amounts
choscopy and bronchoalveolar lavages may be processed of urea. The crocodile kidney does not concentrate urine,
as in other species.46 Parasites are reported from lungs and water conservation takes place in the mucosal lining of
and pulmonary arteries.47 Pentastomidae infect crocodiles the distal intestines or in the urodeum.56,57 The kidneys are
through the consumption of fish and may cause verminous isoechoic with the fat they are encased in and are difficult to
pneumonia.48–51 identify on ultrasound. They may be visible from the pubic
window cranial to the urodeum or from the lateral windows
directing the beam ventral and cranial to the sacrum.
Digestive System Bacterial and parasitic nephritis are reported.5,47 In fasted
crocodiles, serum uric acid values higher than 12 mg/dL
Teeth are replaced continuously but progressively slower (750 µmol/L) accompanied by a calcium and phosphorus
with age. Diaphanous teeth may be a sign of stress-induced ratio less than 1 suggest renal disease. Gout is a common
osteoporosis.5 Nutritional bone disease is common in young manifestation of renal disease.5 In the winter, crocodiles are
animals and is diagnosed by pressing the two lower jaws predisposed to bouts of gout because urate crystals are less
together and bending the upper jaw upwards (rubber jaw).5 soluble at low temperatures and crocodiles are dehydrated
The stomach is located in the left flank and when full due to reduced feeding.58 Overfeeding may also cause gout.
may occupy much of the coelomic cavity, interfering with Urine may be obtained by inserting a semirigid catheter in
the ultrasound examination. On ultrasound, the stomach the cloaca past the proctodeum into the urodeum.59 Further
wall is thick with identifiable layers. Crocodiles routinely studies are needed to understand how urine composition
ingest stones and other foreign bodies. Gastric parasites may reveal systemic and organ changes. Urinary calcium is
are considered harmless, although studies linked gastric elevated in stressed crocodiles.5,22 Urine samples may also
parasites with histopathology findings.52,53 be used to investigate environmental pollutant metabolites
The duodenum is arranged in a double loop, and it and stress levels.60,61
and the pancreas located between its coils are visible on
ultrasound medial to the empty stomach. The intestines
are uniformly thick and widen when reaching the rectum. Reproductive System
On ultrasound the rectum can be recognized because it
abruptly ends where the hypoechoic urodeum (urinary Sexing is described earlier. The reproductive system of the
bladder) begins. Coccidiosis in juveniles may manifest as crocodile can be thoroughly assessed by ultrasound.62–64 In
a paradoxical obstipation due to accumulation of fibrin in hatchlings the minuscule paired gonads are nearly impos-
the lumen of the inflamed intestines.5 sible to locate. As the animals mature, the ovaries present
Normal feces are cylindrical, capped by white urates as clusters of anechoic or hypoechoic spheres of variable
within a varying degree of liquid urine. Fresh feces may size depending on reproductive status. Eggs are elongated
be obtained for parasitology and cytology examination by and calcified in crocodilians and easily distinguished from
digital stimulation of the cloaca or by rectal wash. Crocodil- follicles. Oophoritis presents on ultrasounds as clusters of
ian parasites have been catalogued.47 thick-walled heterogeneous cysts. Gestation or egg reten-
The large, symmetrical, bilobed liver envelops the heart tion can be diagnosed by radiographs. Estrogen and calcium
laterally and dorsally. Crocodiles have a gallbladder. Herpes serum levels in the Chinese alligator increase 20-fold and
virus, West Nile virus, chlamydiosis, and mycoplasmosis 3-fold, respectively, during follicular growth (Martelli,
cause severe diffuse hepatitis. During sepsis, the liver filters unpublished data). Reproductive success in females requires
circulating pathogens, causing focal or diffuse hepatitis. suitable males, adequate nesting sites and nesting material,
Therefore hepatitis may be a sequel and an indicator of tolerable intragender competition, and functional intact
sepsis (Fig. 59.3). Hepatitis is best diagnosed by ultrasound. ovaries and oviducts.
Changes within the hepatic parenchyma seen on ultrasound Testicles are elongated homogeneous paired organs
are verifiable on gross necropsy and histopathology (see visible on ultrasound from both flanks. Reproductive per-
Fig. 59.3).31 Hepatic neoplastic lesions are reported, but formance in males requires a peaceful disposition toward
neoplasia is rare in crocodilians,54 and expanding masses females and an intact penis. Semen can be collected by
are more often exuberant granulation tissue or abscesses.5,55 manually milking and stimulating the penis during the
In hatchlings and yearlings, the yolk sac occupies a large breeding season.65 Serum testosterone levels increase 30- to
part of the coelom. Omphalophlebitis and yolk sac infec- 380-fold during the mating season in the Chinese alligator
tions are associated with inadequate posthatching hygiene. (Martelli, unpublished data).
CHAPTER 59 Medical Evaluation of Crocodilians 417
B C
D E
A F G
• Figure 59.3 (A) Postmortem gross appearance of the liver of an adult wild male Nile crocodile
(Crocodylus niloticus) with extensive necrotizing hepatitis attributable to chronic sepsis of traumatic origin
from a tail injury. Ultrasound (B and C), gross (D and E), and histopathology (F and G) appearance of the
liver in two farmed Siamese crocodiles (Crocodylus siamensis).
B D
• Figure 59.4 Venipuncture sites in crocodilians. (A) Mandibular shelf in estuarine crocodile (Crocodylus
porosus), (B) mandibular shelf in Chinese alligator (Alligator sinensis), (C) dorsal venous sinus in (Croco-
dylus porosus), and (D) ventral tail vein in gharial (Gavialis gangeticus).
and trypanosomes, are common in crocodiles and do not 5. Huchzermeyer FW: Crocodiles: biology, husbandry and diseases,
appear to cause disease.49,75 An improvement in packed cell 2003, CABI Publishing.
volume (PCV), total protein, and albumin, in the absence 6. Youngprapakorn P, Ousavaplangchai L, Kanchanapangka S:
of leukopenia, is of positive prognostic value in patients A color atlas of diseases of the crocodile, 1994, Chulalongkorn
recovering from long-term debilitating conditions. University Press.
7. Divers SJ, Mader DR: Reptile medicine and surgery, ed 2, 2005,
Elsevier Health Science.
Acknowledgment 8. Jacobson ER: Infectious diseases and pathology of reptiles, 2007,
Taylor and Francis Group.
We thank Karthiyani Krishnasamy for valuable comments 9. Webb G, Manolis C: Australian crocodiles, 2007, Reed New
on the manuscript. Holland.
10. Molina FB: Class Reptilia, Order crocodilian: Caimans, croco-
References diles. In Biology, medicine, and surgery of South American wild
animals, Iowa State University Press. 2000, pp 9–13.
1. Neil WT: The last ruling reptiles, alligators, crocodiles and their kin, 11. Köhler G: Diseases of amphibians and reptiles, 2006, Krieger
1971, Columbia University Press, p xii. Publishing Company. English ed.
2. Webb GJW: Wildlife conservation: in the belly of the beast, 2013, 12. Jacobson ER, Behler JL, Jarchow JL: Health assessment of
Charles Darwin University Press, pp 121–158. chelonians and release into the wild. In Zoo and wild animal
3. Caldwell J: World trade in crocodilian skins 2011-2013, 2015, medicine current therapy (vol 4). 1999, pp 232–242.
UNEP-WCMC Cambridge. 13. Fleming GJ: Crocodilians (crocodiles, alligators, caiman, and
4. Zoological Information Management System. https://zims gharial). In Zoo animal and wildlife immobilization and anesthe-
.species360.org/Main.aspx. sia, ed 2, 2014, John Wiley & Sons, pp 325–336.
CHAPTER 59 Medical Evaluation of Crocodilians 419
14. Olsson A, Phalen D, Dart C: Preliminary studies of alfaxalone 32. Huchzermeyer FW, Wallace DB, Putterill JF, et al: Identifica-
for intravenous immobilization of juvenile captive estuarine tion and partial sequencing of a crocodile poxvirus associated
crocodiles (Crocodylus porosus) and Australian freshwater croco- with deeply penetrating skin lesions in farmed Nile crocodiles
diles (Crocodylus johnstoni) at optimal and selected sub-optimal (Crocodylus niloticus), Onderstepoort J Vet Res 76(3):311–316,
thermal zones, Vet Anaesth Analg 40(5):494–502, 2013. 2009.
15. Olsson A, Phalen D: Preliminary studies of chemical immo- 33. Jacobson ER: In Infectious diseases and pathology of reptiles. 2007,
bilization of captive juvenile estuarine (Crocodylus porosus) and Taylor and Francis Group, p 403.
Australian freshwater (Crocodylus johnstoni) crocodiles with 34. Buenviaje GN, Ladds PW, Martin Y: Pathology of skin diseases
medetomidine and reversal with atipamezole, Vet Anaesth Analg in crocodiles, Aust Vet J 76(5):357–363, 1998.
39(4):345–356, 2012. 35. Putteril JF, Soley JT: Surface features of the roof of the pharyngeal
16. Lloyd ML: Crocodilian anesthesia. In Zoo and wild animal cavity of the Nile crocodile (Crocodylus niloticus). Proceedings of
medicine current therapy (vol 4). 1999, pp 205–216. the Microscopy Society of Southern Africa 31, 2001, p 69.
17. Kock MD, Burroughs R: Chemical and physical restraint of 36. Hyndman TH, Shilton CM, Wellehan JF, Jr, et al: Molecular
wild animals. International Wildlife Services (Africa). 2014, pp identification of three novel herpesviruses found in Australian
258–260. farmed saltwater crocodiles (Crocodylus porosus) and Australian
18. Franklin CE, Davis BM, Peucker SK, et al: Comparison of stress captive freshwater crocodiles (Crocodylus johnstoni), Vet Microbiol
induced by manual restraint and immobilisation in the estuarine 181(3–4):183–189, 2015.
crocodile (Crocodylus porosus), J Exp Zoolog Part A Comp Exp Biol 37. Shilton CM, Jerrett IV, Davis S, et al: Diagnostic investigation of
298(2):86–92, 2003. new disease syndromes in farmed Australian saltwater crocodiles
19. Pfitzer S, Ganswindt A, Fosgate GT, et al: Capture of farmed (Crocodylus porosus) reveals associations with herpesviral infec-
Nile crocodiles (Crocodylus niloticus): comparison of physiological tion, J Vet Diagn Invest 28(3):279–290, 2016.
parameters after manual capture and after capture with electrical 38. Huchzermeyer FW, Langelet E, Putterill JF: An outbreak of chla-
stunning, Vet Rec 175(12):304, 2014. mydiosis in farmed Indopacific crocodiles (Crocodylus porosus),
20. Crocodile Specialist Group: Resources of the Veterinary Science J S Afr Vet Assoc 79(2):99–100, 2008.
Group. http://www.iucncsg.org/pages/Resources-of-Vet-Science 39. Axelsson M, Franklin CE: Elucidating the responses and role
-Group.html. of the cardiovascular system in crocodilians during diving: fifty
21. Strik NI, Alleman R, Harr KE: Circulating inflammatory cells. In years on from the work of C.G. Wilber, Comp Biochem Physiol
Jacobson ER, editor: In Infectious diseases and pathology of reptiles, A Mol Integr Physiol 160(1):1–8, 2011.
2007, Taylor and Francis Group, pp 167–218. 40. Stegmann GF, Williams CJ, Franklin C, et al: Long-term surgical
22. Shilton C, Brown GP, Chambers L, et al: Pathology of runting anaesthesia with isoflurane in human habituated Nile crocodiles,
in farmed saltwater crocodiles (Crocodylus porosus) in Australia, J S Afr Vet Assoc 88(0):e1–e6, 2017.
Vet Pathol 51(5):1022–1034, 2014. 41. Summers AP: Warm-hearted crocs, Nature 434:833–834,
23. Busk M, Overgaard J, Hicks JW, et al: Effects of feeding on 2005.
arterial blood gases in the American alligator (Alligator mississip- 42. Farmer CG: Similarity of crocodilian and avian lungs indicates
piensis), J Exp Biol 203(20):3117–3124, 2000. unidirectional flow is ancestral for archosaurs, Integr Comp Biol
24. Lance VA: Is regulation of aromatase expression in reptiles the 55(6):962–971, 2015.
key to understanding temperature dependent sex determination? 43. Sanders RK, Farmer CG: The pulmonary anatomy of Alligator
J Exp Zool A Ecol Genet Physiol 311(5):314–322, 2009. mississippiensis and its similarity to the avian respiratory system,
25. Yatsu R, Miyagawa S, Kohno S, et al: TRPV4 associates envi- Anat Rec (Hoboken) 295(4):699–714, 2012.
ronmental temperature and sex determination in the American 44. Hall NH, Conley K, Berry C, et al: Computed tomography of
alligator, Sci Rep 5:18581, 2015. granulomatous pneumonia with oxalosis in an American alligator
26. Langer S, Ternes K, Widmer D, et al: The first case of intersexual- (Alligator mississippiensis) associated with Metarhizium anisopliae
ity in an African dwarf crocodile (Osteolaemus tetraspis), Zoo Biol var anisopliae, J Zoo Wildl Med 42(4):700–708, 2011.
33(5):459–462, 2014. 45. Roh YS, Park H, Cho A, et al: Granulomatous pneumonia in
27. Murray CM, Easter M, Merchant M, et al: Methyltestosterone a captive freshwater crocodile (Crocodylus johnstoni) caused by
alters sex determination in the American alligator (Alligator mis- Mycobacterium szulgai, J Zoo Wildl Med 41(3):550–554, 2010.
sissippiensis), Gen Comp Endocrinol 236:63–69, 2016. 46. Lafortune M, Gobel T, Jacobson E, et al: Respiratory bronchos-
28. Tucker AD, Limpus CJ, McDonald KR, et al: Growth dynamics copy of subadult American alligator (Alligator mississippiensis)
of freshwater crocodiles (Crocodylus johnstoni) in the Lynd River, and tracheal wash evaluation, J Zoo Wildl Med 36(1):12–20,
Queensland, Aust J Zool 54:409–415, 2006. 2005.
29. Campos Z, Mourão G, Coutinho M, et al: Growth of Caiman 47. Tellez M: A checklist of host parasites interactions in the order
crocodilus yacare in the Brazilian Pantanal, PLoS ONE 9(2):89363, Crocodylia, 2013, University of California Press.
2014. 48. Ladds PW, Mangunwirjo H, Sebayang D, et al: Diseases in
30. Azeez OI, Myburgh JG, Meintjes RA, et al: Histomorphology, young farmed crocodiles in Irian Jaya, Vet Rec 136:121–124,
ultrastructure and fatty acid composition of the adipose tissue 1995.
in pansteatitis, the potentials in understanding the underlying 49. Jacobson ER: Parasites and parasitic diseases of reptiles. In Infec-
mechanism and diagnosis of pansteatitis in the Nile crocodile, tious diseases and pathology of reptiles, 2007, Taylor and Francis
Lipids Health Dis 16(1):47, 2017. Group, p 579.
31. Martelli P, Kot BCW: Ultrasound, a powerful tool for health 50. Junker K, Boomker J, Bolton LA: Pentastomid infections in Nile
assessment in crocodilians. Proceedings of the 21st working crocodiles (Crocodylus niloticus) in the Kruger National Park,
meeting of the Crocodile Specialist Group, Manila, Philippines. South Africa, with a description of the males of Alofia simpsoni,
2012, p 205. Onderstepoort J Vet Res 66(2):65–71, 1999.
420 SE C T I O N 11 Amphibians and Reptiles
51. Adams I, Isaza R, Greiner E: Fatal pentastomiasis in captive development in female American alligators, Alligator mississip-
African dwarf crocodile hatchlings (Osteolaemus tetraspis), J Zoo piensis, in southwest Louisiana, Gen Comp Endocrinol 162(3):
Wildl Med 32:500–502, 2001. 251–256, 2009.
52. Zhao JH, Wang S, Tu G, et al: Histopathology of gastric wall in 65. Johnston SD, Lever J, McLeod R, et al: Semen collection and
Chinese alligator (Alligator sinensis) infected with Ortleppascaris seminal characteristics of the Australian saltwater crocodile (Cro-
sinensis, Nutr Hosp 32(3):1180–1183, 2015. codylus porosus), Aquaculture (vol 422–423), 2014, pp 25–35.
53. Cardoso AM, de Souza AJ, Menezes RC, et al: Gastric lesions in 66. Ross JP, Hoenyfield DC, Brown SB, et al: Gizzard shad thi-
free-ranging black caimans (Melanosuchus niger) associated with aminase activity and its effects on the thiamin status of captive
Brevimulticaecum species, Vet Pathol 50(4):582–584, 2013. American alligators (Alligator mississippiensis), J Aquat Anim
54. Hill AG, Dennis MM, Pyne M: Squamous cell carcinoma with Health 21(4):239–248, 2009.
hepatic metastasis in a saltwater crocodile (Crocodylus porosus), 67. Honeyfield DC, Ross JP, Carbonneau DA, et al: Pathology,
Aust Vet J 94(3):83–86, 2016. physiologic parameters, tissue contaminants, and tissue thiamine
55. Huchzermeyer FW, Cooper JA: Fibriscess, not abscess, resulting in morbid and healthy central Florida adult American alligators
from a localized inflammatory response to infection in reptiles (Alligator mississippiensis), J Wildl Dis 44(2):280–294, 2008.
and birds, Vet Rec 147:515–517, 2000. 68. Ladds PW, Sims LD: Diseases in young farmed crocodiles in
56. Kuchel LJ, Franklin CE: Morphology of the cloaca in the Papua New Guinea, Aust Vet J 67:323–330, 1990.
estuarine crocodile, Crocodylus porosus, and its plastic response 69. Myburgh JG, Kirberger RM, Steyl JC, et al: The post-occipital
to salinity, J Morphol 245(2):168–176, 2000. spinal venous sinus of the Nile crocodile (Crocodylus niloticus):
57. Laverty G, Skadhauge E: Adaptive strategies for post-renal its anatomy and use for blood sample collection and intravenous
handling of urine in birds, Comp Biochem Physiol A Mol Integr infusions, J S Afr Vet Assoc 85(1):965, 2014.
Physiol 149(3):246–254, 2008. 70. Warner JK, Combrink X, Myburgh JG, et al: Blood lead con-
58. Neogi T, Chen C, Niu J, et al: Relation of temperature and centrations in free-ranging Nile crocodiles (Crocodylus niloticus)
humidity to the risk of recurrent gout attacks, Am J Epidemiol from South Africa, Ecotoxicology 25(5):950–958, 2016.
180(4):372–377, 2014. 71. Millan JM, Janmaat A, Richardson KC, et al: Reference ranges for
59. Myburgh JG, Huchzermeyer FW, Soley JT, et al: Technique for biochemical and haematological values in farmed saltwater croco-
the collection of clear urine from the Nile crocodile (Crocodylus dile (Crocodylus porosus) yearlings, Aust Vet J 75(11):814–817,
niloticus), J S Afr Vet Assoc 83(1):8, 2012. 1997.
60. Brock JW, Bell JM, Guillette LJ, Jr: Urinary phthalate metabolites 72. Lovely CJ, Pittman JM, Leslie AJ: Normal haematology and blood
in American alligators (Alligator mississippiensis) from selected biochemistry of wild Nile crocodiles (Crocodylus niloticus) in the
Florida wetlands, Arch Environ Contam Toxicol 71(1):1–6, 2016. Okavango Delta, Botswana, J S Afr Vet Assoc 78(3):137–144,
61. Ganswindt SB, Myburgh JG, Cameron EZ, et al: Non-invasive 2007.
assessment of adrenocortical function in captive Nile crocodiles 73. Rossini M, García G, Rojas J, et al: Hematologic and serum
(Crocodylus niloticus), Comp Biochem Physiol A Mol Integr Physiol biochemical reference values for the wild Spectacled Caiman
177:11–17, 2014. (Caiman crocodilus crocodilus), from the Venezuelan plains, Vet
62. Vac MH, Verdade LM, Mereilles CF, et al: Ultrasound evalu- Clin Pathol 40(3):374–379, 2011.
ation of the follicles development in adult female broad-nosed 74. Hamilton MT, Kupar CA, Kelley MD, et al: Blood and plasma
caiman (Cayman latirostris). In Proceedings of the 11th working biochemistry reference intervals for wild juvenile American
meeting of the Crocodile Specialist Group. IUCN – The World alligators (Alligator mississippiensis), J Wildl Dis 52(3):631–635,
Conservation Union. Gland, Switzerland, 1992, pp 176–183. 2016.
63. Tucker AD, Limpus CJ: Assessment of reproductive status in 75. Fermino BR, Paiva F, Soares P, et al: Field and experimental
Australian freshwater crocodiles (Crocodylus johnstoni) by ultra- evidence of a new caiman trypanosome species closely phyloge-
sound imaging, Copeia 4:851–857, 1997. netically related to fish trypanosomes and transmitted by leeches,
64. Lance VA, Rostal DC, Elsey RM, et al: Ultrasonography of Int J Parasitol Parasites Wildl 4(3):368–378, 2015.
reproductive structures and hormonal correlates of follicular
60
Reptile and Amphibian Analgesia
KURT K. SLADKY
421
422 SE C T I O N 11 Amphibians and Reptiles
stimulus. As with mammals, amphibians and reptiles endomorphin were found in the central nervous system of
have myelinated and unmyelinated afferent fibers running the African clawed frog (Xenopus laevis).21
together in sensory nerves: large myelinated A fibers (Aβ);
small myelinated A fibers (Aδ); and small unmyelinated C
fibers (C).1,7 In amphibians, small, slowly conducting fibers Measurement and Quantification of Pain
(Aβ and C) transmit the majority of all impulses induced and Analgesia in Amphibians and Reptiles
by noxious heat, pinching, pinpricks, and the application
of dilute acetic acid to the skin.7 In addition, a multitude Measuring pain in any species, particularly amphibians and
of biochemical mediators (e.g., substance P, catecholamines, reptiles, is the most difficult hurdle in the study of pain
cytokines, prostaglandins [PGs], etc.) are involved in the and analgesic efficacy. In mammals, it is well established
sensation, signal transmission, and perception of pain and that perioperative pain management facilitates recovery and
or inflammation, which adds to the complexity associated healing, reduces morbidity and mortality, and contributes
with our understanding of nonmammalian pain.8,9 Like to more rapid return to normal behavior.4 An objective
mammals, amphibians and reptiles have all of the anatomic understanding of normal behavior of a particular species
structures considered critical for the recognition of pain: and the ability to differentiate the presence of abnormal
peripheral nociceptors, appropriate central nervous system behavior indicative of discomfort are critical to the study
structures and pathways, opioid receptors and endogenous of pain and analgesia. Methods for assessing and measuring
opioids, reduction of nociceptive response with analgesics pain in amphibians and reptiles have been described previ-
(although data are sparse), pain avoidance learning, and ously.4,7 Ideally, a combination of appropriate behavioral
suspension of normal behavior with pain.10 Recent research and physiologic parameters might best be employed to
in fish, amphibians, reptiles, and birds has demonstrated the measure pain and analgesia in amphibians and reptiles.
transmission of peripheral sensory signals via the spinal cord Along those same lines, the development of a species- and
to midbrain and forebrain regions that are homologous to context-specific ethogram for each species being evaluated
mammalian cortical and limbic structures (anterior dorsal would provide the best method for distinguishing normal
ventricular ridge and posterior dorsal ventricular ridge of from abnormal (e.g., painful) behaviors. Most commonly,
the pallium in amphibians and reptiles).1,5,11,12 Thus the animal pain or lack thereof is assessed before and after
physiologic and anatomic requirements for pain and anal- an invasive procedure, such as a surgical procedure. This
gesia appear to be remarkably similar among all vertebrate method requires the development of a behavioral etho-
species. gram, which, in turn, requires the observer to become well
versed in subtle behavioral differences through many hours
Opioid Receptors and Endogenous Opioids of observation and analysis (videotaped or live observa-
tion). Behaviors must be operationally defined, which will
The opioid receptor gene family is highly conserved across provide objectivity and reproducibility.4 For example, in our
multiple vertebrate orders (e.g., bovids, chickens, bullfrogs, laboratory we developed a behavioral ethogram to evalu-
and teleost and elasmobranch fishes).13 An extensive ate preoperative and postoperative behavioral responses to
published literature is available with respect to amphibian food intake, willingness to swim, and breathing in red-
opioid receptors in the central nervous system. Four opioid eared slider turtles (Trachemys scripta elegans) following a
receptors have been cloned and sequenced in the northern unilateral orchidectomy.22 In a study using ball pythons
leopard frog (Rana pipiens)14,15 and three main opioid (Python regius), feeding behavior was used as an indicator
receptors, µ, δ, and κ, were cloned and sequenced in the of pain associated with the administration of capsaicin.23
rough-skinned newt (Taricha granulosa).16 In an extension An alternative to studying postsurgical pain is to measure
of this work, the same laboratory compared binding of pain under strictly controlled laboratory conditions using
µ-opioid agonists to human brain µ-opioid receptors and established behavioral models, during which noxious stimuli
frog brain µ-opioid receptors and found that most µ-opioid (e.g., mechanical, thermal, and chemical) are applied to an
agonists have higher affinity for binding with the human anatomic location on the reptile subject.24,25
µ-receptor compared with the frog µ-receptor.17 In reptiles, The application of a noxious thermal stimulus provides
there is limited information on opioid receptors. In aquatic a well-established behavioral model for assessing pain and
turtles, µ- and δ-opioid receptors are located throughout analgesia in rodents.4 In our own studies, we have successfully
the brain, and δ-opioid receptors are more abundant than adapted this classic thermal nociception model developed
µ-opioid receptors.18 With respect to endogenous opioid- for use in rodents and determined that amphibians and
related neurotransmitters, proenkephalin-derived peptides reptiles show unambiguous, easily quantifiable withdrawal
are present in turtles with a distribution similar to that responses indistinguishable from those of rodents.24,25 This
in mammals and birds.19 In addition, the reptilian brain method has also been applied to amphibians.26 Although
(aquatic slider turtles, American alligators [Alligator mis- some might argue that a withdrawal response to a noxious
sissippiensis], and anole lizards) was found to contain large stimulus is not equivalent to pain, there is evidence in a
quantities of endogenous enkephalins, also known as endor- variety of species that noxious thermal stimulation caused
phins.20 In amphibians, β-endorphin, met-enkephalin, and molecular and cellular changes in the central nervous
CHAPTER 60 Reptile and Amphibian Analgesia 423
system.27 In amphibians, the majority of nociception of commonly used opioid drugs (see also Chapter 26).
and antinociception studies have relied on the use of the Tables 60.1 and 60.2 summarize commonly used analgesic
acetic acid wipe test, predominantly in northern leopard protocols in amphibians and reptiles.
frog, with some studies using African clawed frog.7,28 This Butorphanol tartrate, a κ-opioid agonist/µ-opioid
method has been used to test the efficacy of a variety of antagonist, was once considered the analgesic of choice in
opioid, nonopioid, and nonsteroidal anti-inflammatory reptiles. However, our laboratory demonstrated that this
drugs (NSAIDs).7 The primary problem associated with has no clear analgesic properties in red-eared slider turtles,
the acetic acid wipe test is that it causes inflammation in bearded dragons, corn snakes (Pantherophis guttatus), or
addition to being a noxious nociceptive chemical, which ball pythons.24,25 Consistent with our data, intramuscu-
is a confounding factor in attempts to discriminate only lar butorphanol (1 mg/kg) had no analgesic efficacy as
nociception. determined by use of a thermal noxious stimulus method
Physiologic changes have been used to quantify stress (Fleming, 2012)32a and no isoflurane-sparing effect in green
and pain in mammals, and this approach has been adapted iguanas (Iguana iguana).33 In ball pythons, butorphanol
for amphibians and reptiles as well.29,30 For example, heart administered at 5 mg/kg intramuscularly (IM) had no
rate significantly increased in ball pythons after subcutane- effect on physiologic parameters compared with saline.34
ous (SC) capsaicin administration.30 In this study, opioid Conversely, one study demonstrated that butorphanol
analgesic drugs did not alter this physiologic response. (1.5 and 8 mg/kg IM) provided analgesia in green iguanas
Recently, in a comparative anesthesia/analgesia study using exposed to a noxious electrical stimulus.35 In amphibians,
Oriental fire-bellied toads (Bombina orientalis), heart and eastern red-spotted newts (Notophthalmus viridescens) were
respiratory rates were depressed in toads immersed in described as showing more normal postoperative behavior
alfaxalone-butorphanol, but the same toads immersed in (e.g., feeding, body posture, and response to observer) after
alfaxalone-morphine had normal heart rates and depressed limb amputation if they were exposed to butorphanol via
respiratory rates.31 In another amphibian study, the number immersion bath (0.5 mg/L water) during recovery from
of eye blinks as well as gular respiration rate were used tricaine methansulfonate anesthesia.36 However, when
to measure pain associated with intracoelomic (ICe) butorphanol was added to alfaxalone for sedating oriental
administration of concentrated saline in Indian bullfrogs fire-bellied toads in an immersion bath, there was no anti-
(Rana tigrina).32 Changes in the resulting behaviors were nociceptive response to a mechanical noxious stimulus.31
then quantified following the administration of opioid On the other hand, morphine sulfate, a µ-opioid agonist,
drugs and NSAIDs in order to determine analgesic was demonstrated to be analgesic in amphibians and
efficacy. reptiles.4,7 In the previously mentioned oriental fire-bellied
toad study, there was a clear antinociceptive response to a
noxious mechanical stimulus when frogs were sedated with
Analgesic Drugs alfaxalone and morphine in an immersion bath.31 Using
Opioids and Opioid-Like Analgesics multiple experimental models—including the acetic acid
wipe test, the thermal noxious withdrawal stimulus test,
Opioids, the most effective drugs for controlling pain in and a mechanical noxious stimulus test (von Frey filaments)
mammals, are classified according to receptor subtypes—µ model of nociception—morphine, at very high doses
(mu), κ (kappa), and δ (delta). µ, δ, and κ opioid was antinociceptive in northern leopard frogs across all
receptors—abbreviated MOR, DOR, and KOR—mediate nociceptive models.37 Using the thermal noxious stimulus
the analgesic effects of opioids, whereas the role of the method, morphine was an effective analgesic in bearded
fourth type of opioid receptor, the nociceptin or orphanin dragons (1 and 5 mg/kg) and red-eared slider turtles (1.5
FQ receptor (ORL), is less clear.7 For pain management and 6.5 mg/kg), but data were not as clear in corn snakes
in mammals, many clinicians prefer administering either (even when morphine was administered at an extremely
a µ-opioid receptor agonist (e.g., morphine, fentanyl, high dose of 40 mg/kg.25 Similarly, morphine (5, 7.5, 10,
hydromorphone, etc.), a partial µ-opioid receptor agonist and 20 mg/kg) and pethidine (10, 20, and 50 mg/kg), a
(e.g., buprenorphine), or a mixed-opioid, κ-receptor synthetic short-acting µ-opioid agonist, provided analgesia
agonist–µ-receptor antagonist (e.g., butorphanol). In order in Speke’s hinged tortoises (Kinixys spekii) exposed to for-
for exogenous opioids to be effective analgesics in amphib- malin administered into a limb, which was reversible after
ians and reptiles, opioid receptors must be present. The gene naloxone administration.38 In further support of µ-opioid
family for opioid receptors (µ, κ, and δ) is highly conserved agonist analgesic efficacy in reptiles, morphine increased
across multiple vertebrate orders.13 Two snake species have limb withdrawal latencies in Nile crocodiles (Crocodylus
endogenous brain opiates, and red-eared slider turtles have niloticus)39 and increased tail flick latencies in anole lizards
both proencephalin-derived peptides and functional µ- and (Anolis carolinensis).40 Related to morphine, hydromor-
δ-opioid receptors in the brain.18,19 In the northern grass phone is a semisynthetic, µ-opioid receptor agonist that
frog, µ-, κ-, δ-, and the ORL opioid receptors were cloned.13 was demonstrated to provide analgesia in red-eared sliders
Although opioid receptors are expressed in amphibians and using the thermal hind limb withdrawal nociception model
reptiles, we are just beginning to understand the efficacy at 0.5 mg/kg SC for up to 24 hours.41
424 SE C T I O N 11 Amphibians and Reptiles
TABLE
60.1 Analgesic Protocols for Use in Amphibians
Dosage/
Concentration Route Frequency Comments References
Opioids
Buprenorphine 50–75 mg/kg ICe, SC NDA Resumption of normal behavior 36
after limb amputation, eastern
red-spotted newt
Butorphanol 0.2 mg/kg IM NDA No efficacy data, postoperative in 70
African clawed frog
25–33 mg/kg SC q8–12h Experimentally antinociceptive, 7
northern leopard frogs
Butorphanol/alfaxalone 0.5 mg/L Bath NDA Resumption of normal behavior 36
after limb amputation, eastern
red-spotted newt
0.5 mg/100 mL Bath NDA No evidence of analgesia in Oriental 31
fire-bellied toads
Fentanyl 0.5 mg/kg SC NDA Presurgical analgesia, American 71
bullfrog
0.8 mg/kg SC q24h Experimentally antinociceptive, 7, 14
northern leopard frogs
Methadone 10–300 nmol/g SC NDA Experimentally antinociceptive, 54
northern leopard frogs;
1000 nmol/g was fatal
Morphine 10–300 nmol/g SC q8–16h Experimentally antinociceptive, 54
northern leopard frogs;
1000 nmol/g was fatal
40.0–100.0 mg/kg SC NDA 7
10.0 mg/kg ICe NDA Antinociceptive in northern leopard 28
frogs; not analgesic in African
clawed frogs
10.0 mg/kg ICe NDA Analgesic in northern leopard frogs 72
Naloxone 0.01–10.0 mg/kg ICe NDA Antagonizes µ-opioid analgesia in 72
northern leopard frogs
Naltrexone 0.10–10.0 mg/kg ICe NDA Antagonizes µ-opioid analgesia in 72
northern leopard frogs
Oxymorphone 200.0 mg/kg IM q48h Analgesic in AAT in edible frogs 42
NSAIDs
Flunixin meglumine 25.0 mg/kg SC NDA Analgesia in African clawed frogs 28
and northern leopard frogs
Local Anesthetics
Lidocaine 2–4 mg/kg Topical, NDA No systematic data on local 4, 57
50 mg/kg SC analgesia; systemic effects at
high dose
Proparacaine Ophthalmic 1–2 drops Topical NDA Effective for intraocular pressure 60
Solution (0.5%) Topical measurement
Other Anesthetics
Dexmedetomidine 0.1–3.0 nmol/g SC NDA Analgesia for at least 8 h even at 64
lower concentrations, northern
leopard frogs
Dexmedetomidine + 0.3 + 20 mg/100 mL Bath NDA Evidence of analgesic efficacy but 31, 54
alfaxalone minimal sedation
AAT, Acetic acid test; h, hours; IM, intramuscular; ICe, intracoelomic; NDA, no data available; NSAIDs, nonsteroidal anti-inflammatory drugs; SC, subcutaneous.
CHAPTER 60 Reptile and Amphibian Analgesia 425
TABLE
60.2 Analgesic Protocols for Use in Reptiles
Continued
426 SE C T I O N 11 Amphibians and Reptiles
TABLE
60.2 Analgesic Protocols for Use in Reptiles—cont’d
IM, Intramuscular; IV, intravenous; IT, intrathecal; NDA, no data available; NSAIDs, nonsteroidal anti-inflammatory drugs; PO, orally; SC, subcutaneous; TC,
transcutaneous.
An analog of oxymorphone, oxymorphazone, a µ-opioid have been no systematic studies of oxymorphone efficacy
agonist, was demonstrated to be a relatively weak analgesic in any reptile species.
drug, even at 200 mg/kg, when subcutaneously admin- Fentanyl is a synthetic, µ-opioid receptor agonist
istered to edible frogs (Rana esculenta; now Pelophylax with 75–100 times the potency of morphine; it can be
esculentus) and subjected to the acetic acid wipe test, but it administered either transcutaneously as an impregnated
had an approximately 48-hour duration of effect.42 There patch, subcutaneously, intramuscularly, or intravenously.
CHAPTER 60 Reptile and Amphibian Analgesia 427
In amphibians, fentanyl and remifentanil (a similar drug can be administered orally and due to its relatively long
to fentanyl with twice the potency) were determined to duration of action. Tramadol and its major active metabo-
be antinociceptive in frogs after either SC or intraspinal lite, O-desmethyl-tramadol (M1), produce analgesia in
administration using the acetic acid wipe test.14 In fact, of mammals by activating µ-opioid receptors but also by
all opioid agonists tested in one study using northern grass inhibiting central serotonin and norepinephrine reuptake.46
frogs exposed to the acetic acid wipe test, fentanyl was Respiratory depression, a common side effect associated with
the most effective after SC administration.7 In two reptile administration of other µ-opioid agonists, is significantly
species, two separate studies have evaluated the pharma- diminished with tramadol administration in mammals.46,47
cokinetics of fentanyl. In ball pythons, fentanyl plasma In mammals, the analgesic effects of tramadol typically
concentrations reached 1 ng/mL within 4 hours of applica- begin within 30 minutes after oral administration, and last
tion of a transdermal fentanyl patch (12.5 µg/h)43 and were for 6 hours. In contrast, tramadol (5.0 mg/kg; oral [PO])
detectable by 4–6 hours and for greater than 72 hours in administered to turtles significantly increased withdrawal
the plasma of prehensile-tailed skinks (Corucia zebrata) (the latencies for 12–24 hours post-drug administration, and
fentanyl dose was applied at 10% exposure of total surface 6–96 hours after administration of the higher tramadol
area of a 25-µg/h patch for 72 hours).44 It remains unclear dosages (10 or 25 mg/kg PO or SC).48 In loggerhead turtles
whether there is any biological significance to these plasma (Caretta caretta), plasma concentrations of tramadol and M1
fentanyl concentrations. A recently completed study in our remained above the target concentration of ≥100 ng/mL for
laboratory evaluated the efficacy of transdermal fentanyl approximately 48 hours at a dose of 5 mg/kg PO and for 72
patch (12.5 µg/h) administration in ball pythons and found hours when tramadol was administered at 10 mg/kg PO.49
no analgesic efficacy, even after repeating the study.43 Our Subjectively, appetite, swimming, and general activity level
laboratory also confirmed that fentanyl was readily absorbed did not change after drug administration. Most human and
through the skin of the snakes and remained at very high nonhuman mammalian studies consider the tramadol target
plasma levels during patch application.43 In the same study, analgesic plasma concentration to be 100 ng/mL. Recently,
we determined that fentanyl patch application decreased tramadol pharmacokinetics and efficacy were evaluated in
respiration in the same snakes, so we know that fentanyl yellow-bellied slider turtles (Trachemys scripta scripta) after a
is biologically active in the snakes even though we cannot single intramuscular dose (10 mg/kg) administered in either
definitively determine analgesic efficacy. the hindlimb or forelimb.50 Using a thermal hindlimb with-
Buprenorphine is an effective analgesic in many mam- drawal latency test, antinociceptive efficacy appeared to last
malian species and is used extensively due to its longer dura- approximately 48 hours regardless of whether the tramadol
tion of action compared with other opioids. Buprenorphine was administered in the forelimb or hindlimb, and tramadol
has partial agonist activity at the µ-opioid receptor, partial and M1 remained above an atypically high target plasma
or full agonist activity at the δ-opioid receptor, and antago- concentration (1 µg/mL or 1000 ng/mL) for approximately
nist activity at the κ-opioid receptor. In eastern red-spotted 48 hours.50 Of interest was that the pharmacokinetic trends
newts subjected to limb amputation surgery, the amphib- were similar for tramadol administration in forelimbs and
ian subjects were described to show more normal postop- hindlimbs, but the concentrations of M1 was approximately
erative behavior (e.g., feeding, body posture, and response 20% higher in the plasma of the group receiving tramadol
to observer) after exposure to buprenorphine administered in the hindlimbs compared with those receiving tramadol in
intracoelomically compared with controls.36 After the acetic the forelimbs. The antinociceptive effects of tramadol have
acid wipe test was used in northern grass frogs, buprenor- not been studied systematically in any amphibian species,
phine was weakly analgesic compared with other µ-opioids, although our laboratory is currently evaluating the efficacy of
such as fentanyl.7 Buprenorphine pharmacokinetics in rep- tramadol in tree frogs using a thermal nociceptive stimulus.
tiles were determined after SC administration in red-eared With respect to deleterious side effects, respiratory
slider turtles; effective dosages ranged from 0.075 to 0.1 mg/ depression associated with tramadol administration in
kg, which provided plasma concentrations similar to those red-eared slider turtles was approximately 50% less than
associated with analgesic efficacy in humans for approxi- that measured after morphine administration.48 Therefore
mately 24 hours.45 Interestingly, plasma concentrations of tramadol appears to be a promising analgesic alternative to
buprenorphine were reduced by approximately 70% when traditional opioids in reptiles. As mentioned, one of the sig-
the drug was administered in the hind limb compared with nificant deleterious side effects associated with tramadol and
the forelimb, indicating a significant hepatic first-pass effect. all opioid drug administration in mammals is respiratory
The analgesic efficacy of buprenorphine has not been dem- depression. This problem is paralleled in reptile studies. Our
onstrated in reptiles. Buprenorphine did not alter responses laboratory determined that both butorphanol and morphine
to an electrical noxious stimulus in green iguanas.35 Sim- caused profound respiratory depression in turtles,24 whereas
ilarly, in our laboratory, buprenorphine (0.1, 0.2, and respiratory depression was significantly less when turtles
1.0 mg/kg SC) provided no analgesic efficacy in red-eared were administered tramadol. The bottom line is that it is
slider turtles exposed to a noxious thermal stimulus.41 imperative that clinicians continue to monitor respiration
Tramadol has become a widely used analgesic alterna- during and after procedures in which any opioid drugs are
tive to other opioids in veterinary medicine because it administered to reptile species.
428 SE C T I O N 11 Amphibians and Reptiles
Tapendatol, similar mechanistically to tramadol, is a still develop at the site of injury and will be transmitted to
human drug that shares µ-opioid receptor activation and the central nervous system after the effect of the block has
norepinephrine reuptake inhibition with tramadol, but ceased. Because of its significant analgesic effect, any local
tapendatol has only weak serotonergic reuptake and has block that is correctly executed will significantly decrease the
more potent opioid properties without an active metabolite. required amount of other anesthetic agents, but additional
Tapendatol was administered to red-eared and yellow-bellied analgesia is warranted for postoperative pain management
slider turtles in order to determine analgesic efficacy using a in certain cases. The limitations associated with local anes-
thermal noxious stimulus model and pharmacokinetics.51,52 thetic administration include a focal nervous block and
After intramuscular administration (5 mg/kg), tapendatol short duration of action. However, there is evidence in
plasma concentrations were detectable for approximately amphibians that lidocaine will have systemic effects at very
24 hours and the duration of antinociceptive effects was high doses.57 In American bullfrogs (Lithobates catesbeianus),
approximately 10 hours in both turtle species.51,52 The lidocaine (50 mg/kg), administered subcutaneously, caused
shorter duration of antinociceptive efficacy compared with a significant decrease in respiratory rate, a progressive loss of
tramadol may be due to the lack of an active metabolite. righting and palpebral reflexes, and contralateral toe pinch
Tapendatol has not been studied in amphibians. Although withdrawal. However, these systemic effects were not associ-
respiratory depression has not been investigated after tapen- ated with local analgesia; the frogs reacted to a forceps pinch
datol administration in reptiles, in humans it is thought to at the site of the lidocaine injection at the time of maximal
cause less respiratory depression compared with commonly systemic effects.57
administered µ-opioid agonist drugs. With respect to local effects of these anesthetics, there is
only one published reptile study in which mepivacaine was
Parenteral Anesthetics used as a mandibular nerve block in an American alligator.13
In this study, a nerve locator was used to facilitate the proce-
There are few published data demonstrating analgesic dure. Lidocaine (2%) (up to 5 mg/kg total dose) can be used
efficacy associated with administration of anesthetic drugs for local ring blocks or line blocks. Lidocaine may be diluted
such as ketamine, dexmedetomidine, medetomidine, mid- with bicarbonate or sterile water at a 1:1 ratio or greater to
azolam, or propofol in amphibians or reptiles. Whereas decrease the pain of injection and allow increased volume
α-2-adrenergic drugs are commonly used in combination to be infused without reaching a toxic dose. In addition,
with ketamine or midazolam for sedation in amphibians and topical lidocaine (4%) is commonly administered directly
reptiles, few data exist with respect to the analgesic effects of to wounds before and after debridement for local analgesia,
these drugs. Intraspinal dexmedetomidine was administered particularly in chelonian species. Intrathecal administration
to northern grass frogs, causing a dose-dependent increase of local anesthetics is useful for surgical procedures of the
in pain thresholds using the acetic acid wipe test.53 In a tail, phallus, cloaca, and hind limbs.58 Lidocaine (4 mg/kg,
similar study using the acetic acid wipe test in northern <1 hour duration), bupivacaine (1 mg/kg, 1–2 hours dura-
grass frogs, systemic administration of dexmedetomidine tion) or preservative-free morphine sulfate (0.1–0.2 mg/
was analgesic compared with other α-adrenergic agonists.54 kg, duration of up to 48 hours) can be administered
Using the acetic acid wipe test, a thermal noxious withdrawal intrathecally between the coccygeal vertebrae of turtles. For
stimulus test, and a mechanical noxious stimulus test (von example, bupivicaine (0.1 mL for each 10 cm of carapace)
Frey filaments) in northern leopard frogs, dexmedetomidine was administered intrathecally in order to facilitate surgical
was antinociceptive after exposure to the acetic acid and excision of fibropapillomas from the posterior flippers of
thermal noxious stimuli but not after exposure to the a green sea turtle (Chelonia mydas).4 In a different study,
von Frey filaments.55 A recently completed study in our lidocaine (1 mL/20–25 kg) was administered intrathecally
laboratory demonstrated that dexmedetomidine (0.1 mg/ in hybrid Galapagos tortoises prior to phallectomy surgery.59
kg) produced antinociception for up to 24 hours in ball For topical administration, proparacaine hydrochloride
pythons.56 Although there is interest in the fact that low- (0.5%) has been used clinically in amphibians and reptiles
dose ketamine provides analgesia in mammals, there are no during eye exams, especially in order to measure intraocular
data in amphibians or reptiles. pressure. In American bullfrogs, proparacaine was used to
block corneal sensation during a research project in which
Local Anesthetics as Analgesics intraocular pressures were measured for reference ranges.60
Proparacaine was demonstrated to be effective in block-
Local anesthetics can be used alone or as part of a multimodal ing corneal sensitivity in Kemp’s ridley turtles for up to
anesthetic/analgesic approach. These include lidocaine, 45 minutes, with a 1-minute onset to effect.61 Similarly,
bupivacaine, and mepivacaine, which block peripheral nerve intrathecal analgesia was reported in chelonians.58
transmission to the dorsal horn by inhibiting sodium influx
into the neurons and therefore blocking the nociceptive Nonsteroidal Anti-Inflammatory Drugs
signal from traveling along the nerve fibers. For all local
anesthetics, pain transmission is blocked as long as the local Although they are not as potent as the opioids, NSAIDs are
anesthetic nerve block lasts, but inflammation and pain will used widely in reptile clinical practice as analgesics as well as
CHAPTER 60 Reptile and Amphibian Analgesia 429
for their anti-inflammatory properties but less so in amphib- for approximately 8 hours (IM) or 12 hours (ICe).69
ians. NSAIDs provide analgesia in mammals by blocking Ketoprofen (2 mg/kg IV), administered to green iguanas,
the binding of arachidonic acid to cyclooxygenase enzyme had a long half-life (31 hours) compared with ketoprofen
(COX), preventing the conversion of thromboxane A2 to pharmacokinetics in mammals, but the bioavailability after
thromboxane B2 (TBX), thus preventing the production IM administration was 78% with a relatively short half-life
of PG, potent mediators of inflammation.62 In bullfrogs, (8.3 hours).70
administration of meloxicam at a dosage of 0.1 mg/kg once Because no efficacy data and few pharmacokinetic data
daily suppressed circulating serum PGE2 levels following a are available with respect to NSAID administration in
controlled injury (i.e., punch biopsy).63 NSAIDs are often amphibians and reptiles, appropriate dosages and frequency
classified based on their relative specificity. There are two of administration can only be extrapolated. In addition,
COX enzymes, COX-1 and COX-2, which participate in clinicians should be aware of the deleterious side effects
renal and gastric protection and inflammation, respectively.62 documented in avian and mammalian species (e.g., renal
The NSAID ketoprofen is equipotent against both isoen- impairment, gastrointestinal ulceration/inflammation,
zymes, whereas carprofen is slightly more COX-2–specific hematologic abnormalities); assessment of suitability for
and meloxicam is COX-2–specific.62 Therefore the degree this class of drugs for the individual patient is prudent.
of efficacy and side effects may vary with each of the three
NSAIDs. Multimodal Analgesic Approaches
Although many NSAIDs appear to be relatively safe
when used in amphibians and reptiles, there are only a few In amphibians and reptiles, multimodal drug paradigms
published studies with respect to analgesic efficacy: one may be the best approach for managing pain. Multimodal
study uses a reptile species and three studies use amphibian analgesia refers to the administration of multiple drugs
species.7,28,34 Ball pythons administered meloxicam (0.3 mg/ that have analgesic efficacy at different levels in the central
kg IM) prior to surgical placement of an arterial catheter and peripheral nervous system. For example, opioids will
showed no physiologic changes (e.g., heart rate, blood have greatest efficacy at opioid receptors in the central and
pressure, plasma epinephrine, and cortisol) indicative of peripheral nervous system, whereas NSAIDs administered
analgesia. In leopard frogs undergoing the acetic acid wipe at the same time as the opioid will have greatest efficacy as
test, flunixin meglumine, a nonselective COX inhibitor, anti-inflammatory agents at the peripheral tissues. Local
provided antinociception after ICe administration, and the anesthetics can enhance multimodal analgesic protocols by
results were comparable to morphine administered to the blocking the initial pain cascade at the peripheral level. In
same subjects.64 In a similar study, two nonselective COX concert, all of these drugs have the potential to minimize
inhibitors, indomethacin and ketorolac, provided weak the transmission of pain signals to the brain, especially when
antinociception in leopard frogs exposed to acetic acid.65 administered preemptively, before a potentially painful
Although there are no published pharmacokinetic studies procedure is established (Tables 60.1 and 60.2).
of NSAIDs in amphibian species, several reptile studies
are in the literature. In a study using the thermal noxious
stimulus withdrawal latency model, flunixin meglumine References
(25 mg/kg), administered in the dorsal lymph sac, provided
better analgesia than xylazine hydrochloride or meloxi- 1. Sneddon LU: Evolution of nociception in vertebrates: com-
cam.28 Plasma concentrations of meloxicam (0.2 mg/kg parative analysis of lower vertebrates, Brain Res Brain Res Rev
46:123–130, 2004.
PO) administered as a single dose to green iguanas were at
2. Rose JD: Anthropomorphism and ‘mental welfare’ of fishes, Dis
plasma concentrations considered analgesic in mammals, Aquat Organ 75:139–154, 2007.
and these concentrations were measurable out to 24 hours 3. Altman DG, Bland JM: Absence of evidence is not evidence of
postadministration.66 In loggerhead turtles, meloxicam absence, BMJ 311:485, 1995.
(0.1 mg/kg) administered both IM and intravenous (IV) 4. Sladky KK, Mans C: Analgesia. In Mader DR, Divers S, editors:
did not reach plasma concentrations consistent with anal- Current therapy in reptile medicine and surgery, ed 3, St Louis,
gesia in humans, horses, or dogs, and the half-life was MO, 2014, Elsevier-Saunders, pp 217–228.
short.67 In a pharmacokinetic study in which meloxicam 5. Smith ESJ, Lewin GR: Nociceptors: a phylogenetic view, J Comp
(0.2 mg/kg IM and IV) was administered to red-eared slider Physiol [A] 195:1089–1106, 2009.
turtles, the IM dose provided a therapeutic concentration 6. Spray DC: Pain and temperature receptors of anurans. In Llinas
range necessary for meloxicam to provide analgesic and R, Precht W, editors: Frog neurobiology: a handbook, Berlin
Heidelberg, 1976, Springer-Verlag GmbH, pp 607–628.
anti-inflammatory effects equivalent in mammals for
7. Stevens CW: Analgesia in amphibians: preclinical studies
approximately 48 hours.68 In a different study evaluating and clinical applications, Vet Clin North Am Exot Anim Pract
the pharmacokinetics of meloxicam (0.2 mg/kg PO, ICe, 14:33–44, 2011.
and IM) in red-eared slider turtles, only the ICe and IM 8. Inagaki S, Senba E, Shiosaka S, et al: Regional distribution
routes, but not the PO route provided mean blood con- of substance P-like immunoreactivity in the frog brain and
centrations of meloxicam that were above those considered spinal cord: Immunohistochemical analysis, J Comp Neurol
effective to induce anti-inflammatory effects in mammals 201:243–254, 1981.
430 SE C T I O N 11 Amphibians and Reptiles
9. Reiner A, Krause JE, Keyser KT, et al: The distribution of hydrochloride in African-clawed frogs (Xenopus laevis), J Am
substance P in turtle nervous system: A radioimmunoassay and Assoc Lab Anim Sci 50:355–360, 2011.
immunohistochemical study, J Comp Neurol 226:50–75, 1984. 29. Moorea IT, Jessop TS: Stress, reproduction, and adrenocortical
10. Bateson P: Assessment of pain in animals, Anim Behav 42: modulation in amphibians and reptiles, Horm Behav 43:9–47,
827–839, 1991. 2003.
11. Donkelaar HJT, de Boer-van Huizen R: A possible pain control 30. Williams CJA, James LE, Bertelsen MF, et al: Tachycardia in
system in a nonmammalian vertebrate (a lizard, Gekko gecko), response to remote capsaicin injection as a model for nociception
Neurosci Lett 83:65–70, 1987. in the ball python (Python regius), Vet Anaesth Analg 43:429–434,
12. Munoz A, Munoz M, Gonzalez A, et al: Spinal ascending path- 2015.
ways in amphibians: cells or origin and main targets, J Comp 31. Adami C, d’Ovidio D, Casoni D: Alfaxalone-butorphanol versus
Neurol 378:205–228, 1997. alfaxalone-morphine combination for immersion anaesthesia
13. Li X, Keith DE, Evans CJ: Multiple opioid receptor-like genes in oriental fire-bellied toads (Bombina orientalis), Lab Anim
are identified in diverse vertebrate phyla, FEBS Lett 397:25–29, 50:204–211, 2016.
1996. 32. Pathak S, Patil PR, Suresh H, et al: A modified method for
14. Stevens CW: Opioid research in amphibians: an alternative pain evaluating analgesic activity of drugs using Rana tigrina frog, Int
model yielding insights on the evolution of opioid receptors, J Basic Clin Pharmacol 3:496–500, 2014.
Brain Res Brain Res Rev 46:204–215, 2004. 32a. Fleming GJ, Robertson SA: Assessments of thermal antino-
15. Stevens CW, Brasel CM, Mohan S: Cloning and bioinformatics ciceptive effects of butorphanol and human observer effect on
of amphibian mu, delta, kappa, and nociceptin opioid receptors quantitative evaluation of analgesia in green iguanas (Iguana
expressed in brain tissue: evidence for opioid receptor divergence iguana), Am J Vet Res 73:1507–1511, 2012.
in mammals, Neurosci Lett 419:189–194, 2007. 33. Mosley CA, Dyson D, Smith DA: Minimum alveolar concen-
16. Bradford CS, Walthers EA, Stanley DJ, et al: Delta and mu tration of isoflurane in green iguanas and the effect of butor-
opioid receptors from the brain of a urodele amphibian, the phanol on minimum alveolar concentration, J Am Vet Med Assoc
rough-skinned newt Taricha granulosa: cloning, heterologous 222:1559–1564, 2003.
expression, and pharmacological characterization, Gen Comp 34. Olesen MG, Bertelsen MF, Perry SF, et al: Effects of preopera-
Endocrinol 146:275–290, 2006. tive administration of butorphanol or meloxicam on physiologic
17. Brasel CM, Sawyer GW, Stevens CW: A pharmacological responses to surgery in ball pythons, J Am Vet Med Assoc
comparison of the cloned frog and human mu opioid recep- 233:1883–1888, 2008.
tors reveals differences in opioid affinity and function, Eur J 35. Greenacre CB, Schumacher JP, Tacke G, et al: Comparative
Pharmacol 599:36–43, 2008. antinociception of morphine, butrophanol, and buprenorphine
18. Xia Y, Haddad GG: Major difference in the expression of delta- in the green iguana, Iguana iguana, using electrostimulation, J
and mu-opioid receptors between turtle and rat brain, J Comp Herp Med Surg 16:88–92, 2006.
Neurol 436:202–210, 2001. 36. Koeller C: Comparison of buprenorphine and butorphanol anal-
19. Reiner A: The distribution of proenkephalin-derived peptides in gesia in the eastern red-spotted newt (Notophthalmus viridescens),
the central nervous system of turtles, J Comp Neurol 259:65–91, J Am Assoc Lab Anim Sci 48:171–175, 2009.
1987. 37. Willenbring S, Stevens C: Thermal, mechanical and chemical
20. Lindberg I, White L: Reptilian enkephalins: implications for peripheral sensation in amphibians: opioid and adrenergic
the evolution of proenkephalin, Arch Biochem Biophys 245:1–7, effects, Life Sci 58:125–133, 1996.
1986. 38. Wambugu SN, Towett PK, Kiama SG, et al: Effects of opioids in
21. Calle M, Claassen IE, Veening JG, et al: Opioid peptides, CRF, the formalin test in the Speke’s hinged tortoise (Kinixy’s spekii),
and urocortin in cerebrospinal fluid-contacting neurons in J Vet Pharmacol Ther 33:347–351, 2010.
Xenopus laevis, Ann N Y Acad Sci 1040:249–252, 2005. 39. Kanui TI, Hole K: Morphine and pethidine antinociception in
22. Kinney M, Johnson SM, Sladky KK: Behavioral evaluation of the crocodile, J Vet Pharmacol Ther 15:101–103, 1992.
red-eared slider turtles (Trachemys scripta) administered either 40. Mauk MD, Olson RD, Lahoste GJ, et al: Tonic immobility
morphine or butorphanol following unilateral gonadectomy, J produces hyperalgesia and antagonizes morphine analgesia,
Herp Med Surg 21:54–62, 2011. Science 213:353–354, 1981.
23. James LE, Williams CJ, Bertelsen MF, et al: Evaluation of feeding 41. Mans C, Lahner LL, Baker BB, et al: Antinociceptive efficacy of
behavior as an indicator of pain in snakes, J Zoo Wildl Med buprenorphine and hydromorphone in red-eared slider turtles
48:196–199, 2017. (Trachemys scripta elegans), J Zoo Wildl Med 43:662–665, 2012.
24. Sladky KK, Miletic V, Paul-Murphy J, et al: Analgesic efficacy 42. Benyhe S, Hoffmann G, Varga E, et al: Effects of oxymorphazone
and respiratory effects of butorphanol and morphine in turtles, in frogs: Long-lasting antinociception in vivo, and apparently
J Am Vet Med Assoc 230:1356–1362, 2007. irreversible binding in vitro, Life Sci 44:1847–1857, 1989.
25. Sladky KK, Kinney ME, Johnson SM: Analgesic efficacy of 43. Kharbush R, Gutwillig A, Hartzler K, et al: Transdermal fentanyl
butorphanol and morphine in bearded dragons and corn snakes, in ball pythons (Python regius) does not provide antinociception
J Am Vet Med Assoc 233:267–273, 2008. and decreases breathing frequency despite high plasma fentanyl
26. Vachon P: Hargreaves does not evaluate nociception follow- levels and brain mu-opioid receptor expression similar to opioid-
ing a surgical laparotomy in Xenopus leavis frogs, Res Vet Sci responsive turtles, Am J Vet Res, 2017.
97:471–474, 2014. 44. Gamble KC: Plasma fentanyl concentrations achieved after
27. Basbaum AI, Bautista DM, Scherrer G, et al: Cellular and transdermal fentanyl patch application in prehensile-tailed skinks
molecular mechanisms of pain, Cell 139:267–284, 2009. (Corucia zebrata), J Herp Med Surg 18:81–85, 2008.
28. Coble DJ, Taylor DK, Mook DM: Analgesic effects of meloxi- 45. Kummrow MS, Tseng F, Hesse L, et al: Pharmacokinetics of
cam, morphine sulfate, flunixin meglumine, and xylazine buprenorphine after single-dose subcutaneous administration in
CHAPTER 60 Reptile and Amphibian Analgesia 431
red-eared sliders (Trachemys scripta elegans), J Zoo Wildl Med juvenile Kemp’s Ridley sea turtles (Lepidochelys kempii), J Herp
39:590–595, 2008. Med Surg 25:116–121, 2015.
46. Lewis KS, Han NH: Tramadol: A new centrally acting analgesic, 62. Budsberg SC: Nonsteroidal anti-inflammatory drugs. In Gaynor
Am J Health Syst Pharm 54:643–652, 1997. J, Muir WW, editors: Handbook of veterinary pain management,
47. Mastrocinque S, Fantoni DT: A comparison of preoperative tra- ed 2, Toronto, 2009, Mosby, pp 183–239.
madol and morphine for the control of early postoperative pain 63. Minter LJ, Clarke EO, Gjeltema JL, et al: Effects of intramus-
in canine ovariohysterectomy, Vet Anaesth Analg 30:220–228, cular meloxicam administration on prostaglandin E2 synthesis
2003. in the North American bullfrog (Rana catesbeiana), J Zoo Wildl
48. Baker BB, Sladky KK, Johnson SM: Evaluation of the analgesic Med 42:680–685, 2011.
effects of oral and subcutaneous tramadol administration in red- 64. Terril-Robb LA, Suckow MA, Grigdesby CF: Evaluation of the
eared slider turtles, J Am Vet Med Assoc 238:220–227, 2011. analgesic effects of butorphanol tartrate, xylazine hydrochloride,
49. Norton TM, Cox S, Nelson S, et al: Pharmacokinetics of trama- and flunixin meglumine in leopard frogs (Rana pipiens), Contemp
dol and o-desmethyltramadol in loggerhead sea turtles (Caretta Top Lab Anim Sci 35:54–56, 1996.
caretta), J Zoo Wildl Med 46:262–265, 2015. 65. Stevens CW, MacIver DN, Newman LC: Testing and comparison
50. Giorgi M, Salvadori M, De Vito V, et al: Pharmacokinetic/ of non-opioid analgesics in amphibians, Contemp Top Lab Anim
pharmacodynamics assessments of 10 mg/kg tramadol intramus- Sci 40:23–27, 2001.
cular injection in yellow-bellied slider turtles (Trachemys scripta 66. Divers SJ, Papich M, McBride M, et al: Pharmacokinetics of
scripta), J Vet Pharmacol Ther 38:488–496, 2015a. meloxicam following intravenous and oral administration in
51. Giorgi M, Lee H-K, Rota S, et al: Pharmacokinetic and phar- green iguanas (Iguana iguana), Am J Vet Res 71:1277–1283,
macodynamics assessments of tapentadol in yellow-bellied slider 2010.
turtles (Trachemys scripta scripta) after a single intramuscular 67. Lai OR, Di Bello A, Soloperto S, et al: Pharmacokinetic behavior
injection, Journ Exotic Pet Med 24:317–325, 2015b. of meloxicam in loggerhead sea turtles (Caretta caretta) after
52. Giorgi M, De Vito V, Owen H, et al: PK/PD evaluations of the intramuscular and intravenous administration, J Wildl Dis
novel atypical opioid tapentadol in red-eared slider turtles, Med 51:509–512, 2015.
Weter 70:530–535, 2014. 68. Di Salvo A, Giorgi M, Catanzaro A, et al: Pharmacokinetic
53. Stevens CW, Brenner GM: Spinal administration of adrenergic profiles of meloxicam in turtles (Trachemys scripta scripta) after
agents produces analgesia in amphibians, Eur J Pharmacol single oral, intracoelomic and intramuscular administrations, J
316:205–210, 1996. Vet Pharmacol Ther 39:102–105, 2015.
54. Brenner GM, Klopp AJ, Deason LL, et al: Analgesic potency of 69. Uney K, Altan F, Aboubakr M, et al: Pharmacokinetics of
alpha adrenergic agents after systemic administration in amphib- meloxicam in red-eared slider turtles (Trachemys scripta elegans)
ians, J Pharmacol Exp Ther 270:540–545, 1994. after single intravenous and intramuscular injections, Am J Vet
55. Willenbring S, Stevens CW: Thermal, mechanical and chemi- Res 77:439–444, 2016.
cal peripheral sensation in amphibians: opioid and adrenergic 70. Tuttle AD, Papich M, Lewbart GA, et al: Pharmacokinetics
effects, Life Sci 58:125–133, 1996. of ketoprofen in the green iguana (Iguana iguana) following
56. Bunke LG, Sladky KK, Johnson SM: Antinociceptive efficacy single intravenous and intramuscular injections, J Zoo Wildl Med
and respiratory effects of dexmedetomidine in ball pythons 37:567–570, 2006.
(Python regius), Am J Vet Res (in press). 71. Van Bonn W: Clinical technique: extra-articular surgical stifle
57. Williams CJA, Alstrup AKO, Bertelsen MF, et al: When local stabilization of an American bullfrog (Rana catesbeiana), J Exotic
anesthesia becomes universal: Pronounced systemic effects of Pet Med 18:36–39, 2009.
subcutaneous lidocaine in bullfrogs (Lithobates catesbeianus), 72. Suckow MA, Terril LA, Grigdesby CF, et al: Evaluation of
Comp Biochem Physiol A Mol Integr Physiol 209:41–46, 2017. hypothermia-induced analgesia and influence of opioid antago-
58. Mans C: Clinical technique: Intrathecal drug administration in nists in leopard frogs (Rana pipiens), Pharmacol Biochem Behav
turtles and tortoises, J Exotic Pet Med 23:67–70, 2014. 63:39–43, 1999.
59. Rivera S, Divers SJ, Knafo SE, et al: Sterilisation of hybrid 73. Wellehan JFX, Gunkel CI, Kledzik D, et al: Use of a nerve
Galapagos tortoises (Geochelone nigra) for island restoration. locator to facilitate administration of mandibular nerve blocks
Part 2: phallectomy of males under intrathecal anaesthesia with in crocodilians, J Zoo Wildl Med 37:405–408, 2006.
lidocaine, Vet Rec 168:78, 2011. 74. Ruiz T, Campos WNS, Peres TPS, et al: Intraocular pressure,
60. Cannizzo SA, Lewbart GA, Westermeyer HD: Intraocular pres- ultrasonographic and echobiometric findings of juvenile Yacare
sure in American bullfrogs (Rana catesbeiana) measured with caiman (Caiman yacare) eye, Vet Ophthalmol 18:40–45, 2015.
rebound and applanation tonometry, Vet Ophthalmol 2017, 75. Schuster EJ, Strueve J, Fehr MJ, et al: Measurement of intraocu-
doi:10.1111/vop.12463. [Epub ahead of print]. lar pressure in healthy unanesthetized inland bearded dragons
61. Gornik KR, Pirie CG, Marrion RM, et al: Baseline corneal (Pogona vitticeps), Am J Vet Res 76:494–499, 2015.
sensitivity and duration of action of proparacaine in rehabilitated
61
Medical Aspects of Giant Tortoise
Relocation in the Galápagos Islands
JOSEPH P. FLANAGAN AND WASHINGTON TAPIA
T History
he Galápagos Islands are known for their biological
uniqueness and as the stimulus for Darwin’s The
Origin of Species. The islands lie 1000 km west of the The Galápagos Islands were discovered in 1535. Estimates
coast of Ecuador and straddle the equator. Currently, 97% of tortoise populations at the time suggest there may have
of the land area is National Park and the Galápagos Marine been as many as 250,000 throughout the islands. There
Reserve covers an area of 133,000 square kilometers sur- was very little human influence in the first century after
rounding the islands.1 The majority of the native and endemic their discovery; however, from the 17th through the 19th
species of Galápagos are still found there. However, the giant century, tortoises were harvested by buccaneers, whalers,
tortoises (Chelonoidis spp.) suffered decimation as a result and fur seal hunters as a source of fresh meat during
of human influence. Three species of tortoises have gone their long ocean journeys; and in the early 20th century
extinct (Table 61.1) due to humans, and the total numbers tortoises were harvested by local populations for food and
of several other species are greatly reduced from historical oil to be exported to the South American mainland.3,4 The
levels. tortoise population reached a low point in the 1970s, when
Giant tortoises are a keystone species in the Galápa- populations were estimated to be between 8000 and 14,000
gos ecosystem. The Galápagos National Park Directorate individuals.5
(GNPD) oversees the conservation and restoration of the Taxonomy of Galápagos giant tortoises has been a topic
islands. Restoration of tortoises to all the islands where they of debate for almost 200 years. For purposes of conservation
historically occurred and restoring populations to historic management, 15 species are described based on morpho-
numbers is a current collaborative program of the GNPD, logic, geographic, and molecular factors. There are five
the Galápagos Conservancy, international scientists, and species from Isabela Island, two from Santa Cruz Island, and
other nongovernmental organizations (NGOs).2 one each from the islands of Santiago, Pinzon, Espanola,
Conservation actions for tortoises have evolved from San Cristobal, Fernandina, Floreana, Santa Fe, and Pinta.
simply releasing juvenile tortoises to habitat restoration Populations on the latter four islands are extinct6,7 (Fig.
through elimination of introduced species and the use of 61.1; see Table 61.1). Restoring tortoise populations to
surrogate tortoises to serve as environmental engineers to historical numbers, including those considered “extinct
maintain the delicate balance of this fragile environment. in the wild,” is occurring through a combination of in
Rebuilding tortoise populations involves four major situ management, breeding and rearing tortoises where
program activities: captive breeding and release, head- appropriate, and, on islands where the endemic tortoise
starting and release of hatchlings from wild nests, release species is extinct, through the use of an analog (closely
of sterilized hybrid adult animals, and repopulating related) species.2 Among the 15 species, there are two
islands with breeding populations of genetically selected general morphologies: “dome” (carapace has smooth and
surrogate species. Veterinary involvement in the giant rounded contour as seen from the side, typical of most
tortoise program is relatively recent. The tortoise program chelonian species) shaped and “saddleback” (front of the
was solidly founded on a basis of good biological science carapace is raised and looks like a Spanish saddle in side
and animal husbandry. As the program matured, addi- view). The dome-shaped animals are generally larger (up
tional recognition was given to the potential contribu- to 250 kg or more) and are found on islands with higher
tions of nutrition, pathology, health monitoring, disease elevation where the vegetation is denser, the dome shape
screening, and surgical and medical care of specific facilitating movement through thick vegetation, whereas
cases. the saddleback tortoises are smaller (males up to 100 kg)
432
TABLE
61.1 Galápagos Giant Tortoise (Chelonoidis spp.) Population Threats and Conservation Actions
Main Current/
Map Species Morphology Range Population Historical Threats Conservation Actions Comment
A Chelonoidis Saddleback Fernandina Extinct Volcanic eruption Potential future surveys Natural extinction
phantastica
B C. becki Mixed Wolf Volcano 5–10,000 Goats/livestock, Removal of introduced species Source of rare/extinct genotypes
genetic
contamination
C C. microphys Domed Darwin Volcano 2000 Goats/livestock Removal of introduced species
D C. vandenburghi Domed Alcedo Volcano 5–10,000 Goats/livestock Removal of introduced species
E C. guentheri Domed Sierra negra 300–500 Human predation, Education efforts, captive
introduced species breeding, removal of introduced
species
F C. vicini Domed Cerro Azul 400–600 Human predation, Education efforts, captive
introduced species breeding, removal of introduced
species
G C. abingdoni Saddleback Pinta Island Extinct Human predation, Introduce surrogate species Sterilized tortoises introduced as
goats ecologic engineers
H C. darwini Intermediate Santiago Island 500–700 Goats, pigs, donkeys, Removal of introduced species, Goats, pigs, and donkeys
black rats head-starting eradicated 2005
I C. ephippium Saddleback Pinzon Island 150–200 Black rats Removal of introduced species, Rats eradicated 2012
head-starting
J C. elephantopus Saddleback Floreana Island Extinct Human predation, Removal of introduced species,
introduced species surrogate species introduction
K C. nigrita Domed SW Santa Cruz 5000 Human predation, Education efforts, removal
Island introduced species of introduced species,
head-starting
L C. donfaustoi Domed Eastern Santa Cruz 100 Human predation, Education efforts, removal
Island introduced species of introduced species,
head-starting
M C. spp. Saddleback Santa Fe island Extinct Human predation Analog species introduction Espanola tortoises introduced as
environmental engineers
N C. chathamensis Saddleback San Cristobal Island 2000 Human predation Removal of introduced species Natural population recruitment
O C. hoodensis Saddleback Espanola Island 2000 Human predation Captive breeding Founding population 3.12; goats
eradicated 1970s
CHAPTER 61 Medical Aspects of Giant Tortoise Relocation in the Galápagos Islands
433
434 SE C T I O N 11 Amphibians and Reptiles
• Figure 61.1 Map of the Galápagos Islands. Letters indicate the names of the islands or volcanos that
are home to the 15 identified species of Galápagos giant tortoises (Chelonoidis spp.) and correspond
to Table 61.1.
and are found on more arid islands of lower elevation with This restricts tortoise access to freestanding water and affects
more open vegetation. The elevation of the front of the the microclimate essential to certain life stages. Restoration
carapace allows these animals to reach higher when foraging of giant tortoise populations is fundamentally dependent on
in dry vegetation. A few island populations are intermediate the eradication of introduced pest species; and removal of
between dome and saddleback shapes (see Table 61.1; Fig. hunting pressure through public education.10
61.2A, B).8 Tortoise populations on Southern Isabela (C. vicina, C.
Human predation had the earliest impact on wild tortoise guentheri), Pinzon (C. ephippium), Santiago (C. darwini),
populations through harvesting of up to 200,000 animals and San Cristobal (C. chathamensis) were severely reduced by
for food and oil in the 17th–20th centuries. Deliberate human depredation but were also impacted by introduced
introductions of livestock (cattle, goats, swine, horses, species. Human exploitation reduced the population on
donkeys) and escape or accidental introduction of dogs, Espanola (C. hoodensis) to 2.12 (2 males, 12 females) which
cats, Norway rats, (Rattus norvegicus), black (roof ) rats, were so disperse on the island that breeding had ceased.
(R. rattus), and house mice (Mus musculus) have continued These animals were gathered and moved into a captive
population pressures on many of the species in their natural breeding program on Santa Cruz Island in the 1960s. They
habitats.9 The impacts of introduced vertebrate species were joined by a lone male returned from the San Diego
on giant tortoises are both direct and indirect. Eggs and Zoo to Galápagos in 1973 to make a founder population
hatchlings are predated by rats, dogs, cats, and pigs. Cattle, of 3.12 animals.11
donkeys, horses, goats, and pigs damage nesting areas, In addition, invasive plants alter the distribution of natu-
destroy food sources, and destroy or diminish the canopy of rally occurring native food sources and may affect seasonal
vegetation that naturally captures fog and mist precipitation. migration and alter the habitat, making it unsuitable for
CHAPTER 61 Medical Aspects of Giant Tortoise Relocation in the Galápagos Islands 435
A B
• Figure 61.2 (A) Saddleback morphology of the Pinta Island Galápagos giant tortoise (Chelonoidis
abingdoni) “Lonesome George” prior to his death and extinction of the species on June 24, 2012. (B)
Dome-shape morphology of Santa Cruz Island Galápagos giant tortoise (Chelonoidis nigrita).
use.12 When essential parts of the home range of individuals Isabela Island volcanos of Sierra Negra and Cerro Azul
are unsuitable, there are frequently no other options avail- was worsened by ongoing human harvest and depredation
able to the individuals dependent on that habitat. by introduced species. A new breeding center was built,
and a founding population of breeding age animals was
Restoration of Giant Tortoise Populations brought into captivity, with animals segregated into separate
subpopulations from different areas of these two volcano
Scientists recognized the giant tortoises of Galápagos seemed populations.13
to be doomed as early as the early 1900s. Tortoise collection
expeditions were made from the early 1900s to the late 1950s Head-Starting
for ex situ management. Animals from various islands were
deposited in zoos throughout the United States, Europe, Hatchlings and hatching eggs are collected from nesting
and Australia, often without identification of the island areas on Pinzon, Santiago, and Santa Cruz islands. Histori-
source or population of the individuals placed into captive cally, each tortoise population’s known nesting sites were
collections. Successful breeding outside of Galápagos is visited every year at the time of expected hatching, all
relatively rare but occurs in zoos and the private sector.8 observed nests were manually excavated, and all hatchlings
The Galápagos National Park was established in 1959 and unhatched eggs were removed for transport to Santa
to preserve and restore the unique flora and fauna of the Cruz for rearing. Although this practice has been successful
archipelago. Early efforts were begun in the 1960s to restore in producing high numbers of young tortoises for later
the tortoise population of Pinzon Island, where the total repatriation, it may have resulted in overrepresentation of
population of adults was estimated to be approximately a narrower genetic cross section of the population because
200 individuals. No successful recruitment had occurred the timing and location of collection was consistent from
in nearly 100 years, due to predation of hatchlings by the year to year. Recently, the practice changed to visiting one
introduced black rat. Hatching eggs and hatchlings still in island population per year and attempting to locate a higher
the nest were collected and transported to Santa Cruz Island number of nests to capture a broader representation of the
for rearing at the Tortoise Rearing Center until they were genetic diversity in each population.
large enough to be considered “rat proof ” at approximately Nests are opened with hand digging and small tools to
5 years of age, when they were returned to suitable habitat prevent accidental damage to the nest’s contents. Movement
on Pinzon.4 of reptile eggs during late incubation is less likely to result
Other populations were identified for head-starting. in embryonic damage because the membranes are more
These were reduced by human depredation but had signifi- stable; however, attention must be given to keeping the
cant ongoing population impact from introduced mammals egg orientation during transport and later artificial incuba-
that destroyed nesting areas and predated juvenile tortoises. tion the same as it was in the nest.14 Unhatched eggs and
Eggs and hatchlings from Santiago Island and Santa Cruz hatchlings from excavated nests are transported in small
were collected and transferred to the rearing center on Santa plastic containers containing lightly moistened vermiculite
Cruz for rearing under human care before release when to maintain humidity and reduce the impact associated with
less likely to suffer attacks by introduced predators. In the transport. Eggs are carried to a road (Santa Cruz) or to a
1990s the depletion of tortoise populations on the Southern boat (Santiago, Pinzon) for transport to the rearing center.
436 SE C T I O N 11 Amphibians and Reptiles
Developing eggs are hatched in incubators and hatchlings Prior to release, each animal is examined. They should have
placed in a “dark box” on moist vermiculite substrate for a good body condition and be capable of negotiating rough
month. This mimics the time spent by hatchlings in the nest terrain to find food and shelter and must have reached a
absorbing their yolk sac before they would emerge when the size that is likely to survive the risks they will encounter in
rainy season begins. Embryonic mortality is encountered the wild. Their diet is restricted to only leafy items, lacking
when the incubation temperature is too high or too low seeds for at least 30 days prior to release. This prevents the
(<25°C or >33°C). Eggs incubated at lower temperatures inadvertent dispersal of seeds of plants from Santa Cruz
have longer incubation times. Causes of embryonic and Island to the site of repatriation.12
neonate mortality have not been thoroughly investigated. Release occurs in suitable habitat near the nesting zone
Observations include unresorbed and inflamed yolk sacs, from which hatchlings were collected (or in the case of
which are presumed to be infection related, but factors animals bred in captivity, at historical nesting zones for
could include incubation problems.14 that population) during the wet or rainy season to ensure
Hatchlings are reared in rodent-proof enclosures, out- the availability of food and water for the juveniles as they
doors, on a substrate of crushed lava with approximately acclimate to the wild environment.13,14
50% shade. Hiding shelters are available with adequate
space for all hatchlings in the enclosure. Water is available Ecosystem Restoration
at all times in a shallow container that has small rocks
so hatchlings have good footing to facilitate egress. Lava Giant tortoises on Wolf Volcano of Isabela Island demon-
rocks are placed in the enclosures to provide visual barriers strate the full spectrum of tortoise shell morphologies found
between individuals, and climbing structures that provide historically in the Galápagos Islands. The larger population,
exercise. The diet consists almost exclusively of the leaves found primarily at higher elevations, is a dome-type tortoise
of two species of locally grown plants: Arrowleaf elephant that reaches a size of up to 250 kg. At the lower elevations
ear (Xanthosoma sagittifolium) and coral bean (Erythrina on the western slope of the volcano, there is a diverse
smithiana), which are offered three times per week. At 11 2 to population that includes full saddleback morphologies as
2 years of age they are moved into a larger “preadaptation” well as animals showing intermediate shapes ranging to fully
enclosure, where they have a substrate of lava rocks and soil domed. Genetic analysis shows that animals demonstrating
with natural vegetation to provide shade. Here they have the saddleback morphology have genes matching museum
constant access to water and are able to interact with other specimens originating from Floreana and Pinta Islands.15
juveniles up to 5 years of age.14 Some of these animals show hybridization with the species
Poor growth and soft shells are seen during prolonged native to Wolf, and some show genetic ingression from
periods of cool, wet weather; or when structures or vegeta- other island populations. Certain individuals sampled
tive growth results in too much shade, reducing the animal’s during surveys in 2008 and 2015 have a high degree of
ability to thermoregulate. Problems are more frequently purity to those extinct species and have the potential to
seen toward the end of the cool season, which occurs July be used in a breeding program to produce tortoises for
through December. Resolution of these problems occurs reintroduction with the genetic material that evolved on
with time when shade is reduced, allowing for improved those islands.
basking and increased ambient temperatures. Bite wounds The GNPD intends to reintroduce tortoises where they
are seen when hatchlings have restricted access to food or have been driven to extinction, with the goal of restoring
minimal ability to avoid enclosure mates. ecosystems. Goat eradication programs throughout the
Individual hatchlings are identified using painted islands have removed a major threat to tortoise survival
numbers; the color of paint is chosen to indicate the popula- and to ecosystem health but left these habitats without a
tion of origin, and they are numbered sequentially as they large herbivore to control woody vegetation. Thick growth
hatch. As they grow, painted numbers are refreshed until of woody vegetation prevents the growth of cactus (Opuntia
juveniles are old enough for placement of a passive induced spp.), a species essential to healthy tortoise populations.
transponder (PIT) tag in the left hind leg, which will then Woody vegetation obstructs tortoise movement and shades
identify them permanently. Prior to release, each individual the ground, reducing opportunities for tortoises to feed
is branded using red-hot metal on the carapace. This is a and thermoregulate. Tortoises consume grasses and forbs,
readily visible mark that may be seen in the field, indicating disperse seeds, and disrupt soils and serve to maintain a
the animal has a PIT tag. In certain field-monitoring situa- healthy ecosystem balance.
tions, animals are branded on the carapace with a number Without this large herbivore, scrub grasslands change
and/or are marked with a numbering system that involves to woody forested habitat. Pinta, Floreana, and Santa Fe
placing V-shaped notches in the marginal scutes to form Islands have been effectively without tortoises for a century
numbers from 1 to 9999. These procedures are typically or more.16
performed without anesthesia or analgesia.14 To some extent, feral goats have controlled woody veg-
Animals are weighed and measured every 3 months in etation but at the same time threatened native and endemic
captivity. They are released when they have reached a curved vegetation. Feral goats have recently been removed from the
carapace length of 20 cm, at approximately 3–4 years of age. uninhabited islands, and these islands now need tortoises
CHAPTER 61 Medical Aspects of Giant Tortoise Relocation in the Galápagos Islands 437
• Figure 61.4 Endoscope-assisted oophorectomy in a Galápagos • Figure 61.6 Galápagos giant tortoise (Chelonoidis spp.) phallec-
giant tortoise (Chelonoidis spp.). tomy procedure showing placement of clamp and ligatures.
animals will not be able to contribute to the genetics of the 11. Jiménez-Uzcátegui G, Márquez C, Snell HL: CDF checklist of
introduced population. Galápagos reptiles. In Bungartz F, Herrera H, Jaramillo P, et al,
editors: Charles Darwin Foundation Galápagos species checklist,
Charles Darwin Foundation Puerto Ayora, 2016, Galápagos.
Summary http://darwinfoundation.org/datazone/checklists/vertebrates/
reptilia/. Last updated 05 October 2016.
Despite centuries of decimation at the hands of humans 12. Sadeghayobi E, Blake S, Wikelski M, et al: Digesta retention
and from the impact of introduced animals, the tortoise time in the Galápagos tortoise (Chelonoidis nigra), Comp
populations in Galápagos are increasing. It will be decades Biochem Physiol A Mol Integr Physiol 160(4):493–497, 2011,
before numbers will approach aboriginal levels, and that doi:10.1016/j.cbpa.2011.08.008.
will occur only with active management. Restoration of tor- 13. Cayot LJ, Tapia W: Reign of the giant tortoises: repopulating
toises is one step in the recovery of Galápagos ecosystems. ancestral islands. In De Roy T: Galápagos, preserving Darwin’s
Additional work must be done in the removal of introduced legacy, Auckland, New Zealand, 2009, David Bateman Ltd, pp
vertebrates, invertebrates, and plant species that present 198–205.
ongoing threats. 14. Marquez C, Cayot LJ, Rea S: La Crianza de Tortugas Gigantes
en Cautiverio: Un Manual Operativo, Quito, Ecuador, 1999,
Fundacion Charles Darwin Para last Islas Galápagos.
References 15. Russello MA, Poulakakis N, Gibbs JP, et al: DNA from the past
informs ex situ conservation for the future: an “extinct” species of
1. Parque Nacional Galápagos Directorate website. Galápagos
Galápagos Tortoise identified in captivity, PLoS ONE 5(1):e8683,
Marine Reserve. Available at: http://www.galapagos.gob.ec/.
2010. https://doi.org/10.1371/journal.pone.0008683.
2. Galápagos Conservancy Web Site. Giant tortoise restoration
16. Gibbs JP, Hunter EA, Shoemaker KT, et al: Demographic out-
initiative. Available at: https://www.galapagos.org/.
comes and ecosystem implications of Giant Tortoise reintroduc-
3. Caccone A, Powell J: Giant tortoises: mapping their genetic past
tion to Española Island, Galápagos, PLoS ONE 9(10):e110742,
and future. In De Roy, T: Galápagos, preserving Darwin’s legacy,
2014. https://doi.org/10.1371/journal.pone.0110742.
Auckland, New Zealand, 2009, David Bateman Ltd, pp 98–105.
17. Keirans JE, Hoogstraal H, Clifford CM: The amblyomma
4. Merlin G: Restoring the tortoise dynasty: the decline and recovery
(Acarma: Ixodidae) parasitic on Giant Tortoises (Reptilia:
of the Galápagos giant tortoise, Quito, Ecuador, 1999, Fundacion
Testudinidae) of the Galápagos Islands, Ann Entomol Soc Am
Charles Darwin Para last Islas Galápagos.
66(3):673–688, 1973.
5. Froyd CA, Coffey EED, Knapp WO, et al: The ecological
18. Deem S, Jiménez-Uzcátegui G, Ziemmeck F. CDF checklist of
consequences of megafaunal loss: giant tortoises and wetland
Galápagos pathogens and parasites. In Bungartz F, Herrera H,
biodiversity, Ecol Lett 17(2):144–154, 2014.
Jaramillo P, et al, editors: Charles Darwin Foundation Galápagos
6. Caccone A, Gibbs JP, Ketmaier V, et al: Origin and evo-
Species Checklist Charles Darwin Foundation, Puerto Ayora,
lutionary relationships of giant Galápagos tortoises, PNAS
2011, Galápagos. http://www.darwinfoundation.org/datazone/
96:13223–13228, 1999.
checklists/ecological-groups/pathogens-and-parasites/. Last
7. Poulakakis N, Edwards DL, Chiari Y, et al: Description of a
updated 13 April 2011.
New Galápagos Giant Tortoise Species (Chelonoidis; Testudines:
19. Hunter EA, Gibbs JP, Cayot LJ, et al: Equivalency of Galápagos
Testudinidae) from Cerro Fatal on Santa Cruz Island, PLoS ONE
Giant Tortoises used as ecological replacement species to restore
10(10):e0138779, 2015, doi:10.1371/journal.pone.0138779.
ecosystem functions, Cons Biol 27(4):701–709, 2013.
8. Pritchard PCH: The Galápagos tortoises: nomenclatural and survival
20. Knafo SE, Divers SJ, Rivera S, et al: Sterilisation of hybrid
status, Lunenburg, MA, 1996, Chelonian Research Foundation.
Galápagos tortoises (Geochelone nigra) for island restoration.
9. Merlin G: Restoring the tortoise dynasty: the decline and recovery
Part 1: endoscopic oophorectomy of females under ketamine-
of the Galápagos giant tortoise, Quito, Ecuador, 1999, Fundacion
medetomidine anaesthesia, Vet Rec 168:47, 2011.
Charles Darwin Para last Islas Galápagos.
21. Rivera S, Divers SJ, Knafo SE: Sterilisation of hybrid Galápagos
10. Campbell K: Project Isabela, ecosystem restoration through
tortoises (Geochelone nigra) for island restoration. Part 2: phal-
mega-eradication. In De Roy, T: Galápagos, preserving Darwin’s
lectomy of males under intrathecal anaesthesia with lidocaine,
legacy, Auckland, New Zealand, 2009, David Bateman Ltd, pp
Vet Rec 168–178, 2011.
206–212.
SECTION 12
Avian
62 Antifungals in Birds, 441
440
62
Antifungals in Birds
KATHRYN C. GAMBLE
D
ue to immunosuppression and reduced or lapsed compared with newer cholesterol and liposomal formula-
husbandry standards, fungal infections in birds tions.7,25,26 It is highly protein bound, but tissue distribu-
housed in managed care remain a frequent veteri- tion is good, although limited concentrations occur in
nary presentation.1 Systemic fungal infections with oxyphilic cerebrospinal fluid (CSF) and bone.1,7,26 Direct application
and thermophilic Aspergillus predominate in the respiratory of AmpB to the infected area as nebulization (NE) or
tract, due to spore inhalation from environmental fungal lavage improves its efficacy and reduces toxicity potential.7
overgrowth from damp, protein-rich substrates.2,3 Vague Metabolism is poorly understood for AmpB, but elimina-
signs of early infection may go undetected, establishing tion is biphasic, and nearly 15 days in mammals, although
chronic disease challenges of direct lesion management and much shorter in birds.1,26
requirement for long-term treatment.2,4 Similarly, the other Dose-dependent nephrotoxicity, as a result of renal
primary fungal pathogen of concern for avian patients, vasoconstriction and direct nephron effect, is lessened in
Candida, is associated with poor nutrition, antibiotic treat- birds due to their faster avian elimination rate.1,7,26 Slowly
ment, or hand-rearing juveniles.5 Other fungal pathogens administered parenteral AmpB (1–1.5 mg/kg intravenous
are more limited in scope but warrant pharmacologic treat- [IV] twice daily [BID] to three times daily [TID] 3–5
ment. It has been suggested that measurement of antifungal days) can be combined with other antifungals for weeks.1,2,26
concentration in target tissues may be more useful than Topical lavage (1–1.5 mg/kg BID) for lesion rinse, or NE
plasma concentrations.6 To achieve optimal treatment (1 mg/mL for 15 minutes BID to four times daily [QID]
success, and especially for dimorphic fungi, antifungal sensi- can be considered for 3–7 days.1,2 Even though poorly
tivity or concentration measurements should be considered soluble in water, AmpB should be diluted before admin-
at available sources (http://strl.uthscsa.edu/fungus/). istration by any route, but balanced electrolytes or saline
as the diluent will inactivate the drug.1,26 Protected from
Aspergillosis Treatment (Table 62.1) light, sterile water dilutions are stable for 24 hours at room
temperature; 1 week under refrigeration; or 30 days at −4°F
There are four core classes of antifungal drugs with activity (−20°C).26 Some Aspergillus spp. have elevated minimum
against Aspergillus; the polyenes, including amphotericin inhibitory concentrations (MIC) beyond typical targets
B (AmpB); triazoles, including itraconazole (ITRA), flu- (0.5–2 µg/mL), and increasing resistance is reported.1,25,26
conazole (FLU), and voriconazole (VRC); allylamines,
including terbinafine (TERB); and fluocytosine.7 Azoles
441
442 SE C T I O N 12 Avian
10 SIDPK,CD
Voriconazole 20 BID-TIDPK SUSP Gavage ± food Mallard 14
*-(crushed into); CAP, capsule; OJ, acidic solution (e.g., orange juice); PK, Pharmacokinetics; POW, oral powder; -SAL, saline; -SU, suspending agent;
SUSP, suspension; TAB, tablet; TC, tissue concentrations; -W, water.
demonstrated in birds.25,27 In general, the oral suspension success,11,12,28 although some acquired resistance recently
produced more rapid absorption, and higher bioavailability has been documented.1,13,27
when patients were fasted, whereas capsules had improved Routinely administered ITRA doses (5–10 mg/kg by
absorption with food intake.27 Commercial formulations mouth [PO] once or twice a day [SID-BID]) may be
contain proprietary cyclodextrin, which improves absorp- administered as loading doses of increased frequency or
tion, and has marginalized use of compounded product due high end of dose range, then maintained for weeks to
to both markedly reduced ITRA concentrations measured months until clinical resolution at the lower dose ranges
in these products and treatment failures.10,11,25–28 A com- and frequency.3,26 However, plasma ITRA concentrations
mercially available subcutaneous controlled release gel was demonstrate species variability even at the same doses and
assessed in mallards that produced undetectable plasma consistent feeding schedule. In particular, granivore and
concentration of ITRA.8 carnivore gastric acidity differences may play a role in degree
Despite extensive protein binding, and due to its extreme of absorption and maximum concentration achieved.12,13
lipophilicity, ITRA is well distributed throughout the body, In addition, potential ventricular koilin binding has been
although brain concentrations were lower than those of reported in granivores.12 Body fat composition was proposed
plasma.26,27 Its principal metabolite hydroxy-ITRA is active as the explanation for the lower plasma concentrations
and contributes markedly in some avian species.1,11,26–28 achieved in anseriformes at even much higher than standard
Metabolites are excreted through bile and urine, whereas doses (20 mg/kg).8 In similar dose studies, tissue ITRA
unmetabolized ITRA is excreted via bile.1,27 Without concentrations have been measured in several avian species.
loading doses, the long elimination half-life may protract Especially the lung and brain, where systemic aspergillosis
achievement of steady state.1,26,27 With a broad spectrum is most prevalent, repeated documentation of their lowest
of efficacy,26 ITRA remains a primary choice for avian concentrations for all body tissues were noted regardless of
aspergillosis treatment,3,28 with target plasma concentra- the dose, even where other tissues were more reflective of
tion of 0.25 µg/mL in humans associated with clinical or exceeded plasma concentrations.1,8,9,13,27
CHAPTER 62 Antifungals in Birds 443
Of species-specific note, ITRA is not recommended in avian species, this actual parameter is quite variable
for African grey parrots (Psittacus erithacus timneh) due to between taxa.
frequent patient inappetance and depression and some- Even with standard avian dose recommendation
times death, although reduced oral dose recommendations (12.5 mg/kg PO BID × 60–90 days), and as compared
(2.5–5 mg/kg PO SID) have been published.1,6,26 Raptors with all other azoles, VRC has the most nonlinear pharma-
also may present significant anorexia.27 However, generally cokinetics and metabolic saturation, although essentially no
minimal impact to clinical pathology or hepatic function active metabolites are produced.3,14,21,28,31 This characteristic
is noted in birds. markedly minimizes interspecies extrapolation of doses,
or limits interpretation of extended regimens from single
Fluconazole dose or short-term studies.18,19,31 Autoinduction of its own
This synthetic triazole specifically inhibits demethylation metabolism is sufficient to require intraindividual adjust-
of lanosterol to ergosterol.28 Because it is highly water ments in long-term treatment regimens.1,17,19–21,26,28,31,34
soluble and has low protein binding, FLU is well absorbed When compounded from tablet formulation into water or
orally, unaffected by feeding or gastric pH.1,7,26 It exten- commercial suspending agent, VRC has been confirmed as
sively penetrates to CSF,25,26 although it has better effects stable under refrigeration for 14 days.18,26,35
outside the central nervous system.1,28 In contrast to all Primarily used for Aspergillus spp. treatment, VRC
other azoles, FLU is metabolized minimally and excreted targeted to MIC of ≤0.38–≤1 µg/mL is very effective,18,31
essentially unchanged by the kidney; therefore caution with but it is time, rather than concentration, dependent.21 It
renal insufficiency should be exercised.7,25,26,28 Compounded has shown little resistance to date and is increasingly used
oral suspension from commercially available tablets crushed as front line treatment not only for aspergillosis, but also
with water and commercial suspending agents have been Candida sp. and dimorphic fungi, but not for zygomy-
shown stable for 14 days when light protected and stored at cetes.3,28,31 Controlled clinical VRC treatment of Falco
41°F (5°C).29 Although appropriate treatment for infections spp. (n = 20) confirmed with aspergillosis was associated
with Candida, dimorphic fungi, or dermatophytes, it is with complete clinical resolution in 70% of the birds with
not effective against Aspergillus species.1,3,25,26,28 In African limited clinical signs, but confirmed hyphal presence, using
grey parrots, two doses (10 mg/kg and 20 mg/kg PO) were VRC orally (12.5 mg/kg BID × 44–100 days) and NE
reported from both single doses and multiple every other (20 mg/mL of saline SID × 60 minutes), and occasional
day dosing from a variety of commercial and compounded intraoperative lavage.4 However, NE VRC (10 mg/mL IV
formulation without apparent changes in FLU disposition.29 solution × 15 minutes) in pigeons resulted in no measurable
It is reportedly toxic to budgerigars (Melopsittacus undulatus) plasma concentrations.37
at doses administered safely to other psittacines.26,30 Respiratory tissues have been evaluated during nonsur-
vival VRC treatment studies and often found lacking in
Voriconazole the in vivo efficacy or direct measurement within target
A synthetic derivative of FLU, VRC is a second-generation, tissues. In racing pigeons experimentally inoculated with
broad-spectrum antifungal available in both IV and oral Aspergillus and then VRC initiated at the onset of clinical
formulations.1,17,21,26,28,31 It has additional demethylation signs (10 mg/kg BID or 20 mg/kg PO SID × 14 days),
activity that broadens its spectrum of activity.26 High VRC both doses reduced clinical signs and pathology, although
bioavailability following oral dosing occurs, and it distrib- the lower dose eliminated the Aspergillus while the higher
utes widely, including to the central nervous system.17,26 dose only reduced isolation.15 Following intratracheal
Unlike humans, who present a 30%–60% reduction in inoculation of Aspergillus, domestic quail were treated
bioavailability with food intake, plasma concentrations successfully by two doses (20 or 40 mg/kg PO SID 5–10
achieved in birds do not seem uniformly affected by feeding days). The higher dose had prolonged survival and fewer
or fasting, but effects are observed on time to reach those colony-forming units (CFUs) than control birds, but it
concentrations and duration that therapeutic concentra- was the lower dose that had significantly fewer pulmonary
tions are maintained.1,6,14,21,22 Falco spp. fed at the time of fungi.35 However, in treated mallard (Anas platyrhynchos)
VRC dosing presented 20% reduction in maximum plasma (20 mg/kg PO SID × 21 days), lung, liver, and kidney
concentrations as compared with fasted individuals.1,22 concentrations were at or less than detection concentration
African penguins (Spheniscus demersus) presented variability and no increase occurred with treatment progression.14
with food intake with a graduated elongation of elimination Neurologic dysfunction, including seizures and visual
half-life.16 Chickens had exceptionally poor VRC bioavail- disturbances, and death have been observed in six penguin
ability (<20%) and did not achieve clinically relevant species for individuals that presented plasma concentrations
plasma concentrations.1,33 Marked interspecific differences greater than 30 µg/mL that were achieved with published
in the maximum plasma concentrations achieved from the recommendations for this taxon.1,31,33 Gross and histo-
same dose administered are noteworthy and unpredictable; pathologic hepatic changes including oval cell proliferation,
for example, following 10 mg/kg PO, African grey parrots but not fibrosis, were evident in racing pigeons, includ-
achieved 1.6 µg/mL and pigeons 4.4 µg/mL at 90 minutes.6 ing clinical pathologic impact of hepatic function noted
Furthermore, despite generally shorter elimination half-lives at higher doses.31,34 Longer-term treatment in falcons has
444 SE C T I O N 12 Avian
demonstrated food flicking, anorexia, weakness, polyuria, nystatin given orally is poorly absorbed, excreted entirely
and potential effect on visual acuity sufficient to warrant unchanged in feces, and therefore essentially nontoxic at
flight restrictions during treatment.1,4,31 Polyuria also was doses of 200,000–300,000 IU/kg BID to TID × 7–10 days
observed consistently in African grey18 and Hispaniolan with recommendation of fasting before administration.5,25,26
Amazon19 (Amazon ventralis) parrots. Azole recommendations include ITRA (5–10 mg/kg PO
BID × 7–21 days); 2% miconazole gel applied directly; or
Allylamines considered most effective, FLU at 2–5 mg/kg PO SID × 7
Terbinafine days5 or 5–10 mg/kg SID × 6 weeks.26 As a flock treatment,
Represented by TERB, this class of antifungals inhibits cockatiels (Nymphicus hollandicus) were provided FLU as
ergosterol synthesis by blocking squalene monooxygen- crushed commercial tablets at 100 mg/L of drinking water
ase, causing intracellular accumulation of toxic squa- and shown by measured plasma concentrations to exceed
lene.1,3,7,25,26,28,36 It is available as topical cream, liquid spray, the MIC of 90% of Candida species with comparable clini-
or oral formulations.36 Good oral bioavailability is reported cal results with individual oral dosing as 5 mg/kg SID or
that is unaffected by feeding, although a biphasic absorption 10 mg/kg every other day (EOD).30
has been noted due to particle size during tablet dissolution, Microsporum spp. and Trichophyton spp. infections are
and TERB’s highly lipophilic and keratinophilic nature.3,24 seldom reported in birds. Typically, dermatophytosis is
Its distribution at higher doses can become limited due to managed by removal of infected keratin debris and using
saturation of tissue, although clearance was not affected.3 Its standard mammalian topical treatments.38 The earliest anti-
metabolism is not mediated by hepatic cytochrome P-450 fungal agent, griseofulvin (GF), acts by microtubule inhibi-
metabolism, but it is metabolized rapidly in the liver fol- tion, thus preventing mitosis, and has action limited solely
lowed by predominantly renal excretion, so it is subject to dermatophytes. Following oral dosing, which is enhanced
to first pass effects.1,7,25,26,28 In addition, it does present a by a fatty meal, or reduced pharmaceutical particle size, GF
biphasic elimination half-life in some avian species, where concentrates in the stratum corneum and is effective.7,25
the initial phase is less than a day but terminally extends Published avian antifungal doses for dermatophytosis
greater than 5 days.3,36 Primarily used for dermatophytosis include systemic azoles (ITRA 10 mg/kg SID × 20 days)
due to rapid accumulation within keratinocytes, other or topical polyenes and azoles.38 Topical azoles, including
fungal organisms may be susceptible, and resistance is miconazole, clotrimazole, and eniconazole, are minimally
quite rare.3,25,36 However, African grey parrots receiving oral absorbed systemically following topical absorption.1,7,25
TERB (15 and 30 mg/kg) did not achieve therapeutic drug Clotrimazole as lavage or by commercial spray or ointment
concentrations against Aspergillus species.1,36 In addition, has been published as effective.1,26 Eniconazole is produced
documented TERB NE (1 mg/mL solution for 15 minutes) in a premise treatment formulation that should not be used
in Hispaniolan Amazon parrots using suspensions made extralabel as a pharmaceutic, although compounded topical
from either crushed tablet or raw drug powder persisted products are available outside the United States.7,26
above the Aspergillus sp. MIC only for 1 hour or 4 hours,
respectively,37 due to the known poor dissolution of TERB Macrorhabdus Ornithogaster39
and its settling in suspension.1,36
This anamorphic ascomycetal yeast grows exclusively at the
Flucytosine7,25,26 proventricular-ventricular junction and produces maldiges-
In fungi that contain cytosine permease, this pharmaceutic tion and weight loss. In reported treatments the measure
is converted cytoplasmically to 5-fluorouracil and inhibits of success was cessation of fecal shedding of organism. By
RNA synthesis. Hepatotoxicity or bone marrow suppression gavage, AmpB (25 mg/kg PO BID × 14 days) has been used
can occur in the patient as a result of this conversion within and appears safe and rapidly effective when compounded
the gastrointestinal tract. Orally, it is effective against yeast from commercially available powder, although some failure
only, although it is well-absorbed and distributed, includ- has been reported at much higher doses (100–150 mg/kg
ing extensively to the CSF. However, it rapidly develops PO BID × 30 days).26 Nystatin also has been used as a flock
resistance and must be used only in combination with other treatment at 3,500,000 IU/L of drinking water × 2 days
antifungal agents. and then for another 28 days at 2,000,000 IU/L. Although
FLU at 100 mg/kg PO was successful experimentally in
treatment of chickens, this dose was toxic in budgerigars
Non-Aspergillus Fungi Treatment and not effective at lower doses in this species.
Mucosal and Dermal Fungal Infections
The most common fungal infections of mucosal surfaces References
of the cranial gastrointestinal tract and nasal sinus result
from Candida species. Treatment is most successful when 1. Krautwald-Junghanns M-E, Vorbrüggen S, Böhme J: Aspergil-
the antifungal agent has direct contact with the fungus.5 losis in birds: an overview of treatment options and regimens, J
Although quite toxic parenterally, the polyene antifungal Exot Pet Med 24:296–307, 2015.
CHAPTER 62 Antifungals in Birds 445
2. Redig P: Aspergillosis. In Samour J, editor: Avian medicine, ed 20. Gentry J, Montgerard C, Crandall E, et al: Voriconazole disposi-
3, St. Louis, 2016, Elsevier, pp 460–472. tion after single and multiple, oral doses in healthy, adult red-
3. Bechert U, Christiansen JM, Poppenga R, et al: Pharmacoki- tailed hawks (Buteo jamaicensis), J Avian Med Surg 28:201–208,
netics of orally administered terbinafine in African penguins 2014.
(Spheniscus demersus) for potential treatment of aspergillosis, J 21. Parsley RA, Tell LA, Gehring R: Pharmacokinetics of a single oral
Zoo Wildl Med 41:263–274, 2010. dose of voriconazole administered orally with and without food
4. DiSomma A, Bailey T, Silvanose C, et al: The use of voriconazole to red-tailed hawks (Buteo jamaicensis), Am J Vet Res 78:433–439,
for the treatment of aspergillosis in falcons (Falco species), J 2016.
Avian Med Surg 21:307–316, 2007. 22. Schmidt V, Demiraj F, Di Somma A, et al: Plasma concentration
5. Silvanose C: Candidiasis. In Samour J, editor: Avian medicine, of voriconzole in falcons, Vet Rec 161:265–268, 2007.
ed 3, St. Louis, 2016, Elsevier, pp 472–473. 23. Evans EE, Emery LC, Cox SK, et al: Pharmacokinetics of terbi-
6. Burhenne J, Scope A: Evaluation of the influence of formulation, nafine after oral administration of a single dose to Hispaniolan
food intake and species on voriconazole plasma concentration in Amazon parrots (Amazona ventralis), Am J Vet Res 74:835–838,
birds, Vet Med Austria 99:157–162, 2012. 2013.
7. Duncan M: Fungal diseases in all taxa. In Fowler ME, Miller 24. Bechert U, Christiansen JM, Poppenga R, et al: Pharmacokinet-
RE, editors: Zoo and wild animal medicine, ed 5, St. Louis, 2003, ics of terbinafine after single oral dose administration in red-
Elsevier, pp 727–731. tailed hawks (Buteo jamaicensis), J Avian Med Surg 24:122–130,
8. Tell LA, Craigmill AL, Clemons KV, et al: Studies on itracon- 2010.
azole delivery and pharmacokinetic in mallard ducks (Anas 25. Bossche HV, Engelen M, Rochette F: Antifungal agents of use
platyrhynchos), J Vet Pharmacol Ther 28:267–274, 2005. in animal health – chemical, biochemical and pharmacological
9. Lumeij JT, Gorgevska D, Woestenborghs R: Plasma and tissue aspects, J Vet Pharmacol Ther 26:5–29, 2003.
concentrations of itraconazole in racing pigeons (Columba livia 26. Plumb DC: Plumb’s veterinary drug handbook, ed 8, Hoboken,
domestica), J Avian Med Surg 9:32–35, 1995. NJ, 2015, Wiley-Blackwell.
10. Smith JA, Papich MG, Russell G, et al: Effects of compounding 27. Burhenne J, Haefeli WE, Hess M, et al: Pharmacokinetics, tissue
on pharmacokinetics of itraconazole in black-footed penguins concentrations, and safety of the antifungal agent voriconazole in
(Spheniscus demersus), J Zoo Wildl Med 41:487–495, 2010. chickens, J Avian Med Surg 22:199–207, 2008.
11. Bunting EM, Madi NA, Cox S, et al: Evaluation of oral itracon- 28. Keller KA: Therapeutic review: itraconazole, J Exot Pet Med
azole administration in captive Humboldt penguins (Spheniscus 20:156–160, 2011.
humboldti), J Zoo Wildl Med 40:508–519, 2009. 29. Guzman DS-M: Advances in avian clinical therapeutics, J Exot
12. Orosz SE, Frazier DL, Schroeder EC, et al: Pharmacokinetic Pet Med 23:6–20, 2014.
properties of itraconazole in blue-fronted Amazon parrots 30. Flammer K, Papich M: Pharmacokinetics of fluconazole after
(Amazona aestiva aestiva), J Avian Med Surg 10:168–173, 1996. oral administration of single and multiple doses in African grey
13. Jones MP, Orosz SE, Cox SK, et al: Pharmacokinetic disposition parrots, Am J Vet Res 67:417–422, 2006.
of itraconazole in red-tailed hawks (Buteo jamaicensis), J Avian 31. Ratzlaff K, Papich MG, Flammer K: Plasma concentrations of
Med Surg 14:15–22, 2000. fluconazole after a single oral dose and administration in drink-
14. Kline Y, Clemons KV, Woods L, et al: Pharmacokinetics of ing water in cockatiels (Nymphicus hollandicus), J Avian Med Surg
voriconazole in adult mallard ducks (Anas platyrhynchos), Med 25:23–31, 2011.
Mycol 49:500–512, 2011. 32. Sim RR: Voriconazole, J Exot Pet Med 25:342–347, 2016.
15. Beernaert LA, Pasmans F, Baert K, et al: Designing a treatment 33. Hyatt MW, Georoff TA, Nollens HH, et al: Voriconazole toxic-
protocol with voriconazole to eliminate Aspergillus fumigatus from ity in multiple penguin species, J Zoo Wildl Med 46:880–888,
experimentally inoculated pigeons, Vet Microbiol 139:393–397, 2015.
2009. 34. Beernaert LA, Baer K, Marin P, et al: Designing voriconazole
16. Hyatt MW, Wiederhold NP, Hope WW, et al: Pharmacokinetics treatment for racing pigeons: balancing between hepatic enzyme
of orally administered voriconazole in African penguins (Sphe- auto induction and toxicity, Med Mycol 47:276–285, 2009.
niscus demersus) after single and multiple doses, J Zoo Wildl Med 35. Tell LA, Clemons KV, Kline Y, et al: Efficacy of voriconazole in
48(2):352–362, 2017. Japanese quail (Coturnix japonica) experimentally infected with
17. Parsley RA, Mutlow AG, Tell LA, et al: Comparison of different Aspergillus fumigatus, Med Mycol 48:234–244, 2010.
pharmacokinetic models’ predictions of plasma voriconazole 36. Keller KA: Therapeutic review: terbinafine, J Exot Pet Med
concentrations in a Magellanic penguin following long-term oral 21:181–185, 2012.
administration; 2017; in preparation. 37. Emery LC, Cox SK, Souza MJ: Pharmacokinetics of nebulized
18. Flammer K, Nettifee Osborne JA, Webb DJ, et al: Pharmaco- terbinafine in Hispaniolan Amazon parrots (Amazona ventralis),
kinetics of voriconazole after oral administration of single and J Avian Med Surg 26:161–166, 2012.
multiple doses in African grey parrots (Psittacus erithacus timneh), 38. Samour J: Favus or ringworm infection. In Samour J, editor:
Am J Vet Res 69:114–121, 2008. Avian medicine, ed 3, St. Louis, 2016, Elsevier, pp 477–479.
19. Guzman DS-M, Flammer K, Papich MG, et al: Pharmacokinetics 39. Phalen D: Macrorhabdus ornithogaster. In Samour J, editor:
of voriconazole after oral administration of single and multiple Avian medicine, ed 3, St. Louis, 2016, Elsevier, pp 473–477.
doses in Hispaniolan Amazon parrots (Amazon ventralis), Am J
Vet Res 71:460–467, 2010.
63
Medical Management of
Walk-Through Aviaries
MEREDITH MARTIN CLANCY
W
alk-through aviaries are an expanding part of colored Australian birds are found throughout the United
the interactive experiences seen at zoological States and around the world.6–7 Budgerigars are smaller
institutions around the world. Perhaps the first with a different feeding strategy than lorikeets (Trichoglossus
walk-through aviary was built by Smithsonian Institute to spp.). Budgerigars are granivorous,8 with interactive aviaries
house birds at the St. Louis World’s Fair in 1904. Thought typically offering seed sticks that are prepared in-house.6–7
to be the largest walk-through aviary at the time, it was With a Latin name that describes their brushy tongue spe-
purchased by the Saint Louis Zoo, where it currently cially designed to feed on nectar and pollen, lorikeets are
remains.1 Now, walk-through aviaries are common and fed cups of commercially prepared nectar by guests at many
represent a shift in focus toward guest-animal interaction institutions. Other walk-through aviaries may house other
where, through an emphasis on education, guest engage- small-to-medium-sized psittacines like cockatiels (Nym-
ment and entertainment may drive conservation missions phicus hollandicus) in guest interactive displays similar to
at the heart of zoos’ higher purpose.2–4 This chapter will those described for budgies.6 Wider variety of bird species,
cover the medical management of walk-through aviaries, such as passerines like Australian finches (Estrildidae) and
especially addressing the complexities added in those aviar- weavers (Ploceidae), columbiformes, and ground birds like
ies with an interactive guest component. Walk-through galliformes and anseriformes, are found in walk-through
aviary medical management combines preventive care of aviaries without structured guest feeding. Although human
the flock and interventional medicine of the individual interaction is possible in these aviaries, the focus is primarily
bird with public health best practices. A successful walk- on safe interactions between guests and birds and providing
through aviary starts with appropriate exhibit design for species-appropriate design for the exhibit.8–11
the level of interaction desired, safe animal sourcing and
quarantine, and an evidence-based flock surveillance pro- Exhibit Design
tocol with adequate medical intervention capabilities on
an individual and population level. It must also include Veterinary involvement in the design aspects of a walk-
planning with the appropriate public health entities for through aviary may mitigate future medical and husbandry
guest education, zoonotic disease prevention, and disease concerns. Examples include recommendations that reduce
outbreak management. pathogen persistence and transmission and the development
of husbandry protocols to refine and improve health and
Species Choice nutrition. Adequate provision for space and understanding
the species’ natural history may reduce competition over
Species choice is an important step in designing a suc- nesting sites, feeding stations, protection from inclement
cessful exhibit. Zoo standards and accreditation guidelines weather, and other resources and allow for desired social
may direct aviary exhibit design and population choices. behavior, including with guests. During aviary operation,
Although guests must always be supervised in animal contact Association of Zoos and Aquariums (AZA) and European
settings, even with the best observation, certain bird orders, Association of Zoos and Aquaria standards require zoo staff
such as Falconiformes, Accipitriformes, and Strigiformes supervision.4–5 This helps to safeguard both guests and birds
are inappropriate for aviaries where guests share space with from preventable trauma and inappropriate interaction.
the birds.4–5 Species of birds managed in interactive aviaries Traumatic injuries from conspecific aggression, exhibit
are primarily small psittacines, generally of the subfamily structures, or, sadly, inadvertent guests, should be cataloged
Loriinae: budgerigars (Melopsittacus undulatus) and lories so that keepers, exhibit operations, and other stakehold-
and lorikeets (tribe Loriini). Exhibits with these brightly ers may mitigate problems. Given the predilection for
446
CHAPTER 63 Medical Management of Walk-Through Aviaries 447
TABLE
63.1 Avian Pathogens and Their Appropriate Disinfectants
Quaternary
Ammonium Phenols/
Disease (Etiologic Agent) Oxidizing Agents Compounds Aldehydes Phenolics Alcohols
Psittacine beak and feather Chlorine, Sodium Glutaraldehyde8,44 Effective77
disease (BFDV, Circoviridae) hypochlorite,8 e.g.,
Virkon S31
Exotic Newcastle disease Chloramine 2% Formalin8 Effective,77 e.g.,
(APMV-1, Paramyxoviridae) 1%, Sodium 1% Lysol8
hypochlorite8
Avian influenza Effective77 Effective8,10 Effective77 Effective77
(AIV, Orthomyxoviridae)
Avian polyomavirus (APV, Sodium Effective77 Synthetic 70%
Polyomaviridae) hypochlorite,8,44 phenol,44 ethanol44
stabilized chlorine phenolics8,77
dioxine44
Avian mycobacteriosis Formaldehyde50,77 Effective,9 e.g.,
(Mycobacterium avium Sodium-o-
complex, Firmicutes) phenylphenol50
G+ve bacteria Clorox9 Effective,50 e.g., Effective77 Effective,77e.g., Effective77
(e.g., Erysipelothrix, Roccal9 sodium-o-
Clostridium, Staphylococcus phenylphenol,50
spp., Firmicutes) One-stroke9
Bacterial Spores Variable77 Effective77 Effective77
(e.g., Clostridium spp.)
G-ve bacteria Sodium hypochlorite8 Limited,75 Effective77 Effective,77 e.g.,
(e.g., Enterobacteriaceae, Clorox9 including One-stroke9
Pasteurella, Proteobacteria) Roccal9
Chlamydiosis/Psittacosis 3% hydrogen Effective,8 e.g., 1% Lysol8 70%
(Chlamydia psittaci, peroxide,8 1:32 benzalkonium ethanol8
Chlamydiae) dilution of sodium chloride28
hypochlorite49
Cryptosporidiosis Oxine* Chloro-m-cresol
(Cryptosporidium spp., (more than
Apicomplexa) bleach)39
Flock Nutrition and Husbandry Another mechanism of maintaining bird health is through
maintaining a balance in the normal gastrointestinal flora.
Feeder design is equally as important as what goes in the Avian probiotics are gaining favor with some practitioners
feeder because nutrition is an important factor in avian as a means of managing proliferation of a single organism
health.12,34 Food items offered by guests may represent only over traditional medical management with broad-spectrum
one aspect of the birds’ diet or may be the complete diet. antibiotics.34,37–39 Although product quality and labeling
The interactive portion allows for increased variability in accuracy have been called into question, probiotic use has
amount and, if guest offerings comprise only one aspect of been increasing in avian medicine.40 An outbreak of attach-
the diet, proportion of the balanced diet.5 Birds in walk- ing and effacing Escherichia coli in an interactive budgerigar
through aviaries may become obese or develop mineral aviary was managed through use of probiotic and exhibit
imbalances through selection of high-energy seeds or and husbandry modifications. The goal of this management
through consumption of high-concentration nectar from strategy was not to eliminate the pathogen in the aviary, but
guests 6,8,35–36 Provisions for complete diet and a viable means to reduce shedding and morbidity in the flock (Box 63.3).15
to monitor individual body condition and nutritional status Continuous reassessment and refinement of aviary pro-
must be made by the veterinarian, in conjunction with the tocols may uncover problems and help to navigate solutions.
nutritionist and animal care managers. An increase in morbidity and mortality in an interactive
CHAPTER 63 Medical Management of Walk-Through Aviaries 449
• BOX 63.3 Escherichia coli Sepsis/ disease screening should be a priority. AZA accreditation
Gastrointestinal Disease guidelines specifically highlight chlamydiosis in birds of
the orders Psittaciformes, Galliformes, and Columbiformes
Zoonotic and salmonellosis in all avian species as the predominant
Etiology: Escherichia coli concerns for guest contact zoonosis risk.4
Susceptible: All species
Clinical signs: Acute death, weight loss, diarrhea
Pathophysiology: Enteritis, hepatitis, sepsis Screening for Zoonotic Agents
Diagnostics: Enteric culture gold standard
Treatment: Reduce shedding via antibiotics Chlamydiosis shedding via nasal secretions and feces is
Prevention: Probiotics, reduce fecal-oral transmission exacerbated by stress, including shipping and crowding,
making quarantine a high-risk period.46 Although screen-
ing tests remain imperfect, the standard serologic test for
chlamydial antibodies is reported to be the modified direct
budgerigar aviary was initially ascribed to trauma. Thor- complement fixation (CF), with a fourfold increase in
ough review of nutrition and husbandry and evaluation of paired titer samples considered diagnostic and clinical signs,
individuals antemortem and postmortem identified the true such as upper respiratory disease, with a single high titer
cause as feed-related hypervitaminosis D. Manufacturer adequate for presumptive diagnosis.28,46–48 Budgerigars and
error in vitamin D3 supplementation resulted in soft tissue cockatiels are among the most commonly reported avian
mineralization causing nonspecific clinical signs. Persistence species testing positive for chlamydiosis.28 In lieu of flock
was required to identify the true cause of mortalities in the screening, some institutions will routinely treat high-risk
flock. This underscores the importance of close scrutiny or public-contact birds.28,41,47,49–51 Traditionally, treatment
of exhibits and husbandry practices, with added value to has consisted of a tetracycline for 45 days, a duration
written protocols that may be routinely evaluated.41 that exceeds two complete avian macrophage replication
Walk-through aviaries with guest interaction have cycles, allowing for chlamydial organisms to be released
a relatively high density of animals in the aviary space, from the macrophage during division where—at adequate
allowing for more reliable and repeatable guest experiences.6 plasma concentrations—inhibition of C. psittaci occurs.28,47
This flock density increases the ability to transmit patho- Although the risk of zoonotic spread of chlamydiosis from
gens and the rapidity with which pathogens may spread. a walk-through aviary is considered low, institutions should
Understanding the flock’s population health and following invite input from appropriate public health officials prior
ideal avicultural principles through appropriate quarantine to instituting a screening or treatment protocol.47 However,
and animal sourcing are paramount to reducing risk and any bird with clinical signs should be isolated from the flock
establishing a successful aviary.9–10,42 and guests and screened to reduce risks both medical and
legal (Box 63.4).
Animal Sourcing and Quarantine Zoonoses such as salmonellosis are generally screened
for via fecal testing.29–30,52 Given the risk presented to the
Zoos often source flocks for walk-through aviaries from public and keepers, screening tests with high sensitivity are
private breeders or commercial entities.7,41,43 If a private or preferential. Group or pooled samples decrease cost and
commercial source is to be used to stock a zoo’s aviary, one may increase likelihood of identifying the disease at the
that adheres to the Model Aviary Program (MAP) would population level to counteract the reduced sensitivity intro-
be ideal to establish a baseline for medical and husbandry duced by intermittent individual shedding.9,53 At SDZSP,
practices.7,10,42 The MAP was developed to educate aviary Salmonella fecal PCR is used to screen for salmonellosis
owners and veterinarians about reducing risk of chlamydio- in lorikeets both as a routine and medical diagnostic test.
sis spread, but it may be used to mitigate risks from any Sensitivity is increased by collecting daily samples for 3
infectious disease. It calls for maintenance of accurate days. Although the molecular testing may result in a false
records of aviary transactions and avoidance of mixing positive due to pass-through of a nonviable organism, the
birds from multiple sources, along with isolation and benefit of increased sensitivity and quicker turnaround time
testing of any bird with abnormal clinical presentation.42 If as compared with fecal culture helps to guide more rapid
medical history of the institution or breeder or preshipment intervention to reduce risk to other birds and guests (Box
testing is inadequate or unavailable, the quarantine period 63.5).54
should be extended beyond the incubation period of avian
pathogens of concern or allow for sufficient screening of a
representative segment of the flock.29,42–43 Because some of Additional Disease Surveillance and
the species are too small to allow for serology or even routine Medical Intervention
complete blood count and biochemistry panels, quarantine
screening should be risk-based and target diseases specific to Surveillance of the flock should be evidence-based, guided by
the species or institution’s collection.9,43–45 Given the guest species- and institution-specific health concerns.9–10 Because
interaction component of the aviaries in focus here, zoonotic whole flock testing may prove logistically challenging, often
450 SE C T I O N 12 Avian
the environment because these are not obligate ectoparasites • BOX 63.7 Reportable Avian Viruses
and can include nonpharmaceutical alternatives, such as
diatomaceous earth, predatory mites, and botanical deter- Avian Influenza
rents (Lotz and Zuba, personal communication). Zoonotic
Etiology: Avian influenza virus (AIV), type A influenza virus,
Orthomyxoviridae
Public Health Susceptible: All, Charadriiformes and Anseriformes considered
reservoirs, Galliformes more susceptible
This chapter provides insight into some of the pathogens Clinical Signs: Low-path avian influenza (LPAI)—respiratory
and noninfectious diseases that may affect birds in walk- disease; high-path AI (HPAI)—acute death, systemic illness
Pathophysiology: Respiratory or gastrointestinal disease; HPAI
through aviaries, but the list is not exhaustive. Each aviary
systemic
will need an individualized approach developed through Diagnostics: Reverse-transcriptase PCR (RT-PCR) matrix (M)
collaboration between the veterinarian and the husbandry protein
team. The attending veterinarian should also coordinate
with state and local public health officials regarding the Exotic Newcastle Disease
aviary’s preventive medicine practices and flock surveillance. Zoonotic
Etiology: Avian paramyxovirus 1 (APMV-1), virulent Newcastle
A systematic process for managing zoonotic disease
disease virus, Paramyxoviridae
concerns should be designed and documented for each Susceptible: All, Suliformes, Psittaciformes, Columbiformes,
institution with contact animals.50 The National Associa- Galliformes most susceptible
tion of State Public Health Veterinarians (NASPHV) has Clinical signs: Respiratory disease, nervous signs, diarrhea,
developed signage and education materials that may be depression, high mortality
Pathophysiology: Respiratory, gastrointestinal, neurologic,
used for many animal contact exhibits.47 Well-established
reproductive disease; lymphoid necrosis
Centers for Disease Control and Prevention (CDC) and Diagnostics: Viral isolation, hemagglutination inhibition, RT-PCR
NASPHV recommendations are defensible standards of Treatment: Supportive care, including antibiotics to treat
care when dealing with the legal implications of zoonoses secondary bacterial pathogens
in contact animal exhibits.70 Apart from federally report- Prevention: Reduce contact with reservoir species to stop
transmission; vaccination may be available to control
able diseases, laws differ from one jurisdiction to the other
outbreak
on what constitutes a reportable disease or animal injury/
bite. Most zoos have an onsite human health official or
a contracted health service, and coordination with these
individuals, as well as local public health officials, may keep
guidelines up to date. flock and fostering a thriving guest experience cannot be
Hand-washing is an essential first line of defense for underestimated.
guests in an interactive aviary.29,47 Even with adequate
signage and staff monitoring, adherence to preventive prac- References
tices by guests may be variable.71–72 Although from a public
health official perspective these guidelines are established to 1. Leonard MD: Animals always: 100 years at the Saint Louis Zoo.
protect the guests from the bird, the attending veterinarian Columbia, MO, 2009, University of Missouri.
must consider protecting the birds from disease introduc- 2. Luebke JF, Matiasek J: An exploratory study of zoo visitors’
tion from guests. Avian influenza and exotic Newcastle exhibit experiences and reactions, Zoo Bio 32:407–416, 2013.
3. Povey K, Rios J: Using interpretive animals to deliver affective
disease (END) are viral agents that may be introduced to
messages in zoos, J Interpret Res 7:19–28, 2002.
a flock inadvertently (Box 63.7).73 Significantly heightened 4. Association of Zoos & Aquariums: The accreditation standards
biosecurity was implemented at SDZSP during a local and related policies, 2017 edition. Available at: www.aza.org/
END outbreak.74 Through the Zoo and Aquarium All assets/2332/aza-accreditation-standards.pdf. (Accessed 18 March
Hazards Preparedness, Response, and Recovery (ZAHP) 2017).
Fusion Center, avian influenza zoo preparedness exercises 5. European Association of Zoos and Aquaria: Standards for the
are available, along with contingency planning for more Accommodation and Care of Animals in Zoos and Aquaria,
than just disease outbreaks.75–76 approved 27 September 14. Available at: www.eaza.net/
The operational value to the institution of a walk-through assets/Uploads/Standards-and-policies/Standards-for-the
aviary is an important consideration when managing a -Accommodation-and-Care-of-Animals-2014.pdf. (Accessed 14
suspected or confirmed zoonotic disease. Established and February 2017).
6. Avian Scientific Advisory Group: Lorikeet/Budgie Questionnaire.
documented triggers may make the difficult decision to
Available at: http://aviansag.org/TAG/Parrot/Lorikeet.htm.
close the aviary to guests less clouded by financial or other (Accessed 18 December 2016).
implications. The long-term success of the aviary depends 7. Hanna S: Interactive lorikeet exhibits… a behind the scenes look,
on effective management of the problems as discussed in J Am Feder Aviculture 34:38–43, 2007.
this chapter through public health vigilance, flock surveil- 8. Heatley JJ, Cornejo J: Psittaciformes. In Fowler ME, Miller RE,
lance and medical management, and appropriate aviary editors: Fowler’s zoo and wild animal medicine (vol 8). St. Louis,
husbandry. The veterinarian’s role in maintaining a healthy 2015, Elsevier, pp 172–186.
452 SE C T I O N 12 Avian
9. Clubb SL, Flammer K: The avian flock. In Ritchie BW, 28. Crosta L, Melillo A, Schnitzer P: Chlamydiosis (Psittacosis). In
Harrison GJ, Harrison LR, editors: Avian medicine: principles Speer BL, editor: Current therapy in avian medicine and surgery.
and application. Lake Worth, FL, 1994, Wingers Publishing, St. Louis, 2016, Elsevier, pp 82–94.
pp 46–61. 29. James SB: Children’s zoo medicine: zoonoses. In Miller RE,
10. Phalen DN: Preventive medicine and screening. In Harrison GJ, Fowler ME, editors: Fowler’s zoo and wild animal medicine current
Lightfoot TI, editors: Clinical avian medicine. Palm Beach, FL, therapy (vol 7). St. Louis, 2012, Elsevier, pp 115–124.
2006, Spix, pp 573–586. 30. Kirkwood JK: Salmonellosis in songbirds (order Passeriformes).
11. Smith JA: Passeriformes (Songbirds, Perching Birds). In Fowler In Fowler ME, Miller RE, editors: Zoo and wild animal medicine
ME, Miller RE, editors: Fowler’s zoo and wild animal medicine (vol 6). St. Louis, 2008, Elsevier, pp 166–169.
(vol 8). St. Louis, 2015, Elsevier, pp 236–246. 31. Raidal S: Psittacine beak and feather disease. In Speer BL, editor:
12. Koutsos E: Foundations in avian nutrition. In Speer BL, editor: Current therapy in avian medicine and surgery. St. Louis, 2016,
Current therapy in avian medicine and surgery. St. Louis, 2016, Elsevier, pp 51–59.
Elsevier, pp 142–150. 32. Shivaprasad HL, Palmieri C: Pathology of mycobacteriosis in
13. Langan J: Integrated pest management. In Miller RE, Fowler birds, Vet Clin North Am Exot Anim 15:41–55, 2012.
ME, editors: Fowler’s zoo and wild animal medicine current therapy 33. Gelis S: Advancements in nutrition of Loridae. In Speer BL,
(vol 7). St. Louis, 2012, Elsevier, pp 51–59. editor: Current therapy in avian medicine and surgery. St. Louis,
14. Riggs G: Avian mycobacterial disease. In Miller RE, Fowler ME, 2016, Elsevier, pp 150–164.
editors: Fowler’s zoo and wild animal medicine current therapy (vol 34. Nemeth NM, Gonzalez-Astudillo V, Oesterle PT, et al: A 5-year
7). St. Louis, 2012, Elsevier, pp 66–274. retrospective review of avian diseases diagnosed at the Depart-
15. Seeley KE, Baitchman E, Bartlett S, et al: Investigation and ment of Pathology, University of Georgia, J Comp Path 1:1–16,
control of an attaching and effacing Escherichia coli outbreak in 2016.
a colony of captive budgerigars (Melopsittacus undulatus), J Zoo 35. Gyimesi Z, Garner MM, Burns RB, et al: Hepatopathy in
Wildl Med 45:875–882, 2014. zoo Lorinae: is diet-induced hypervitaminosis A the cause? In
16. Travis D: West Nile virus in birds and mammals. In Fowler ME, Proceedings of the 42nd Annual American Association of Zoo
Miller RE, editors: Zoo and wild animal medicine current therapy Veterinarians/American Association of Wildlife Veterinarians
(vol 6). St. Louis, 2008, Elsevier, pp 2–9. Conference, 2019, p 215.
17. deMaar TW, Snowden KF: The avian sarcocystis predator: not 36. Kalmar ID, van Loon M, Bürkle M, et al: Effect of dilution
the species we think. In Proceedings of the 44th Annual Confer- degree of commercial nectar and provision of fruit on food,
ence of the American Association of Zoo Veterinarians, 2012, pp energy and nutrient intake in two rainbow lorikeet subspecies,
195–196. Zoo Biol 28:98–106, 2009.
18. Persky M, Suedmeyer WK: Outbreak of sarcocystosis in rainbow 37. Doneley RJT: Bacterial and parasitic diseases of parrots, Vet Clin
lorikeets (Trichoglossus heamatodus [sic]) housed in an outdoor Exot Anim 12:417–432, 2009.
aviary. In Proceedings of the 43rd Annual Conference of the 38. Phalen DN: Update: diagnosis and management of Macrorhab-
American Association of Zoo Veterinarians, 2011, p 94. dus ornithogaster (formally Megabacteria), Vet Clin North Am Exot
19. Rimoldi G, Speer B, Wellehan JFX, et al: An outbreak of Sarco- Anim 17:203–210, 2013.
cystis calchasi encephalitis in multiple psittacine species within an 39. Wellehan JFX: The pathogenesis of infectious diseases. In Speer
enclosed zoological aviary, J Vet Diagn Invest 25:775–781, 2013. BL, editor: Current therapy in avian medicine and surgery. St.
20. Siegal-Willot JL, Pollock CG, Carpenter JW, et al: Encephalitis Louis, 2016, Elsevier, pp 22–27.
caused by Sarcocystis falcatula-like organisms in a white cockatoo 40. Echols MS: Navigating the nutraceutical industry: a guide to
(Cacatua alba), J Avian Med Surg 1:19–24, 2005. help veterinarians make informed clinical decisions. In Speer BL,
21. Villar D, Kramer M, Howard L, et al: Clinical presentation and editor: Current therapy in avian medicine and surgery. St. Louis,
pathology of sarcocystosis in psittacine birds: 11 cases, Avian Dis 2016, Elsevier, pp 164–176.
52:187–194, 2008. 41. Olds JE, Burrough E, Madson D, et al: Clinical investigation into
22. Cray C, Zielezienski-Roberts K, Bonda M, et al: Serologic feed-related hypervitaminosis D in a captive flock of budgerigars
diagnosis of sarcocystosis in psittacine birds: 16 cases, J Avian (Melopsittacus undulatus): morbidity, mortalities, and pathologic
Med Surg 19:208–215, 2005. lesions, J Zoo Wildl Med 46:9–17, 2015.
23. Dykstra MJ, Reininger K: Aviary air-handler design and its 42. Model Aviculture Program: Available at: modelaviculture.org.
relationship to fungal spore loads in the air, J Zoo Wildl Med (Accessed 17 March 2017).
38:540–547, 2007. 43. Perpiñán D, Garner MM, Wellehan JFX, et al: Mixed infec-
24. National Association of State Public Health Veterinarians Animal tion with reovirus and Chlamydophila in a flock of budgerigars
Contact Compendium Committee: Compendium of measures (Melopsittacus undulatus), J Avian Med Surg 24:316–321, 2010.
to prevent disease associated with animals in public settings, 44. Raidal SR: Avian circovirus and polyomavirus diseases. In Miller
2013, J Am Vet Med Assoc 243:1270–1288, 2013. RE, Fowler ME, editors: Fowler’s zoo and wild animal medicine
25. Phalen D: Psittacid herpesvirus and associated diseases. In current therapy (vol 7). St. Louis, 2012, Elsevier, pp 297–303.
Speer BL, editor: Current therapy in avian medicine and surgery. 45. Ramsay EC, Steeil J, Cross T, et al: A risk-based quarantine
St. Louis, 2016, Elsevier, pp 47–51. program: an alternative to one size fits all. In Proceedings of
26. Vitali SD, Eden PA, Payne KL, et al: An outbreak of mycobacte- the Annual Conference of the Association of Zoo Veterinarians,
riosis in Gouldian finches caused by Mycobacterium peregrinum, 2013, p 131.
Vet Clin Exot Anim 9:519–522, 2006. 46. Wallace C, Marinkovich M, Morris PJ, et al: Lessons from a
27. Witte CL, Hugerford LL, Papendick R, et al: Investigation of retrospective analysis of a 5-yr period of quarantine at San Diego
characteristics and factors associated with avian mycobacteriosis Zoo: a risk-based approach to quarantine isolation and testing
in zoo birds, J Vet Diagn Invest 20:186–196, 2008. may benefit animal welfare, J Zoo Wildl Med 47:291–296, 2016.
CHAPTER 63 Medical Management of Walk-Through Aviaries 453
47. American Association of Zoo Veterinarians: Guidelines for zoo and 62. Pizarro M, Höfle U, Rodríguez-Bertos A, et al: Ulcerative
aquarium veterinary medical programs and veterinary hospitals, ed enteritis (quail disease) in lories, Avian Dis 49:606–608, 2005.
6, 2016. Available at: c.ymcdn.com/sites/www.aazv.org/resource/ 63. Raymond JT, Topham K, Shirota K, et al: Tyzzer’s disease in a
resmgr/files/aazvveterinaryguidelines2016.pdf. (Accessed 18 neonatal rainbow lorikeet (Trichoglossus haematodus), Vet Pathol
December 2016). 38:326–327, 2001.
48. Cray C: Infectious and zoonotic disease testing in pet birds, Clin 64. Shivaprasad HL, Cadenas MB, Diab SS, et al: Coxiella-like infec-
Lab Med: 71–85, 2011. tion in psittacines and a toucan, Avian Dis 52:426–432, 2008.
49. National Association of State Public Health Veterinarians: Com- 65. Woc-Colburn AM, Garner MM, Bradway D, et al: Fatal coxiel-
pendium of Measures to control Chlamydophila psittaci infection losis in Swainson’s blue mountain rainbow lorikeets (Trichoglossus
among humans (Psittacosis) and pet birds (avian chlamydio- haematodus moluccanus), Vet Pathol 45:247–254, 2008.
sis), 2010. Available at: http://www.nasphv.org/Documents/ 66. Marinkovich M, Wallace C, Morris PJ, et al: Lessons from a
Psittacosis.pdf. (Accessed 18 June 2016). retrospective analysis of a 5-yr period of preshipment testing at
50. Oie Manual of Diagnostic Tests and Vaccines for Terrestrial San Diego Zoo: a risk-based approach to preshipment testing
Animals 2017. Available at: http://www.oie.int/en/international may benefit animal welfare, J Zoo Wildl Med 47:297–300, 2016.
-standard-setting/terrestrial-manual/access-online/. (Accessed 18 67. Garner MM: A retrospective study of diseases in captive lories
December 2016). and lorikeets. In Proceedings of the Association of Avian Veteri-
51. Janssen DL: Guidelines for the management of zoonotic diseases. narians Conference, 2008, p 65.
In Miller RE, Fowler ME, editors: Fowler’s zoo and wild animal 68. Amin A, Bilic I, Liebhart D, et al: Trichonomads in birds—a
medicine (vol 8). St. Louis, 2015, Elsevier, pp 733–734. review, Parasitology 141:733–747, 2014.
52. Ward MP, Ramer JC, Proudfoot J, et al: Outbreak of salmonel- 69. Brandão J, Beafrère H: Clinical update and treatment of selected
losis in a zoologic collection of lorikeets and lories (Trichoglossus, infectious gastrointestinal diseases in avian species, J Exot Pet Med
Lorius, and Eos spp.), Avian Dis 47:493–498, 2003. 22:101–117, 2013.
53. Thurmond MC: Conceptual foundations for infectious disease 70. Babcock DW: Legal implications of zoonotic-disease outbreaks
surveillance, J Vet Diagn Invest 15:501–514, 2003. at petting zoos and animal exhibits, J Environ Health 46–47,
54. Zuba JR, Singleton C, Papendick R: Avian chlamydiosis 2006.
and doxycycline toxicity in a flock of free-contact rainbow 71. Marion C: Human-animal interactions at zoological institutions.
lorikeets (Trichoglossus haematodus haematodus): considerations, Kansas State University Masters of Public Health Thesis, 2011.
concerns and quandaries. In Proceedings of the 33rd Annual Available at: http://krex.k-state.edu/dspace/handle/2097/9198.
American Association of Zoo Veterinarians Conference, 2002, pp (Accessed 18 December 2016).
105–108. 72. McMillian M, Dunn JR, Keen JE, et al: Risk behaviors for
55. Clancy MM, Lamberski N: Minimizing risk by maximizing disease transmission among petting zoo attendees, J Amer Vet
intervention to prevent Salmonella in aviary rainbow lorikeets Med Assoc 231:1036–1038, 2007.
(Trichoglossus haematodus). In Proceedings of the 49th Annual 73. California Department of Food and Agriculture Animal
American Association of Zoo Veterinarians Conference, 2017, p Health Branch: Biosecurity guidelines to prevent Exotic
26. Newcastle Disease Virus. Available at: https://www.cdfa.ca.gov/
56. Reed HH: Circovirus in lories and lorikeets. In Proceedings ahfss/Animal_Health/pdfs/BiosecurityGudelinesforBirdOwners
of the 32nd Joint Annual American Association of Zoo Vet- _END_CDFA-USDA.pdf. (Accessed 17 April 2017).
erinarians/International Association of Aquatic Animals Medical 74. Janssen DL, Sutherland-Smith M, Papendick R, et al: Exotic
Conference, 2000, pp 317–321. Newcastle Disease outbreak in southern California: biosecurity
57. Stalis I, Papendick R: Emphysematous ingluvitis in rainbow measures for prevention in zoo collections. In Proceedings of
lorikeets (Trichoglossus haematodus haemotodus). In Proceedings the 34th Annual American Association of Zoo Veterinarians
of the 47th Annual American Association of Zoo Veterinarians Conference, 2003, pp 107–110.
Conference, 2015, p 136. 75. Steele J: Zoos have plan for avian flu. San Diego Union Tribune.
58. Churria CD, Spinsanti E, Origlia J, et al: Dispharynx nasuta 23 January 2006.
(Nematoda: Acuariidae) infection causing proventicular lesions 76. ZAHP Fusion Center Web site. zahp.aza.org. (Accessed 17
and death in three captive rosellas (Psittaciformes: Psittacidae), March 2017).
J Zoo Wildl Med 42:164–165, 2011. 77. Center for Food Security and Public Health Disinfection
59. Diab SS, Uzal FA, Giannitti F, et al: Cerebrospinal nematodiasis 101 Web site. Characteristics of selected disinfectants.
outbreak in an urban outdoor aviary of cockatiels (Nymphicus Available at: http://www.cfsph.iastate.edu/Disinfection/Assets/
hollandicus) in southern California, J Vet Diag Invest 24:994–999, CharacteristicsSelectedDisinfectants.pdf. (Accessed 4 April
2012. 2017).
60. Galindo-Cardiel I, Opriessnig T, Molina L, et al: Outbreak of 78. Shima AL, Osborn KG: An epornitic of Salmonella typhimurium
mortality in psittacine birds in a mixed-species aviary associated in a collection of lories and lorikeets, J Zoo Wildl Med 20:373–376,
with Erysipelothrix rhusiopathiae infection, Vet Path 49:498–502, 1989.
2012. 79. Seeley KE, Baldanza V, Clayton LA, et al: Diagnosis and success-
61. Palmieri C, Roy P, Dhillon AS, et al: Avian mycobacteriosis ful treatment of trichomonosis in a captive finch population. In
in psittacines: a retrospective study of 123 cases, J Comp Path Proceedings of the 44th Annual American Association of Zoo
148:126–138, 2013. Veterinarians Conference, 2013, pp 103–104.
64
Systemic Isosporosis in Passerine Birds
KAREN A. TERIO AND MICHAEL J. ADKESSON
S
ystemic isosporosis, formerly known as atoxoplasmo- Isospora infect only specific host species or closely related
sis, is a significant disease of captive passerine birds species, and some data suggest that most avian species
worldwide that can cause mortality in all age classes have one or more unique lineages.16 However, genetically
but is of particular concern in hatchling and fledgling birds. divergent parasites can cause disease within the same species
Disease affects a variety of species, and mortality rates may (mixed infections), and the same parasites can be present
be high.1–7 Presence of infection and clinical disease have in genetically divergent hosts.16,23,27–29 Additional research
limited the success of captive breeding and reintroduction into the phylogenetics and host specificity of this group
programs for threatened and endangered passerines, in of parasites is needed to provide clarity on the epidemiol-
particular the Bali mynah (Leucospar rothschildi), which ogy of infection among passerines. It is not certain if only
appears to be particularly susceptible to disease.8,9 Systemic some (and which) species of avian Isospora produce systemic
isosporosis has been diagnosed in a wide variety of free- infections.
ranging passerines worldwide.5,10–18 With notable exceptions Transmission of Isospora is via a fecal-oral route. Asexual
discussed in more detail later, systemic isospora infections in replication (merogony) occurs within intestinal epithelial
wild birds have been asymptomatic or incidental findings at cells followed by gametogony, fertilization, and shedding of
the time of sampling. unsporulated oocysts within feces.30–31 The oocysts sporulate
Systemic isosporosis has been known by numerous within the environment and then become infectious. In
names, including Lankesterella, Toxoplasma, and most the species of Isospora that cause systemic (visceral) disease,
recently Atoxoplasma. Intestinal and extraintestinal forms extraintestinal merogony is common within circulating
were historically thought to be separate parasites, which mononuclear cells and gametogeny may occur, albeit
has contributed to confusion and inconsistent diagnoses. rarely.16,19,29 These circulating cells may serve as a source for
Morphologic studies have suspected that fecal oocysts and reinfection of intestinal epithelial cells and shedding.19,27
extraintestinal merozoites within the same host are due to The life cycle is likely direct, because parasites with
the same parasite.19–22 Molecular characterization of proto- identical genotypes have been identified in all replicative
zoa from different tissues (e.g., liver, spleen, and intestine) phases (gametogony, merogony, and sporogony).16 Vertical
within individual birds has identified identical protozoal transmission has been suggested in blue-crowned laughing
gene sequences within visceral organs and the intestines, thrushes (Dryonastes courtoisi) following diagnosis of the
providing additional evidence that the same parasite may parasite in chicks hatched from disinfected eggs that were
inhabit intestinal and extraintestinal niches.16,23 artificially incubated.7 Insect vectors may serve as paratenic
Differences in morphologic and phylogenetic relatedness hosts, although data are not consistent.10,16,32,33
to other coccidia have further complicated classification.
Systemic Isospora of passerine birds are genetically similar to Clinical Signs and Lesions
Eimeria sp. and have a Stieda body similar to other Eimeria,
unlike mammalian Isospora which lack this structure within Many birds are presumed infected with little to no clinical
their sporocysts. However, they have tetrasporozoic diplospo- signs or outward evidence of disease.3 Difficulties in ante-
rocystic oocysts consistent with Isospora. Recent molecular mortem diagnostics (see later) limit the ability to identify
research has confirmed that Isospora is a polyphyletic clade carriers. When present, clinical signs are generally nonspe-
with two distinct monophyletic groups, those infecting cific and may include ruffled feathers, dyspnea, tachypnea,
mammals and those infecting birds.16,24 Because Isospora is anorexia, hyporexia, weight loss, diarrhea, dehydration, and
the oldest name, Atoxoplasma (Garnham 1950) is now con- death. Juvenile and fledgling birds are most susceptible, but
sidered a junior synonym of the genus Isospora (Schneider disease may also occur in adults.
1881).24–26 There are multiple described and named species, The characteristic gross lesion of systemic isosporosis is
plus even more that have not been thoroughly classified splenomegaly; however, this finding is variable. In some
beyond Isospora sp. It has been hypothesized that individual cases, small tan foci of necrosis and/or inflammation may
454
CHAPTER 64 Systemic Isosporosis in Passerine Birds 455
• Figure 64.1 Diff-Quik–stained impression smear of spleen from • Figure 64.3 Hematoxylin-eosin–stained histologic section of heart
an amakihi (Chlorodrepanis virens). Note Isospora sp. within the from a red-vented bulbul (Pycnonotus cafer) with myositis associ-
cytoplasm of mononuclear cells indenting the nucleus (arrows). ated with systemic isosporosis. Myocytes are fragmented (examples
highlighted with asterisks) and surrounded by inflammatory cells.
• Figure 64.2 Hematoxylin-eosin–stained histologic section of spleen • Figure 64.4 Hematoxylin-eosin–stained histologic section of intes-
from a Taveta golden weaver (Ploceus castaneiceps) with lympho- tine from a Taveta golden weaver (Ploceus castaneiceps). Multiple
histiocytic splenitis associated with systemic isosporosis. Increased Isospora sp. microgamonts (arrows) are present within the intestinal
numbers of lymphocytes and histiocytes are present surrounding epithelium.
splenic ellipsoids or Schweigger-Seidel sheaths (examples highlighted
with asterisks). with other systemic diseases, any organ may be potentially
infected. Infected mononuclear cells can be adherent to
be noted in the spleen and liver. Other lesions may include blood vessel endothelium within the lung.4 Cytoplasmic
thickened and/or dilated intestines, an enlarged darkened merozoites may be difficult to visualize within histologic
liver (“black spot” because it can be viewed through the body sections, and inflammation alone is nonspecific; therefore
wall), and pale streaks within the heart and skeletal muscle. impression smears are critical for diagnosis. In one case
On splenic, lung, or liver impression smears stained with series a captive population of red-vented bulbuls (Pycnono-
Wright-Giemsa (or other Romanowsky-type stains such tus cafer) had significant and fatal skeletal and myocardial
as Diff-Quik), lymphocytes and histiocytes may contain lesions (Fig. 64.4) in the absence of characteristic splenic and
one to multiple 2–3 µm oval-shaped intracytoplasmic hepatic lesions.23 In American goldfinches (Spinus tristus)
pale basophilic to eosinophilic merozoites surrounded by and house sparrows (Passer domesticus), T-cell lymphocytic
a colorless capsule (parasitophorous vacuole) that indents infiltrates with cytoplasmic protozoal meronts effacing the
the nucleus (Fig. 64.1). normal small intestinal mucosa and infiltrating through the
Histologically, necrosis and/or lymphoid, histiocytic, or intestinal wall into the coelomic cavity suggestive of a T-cell
lymphohistiocytic inflammation is common (Figs. 64.2 and lymphoma have been reported.6 However, further research
64.3).1,2,4–6,8,10,29,34–37 Inflammation is often in vascular and/ is needed to demonstrate the oncogenic potential of the
or perivascular spaces. The most commonly affected sites parasite. Other immunohistochemical studies have identi-
include spleen, liver, heart, lung, and intestine; however, as fied the majority of parasitized cells as B lymphocytes.35
456 SE C T I O N 12 Avian
TABLE
64.1 Reported Treatment Protocols for Reducing the Shedding of Systemic Isosporosis in Passerines
Some infected birds have no associated gross or histologic However, impression smears are not useful as a screen-
lesions. In studies of systemic Isospora in wild birds, clinical ing tool in live birds. Cytologic evaluation of whole blood
manifestations were lacking in most cases. Unrecognized smears and buffy coat smears may identify merozoites within
stressors in artificial environments may precipitate disease in peripheral mononuclear leukocytes; however, organisms
previously infected wild birds. In one study of wild-caught can be rare even in fulminant, fatal infections.5 Consistent
passerines, approximately half of the captured birds died complete blood count (CBC) and chemistry findings are
within 15 days of confinement, many of which had systemic lacking, and alterations may represent secondary condi-
isosporosis, although they were also concurrently infected tions such as dehydration. Organisms may be identified in
with Leukocytozoon (Gill et al., 2008).5 Concurrent Leuko- feces by routine floatation followed by sporulation, if they
cytozoon infection may also have impacted disease develop- are shed within the sample. Because shedding is intermit-
ment in free-ranging rose-breasted grosbeaks (Pheucticus tent, false negatives may occur. Challenges of antemortem
ludovicianus) with systemic visceral infection.10 diagnosis in juvenile passerines are compounded by limited
access to diagnostic samples due to small size of the animal
Diagnosis and disruption of the next. Polymerase chain reaction
(PCR)-based assays are more likely to be of utility due
The “gold standard” for diagnosis remains impression smears to increased sensitivity, although, because fecal shedding
of visceral organs, specifically spleen, liver, and lung. Within and systemic phases of disease can be intermittent, studies
these impressions, two separate stages may be identified— of shedding patterns are needed to determine optimal
larger single merozoites within a single parasitophorous sampling intervals.3,16,39
vacuole (“waiting” merozoite) and multiple smaller mero-
zoites sharing a common parasitophorous vacuole (“pro- Treatment and Management
liferative” merozoites).5,27,36–38 It is the merozoite within
mononuclear cells that causes the characteristic appearance Antiprotozoal drugs are used for both treatment of clinical
of an “indentation” in the nucleus (see Fig. 64.1). disease and suppression of shedding (Table 64.1). Extensive
CHAPTER 64 Systemic Isosporosis in Passerine Birds 457
study on therapeutic dosages and treatment efficacy is as Isospora sp. and have both intestinal and extraintestinal
lacking. Treatment is generally aimed at reducing shedding phases. Lesions vary but classically are associated with lym-
during breeding season through chick fledging. Sulfonamide phohistiocytic inflammation. Differing disease manifesta-
drugs (e.g., sulfachloropyrazine, sulfachlorpyridazine, sul- tions may be due to differences in host-parasite interactions
fadimethoxine) in the water are reported to reduce disease among species of Isospora, as well as species of passerine.
by limiting shedding, but controlled studies are lacking and Current gold-standard diagnostics (postmortem impres-
purported efficacy is based primarily on decreases in chick sion smears) are unrewarding for clinical management, but
mortality. Toltrazuril has emerged as an effective treatment the advent of molecular tools will allow more in-depth
for reducing shedding of oocysts, although the drug is not studies into the epidemiology and disease pathogenesis so
commercially available in the United States.7,40 Ponazuril that management and treatment protocols can be further
has been used to reduce shedding and successfully treat improved.
clinical cases, although controlled studies of this drug are
also lacking. Treatment can effectively eliminate shedding
and reduce intestinal and systemic stages, but elimination References
of infection is unlikely.
Husbandry and sanitation are extremely important 1. Cooper JE, Gschmeissner S, Greenwood AG: Atoxoplasma in
components of any plan for controlling systemic isosporosis greenfinches (Carduelis chloris) as a possible cause of “going
light”, Vet Rec 124:343–344, 1989.
infections in an aviary setting, particularly with breeding
2. Swayne DE, Getzy D, Slemons RD, et al: Coccidiosis as cause of
birds. Substrates should be cleaned on a regular basis, and transmural lymphocytic enteritis and mortality in captive Nash-
fecal contamination of food and water sources should ville warblers (Vermivora ruficapilla), J Wildl Dis 27:615–620,
be minimized through the use of elevated, covered feed 1991.
stations. Minimizing stress and disturbance of breeding 3. Adkesson MJ, Zdiarski JM, Little SE: Atoxoplasmosis in tanagers,
birds may aid in minimizing shedding and infection. Pro- J Zoo Wildl Med 36:265–272, 2005.
phylactic treatment with antiprotozoal medications during 4. Sánchez-Cordón PJ, Gómez-Villamandos JC, Gutiérrez J, et al:
times of stress has been proposed, but the efficacy of such Atoxoplasma spp. infection in captive canaries (Serinus canaria),
intervention is not known. Regular diagnostic testing of J Vet Med A Physiol Pathol Clin Med 54:23–26, 2007.
feces may also aid in identification of individual birds that 5. Gill H, Paperna I: Proliferative visceral Isospora (atoxoplasmosis)
are chronically shedding at high levels so they can be treated with morbid impact on the Israeli sparrow Passer domesticus
biblicus Hartert, 1904, Parasitol Res 103:493–499, 2008.
or removed from breeding populations.
6. Cushing TL, Schat KA, States SL, et al: Characterization of
If high levels of chick mortality are present from systemic the host response in systemic Isosporosis (Atoxoplasmosis) in a
isosporosis, breeding adults should be administered antipro- colony of captive American goldfinches (Spinus tristis) and house
tozoal medications to reduce shedding. Treatment should sparrows (Passer domesticus), Vet Pathol 48:985–992, 2011.
commence prior to egg laying and continue in pulsed form 7. Mohr F, Betson M, Quintard B: Investigation of the presence
while the parents continue to feed fledglings. The physio- of Atoxoplasma spp. in blue-crowned laughing thrush (Dyro-
logic stress of fledging likely predisposes chicks to infec- nases courtoisi) adults and neonates, J Zoo Wildl Med 48:1–6,
tion, and earlier cessation of treatment may lead to disease 2017.
because fecal shedding of oocysts has been shown to return 8. Partington CJ, Gardiner CH, Fritz D, et al: Atoxoplasmosis in
to pretreatment levels within as little as 10 days.40 However, Bali mynahs (Leucospar rothschildi), J Zoo Wildl Med 20:328–335,
untested negative correlations with clutch size and laying 1989.
9. Done LB, Case JT, Mazet JA, et al: Assessment of the quality
frequency have been reported with toltrazuril at higher
of multi-institutional data for population-based epidemiological
dosages.7 The toxicity, side effects, appropriate dosage, and studies: a case study of atoxoplasmosis in Bali mynahs (Leucospar
efficacy of various treatments are areas that warrant signifi- rothschildi), J Zoo Wildl Med 42:228–235, 2011.
cant investigation. Advances in molecular testing are allow- 10. Khan RA, Desser SS: Avian Lankesterella infections in Algonquin
ing investigation of shedding patterns and offer promise for Park, Ontario, Can J Zool 49:1105–1110, 1971.
better monitoring and assessment of treatment efficacy. Dis- 11. Peirce MA: Distribution and host-parasite check-list of the
infection of eggs and artificial incubation may be indicated hematozoa of birds in western Europe, J Nat Hist 15:419–458,
in certain situations, although it is unknown if a lack of 1981.
exposure to the organism early in life may actually increase 12. Bennett GF, Garvin M, Bates JM: Avian hematozoa from west-
susceptibility to clinical disease later in life. As noted previ- central Bolivia, J Parasitol 77:207–211, 1991.
ously, vertical transmission may also be possible. 13. Bennett GF, Earle RA, Du Toit H, et al: A host-parasite catalogue
of the haematozoa of the sub-Saharan birds, Onderstepoort J Vet
Res 59:1–73, 1992.
Conclusion 14. Pennycott TW, Ross HM, McLaren IM, et al: Causes of death
of wild birds of the family Fringillidae in Britain, Vet Rec
Conservation and breeding programs for passerines require 143:155–158, 1998.
improved medical management of systemic isosporosis. 15. Valkiunas G, Iezhova TA: A comparison of the blood parasites
Despite considerable prior confusion over phylogeny (e.g., in three subspecies of yellow wagtail Motacilla flava, J Parasitol
Atoxoplasma, Lankesterella), these parasites are now classified 87:930–934, 2001.
458 SE C T I O N 12 Avian
16. Schrenzel MD, Maalouf GA, Gaffney PM, et al: Molecular 29. Hafeez MA, Stasiak I, Delnatte P, et al: Description of two new
characterization of Isosporoid coccidia (Isospora and Atoxoplasma Isospora species causing visceral coccidiosis in captive superb
spp.) in passerine birds, J Parasitol 91:635–647, 2005. glossy starlings, Lamprotornis superbus (Aves: Sturnidae), Parasitol
17. Valkiunas G, Sehgal RN, Iezhova TA, et al: Further observa- Res 113:3287–3297, 2014.
tions on the blood parasites of birds in Uganda, J Wildl Dis 30. Box ED: Atoxoplasma associated with an isosporan oocyst in
41:580–587, 2005. canaries, J Protozool 17:391–396, 1970.
18. Lindstrom KM, Dolnik O, Yabsley M, et al: Feather mites and 31. Upton SJ, Wilson SC, Norton TM, et al: A new species of
internal parasites in small ground finches (Geospiza fuliginosa, Isospora Schneider, 1881 (Apicomplexa: Eimeriidae) from the
Emberizidae) from the Galapagos Islands (Ecuador), J Parastiol. Bali (Rothschild’s) Mynah Leucopsar rothschildi (Passeriformes:
95:39–45, 2009. sturnidae) and comments concerning the genera Atoxoplasma
19. Box ED: Life cycles of two Isospora species in the canary, Serinus Garnham, 1950 and Isospora, System Parasitol 48:47–53,
canaries Linnaeus, J Protozool 24:57–67, 1977. 2001.
20. Grulet O, Landau I, Millet P, et al: Les Isospora du moineau. II 32. Lainson R: Some observations on the life cycle of Atoxoplasma,
études sur la biologie, Annal Parsitol Hum Comp 61:161–192, with particular reference to the parasite’s schizogony and
1986. its transmission by the mite Dermanyssus gallinae, Nature
21. Boulard Y, Landau I, Grulet O, et al: Ultrastructure of chronic 182:1250–1251, 1958.
reticuloendothelial forms of Isospora of sparrows, Ann Parasitol 33. Lainson R: The transmission of Lankesterella (=Atoxoplasma) in
Hum Comp 62:181–184, 1987. birds by the mite Dermanyssus gallinae, J Protozool 7:321–322,
22. Ball SJ, Brown MA, Daszak P, et al: Atoxoplasma (Apicomplexa: 1960.
Eimeriorina: Atoxoplasmatidae) in the greenfinch (Carduelis 34. Martinez F, Munoz E: Atoxoplasma spp. in a hybrid pas-
chloris), J Parasitol 84:813–817, 1998. serine (Serinus canarius x Carduelis cannabina), Avian Pathol
23. Righton A, Adkesson M, Snyder T, et al: Systemic isosporosis 27:420–422, 1998.
(Isospora sp.) associated with myocarditis and myositis in pas- 35. Maslin WR, Latimer KS: Atoxoplasmosis in canary fledglings:
serine birds, unpublished manuscript, 2017. severe lymphocytic enteritis with preferential parasitism of B
24. Barta JR, Schrenzel MD, Carreno R, et al: The Genus Atoxo- lymphocytes, Avian Dis 53:473–476, 2009.
plasma (Garnham 1950) as a junior objective synonym of the 36. Middleton ALA, Julian RJ: Lymphoproliferative disease in the
Genus Isospora (Schneider 1881) species infecting birds and American goldfinch, Carduelis tristis, J Wildl Dis 19:280–285,
resurrection of Cystisospora (Frenkel 1977) as the correct genus 1983.
for Isospora species infecting mammals, J Parasitol 91:726–727, 37. McNamee P, Pennycott T, McConnel S: Clinical and pathologi-
2005. cal changes associated with atoxoplasma in a captive bullfinch
25. Garnham PCC: Blood parasites of east African vertebrates with (Pyrrhula pyrrhula), Vet Rec 136:221–222, 1995.
a brief description of exo-erythrocytic schizogony of P. pitmani, 38. Desser SS: An ultrastructural study of the asexual development
Parasitol 40:328–337, 1950. of a presumed Isospora sp. in mononuclear, phagocytic cells
26. Schneider A: Sur les psorospermies oviformes ou coccidies. of the evening grosbeak (Hesperiphona vespertina), J Parasitol
Especes nouvelles ou peu connues, Arch Zool Exp Gen 9:387–404, 66:601–612, 1980.
1881. 39. Landolfi J, Adkesson MJ, Snyder T, et al: Development of a
27. Milde K: Light and electron microscopic studies of isosporan qPCR for Atoxoplasma diagnosis in passerine birds. In Proceed-
parasites (Sporozoa) in sparrows (Passer domesticus L.), Protistol ings of the Joint Conference of American Association of Zoo
15:607–627, 1979. Veterinarians/EAZWV/IZW 2016, p 61.
28. Grulet O, Landau I, Baccam D: Isospora from the domestic 40. Jamriška J, Lavilla LA, Thomasson A, et al: Treatment of
sparrow; multiplicity of species, Ann Parasitol Hum Comp 57: atoxoplasmosis in the blue-crowned laughing thrush (Dryonastes
209–235, 1982. courtoisi), Avian Pathol 42:569–571, 2013.
65
Bornaviruses in Birds
DALE SMITH
T
he discovery of the first avian bornavirus in 2008 free-ranging psittacine birds is from Brazil, where seroposi-
provided the key to solving a mystery that had been tivity and shedding of bornaviral RNA were identified in
plaguing avian veterinarians since the late 1970s, parrots, including nestlings, recently taken into rehabilita-
when macaw wasting disease, as it was then called, was first tion centers.8 Preliminary research suggests that there are
identified in macaws imported into the United States from differences in pathogenesis among the different parrot
Bolivia.1–3 This disease, now most commonly known as bornaviruses, but the relevance of this to managing clinical
proventricular dilation (dilatation) disease (PDD), became situations is not yet clear.9
the most feared and least understood infection of captive Two species of bornavirus affect passerine birds. Canar-
psittacine birds. Although there has been debate in the ies (Serinus canaria) infected with canary bornaviruses
veterinary community regarding the relationship between 1-3 (Passeriform 1 bornavirus) can show PDD-like clinical
bornaviral infection and PDD, researchers working with signs and pathologic lesions.10,11 Experimental infection
this group of viruses are convinced the relationship is did not result in clinical disease or significant pathology;
causal, although complex. For clarity, the term PDD is however, horizontal transmission to in-contact birds did
used to describe cases showing the classic disease syndrome, occur.10 Twelve of 30 German canary flocks were positive
based on clinical and/or pathologic features, and bornaviral for virus, suggesting a high prevalence of disease in canaries
infection is confirmed by the presence of viral RNA, viral in that country.10 Passeriform 2 bornavirus (estreldid finch
antigen, or viable virus. bornavirus 1, EsBV-1) was described in 2014 from three
Prior to the discovery of this first avian bornavirus, estreldid finches in a single flock in Germany as part of an
the only member of the family Bornaviridae was Borna extensive screening program.12 Because there were a number
disease virus (BDV), a cause of neurologic disease in of disease processes ongoing in the flock, the significance of
mammals in central Europe, particularly horses and sheep.4 EsBV-1 could not be determined.
Bornaviruses are members of the order Mononegavirales An investigation into the cause of encephalitis of
and are enveloped viruses characterized by the presence of unknown origin in Canada geese (Branta canadensis) and
nonsegmented, linear, single-stranded negative-sense RNA trumpeter (Cygnus buccinator) and mute swans (Cygnus olor)
genomes. Bornavirus infections are often persistent and are in Canada led to the discovery of Waterbird 1 bornavirus
noncytopathic in cell culture; hence the need for molecular (aquatic bird bornavirus 1, ABBV-1).13 Histopathologic
diagnostic techniques in viral discovery. lesions were similar to those of PDD, as were gross necropsy
Since 2008 there has been an explosion in the discovery findings and clinical history in the birds for which this
of new members of the Bornaviridae, particularly those information was available.14 A prospective serologic study in
affecting avian species (Table 65.1).5 Additional viral geno- these species by the same authors and extensive surveillance
types await formal classification. Several excellent reviews by researchers in the United States showed that antibodies
summarize knowledge current to their publication dates.2,6,7 and viral RNA were present at very high prevalence in
Active research is ongoing, and thus each year brings new seemingly unaffected birds, as well as in culled, apparently
insights into the epidemiology and pathogenesis of avian healthy gulls of several species.15–17 Infection with ABBV-1
bornaviral infections. thus appears to be widespread in free-ranging waterfowl and
PDD is recognized worldwide in a broad range of psit- gulls in North America, but associated with disease only in
tacine species; global spread is thought to have occurred a subset of cases. ABBV-1 viral RNA was also identified
through the trade in captive birds. Disease has been most in the brains of hunter-killed free-ranging waterfowl in
frequently associated with infection by parrot bornavirus Denmark.18 ABBV-1 was identified in the embryos of Pekin
2 (PaBV-2) and 4 (PaBV-4). The natural reservoir (i.e., ducks in eggs purchased commercially in the United States
the source of entry of virus into the captive population) for laboratory use; however, there are no associated surveys
is not known. The only report of bornaviral infection in of commercial duck flocks.7
459
460 SE C T I O N 12 Avian
TABLE Members of the Family Bornaviridae, Genus Although BDV is recognized as a pathogen of mammals,
65.1 Bornavirus, Associated With Disease in BDV RNA was found in the feces of mallards and jackdaws
Avian Species (Corvus monedula) in Sweden.22 Borna disease virus, or a
very similar virus, caused a paralytic syndrome with high
Species Virus Abbreviation mortality in young ostriches (Struthio camelus) in Israel.23
Passeriform 1 Canary bornavirus 1 CnBV-1 Both reports predated the identification of the first avian
bornavirus Canary bornavirus 2 CnBV-2 bornavirus and, in the ostrich outbreak, the use of RT-PCR
Canary bornavirus 3 CnBV-3 and sequencing as tools for virus identification.
Passeriform 2 Estrildid finch EsBV-1
bornavirus bornavirus Antemortem Clinical Diagnosis
Psittaciform 1 Parrot bornavirus 1 PaBV-1
bornavirus Parrot bornavirus 2 PaBV-2 Diagnosing the clinical condition known as PDD and con-
Parrot bornavirus 3 PaBV-3 firming the presence of bornaviral infection in the live bird
Parrot bornavirus 4 PaBV-4 are not always clear-cut. Infection and disease can occur
Parrot bornavirus 7 PaBV-7
in birds of all ages. The classic clinical signs in psittacine
Psittaciform 2 Parrot bornavirus 5 PaBV-5 birds are weight loss, often with continuing appetite; the
bornavirus passage of poorly ground or undigested food items, such
Waterbird 1 Aquatic bird ABBV-1 as seeds and nuts; and neurologic abnormalities. However,
bornavirus bornavirus 1 birds may have nonspecific signs of ill health, show some
Aquatic bird ABBV-2 but not all of the previous signs, or be subclinically affected
bornavirus 2
with no overt clinical signs. There are also reports of sudden
From Amarasinghe GK, Bào Y, Basler CF, et al: Taxonomy of the order death as a result of myocardial disease and of blindness. The
Mononegavirales: update 2017. Arch Virol 162:1–12, 2017. development and course of clinical disease in an infected
parrot is completely unpredictable, in terms of both time to
onset and duration of clinical signs. Parrots have developed
A second waterfowl-associated bornavirus, aquatic bird clinical PDD years after contact with any other bird.6 There
bornavirus 2 (ABBV-2), was identified from the brains of are no consistent patterns in complete blood count or
hunter-killed mallards (Anas platyrhynchos) and a wood plasma/serum biochemical profiles; a transient increase in
duck (Aix sponsa) from Oklahoma, United States.19 Viral gamma globulins was seen in African grey parrots (Psittacus
antigen was present in retina and in brain, where it was erithacus) experimentally infected with PaBV-4.24 Imaging
accompanied by mild perivascular cuffing and gliosis in the findings that support a diagnosis of PDD include dilation
few specimens examined histologically. The significance and of the crop, proventriculus, or ventriculus; and reduced or
prevalence of ABBV-2 are unknown. abnormal gastrointestinal motility as viewed on contrast
The bornaviruses affecting birds are quite consistently fluoroscopic examination (Fig. 65.1).
associated with given taxonomic groups, but there are The gold standard for diagnosis of PDD in a live bird has
some notable exceptions. ABBV-1 was identified by reverse historically been the observation of a lymphoplasmacytic
transcription polymerase chain reaction (RT-PCR) from ganglioneuritis in a biopsy of the crop wall. This technique
the brain of a bald eagle (Haliaeetus leucocephalus) with has poor sensitivity because biopsy specimens vary con-
encephalitis, the presumption being that the eagle became siderably in the number of nerves and ganglia present for
infected by eating infected waterfowl.2 ABBV-1 was simi- evaluation, and lesions vary in their distribution.25,26 The
larly identified, using immunohistochemistry (IHC) and application of IHC and RT-PCR to either fresh, frozen, or
RT-PCR, in the brain of a captive emu (Dromaius novaehol- formalin-fixed paraffin-embedded (FFPE) tissue to identify
landiae) with encephalitis and lymphoplasmacytic infiltrates the present of bornaviral RNA or antigens increases the sen-
in autonomic ganglia of the gastrointestinal tract.20 The sitivity and specificity of diagnosis based on crop biopsy.27
bird’s outdoor enclosure was frequented by Canada geese in Many veterinary diagnostic laboratories offer either con-
a population shown to have a high prevalence of ABBV-1 ventional (gel-based) or real-time (quantitative) RT-PCR
infection; fecal-oral transmission was proposed. PaBV-4 was that targets, most commonly, the gene for the matrix (M)
identified in a Himalayan monal (Lophophorus impejanus) protein or the nucleoprotein (N). Sequencing of products is
with clinical signs and pathologic lesions consistent with required to determine the specific virus present. Bornaviral
PDD that shared an aviary with parrots.21 PDD had previ- RNA is present in choanal-cloacal, crop, and cloacal swabs;
ously been identified in psittacines in the collection, and feces; and urine of infected birds.6,27,28 However, shedding
PaBV-4 was subsequently identified from a diseased white- is intermittent, particularly in cloacal swabs and feces, and
bellied caique (Pionites leucogaster). The avian bornaviruses thus false-negative results may be common.6,27 It has been
should thus be considered in the differential of any case of suggested that bornaviral RNA is present in the feather
nonsuppurative encephalitis in a bird, particularly when calami of infected birds, that plucked, mature contour
appropriate lesions are present in other components of the feathers can be used as diagnostic material, and that this
nervous system. material is stable for at least 4 weeks at room temperature.29
CHAPTER 65 Bornaviruses in Birds 461
Treatment References
There are two aspects of treatment of birds with bornavirus 1. Honkavuori KS, Shivaprasad HL, Williams BL, et al: Novel
infection: reducing or clearing viral load and ameliorating Borna virus in psittacine birds with proventricular dilatation
clinical signs. Ribavirin inhibits replication of PaBV-4 in disease, Emerging Infect Dis 14:1883–1886, 2008.
cell culture, but preliminary pharmacokinetics indicated 2. Hoppes SM, Tizard I, Shivaprasad HL: Avian bornavirus
and proventricular dilatation disease: diagnostics, pathology,
it would be difficult to obtain and maintain therapeutic
prevalence, and control, Vet Clin North Am Exot Anim Pract
levels.42,43 Favipirivir (T-705) strongly suppressed the 16:339–355, 2013.
growth of PaBV-4 in cell culture; this drug is licensed for 3. Kistler AL, Gancz A, Clubb S, et al: Recovery of divergent avian
oral use in humans, but there are no reports of its in vivo bornaviruses from cases of proventricular dilatation disease:
use in birds.44 identification of a candidate etiologic agent, Virol J 5:88,
Drugs targeting inflammation appear to be the most 2008.
logical treatment to reduce clinical signs, which are assumed 4. Tizard I, Ball J, Stoica G, et al: The pathogenesis of bornaviral
to result at least in part from an immune reaction to the diseases in mammals, Anim Health Res Rev 17:92–109, 2016.
presence of the virus. Treatment with celecoxib (10 mg/kg 5. Amarasinghe GK, Bào Y, Basler CF, et al: Taxonomy of the order
by mouth, once daily) has been reported to dramatically Mononegavirales: update 2017, Arch Virol 162:1–12, 2017.
reduce clinical signs in some birds with PDD.45 Unex- 6. Lierz M: Avian bornavirus and proventricular dilation disease. In
Speer BL, editor: Current therapy in avian medicine and surgery.
pectedly, more severe disease was noted in experimentally
St. Louis, 2016, Elsevier, pp 28–46.
infected cockatiels treated with meloxicam, and there is 7. Payne SL, Delnatte P, Guo J, et al: Birds and bornaviruses, Anim
little evidence of improvement using cyclosporine.2,46 Health Res Rev 13:145–156, 2012.
General supportive care, including the provision of an easily 8. Encinas-Nagel N, Enderlein D, Piepenbring A, et al: Avian
digested diet, can often improve the general well-being and bornavirus in free-ranging psittacine birds, Brazil, Emerging Infect
body condition of birds showing clinical signs of PDD and Dis 20:2103–2106, 2014.
extend their life span; however, the potential for transmis- 9. Al-Ibadi B, Piepenbring A, Herzog S, et al: Comparison between
sion of virus to uninfected birds must always be considered. ABV2 and ABV4 distribution patterns in experimentally infected
cockatiels, J Comp Pathol 152:52, 2012.
10. Rubbenstroth D, Rinder M, Stein M, et al: Avian bornaviruses
Control are widely distributed in canary birds (Serinus canaria f. domes-
tica), Vet Microbiol 165:287–295, 2013.
Effective quarantine protocols and separation of infected
11. Weissenbock H, Sekulin K, Bakonyi T, et al: Novel avian bor-
and uninfected birds remain the sole methods of preventing navirus in a nonpsittacine species (canary; Serinus canaria) with
the horizontal spread of bornavirus infection. No specific enteric ganglioneuritis and encephalitis, J Virol 83:11367–11371,
information is available on the environmental survival or 2009.
sensitivity to disinfectants for the avian bornaviruses; they 12. Rubbenstroth D, Schmidt V, Rinder M, et al: Discovery of a
are assumed to have the same stability as other similar new avian bornavirus genotype in estrildid finches (Estrildidae)
enveloped RNA viruses. Disinfection protocols using in Germany, Vet Microbiol 168:318–323, 2014.
hypochlorites (bleach), phenols, and formaldehyde are 13. Delnatte P, Berkvens C, Kummrow M, et al: New genotype of
suggested. avian bornavirus in wild geese and trumpeter swans in Canada,
Given the difficulties in identifying infected birds, Vet Rec 169:108, 2011.
particularly in the early stages of infection or when there 14. Delnatte P, Ojkic D, Delay J, et al: Pathology and diagnosis of
avian bornavirus infection in wild Canada geese (Branta canaden-
is a chronic carrier state, prevention of new infection and
sis), trumpeter swans (Cygnus buccinator) and mute swans (Cygnus
elimination of disease within a flock remain problematic. olor) in Canada: a retrospective study, Avian Pathol 42:114–128,
Combined use of RT-PCR evaluation of cloacal swabs 2013.
and serology is recommended, with repeated cycles of 15. Delnatte P, Nagy É, Ojkic D, et al: Avian bornavirus in free-
testing and separation of infected, possibly infected, and ranging waterfowl: prevalence of antibodies and cloacal shedding
apparently uninfected birds until at least two sequential of viral RNA, J Wildl Dis 50:512–523, 2014.
negative results are obtained for the presumed negative 16. Guo J, Covaleda L, Heatley JJ, et al: Widespread avian bornavi-
birds.6 Collection of eggs for artificial incubation and hand- rus infection in mute swans in the northeast United States, Vet
raising chicks in a clean facility have been proposed as Med Res Reports 3:49–52, 2012.
components of conservation programs dealing with endan- 17. Guo J, Tizard I, Baroch J, et al: Avian bornaviruses in North
gered species, with awareness of the possibility of vertical American gulls, J Wildl Dis 51:754–758, 2015.
18. Thomsen AF, Nielsen JB, Hjulsager CK, et al: Aquatic bird
transmission.47
bornavirus 1 in wild geese, Denmark, Emerging Infect Dis
Research into the development of vaccines is ongoing. 21:2201–2203, 2015.
Recently, vectored virus vaccines expressing N protein 19. Guo J, Shivaprasad HL, Rech RR, et al: Characterization of
and phosphoprotein genes from PaBV-4 provided partial a new genotype of avian bornavirus from wild ducks, Virol J
protection against experimental inoculation with PaBV-2 11:197, 2014.
in cockatiels.48 An effective, widely available vaccine is still 20. Nielsen AMW, Ojkic D, Dutton CJ, et al: Aquatic bird borna-
a long way from reality. virus 1 infection in a captive emu (Dromaius novaehollandiae):
464 SE C T I O N 12 Avian
presumed natural transmission from free-ranging wild waterfowl, reverse transcription polymerase chain reaction, J Vet Diagn Invest
Avian Pathol 47(1):58–62, 2018. 26:266–271, 2014.
21. Bourque L, Laniesse D, Beaufrère H, et al: Identification of avian 35. Rubbenstroth D, Rinder M, Kaspers B, et al: Efficient isolation
bornavirus in a Himalayan monal (Lophophorus impejanus) with of avian bornaviruses (ABV) from naturally infected psittacine
neurological disease, Avian Pathol 44:323–327, 2015. birds and identification of a new ABV genotype from a salmon-
22. Berg M, Johansson M, Montell H, et al: Wild birds as a pos- crested cockatoo (Cacatua moluccensis), Vet Microbiol 161:36–42,
sible natural reservoir of Borna disease virus, Epidemiol Infect 2012.
127:173–178, 2001. 36. Tizard I, Shivaprasad HL, Guo J, et al: The pathogenesis of pro-
23. Malkinson M, Weisman Y, Perl S, et al: A Borna-like disease of ventricular dilatation disease, Anim Health Res Rev 17:110–126,
ostriches in Israel. In Koprowski H, Lipkin W, editors: Curr Top 2016.
Microbiol Immunol 190:31–38, 1995. 37. Kistler AL, Smith JM, Greninger AL, et al: Analysis of naturally
24. Högemann C, Richter R, Korbel R, et al: Plasma protein, occurring avian bornavirus infection and transmission during
hematologic and blood chemistry changes in African grey parrots an outbreak of proventricular dilatation disease among captive
(Psittacus erithacus) experimentally infected with bornavirus, psittacine birds, J Virol 84:2176–2179, 2010.
Avian Pathol 46:1–70, 2017, doi:10.1080/03079457.2017.132 38. Lublin A, Mechani S, Farnoushi I, et al: An outbreak of proven-
5442. tricular dilatation disease in psittacine breeding farm in Israel, Isr
25. Berhane Y, Smith DA, Newman S, et al: Peripheral neuritis in J Vet Med 61:16–19, 2006.
psittacine birds with proventricular dilatation disease, Avian 39. Heckmann J, Enderlein D, Piepenbring AK, et al: Investigation
Pathol 30:563–570, 2001. of different infection routes of parrot bornavirus in cockatiels,
26. Gregory CR, Latimer KS, Campagnoli RP, et al: Histologic Avian Dis 61:90–95, 2017.
evaluation of the crop for diagnosis of proventricular dilatation 40. Wüst E, Mahlberg S, Enderlein D, et al: Experimentally infec-
syndrome in psittacine birds, J Vet Diagn Invest 8:76–80, 1996. tion of cockatiel (Nymphicus hollandicus) eggs. In 2nd ICARE
27. Raghav R, Taylor M, Delay J, et al: Avian bornavirus is present in Conference. Paris, France: ICARE, 2015: 258.
many tissues of psittacine birds with histopathologic evidence of 41. Monaco E, Hoppes S, Guo J, et al: The detection of avian bor-
proventricular dilatation disease, J Vet Diagn Invest 22:495–508, navirus within psittacine eggs, J Avian Med Surg 26:144–148,
2010. 2012.
28. Heatley J, Villalobos AR: Avian bornavirus in the urine of 42. Hoppes S, Gray PL, Payne S, et al: The isolation, pathogenesis,
infected birds, Vet Med Res Reports 3:19–23, 2012. diagnosis, transmission, and control of avian bornavirus and
29. de Kloet AH, Kerski A, de Kloet SR: Diagnosis of avian bor- proventricular dilatation disease, Vet Clin North Am Exot Anim
navirus infection in psittaciformes by serum antibody detection Pract 13:495–508, 2010.
and reverse transcription polymerase chain reaction assay using 43. Musser JMB, Heatley JJ, Koinis AV, et al: Ribavirin inhibits
feather calami, J Vet Diagn Invest 23:421–429, 2011. parrot bornavirus 4 replication in cell culture, PLoS ONE
30. Guo J, Payne S, Zhang S, et al: Avian bornaviruses: diagnosis, 10:1–17, 2015.
isolation, and genotyping, Curr Protoc Microbiol 34:15I.1.1 44. Tokunaga T, Yamamoto Y, Sakai M, et al: Antiviral activity of
–15I.1.33, 2014. favipiravir (T-705) against mammalian and avian bornaviruses,
31. Kerski A, de Kloet AH, de Kloet SR: Vertical transmission of Antiviral Res 143:237–245, 2017.
avian bornavirus in Psittaciformes: avian bornavirus RNA and 45. Dahlhausen B, Aldred S, Colaizzi E: Resolution of clinical
anti-avian bornavirus antibodies in eggs, embryos, and hatchlings proventricular dilatation disease by cyclooxygenase 2 inhibition,
obtained from infected sun conures (Aratinga solstitialis), Avian Assoc Avian Vet:9–12, 2002.
Dis 56:471–478, 2012. 46. Hoppes S, Heatley JJ, Guo J, et al: Meloxicam treatment in
32. Zimmermann V, Rinder M, Kaspers B, et al: Impact of antigenic cockatiels (Nymphicus hollandicus) infected with avian bornavi-
diversity on laboratory diagnosis of avian bornavirus infections rus, J Exot Pet Med 22:275–279, 2013.
in birds, J Vet Diagn Invest 26:769–777, 2014. 47. Lierz M, Piepenbring A, Herden C: Vertical transmission of avian
33. Rinder M, Ackermann A, Kempf H, et al: Broad tissue and bornavirus in psittacines, Emerging Infect Dis 17:2390–2391,
cell tropism of avian bornavirus in parrots with proventricular 2011.
dilatation disease, J Virol 83:5401–5407, 2009. 48. Runge S, Olbert M, Herden C, et al: Viral vector vaccines protect
34. Delnatte P, Mak M, Ojkic D, et al: Detection of avian bornavirus cockatiels from inflammatory lesions after heterologous parrot
in multiple tissues of infected psittacine birds using real-time bornavirus 2 challenge infection, Vaccine 35:557–563, 2017.
66
The Use of Prosthetic and Orthotic
Limbs in Avian Medicine
LAURA M. KLEINSCHMIDT
Introduction to Prosthetics and Orthotics (Haliaeëtus leucocephalus), a mallard duck (Anas platyrhyn-
chos), and a domestic goose (Anser anser) with both implan-
Prosthetics and orthotics are a continually evolving treat- tation devices and external prostheses described.5,12,13,18–21
ment modality in veterinary practice, and have been devel- The long-term success rate was variable in these cases,
oping in human medical practice for centuries.1 Prosthetics though most of these patients showed significant improve-
replace a missing limb or body part while orthotics support ment immediately following treatment even if long-term
or protect an injured limb. Two major categorizations of prognosis was poor.
prosthetic devices include internal coaptation (direct skel- In clinical practice, limb prostheses are not routinely
etal attachment, surgical implants) and external coaptation considered a good option for treatment of avian species
devices. Orthotic devices used in veterinary medicine consist due to a higher likelihood of patient refusal and guarded
of external coaptation devices. The devices themselves are long-term prognosis. In waterfowl, raptors, long-legged,
called a “prosthesis” or “prosthetic device” (prosthetics) and other large-bodied birds, solely the need for surgi-
or an “orthosis” or “orthotic device” (orthotics). Use of cal amputation of the legs results in euthanasia in most
prosthetics and orthotics in veterinary medicine has been cases. However, limb prostheses provide an alternative to
reported in the literature for dogs, cattle, goats, horses, euthanasia that would improve animal welfare for select
tortoises, birds, and micropigs.2–21 Many different types of individuals under human care. This also has implications
devices have been used including titanium bone implants, for the conservation of highly endangered species that are
joint replacement implants, braces, boots, slings, carts, and genetically valuable and may otherwise be removed from
even devices made with three-dimensional printers. With the breeding population without alternative options for
improvements in the quality of care provided for veterinary limb replacement.
patients, prosthetics and orthotics are becoming a viable
treatment option for these animals.
The use of prosthetics in avian medicine reported in Choosing the Ideal Avian Candidate
the literature is more limited than in mammalian counter- and Device
parts. Several case reports describe beak prostheses.4,9,10,15
These cases primarily involved a single psittacine (Moluc- In considering the use of these devices, clinicians must weigh
can cockatoo [Cacatua moluccensis]) describing maxillary the pros and cons in each individual patient. Candidates for
(rhamphotheca) fracture and prosthetic placement, and a prosthetic or orthotic device include birds with an injury to
long-beaked birds including a black-neck aracari (Ptero- the legs resulting in neurologic damage, fracture malunion,
glossus aracaris), a channel-billed toucan (Ramphastos vitel- loss of appendages, or surgical amputation with resultant
linus), an African ground hornbill (Bucorvus abyssinicus), inability to ambulate normally. The best candidates for
and a Marabou stork (Leptoptilos crumeniferus), with both orthotic or prosthetic devices are large-bodied or long-legged
mandibular and maxillary devices described.4,9,10,15 Overall, birds that require two functional limbs for effective mobility
beak prostheses were successful in addressing the injuries, and normal day-to-day function. These birds include birds
even if some were removed later in life. Sporadic reports of of prey, waterfowl, galliformes, storks, ibises, herons, cranes,
limb prostheses also exist in the literature and conference flamingoes, and penguins. Most smaller birds (<200 g),
proceedings, including several crane species (a white-naped including most passerine species, are not good candidates
crane [Grus vipio], a Siberian crane [Grus leucogeranus] and for prosthetic or orthotic devices due to their low body
a whooping crane [Grus Americana]), a buzzard (Buteo weights and very small limbs. Although prostheses/orthoses
buteo), a Secretary bird (Sagittarius serpentarius), a bald eagle may be made with lightweight materials, construction of a
465
466 SE C T I O N 12 Avian
prosthetic/orthotic device to accommodate for such a low • BOX 66.1 Example of Intensive Physical
body weight and still allow a small bird to move freely Therapy Schedule
presents significant challenges. These challenges may be
overcome as new technologies become available. Further- • Weeks 1–2: Allow the patient to wear the prosthetic device
for 30 minutes up to four sessions per day, with at least a
more, small bird species including passerines and psittacines 30-minute break between sessions. Every other day, if the
may often compensate with only one functional leg and bird is tolerating four sessions, add one additional session.
may learn to utilize their remaining limb and their beak Continue increasing the number of sessions every other day
to move around their enclosures effectively, making limb up to eight sessions a day.
amputation without prosthetic device placement a more • Weeks 3–4: If the patient has well-tolerated wearing of the
prosthetic device for 30 minutes for eight sessions per day,
viable treatment option in these species. Birds may also move on to this step. Allow the patient to wear the prosthetic
survive adequately without a supportive device if they rely device for 1-hour sessions, with at least a 30-minute break
more heavily on flight as a means of locomotion, although following, starting with up to four sessions per day. If the bird
they may experience some difficulty during take-off or tolerates the four sessions of 1 hour each, every other day
landing. Birds housed outdoors will be at a higher risk for add one additional session. Work up to the patient wearing
the prosthetic for eight sessions of 1 hour each with at least
predation. 30 minutes between sessions per day.
An animal with inherent genetic value, such as endan- • Weeks 5–6: If the patient has accepted the device for
gered species, may be a more desirable candidate in the 1-hour sessions, increase to 2-hour sessions with at least a
captive breeding setting when considering the time and 30-minute break in between. Start with two sessions per day
resources needed for success. Companion birds carry sub- and gradually increase every other day to up to four sessions
per day.
stantial emotional value with their bonded owners and may • Weeks 7–8: If the patient accepted the device for 2-hour
be good candidates as long as the owner is well informed sessions, progress to 4-hour sessions with at least a 1-hour
upon deciding to pursue this treatment option. When break in between sessions. Start with one session per day.
choosing a candidate, the bird’s temperament must come If the bird has tolerated the 4-hour session once per day for
into consideration. Most prosthetic or orthotic devices 1 week, increase to two sessions per day for week 8 with at
least a 1-hour break between.
will not be accepted by an individual or species that is • Progressing from Week 8: Once the animal has accepted the
prone to stressful behavioral responses, and complications device for longer periods without secondary complications,
may include myopathy, immunocompromise secondary to consider increasing the time to 8-hour sessions per day (or
stress, or death. An animal accustomed to human contact more per patient needs).
(educational ambassador, hand-reared, well trained, com-
panion animal, etc.) would make an ideal candidate, as
frequent patient handling may be required. Some bird
species (i.e., psittacines) are more apt to pick at external • BOX 66.2 Examples of Supplemental Physical
coaptation devices. Psittacines may also be less likely to Therapy Modalities
tolerate anything attached to their body externally and may
• Hydrotherapy (swimming)
refuse to move until the offending item is removed, making • Range-of-motion exercises
these animals less ideal candidates. Though any bird may • Massage
react negatively to a novel external device, many birds will • Acupuncture/electrotherapy
adapt over time with the appropriate training and physical • Laser treatment (i.e., affected joints)
• Thermotherapy (heat/cold packing)
therapy. As with any medical treatment, the clinician must
• Balance, coordination, or strengthening exercises
decide if this treatment would be appropriate for each
individual patient. It is recommended to consult with a certified veterinary physical therapist for
further information on these modalities.
Maintaining prosthetic and orthotic devices in animal
patients does require significant dedication and attention to
detail from staff or owners wishing to utilize these devices.
The immediate post-placement period requires intensive (Box 66.2).22 Caretakers must monitor patients closely for
physical therapy in most cases (e.g., physical therapy pro- subtle signs of discomfort or gross lesions indicative of skin
tocol provided in Box 66.1), with gradual increases in time inflammation, infection, ulceration, or pressure sores under
spent with the patient wearing the device until the patient the device due to excessive use, inappropriate fit, or poor
will tolerate the device for the desired length of time. Some hygiene. The contralateral limb must be routinely moni-
birds may not tolerate being handled multiple times a day, tored for muscle damage/breakdown or pododermatitis.
and thus sessions in the device may occur less frequently The device must be cleaned regularly to avoid dermatitis
but still follow a protocol of gradually increasing durations secondary to poor hygiene. The device must be serviced
wearing the device. Each case is different and physical regularly to avoid device failure due to poor maintenance
therapy must be tailored to each individual to accommodate or lack of replacement after excessive wear. Devices may
progress made and the resources/staff available. Additional also be modified over time if the patient’s needs change
physical therapy modalities like those used in companion or be replaced if newer, more innovative devices become
mammal medicine may have applications in avian cases available. With a dedicated and attentive caretaker, birds
CHAPTER 66 The Use of Prosthetic and Orthotic Limbs in Avian Medicine 467
with prosthetic/orthotic devices may do very well and have with injuries that necessitate prosthetic devices often have
an excellent quality of life. additional injuries or ailments, such as myopathy or shock,
Lastly, when choosing a prosthetic/orthotic device for a which require treatment prior to proceeding. Anesthesia,
patient, a clinician must determine the ultimate goals and sedation, or prolonged manual restraint may be required
realistic expectations for any device or individual. By com- during the fabrication process; thus, only a stable patient
bining the bird’s clinical condition and determination of should undergo these procedures. Animals with unrelated
what ideal daily use entails, the clinician may better decide advanced disease processes are likely not ideal candidates for
which type of device is appropriate for any given patient. use of prosthetic devices in general due to their potential
For example, a bird that is routinely handled may do well co-morbidities or imminent mortality.
with an external coaptation device that is easily taken on Once a patient is stabilized, clinicians are encouraged
and off. However, a different patient that requires less fre- to collaborate with veterinary orthopedic surgeons to plan
quent handling, such as an exhibit or breeding animal, may and place internal coaptation prosthetic devices, such
do better with a permanent internal coaptation device or as titanium bone implants or joint replacement devices.
more robust external coaptation device. The decision to try Licensed prosthetists and orthotists (veterinary or human
a prosthetic device over humanely euthanizing an injured medical professionals) are an invaluable resource to develop-
bird may depend on the long-term plans for that individual. ing an innovative solution for each individual patient and
Is the animal a crucial contributor to the population? Will should be consulted from inception to implementation of
the animal be able to successfully breed with the use of an external coaptation device. The easiest way to find a
the supportive device if that is the goal for the individual? prosthetist/orthotist nearby is to consult with a veterinary
How will the device affect this individual’s quality of life? specialty referral or university-based orthopedic surgery
How much additional longevity after device placement is a practice who are most likely to have collaborated with these
reasonable expectation of success in exchange for the time consultants on companion mammal cases; these consultants
and resources devoted to this individual? Unfortunately, would also likely be more willing to help with an avian case
the data available thus far on long-term prognosis with than a prosthetist/orthotist who has never before worked
prosthetic devices in birds are variable by individual and with veterinary patients. However, local human hospitals,
species; there are not enough data with which to make orthopedic surgery, or physical therapy practices may also
definitive conclusions. It is only with further collaboration be able to direct veterinary clinicians to an available licensed
and discussion between colleagues regarding case successes prosthetist/orthotist in the surrounding area. Consultants
and failures that veterinarians will be able to make definitive will likely want to examine the patient and identify the
conclusions about long-term prognosis for any given type specific needs of each individual in order to personalize
of patient or device. the device accordingly; they also have the infrastructure to
create professional-grade devices cost effectively. During the
developmental stage, species-specific and individual needs
Fabrication and Care of a also should be discussed in accordance with previously set
Prosthetic/Orthotic Device expectations for the patient. For example, a waterfowl species
that swims regularly will need a device made of waterproof
Once a clinician has made the decision to try a prosthetic/ materials that allows drainage and dries quickly to prevent
orthotic device, the animal must be prepared accordingly moist dermatitis under the device. Devices should be made
prior to fabrication and placement of the device. Any with a similar color pattern to the natural markings of the
external wounds or surgical incisions should be treated species involved. The weight-bearing surface of the device
in accordance with standard wound care principles and should have adequate traction to prevent patient slippage on
should have healed completely prior to device placement wet or smooth surfaces. A professional with experience and
to avoid irritation, delayed wound healing, or secondary knowledge of material options and device configurations is
infection due to mechanical trauma from the device. The crucial to developing a device with the highest likelihood
placement of a supportive device prior to the resolution of success.
of external wounds could also result in increased pain or The first step in fabrication of a device is to determine the
discomfort during placement or while wearing the device, ideal “normal” for the individual. Preferably with the animal
and therefore could contribute to patient aversion to the awake and standing in normal anatomical position, accurate
device. Bone fractures should also be treated and allowed measurements of the angles of the joints of the limbs should
to heal prior to measuring for the device, as the mechanics be taken of both the normal limb and the affected limb (Fig.
of the limb may change during the orthopedic healing 66.1). Clinicians should utilize the normal contralateral
process. In addition, an external device could alter bone limb as a frame of reference for ideal/normal limb mechanics
healing and result in angular limb deformities or other for the individual. Alternatively, similar-sized conspecifics
complications if utilized prior to complete fracture calcifica- also may be used as a reference of normal anatomy at this
tion. Any other concurrent medical issues should also be stage. Once angle measurements are obtained, the affected
addressed and either resolved or stabilized prior to moving limb must be cast using malleable thermoplastic casting
forward with the fabrication of a prosthetic device. Animals material (Fig. 66.2). A prosthetist/orthotist will then make
468 SE C T I O N 12 Avian
• Figure 66.1 Prior to fabrication of a supportive device, accurate • Figure 66.3 A 1-month-old male domestic goose (Anser anser)
measurements of the angles of the joints of the limbs should be taken presented to a rescue agency missing its right foot. The affected
of both the normal limb and the affected limb. These angles should limb was cast under manual restraint (Fig. 66.2), and a prosthetic
include the angle at which the limb makes contact with the ground device was created using polypropylene plastic that was heated and
and all other acute angles of the joints that will be involved in the vacuum molded to a solid plaster mold made from the hollow original
prosthesis/orthosis. cast. Thermofoam padding and diabetic-grade silicone were used for
internal cushioning, Velcro strapping was used to attach the device to
the patient’s limb, and a nonskid rubber sole was affixed to the distal
end of the device for traction.
A B
• Figure 66.4 (A) Trial and error should be expected to attain the ideal version of a device for each
individual patient. This example shows the first orthotic prototype for a mallard duck (Anas platyrhynchos).
The height of the device was too tall, resulting in the animal leaning heavily toward the unaffected limb.
The angle of the weight-bearing surface was inaccurate, resulting in only the cranial edge of the weight-
bearing surface touching the ground. Finally, the proximal portion of the brace was too long, resulting
in difficult and uncomfortable attachment to the patient due to apposition with the thigh musculature.
Modification of the device was necessary to create a functional orthosis. (B) The final version of the
orthotic device used in this patient following modification for appropriate fit. Note the more normal body
posture and accurate angulation of the weight-bearing surface, resulting in full traction with the ground.
The device was also shortened proximally to prevent apposition with the thigh musculature to prevent
patient discomfort during use.
66.4A, B). Depending on the extent of modifications neces- certain colors or hardware on the device. These modifica-
sary and the patient’s stress level under manual restraint, tions may also be made in the trial and error process to
additional anesthetic or sedation episodes may be needed improve overall patient success.
during the fitting process. The device should fit snugly to Prosthetic and orthotic devices should be maintained
prevent contact sores or skin ulcerations but should not be with proper hygiene and repaired or replaced as needed.
too tight as to cut off circulation to the distal portions of External coaptation devices (see Figs. 66.3 and 66.4)
the leg. The device may need to be modified depending on should be cleaned regularly with a damp cloth and mild
the surrounding soft tissue structures or directionality of antibacterial soap, and then dried with a towel before use.
the feathers to avoid patient discomfort. Additionally, the A less frequent deep cleaning should also be done using a
animal should be observed standing/walking in the device to 50/50 mix of water and rubbing alcohol before allowing
ensure the correct angulation and resultant weight-bearing the device to air-dry prior to use. Extreme temperatures
surface are present to allow the patient to ambulate with a could damage polypropylene plastic prosthetic/orthotic
near-normal gait. The closer to a normal gait, the better, devices and thus should be avoided. Heating elements or
as this will increase normal weight-bearing on the affected devices, such as hair dryers, should not be used to dry
limb and prevent or delay contralateral limb breakdown or these devices to prevent thermal damage. Devices should
pododermatitis. Once an ideal device has been created, the not be left in direct sunlight (e.g., in a hot car) to prevent
animal should start a physical therapy regime as previously their disfiguration. Care instructions will vary depending on
described. Also, the animal should be gradually introduced the building materials used to create the device, and thus
to a variety of substrates, most importantly those present in clinicians should consult with their prosthetist/orthotist on
its natural enclosure. For example, a waterfowl species may specific care instructions for each unique device prior to
have hydrotherapy performed with and without the device use. The need to repair or replace a device over time will
during physical therapy; sessions with the animal wearing vary depending on the degree of use (amount of daily wear
the device should be used to test its compatibility with and tear) and the environment in which the device is used,
submersion and swimming in a controlled setting (i.e., a and thus will be patient dependent. Clinicians may also
large trough filled with water) prior to being introduced to choose to replace devices with innovative models as newer
a natural-type pond in an exhibit or other outdoor setting. technologies become available.
Social species should also experience a gradual return to the
flock to avoid the other animals ostracizing the patient or Conclusion
picking at the device. Depending on the species involved,
attention to detail during the device design process may Prosthetics and orthotics are a viable treatment option
help to prevent these negative social interactions by avoiding for exotic animal practitioners to use to improve quality
470 SE C T I O N 12 Avian
of life and longevity for their animal patients, including 9. Morris PJ, Weigel JP: Methacrylate beak prosthesis in a Marabou
avian patients. As with any treatment option, the pre- stork (Leptoptilos crumeniferus), J Assoc Avian Vet 4.2:103–106,
siding clinician must decide if this treatment would be 1990.
10. Peters JC: Prosthesis of the lower bill of an African ground
appropriate for each individual patient by scrutinizing the
hornbill (Bucorvus abyssinicus), Int Zoo Yearb 3(1):112–113,
species-specific requirements, individual temperament, 1962.
population sustainability, and support staff/caretakers and 11. Prakash KK, Prakash S: Artificial limb for a heifer, Vet Rec
other resources available in each case. The obvious benefits 127(25–26):623, 1990.
of using prosthetics and orthotics include that they are 12. Richter NA, Bacon RD, Richter KJ, et al: Use of a leg prosthe-
an alternative to euthanasia and will allow more normal sis in a bald eagle (Haliaeëtus leucocephalus), J Assoc Avian Vet
weight-bearing and enhanced mobility and thus improve 109–112, 1990.
quality of life, especially in patients who cannot ambu- 13. Rush EM, Turner TM, Montgomery R, et al: Implantation of
late without them (bilateral amputations in quadrupeds, a titanium partial limb prosthesis in a white-naped crane (Grus
large mammals that require distribution of their weight vipio), J Avian Med Surg 26(3):167–175, 2012.
on four limbs to ambulate, or bipedal animals such as 14. Saunders MM, Brecht JS, Verstraete MC, et al: Lower limb direct
skeletal attachment: a Yucatan micropig pilot study, J Invest Surg
birds). Possible risks include patient refusal to use the device
25(6):387–397, 2012.
despite physical therapy—the physical therapy itself being 15. Sleamaker TF: Prosthetic beak for a salmon-crested cockatoo, J
time and labor-intensive—underlying skin inflammation, Am Vet Med Assoc 183(11):1300–1301, 1983.
infection, or ulceration due to poor hygiene or inappropri- 16. St-Jean G: Amputation and prosthesis, Vet Clin North Am Food
ate fit, contralateral limb muscle damage/breakdown or Anim Pract 12(1):249–261, 1996.
pododermatitis, and device upkeep, including regular clean- 17. Zehr DR: Amputation and use of a prosthesis, Mod Vet Pract
ing, maintenance, and replacement of devices over time. 58(11):933–934, 1977.
Clinicians interested in using these devices are encouraged 18. Kleinschmidt LM, Ellsworth V, Hoppes SM: Use of orthotics
to collaborate with other veterinary (or human) medical and prosthetics for distal limb injuries in waterfowl. In Proceed-
professionals, especially licensed prosthetists/orthotists. Any ings of the American Association of Zoo Veterinarians (AAZV)
prosthetic/orthotic device will require a level of innovation Annual Conference 2016, pp 65–66.
19. Kapustin N, Bear-Hull D, Teare JA: Use of a novel prosthesis for
and creativity because each device must be individualized
treatment of lameness in a whooping crane. In Proceedings of
for each patient and a unique situation. Collaboration and the American Association of Zoo Veterinarians (AAZV) Annual
discussion between veterinary professionals of both case Conference 2008, p 156.
successes and failures are crucial to the further development 20. Bennett RA, O’Brien ET: Use of a finger joint prosthesis for the
of prosthetic and orthotic device use in avian patients. management of tarsal arthritis in a Siberian crane. In Proceed-
ings of the American Association of Zoo Veterinarians (AAZV)
References Annual Conference 1999, pp 279–282.
21. McManamon R, Reynolds JR, Greenwood KM: Application of
1. Stryła W, Pogorzała AM, Kasior I, et al: Limb amputations a leg prosthesis on a secretary bird (Sagittarius serpentarius). In
from the ancient times to the present, Pol Orthop Traumatol Proceedings of the American Association of Zoo Veterinarians
78:155–166, 2013. (AAZV) Annual Conference 1992, p 180.
2. Adamson C, Kaufmann M, Levine D, et al: Assistive devices, 22. Rivera PL: Canine rehabilitation therapies I and II. Proceedings
orthotics, and prosthetics, Vet Clin North Am Small Anim Pract of the 79th Annual Western Veterinary Conference 2007, p 11.
35(6):1441–1451, 2005. 23. Hawkins MG, Barron HW, Speer BL: Birds: chemical restraint/
3. Clabaugh K, Haag KM, Hanley CS, et al: Undifferentiated anesthetic/analgesic agents used in birds. In Carpenter JW,
sarcoma resolved by forelimb amputation and prosthesis in Marion CJ, editors: Exotic animal formulary, ed 4, St. Louis,
a radiated tortoise (Geochelone radiata), J Zoo Wildl Med 2013, Elsevier Saunders.
36(1):117–120, 2005. 24. Heard D: Anesthesia. In Speer B, editor: Current therapy in avian
4. Crosta L: Alloplastic and heteroplasic bill prostheses in 2 ram- medicine and surgery, ed 1, St. Louis, 2016, Elsevier.
phastidae birds, J Avian Med Surg 16(3):218–222, 2001. 25. Marx KL: Therapeutic agents. In Harrison GJ, Lightfoot TL,
5. David T: Knee-joint prosthesis in a buzzard (Buteo Buteo), editors: Clinical avian medicine (vol 1). Palm Beach, FL, 2006,
Tierarztl Prax 4(3):387–389, 1976. Spix Publishing.
6. Desrochers A, St-Jean G, Anderson DE: Limb amputation and 26. Zehnder AM, Hawkins MG, Pascoe PJ, et al: Bird anesthesia. In
prosthesis, Vet Clin North Am Food Anim Pract 30(1):143–155, West G, Heard D, Caulkett N, editors: Zoo animal and wildlife
2014. immobilization and anesthesia, ed 2, Ames, IA, 2014, John Wiley
7. Hall CW: Developing a permanently attached artificial limb, Bull & Sons, Inc.
Prosthet Res:144–157, 1974.
8. Koger LM, McIlhattan J, Schladetzky R: Prosthesis for
partially amputated foreleg in a horse, J Am Vet Med Assoc
156(11):1600–1604, 1970.
67
Avian Spirurids
CARLES JUAN-SALLÉS AND MICH AEL M. GARNER
471
TABLE
67.1 Main Pathogenic Spirurids Described in Wild and Captive Birds, With Avian and Intermediate Hosts, and Organ Targets
472 SE C T I O N 12 Avian
Family (Superfamily) Species Avian Hosts Intermediate Hosts Organ Targets References
Acuariidae (Acuaroidea) Dispharynx spp. (particularly Wide range of orders Pillbug, sowbug Proventriculus, esophagus 7, 22, 26, 39,
D. nasuta) 42, 43
Acuaria skrjabini Finches Unknown Ventriculus 25, 34
Cheilospirura hamulosa Game gallinaceous birds, pheasants Grasshopper, beetle, Ventriculus 14
weevil, sand hopper
Echinuria uncinata Ducks, geese, swans Water fleas Proventriculus, intestine, body 52
walls
Streptocara crassicauda, S. Wild ducks; geese Amphipod crustaceans Ventriculus, esophagus, pharynx Reviewed in 1
incognita (Gammarus spp.,
Hyalella azteca)
S. incognita Chilean flamingos (Phoenicopterus Proventriculus, ventriculus, Reviewed in 1
chilensis) esophagus
S. incognita Mute swans (Cygnus olor) Oropharynx, esophagus 1
Tetrameridae Tetrameres spp. (wide range Wide range of species, usually Mostly coleopteran and Proventriculus 27, 35, 46
(Habronematoidea) of species, disputed) aquatic orthopteran insects,
also crustaceans
Microtetrameres spp. (wide Wide range of species, usually At least orthopteran Proventriculus 19, 27
range of species) terrestrial insects
Geopetitia aspiculata Passerine birds, Charadriiformes, Crickets and Distal serosal surface of 27
Coraciiformes cockroaches esophagus, proventriculus
and ventriculus
Habronematidae Procyrnea sp. Passerine birds; black-backed Insect suspected but Ventriculus; also proventricular 38, 45
(Habronematoidea) woodpecker (Picoides arcticus) not identified and intestinal lumen
Hadjelia truncata Domestic pigeons; also Beetles Ventriculus 36, 44
Coraciiformes, helmeted
guineafowl (Numida meleagris)
Sicarius uncinipenis Greater rhea (Rhea americana) Unidentified arthropods Ventriculus 9, 53
Onchocercidae Sarconema eurycerca Swans and geese (Branta, Anser) Biting louse Heart 33, 51
(Filaroidea)
Pelecitus sp., P. tercostatus, Psittacine birds, channel-billed Lice, mosquitoes, other Subcutaneous tissue around 2, 10, 32
P. calamiformis toucan (Ramphastos vitellinus) arthropods joints (especially hock)
Chandlerella quiscali Emus (Dromaius novaehollandiae), Midges (Culicoides spp.) Brain, spinal cord 16, 31
northern crested caracara
(Caracara cheriway)
Paronchocerca ciconarum Storks Unknown Heart, pulmonary vessels 15
P. tonkinensis Rufus-crested bustards (Eupoditis Unknown Pulmonary arteries (adults); 37
ruficrista) microfilariae in tissues
Diplotriaenidae Serratospiculum spp. Falcons; also other raptors and Beetles, grasshoppers, Air sacs 23, 40, 41, 49
(Diplotriaenoidea) passerine birds locusts, wood lice,
crickets
Serratospiculoides Falco spp., Cooper’s hawk (Accipiter Grasshoppers, crickets Air sacs 24, 28, 47
amaculata cooperii), great tit (Parus major)
Monopetalonema alcedinis Belted kingfisher (Megaceryle alcyon) Unidentified insects Air sacs 6
and other kingfishers
Thelaziidae Oxyspirura mansoni Wide range of orders Surinam cockroach Conjunctival sac 50
(Thelazioidea)
O. petrowi Over 80 species worldwide Unidentified arthropods Eyelids, nictitating membrane, 11, 12
nasolacrimal duct, Harderian
gland, intraorbital tissues
Thelazia anolabiata, Thelazia Numerous species; disease in Dipteran flies Periocular tissues 17
spp. Andean Cock of the Rock
(Rupicola peruviana) and Senegal’s
parrot (Poicephalus senegalus)
Ceratospira inglisi Pigeons, doves, cockatoos Presumably flies Periocular tissues 48
Gongylonematidae Gongylonema sp. Scops owl (Otus scops), presumed Around 50 species of Oral cavity 18
(Spirudoidea) accidental host arthropods (mostly
coprophagus)
Gongylonema sp. Horned owls (Asio otus) Esophagus 30
Gnathostomatidae Gnathostoma spp. Fish-eating birds Copepods first, then fish Skeletal muscles 20
(Gnathostomatoidea) and amphibians; birds
are parathenic hosts
Physalopteridae Physaloptera sp. Bobwhite quail (Colinus virginianus) Orthopterans, beetles Skeletal muscles 5
(Physalopteroidea)
P. alata Eurasian bittern (Botaurus stellaris) Stomach 29
CHAPTER 67 Avian Spirurids
473
TABLE Clinical Findings and Lesions Commonly Reported With the Main Pathogenic Spirurids of Wild and Captive Birds. Treatment Protocols, When
67.2 Proved to Be Effective, Are Also Included
Acuaria skrjabini Lethargy, inappetence, diarrhea, Necrotizing ventriculitis, koilin degeneration and 25, 34
weight loss thickening, secondary bacterial infection;
nematodes burrow in the interface between koilin
and mucosal glands
Cheilospirura hamulosa Anemia, emaciation Ventriculitis with leiomyositis 14
Echinuria uncinata Emaciation, anemia, eosinophilia, Proventricular thickening with excessive mucus; 52
heterophilia granulomas with intralesional nematodes
Streptocara crassicauda, Emaciation, debilitation Ulcerative or necrotizing ventriculitis, esophagitis or Reviewed
S. incognita pharyngitis in 1
S. incognita Found dead (debilitating condition, Ulcerative ventriculitis and proventriculitis with Reviewed
cachexia) intralesional nematodes and blood-filled cystic in 1
lesions in ventricular muscular layers
S. incognita Anorexia, lethargy, reluctance to Fibrinonecrotizing pharyngitis and esophagitis with 1
move intralesional nematodes
Tetrameres spp. (wide Weakness, loss of appetite, diarrhea, Proventricular thickening, compression atrophy of 27, 35, 46
range of species, eosinophilia, anemia; associated glands by female nematodes, necrosis; fistulated
disputed) with mortality in wild cranes proventricular nodules
Microtetrameres spp. Generally not described; anorexia, Glandular compression atrophy, ductal hyperplasia/ 19, 27
(wide range of weight loss, lethargy, and goblet cell metaplasia of proventricular glands,
species) leukocytosis in hornbills mild glandular necrosis, proventriculitis and
hemorrhage; female nematodes hematophagous
Geopetitia aspiculata Weight loss, distension of coelomic Granulomatous proventriculitis and visceral Ivermectin, fenbendazol 27
cavity coelomitis around encysted nematodes; posterior
end of some nematodes in the lumen of
proventriculus
Procyrnea sp. Lethargy, perching low or on ground, Koilin fragmentation and thickening with intralesional 38, 45
emaciation nematodes and secondary bacterial and Candida
infections, ventriculitis, ulceration, hemorrhage;
associated with amyloidosis in passerine birds
Hadjelia truncata Weight loss, poor food consumption, Ventricular enlargement, koilin disruption/ 15 mL of levamisole solution (37.5 mg 36, 44
diarrhea, weakness, reduced degeneration with clear spaces, abundant levamisole hydrochloride/mL) in 1 L
packed cell volume (PCV) and nematodes between koilin and mucosa, of drinking water36
total proteins, mortality (pigeons) secondary bacterial colonization (pigeons)
Sicarius uncinipenis Weight loss, hyporexia/anorexia Koilin ulceration/degeneration, ventriculitis, red 9, 53
nematodes embedded in the mucosa
Sarconema eurycerca Emaciation, cardiac failure Myocarditis with hemorrhagic tracts, vasculitis/ 33, 51
fibrinoid necrosis, mineralization, adult nematodes
and microfilariae
Pelecitus sp., P. Lameness, nodular thickening of soft Nodular thickening of soft tissues around joint, 2, 10, 32
tercostatus, P. tissues around joint fibrinous tenosynovitis
calamiformis
Chandlerella quiscali Torticollis, progressive ataxia Encephalitis 31
Paronchocerca Myocardial degeneration, thrombosis and thickening 15
ciconarum of pulmonary arteries
P. tonkinensis Pneumonia; association with cardiac rupture, 37
myocardial degeneration and fibrosis
Serratospiculum spp. Dyspnea, reduced fitness in flight, Air sacculitis, pulmonary necrosis, Melarsomine (0.25 mg/kg IM for 2 23, 40, 41,
weight loss, lethargy; also bronchopneumonia, emaciation; bone fractures days) followed 10 days later by a 49
vomiting, bone fractures noted in all infected wild raptors in one single dose of ivermectin (1 mg/kg
retrospective study in Italy IM) for S. seurati49
Serratospiculoides Subclinical if parasites not abundant, Air sacculitis with fibrosis, pneumonia; 24, 28, 47
amaculata but can cause signs similar meningomyelitis with spongiosis and axonal
to Serratospiculum; anorexia, degeneration in a falcon with atypical parasite
recumbent and hindlimb deficits migration into vertebral canal
(falcon with atypical parasite
migration into vertebral canal)
Monopetalonema Not available Air sacculitis, pneumonia 6
alcedinis
Oxyspirura mansoni Conjunctivitis, association with Conjunctivitis, bilateral cataracts (histopathological Removal of nematodes, topical 50
blindness and cataracts findings not provided) in zoo pheasants 0.5% iodine or 0.5% Lysol, 0.5%
diethylcarbamazine in drinking water
O. petrowi Ocular inflammation, corneal opacity Harderian adenitis with ductal dilatation, fibrosis and 11, 12
gland atrophy; corneal ulceration and opacity
Thelazia anolabiata, Keratoconjunctivitis Keratoconjunctivitis, hyphema, blepharospasm Removal of nematodes, ivermectin 17
Thelazia spp. 0.2 mg/kg IM single dose,
topical ciprofloxacin and epithelial
regenerator
Ceratospira inglisi Chemosis Chemosis, subclinical Removal of nematodes, topical 48
Ivomec 1% (1 drop), triple antibiotic
ophthalmic ointment
Gongylonema sp. Starvation, emaciation, plaques in Necrotic stomatitis Fenbendazol 50 mg/kg PO, daily 18
the oral mucosae debridement, and supportive therapy
Gongylonema sp. Weakness Obstruction of esophagus with masses of Fenbendazole 33 mg/kg PO for 3 30
nematodes attached to the mucosa consecutive days
Gnathostoma spp. Not reported Rhabdomyositis with fibrosis 20
Physaloptera sp. Not reported Necrotizing and granulomatous rhabomyositis with 5
CHAPTER 67 Avian Spirurids
A
• Figure 67.1 Note cuticular cordons (arrow) in the anterior end of a
specimen of Dispharynx obtained from a parrot (Barnardius zonarius
semitorquatus) with adenomatous proliferative proventriculitis. (Photo
courtesy Dr. Nuhacet Fernández [Loro Parque, Spain].)
B
• Figure 67.4 (A) Gross appearance of low numbers of female
• Figure 67.2 Photomicrograph of a polypoid proventricular lesion
Tetrameres as raised reddish foci scattered in the proventricular
(long arrow) around a female acuariid spirurid (black arrowheads) mucosa of a rock pigeon. Note also an unidentified nematode attached
attached to the mucosa in a musk lorikeet (Glossopsitta concinna). to the mucosa (arrow). (Photo Courtesy Dr. Andrés Montesinos Barceló
Note abundant eggs in the uterus (short arrow). Fibrosis (f) and [Centro Veterinario Los Sauces, Spain].) (B) Photomicrograph of the
inflammation (red arrowheads) are prominent. Inset: detail of cuticular proventriculus depicted in Fig. 67.4A, in which proventricular glands
cordons (arrowheads). m, Muscular tunics. Hematoxylin and eosin. are dilated due to the presence of tetramerid, globoid nematodes with
abundant eggs, eosinophilic fluid (arrows) in the pseudocoelom, and
heavily pigmented intestinal brush border (black arrowheads). Glands
are severely atrophied (red arrowheads) due to compression by the
large, gravid Tetrameres females. Inset: higher magnification of char-
acteristic oval, thick-shelled embryonated spirurid eggs. Hematoxylin
and eosin.
A B
• Figure 67.7 (A) Photomicrograph of the lung of a bearded barbet (Lybius dubius) with severe para-
bronchial hemorrhage (asterisks) associated with microfilariasis. Note foci of pneumonia and atelectasis
(arrowheads). Hematoxylin and eosin. (B) Higher magnification of a pneumonic focus in Fig. 67.6A. Areas
of perivascular (arrowheads) and interstitial inflammation with collapse of air capillaries (atelectasis) and
hemorrhage contain intralesional microfilariae (arrows). Hematoxylin and eosin.
References
• Figure 67.8 Gross image of serratospiculosis in a falcon from 1. Alić A, Prasović S, Hodzić A, et al: Fatal verminous pharyngitis
Colombia. Several adult nematodes are present in the lumen of the and esophagitis caused by Streptocara incognita in mute swans
right caudal pulmonary air sac; parasitism is associated with moderate (Cygnus olor), Avian Dis 57:147–151, 2013.
air sac opacity and fibrin deposit (arrow). (Photo courtesy Dr. Delio 2. Anderson RC: Nematode parasites of vertebrates: Their development
Orjuela [Fundación Paz Animal, Colombia].) and transmission, Oxon, 2000, CAB International.
3. Bartlett CM: Filaroid nematodes. In Atkinson CT, Thomas NJ,
Hunter DB, editors: Parasitic diseases of wild birds, Ames, IA,
Middle East.40,41,49 In North America, serratospiculosis has 2008, Wiley-Blackwell, pp 439–462.
been reported to occur with Serratospiculoides amaculata.24,47 4. Bleier T, Hetzel U, Bauer C, et al: Gongylonema pulchrum infec-
While some studies question the clinical impact of serrato- tion and esophageal squamous cell carcinoma in a vari (Lemur
spiculosis, others point to relevant disease or lesions.41,49 A macaco variegata; Kehr 1792), J Zoo Wildl Med 36:342–345,
recent study of serratospiculosis caused by S. seurati revealed 2005.
an incidence close to 9% in over 1700 falcons, with over 5. Boggs JF, Peoples AD, Lochmiller RL, et al: Occurrence and
70% presenting primarily with dyspnea and reduced fitness pathology of physalopterid larvae infections in bobwhite quail
on flight; anorexia, lethargy, and vomiting can also occur.49 from western Oklahoma, Proc Okla Acad Sci 70:29–31, 1990.
Affected wild raptors frequently are found with fractures, and 6. Bronson E, Kelly K, Hoberg EP: Air sac nematode Monopeta-
it has been suggested that serratospiculosis may predispose lonema alcedinis in a belted kingfisher (Megaceryle alcyon) in
Maryland, USA, J Wildl Dis 50:938–941, 2014.
to trauma and decreased predatory capacity.41,47 Lesions of
7. Churria CD, Spinsanti E, Origlia J, et al: Dispharynx nasuta
serratospiculosis consist of air sacculitis (see Fig. 67.8) with (Nematoda:Acuariidae) infection causing proventricular lesions
fibrosis as well as air sac muscular, epithelial, and mesothelial and death in three captive rosellas (Psittaciformes:Psittacidae), J
hyperplasia associated with nematodes and their eggs, which Zoo Wildl Med 42:164–165, 2011.
may also be present in the lungs and cause pneumonia 8. Cosgrove GE, Lushbaugh WB, Humason G, et al: Synhimantus
with necrotic foci.41,47 A combination of melarsomine and (nematode) associated with gastric squamous tumors in muskrats,
ivermectin has proved effective in resolving symptoms with Wildl Dis 4:54–57, 1968.
CHAPTER 67 Avian Spirurids 479
9. de Oliveira Avelar I, Ribeiro de Almeida L, Andrade dos Santos the American Association of Zoo Veterinarians/European Asso-
H, et al: Sicarius uncinipenis and Deletrocephalus cesarpintoi in ciation of Zoo Wildlife Veterinarians/Institute of Zoo Wildlife
captive greater rheas of Minas Gerais State, Brazil, Braz J Vet Conference 2016, pp 62–63.
Parasitol 23:355–359, 2014. 27. Kinsella JM, Forrester DJ: Tetrameridosis. In Atkinson CT,
10. Díaz JI, Di Nucci DL, Falzone MP, et al: Pelecitus tercostatus Thomas NJ, Hunter DB, editors: Parasitic diseases of wild birds,
(Molin, 1960) (Nematoda, Onchocercidae) in Amazona vinacea Ames, IA, 2008, Wiley-Blackwell, pp 376–383.
(Aves, Psittaciformes) from Argentina: morphological details and 28. Königová A, Molnár L, Hrčková G, et al: The first report of
clinical findings, Acta Parasitol 57:194–198, 2012. serratospiculiasis in great tit (Parus major) in Slovakia, Helmin-
11. Dunham NR, Peper ST, Baxter CE, et al: The parasitic eyeworm thologia 50:254–260, 2013.
Oxyspirura petrowi as a possible cause of decline in the threatened 29. Kordafshari S, Samani R, Hosseini SH, et al: Case report on
lesser prairie-chicken (Tympanuchus pallidicinctus), PLoS ONE Physaloptera alata infection in Botarous stellaris in Iran, World J
9:e108244, 2014. Zool 5:246–248, 2010.
12. Dunham NR, Reed S, Rollins D, et al: Oxyspirura petrowi infec- 30. Kummerfeld N, Stoye M: Uber nachweis und therapie von
tion leads to pathological consequences in Northern bobwhite Gongylonema sp. bei waldohreulen (Asio otus), Praktische Tierarzt
(Colinus virginianus), Int J Parasitol Parasites Wildl 5:273–276, 61:956–960, 1980.
2016. 31. Law JM, Tully TN, Stewart TB: Verminous encephalitis appar-
13. Dvir E, Clift SJ: Evaluation of selected growth factor expression ently caused by the filarioid nematode Chandlerella quiscali in
in canine spirocercosis (Spirocerca lupi)-associated non-neoplastic emus (Dromaius novaehollandiae), Avian Dis 37:597–601, 1993.
nodules and sarcomas, Vet Parasitol 174:257–266, 2010. 32. Madani SA, Dorrestein GM: Filarial tenosynovitis caused by
14. Ebrahimi M, Rouhani S, Mobedi I, et al: Prevalence and Pelecitus species (Spirurida, Filarioidea, Onchocercidae) in the
morphological characterization of Cheilospirura hamulosa, legs of a channel-billed toucan (Ramphastos vitellinus), J Avian
Diesing, 1861 (Nematoda: Acuarioidea), from partridges in Med Surg 26:36–39, 2012.
Iran, J Parasitol Res 2015; Article ID 569340, 6 pages. http:// 33. Manno C, Blake D, Ghisleni G, et al: Cardiac filariosis in migra-
dx.doi.org/10.1155/2015/569340. tory mute swans (Cygnus olor) in Sicily, Int J Health Anim Sci Food
15. Echols MS, Craig TM, Speer BL: Heartworm (Paronchocerca Saf 3:20–27, 2016.
ciconarum) infection in 2 saddle-billed storks (Ephippiorhynchus 34. McOrist S, Barton NJ, Black DG: Acuaris skrjabini infection of
senegalensis), J Avian Med Surg 14:42–47, 2000. the gizzard of finches, Avian Dis 26:957–960, 1982.
16. Edwards EE, Dangoudoubiyam S, Hoppes SM, et al: Granulo- 35. Mowlavi GR, Massoud J, Mobedi I, et al: Tetrameres (Tetrameres
matous filarial encephalomyelitis caused by Chandlerella quiscali grusi) (Shumakovich, 1946) (Nematoda: Tetrameridae) in Eur-
in a northern crested caracara (Caracara cheriway), J Zoo Wildl asian cranes (Grus grus) in central Iran, J Wildl Dis 42:397–401,
Med 48:237–240, 2017. 2006.
17. Elías R, Mamani J, Hermoza C, et al: First report of thelaziosis 36. Navabi R, Khedri J, Jahantigh M: High clinical disturbance and
(Thelazia anolabiata) in an Andean Cock of the Rock (Rupicola mortality in pigeon flocks caused by Hadjelia truncata infection
peruviana) from Peru, Vet Parasitol 158:382–383, 2008. in Sistan, Iran, Turk J Vet Anim Sci 37:355–357, 2013.
18. Esperón F, Martín MP, Lopes F, et al: Gongylonema sp. infection 37. Nicholls PK, Bailey TA, Gibbons LM, et al: Parasitic infec-
in the scops owl (Otus scops), Parasitol Int 62:502–504, 2013. tion in a flock of rufous-crested bustards (Eupodotis ruficrista)
19. Ferrell ST, Pope KA, Gardiner C, et al: Proventricular nemato- in the United Arab Emirates, J Zoo Wildl Med 26:590–596,
diasis in wrinkled hornbills (Aceros corrugatus), J Zoo Wildl Med 1995.
40:543–550, 2009. 38. Niemuth JN, Allgood JV, Flowers JR, et al: Ventricular habrone-
20. García-Márquez LJ, León-Règagnon V, Lamothe-Argumedo miasis in aviary passerines, Case Rep Vet Med 2013:2013, article
R, et al: Inflammatory response caused by larvae and adults ID 719465, 6 pages. doi:10.1155/2013/719465.
of Gnathostoma (Nematoda: Gnathostomatidae) in vertebrates 39. Raymond JT, Miller CL, Garner MM: Adenomatous prolifera-
of Mexico, including humans, Rev Mex Biodiv 85:429–435, tive proventriculitis (APP) in captive birds. In Proceedings of the
2014. American Association of Zoo Veterinarians 2002, pp 94–95.
21. Gardiner CH, Poynton SL: An atlas of metazoan parasites in 40. Samour JH, Naldo J: Serratospiculiasis in captive falcons in the
animal tissues, Washington, DC, 1999, Armed Forces Institute Middle East: a review, J Avian Med Surg 15:2–9, 2001.
of Pathology. 41. Santoro M, D’Alessio N, Di Prisco F, et al: The occurrence
22. González-Hein G, Fredes F, Kinsella M, et al: New reports of and pathogenicity of Serratospiculum tendo (Nematoda: Diplo-
helminthes in captive exotic psittacine birds in Chile, Arch Med triaenoidea) in birds of prey from southern Italy, J Helminthol
Vet 44:87–91, 2012. 90:294–297, 2016.
23. Green CH, Gartrell BD, Charleston WA: Serratospiculosis 42. Santoro M, Tripepi M, Kinsella JM, et al: Helminth infestation
in a New Zealand falcon (Falco novaeseelandiae), N Z Vet J in birds of prey (Accipitriformes and Falconiformes) in Southern
54:198–201, 2006. Italy, Vet J 186:119–122, 2010.
24. Hawkins MG, Couto S, Tell LA, et al: Atypical parasitic migration 43. Schulman FY, Montali RJ, Citino SB: Pathology, diagnosis, and
and necrotizing sacral myelitis due to Serratospiculoides amaculata treatment of Synhimantus nasuta infection in African jacanas
in a prairie falcon (Falco mexicanus), Avian Dis 45:276–283, (Actophilornis africana), J Zoo Wildl Med 23:313–317, 1992.
2001. 44. Sentíes-Cué CG, Charlton BR, Anthenill L, et al: Parasitic ven-
25. Hindmarsh M, Ward K: Mortality of finches (family Estrildidae) triculitis caused by Hadjelia truncata in California rock pigeons
caused by (Acuaria skrjabini), Aust Vet J 70:451–452, 1993. (Columba livia), J Vet Diagn Invest 23:1243–1246, 2011.
26. Juan-Sallés C, Garner MM, Fernández N, et al: Retrospective 45. Siegel RB, Bond ML, Wilkerson RL, et al: Lethal Procyrnea
study of proventricular acuariid spiruridiasis in captive psittacine infection in a black-backed woodpecker (Picoides arcticus) from
birds with proliferative proventricular disease. In Proceedings of California, J Zoo Wildl Med 43:421–424, 2012.
480 SE C T I O N 12 Avian
46. Silva RJ, Oliveira-Sequeira TCG, Gurgel CC: Occurrence of 50. Vellayan S, Jeffery J, Oothuman P, et al: Oxyspiruriasis in zoo
Tetrameres confusa (Nematoda, Tetrameridae) in Ara ararauna birds, Trop Biomed 29:304–307, 2012.
(Psittacidae), Arq Bras Med Vet Zootec 57:562–564, 2005. 51. Woo GH, Jean YH, Bak EJ, et al: Myocarditis by nematodes
47. Sterner MC, Cole CA: Diplotriaena, Serratospiculum and infection, presumably Sarconema eurycerca, in a wild whooper
Serratospiculoides. In Atkinson CT, Thomas NJ, Hunter DB, swan (Cygnus cygnus) in Korea, J Vet Med Sci 72:1233–1235,
editors: Parasitic diseases of wild birds, Ames, IA, 2008, Wiley- 2010.
Blackwell, pp 434–438. 52. Work TM, Meteyer CU, Cole RA: Mortality in Laysan ducks
48. Suedmeyer WK, Smith T, Moore C, et al: Ceratospira inglisi (Anas laysanensis) by emaciation complicated by Echinuria unci-
ocular infestation in a Wompoo fruit-dove (Ptilinopus magnifi- nata on Laysan Island, Hawaii, 1993, J Wildl Dis 40:110–114,
cus), J Avian Med Surg 13:261–264, 1999. 2004.
49. Tarello W: Serratospiculosis in falcons from Kuwait: incidence, 53. Zettermann CD, Nascimento AA, Tebaldi JA, et al: Observa-
pathogenicity and treatment with melarsomine and ivermectin, tions on helminth infections of free-living and captive rheas
Parasite 13:59–63, 2006. (Rhea americana) in Brazil, Vet Parasitol 129:169–172, 2005.
68
Selected Medical Aspects of Bird
Reproduction in Ex Situ Conservation
DANTE LUIS DI NUCCI
T
he knowledge of aviculture techniques has had and physical examinations, hematology, biochemistry, and other
continues to have a significant role in the pet bird necessary diagnostic testing such as imaging techniques
industry, commercial poultry farming, zoos, and (radiography, laparoscopy, or ultrasonography).3,5–8
ex situ bird conservation programs. Captive breeding is Avian reproductive disorders are a result of a complex
already underway or recommended for consideration as a combination of hormonal, physiologic, and behavioral
conservation action for a total of 257 avian species (2.6% actions related to photoperiod, food availability, and suit-
of extant species).1 ability of nest sites, among other factors.9 There are several
Establishing a successful program of captive breeding comprehensive works on the anatomy of the reproduc-
and incubation begins with familiarization with the natural tive tract and reproductive disorders, and their diagnosis,
history and biology of the species, selection of individuals treatment, and prevention observed in Psitaciformes, Gal-
and compatible breeding pairs, knowledge of minimum liniformes, and Anseriformes.3,5,6,9–12 Box 68.1 describes the
needs for captive management (enclosures, enrichment, most common female and male reproductive disorders.
nutrition, health programs), artificial incubation techniques
(equipment, understanding of avian egg and embryo devel- Artificial Incubation
opment, protocols for sanitation, incubation, egg manage-
ment, record keeping, and egg necropsy), and knowledge Artificial incubation should be considered an essential
of pathologies of reproductive organs, which can cause tool for the conservation of rare and endangered species.
infertility, affect the normal development of the embryo, Many bird species will lay replacement eggs if the first
or lead to neonatal diseases. clutch of eggs is removed or lost, creating an opportunity
This chapter summarizes information on medical aspects for increasing reproductive productivity. In some cases a
to aid in the diagnosis and understands possible causes combination of natural and artificial incubation may yield
of infertility, embryonic deaths, and consequent neonatal the best results. In this case, protocols for removal of eggs
complications of captive breeding. for artificial incubation should be carefully planned so as
not to negatively interfere in embryonic development and
Reproduction hatching.13
Those contemplating the implementation of an artificial
Infertility can be defined as a lack of egg production or incubation and hand-rearing program should plan on an
increased frequency of infertile eggs. Although the majority investment in time and economic costs because there are
of infertility problems are due to poor management, diseases demanding tasks in effort, equipment, and building infra-
must be ruled out as well.2 Common causes of infertility structure requirements that must be considered to maximize
include incompatibility of breeding pairs, same-sex pairings the success of the program. Separate rooms or facilities are
(mostly occurring in birds without sexual dimorphism), recommended for storage of eggs, incubation, nursery, and
lack of or difficulty copulating (e.g., underlying disease or grow out rooms. Areas of egg storage should have humid-
physical disorders), breeding immature birds, poor nutri- ity and temperature controlled. Incubation rooms must be
tional status, nutritional deficiencies, obesity, poor body equipped with incubators, hatchers, and egg candlers. The
score, lack of environmental stimuli (e.g., artificial lighting), nursery must be equipped to handle altricial and precocial
stress, inadequate genetic management of the population, species. Animal intensive care units are essential for success-
cloacal abnormalities, and other reproductive disorders.2–4 ful rearing of altricial species. Sufficient space and setting
Diagnosing infertility problems requires an in-depth review according to type of bird to be raised are needed for precocial
of management and breeding records, as well as performing species. A unidirectional work flow throughout the facility
481
482 SE C T I O N 12 Avian
• BOX 68.1 Common Avian There are three critical points during the preincubation
Reproductive Disorders period: collection and handling, storage, and disinfection
of eggs. Careful handling helps prevent injury (shell break-
Reproductive Disorders in Females age) and microbial infection of eggs. Proper collection
Dystocia, egg binding, and egg bound and handling techniques include protection from physical
Chronic egg laying
damage (cracked shells, freezing, or overheating), prevent-
Egg-related coelomitis and coelomitis of reproductive origin
Ectopic ovulation ing damage from sudden or rough movement, and speed
Oophoritis of transfer. For incubated eggs a rapid transfer (less than 5
Cystic hyperplasia of the oviduct and ovary minutes) between nests and/or incubators is recommended.
Retained cystic right oviduct The most frequent pathogens in poultry hatcheries are
Ovarian torsion
Pseudomonas sp., Escherichia coli, Salmonella sp., Mycoplasma
Ovarian and oviductal neoplasia (adenocarcinoma, granulosa-
theca cell tumor, arrhenoma and arrhenoblastoma, ovarian sp., and Aspergillus fumigatus.16 In a study in rheas (Rhea
sertoli cell tumors, dysgerminomas, leiomyoma) americana), 13 bacterial agents (Acinetobacter sp., Aeromo-
Oviductal impaction nas sp., Alcaligenes sp., Bacillus sp., Cedecea sp., Citrobacter
Oviductal prolapse freundii, E. coli, Proteus sp., Pseudomonas sp., Salmonella
Oviductal rupture
gallinarum, Serratia ficaria, Serratia marcescens, Staphylococ-
Oviductal volvulus
Salpingitis, metritis, and oophoritis cus aureus) and four fungal agents (Alternaria sp., Aspergillus
Prolapse and miscellaneous disease of the cloaca sp., Fusarium sp., Mucor sp.) were isolated from eggs with
microbial contamination during the incubation process.17
Reproductive Disorders in Males Options for egg disinfection include dry cleaning,
Orchitis, or epididymitis spraying, and dipping. The dry cleaning method is the
Cystic dilation of the seminiferous tubules and testes
recommended form of disinfection and is preferred because
Testicular neoplasia (sertoli cell tumor, seminomas, mixed germ
cell–sex cord–stromal tumor, arrhenoblastomas, testicular it is safer for the embryos. Dry cleaning can be performed
teratoma) by gently, with a dry cloth, swabbing the surface of the egg.
Phallus inflammation and prolapsed Disinfection by egg dipping or spraying can be performed
Data from references 3, 5, 6, 9–12. using various commercial disinfectants (e.g., dilute chlorine,
iodine, or quaternary ammonium compound solutions).
Possible detrimental effects of egg dipping include damage
or removal of the cuticle layer. The cuticle somewhat seals
the pores and is useful in reducing moisture losses and in
from incubation and hatching to hand-rearing is essential as preventing bacterial penetration of the egg shell. The use of
part of biosecurity protocols. At all stages of development, it formaldehyde for egg dipping is not recommended because
is critical to have the ability to control temperature, humid- of the potential for toxicity to the developing embryo.18
ity, and airflow. Strict sanitation protocols must be used After the eggs have been collected and disinfected, they can
to minimize the movement of infectious agents that may be stored until ready for incubation. Incorrect storage of
lead to mortality of eggs and chicks. Biosecurity protocols eggs can have significant impact on hatching. The storage
should include restriction of personnel, foot baths, clean room should have a temperature range of 12.8°C–15.6°C
clothing, gloves, and a rigorous hygiene plan of the rooms to prevent the onset of embryonic development (which does
and equipment (chemical compounds, ultraviolet radiation, not begin until the egg temperature is greater than 21°C).
ozonization). The facility will be fully operational after all The relative humidity should be maintained at 70%–80%.
the necessary equipment, standard operating procedures, The eggs should not be stored more than 7 days, and it
and personnel training have been established.13–15 should be done in such a way that the air chamber is facing
Two key factors in the success of artificial incubation are upward (there is no need to rotate the eggs in storage).13
fertility and hatchability. Fertility is defined as the ratio of Environmental factors are critical to the normal develop-
fertile eggs to total eggs laid, and hatchability is the ratio of ment of the embryo during the incubation and hatching
eggs that hatch to the total number of fertile eggs. Factors processes. These factors include incubation temperature and
affecting both should be considered when evaluating eggs humidity, egg orientation, egg turning, and ventilation.
that do not hatch and may be divided into three periods: The appropriate incubation temperature for different
prior to oviposition, preincubation, and incubation.15 In avian species is reported and outlined in the literature.15,19
the first period, causes of infertility include: behavioral During incubation, a continuous variation of 0.5°C in the
(e.g., immaturity, pair incompatibility, sexual inexperience, dry bulb temperature will result in a significant increase
improper imprinting, infrequent mating); environmental in congenital malformation and embryo mortality.20 Pro-
(e.g., incorrect photoperiod, incorrect design of the nest box longed temperature extremes beyond that recommended for
or nesting materials, improper design of the enclosure, lack the species can have significant effects on embryo develop-
of visual barriers); and medical (e.g., obesity, endogamy, ment. At higher temperatures, embryonic development will
musculoskeletal, neuromuscular or reproductive disease, be accelerated at different rates and in different tissues. If
nutritional deficiencies or excesses, parasitic disease).3 chicks hatch, they are likely to be reduced in size. At low
CHAPTER 68 Selected Medical Aspects of Bird Reproduction in Ex Situ Conservation 483
temperatures, embryonic development will be retarded, also chicken embryos, and use this information as a basis to
at different rates in different tissues. Slight temperature understand the development in other bird species (Table
deviations are not usually lethal, unless it is due to an exces- 68.1).24–26 A number of techniques may be used for assess-
sive degree or duration. The chicks often have incomplete ing embryonic health via external egg appearance. These
yolk sac retraction or partially open umbilical seals. In these techniques include candling, egg monitoring, egg weight
cases, although embryos may survive the early stages, some loss management, and floating the egg.
late mortality is likely to occur.13 Candling is the primary method for monitoring embry-
Avian eggs lose 15% ± 3% of their weight during incuba- onic development. This method is based on the visualization
tion due to water evaporation through the pores of the of the internal structures of the egg when illuminated with
eggshell. Because this is a physical process (not a metabolic a powerful light (preferably cool) inside a dark room. In
process), it is not affected by the stage of embryonic devel- general, to assess fertility of the egg, candling may be per-
opment and follows a linear pattern throughout incubation. formed at approximately 7–10 days post incubation. Before
Under artificial incubation conditions, this weight loss is this stage, the movement and temperature fluctuations
managed by controlling the humidity in the incubator.13 associated with candling may cause embryonic mortality.27
Domestic poultry are normally incubated at 55%–60% After this stage, candling can be done every 48 hours to
relative humidity, and this is a good starting point for most assess continued embryo growth. In a healthy embryo,
nondomestic species. However, there are species with more candling is usually sufficient once a week.27 This technique
specific needs for humidity requirements; for example, very is useful for detecting very early blood vessels, small cracks
low values (25%–28%) are necessary for ostriches (Struthio and other defects in the shell that should be recorded and,
camelus)21 and higher (84%–86%) values are needed for eventually, repaired. In addition, it is important to detect
roseate spoonbills (Platalea ajaja).22 To evaluate weight loss, early embryo death as evidenced by the presence of a blood
the egg is weighed at the beginning and then at least once or ring. Any eggs with a dead embryo must be removed from
twice a week during the incubation process. If the humidity the incubator so as not to be a continuous culture medium
is too high, weight loss will be insufficient and embryos are for bacteria or fungi. Other uses for this technique are to
likely to become edematous, malpositioned, and/or have define and mark the air cell to assess drawdown and to
residual albumen or fluids that could result in drowning. In define the approach to enter the egg in an assisted hatch or
addition, some may present with unretracted yolk sacs, and/ for its necropsy.27 This technique is not appropriate in thick-
or open umbilical seals are also possible. When humidity shelled or pigmented eggs (e.g., cranes). In these species,
is too low, eggs will undergo excessive weight loss that floating the egg is an acceptable alternative. This method
could cause poor bone mineralization, through alteration of can be used to determine fertility and viability of eggs after
calcium transport, and weak, dehydrated chicks.13 approximately 21 days of incubation. At this stage, eggs float
The absence of egg turning has been shown to result in nearly vertically, with the large end up. From 21 to 23 days,
the adhesion of the embryo to the inner shell membrane. only a slight rotational movement of the egg is noticeable.
Premature or abnormal adhesion of the embryonic mem- Close to hatching, twitching and stronger movements are
branes to the inner shell membrane or other structures can apparent. To pursue this technique, it is necessary to float
increase the incidence of malpositioning, decrease albumen eggs in a mild disinfectant solution (10% povidone-iodine)
use, cause abnormal fluid distribution, decrease oxygen at 43°C and observe the egg for movement for 1 minute
exchange at the surface of the chorioallantois, or lead to a or less. It is not recommended to float the egg for long
poorly developed yolk sac.23 The eggs positioned with the periods of time because this increases the risk of asphyxia-
air cell up hatch best when they are turned to 90 degrees, tion and overheating of the embryo. In addition, it is not
resting at a 45-degree angle, whereas those set horizontally recommended to float the eggs in cool water because the
hatch best when turned approximately to 180 degrees, egg contents will contract and may draw bacteria through
alternating sides. Horizontally set eggs turned in only one the shell.28
direction will cause rupture of the membranes and blood Other monitoring techniques include a digital egg monitor
vessels, resulting in embryo mortality. Setting eggs with the called “Buddy” (Avian Biotech International, Tallahassee,
air cell down causes embryonic malposition.23 Florida). This unit uses infrared beams to detect blood flow
Proper ventilation and gas exchange is another key com- within the developing egg (indicates the detection of heart
ponent of artificial incubation. Maintaining an adequate beat and pulse rate of the chick). The monitor appears to
egg concentration inside the incubator will help to maintain be excellent for assessing viability of the near-hatch chick
normal levels of oxygen and carbon dioxide inside the but is not functional for all avian eggs. Xeroradiography of
incubator as the embryos breathe throughout incubation ostrich eggs is another method that has been reported.29
period.13 The chick embryo assumes the proper position to pip
An understanding of embryo development is important (penetrate) the air cell 2–3 days before hatch.30 Recognizing
for assessing its health. Fifty-five stages of the chicken the normal positioning of the embryo before hatching will
embryo development have been classified and described in aid in the decision making for the need for hatching assis-
detail (Hamburger and Hamilton Stage—H&H stage).24 tance. Seven malpositions with different degrees of embryo
It is possible to extrapolate these stages of development of lethality have been described (Table 68.2).
484 SE C T I O N 12 Avian
Each unhatched egg must be examined postmortem to examination of membranes in situ, sampling for bacterial
document potential causes for lack of hatching. This infor- and fungal culture, followed by exposure of the internal
mation can then be used in the decision-making process of egg contents (Fig. 68.1). Embryonic death is classified
evaluating the artificial incubation protocols. An egg nec- into three stages: early dead embryo (EDE—H&H stage
ropsy technique has been described.31 The first step is the 1–19), mid-dead embryo (MDE—H&H stage 20–39),
TABLE
68.1 Avian Embryonic Development and Possible Causes of Embryo Death
TABLE
68.1 Avian Embryonic Development and Possible Causes of Embryo Death—cont’d
TABLE
68.2 Types of Late Stage Embryo Malpositions
Description of correct positions: Head at large end near air cell, head tucked under right wing upside down
Avian embryo malpositions.
and late dead embryo (LDE—H&H stage 40–45). Dif- the viral causes, herpes viruses and avian paramyxovirus
ferent types of causes that are related to possible death are the most commonly reported. Toxic causes of egg
are associated with the different development stages (see death include oil, insecticides, herbicides, nicotine, and
Table 68.1). Bacteria associated with embryonic death selenium.27 Antibiotic use, including chloramphenicol,
include species of Salmonella, Mycoplasma, Staphylococ- penicillin, tetracycline, oxytetracycline, aminoglycosides,
cus, Chlamydophila, and E. coli. Of the fungal causes, and sulfas, has also been implicated in embryonic
Aspergillus is the most common cause of egg death. Of death.27
486 SE C T I O N 12 Avian
A B C
D E F
• Figure 68.1 Hyacinth macaw (Anodorhynchus hyacinthinus) egg necropsy with malposition 5. (A)
Pip-to-hatch from the middle of the egg and not the air cell. (B) Creating an aperture with scissors for
the air cell. (C) Malposition 5 (upside down). (D) Extraction of the dead embryo. (E) Prominent head and
neck edema. (F) Failure to absorb the yolk sac. (Photography courtesy Temaiken Foundation.)
• BOX 68.2 Common Problems of Neonates exaggerated adrenocortical stress responsiveness, Poult Sci 88:
1352–1357, 2009.
Failure to absorb the yolk sac 5. Bowles H: Evaluating and treating the reproductive system. In
Stunted growth Harrison GJ, Lightfoot T, editors: Clinical avian medicine (vol
Leg and toe deformities II). Palm Beach, FL, 2006, Spix Publishing, pp 519–539.
Constricted toes 6. Crosta L, Gerlach H, Bürkle M, et al: Physiology, diagnosis, and
Stifle subluxation diseases of the avian reproductive tract, Vet Clin North Am Exot
Beak malformations (lateral deviation or mandibular prognathism)
Anim Pract 6(1):57–83, 2003.
Break trauma
Regurgitation
7. Krautwald-Junghanns ME: Ultrasonographic diagnosis of avian
Esophageal or pharyngeal trauma urogenital disorders. In Proceedings of the American Association
Crops stasis of Avian Veterinarians 2005, pp 40–44.
Crop burns and fistulas 8. Wilson HR: Effects of maternal nutrition on hatchability, Poult
Air in the crop Sci 76(1):134–143, 1997.
Foreign body ingestion or impaction 9. Bowles HL: Reproductive diseases of pet bird species, Vet Clin
Aspiration/brooder pneumonia North Am Exot Anim Pract 5(3):489–506, 2002.
Eyelid malformation 10. Carrasco DC, González MS: Reproductive disorders in commonly
Occluded ear opening kept fowl, Vet Clin North Am Exot Anim Pract 20(2):509–538,
Malformed feather or feather stress bars
2017.
Intestinal intussusceptions
Hepatic hepatomas
11. Hadley TL: Management of common psittacine reproductive
Gout disorders in clinical practice, Vet Clin North Am Exot Anim Pract
Wine-colored urine 13(3):429–438, 2010.
Hepatic lipidosis 12. Scagnelli AM, Tully TN: Reproductive disorders in parrots, Vet
Neck deformities Clin North Am Exot Anim Pract 20(2):485–507, 2017.
Congenital abnormalities 13. Kasielke S: Incubation of eggs. In Gage LJ, Duerr RS, editors:
Viral diseases (polyoma; psittacine beak and feather; Hand-rearing birds, Ames, IA, 2007, Blackwell, pp 39–54.
proventricular dilatation disease; Pacheco disease [herpes]; 14. Olsen GH, Clubb SL: Embryology, incubation and hatching. In
pox virus) Harrison GJ, Harrison LR, Ritchie BW, editors: Avian medicine
Microbial diseases (gram-negative infections with E. coli,
and surgery, Philadelphia, PA, 1997, Saunders, pp 54–72.
Klebsiella sp., Enterobacter sp., Salmonella sp. and
Pseudomonas sp.). Chlamydophila sp.
15. Sutherland-Smith M, Witman P: Prehatch protocols to improve
Parasitic diseases (Trichomona, Giardia, Atoxoplasma) hatchability. In Miller RE, Fowler ME, editors: Fowler’s zoo and
Fungal diseases (Candida sp.) wild animal medicine current therapy (vol 7). St. Louis, MO,
2011, Elsevier Health Sciences, pp 324–328.
Data from references 2, 32, 33.
16. Thermote L: Effective hygiene within the hatchery, International
Hatchery Practice 20:18–21, 2006.
17. Lábaque MC, Navarro JL, Martella MB: Microbial contamina-
to address the clinical signs following the basic principles of tion of artificially incubated Greater Rhea (Rhea americana) eggs,
avian emergency medicine and ensuring that the underlying Br Poult Sci 44(3):355–358, 2003.
cause is identified and corrected.34,35 Box 68.2 details the 18. Cadirci S: Disinfection of hatching eggs by formaldehyde
pathologies most frequently observed in neonates. If the fumigation—a review, Arch Geflügelk 73(2):116–123, 2009.
19. Kuehler CM, Good J: Artificial incubation of bird eggs at the
purpose of the hand-rearing is to reintroduce the bird to
Zoological Society of San Diego, Int Zoo Yearb 29:118–136,
its natural habitat, it is recommended to limit as much 1990.
human contact as possible during the different rearing 20. Rideout BA: Investigating embryo deaths and hatching failure,
steps, especially during feeding, because this will decrease Vet Clin North Am Exot Anim Pract 15(2):155–162, 2012.
the chances of imprinting. The use of special puppets or 21. Cooper RG: Handling, incubation, and hatchability of ostrich
clothing for this purpose has been successful. (Struthio camelus var. domesticus) eggs: a review, J Appl Poul Res
10(3):262–273, 2001.
22. Hudson L, Wise J, Tucker R, et al: Development of hand-rearing
References techniques for roseate spoonbills, Ajaia ajaja, at the Fort Worth
Zoo, in Proceedings of the Second Conference of the Nutrition
1. Collar NJ, Butchart SHM: Conservation breeding and avian Advisory Group of the American Zoo and Aquarium Association,
diversity: chances and challenges, Int Zoo Yearb 48(1):7–28, 1997.
2014. 23. Wilson HR: Incubation and hatching of ratites. University of
2. LaBonde J: Avian reproductive and pediatric disorders. In Florida Cooperative Extension Service, Institute of Food and
Proceedings of the American Association of Avian Veterinarians Agriculture Sciences, EDIS, 1996.
2006, pp 229–237. 24. Hamburger V, Hamilton HL: A series of normal stages in
3. Joyner KL: Theriogenology. In Harrison BR, Harrison L, editors: the development of the chick embryo, J Morphol 88:49–89,
Avian medicine: principles and application, Lake Worth, FL, 1994, 1951.
Wingers, pp 748–804. 25. Kasielke S: Proceedings of the Avian Egg Incubation Workshop
4. Schmidt JB, Satterlee DG, Treese SM: Maternal corticosterone 2003. Africam Safari, Puebla, Mexico.
reduces egg fertility and hatchability and increases the numbers 26. Romanoff AL: Critical periods and causes of death in avian
of early dead embryos in eggs laid by quail hens selected for embryonic development, Auk 66(3):264–270, 1949.
488 SE C T I O N 12 Avian
27. Chitty J, Heatley JJ: Diagnosing the unhatched egg. In Proceed- 31. Joyner KL, Abbott UK: Egg necropsy techniques. In Proceed-
ings of the American Association of Avian Veterinarians 2011, ings of the American Association of Avian Veterinarians, 1991:
pp 109–117. 146–152.
28. Gabel RR, Mahan TA: Incubation and hatching. In Ellis DH, 32. Flammer K, Clubb SL: Neonatology. In Harrison BR, Harrison
Gee GF, Mirande CM, editors: Cranes: their biology, husbandry L, editors: Avian medicine: principles and application, Lake Worth,
and conservation, Baraboo, WI, 1996, US Department of the FL, 1994, Wingers, pp 805–841.
Interior, National Biological Service, Washington, DC, and 33. Romagnano A: Psittacine incubation and pediatrics, Vet Clin
International Crane Foundation, pp 59–76. North Am Exot Anim Pract 15(2):163–182, 2002.
29. Ley DH, Morris RE, Smallwood JE, Loomis MR, et al: Mortal- 34. de Matos R, Morrisey JK: Emergency and critical care of
ity of chicks and decreased fertility and hatchability of eggs small psittacines and passerines, Semin Avian Exot Pet Med
from a captive breeding pair of ostriches, J Am Vet Med Assoc 14(2):90–105, 2005.
189(9):1124, 1986. 35. Riley J, Barron H: Wildlife emergency and critical care, Vet Clin
30. Freeman B, Vince M: Physiology of hatching. In Development of North Am Exot Anim Pract 19(2):613–626, 2016.
avian embryo: a behavioral and physiological study, London, 1974,
Chapman and Hall, pp 249–260.
SECTION 13
Marsupials
69 Tasmanian Devil Facial Tumor Disease, 490
489
69
Tasmanian Devil Facial Tumor Disease
CAROLYN J. HOGG AND KATHERINE BELOV
T
he Tasmanian devil (Sarcophilius harrisii) is the of starvation as the tumor destroys facial bones and dental
world’s largest marsupial carnivore. Devil popula- arcades. An immune response to DFTD has been noted
tions have declined by up to 95% in some areas of in six wild devils at West Pencil Pine (northern Tasmania),
the Australian island state of Tasmania since the emergence with four of these devils showing signs of tumor regression,
of the infectious cancer, devil facial tumor disease (DFTD) although these incidences are extremely rare.7 At this time
in 1996.1 Since that time, a second clonal, infectious cancer there is no treatment for DFTD, although new immuno-
in Tasmanian devils has been described, known as DFT2,2 therapy trials are giving promising results.8
with the original form known as DFT1.
Cause
Signs and Symptoms
DFT1 is a peripheral nerve sheath tumor that arose from
DFT1 tumors present as large, solid, soft tissue masses that a Schwann cell or Schwann cell precursor, because tumors
ulcerate, first appearing on the head and/or neck regions produce the Schwann cell–specific myelio protein, periaxin.9
(Fig. 69.1). Histologically, they form subepithelial expansile Due to its recent discovery, the epidemiology of DFT2 is cur-
masses of round spindloid cells with abundant eosinophilic rently unknown. Tasmanian devils have 14 chromosomes.
cytoplasm encased within a pseudocapsule. The tumor is The cytogenetic profile of DFT1 differs markedly from the
locally aggressive and metastasizes in 65% of cases, 57% normal devil karyotype and is characterized by the absence
of these to the local lymph nodes. Tumors similar to those of identifiable chromosome 2, missing X and Y sex chromo-
on the face may occur later in other parts of the body.1 somes, and the presence of four marker chromosomes. The
In contrast, the tumors from DFT2 are characterized by DFT1 tumor is evolving over time, with a number of dif-
sheets of pleomorphic (amorphic to stellate and fusiform) ferent strains now present.10 DFT1 tumor studies have the
cells arranged in a solid pattern. Periaxin is a known histo- same chromosomal rearrangements, indicating their origin
chemical marker that is diagnostic for DFT13; DFT2 tumors as clones from a rogue cell line and transferred between
appear to be negative for periaxin, an immunohistochemical individuals as allografts.11 The DFT2 cytogenetic profile is
marker that is diagnostic for DFT1.2 distinct from DFT1, in which all tumors described showed
Male and female devils are equally affected by DFTD, identical structural abnormalities, including the presence of
with animals younger than 2 years rarely affected. It is additional material on chromosomes 1, 2, and 4, a deletion
possible young animals are bitten less often or do not involving chromosome 5, and monosomy for chromosome
develop tumors due to the length of the incubation period 6. Both X and Y sex chromosomes are present.2
of the disease. However, it has also been proposed that In theory, because the tumor cells are different from the
young devils are protected from DFTD by the presence of host cells, they should be recognized as foreign and rejected.
antimicrobial peptides in mother’s milk and pouch,4 as well However, this is not the case, with lymphocyte infiltration
as the interplay between puberty and the devil’s immune into DFTD tumors rarely observed. Research has shown
system.5 In areas where the disease has been present for a that devils do have a competent immune system, similar
long period of time, populations of Tasmanian devils are to that of other mammals.12 Initially, it was thought that,
still persisting (Save the Tasmanian Devil Program [STDP], due to low genetic diversity at the major histocompatibility
personal communication). complex (MHC) locus, the devil’s immune system would
Once clinical, the course of DFTD is rapid, with tumors not recognize the tumor as being foreign.13 However, recent
enlarging from small nodules to large friable masses over the research has shown that the tumor is able to downregulate
course of 2–3 months, and death usually occurs within 6 its MHC so it is undetectable by the devil’s immune
months.6 Mortality is almost always 100%, mostly because system.14
490
CHAPTER 69 Tasmanian Devil Facial Tumor Disease 491
Acknowledgments
Thanks to Peter Holtz, the STDP staff, and other research-
ers who work tirelessly on the program to save the devil,
and all the staff and management of the zoos and wildlife
facilities, both national and international, who contribute
significantly to the Tasmanian Devil Insurance Population.
CJH & KB are funded by the Australian Research Council
• Figure 69.2 Tasmanian devil (Sarcophilius harrisii) released onto
Maria Island as part of the metapopulation in 2012. and the STDP Appeal.
15. Hamede RK, McCallum H, Jones M: Biting injuries and trans- 28. Cheng Y, et al: Low MHC class II diversity in the Tasmanian
mission of Tasmanian devil facial tumour disease, J Anim Ecol devil (Sarcophilus harrisii), Immunogenetics 64(7):525–533,
82(1):182–190, 2013. 2012.
16. Guiler E: Observations on the Tasmanian devil, Sarcophilus har- 29. Ujvari B, et al: Telomere dynamics and homeostasis in a trans-
risii (Marsupialia : Dasyuridae) II. Reproduction, breeding and missible cancer, PLoS ONE 7(8):e44085, 2012.
growth of pouch young, Aust J Zool 18(1):63–70, 1970. 30. Cheng Y, Belov K: Characterisation of non-classical MHC class I
17. Pyecroft S, et al: Towards a case definition for devil facial tumour genes in the Tasmanian devil (Sarcophilus harrisii), Immunogenet-
disease: what is it?, Ecohealth 4(3):346–351, 2007. ics 66(12):727–735, 2014.
18. Hogg CJ, et al: Metapopulation management of an endangered 31. Ujvari B, et al: Anthropogenic selection enhances cancer evo-
species with limited genetic diversity in the presence of disease: lution in Tasmanian devil tumours, Evol Appl 7(2):260–265,
the Tasmanian devil Sarcophilus harrisii, Int Zoo Yearb 51:1–17, 2014.
2016. 32. Cui J, Cheng Y, Belov K: Diversity in the toll-like receptor genes
19. Cheng Y, et al: Significant decline in anticancer immune capacity of the Tasmanian devil (Sarcophilus harrisii), Immunogenetics
during puberty in the Tasmanian devil, Sci Rep 7:44716, 2017. 67(3):195–201, 2015.
20. Lachish S, McCallum H, Jones M: Demography, disease and the 33. Ujvari B, et al: Immunoglobulin dynamics and cancer prevalence
devil: life-history changes in a disease-affected population of Tas- in Tasmanian devils (Sarcophilus harrisii), Sci Rep 6:25093,
manian devils (Sarcophilus harrisii), J Anim Ecol 78(2):427–436, 2016.
2009. 34. Jones ME, et al: Conservation management of tasmanian devils
21. McCallum H, et al: Transmission dynamics of Tasmanian devil in the context of an emerging, extinction-threatening disease:
facial tumor disease may lead to disease-induced extinction, Devil facial tumor disease, Ecohealth 4(3):326–337, 2007.
Ecology 90(12):3379–3392, 2009. 35. CBSG: Tasmanian Devil PHVA Final Report, 2008. IUCN/SSC
22. Fancourt BA, et al: Devil declines and catastrophic cascades: Conservation Breeding Specialist Group: Apple Valley, MN.
is mesopredator release of feral cats inhibiting recovery of the 36. Farquharson KA, Hogg CJ, Grueber CE: Pedigree analysis
Eastern Quoll?, PLoS ONE 10(3):e0119303, 2015. reveals a generational decline in reproductive success of captive
23. Doherty TS, et al: Impacts and management of feral cats (Felis Tasmanian devil (Sarcophilus harrisii): implications for captive
catus) in Australia, Mammal Review 47(2):83–97, 2017. management of threatened species, J Hered 108(5):488–495,
24. Miller W, et al: Genetic diversity and population structure of the 2017.
endangered marsupial Sarcophilus harrisii (Tasmanian devil) (vol 37. Hogg CJ, et al: Influence of genetic provenance and birth origin
108, pp 12348, 2011), Proc Natl Acad Sci USA 109(45):18625– on productivity of the Tasmanian devil insurance population,
18625, 2012. Conservation Genetics 16(6):1465–1473, 2015.
25. Jones ME, et al: Genetic diversity and population structure 38. Gooley R, et al: No evidence of inbreeding depression in a Tas-
of Tasmanian devils, the largest marsupial carnivore, Mol Ecol manian devil insurance population despite significant variation
13(8):2197–2209, 2004. in inbreeding, Sci Rep 7:1830, 2017.
26. Storfer A, et al: Landscape genetics of the Tasmanian devil: 39. Grueber CE, et al: Increasing generations in captivity is associ-
implications for spread of an infectious cancer, Conserv Genet ated with increased vulnerability of Tasmanian devils to vehicle
18(6):1287–1297, 2017. strike following release to the wild, Sci Rep 7:2161, 2017.
27. Kreiss A, et al: Allorecognition in the Tasmanian devil (Sarcophi- 40. Hogg CJ, et al: “Devil tools & tech”: a synergy of conservation
lus harrisii), an endangered marsupial species with limited genetic research and management practice, Conserv Lett 10(1):133–138,
diversity, PLoS ONE 6(7):e22402, 2011. 2017.
70
Medical Aspects of Potoroid Marsupial
Conservation Translocations
TIMOTHY J. PORTAS
494
CHAPTER 70 Medical Aspects of Potoroid Marsupial Conservation Translocations 495
TABLE
70.1 Extant Potoroid Species, Their Conservation Status, and Distribution
Restraint, Anesthesia, and Analgesia • Figure 70.1 Short periods of restraint for minimally invasive sam-
pling such as blood collection from the lateral coccygeal vein are
Free-ranging potoroids are typically caught in small, wire, possible with potoroids restrained in soft cloth bags.
cage traps baited with a mixture of rolled oats and peanut
butter. Internal ceiling padding and external visual barriers
are recommended on cage traps to reduce trap-associated Ejection of pouch young is a risk when trapping,
trauma and minimize stress. On removal from traps, poto- handling, and transporting female potoroids. Females with
roids should be placed into soft cloth bags for subsequent large pouch young should be excluded from conservation
handling. Although potoroids will tolerate short periods of translocations, and the pouch of females with furless pouch
manual restraint (up to 10 minutes) for examination and young should be secured with adhesive tape for handling
minimally invasive sampling (Fig. 70.1) without apparent and transportation. The tape is left in situ at release with
adverse effects, anesthesia is recommended to minimize females invariably removing the tape after they have settled.
stress and allow for complete physical examination and In addition, females with two active mammary glands
sample collection. Inhalation anesthesia using isoflurane or should be precluded due to the likelihood of an untrapped
sevoflurane in oxygen administered by mask is the preferred emergent or young at-foot joey that remains dependent
method of anesthetic induction; premedication is gener- on milk.
ally not required, but intramuscular (IM) administration Administration of analgesic drugs is indicated where
of diazepam (1 mg/kg) or midazolam (0.3 mg/kg) may potoroids have sustained trapping-related injuries or been
be considered in fractious or unduly stressed individuals. subjected to invasive sampling procedures during the
Potoroids are recovered from anesthesia in soft cloth bags conservation translocation process. Meloxicam (0.2 mg/kg
but must be monitored and managed to prevent airway subcutaneously (SC) q 24 hours), buprenorphine (0.01 mg/
occlusion until sufficiently recovered. kg IM q 12 hours), and tramadol (5 mg/kg IM q 12 hours),
496 SE C T I O N 13 Marsupials
although unsupported by pharmacokinetic data, appear to possible, collars should be applied and assessed in cap-
be safe and efficacious choices for providing analgesia in tivity, where close monitoring can occur prior to field
potoroids. A topical proprietary preparation containing application.
lignocaine, bupivacaine, epinephrine, and cetrimide has
recently been used to provide hemostasis, analgesia, and Disease Risk Assessment
antisepsis following ear-tagging and collection of ear punch
biopsy samples for genetic analysis in reintroduced eastern Although the principal reason most potoroid conservation
bettongs. translocations fail is predation by introduced predators,
disease has the potential to significantly impact the success
Transportation of these programs.1 A comprehensive disease risk assessment
is therefore recommended but beyond the scope of this
Potoroids are best transported in soft cloth bags suspended chapter. Readers are referred to Chapter 2: Risk Analysis
within adequately ventilated transport containers. Trans- Framework Guidance for Wildlife Health Professionals
portation should be timed to avoid high environmental by Travis and Smith in this volume and several recent
temperatures. Although potoroids have been transported reviews for detailed approaches to conducting disease risk
without the use of sedative or neuroleptic drugs, a study assessments in conservation translocations.11–13 Table 70.2
demonstrated marked elevations in creatine kinase in eastern outlines some known and potential pathogens and parasites
bettongs transported by road and air for reintroduction.9 of potoroids that may warrant consideration during disease
The use of diazepam (1 mg/kg IM) in these animals appears risk assessments for conservation translocations.
to have ameliorated the effects of exertional myopathy.
TABLE Known and Potential Pathogens and Parasites of Potoroids That May Warrant Consideration in Disease
70.2 Risk Assessment During Conservation Translocations
6. Hing S, Northover AS, Narayan EJ, et al: Evaluating stress 13. Jakob-Hoff RM, MacDiarmid SC, Lees C, et al: Manual of
physiology and parasite infection parameters in the translocation procedures for wildlife disease risk analysis. Paris, France, World
of critically endangered woylies (Bettongia penicillata), Ecohealth Organisation for Animal Health, 2014.
14:S128–S138, 2017. 14. Vaughan RJ, Warren KS, Mills JS, et al: Hematological and serum
7. Hing S, Narayan EJ, Thompson RCA, et al: Identifying factors biochemical reference values and cohort analysis in the Gilbert’s
that influence stress physiology of the woylie, a critically endan- potoroo (Potorous gilbertii), J Zoo Wildl Med 40:276–288,
gered marsupial, J Zool 2016, doi:10.1111/jzo.12428. 2009.
8. Northover AS, Godfrey SS, Lymbery AJ, et al: Evaluating the 15. Pacioni C, Robertson ID, Maxwell M, et al: Hematologic char-
effects of ivermectin treatment on communities of gastrointestinal acteristics of the woylie (Bettongia penicillata ogilbyi), J Wildl Dis
parasites in translocated woylies (Bettongia penicillata), Ecohealth 49:816–830, 2013.
14:S117–S127, 2017. 16. Pacioni C, Johansen CA, Mahony TJ, et al: A virological
9. Portas T, Fletcher D, Spratt D, et al: Health evaluation of free- investigation into declining woylie populations, Aust J Zool
ranging eastern bettongs (Bettongia gaimardi) during transloca- 61:446–453, 2014.
tion for reintroduction, J Wildl Dis 50:210–223, 2014. 17. Dickson J, Hopkinson WI, Coackley W, et al: Herpesvirus
10. Portas TJ, Cunningham RB, Spratt D, et al: Beyond morbidity hepatitis in rat kangaroos, Aust Vet J 56:463–464, 1980.
and mortality in reintroduction programmes: changing health 18. Vaughan RJ, Vitali SD, Eden PA, et al: Cryptococcosis in Gil-
parameters in reintroduced eastern bettongs (Bettongia gaimardi), bert’s and long-nosed potoroo, J Zoo Wildl Med 38:567–573,
Oryx 50:674–683, 2016. 2007.
11. Ewen JG, Sainsbury AW, Jackson B, et al: Disease risk manage- 19. Ladds P: Pathology of Australian native wildlife, Melbourne,
ment in reintroduction. In Armstrong DP, Hayward MW, Moro Australia, 2009, CSIRO Publishing.
D, et al, editors: Advances in reintroduction biology of Australian 20. Bennet MD, Reiss A, Stevens H, et al: The first complete
and New Zealand fauna, Melbourne, Australia, 2015, CSIRO Papillomavirus genome characterized from a marsupial host: a
Publishing, pp 43–57. novel isolate from Bettongia penicillata, J Virol 84:5448–5453,
12. Hartley M, Sainsbury A: Methods of disease risk analysis in 2010.
wildlife translocations for conservation purposes, Ecohealth
14:S16–S29, 2017.
71
Macropod Pediatric Medicine
MICHELLE CAMPBELL-WARD
500
CHAPTER 71 Macropod Pediatric Medicine 501
bacteria.14 Pouch young are able to synthesize these soon For animals undergoing hand-rearing, carer or owner
after birth to complement those present in milk.15,16 notes on growth, behavior, feeding, and toileting should be
The development of endothermy in marsupial young inspected. If such notes are not routinely kept, the carer/
begins at least halfway through pouch life concurrent with owner should be encouraged to do so.4 In circumstances
the initiation of thyroid function. In the tammar wallaby where aspiration of formula is suspected or feeding has been
this transition occurs between 55 and 200 days of age at a problematic, inspection of the feeding equipment including
mass of 70–300 g.3 A myriad of morphologic, physiologic, teat and hole size and observing feeding technique can be
and behavioral changes that allow for endothermy and the informative.18 Skin moisturization protocols and artificial
maintenance of body temperature occurs across this time pouch conditions should be assessed for furless joeys. The
period.17 length of time in care may be relevant as orphans lose most
maternal immunoglobulins by 4–6 weeks after separation
Approach to the Pediatric from the mother and are left protected only by an under-
developed active immune system.19
Macropod Consultation
History Physical Examination and Assessment
The clinical evaluation relies heavily on thorough history- The examination should take place in a warm room with
taking. Box 71.1 provides a checklist of important factors pre-warmed hands in the majority of circumstances.
that should be discussed during initial patient assessment. For orphaned pouch-dependent young, the bulk of the
Mimicking the environmental factors necessary for normal examination is best conducted while they are held within an
development may be challenging, especially if the owner or artificial pouch. If the joey is still with the mother, maternal
carer lacks experience. Identifying husbandry deficiencies sedation may be required.
can be instrumental in achieving a diagnosis and developing Estimating age and determining whether the animal’s
a therapeutic plan. behavior is appropriate for the stage of development are
vital steps in the assessment process.20 Knowledge of
• BOX 71.1 History-Taking Checklist for the expected growth trajectories and developmental milestones
Macropod Pediatric Consultation of a given species is necessary for interpretation of findings.
Species-specific growth charts are available online.21
• Signalment The age factor is defined as the age as a proportion of
• Species total expected pouch life, for example, an age factor 0.7
• Sex
• Stage of development, for example, pouch-dependent pouch young has completed 70% of expected pouch life. As
age factor <0.4, 0.4–0.6, 0.7–1.0 (see text); emerged a general rule, the intensity of care necessary for marsupial
(i.e., at-foot); weaned pouch young at an age factor of less than 0.4 or while still
• Weigh and take body measurements in the fixed lactation phase is substantial and the chance of
• Presenting complaint successfully hand-rearing a pouch young from this stage to
• Duration and clinical course (improving, deteriorating,
stable) adulthood is extremely low. If an animal of this size cannot
• Previous medical problems and/or treatments (drugs, dose be returned to the mother’s pouch or cross-fostered (see
and duration) later), euthanasia should be considered.
• If orphaned, reason for orphaning (e.g., maternal death or A thorough physical examination includes measurement
rejection, illness or injury, unknown, intentional) and age/age of vital parameters and assessment of skin health (especially if
factor/weight at time of separation from the mother
• If wild, time since rescue and rescue location furless), hydration status, and gait (if emerging or emerged).
• Housing (e.g., artificial pouch, indoor room, grassed yard)— Critical issues that require immediate stabilization are sum-
obtain detailed description marized in Table 71.1.
• Temperature (of artificial pouch or ambient if emerged) For critically ill animals, obtaining a blood sample for
• Diet glucose and electrolyte assessment is valuable. Generally
• Formula type, dilution rate, volume per feed, frequency of
feeds additional diagnostic tests such as radiography or other
• Method of feeding (bottle-fed, lapping from bowl) imaging, complete blood count, serum biochemistry panel,
• Types and quantities/proportions of solids offered and sampling for cytology and/or culture may be postponed
• Supplements until the stabilization process has commenced.
• Hygiene routine (e.g., frequency of bottle cleaning, pouch In situations where a pediatric patient can be assessed,
changes, disinfectants used)
• Skin care (if hairless) treated, and quickly returned to a healthy mother capable
• Toileting routine of providing adequate care, that is usually preferable to
• Contact with other animals (conspecifics; wild and hand-rearing. Partial temporary closure of the pouch orifice
domestic animals of other species) with tape may reduce the risk of the pouch young being
• Proposed fate of the animal (zoo collection, return to wild, evicted in the immediate recovery phase. This approach
pet)
secures the joey while allowing the dam to be able to access
the pouch for cleaning.
502 SE C T I O N 13 Marsupials
TABLE
71.1 Critical Issues in Macropod Pediatrics That Require Immediate Stabilization
CRI, Continuous rate infusion; IV, intravenous; NG, nasogastric; SC, subcutaneous.
TABLE
71.2 Clinical Signs, Diagnosis, and Treatment of Common Macropod Pediatric Conditions
Continued
504 SE C T I O N 13 Marsupials
TABLE
71.2 Clinical Signs, Diagnosis, and Treatment of Common Macropod Pediatric Conditions—cont’d
*Psychological and environmental stressors include an insecure pouch environment, excessive handling, petting and playing with children, contact with unfamiliar
environments and noises, sudden changes in routine (e.g., prolonged pouch deprivation during the process of pouch weaning), sudden changes in feeding
schedule, sudden withdrawal of contact with carer or buddy. CBC, Complete blood count; GI, gastrointestinal; NG, nasogastric tube.
16. Wang J, Wong ESW, Whitley JC, et al: Ancient antimicrobial 26. Munn AJ, Dawson TJ, Maloney SK: Ventilation patterns in red
peptides kill antibiotic resistant pathogens: Australian mammals kangaroos (Macropus rufus Desmarest): juveniles work harder
provide new options, PLoS ONE 6(8):e24030, 2011. than adults at thermal extremes, but extract more oxygen per
17. Andrewartha SJ, Cummings KJ, Frappell PB: Acid-base balance breath at thermoneutrality, J Exp Biol 210:2723–2729, 2007.
in the developing marsupial: from ectotherm to endotherm, J 27. Parameswaran N, O’Handley RM, Grigg ME, et al: Vertical
Appl Physiol 116(9):1210–1219, 2014. transmission of Toxoplasma gondii in Australian marsupials,
18. McCracken H: Veterinary aspects of hand-rearing orphaned mar- Parasitology 136(9):939–944, 2009.
supials. In Vogelnest L, Woods R, editors: Medicine of Australian 28. Cripps J, Beveridge I, Ploeg R, et al: Experimental manipula-
mammals, Collingwood, Australia, 2008, CSIRO, pp 13–37. tion reveals few subclinical impacts of a parasite community in
19. Old JM, Deane EM: Development of the immune system and juvenile kangaroos, Int J Parasitol Parasites Wildl 3(2):88–94,
immunological protection in marsupial pouch young, Dev Comp 2016.
Immunol 24:445–454, 2000. 29. Taggart DA, Schultz DJ, Fletcher TP, et al: Cross-fostering and
20. Vogelnest L, Portas T: Macropods. In Vogelnest L, Woods R, short-term pouch isolation in macropodoid marsupials: implica-
editors: Medicine of Australian mammals, Collingwood, Australia, tions for conservation and species management. In Eldridge CG,
2008, CSIRO, pp 133–225. editor: Macropods: the biology of kangaroos, wallabies and rat-
21. Wombaroo: Growth and feed charts for macropods. Retrieved kangaroos, Collingwood, Australia, 2010, CSIRO, pp 263–278.
from http://www.wombaroo.com.au/native-wildlife/kangaroo, 30. Hetz JA, Menzies BR, Shaw G, et al: Effects of nutritional
2017. manipulation on body composition in the developing marsupial,
22. McKenzie S, Deane EM, Burnett L: Haematology and serum Macropus eugenii, Mol Cell Endocrinol 428:148–160, 2016.
biochemistry of the tammar wallaby Macropus eugenii, Comp 31. McLelland DJ, Fielder K, Males G, et al: Successful transfer of
Clin Path 11:229–237, 2002. a Goodfellow’s tree kangaroo (Dendrolagus goodfellowi) pouch
23. Vogelnest L: Marsupialia (marsupials). In Miller RE, Fowler ME, young to a yellow-footed rock wallaby (Petrogale xanthopus)
editors: Fowler’s zoo and wild animal medicine (vol 8). St. Louis, surrogate, Zoo Biol 34(5):460–462, 2015.
MO, 2015, Elsevier, pp 255–274. 32. American Veterinary Medical Association: AVMA guidelines for
24. Smits A, Kulo A, de Hoon JN, et al: Pharmacokinetics of drugs the euthanasia of animals. Schaumburg, Illinois, 2013.
in neonates: pattern recognition beyond compound specific 33. National Resource Management Ministerial Council: National
observations, Curr Pharm Des 18(21):3119–3146, 2012. Code of Practice for the Humane Shooting of Kangaroos and
25. Barnes TS, Goldizen AW, Coleman GT: Hematology and serum Wallabies for Commercial Purposes. Canberra, Department of
biochemistry of the brush-tailed rock-wallaby (Petrogale penicil- the Environment, Water, Heritage and the Arts, 2008.
lata), J Wildl Dis 44(2):295–303, 2008.
SECTION 14
Small Mammals
72 White-Nose Syndrome: Cutaneous Invasive
Ascomycosis in Hibernating Bats, 508
507
72
White-Nose Syndrome: Cutaneous
Invasive Ascomycosis in
Hibernating Bats
CAROL U. METEYER AND MICHEL LE L. VERANT
508
CHAPTER 72 White-Nose Syndrome: Cutaneous Invasive Ascomycosis in Hibernating Bats 509
• Figure 72.1 Detection of white-nose syndrome and the causative agent, Pseudogymnoascus destruc-
tans, as of October 12, 2017. The black dot surrounded by a red circle is the site of initial mortality.
(Updates at https://www.whitenosesyndrome.org/resources/map.)
WNS.18–20 Once bats become euthermic following spring indicating that this may not be the case for all species or
emergence from hibernation, inflammation may be severe, locations.
and result in further damage to the wing membrane.21 The invasion of wing membrane and replacement of
Differences in susceptibility to WNS infection and tissue by Pd leads to a complex suite of behavioral and
disease severity have been observed across species of physiologic disturbances that may ultimately lead to
cave-hibernating bats. Some data suggest that big brown death.25 Increased frequency of arousal from torpor has been
bats are resistant to WNS, and population increases for demonstrated in hibernating bats affected by WNS.26,27
this species have been reported within regions affected by These arousals are thought to contribute to higher energy
WNS.22,23 Differences in species susceptibility to WNS may demands, depletion of energy reserves prior to spring
be correlated with temperature of selected hibernation sites, emergence, and emaciation often seen in bats with WNS.
colony size and tendency to cluster during hibernation, Other life-threatening physiologic disruptions reported in
body size and hibernation physiology, native microbial skin bats with WNS include electrolyte imbalances, dehydra-
communities, and the composition of sebaceous lipids on tion, and acid-base disturbances, which are also thought to
the skin surface of the bat. It is likely that a combination of contribute to mortality.28–30
factors is responsible for this variable sensitivity to infection
with Pd and expression of disease in species and populations Clinical Signs
surviving in areas with WNS.
As WNS spreads, mortality of infected bats at lower Observation of bats flying during the day or on the ground
latitudes and warmer climates may be moderated due to outside of hibernacula in colder winter climates is pre-
shorter hibernation seasons and more frequent foraging liminary evidence that WNS is present in the population.
during winter. However, recent studies in Tennessee have Bats may also be seen moving closer to the entrance of
documented population declines of up to 98% in hibernat- hibernacula, flying into buildings, trembling on the ground,
ing colonies of northern long-eared bats (Myotis sodalis),24 or struggling to fly.6,25 Healthy hibernating bats in southern
510 SE C T I O N 14 Small Mammals
• Figure 72.2 Muzzle of a little brown bat (Myotis lucifugus) with white
lit from above with hand-held 51 LED 385-nm ultraviolet flashlight
fungal growth confirmed to be white-nose syndrome by histopathol- shows points of orange–yellow fluorescence consistent with sites of
ogy. Presence of Pseudogymnoascus destructans was confirmed by white-nose syndrome lesions. (Modified from Figure 1E; Turner GG,
culture and molecular methods. (Photograph by Carol Meteyer, US Meteyer CU, Barton H, et al: Nonlethal screening of bat-wing skin
Geological Survey.) with the use of ultraviolet fluorescence to detect lesions indicative of
white-nose syndrome. J Wildl Dis 50:566–573, 2014.)
latitudes normally forage outside of hibernacula during
winter, but colonies of bats with WNS are more active,
depart hibernacula in higher numbers earlier in the day,
and can demonstrate other behaviors similar to bats affected
with WNS in northern locations.31
Delicate white hyphae are often seen on exposed skin
surfaces of hibernating bats with WNS in hibernacula (Fig.
72.2). However, other noninvasive dermatophytes on bats
can produce similar white hyphae,32 so further study is
warranted to confirm WNS. In addition, bats with WNS
may not have visible fungus,33 and handling or removing an
infected bat from a hibernaculum generally results in loss
of visible hyphae on the skin surface.17
Wing membranes are the most common site of Pd infec- • Figure 72.4 Histologic section of wing membrane from the little
tion, but with the exception of readily observable visible brown bat (Myotis lucifugus) in Fig. 72.2. Periodic acid-Schiff stains
hyphae that may or may not be present, other gross signs hyphae of Pseudogymnoascus destructans magenta. Branching,
of WNS are often subtle. For example, extended wing septate hyphae with irregularly parallel walls fill cup-shaped epidermal
membranes may be stiff or sticky with loss of elasticity and erosions that have a discreet interface with host tissue. Bar = 20 µm.
(Photograph by Carol Meteyer, US Geological Survey.)
areas of tissue contraction. Examining the wing surface
under a long-wave ultraviolet (UVA) light (366–385 nm)
can highlight pinpoint or coalescing areas of orange-yellow agar) incubated at a cold temperature (approximately
fluorescence (Fig. 72.3), indicating potential presence of 5°C–10°C) for a minimum of 2 weeks.2,37 Curved conidia
epidermal lesions caused by Pd.34 However, UVA light produced by Pd are morphologically distinct from other
should be used only as a screening tool to identify individual fungi generally found on bats,38 but identification of the
bats for further diagnostic testing to confirm WNS. In the isolate by Pd-specific PCR or PCR amplification of the
weeks following spring emergence, wing damage such as rRNA gene region and sequence analysis is necessary for
depigmentation, holes, and tears can be observed in bats definitive confirmation.
infected with Pd during hibernation. However, healing of Skin lesions associated with WNS can be unevenly
wing tissue occurs within weeks postemergence.35,36 distributed on the wing, and without targeted biopsy sam-
pling (described in “Surveillance”), a whole carcass should
Diagnosis be submitted for extensive evaluation of ear, muzzle, and
wing membrane.26 The histologic characteristics of dermal
Confirmation of WNS in a bat requires identification of invasion by Pd vary as the disease progresses, but the unique
characteristic histologic changes17 and confirmation of pres- pattern that defines WNS consists of dense aggregates
ence of Pd by real-time polymerase chain reaction (PCR) or of large irregular branching and septate hyphae forming
fungal culture analysis. Pseudogymnoascus destructans may be cupping erosions that have a defined interface at the margin
cultured from samples using suitable culture medium (e.g., with host tissue (Fig. 72.4).17 Evidence of a cellular inflam-
Sabouraud dextrose agar or dextrose-peptone-yeast extract matory response is also generally absent in hibernating
CHAPTER 72 White-Nose Syndrome: Cutaneous Invasive Ascomycosis in Hibernating Bats 511
bats, but euthermic bats collected outside hibernacula in several weeks.35 Bats in the wild also recover if they survive
spring can have abundant inflammation, which completely the potentially fatal wing damage that may occur poste-
resolves without fibrosis or scarring.35 mergence.36 Although bats are generally re-infected with Pd
each fall as they enter hibernation,39 band recoveries have
Surveillance shown survival of bats for up to 6 years in spite of WNS.48
Other potential disease management options that are
Conducting surveillance for WNS in hibernating bats is being tested include vaccination, ultraviolet (UVA) light,
recommended in late winter to decrease disruption of bats antifungal compounds, and biological agents. Some of
during hibernation, and because clinical signs, prevalence these have demonstrated effectiveness against Pd in the
and abundance of Pd, and skin lesions have been shown to laboratory49,50,50a; however, field trials to assess applicability,
increase in bats over time during hibernation.33,39 Selecting safety, and efficacy in wild bats, as well as potential ecologic
bats with visible signs of infection can increase the probabil- side effects, are ongoing.
ity of detecting Pd in a population.33 Noninvasive sampling To date, management actions for reducing impacts of
techniques for Pd include epidermal swabs from wings and WNS on bat populations have primarily focused on reducing
muzzle, and less invasive sampling for histopathology can disturbance of bats through protection of hibernacula, and
be done using wing punch biopsies taken from areas of minimizing risks of human-assisted spread of Pd through
visible white fungus or by targeting areas of orange-yellow education and development of decontamination protocols.
fluorescence under UVA light.40 However, these techniques Additional information and guidance for implementing
have reduced sensitivity for detecting Pd or WNS, and these management actions may be found in Recommen-
whole carcasses of fresh dead bats should be submitted to dations for Managing Access to Subterranean Bat Roosts to
an experienced diagnostic laboratory for testing if clinical Reduce the Impacts of White-Nose Syndrome in Bats, and the
signs are seen in new geographic areas or in species for National White-Nose Syndrome Decontamination Protocol.
which WNS has not be previously diagnosed. These guidance documents and additional acceptable
Sampling bats as they emerge from hibernation in spring management practices for bats are available on the White-
is also possible, but the probability of detecting Pd or WNS Nose Syndrome Response Plans web site (https://www
declines over the first few weeks posthibernation as wings .whitenosesyndrome.org/white-nose-syndrome-response
heal and the amount of Pd on bats declines. Observation -plans).
of wing damage after spring emergence may reflect presence Alterations of habitats used by bats for foraging or roost-
of WNS but is not specific for the disease.41,42 Although ing have also been proposed as management actions to
probability of detecting Pd on bats during summer months increase survival and to support recovery of bat populations
is low, Pd has been detected on bats that use caves and mines susceptible to WNS.51 Given the long life spans and low
for day roosts in summer,11,42a and Pd is known to persist reproductive rates of bats, adult and juvenile survival are
in the physical environment of hibernacula year-round with critically important for long-term population recovery.47
no seasonal differences in detection probability.43 Detection However, further research is needed on the implementation
of Pd in the environment of a hibernaculum is correlated and effectiveness of these approaches for reversing popula-
with presence of WNS in bats at those sites, but sensitivity tion declines in bat species affected by WNS.
of environmental sampling for detecting Pd is dependent
on the design of the sampling scheme and can lag in
time behind first detection of Pd in bats at a site.42b,43 For References
updated information on Pd/WNS surveillance, see the US
A full WNS bibliography can be found at, https://www.whitenose-
Geological Survey National Wildlife Health Center White- syndrome.org/wns-bibliography
Nose Syndrome web page (https://www.nwhc.usgs.gov/ 1. Blehert DS, Hicks AC, Behr MJ, et al: Bat white-nose syndrome:
disease_information/white-nose_syndrome/index.jsp). an emerging fungal pathogen? Science 323:227, 2009.
2. Gargas A, Trest MT, Christensen M, et al: Geomyces destructans
Disease Mitigation sp. nov. asssociated with bat white-nose syndrome, Mycotaxon
108:147–154, 2009.
Management of WNS in bats is a complex challenge similar 3. Lorch JM, Meteyer CU, Behr MJ, et al: Experimental infection
to other diseases in free-ranging wildlife populations. of bats with Geomyces destructans causes white-nose syndrome,
Current efforts are focused on integrated approaches that Nature 480:376–378, 2011.
reduce infection and mortality in bats, as well as promoting 4. Minnis AM, Lindner DL: Phylogenetic evaluation of Geomyces
and allies reveals no close relatives of Pseudogymnoascus destruc-
overall health of bat populations to support resistance to
tans, comb. nov., in bat hibernacula of eastern North America,
and recovery from WNS.44 Because WNS is not the only Fungal Biol 117:638–649, 2013.
cause of bat mortality and population decline,45,46 recovery 5. Langwig KE, Frick WF, Hoyt JR, et al: Drivers of variation in
and conservation of bat populations will require a holistic species impacts for a multi-host fungal disease of bats, Philos
management approach.47 Trans R Soc Lond, B, Biol Sci 371, 2016.
Supportive care for bats with WNS consists of providing 6. Turner GG, Reeder DM, Coleman JC: A five-year assessment
warmth, food, and water, which leads to full recovery within of mortality and geographic spread of white-nose syndrome in
512 SE C T I O N 14 Small Mammals
North American bats and a look to the future, Bat Research News 23. Frank CL, Michalski A, McDonough AA, et al: The resistance
52:13–27, 2011. of a North American bat species (Eptesicus fuscus) to white-nose
7. Frick WF, Puechmaille SJ, Hoyt JR, et al: Disease alters macro- syndrome (WNS), PLoS ONE 9:e113958, 2014.
ecological patterns of North American bats, Glob Ecol Biogeogr 24. Bernard RF, McCracken GF: Winter behavior of bats and the
24, 2015. progression of white-nose syndrome in the southeastern United
8. Russell RE, Thogmartin WE, Erickson RA, et al: Estimating the States, Ecol Evol 7:1487–1496, 2017.
short-term recovery potential of little brown bats in the eastern 25. Cryan PM, Meteyer CU, Boyles JG, et al: Wing pathology of
United States in the face of white-nose syndrome, Ecol Modell white-nose syndrome in bats suggests life-threatening disruption
314:111–117, 2015. of physiology, BMC Biol 8:135, 2010.
8a. Wibbelt G, Puechmaille SJ, Ohlendorf B, et al: Skin lesions 26. Reeder DM, Frank CL, Turner GG, et al: Frequent arousal from
in European hibernating bats associated with Geomyces destruc- hibernation linked to severity of infection and mortality in bats
tans, the etiologic agent of white-nose syndrome, PLOS ONE with white-nose syndrome, PLoS ONE 7, 2012.
8:e74105, 2013. 27. Warnecke L, Turner JM, Bollinger TK, et al: Inoculation of bats
9. Puechmaille SJ, Wibbelt G, Korn V, et al: Pan-European distri- with European Geomyces destructans supports the novel pathogen
bution of white-nose syndrome fungus (Geomyces destructans) not hypothesis for the origin of white-nose syndrome, Proc Natl Acad
associated with mass mortality, PLoS ONE 6, 2011. Sci USA 109:6999–7003, 2012.
10. Leopardi S, Blake D, Puechmaille SJ: White-nose syndrome 28. Warnecke L, Turner JM, Bollinger TK, et al: Pathophysiology of
fungus introduced from Europe to North America, Curr Biol white-nose syndrome in bats: a mechanistic model linking wing
25:R217–R219, 2015. damage to mortality, Biol Lett 9:20130177, 2013.
11. Hoyt JR, Sun K, Parise KL, et al: Widespread bat white-nose 29. Verant ML, Meteyer CU, Speakman JR, et al: White-nose
syndrome fungus, northeastern China, Emerging Infect Dis syndrome initiates a cascade of physiologic disturbances in the
22:140–142, 2016. hibernating bat host, BMC Physiol 14, 2014.
12. Lorch JM, Lindner DL, Gargas A, et al: A culture-based survey of 30. Cryan PM, Meteyer CU, Blehert DS, et al: Electrolyte depletion
fungi in soil from bat hibernacula in the eastern United States and in white-nose syndrome bats, J Wildl Dis 49:398–402, 2013.
its implications for detection of Geomyces destructans, the causal 31. Carr JA, Bernard RF, Stiver WH: Unusual bat behavior during
agent of bat white-nose syndrome, Mycologia 105:237–252, winter in Great Smoky Mountains National Park, Southeast Nat
2013. 13:N18–N21, 2014.
13. Zhang T, Victor TR, Rajkumar SS, et al: Mycobiome of the bat 32. Lorch JM, Minnis AM, Meteyer CU, et al: The fungus Tricho-
white nose syndrome affected caves and mines reveals diversity phyton redellii causes skin infections that resemble white-nose
of fungi and local adaptation by the fungal pathogen Pseudogym- syndrome of hibernating bats, J Wildl Dis 51:36–47, 2015.
noascus (Geomyces) destructans, PLoS ONE 9:e108714, 2014. 33. Janicki AF, Frick WF, Kilpatrick AM, et al: Efficacy of visual
14. Verant ML, Boyles JG, Waldrep W, Jr, et al: Temperature- surveys for white-nose syndrome at bat hibernacula, PLoS ONE
dependent growth of Geomyces destructans, the fungus that causes 10:e0133390, 2015.
bat white-nose syndrome, PLoS ONE 7:e46280, 2012. 34. Turner GG, Meteyer CU, Barton H, et al: Nonlethal screening
15. Brack V, Jr: Temperatures and locations used by hibernating of bat-wing skin with the use of ultraviolet fluorescence to detect
bats, including Myotis sodalis (Indiana bat), in a limestone mine: lesions indicative of white-nose syndrome, J Wildl Dis 50, 2014.
implications for conservation and management, Environ Manage 35. Meteyer CU, Valent M, Kashmer J, et al: Recovery of little
40:739–746, 2007. brown bats (Myotis lucifugus) from natural infection with Geo-
16. Geiser F: Metabolic rate and body temperature reduction during myces destructans, white-nose syndrome, J Wildl Dis 47:618–626,
hibernation and daily torpor, Annu Rev Physiol 66:239–274, 2011.
2004. 36. Fuller NW, Reichard JD, Nabhan ML, et al: Free-ranging little
17. Meteyer CU, Buckles EL, Blehert DS, et al: Histopathologic brown myotis (Myotis lucifugus) heal from wing damage associ-
criteria to confirm white-nose syndrome in bats, J Vet Diagn ated with white-nose syndrome, Ecohealth 8:154–162, 2011.
Invest 21:411–414, 2009. 37. Vanderwolf KJ, Malloch D, McAlpine DF: Detecting viable
18. Moore MS, Reichard JD, Murtha TD, et al: Specific alterations Psuedogymnoascus destructans (Ascomycota: Psueduerotiaceae)
in complement protein activity of little brown myotis (Myotis from walls of bat hibernacula: effect of culture media, J Caves
lucifugus) hibernating in white-nose syndrome affected sites, Karst Stud 78:158–162, 2016.
PLoS ONE 6:e27430, 2011. 38. Verant ML, Minnis AM, Lindner DL, et al: Geomyces and Pseu-
19. Field K, Johnson J, Lilley T, et al: The white-nose syndrome dogymnoascus: Emergence of a primary pathogen, the causative
transcriptome: activation of anti-fungal host responses in wing agent of bat white-nose syndrome: Chapter 28. In The fungal
tissue of hibernating bats, PLoS Pathog 11, 2015. community: its organization and role in the ecosystem, Boca Raton,
20. Johnson JS, Reeder DM, Lilley TM, et al: Antibodies to Pseu- FL, 2017, CRC Press, pp 405–415.
dogymnoascus destructans are not sufficient for protection against 39. Langwig KE, Frick WF, Reynolds R, et al: Host and pathogen
white-nose syndrome, Ecol Evol 5, 2015. ecology drive the seasonal dynamics of a fungal disease, white-
21. Meteyer CU, Barber D, Mandl JN: Pathology in euthermic nose syndrome, Proc Biol Sci 282:20142335, 2015.
bats with white-nose syndrome suggests a natural manifestation 40. Turner GG, Meteyer CU, Barton H, et al: Nonlethal screen-
of immune reconstitution inflammatory syndrome, Virulence 3, ing of bat-wing skin with the use of ultraviolet fluorescence to
2012. detect lesions indicative of white-nose syndrome, J Wildl Dis
22. Moosman PR, Veilleux JP, Pelton GW, et al: Changes in capture 50:566–573, 2014.
rates in a community of bats in New Hampshire during the 41. Reichard JD, Kunz TH: White-nose syndrome inflicts lasting
progression of white-nose syndrome, Northeast Nat (Steuben) injuries to the wings of little brown myotis (Myotis lucifugus),
20:552–558, 2013. Acta Chiropt 11:457–464, 2009.
CHAPTER 72 White-Nose Syndrome: Cutaneous Invasive Ascomycosis in Hibernating Bats 513
42. Powers LE, Hofmann JE, Mengelkoch J, et al: Temporal varia- 47. Maslo B, Valent M, Gumbs JF, et al: Conservation implications
tion in bat wing damage in the absence of white-nose syndrome, of ameliorating survival of little brown bats with white-nose
J Wildl Dis 49:946–954, 2013. syndrome, Ecol Appl 25:1832–1840, 2015.
42a. Ballmann AE, Torkelson MR, Bohuski EA, et al: Dispersal 48. Reichard JD, Fuller NW, Bennett AB, et al: Interannual survival
hazards of Pseudogymnoascus destructans by bats and human of Myotis lucifugus (Chiroptera: Vespertilionidae) near the epicen-
activity at hibernacula in summer, J Wildl Dis 53:725–735, 2017. ter of white-nose syndrome, Northeast Nat (Steuben) 21, 2014.
42b. Verant ML, Bohuski EA, Richgels KLD, et al: Determinants 49. Cheng TL, Mayberry H, McGuire LP, et al: Efficacy of a pro-
of Pseudogymnoascus destructans within bat hibernacula: Implica- biotic bacterium to treat bats affected by the disease white-nose
tions for surveillance and management of white-nose syndrome. syndrome, J Appl Ecol n/a–n/a, 2016.
J Appl Ecol 2018. 50. Cornelison C, Keel M, Gabriel K, et al: A preliminary report
43. Lorch JM, Muller LK, Russell RE, et al: Distribution and on the contact-independent antagonism of Pseudogymnoascus
environmental persistence of the causative agent of white-nose destructans by Rhodococcus rhodochrous strain DAP96253, BMC
syndrome, Geomyces destructans, in bat hibernacula of the Eastern Microbiol 14:1–7, 2014.
United States, Appl Environ Microbiol 79:1293–1301, 2013. 50a. Palmer JM, Drees KP, Foster JT, et al: Extreme sensitivity
44. Langwig KE, Voyles J, Wilber MQ, et al: Context-dependent to ultraviolet light in the fungal pathogen causing white-nose
conservation responses to emerging wildlife diseases, Front Ecol syndrome of bats, Nature Communications 9:35, 2018. https://
Environ 13:195–202, 2015. www.nature.com/articles/s41467-017-02441-z.
45. O’Shea TJ, Cryan PM, Hayman DTS, et al: Multiple mortality 51. Wilcox A, Willis CKR: Energetic benefits of enhanced summer
events in bats: a global review, Mammal Review 46:175–190, roosting habitat for little brown bats (Myotis lucifugus) recov-
2016. ering from white-nose syndrome, Conserv Physiol 4:cov070,
46. Ingersoll TE, Sewall BJ, Amelon SK: Effects of white-nose 2016.
syndrome on regional population patterns of 3 hibernating bat
species, Conserv Biol 30:1048–1059, 2016.
73
Naked Mole Rat Management
and Medicine
JAN RAINES
514
CHAPTER 73 Naked Mole Rat Management and Medicine 515
• Figure 73.1 Naked mole rat Heterocephalus glaber colony. • Figure 73.4 Chamber, again demonstrating the naturalistic interior;
that can become active within 24 hours of the loss of the • BOX 73.1 Techniques for Reintroducing Naked
queen. The reproductive activation of several females and Mole Rats (Heterocephalus glaber)
increases in intracolony aggression, social instability, and into Colony Post Procedure
body weight often follow a queen’s death. Females older
than 6 months are capable of becoming reproductively • Attempt to wear exam gloves at all times while handling
animals to reduce scent transmission.
active and fight to establish dominance if a queen dies. • Do not perform surgical prep with fragrance-containing items.
The death of colony members following queen removal • If an animal does not have a transponder, temporarily identify
may be an extreme attempt by new queens to establish with a marker for any follow-up monitoring.
their dominance and reimpose reproductive suppression on • Remove multiple animals from the colony, and reintroduce
subordinates. During these upheavals, both sexes sustain the lone animal to that group before reintroducing to the
entire group.
serious injuries as they battle to establish dominance and • The maximum time out of colony should be no more than 2
become breeding animals.6,19–22 hours.
In captive colonies, females have produced litters as large • Have soiled shavings available so that the animal may be
as 27, whereas it is reported that litter size is closer to 12 scented with them prior to returning to the colony.
pups in wild populations. Because NMRs are year-round
breeders, captive populations may reach carrying capacity
very rapidly. In wild colonies, dispersal is a valid option to
reduce population size or reduce incidence of inbreeding,
with the animals merely sealing off a new colony in a
group of tunnels to divide them from the originating group.
When a colony splits into two groups, one of the non-
breeding females in the new colony will ascend to become
queen.14,23,24 There is a strong indication that rapid colony
growth in limited space contributes to high pup mortality
and is correlated to the size of the breeding colony.13 Colony
collapse occurs when the colony has exceeded carrying
capacity, and pup mortality is high (95%–100%).
In captivity, where there is limited space for the colony
to expand, finding a successful method of contraception
without causing queen succession has been challenging.
Three successful methods have been used to date: surgi-
cal sterilization, hormonal manipulation, and vaccination
(see also Chapter 22). A tubal ligation and a complete • Figure 73.6 Naked mole rat (Heterocephalus glaber) under fenes-
hysterectomy are successful methods of surgical steriliza- trated drape with melengestrol acetate implant indicated at point of
tion, but an ovariohysterectomy is not an option because scalpel blade.
removal of ovarian tissue would cause the queen to lose her
status and instigate a succession coup. Not knowing how a second queen was implanted with melengestrol acetate
well an individual could be reintroduced after an invasive (MGA) (Fig. 73.6). A concern with the use of MGA was
surgical procedure, test procedures were performed on other the suppression of the pheromones maintaining sterility
females in the colony. NMRs are highly xenophobic, and because its use could stimulate a fight for dominance. MGA
they will kill individuals that they do not recognize, as well is a synthetic progestin affecting contraception by blocking
as animals from their own colony that have been removed ovulation and thickening cervical mucus; however, follicle
for an excessive length of time.25 Aggression post removal growth and estrous behavior continue. An MGA implant
can occur in as brief a timeframe as 30 minutes or not (0.15 mg/kg, ZooPharm, Windsor, Colorado 80550) was
until almost 2 hours; some of this range is dependent on placed in a routine fashion, and reproduction was curtailed
the individual’s social standing when removed. Preventative for more than 2 years, but implant failure occurred at 26
measures may be used to reduce the risk of injury whenever months with confirmed pregnancy. A queen in another
an animal is removed from a colony even for a short period colony received a porcine zona pellucida (PZP) vaccine
of time (Box 73.1). A flank hysterectomy where the uterus in an attempt to curtail reproduction. The PZP vaccine
was removed from a single incision sparing the ovaries was causes antibodies that prevent attachment of sperm to the
accomplished in a subordinate female who was introduced ova. It does not appear to suppress estrous cycle, and it
to the colony without incident following the procedure. The is easy to administer. A dosing regimen of 0.2 mL PZP
tubal ligation surgery has also been performed via both a emulsion (50 µg) intraperitoneally was administered. The
flank approach and ventral midline, again with successful queen received one injection followed by a booster injection
return to the colony. Flank approaches are recommended to 2 weeks later. With the initial series and an annual booster
reduce incidence of visceral herniation if colony members schedule, there have been no pups to date (at time of
chew at surgical sites. As an alternative sterilization method, publication, more than 3 years).
CHAPTER 73 Naked Mole Rat Management and Medicine 517
Therapeutics
Many oral antibiotics (TMPS at 15–30 mg/kg orally [PO]
once or twice a day [SID or BID] and enrofloxacin at 5 mg/
• Figure 73.7 Naked mole rat (Heterocephalus glaber) with calcinosis
kg PO or intramuscularly [IM] SID) and nonsteroidal cutis/circuscripta. (From Delaney MA, Nagy L, Kinsel MJ, Treating PM:
antiinflammatory drugs (meloxicam at 0.1–0.2 mg/kg PO Spontaneous histologic lesions of the adult naked mole rat [Hetero-
SID) at standard rodent dosing regimens have been used cephalus glaber] a retrospective survey of lesions in a zoo population,
successfully in this species, with administration volume Vet Path 50[4]:607–621, 2013.)
being the only limiting factor. Response to analgesics may
be minimal in this species because they lack C-fibers in
addition to substance P and nerve growth factor, both neu- and they maintain fertility until death.28–31 NMRs also have
rotransmitters in pain cascades, which decrease or eliminate a higher expression of cytoprotective genes allowing them
any cutaneous or thermal pain sensation. This adaptation is to resist overt disease.8,26
thought to be part of a protective mechanism to reduce acid
accumulation in tissues in their hypoxic environment.8,26,27
References
Diseases (Infectious and Noninfectious) 1. Jarvus JUM: Eusociality in a mammal: cooperative breeding in
naked mole-rat colonies, Science 212(4494):571–573, 1981.
The most commonly reported disease conditions in NMRs 2. Bennett NC, Jarvis JUM: The social structure and reproduc-
are related to trauma secondary to intraspecific aggression. tive biology of colonies of the mole-rat Cryptomys damarensis
The most common bacteria isolated from bite wound (Rodentia: Bathyergidae), J Mammal 69:293–302, 1988.
abscesses have been Pasturella sp. Noninfectious disease 3. Larson J, Park TJ: Extreme hypoxia tolerance of naked mole-rat
issues are most often directly related to husbandry. Neo- brain, Neuroreport 20(18):1634–1637, 2009.
natal death from maternal neglect is most often seen with 4. Maina JN, Gebreegziabher Y, Woodley R, et al: Effects of change
impending colony collapse. Neglected animals are dead in environmental temperature and natural shifts in carbon dioxide
within 12 hours of birth, and they are often moved by the and oxygen concentrations on the lungs of captive naked mole-
worker animals into the “potty” chamber shortly after, and rats (Heterocephalus glaber): a morphological and morphometric
study, J Zoo 253:371–382, 2001.
sometimes before, death. Calcinosis circumscripta and cutis
5. Honeycutt RL: Naked mole rats, Am Sci 80:43–53, 1992.
has been reported in multiple colonies. The animals present 6. Nowak RM: Rodentia. In Walker’s mammals of the world, ed 6,
with subcutaneous swellings that range in size from 2 mm Baltimore, MD, 1999, The Johns Hopkins University Press, pp
to 2 cm and contain a white, opaque, pasty material, and 1642–1644.
these lesions can be of such size that the animal is incapable 7. Buffenstein R, Jarvis JUM: The naked mole rat: a new record for
of movement (Fig. 73.7). This condition is attributed to the oldest living rodent, Sci Aging Knowl Environ 2002(21):4,
inappropriate vitamin D supplementation within the diet; 2002.
however, even after vitamin D supplementation was discon- 8. Delaney MA, Nagy L, Kinsel MJ, et al: Spontaneous histologic
tinued, it was reported that the pups from the subsequent lesions of the adult naked mole rat (Heterocephalus glaber): A
litters born immediately around the time of discontinuation retrospective survey of lesions in a zoo population, Vet Path
matured to develop these lesions. 50(4):607–621, 2013.
9. Artwhol MS, Hill T, Corner C, et al: Naked mole-rats: unique
opportunities and husbandry challenges, Lab Anim 31(5):32–36,
Use in Research 2002.
10. Buffenstein R, Yahav S: The effect of diet on microfaunal
Because of their unique biology and physiology, NMRs are population and function in the caecum of a subterranean naked
valuable models for aging research. NMRs age at a slower mole-rat, Heterocephalus glaber, Br J Nutr 65(2):249–258, 1991.
rate; they have increased resilience to oxidative stress and 11. Buffenstein R, Laundy MT, Pitcher T, et al: Vitamin D3
mitochondrial injury; they have minimal cellular senescence; intoxication in naked mole-rats (Heterocephalus glaber) leads to
518 SE C T I O N 14 Small Mammals
hypercalcaemia and increased calcium deposition in teeth with 21. Margulis SW, Saltzman W, Abbott DH: Behavioral and hor-
evidence of abnormal skin calcification, Gen Compar Endocrinol monal changes in female naked mole-rats (Heterocephalus glaber)
99(1):35–40, 1995. following removal of the breeding female from a colony, Horm
12. Pitcher T, Buffenstein R: Intestinal calcium transport in mole- Behav 29:227–247, 1995.
rats (Cryptomys damarensis and Heterocephalus glaber) is indepen- 22. Goldman SL, Forger NG, Goldman BD: Influence of gonadal
dent of both genomic and non-genomic vitamin D mediation, sex hormones on behavioral components of the reproductive
Experim Physiol 80:597–608, 1995. hierarchy in naked mole-rats, Horm Behav 50:77–84, 2006.
13. Roellig K, Drews B, Goeritz F, et al: The long gestation of the small 23. Ciszek D: New colony formation in the “highly inbred” eusocial
naked mole-rat (Heterocephalus glaber RÜPPELL, 1842) Studied naked mole-rat: outbreeding is preferred, Behav Ecol 11:1–6,
with Ultrasound biomicroscopy and 3D-Ultrasonography, PLoS 1999.
ONE 6:1–10, 2011. 24. Braude S: Dispersal and new colony formation in wild naked
14. Faulkes CG, Abbott DH, Jarvis JUM: Social suppression of mole-rats: evidence against inbreeding as the system of mating,
ovarian cyclicity in captive and wild colonies of naked mole-rats, Behav Ecol 11:7–12, 1999.
Heterocephalus glaber 88:559–568, 1990. 25. Clarke FM, Faulkes CG: Intra-colony aggression in the naked
15. Clarke FM, Faulkes CG: Hormonal and behavioural correlates mole-rat, Heterocephalus glaber, Anim Behav 61:311–324,
of male dominance and reproductive status in captive colonies 2001.
of the naked mole-rat, Heterocephalus glaber, Proc Biol Sci 26. Omerbasic D, Smith ESJ, Moroni M, et al: Hypofunctional
264(1404):1391–1399, 1998. TrkA accounts for the absence of pain sensitization in the African
16. Faulkes CG, Abbott DH, Jarvis JUM: Social suppression of naked mole-rat, Cell Rep 17:748–758, 2016.
reproduction in male naked mole-rats, Heterocephalus glaber, J 27. Park TJ, Lu Y, Juttner R, et al: Selective inflammatory pain
Reprod Fertil 91:593–604, 1991. insensitivity in the African naked mole-rat (Heterocephalus
17. Faulkes CG, Trowell SN, Jarvis JU, et al: Investigation of glaber), PLoS Biol 6(1):e13, 2008.
numbers and motility of spermatozoa in reproductively active 28. Andziak B, Buffenstein R: Disparate patterns of age-related
and socially suppressed males of two eusocial African mole-rats, changes in lipid peroxidation in long-lived naked mole-rats and
the naked mole-rat (Heterocephalus glaber) and the Damaraland shorter-lived mice, Aging Cell 5(6):525–532, 2006.
mole-rat (Cryptomys damarensis), J Reprod Fertil 100:411–416, 29. Andziak B, O’Connor TP, Qi W, et al: High oxidative damage
1994. levels in the longest-living rodent, the naked mole-rat, Aging Cell
18. Onyango DW, Oduor-Okelo D, Otiang’a-Owiti GE: Ultra- 5(6):463–471, 2006.
structural study of the testes of non-breeding naked mole-rat 30. Buffenstein R: The naked mole-rat: A new long-living model
(Heterocephalus glaber), Ann Anat 175(5):447–452, 1993. for human aging research, J Gerontol Bio Sci 60(11):1369–1377,
19. Clarke FM, Faulkes CG: Dominance and queen succession in 2005.
captive colonies of the eusocial naked mole-rat, Heterocephalus 31. Buffenstein R: Negligible senescence in the longest living rodent,
glaber, Proc Biol Sci 264:993–1000, 1997. the naked mole-rat: insights from a successfully aging species, J
20. Faulkes CG, Abbott DH: Social control of reproduction in Compar Physiol B 178(4):439–445, 2008.
breeding and non-breeding male naked mole rats (Heterocephalus
glaber), J Reprod Fert 93:427–435, 1991.
74
Immobilization, Health, and
Current Status of Knowledge
of Free-Living Capybaras
SANTIAGO MONSALVE
Introduction
Ecuador, Venezuela, Brasil, Perú, Bolivia, and the Guyanas,
This chapter is based on work done in the Colombian while the second inhabits northern Colombia, Panama,
Caribbean and Orinoquía region in which investigative and Venezuela.6 The rapid decrease of wild populations
processes have been undertaken in the area of conserva- in Colombia for decades has led to the taking of diverse
tion medicine with capybaras (Hydrochoerus spp.) in in situ measures to ensure their conservation, while at the same
conditions. time satisfying the interests of the population for exploiting
this resource in the different countries in which they are
The Capybara naturally found in situ.
The capybara is the largest living rodent species in the world Habitat
and the only surviving member of the family Hydrochoeri-
dae. This species is found in a geographic distribution area The transformation of natural savannahs since the time of
that includes Panamá, Colombia, Venezuela, Ecuador, Perú, colonization and the expansion of cattle ranching have been
Paraguay, Uruguay, Brazil, Bolivia, and northern Argen- threats for its habitat. Petroleum exploration and exploitation,
tina.1 Its semiaquatic behavior is an advantage due to the the incursions of roads, and the presence of groups on the
fact that it can procreate in areas that are subject to marginal margins of the law have also generated significant processes
hydrologic fluctuations, cattle ranching, and agriculture. Its of transformation of the ecosystem due to environmental
condition as a native species, adapted to swampy areas and impacts.7 Likewise, the expansion of monocultures has been
areas subject to flooding, in contrast to bovine and equine another factor that has affected the habitat of capybaras
domesticated species, provides it with greater efficiency in and constitutes a threat to their populations, as this activity
the utilization of available forage in these environments.2 implies the use of agrochemicals that deteriorate the quality
The capybara is a species of great economic interest, due of water, which is an important resource for this species.
to its high fecundity and the excellent quality of its meat In Colombian Orinoquía and Venezuela, great swaths of
and skin, characterized by the fact that countries such as savannah flood during the rainy season, increasing available
Brazil, Venezuela, and Argentina have established successful habitat for animals; nevertheless, in some areas, flooding
programs for management of the species.3–5 may be extreme and these floodwaters may be considered
to be a limiting factor, because these animals also require
Geographic Distribution dry zones to rest. During summer, these bodies of water
shrink to the point that they completely disappear; at this
The capybara, also known as the chigüiro, ponche, capy- moment, the available and optimal habitat for these animals
ibara, cacó, and carpincho, belongs to the order Rodentia, is reduced to small strips around remaining water sources
genus Hydrochoerus. The capybara is autochthonous to (Fig. 74.1).8 In actuality, thousands of capybaras die during
the American continent, where there are two species: the dry season without scientists having clarity about the
Hydrochoerus hydrochaeris and Hydrochoerus isthmius, the population equilibrium that this anthropic condition may
first with a distribution in the Llanos Orientales and Ama- be generating or the epidemiologic implications associated
zonian region of Colombia, northern Argentina, Paraguay, with this annual mortality.
519
520 SE C T I O N 14 Small Mammals
Biology and State of Conservation may be found in habitats close to bodies of water, rivers,
and streams,3 and can be considered to be the most efficient
Despite capybaras being rodents and having high reproduc- nitrogen recyclers of all animals, as, in a matter of hours,
tive rates, the fear exists that legal and illegal extraction may their urine makes important quantities of nitrogen soluble,
prevent this species from maintaining sustainable popula- which are re-integrated into pastures.16 Sufficient data for
tions over time.9,10 Capybaras are highly social and their the cataloguing of the species H. isthmius do not exist,
population dynamics are intensely related to reproductive according to the IUCN.6
competition in males and females.11 Demographic changes
in capybara populations can be modified due to differ- Capybara as Microorganism Amplifier
ent social mechanisms and habitual infanticide in which
groups of male individuals may kill juvenile competitors Diseases transmitted by vectors are not circumscribed to
in order to assume population control.12,13 This behavior definite regions and have been recognized in practically any
has been considered to be an ancestral trait in rodents13,14 place they have been investigated.17 The risks of the major-
and an important estrual inductor in females, reducing the ity of diseases contracted from wild animal populations
period of time between litters.12,13 This species counts on are unknown, as is their impact. Many of these diseases
reproductive suppression on the part of females,15 which may eventually affect man,21 increasing epidemiologic
may have several young in the group; however, not neces- alterations, especially when a species such as the capybara
sarily all the females of the group are reproductively active is utilized for the consumption of its meat, because it is a
during certain periods of time, which suggests mechanisms species that cohabitates with domestic animals and humans
of estrual suppression.13,15 According to the International in large areas immersed in extensive agricultural production.
Union for Conservation of Nature (IUCN), the species Capybaras have been reported to be transmitters of diseases,
H. hydrochaeris is catalogued as a Least Concern species and it has been determined that populations of this species
due to its wide geographic distribution, high and prolific in the Casanare Department (Colombia) have or have had
population, and permanence in protected areas.6 Capybaras contact with Brucella abortus and Leptospira interrogans.22
Populations of capybaras have been diminished during dry
seasons, and combined with greater human intervention in
the environment, as is presented in this department, may
represent an indicator of increase in density of disease-
causing organisms.23,24
It has been determined that horses, cattle, tapirs (Tapirus
terrestris), and capybaras (Hydrochoerus spp.) are infected
by ticks with the Amblyomma cajennense complex, as well
as domestic animals and humans.25 This genus of ticks has
been considered to be a vector of diverse microorganisms;
thus the capybara has been catalogued as an animal ampli-
• Figure 74.1 Wild capybara (Hydrochoerus hydrochaeris) habitat. fier of rickettsial diseases, as it fits certain requirements that
Casanare, Colombia. permit disease cycles within its habitat (Fig. 74.2).26,27
Abundance of
Detection of large
capybaras in their
periods of bacteremia
natural habitat
• Figure 74.2 Characteristics necessary for an organism to be considered to be a reservoir for the
amplification of Rickettsias.
CHAPTER 74 Immobilization, Health, and Current Status of Knowledge of Free-Living Capybaras 521
A B
• Figure 74.3 (A) Amblyomma spp. and (B) Dermacentor spp. obtained from a capybara (Hydrochoerus
hydrochaeris).
Different studies have demonstrated that capybaras are urgently require a constant epidemiologic vigilance; thus
amplifying hosts of Rickettsia rickettsii (the microorganism due to the fact that capybaras are considered to be amplify-
that causes Rickettsiosis, a zoonotic disease with a high ing hosts of diverse microorganisms,27,28 it is important to
percentage of lethality) and are commonly parasitized by take into consideration the diagnosis of zoonotic diseases of
ticks of the A. cajennense complex, a principal vector of rickettsial origin, including the microorganisms Rickettsia
rickettsiosis in South America (Fig. 74.3A)27; however, they spp., Ehrlichia spp., and Anaplasma spp., as pathogens that
have also been shown to be infested by other species of represent a risk in the transmission of human and animal
ectoparasites such as ticks of the genus Dermacentor spp. diseases.
(see Fig. 74.3B). The increase in cases of maculous fever
in Brazil is related to the presence of capybaras in the area
due to their condition as primary host amplifiers of these Immobilization
microorganisms.28 Physical Immobilization
As they are considered to be susceptible to R. rickettsii
and by being primary hosts of A. cajennense, capybaras may Two methods may be used to physically capture wild
be epidemiologic bioindicators of Rickettsia infections in capybaras.
specific geographic areas.
The ticks that infest wild capybaras simultaneously trans- Capture Corrals
mit different microorganisms to individuals of the species, The purpose of this strategy is to condition capybaras during
to other wild and domestic animals, and to humans. In the space of 1–3 months, generally during the dry season,
studies undertaken in two states of Brazil (Rondonia and Sao to approach a corral by baiting it every day with food.
Paulo), the DNA of Ehrlichia spp. has not been detected by These corrals have a guillotine style door that is supported
molecular techniques in the amplification of fragments of at a distance by means of a string that acts as a latch and
gen dsb29; in Colombia (Casanare Department) the circula- remains open, permitting access to the interior of the corral
tion of microorganisms of the family Anaplasmataceae have so that they can be captured for investigative means. In
been found using polymerase chain reaction (PCR) for the studies done in the Colombian Caribbean region for the
amplification of a segment of gen 16S rRNA por tqPCR.30 process of conditioning, rations of sugarcane, carrots, ears
Preliminary studies on ticks of the A. cajennense complex of corn, and cut grass have been used, which were moved
have shown identities similar to Ehrlichia spp. and Ana- incrementally each day from the wetlands that the capybaras
plasma spp. The results of these studies have permitted inhabit toward the interior of the corral. The capture corrals
the gleaning of preliminary evidence of the circulation of are constructed piece by piece so that the animals gradually
pathogens to the family Anaplasmataceae of capybaras in become accustomed to the structure. Although it is a rela-
in situ conditions.31 tively costly and time-intensive methodology, it represents a
Although in the last century there was important pro- secure methodology for the animals and operators that want
gress made in the control of infectious diseases, these still to utilize corral capture.2 This technique is very useful next
represent an important problem for public health, princi- to lakes and rivers and may yield the capture of between 10
pally in terms of zoonotic microorganisms that continue to and 20 capybaras per month.
522 SE C T I O N 14 Small Mammals
Conclusions
The capybara is a wild animal that is resistant to disease
and adapts well to rural environments, making it an excel-
• Figure 74.5 Chemical immobilization of a free-living capybara
(Hydrochoerus hydrochaeris).
lent agricultural alternative (subsistence consumption);
however, the use of the species for this purpose presents
various challenges, as its habitat changes seasonally. The
Roping high variability in the behavior of this forager allows the
This methodology requires the assistance of cowboys in the species to adapt to changes in feeding, which explains
geographic zones in which physical restraint is to proceed. the high plasticity of capybaras in dealing with environmen-
Immobilization is done by chasing the animals over the tal changes and permits them to colonize different types
savannah on horseback, during which, generally, the of habitats. Due to its proximity to human populations
cowboys use lariats for restraint (Fig. 74.4). This technique and consumption of wildlife, the capybara is a species that
is very useful in large, flat zones and may yield the capture could be of great importance to the monitoring of emerging
of between 5 and 10 capybaras per day. The cost of this infectious diseases and the potential reemergence of zoonot-
methodology may be less than physical capture using corrals ics as ecosystemic bioindicators. Deeper investigations into
and may yield proportionally a larger number of animals the distribution and ecology of wild reservoirs of these
per day. pathogens and their vectors are required.
Andrenergic Tiletamine
Geographic Ketamine Agonists (α2) Zolacepam
No. Region Samples Dose Dose Dose Others Antagonists Reference Comments
1 Colombian 8 — — 5 mg/kg — Not reported 2 Safe anesthetic combination. In
Caribbean higher doses, may have been
region used to induce anesthesia for a
larger period of time.
2 Colombian 8 10 mg/kg Xylacine — — Not reported 2 Safe anesthetic combination. May
Caribbean 0.5 mg/kg produce some brachycardia
region and inspiratory dyspnea.
3 State of Minas 10 10 mg/kg Xylacine — — Not reported 33 Best analgesic in integument.
Gerais, Brazil 0.5 mg/kg
4 State of Minas 10 — — 5 mg/kg — Not reported 33 Long-duration anesthesia with
Gerais, Brazil low analgesia.
5 State of Minas 10 — — 5 mg/kg Levomepromazine Not reported 33 Safe anesthetic combination,
Gerais, Brazil 0.5 mg/kg good analgesic
6 State of Apure, 16 4 mg/kg — — Fenotiazine 0.4 mg/kg Not reported 35 Good immobilization without
Venezuela inducing total anesthesia
7 State of Apure, 12 4 mg/kg Medetomidine — — Atipamezole 35 Good immobilization without
Venezuela 0.04 mg/kg 0.1 mg/kg inducing total anesthesia, risk
of postanesthesia suffocation
8 Curitiba, Brazil 22 — — 3 mg/kg Morphine 0.3 mg/kg + Not reported 35 Sedation for opthamalic tests
Azaperone 1.2 mg/
kg
9 State of Apure, 8 — — 5.1 mg/kg — Not reported 36 Safe anesthetic combination, long
Venezuela duration
10 State of Apure, 6 — Medetomidine 1.6 mg/kg Not reported 36 Safe anesthetic combination
Venezuela 0.008 mg/kg
11 State of Apure, 8 — Medetomidine 1.5 mg/kg Butorfanol 0.075 Not reported 36 Safe anesthetic combination,
Venezuela 0.0075 mg/kg good analgesic
CHAPTER 74 Immobilization, Health, and Current Status of Knowledge of Free-Living Capybaras
523
524 SE C T I O N 14 Small Mammals
TABLE
74.2 Infectious Diseases Reported in the Capybara (Hydrochoerus spp.)
Disease/Reservoir/ Signs/Symptoms in
Classification Agent Zoonosis Capybaras Reference
Virals Picornaviridae Foot-and-mouth disease Vesicular interdigital lesions 39
Picornaviridae Viral Miocarditis necrotizante 40
encephalomyocarditis
Poxviridae Orthopoxvirus Zoonosis: fever, Asymptomatic 41
lymphadenopathy,
headache and malaise
Retroviridae BLV Not reported 42
Bacterial Rickettsia bellii, Rickettsia Spotted fever rickettsiosis Asymptomatic 43
parkeri
Escherichia coli O157 Zoonosis: hemorrhagic Not reported 44
enterocolitis. Disease not
reported in capybaras
Rickettsia rickettsi Amplifier: Rocky Mountain Asymptomatic 27, 45
spotted fever
Leptospira Reservoir of leptospirosis Not reported 46
Icterohaemorrhagiae, L.
grippotyphosa and L.
shermani
Brucella spp. Zoonosis: Brucelosis. Not reported 47, 48
Reservoir of Brucella
abortus
Ehrlichia spp. and Zoonosis: Human granulocytic Not reported 31
Anaplasma spp. anaplasmosis. In canines:
Anaplasmosis and
ehrlichiosis
Mycobacterium bovis, Tuberculosis Tubercular Granulomas, 49, 50
Mycobacterium glomerulonephritis, pancreatic
intracellulare fibrosis. Mesenteric
granulomatous nodules
Parasitic Fasciola hepatica Fasciolosis Abortions, infertility, hepatic 51–53
lesions
Protohallidae, pycnotrichidae, Unreported disease, Asymptomatic 54
cycloposthiidae families endosymbiont
Tripanosoma evansi Reservoir Asymptomatic 55, 56
Género Amblyomma spp. Amplifier of Rickettsial Asymptomatic 43, 57
microorganisms
Cryptosporidium parvum Zoonosis: Nonsanguinary May present acute sanguinary 48, 58
watery diarrhea, anorexia, diarrhea
vomiting
Monoecocestus hagmani Intestinal parasitism Weight loss, anemia, abdominal 48
distention
9. Federico P, Canziani G: Modeling the population dynamics a potential model of epidemiological alert in endemic areas,
of capybara Hydrochoerus hydrochaeris: a first step towards a Biomedica 31(2):216–221, 2011.
management plan, Ecol Modell 186:111–121, 2005. 28. De Souza C, Berger S, Camargo C: Papel da capivaras
10. Mesa E: Análisis de la dinámica de poblaciones silvestres de Hydrochoerus hydrochaeris na cadeia epidemiológica da febre
chigüiros Hydrochoerus hydrochaeris (Linnaeus, 1766) como maculosa brasileira, Rev Bras Parasitol Veterinária 13(1):203–205,
herramienta de manejo hacia el aprovechamiento sostenible y la 2004.
conservación. Universidad Nacional de Colombia, 2005. 29. Labruna MB, McBride JW, Camargo LMA, et al: A preliminary
11. Armitage K, Bhnnstem DT: Life history consequences of social investigation of Ehrlichia species in ticks, humans, dogs, and
complexity: a life history consequences of social complexity: a capybaras from Brazil, Vet Parasitol 143(2):189–195, 2007.
comparative study of ground-dwelling sciurids, 1998, 2014, 30. Zhang Q, Wang J, Deng F, et al: TqPCR: A touchdown qPCR
p 9. assay with significantly improved detection sensitivity and ampli-
12. Anthony LL, Blumstein DT: Integrating behaviour into wildlife fication efficiency of SYBR green qPCR, PLoS ONE 10(7):1–19,
conservation: the multiple ways that behaviour can reduce, 2000: 2015.
95. 31. Gómez M, González S, Rojano C, et al: Detección de Ehrlichia
13. Maldonado-Chaparro A, Blumstein DT: Management implica- spp y Anaplasma spp en chigüiros (Hydrochoerus hydrochaeris) de
tions of capybara (Hydrochoerus hydrochaeris) social behavior. vida silvestre y sus garrapatas en el departamento del Casanare,
2008; 1. Colombia. In Proceedings of the Primer congreso latinoameri-
14. Swenson JE: Implications of sexually selected infanticide for the cano de medicina y conservación de especies neotropicales, Cali,
hunting of large carnivores. In Festa-Bianchet M, Apollonio M, Colombia: ALVEFAS, 2016.
editors: Animal behavior and wildlife conservation, Washington, 32. Ferrari L, Turrini G, Rostello C, et al: Evaluation of two
DC, 2003, Island Press, pp 171–189. combinations of Domitor, Zoletil 100, and Euthatal to obtain
15. Solomon NG, French JA: The study of mammalian cooperative long-term nonrecovery anesthesia in Sprague-Dawley rats, Comp
breeding. In Solomon NG, French JA, editors: Cooperative breed- Med 55(3):256–264, 2005.
ing in mammals, 1997, Cambridge University Press, pp 1–10. 33. Nishiyama SM: Associação cetamina-xilazina, tiletamina-
16. Hume ID: Digestive strategies of mammals. Application of zolazepam, e tiletamina-zolazepam-levomepromazina na anestesia
chemical reactor theory to digestion, Acta Zool Sin 48:1–19, de capivara (Hydrochoerus hydrochaeris). Universidade Federal de
2002. Viçosa, 2003.
17. Da Silva LJ: Enfermedades transmitidas por garrapatas en 34. West G, Heard D, Caulkett N: Rodents. In Zoo animal and
humanos. Ocurrencia, distribución e impacto en salud pública, wildlife immobilization, ed 1, Ames, IA, 2007, Blackwell Profes-
con énfasis en el estado de Sao Paulo. Consult OPS/OMS Expert sional Publishing, p 655.
sobre rickettsiosis en las Américas Inf Final Bras. 2004. 35. Salas V, Pannier E, Galíndez-silva C, et al: Methods for captur-
18. Miranda J, Mattar S: Molecular detection of Rickettsia bellii ing and marking wild capybaras in Venezuela, Wildl Soc Bull
and Rickettsia sp. strain colombianensi in ticks from Cordoba, 32(1):202–208, 2004.
Colombia, Ticks Tick Borne Dis 5(2):208–212, 2014. 36. King JD, Congdon ÃE, Tosta C: Evaluation of three immobiliza-
19. Almeida AP, Cunha LM, Bello ACPP, et al: A novel Rickettsia tion combinations in the capybara (Hydrochoerus hydrochaeris),
infecting Amblyomma dubitatum ticks in Brazil, Ticks Tick Borne Zoo Biol 67:59–67, 2010.
Dis 2(4):209–212, 2011. 37. González Jiménez E: El capybara capybara capybara (Hydrochoe-
20. Monsalve S, Mattar S, González M: Silvestres y Su Impacto rus hydrochaeris). Estado actual de su producción. Direccion de
En Las Enfermedades Emergentes Y Reemergentes, Rev MVZ Produccion y Sanidad Animal FAO, Rome, Italy, 1995.
Córdoba 14(2):1762–1773, 2009. 38. Chiacchio RG, Eloisa F, Prioste S, et al: Health evaluation and
21. Acha PN, Szyfres B: Zoonoses and communicable diseases survey of zoonotic pathogens in free-ranging capybaras (Hydro-
common to man and animals. In: Parasitic Zoonoses. Pan choerus hydrochaeris), J Wildl Dis 50(3):496–504, 2014.
American Health Organization, 2003. 39. Gomes I, Rosenberg FJ: A possible role of capybaras (Hydrochoe-
22. Barragán K, Álvarez-Méndez O: Estudios serológicos de Brucella rus hydrochaeris hydrochoeris) in foot-and-mouth disease (FMD)
abortus y Leptospira interrogans en poblaciones silvestres de endemicity, Prev Vet Med 3(2):197–205, 1984.
chigüiros (Hydrochoerus hydrochaeris) en el departamento de 40. Wells SK, Gutter AE, Soike KF, et al: Encephalomyocarditis
Casanare. In El chigüiro Hydrochoerus hydrochaeris en la Orino- virus: epizootic in a zoological collection, J Zoo Wild Anim Med
quía Colomb Ecol manejo Sosten y Conserv, 2014, pp 197–210. 20(3):291–296, 1989.
23. Primack R, Rozzi R, Feinsinger P, et al: Especies exóticas, enfer- 41. Barbosa AV, Medaglia MLG, Soares HS, et al: Presence of neu-
medades y sobreexplotación. In Fundamentos de conservación tralizing antibodies to Orthopoxvirus in Capybaras (Hydrochoerus
biológica, México, 2001, Fondo de Cultura Económica, pp hydrochaeris) in Brazil, J Infect Dev Countries 8(12):1646–1649,
225–252. 2014.
24. Daszak P, Cunningham AA, Hyatt AD: Emerging infectious 42. Dimitrov PS, Todorova KS, Petrichev MH, et al: Bovine leu-
diseases of wildlife - threats to biodiversity and human health, kemia virus-pathogenicity in animals and potential impacts in
Science 287(January):443–449, 2000. humans, Cyprus J Sci 10:101, 2012.
25. Labruna MB: Ecology of rickettsia in South America, Ann N Y 43. Pacheco RC, Horta MC, Moraes-filho J, et al: Rickettsial infec-
Acad Sci 1166:156–166, 2009. tion in capybaras (Hydrochoerus hydrochaeris) from São Paulo,
26. Carlos J, Vélez Q, Hidalgo M, et al: Rickettsiosis, una enfer- Brazil: serological evidence for infection by Rickettsia bellii and
medad letal emergente y re-emergente en Colombia, Universitas Rickettsia parkeri, Biomedica 25:364–371, 2007.
Scientiarum 17:82–99, 2012. 44. Leotta GA, Deza N, Origlia J, et al: Detection and characteriza-
27. Miranda J, Contreras V, Negrete Y, et al: Surveillance of tion of Shiga toxin-producing Escherichia coli in captive non-
Rickettsia sp. infection in capybaras (Hydrochoerus hydrochaeris) domestic mammals, Vet Microbiol 118:151–157, 2006.
526 SE C T I O N 14 Small Mammals
45. Souza CE, Moraes-filho J, Ogrzewalska M, et al: Experimental e em bovinos (Bos taurus) no município de Timbó, Revista de
infection of capybaras (Hydrochoerus hydrochaeris) by Rickettsia Ciências Agroveterinárias 8(1):66–70, 2009.
rickettsii and evaluation of the transmission of the infection to 53. Dracz RM, Ribeiro VM, Pereira A, et al: Occurrence of
ticks (Amblyomma cajennense), Vet Parasitol 161:116–121, 2009. Fasciola hepatica (Linnaeus, 1758) in capybara (Hydrochoerus
46. Ferreira N, Albuquerque D, Martins G, et al: The role of capy- hydrochaeris) (Linnaeus, 1766) in Minas Gerais, Brazil, Rev Bras
baras as carriers of leptospires in periurban and rural areas in the Parasitol Veterinária 25(3):364–367, 2016.
western Amazon, Acta Trop 169:57–61, 2017. 54. Ito A: Ciliates from the cecum of capybara (Hydrochoerus hydro-
47. Lord VR, Ricardo FC, Lord R, et al: Brucella spp. from the chaeris) in Bolivia. The family Cycloposthiidae, Eur J Protistol
capybara (Hydrochoerus hydrochaeris) in Venezuela: serologic 36(2):169–200, 2000.
studies and metabolic characterization of isolates, J Wildl Dis 55. Mehlitz D: Investigations on naturally occurring Trypanosoma
19(4):308–314, 1983. evansi infections in horses, cattle, dogs and capybaras (Hydrochoe-
48. Cueto GR: Diseases of capybara. In Capybara, New York, 2013, rus hydrochaeris) in Pantanal de Poconó (Mato Grosso, Brazil),
Springer, pp 169–184. Acta Trop 58:159–169, 1994.
49. Pezzone N, Eberhardt AT, Garbaccio S, et al: Mycobacterium 56. Eberhardt AT, Monje LD, Zurvera DA, et al: Detection of
intracellulare infection in a capybara (Hydrochoerus hydrochaeris), Trypanosoma evansi infection in wild capybaras from Argentina
J Zoo Wildl Med 44(4):1098–1101, 2013. using smear microscopy and real-time PCR assays, Vet Parasitol
50. Mol JPS, Carvalho TF, Fonseca AA, et al: Tuberculosis Caused by 202(3):226–233, 2014.
Mycobacterium bovis in a Capybara (Hydrochoerus hydrochaeris), 57. Colombo VC, Nava S, Antoniazzi LR, et al: Veterinary parasitol-
J Comp Pathol 155(2):254–258, 2016. ogy factors affecting patterns of Amblyomma triste (Acari: Ixodi-
51. Alvarez JD, Moriena RA, Ortiz MI, et al: Hallazgo de Fasciola dae) parasitism in a rodent host, Vet Parasitol 211(3–4):251–258,
hepatica (Trematoda: Digenea) en un carpincho (Hydrochoerus 2015.
hydrochaeris) de la Provincia de Corrientes, Argentina, Rev Vet 58. Vasconcelos M, Martins R, Maria S: Natural infection with zoo-
20:132–134, 2009. notic subtype of Cryptosporidium parvum in capybara (Hydro-
52. Bellato V, De Souza AP, Sartor AA, et al: Ocorrência de Fasciola choerus hydrochaeris) from Brazil, Vet Parasitol 147:166–170,
hepatica na população de capivaras (Hydrochoerus hydrochaeris) 2007.
75
Xenarthra Immobilization
and Restraint
GIANMARCO ROJAS MORENO
527
528 SE C T I O N 14 Small Mammals
TABLE
75.1 Anesthetic Protocols Used in Xenarthrans
Continued
530 SE C T I O N 14 Small Mammals
TABLE
75.1 Anesthetic Protocols Used in Xenarthrans—cont’d
TABLE
75.1 Anesthetic Protocols Used in Xenarthrans—cont’d
administered, the animal should be handled as little as a Doppler vascular flow sensor over the radial artery, with a
possible and preferably left inside a containment box or 5-cm pediatric blood pressure cuff placed on the arm of the
transport box until the maximum induction effect of the animal and connected to a sphygmomanometer.17 Sloths
anesthetics has been achieved. When the anesthetics take present extreme variation in blood pressure, and stress may
effect, the sloth may be held with gloves to avoid bites. have great influence on these values.23 Pulse oximetry is
Although a sedated sloth is unable to grasp or hold on to used in sloths for monitoring cardiorespiratory function;
a branch, it is very possible that it may still bite (although however, their values may be influenced by hypothermia,
with less force). This is especially true when manipulating hypotension, and tissue pigmentation.2 The positions of the
sloths of the genus Choloepus. Sloths may also be held pulse oximeter sensor in sloths include the tongue, jaw,24
and sedated by using masks and an inhalation anesthetic; preputial area, and rectal mucosa.17
however, sloths may hold their breath for prolonged periods
of time, making this a very difficult option.2,22 A variety Anesthetic Recovery in Sloth
of injectable anesthetic combinations have been used in The use of alpha-2 antagonists and flumazenil is recom-
sloths (see Table 75.1).16,17,23–26 The reports of the use of mended in sloths, both to minimize the negative effects
anesthetic combinations include sodium pentobarbital used on the cardiovascular system and to initiate the anesthetic
alone or in association with acepromazine.27,28 Currently recovery of the animal.2,24 Sloths may require confine-
this combination is not used due to its notable depressant ment during recovery until they may grab the branches
cardiorespiratory effects, prolonged recovery time, and lack on their own. Good confinement prevents the occurrence
of antagonists. Ketamine has also been used alone or in of self-inflicted injuries associated with excitement during
combination with other anesthetics such as acepromazine, recovery.
benzodiazepines, and alpha-2 agonists.16–18,24–26,29
More recently the successful use of alpha-2 antagonists Acknowledgments
such as yohimbine and atipamezole has been reported16,24,25
and benzodiazepine antagonists such as flumazenil.25 These To the staff of the Huachipa Zoological Park, especially
antagonists have greatly reduced anesthetic recovery times to Lizette Bermúdez, for the support in the access to the
in sloths. Combinations of ketamine and diazepam or collection of xenarthrans and the information offered for
midazolam are a good choice for short-term immobiliza- this chapter.
tions if the animal is not exposed to painful procedures.
The combination of ketamine with alpha-2 agonists may
be the best option for immobilization of sloths because of References
the adequate muscle relaxation, degree of analgesia, and the 1. Rojas G: Anestesia en Hormigueros (Mirmecophaga, Tamandua
possibility of reversing their effects. It has been shown that & Cyclopes). In Rojano C, Miranda L, Ávila R, editors: Manual
protocols using medetomidine and dexmedetomidine have de Rehabilitación de Hormigueros de Colombia, El Yopal, Casa-
less deleterious effects compared with combinations that nare, 2014, Fundación Cunaguaro, Geopark Colombia S.A.S,
include xylazine.16,17,24,25 In addition, other combinations Corporinoquía, pp 97–109.
such as tiletamine-zolazepam have been used with vari- 2. West G, Carter T, Shaw J: Edentates (Xenarthra). In West
able results.16 Tiletamine-zolazepam is considered safe and G, Heard DJ, Caulkett N, editors: Zoo animal and wildlife
effective for captive conditions; however, it has a prolonged immobilization and anesthesia, ed 1, Ames, IA, 2007, Blackwell
recovery.20,24 This combination promotes deep anesthesia Publishing, pp 341–346.
3. Bermúdez L: Manejo de Cyclopes didactylus en cautiverio. In
with good muscle relaxation, but in some cases it may
Rojano C, Miranda L, Ávila R, editors: Manual de Rehabilitación
lead to respiratory depression. Combination of ketamine de Hormigueros de Colombia, El Yopal, Casanare, 2014, Fun-
and alpha-2 agonists promotes dissociative anesthesia of dación Cunaguaro, Geopark Colombia S.A.S, Corporinoquía,
approximately 40-minute duration.6,17 Combinations of pp 86–91.
ketamine, alpha-2 agonists, and benzodiazepines provide 4. Divers BJ: Edentata. In Fowler ME, editor: Zoo and wild animal
more durable anesthesia times reaching greater than 60 medicine, ed 2, Philadelphia, PA, 1986, Saunders, pp 622–630.
minutes and very short recovery times, offering an anes- 5. Fournier-Chambrillion C, Fournier P, Vie J: Immobilization of
thetic quality superior to those of other pharmacologic wild collared anteaters with ketamine and xylazine hydrochloride,
combinations.24,25 J Wild Dis 33:795–800, 1997.
6. Miranda F, Dejuste C: Principais Enfermidades em Tamanduás
Anesthetic Monitoring in Sloths Cativos. In Miranda F, editor: Manutenção de tamanduás em
cativeiro, ed 1, São Carlos, 2012, Cubo, pp 240–255.
The HR may be monitored by auscultation with the help
7. Rojas G: Use of dexmedetomidine, midazolam, ketamine and
of a pediatric stethoscope or with the support of an electro- reversal with atipamezole for chemical immobilization of giant
cardiograph. RR is verified by counting respiratory move- anteaters (Myrmecophaga tridactyla), lesser anteaters (Tamandua
ments.17 Blood pressure may be monitored in xenarthrans tetradactyla) and silky anteaters (Cyclopes didactylus) kept in
with indirect methods because the location of intravenous captivity. In Proceedings of the 44th Annual Conference of the
and intraarterial accesses is a challenge in these animals.2 In American Association of Zoo Veterinarians, Oakland, CA, 2012,
sloths, blood pressure may be measured by the placement of p 251.
534 SE C T I O N 14 Small Mammals
8. Rojas G, Miranda F: Medicina de Tamanduaí. In Miranda F, 20. Deem SL, Fiorello CV: Capture and immobilization of free-
editor: Manutenção de tamanduás em cativeiro, ed 1, São Carlos, ranging edentates. In Heard D, editor: Zoological restraint and
2012, Cubo, pp 168–185. anesthesia, Ithaca, NY, 2002, International Veterinary Informa-
9. McNab BK: Energetics, population biology, and distribution of tion Service.
Xenarthrans, living and extinct. In Montgomery GG, editor: 21. Sinclair MD: A review of the physiological effects of alpha-2
The evolution and ecology of armadillos, sloths and vermilinguas, agonists related to the clinical use of medetomidine in small
Washington, DC, 1985, Smithsonian Institution Press, pp animal practice, Can Vet J 44(11):885–897, 2003.
219–232. 22. Irving L, Scholander PF, Grinnell SW: Experimental studies of
10. Fournier-Chambrillon C, Vogel I, Fournier P, et al: Immobiliza- the respiration of sloths, J Cell Comp Physiol 20:189–210, 1942.
tion of free-ranging nine-banded armadillos and great long-nosed 23. Gilmore DP, Da Costa CP, Duarte DPF: An update on the
armadillos with three anesthetic combinations, J Wildl Dis 36: physiology of two- and three-toed sloths, Braz J Med Biol Res
131–140, 2000. 33:129–146, 2000.
11. Hernandez SM, Gammons DJ, Gottdenker N, et al: Technique, 24. Rojas G: Contención farmacológica de perezosos de dos dedos
safety, and efficacy of intra-abdominal transmitters in nine- Choloepus hoffmanni (Peters, 1858) mediante el uso de ketamina,
banded armadillos, J Wildl Manage 74(1):174–180, 2010. dexmedetomidina y midazolam, y reversión con atipamezol,
12. Orozco MM: Inmovilización química de armadillos de tres Edentata 12:20–27, 2011.
bandas (Tolypeutes matacus) mediante el uso de dos protocolos 25. Lescano J, Quevedo M, Baselly L, et al: Inmovilización química
anestésicos en el Norte Argentino, Edentata 12:1–6, 2011. reversible de corta duración en perezosos de dos dedos (Choloepus
13. Rojas G, Bermúdez L, Enciso M: Inmovilización Química de didactylus) cautivos empleando ketamina, xilacina y midazolam,
Armadillos Peludos Andinos Chaetophractus nationi (Thomas, Rev Inv Vet Perú 25(2):171–181, 2014.
1894): Uso de Ketamina, Xilacina y Midazolam con Reversión 26. Kinney ME, Cole GA, Vaughan C, et al: Physiologic and
con Yohimbina, Edentata 14:51–57, 2013. serum biochemistry values in free-ranging Hoffmann’s two-toed
14. Falzone M, Zalazar R, Gachen G, et al: Inmovilización química (Choloepus hoffmanni) and brown-throated three-toed (Bradypus
de tres tatú carreta (Priodontes maximus) en cautiverio, Edentata variegatus) sloths immobilized using dexmedetomidine and
14:66–69, 2013. ketamine, J Zoo Wildl Med 44(3):570–580, 2013.
15. Miranda FR, Costa AM: Xenarthras (tamanduás, tatu e pre- 27. Toole JF: Blood chemistry of the sloth (Choloepus hoffmanni and
guiça). In Cubas ZS, Silva JCR, Dias JL, editors: Tratado de Bradypus tridactylus), Lab Anim Sci 22:118–121, 1972.
animais selvagens: medicina veterinária, ed 1, São Paulo, 2007, 28. Meritt DA, Jr: A further note on the immobilization of sloths
Roca, pp 402–414. Choloepus spp, Int Zoo Yearb 14:160–161, 1974.
16. Vogel I, de Thoisy B, Vié JC: Comparison of injectable anesthetic 29. Rappaport AB, Hochman H: Cystic calculi as a cause of recurrent
combinations in free-ranging two-toed sloths in French Guiana, rectal prolapse in a sloth (Choloepus sp.), J Zoo Wildl Med 19:
J Wildl Dis 34:555–566, 1998. 235–236, 1988.
17. Hanley C, Siudak-Campfield J, Paul-Murphy J, et al: Immobili- 30. Sousa PC, Amorim RNL, Lima GL, et al: Establishment of an
zation of free-ranging Hoffmann’s two-toed and brown-throated anesthetic protocol for semen collection by electroejaculation in
three-toed sloths using ketamine and medetomodine: a compari- six-banded armadillos (Euphractus sexcinctus Linnaeus, 1758),
son of physiologic parameters, J Wildl Dis 44:938–945, 2008. Arq Bras Med Vet Zootec 68(6):1595–1601, 2016.
18. Gillespie DS: Xenarthra: edentata (anteaters, armadillos, sloths). 31. Aguilar RF, Superina M: Xenarthra. In Miller RE, editor: Fowler’s
In Fowler ME, Miller RE, editors: Zoo and wild animal medicine, Zoo and wild animal medicine, ed 8, St. Louis, MO, 2015,
ed 5, Philadelphia, PA, 2003, Saunders, pp 397–407. Elsevier, pp 355–368.
19. Tranquilli WJ, Thurmon JC, Grimm KA: Lumb & Jones’ veteri-
nary anesthesia and analgesia, ed 4, Ames, IA, 2007, Blackwell
Publishing.
SECTION 15
Carnivores
76 Update on Field Anesthesia Protocols for
Free-Ranging African Lions, 536
535
76
Update on Field Anesthesia Protocols
for Free-Ranging African Lions
PETER BUSS AND MICHELE MILLER
536
CHAPTER 76 Update on Field Anesthesia Protocols for Free-Ranging African Lions 537
TABLE Immobilizing Drugs, Drug Combinations, and Antagonists Commonly Used in Free-Ranging African
76.1 Lions (Doses Are Given in mg/kg or as a Ratio [Antagonist:Agonist] in mg)
Detomidine (IM)
Midazolam (IM)
Ketamine (IM)
3–52
4–5 (≤10)4
0.38–1.327 0.027–0.055 2.5–5× medetomidine
6
1.3–2.3 0.06–0.08 0.2–0.4
3
2.5 0.07
2
5–7 0.03–0.05 5× medetomidine
3
4–5 0.05
8
0.3 0.05 0.2 0.7 0.3 0.0032
2
0.2–0.3 0.05 0.15 2× butorphanol 5× medetomidine
is vulnerable to attack by other lions and hyenas, and may administration, and whether additional doses were given.
suffer injuries as it persistently tries to stand before it is ready. Recovery times are reduced if the antidote is given later in
the immobilization period as a result of tiletamine metabo-
Cyclohexylamine Plus Alpha2-Agonist lism. Lions immobilized with lower TZM doses recover to
walking within 8–30 minutes following atipamezole admin-
The dose of TZ required to immobilize lions can be istration at approximately 45 minutes after induction.7
reduced by as much as 75% if combined with medetomi- The use of ketamine plus medetomidine or detomidine
dine. Recovery times are significantly reduced by antag- in free-ranging lions has been reported.3 These drug
onizing the medetomidine effects with atipamezole (see combinations resulted in induction times of 6–10 minutes
Table 76.1).6 TZM provides a smooth anesthetic induction with limited respiratory and cardiovascular adverse effects.
within 3–10 minutes with good muscle relaxation and suf- Hemoglobin oxygen saturation varied from 85% to 90%.
ficient analgesia for minor surgical procedures.6,7 Decreases Atipamezole was administered approximately 1 hour after
in respiration and heart rate are limited and remain stable immobilization and recovery took 25–32 minutes. These
throughout the anesthesia, and hemoglobin oxygen satu- combinations are recommended for short-duration proce-
ration values seldom decrease below 90%. Rectal tempera- dures such as radio-collaring, disease surveillance, or snare
ture may exceed 40°C and should be closely monitored. The removal.3
palpebral reflex may be present and becomes more promi-
nent as the duration of anesthesia increases.7 Spontaneous Butorphanol, Medetomidine, and Midazolam
recoveries may occur after approximately 1 hour following
induction, and repeat intramuscular (IM) doses of both BMM combination (see Table 76.1) provides an effective
TZ and medetomidine (one-third of induction dose) are immobilizing drug combination for lions with a rapid induc-
recommended. In the authors’ experience, multiple addi- tion (5–10 minutes) and 45 minutes of stable anesthesia.8
tional TZM doses may result in prolonged recovery times Immobilized lions do not react to minor painful procedures
as a consequence of accumulative tiletamine effect and judi- but more invasive surgeries require additional analgesia.
cious administration is advised. Mild bradycardia (<50 beats/min), mild metabolic acidosis,
Recovery to standing and walking is variable depending normocapnia, and mean PaO2 of 87 mm Hg have been
on initial TZM dose, time between darting and atipamezole reported in BMM-immobilized lions.8 Complete reversal
538 SE C T I O N 15 Carnivores
is achieved with naltrexone-atipamezole-flumazenil admin- tonicity. Alighting from a helicopter and approaching an
istration, with recovery to standing within 4–8 minutes. An apparently immobilized lion should be done with extreme
advantage of this combination is that reversal agents can be caution until the anesthetic depth may be confirmed.
administered at any time point following induction.8
Field experience has shown that BMM-immobilized
lions may spontaneously stand up at ≥45 minutes after References
induction, especially following a loud noise or painful
1. Quandt SKF: Immobilization of African lions Panthera leo
manipulation. Immobilized lions should be administered with medetomidine/ketamine, in comparison with tiletamine/
one-third of the initial BMM dose IM at 45 minutes after zolazepam and phencyclidine. In Proceedings of Capture, Care
induction and every subsequent 30 minutes. The use of & Management of Threatened Mammals, Skukuza, Kruger
blindfolds and hobbles is advocated to limit the possibility National Park, 1993, pp 64–75.
of injury to people working with the lions. Antidote dose 2. Burroughs R, Hofmeyr M, Morkel P, et al: Chemical immobiliza-
rates are calculated on the total immobilizing drug doses tion – Individual species requirements. In Kock MD, Burroughs
administered. The authors have not observed adverse central R, editors: Chemical and physical restraint of wild animals: a
nervous system or cardiorespiratory effects following intra- training and field manual for African species, ed 2, 2012, IWVS
venous atipamezole administration in cases of anesthetic (Africa), pp 143–264.
emergencies. Additional atipamezole doses (10× medeto- 3. Fyumagwa RD, Bugwesa ZK, Mdaki ML, et al: Comparison of
anaesthesia and cost of two immobilization protocols in free-
midine dose, mg) may also be given following prolonged
ranging lions, S Afr J Wildl Res 42:67–70, 2012.
procedures to ensure animals do not become resedated. 4. McKenzie AA: Chemical capture of carnivores. In McKenzie AA,
At the suggested midazolam dose rates, the omission of editor: The capture and care manual: capture, care, accommodation
flumazenil does not significantly change the quality or time and transportation of wild African animals, 1993, Wildlife Deci-
to recovery. sion Support Services, The South African Veterinary Foundation,
pp 224–253.
5. Stegman GF, Bester L, Venter L: Halothane anaesthesia in an
Darting From a Helicopter African lion, S Afr J Wildl Res 30:93–95, 2000.
Helicopter-based darting of lions should only be attempted 6. Jacquier M, Aarhaug P, Arnemo JM, et al: Reversible immo-
by a capable pilot and experienced veterinarian. This bilization of free-ranging African lions (Panthera leo) with
capture method results in a marked sympathetic response medetomidine-tiletamine-zolazepam and atipamezole, J Wildl
Dis 42:432–436, 2006.
in the lion, and the authors have found TZM and BMM
7. Fahlman Å, Loveridge A, Wenham C, et al: Reversible anaesthe-
to be ineffective unless administered at more than twice sia of free-ranging lions (Panthera leo) in Zimbabwe, J S Afr Vet
the recommended dose. TZ is the suggested drug of Assoc 76:187–192, 2005.
choice; however, hyperthermia in the immobilized lion is 8. Wenger S, Buss P, Joubert J, et al: Evaluation of butorphanol,
a significant risk as a result of increased exertion caused by medetomidine and midazolam as a reversible narcotic combina-
the approach of the helicopter during darting, increased tion in free-ranging African lions (Panthera leo), Vet Anaesth Analg
daytime ambient temperatures, and drug-induced muscle 37:491–500, 2010.
77
Overview of African
Wild Dog Medicine
JENNIFER N. LANGAN AND GWEN JANKOWSKI
539
540 SE C T I O N 15 Carnivores
• Figure 77.1 Distribution of the remaining African wild dog (Lycaon pictus) populations.
Permanent or even brief temporary removal of an established have access to multiple heated areas if the temperature
pack member may have profound social impacts, including regularly drops below 4.4°C–7.2°C (40°F–45°F) and should
changes in social hierarchy with aggression so substantial have shelter from the elements.7 Additionally, a heated den
that reintroduction may not be possible.7,15 Detailed plans should be provided if breeding is planned. Facilities should
to reduce stress and promote normal behaviors should be have sufficient holding space to accommodate separating
implemented if a dog must be isolated, including maintain- animals for long periods. With the exception of fish, housing
ing olfactory and visual contact. If separation is required, it African wild dogs with other species is not recommended
may be helpful to subdivide the pack and then reintroduce due to their high predatory drive.
them all simultaneously. Alternatively, introductions of sub- African wild dogs should be moved only in sturdy
ordinate dogs first, then dominant pairs, may be effective. metal or wood crates with good ventilation that meet US
Successful implementation of enrichment has included Department of Agriculture (USDA) and International Air
environmental devices, sensory stimulants, and food, Transport Association (IATA) requirements for live animal
behavioral, and habitat variance.16 Piles of leaves, dirt, and transport. Completing a written transport plan, health
mulch allow natural digging and rolling behaviors. Rotating evaluation, and crate training facilitates transitions when
exhibits with other predators provides habitat diversity and animals are relocated.
promotes scent-marking behavior. Offering carcass feeds
hung from trees or on zip lines, feeding bones, providing Nutrition
several types of enrichment to the pack simultaneously, and
permitting breeding when possible are recommended for African wild dogs are generalist predators, occupying a
this species.15,16 range of habitats where they hunt medium-sized antelope.6
Enclosures should be large and contain ample space for In natural settings, reduced prey populations and competi-
exercise to meet the animals’ physical, social, behavioral, tion from other predators inhibit population growth.3,17
and psychological needs.7 Specific size and perimeter recom- African wild dogs in zoos are fed a nutritionally complete
mendations may be found in the Association of Zoos & raw meat–based diet (1–1.36 kg/adult/day) and are
Aquariums (AZA) Large Canid Care Manual.7 Facilities that supplemented with small whole prey, knuckle/rib/shank
allow the public to observe the animals should prevent close bones (one to two times week), and carcasses (pig, deer,
contact or inadvertent access to the enclosure. Dogs should calf, horse). Lactating bitches require up to three times
CHAPTER 77 Overview of African Wild Dog Medicine 541
maintenance caloric intake. Specific recommendations for AH, Inc., Fort Worth, Texas). Current estimates show 23%
kilocalorie requirements may be found in the AZA Large of females have some degree of reproductive pathology,28
Canid Care Manual.7 Feeding African wild dogs a portion of the most frequently reported of which is cystic endometrial
their diet while separated from the pack aids in monitoring hyperplasia (CEH) with or without pyometra and adeno-
individual animals’ food consumption. Packs are generally myosis (Fig. 77.2).27–29 Adenocarcinoma, uterine rupture, and
fasted from their normal meat diet 1 day per week and may pyometra without other pathology have also been reported
be provided with bones. (Kinsel, personal communication, April 10, 2017).23,27,28 The
Species Survival Plan Program (SSP) currently recommends
Reproduction and Contraception that all postreproductive females (>10 years) be spayed.8
Deslorelin implants have been used for contraception and
African wild dogs are seasonally monoestrous obligate behavioral alteration in males with variable results (see Table
cooperative breeders with a brief copulatory tie.18,19 Within 77.1 for a summary of reproductive information) (see also
a pack the alpha male and female produce the majority of Chapter 22).26,30,31
surviving pups annually.7,20,21 Most successful reproduction
occurs after 2 years of age, with senescence around 8–9 Handling, Restraint, and Anesthesia
years.8,21 Subordinate females may reproduce, but offspring
typically do not survive. More often, subordinate females Healthy adult African wild dogs are not physically restrained
develop pseudopregnancy and may lactate in order to help due to safety concerns. Noninvasive procedures including
care for the pups of the dominant pair.1 Females produce an visual examination, hand injections, wound treatment,
average of six to eight pups22,23 and up to 21 pups8 in a den venipuncture, and crate training may be accomplished with
after a gestation of 69–71 days.24–26 Primiparous females have operant conditioning.
higher estrogen, which is reported to result in more male Restraint cages are useful and provide a safe, controlled
offspring.24 Hand-rearing is not recommended due to the environment to facilitate intramuscular injections of
extremely aggressive and social nature of these animals.8 In anesthetics. A quiet area away from the pack promotes
zoos, the breeding pair is separated from the pack to prevent quick inductions and smooth recoveries. Remote injection
trauma to the pups. The group is gradually introduced when systems are recommended for immobilizing free-ranging
the pups begin to emerge from the nest box. A birth plan African wild dogs or in situations when a chute is not avail-
detailing responses to aggression toward the pups, large litter able. Anesthetic regimens selected should take into account
size, and other contingencies is recommended. health status, age, and environmental conditions. In cases
Newborn pups weigh about 300 g, open their eyes where cardiovascular disease has been confirmed or cardiac
around 2 weeks of age, and emerge from the den to start status is unknown, alpha-2 agonists should be avoided
taking solid food at 3 weeks. Sex determination is similar to and alternatives such as ketamine-midazolam-butorphanol
that for other canids. Pups are weaned and start to follow with propofol or gas anesthesia (isoflurane, sevoflurane)
the pack at 11–12 weeks of age. In free-range settings, all should be considered. Chemical restraint protocols used
members of the pack raise the pups; they regurgitate food in African wild dogs may be found in Table 77.2 and
while the young are in the den and relinquish kills to the have been previously published.32–34 Drug combinations at
pups and yearlings.1,3 higher dosages for free-ranging African wild dogs are avail-
Reproductive anatomy is similar to that of other canid able in the literature.35,36 Reversal of alpha-2 agonists and
species. Owing to social dynamics, most reproduction has opioids with atipamezole and naloxone decreases recovery
been natural; however, semen has been preserved and used time. To avoid dysphoria, it is advised to wait at least
for artificial inseminations.15 Captive Lycaon pictus generally 60 minutes postinduction before administering reversals.
reproduce in the fall in the northern hemisphere.8 Estrus Recovery in a crate or nest box may reduce struggles to
lasts 6–9 days and includes vulvar swelling and sanguineous stand during recovery. Telazol (tiletamine HCL and zolaz-
discharge with interest from the male. Attraction from the epam HCL, Zoetis, Parsippany, New Jersey) as a sole agent
male may be observed for 1–2 weeks prior to tying.14,25 or used in combination with medetomidine is a reliable
Males show increased testicular development, spermator- option during an emergency response but often results in
rhea, and semen production; therefore the corresponding a prolonged recovery time.34 Vascular access, intubation,
seasonal ability to collect sperm via electroejaculation is and monitoring equipment are applied as in other canids.
improved.15 Every wild dog anesthesia should include oxygen supple-
The progestin-based melengestrol acetate (MGA) implant, mentation, electrocardiography (ECG), and monitoring of
previously used in canids, has been associated with uterine pulse oximetry, heart and respiratory rate, blood pressure,
pathology.27 The AZA Reproduction Management Center (for- and temperature.
merly the Wildlife Contraception Center) (www.stlzoo.org/
animals/scienceresearch/reproductivemanagementcenter) Clinical Pathology
recommends gonadotropin releasing hormone (GnRH)
agonists such as Suprelorin (deslorelin acetate) implants or As in other canids, the jugular, cephalic, and saphenous
Lupron Depot (leuprolide acetate 4.7 mg implant, Virbac veins are commonly used sites for venipuncture. Tables 77.3
542 SE C T I O N 15 Carnivores
A B
C D
• Figure 77.2 Common uterine pathology in African wild dogs (Lycaon pictus): (A) uterine hyperplasia
and pyometra, (B) uterine adenocarcinoma with associated pyometra, (C) ultrasound image of a dilated
uterine horn filled with hyperechoic material (arrow), and (D) and corresponding intraoperative image in
the same animal with pyometra.
TABLE Reproductive Information for African Wild extinction events.38–41 There is serologic evidence of expo-
77.1 Dog (Lycaon pictus) sure to canine parvovirus, canine distemper virus, adeno-
virus, rabies virus, coronavirus, rotavirus, and Ehrlichia
Reproductive Cycle Monestrous canis.21,42,43 Outbreaks of anthrax in Kruger National Park
Usual age of first reproduction 21–22 months occur, but infected animals commonly survive.43 Contact
with domestic dogs has been reported to increase exposure
Copulation (North America) August–October
to some canid pathogens, but sylvatic viral strains also pose
Length of estrus 6–9 days a significant threat.5,44
Gestation (from first day of 69–71 days Parasitic disease and infection has rarely been described.44–46
copulation) Toxocara canis, Dipylidium caninum, Spirometra sp., Taeni-
Parturition (North America) October–January
idae, and Ancylostoma spp., as well as two genera of canid
protozoan gastrointestinal parasites, Sarcocystis and Isopora,
Mean/maximum litter size 6–8/21 were identified in fecal samples from free-ranging animals
but were not associated with clinical disease.44,47 Standard
anthelmintics at canine dosages have been successfully used
and 77.4 show normal hematologic and serum chemistry to treat internal parasites. It is recommended that African
reference ranges for captive African wild dogs.9,37 wild dogs be routinely tested and maintained on prophy-
lactic heartworm preventative in endemic areas.
Diseases Over the last decade, valvular dysplasia of varying sever-
ity has been increasingly recognized as a significant concern
Rabies and distemper have contributed to mortality in in African wild dogs in North America. Sibling groups
African dog populations, occasionally resulting in local and offspring have been affected over multiple generations,
CHAPTER 77 Overview of African Wild Dog Medicine 543
TABLE
77.2 Select Chemical Restraint Agents Used in African Wild Dogs (Lycaon pictus)
TABLE Hematologic Values for African Wild Dogs echocardiographic examination with preventative health
77.3 (Lycaon pictus) evaluations for captive wild dogs is recommended by the
SSP in order to identify and medically manage affected
Parameter Mean SD individuals.8 Treatment recommendations are based on
White blood cell count 10.7 3.53 those for domestic dogs and have included angiotensin-
(×103/µL) converting enzyme (ACE) inhibitors, diuretics, and inodila-
Red blood cell count 7.98 1.61 tors (pimobendan). Despite the challenges associated with a
(×106/µL) progressive genetic bottleneck, it is strongly recommended
Hemoglobin (g/dL) 15.1 2.43
that individuals with cardiac disease not be selected for
breeding to prevent further incorporation of genetic defects
Hematocrit (%) 43.7 7.33 into breeding lines (Briggs, personal communication, April
MCV (fL) 55.6 4.53 22, 2017).
MCH (pg/cell) 19 1.75
Neoplasia has not been extensively reported in the
literature but appears to play an important role in the
MCHC (g/dL) 34.1 2.08 health of captive populations.48 Apocrine gland tumors have
Platelet count (×103/µL) 451 183.8 been documented in clinical settings, presenting as single
Segmented neutrophils 7.44 3.34
or multiple dorsal cutaneous masses that can progress to
(×103/µL) large regionally invasive tumors (Fig. 77.4) (Agnew, per-
sonal communication, April 25, 2017). Cases from females
Neutrophilic bands (×103/µL) 0.508 0.025
are overrepresented in pathology reports submitted to the
Lymphocytes (×10 /µL) 3
1.9 5.37 African wild dog SSP (Kinsel, personal communication,
Eosinophils (×10 /µL) 3
0.458 0.396 April 10, 2017). Surgical excision is recommended, although
3
large tumors may require other forms of treatment. Tumor
Basophils (×10 /µL) 0.014 0.039
growth results in ulceration and necrosis and negatively
General ZIMS database reference: Species360 (2017), ZIMS.Species360 affects an animal’s quality of life. Other common neoplasias
.org. include hemangiosarcoma, peripheral odontogenic fibroma
(fibromatous epulis), adrenocortical adenoma/carcinoma,
and mammary and uterine neoplasia.
suggesting that the condition has a genetic, inheritable Other notable conditions diagnosed in African wild
basis, as is well documented in domestic dogs.8 Cases range dogs include dental disease—particularly fractured teeth—
from minor to severe valvular insufficiency with congestive pancreatitis, diabetes, spina bifida, syringomyelia, keratitis,
heart failure (Fig. 77.3). Mildly affected animals exhibit no snake bites,49 and trauma from conspecifics (Kinsel, personal
clinical signs, making the condition difficult to detect. The communication, April 10, 2017). African wild dogs mask
extent of this disease is uncertain, but it is highly prob- signs of illness and may have advanced disease by the time
able that cardiac disease is underdiagnosed. Incorporating a change in behavior or appetite is observed.
544 SE C T I O N 15 Carnivores
Parameter Mean SD
Calcium (mg/dL) 10.2 0.78
Phosphorus (mg/dL) 5.5 1.88
Sodium (mEq/L) 148 4.75
Potassium (mEq/L) 4.6 0.75
Chloride (mEq/L) 116 5.25
A
Bicarbonate (mEq/L) 20 5.25
Carbon dioxide (mEq/L) 19.6 4.25
Blood urea nitrogen (mg/dL) 25 8.75
Creatinine (mg/dL) 1.1 0.35
Total bilirubin (mg/dL) 0.2 0.1
Glucose (mg/dL) 148 39.3
Cholesterol (mg/dL) 260 74
Triglyceride (mg/dL) 69 43.25
Creatine phosphokinase (IU/L) 229 141.75
Alkaline phosphatase (IU/L) 55 45.25
Alanine aminotransferase (IU/L) 50 21
Aspartate aminotransferase 36 15.75
(IU/L)
Gamma glutamyltransferase 6 3
(IU/L)
Lactate dehydrogenase (IU/L) 181 172
Uric acid (mg/dL) 0.4 0.35 B
Amylase (IU/L) 375 199.5
Lipase (IU/L) 123 70.5
Total protein (colorimetry) (g/dL) 5.9 0.63
Globulin (colorimetry) (g/dL) 2.8 0.6
Albumin (colorimetry) (g/dL) 3.2 0.4
Preventative Medicine
Preventative health, preshipment, and quarantine examina-
tions should be conducted to determine an animal’s health.
A complete physical exam—including a dental examination,
complete blood count, chemistry panel, heartworm testing,
urinalysis, radiographs of the thorax and abdomen, and
evaluation for endo- and ectoparasites—should be com-
pleted on a routine schedule as part of a preventive medicine
plan. Additionally, abdominal ultrasound examination or D
computed tomography in females and echocardiograms for • Figure 77.3 Images from African wild dogs (Lycaon pictus) with
both sexes are recommended owing to disease predilection heart disease: (A) Lateral and (B) ventrodorsal radiographs of the
in this species. thorax consistent with congestive heart failure. (C) Echocardiography
image showing mitral insufficiency. (D) Necropsy image of mitral valve
dysplasia.
CHAPTER 77 Overview of African Wild Dog Medicine 545
External and internal parasites should be treated accord- Athens, Georgia) provided persistent protective titers.57
ing to domestic dog guidelines. Fleas, ticks, and ear tip A survey in 2006 showed that most African wild dogs
trauma from biting flies have responded well to products maintained presumably protective titers after vaccination
containing carbaryl or pyrethrins. Good hygiene, removing for canine distemper and rabies for 1 year; however, few
standing water, fly traps, and premise sprays help control dogs maintained titers for 2–3 years.58 There is a paucity of
fly and mosquito populations. scientific information regarding vaccination against canine
Newly acquired animals should be quarantined away parvovirus and leptospiral infection. Protocols should be
from the collection for a predetermined period based on developed that take into consideration local environmental
a thorough risk assessment by the supervising veterinarian disease prevalence, animal health, and risk factors. Select
and have at least two negative fecal examinations. Animals vaccination protocols for captive African wild dogs are listed
should be individually identified with microchip transpon- in Table 77.5 (see also Chapter 79).
ders placed subcutaneously between the shoulder blades or
to the left of midline over the shoulder. Acknowledgments
Currently there are no universal recommendations for
vaccination protocols. The safety and efficacy of vaccines This chapter is dedicated to the researchers, veterinarians,
in African wild dogs have historically been unsatisfactory. biologists, and animal care staff that have contributed to
Vaccination strategies to conserve free-ranging populations our collective knowledge, helped conserve wild populations,
have been reported and continue to be investigated.50 and have improved the care of African wild dogs around the
Modified live canine distemper vaccinations have failed world. We thank Drs. Anneke Moresco and Mike Kinsel for
to produce protective antibody levels in some cases51 and their contributions to reproduction, pathology, and disease
have induced distemper resulting in mortality in other presented in this chapter and Drs. Sathya Chinnadurai and
cases.7,45,52,53 Vaccine-induced distemper can be avoided by Matthew Lenyo for sharing their expertise and thoughtful
using killed vaccines, and at least one vaccine (Purevax review.
ferret Distemper, Merial Inc., Athens, Georgia) has been
shown to produce measurable titers after a series of three References
injections at 2- to 3-week intervals.54 Subunit canine dis-
temper vaccines (CDV-ISCOM, Erasmus MC, Rotterdam, 1. Creel S, Creel NM: The African wild dog: behavior, ecology and
The Netherlands) stimulated appropriate titer formation, conservation, Princeton, NJ, 2002, Princeton University Press.
but titers did not endure relative to the Purevax Ferret 2. Sillero-Zubiri C, Hoffmann M, Macdonald DW: Canids: foxes,
Distemper vaccine. Vaccine recommendations for domestic wolves, jackals and dogs. Status survey and conservation action
dogs have been reduced from yearly to triennially; however, plan. IUCN/SSC Canid Specialist Group, Gland, Switzerland
it is unknown whether nondomestic canids maintain titers and Cambridge, UK, 2004, IUCN.
3. Hutchins M, Evans A, Jackson J, et al: Carnivora. In Grzimek’s
in a similar manner. In one study, protective titers from
animal life encyclopedia, ed 2, 17 vols. Detroit, MI, 2003, Gale
the Purevax vaccine persisted in 39%–85% of African Virtual Reference Library, pp 265–278.
wild dogs for a minimum of 1 year.55 Early studies have 4. Woodroffe R, Sillero-Zubiri C 2012. Lycaon pictus (North Africa
indicated that vaccination with killed rabies vaccines may subpopulation). The IUCN Red List of Threatened Species 2012:
not be sufficient for protection.5,50,56 However, a recent e.T16991111A16991120. http://dx.doi.org/10.2305/IUCN.UK
study showed that a single intramuscular vaccination .2012.RLTS.T16991111A16991120.en. (Accessed 10 April
of dogs older than 14 weeks with Imrab 3 (Merial Inc., 2017).
546 SE C T I O N 15 Carnivores
5. Woodroffe R, Prager KC, Munson L, et al: Contact with domes- 24. Creel S, Creel NM, Monfort SL: Birth order, estrogens and
tic dogs increases pathogen exposure in endangered African wild sex-ratio adaptation in African wild dogs (Lycaon pictus), Anim
dogs (Lycaon pictus), PLoS ONE 7(1):e30099, 2012, doi:10.1371/ Reprod Sci 53:315–320, 1998.
journal.pone.0030099. 25. Monfort SL, Wasser SK, Mashburn KL, et al: Steroid metabo-
6. Moreangels MM, Marino J, Groom RJ: Diet of four sympatric lism and validation of noninvasive endocrine monitoring in
carnivores in Save Valley Conservancy, Zimbabwe: implications the African wild dog (Lycaon pictus), Zoo Biol 16:533–548,
for conservation of the African wild dog (Lycaon pictus), S Afr J 1997.
Wildl Res 42:94–103, 2012. 26. Newell-Fugate AE, Nöthling JO, Bertschinger J: Seasonal changes
7. AZA Canid TAG: Large canid (canidae) care manual, Silver in steroid hormone profiles, body weight, semen quality and the
Spring, MD, 2012, Association of Zoos & Aquariums. reproductive tract in captive African wild dogs (Lycaon pictus) in
8. Quick M, Rhodes S, Sullivan S: Population analysis and breeding South Africa, Gen Comp Endocrinol 178:272–281, 2012.
and transfer plan: african painted (wild) dog (Lycaon pictus). 27. Moresco A, Munson L, Gardner IA: Naturally occurring and
AZA Species Survival Plan Yellow Program. [www.aza.org], melengestrol acetate-associated reproductive tract lesions in zoo
2017. canids, Vet Path 46:1117–1128, 2009.
9. General ZIMS database reference: Species360, ZIMS.Species360. 28. Lamglait B, Trunet E, Leclerc A: Retrospective study of mortality
org, 2017. of captive African wild dogs (Lycaon pictus) in a French zoo
10. Potgieter KR, O’Riain MJ, Davies-Mostert HT: Behavioural cues (1974–2013), J Zoo Aquar Res 3:47–51, 2015.
can be used to predict the outcome of artificial pack formation 29. Jankowski G, Adkesson MJ, Langan JN, et al: Cystic endometrial
in African wild dogs (Lycaon pictus), Afr J Wildl Res 45:215–222, hyperplasia and pyometra in three captive African hunting dogs
2015. (Lycaon pictus), J Zoo Wildl Med 43:95–100, 2012.
11. Van der Weyde LK, Martin GB, Paris MC: Monitoring stress 30. Bertschinger HJ, Asa CE, Calle PP, et al: Control of reproduction
in captive and free-ranging African wild dogs (Lycaon pictus) and sex related behaviour in exotic wild carnivores with the
using faecal glucocorticoid metabolites, Gen Comp Endocrinol GnRH analogue deslorelin: preliminary observations, J Reprod
226:50–55, 2016. Fertil Suppl 57:275–283, 2001.
12. Monfort SL, Mashburn KL, Brewer BA, et al: Evaluating adrenal 31. Bertschinger HJ, Trigg TE, Jöchle W, et al: Induction of con-
activity in African wild dogs (Lycaon pictus) by fecal corticosteroid traception in some African wild carnivores by downregulation
analysis, J Zoo Wildl Med 29:129–133, 1998. of LH and FSH secretion using the GnRH analogue deslorelin,
13. Creel S, Creel NM, Monfort SL: Social stress and dominance, Reprod Suppl 60:41–52, 2002.
Nature 379:212, 1996. 32. Van Heerden J, Burroughs RE, Dauth J, et al: Immobilization
14. Van der Weyde LK, Martin GB, Blackberry MA, et al: Repro- of wild dogs (Lycaon pictus) with a tiletamine hydrochloride/
ductive hormonal patterns in pregnant, pseudopregnant and zolazepam hydrochloride combination and subsequent evaluation
acyclic captive African wild dogs (Lycaon pictus), Anim Reprod of selected blood chemistry parameters, J Wildl Dis 27:225–229,
Sci 156:75–82, 2015. 1991.
15. Johnston SD, Ward D, Lemon J, et al: Studies of male reproduc- 33. Ward DG, Blyde D, Lemon J, et al: Anesthesia of captive African
tion in captive African wild dogs (Lycaon pictus), Anim Reprod Sci wild dogs (Lycaon pictus) using a medetomidine-ketamine-
100:338–355, 2007. atropine combination, J Zoo Wildl Med 37:160–164, 2006.
16. Cloutier TL, Packard JM: Enrichment options for African 34. Larsen RS, Kreeger TJ: Canids. In West G, Heard D, Caulkett
painted dogs (Lycaon pictus), Zoo Biol 33:475–480, 2014. N, editors: Zoo animal and wildlife immobilization and anesthesia,
17. Nowak RM: Order carnivora. In Walker’s mammals of the world, ed 2, Ames, IA, 2014, Wiley-Blackwell, pp 585–598.
(vol 1), ed 6, Baltimore, MD, 1999, Johns Hopkins University 35. Kreeger TJ: Chemical restraint and immobilization of wild
Press, pp 676–678. canids. In Fowler ME, Miller RE, editors: Zoo & wild animal
18. Asa C: Hormonal and experiential factors in the expression of medicine, current therapy, ed 4, Philadelphia, PA, 1999, Saunders,
social and parenteral behavior of canids. In Soloman N, French pp 429–435.
J, editors: Cooperative breeding in mammals, Cambridge, 1996, 36. Osofsky SA, McNutt JW, Hirsch KJ: Immobilization of free-
Cambridge University Press. ranging African wild dogs (Lycaon pictus) using a ketamine/
19. Malcolm JR, Marten K: Natural selection of the communal xylazine/atropine combination, J Zoo Wildl Med 27:528–532,
rearing of pups in African wild dogs (Lycaon pictus), Behav Ecol 1996.
Sociobiol 10:1–13, 1982. 37. Teare JA, editor: 2103, Lycaon pictus, no selection by gender,
20. Girman D, Mills MGL, Geffen E, et al: A molecular genetic all ages combined, 2013 CD.html in ISIS Physiological Ref-
analysis of social structure, dispersal, and interpack relations erence Intervals for Captive Wildlife: A CD-ROM Resource,
of the African wild dog (Lycaon pictus), Behav Ecol Sociobiol International Species Information System, Eagan, MN.
40:187–198, 1997. 38. Alexander KA, Appel MJ: African wild dogs (Lycaon pictus)
21. Creel S, Creel NM, Munson L, et al: Serosurvey for selected endangered by a canine distemper epizootic among domestic
viral diseases and demography of African wild dogs in Tanzania, dogs near the Masai Mara National Reserve, Kenya, J Wildl Dis
J Wildl Dis 33:823–832, 1997. 30:481–485, 1994.
22. Frantzen MAJ, Ferguson JWH, De Villiers MS: Conservation 39. Alexander KA, Kat PW, Munson LA, et al: Canine distemper-
role of captive African wild dogs (Lycaon pictus), Biol Conserv related mortality among wild dogs (Lycaon pictus) in Chobe
100:253–260, 2001. National Park, Botswana, J Zoo Wildl Med 27:426–427, 1996.
23. Newell-Fugate A, Lane E: Intrapartum uterine rupture with 40. Goller KV, Fyumagwa RD, Nikolin V, et al: Fatal canine dis-
coincidental uterine adenomyosis in an African wild dog (Lycaon temper infection in a pack of African wild dogs in the Serengeti
pictus), J Zoo Wildl Med 40:791–795, 2009. ecosystem, Tanzania, Vet Microbiol 146:245–252, 2010.
CHAPTER 77 Overview of African Wild Dog Medicine 547
41. Gascoyne SC, Laurenson MK, Lelo S, et al: Rabies in African 50. Prager KC, Woodroffe R, Cameron A: Vaccination strategies to
wild dogs (Lycaon pictus) in the Serengeti region, Tanzania, J conserve the endangered African wild dog (Lycaon pictus), Biol
Wildl Dis 29:396–402, 1993. Conserv 144:1940–1948, 2011.
42. Alexander KA, Conrad PA, Gardner IA, et al: Serologic survey 51. Van Heerden J, Swart WH, Meltzer DGA: Serum antibody levels
for selected microbial pathogens in African wild dogs (Lycaon before and after administration of live canine distemper vaccine
pictus) and sympatric domestic dogs (Canis familiaris) in Maasai to the wild dog (Lycaon pictus), J S Afr Vet Med Assoc 51:283–284,
Mara, Kenya, J Zoo Wildl Med 24:140–144, 1993. 1980.
43. Van Heerden J, Mills MG, van Vuuren MJ, et al: An investiga- 52. McCormick AE: Canine distemper in African hunting dogs
tion into the health status and diseases of wild dogs (Lycaon (Lycaon pictus) – possibly vaccine induced, J Zoo Anim Med
pictus) in Kruger National Park, J S Afr Vet Assoc 66:18–27, 1995. 14:66–71, 1983.
44. Flacke G, Spiering P, Cooper D, et al: A survey of internal 53. Durchfeld B, Baumgartner W, Herbst W, et al: Vaccine-associated
parasites in free-ranging African wild dogs (Lycaon pictus) from canine distemper infection in a litter of African hunting dogs
KwaZulu-Natal, South Africa, S Afr J Wildl Res 40:176–180, (Lycaon pictus), Zentralblatt für Veterinrmedizin 37:203–212,
2010. 1990.
45. Van Heerden J, Bainbridge N, Burroughs RE, et al: Distemper- 54. Connolly M, Thomas P, Woodroffe R, et al: Comparison of oral
like disease and encephalitozoonosis in wild dogs (Lycaon pictus), and intramuscular recombinant canine distemper vaccination in
J Wildl Dis 25:70–75, 1989. African wild dogs (Lycaon pictus), J Zoo Wildl Med 44:882–888,
46. Matjila PT, Leisewitz AL, Jongejan F, et al: Molecular detection 2013.
of Babesia rossi and Hepatozoon sp. in African wild dogs (Lycaon 55. Philippa J, Visee A, van de Bildt M, et al: Humoral immune
pictus) in South Africa, Vet Parasitol 157:123–127, 2008. response of African wild dogs (Lycaon pictus) to novel canine
47. Berentsen AR, Becker MS, Stockdale-Walden H, et al: Survey of distemper vaccines in Proceedings of the American Association
gastrointestinal parasite infection in African lion (Panthera leo), of Zoo Veterinarians, 254, 2006.
African wild dog (Lycaon pictus) and spotted hyaena (Crocuta 56. Scheepers JL, Venzke KAE: Attempts to reintroduce African wild
crocuta) in the Luangwa Valley, Zambia, Afr Zool 47:363–368, dogs (Lycaon pictus) into Etosha National Park, Namibia, S Afr J
2012. Wildl Res 25:138–140, 1995.
48. McAloose D, Raske M, Moore R, et al: Multilobular tumor of 57. Connolly M, Thomas P, Woodroffe R, et al: Single versus double
bone in an African wild dog (Lycaon pictus), J Zoo Wildl Med dose rabies vaccination in captive African wild dogs (Lycaon
43:950–952, 2012. pictus), J Zoo Wildl Med 46:691–698, 2015.
49. Weise FJ, van Vuuren RJ, Echement KE, et al: Successful snake- 58. Zollinger T, Gamble K, Barbiers R: Immunization and antibody
bite treatment in three juvenile African wild dogs (Lycaon pictus) persistence to canine distemper and rabies vaccination in captive
with polyvalent antivenom: A Namibian case report, J S Afr Vet african wild dogs (Lycaon pictus), in Proceedings of the American
Assoc 84(1):2013, Art. #982, 4 pages. http://dx.doi.org/10.4102/ Association of Zoo Veterinarians, 393, 2006.
jsava.v84i1.982.
78
Medicine of Captive Andean Bears
LEONARDO ARIAS-BERNAL AND ENRIQUE YARTO-JARAMILLO
548
CHAPTER 78 Medicine of Captive Andean Bears 549
In one study, two Andean bears fed a mixed diet of been performed (A Moresco, K Herrick, personal com-
produce and commercial dry feed showed dry matter munication, February 28, 2017). Fecal monitoring of sex
digestibility coefficients of 60.5%. In the same study, the steroids, specifically estradiol and progesterone, has been
bears consumed 1.6% of body weight in dry matter daily. carried out in six captive spectacled bear females in Peru.22
Apparent digestibility coefficients for neutral detergent fiber, Contraception in Andean bears has included melenges-
crude protein, soluble sugars, and crude fat were 12.8%, trol acetate (MGA implants), hysterectomy, medroxypro-
70.0%, 80.3%, and 64.5%, respectively. Acid detergent gesterone injections (Depo-Provera), and deslorelin acetate
fiber showed zero digestibility, and the mean transit time of (Suprelorin) (Association of Zoos & Aquariums RMC
food through the gastrointestinal tract was between 8 and [Reproductive Management Center] at the Saint Louis
24 hours. Results indicated that Andean bears have limited Zoo). MGA implants are no longer recommended for
capabilities to digest higher fiber diets.11 carnivores generally, and in captive Andean bears its use has
been reduced considerably since 2005 (M Agnew, personal
Behavior and Related Issues communication, March 09, 2017). MGA implants have
been used in Colombian Andean bears, however, without
As in other bear species, stereotypies in captivity are common complications (F Nassar-Montoya, personal communica-
in Andean bears and require investigation to determine the tion, August 10, 2016). Reproductive issues reported in
underlying causes. In one facility, two captive Andean bears Andean bears include polycystic ovaries,23 recurrent vagini-
spent as much as 80%–85% of observed time engaging in tis due to Escherichia coli,17 and endometrial hyperplasia.24
stereotypical movements. The enclosure, husbandry, and Recently, molecular sexing using polymerase chain reaction
medical issues were demonstrated to be associated with the has been used on fecal samples found in the bear’s natural
stereotypies, and changing the habitat to a more naturalistic environment to determine the animal’s gender, a useful tool
environment provided mental stimulation for the bears, and in monitoring populations.25
reduced the stereotypies significantly, improving chronic
dental pathologies like alveolar injuries and canine fractures
due to the chewing of the bars that have been described Handling, Restraint, and Chemical
in other species.12 Further investigation is needed with Immobilization
regard to how disease, perception and expression of pain,
husbandry, and appropriate environmental stimulation can With cubs weighing less than 25 kg, manual physical
affect captive Andean bear welfare both negatively and posi- restraint is possible using nets, heavy gloves, and blankets.6,26
tively.13,14 Complementary therapies, such as Bach flower As with other bears, manual restraint should not be used
remedies in conjunction with an environmental enrichment with larger animals. Operant conditioning in captive Andean
program, were used at one facility and resulted in a decrease bears facilitates handling, training for direct drug admin-
in abnormal, and an increase in natural, behaviors.15 istration, oral drug ingestion, dental health assessment,
blood sampling, nail trimming, radiographs, endoscopy,
Reproduction and Related Medical Issues and other procedures.27 Nevertheless, even with training,
protected contact should be used with Andean bears, and
Captive Andean bears have been classified as polyestrous, the benefits of squeeze cages and pole syringe should not
facultative seasonal breeders,5 showing follicular and luteal be underestimated. The same safe practices and precautions
phases of 8 days and 22 days, respectively. Females usually used in other bear species should be utilized with Andean
have three to four ovarian cycles per year.16 Some authors bears. Similarly, the complications of anesthesia that occur
report a 7-month gestation period,17 while others believe in other bear species can occur with this species as well,
that embryonic diapause (delayed implantation) makes necessitating an appropriate anesthesia plan, whether in a
gestation length variable and difficult to calculate.5,18 One zoological setting or in the field.
to two cubs (and up to three cubs in North American In Latin American zoos and in field research on Andean
zoos)19 are born to females year round in zoos that lie bears, the standard anesthetic protocols have included
within the bears’ natural distribution range.5 In facilities ketamine-xylazine (KX) and/or zolazepam-tiletamine
in the northern hemisphere—and thus outside the species’ (ZT). The choices of these drugs has been due primarily
distribution—Andean bears have a tendency toward a to poor or nonavailability of other alpha-2 agonists, ben-
winter birthing season.5,17 zodiazepines, and potent opioids in some range countries.
Sexual maturity in captivity ranges from 3 to 7 years ZT has been used in Andean bears at 3.2–11.1 mg/kg.28
with a mean range of 4 years for females and 5 years for Combinations of ketamine (2.5–4 mg/kg), medetomidine
males.5,20 Reproductive life spans in captive individuals (0.035–0.075 mg/kg), and midazolam (0.05–0.09 mg/kg)
are around 15–17 years for females but almost double have also been successfully utilized for spectacled bears. An
for males.20 Assisted reproductive techniques in Andean ongoing, prospective study on Andean bear medicine and
bears, first attempted in the late 1990s, consisted mainly of anesthesia is investigating additional drugs and dosages in
electroejaculation and cryopreservation of semen.21,22 More Andean bears using combination protocols with ketamine,
recently, semen collection via urethral catheterization has midazolam, and dexmedetomidine or medetomidine.
550 SE C T I O N 15 Carnivores
TABLE
78.1 Anesthetic Protocols in Andean Bears (Tremarctos ornatus) used at Bioparque Wakatá Colombia
Reversal Agent/Dose
Drug Dose (mg/kg), IM mg/kg Comments n*
Ketamine 3.6–9 Yohimbine 0.11 IV Animals required redosing of ketamine 18
Xylazine 0.5–1.5
Ketamine 5.6–7.3 Atipamezole 0.2 IV Large volume of dexmedetomidine could be 5
Dexmedetomidine 0.02–0.035 Flumazenil 0.01 IV problematic with darts. Animals needed
Midazolam (KDM) 0.27–1.01 redosing of KDM
Tiletamine- 6–6.5 Flumazenil 0.01 IV Prolonged recovery 6
zolazepam
Ketamine 2–6.2
Ketamine 4 Atipamezole 0.24 IV Initial effects within 2–5 minutes. No redosing 2
Medetomidine 0.04 Flumazenil 0.01 IV needed
Midazolam 0.1
*n = Number of animals.
TABLE
78.3 Hematologic Reference Values for Andean Bears (Tremarctos ornatus)
Colombia* ZIMS†
*The data were obtained from 15 captive bears, from two different institutions: Bioparque Wakatá and Zoologico de Cali. Some of these data contribute to the
ZIMS data base as well.
†
Teare JA, editor: “Tremarctos ornatus No selection by gender All ages combined Standard International Units 2013 CD.html” in Species360 Physiological
Reference Intervals for Captive Wildlife: A CD-ROM Resource, Species360, Bloomington, MN, 2013.
TABLE
78.4 Serum Chemistry Reference Values for Andean Bears (Tremarctos ornatus)
Colombia* ZIMS†
*The data was obtained from 9 captive bears, from the institution Bioparque Wakatá. Some of this data contributes to the ZIMS data base.
†
Teare, J.A. (ed.): 2013, “Tremarctos ornatus No selection by gender All ages combined Standard International Units 2013 CD.html” in Species360 Physiological
Reference Intervals for Captive Wildlife: A CD-ROM Resource., Species360, Bloomington, MN.
552 SE C T I O N 15 Carnivores
A B
• Figure 78.1 Osteoarthritis in (A) elbow and (B) knee of a captive Andean bear (Tremarctos ornatus)
older than 20 years.
and giant cell infiltrates.33,36 No clear conclusions regarding Dr. Diego X. Medina, Dr. Fernando Nassar-Montoya, Dr.
diagnosis or treatment are known either, although oclaci- José Orlando Feliciano, Dr. Diego Soler-Tovar, Catalina
tinib maleate, an inhibitor of pruritogenic, proinflammatory Rodríguez, Juliana Osorio, Dr. Juliana Peña, Dr. Juan C.
cytokines used to treat pruritic allergic dogs, gave promising Rozo, Dr. Rafael Torres, Parque Jaime Duque, Fundación
results in three affected Andean bears.36 Further information Nuevos Horizontes and ALVEFAS, A.C.
about the efficacy of this drug in treating alopecia syndrome
in Andean bears is still needed, however.
In captive Andean bears, multiple types of neoplasia have References
been reported. These include lymphosarcoma,38 gingival 1. Del Moral JF, Lameda FI: Registros de ocurrencia del oso andino
epidermoid carcinoma of the mandibular bone,17 meso- (Tremarctos ornatus Cuvier, 1825) en sus límites de distribución
thelioma, thymoma, cardiac rhabdomyosarcoma, squamous nororiental y austral, RevMus Argent Cienc Nat 13(1):7–19,
cell carcinoma, pyloric leiomyoma, mammary adenoma, 2011.
spindle cell thymoma, cholangiosarcoma, and transitional 2. Kattan G, Hernández OL, Goldstein I, et al: Range fragmenta-
cell carcinoma of the urinary bladder.29,30 tion of the Spectacled Bear Tremarctos ornatus in the northern
In studies on the parasites of free-ranging Andean bears Andes, Oryx 38(2):1–10, 2004.
in Peru, Blastocystis sp., Giardia sp.,17,39 and Cryptosporidium 3. Ruíz-García M: Molecular population genetic analysis of the
sp. were found. All three of these parasites may reflect inter- Spectacled Bear, Hereditas 138(2):81–93, 2003.
4. Ministerio del Medio Ambiente: Programa Nacional Para la
actions between domestic animals, human settlements, and
Conservación en Colombia del Oso Andino (Tremarctos ornatus),
wildlife as well as the presence of polluted water sources. Bogotá, 2001, Imprenta Nacional.
In one study, rhabditid and ascarid nematodes were also 5. García-Rangel S: Andean bear Tremarctos ornatus natural history
identified in wild Andean bears.39 Preliminary data of a and conservation, Mammal Rev 42(2):85–119, 2012.
serologic screening survey on captive Andean bears in 6. Collins D: Ursidae. In Miller RE, Fowler ME, editors: Fowler’s
Colombia found 11 of 13 individuals (84.6%) positive for zoo and wild animal medicine (vol 8). St. Louis, MO, 2015,
Toxoplasma gondii antibodies using the latex agglutination Elsevier, pp 498–508.
test.38 Canine distemper has caused perinatal infection in 7. Figueroa J: Revisión de la dieta del oso andino Tremarctos ornatus
captive Andean bears29,40; but whether other potential viral (Carnivora: Ursidae) en América del Sur y nuevos registros para
agents such as herpesvirus type-I and canine adenovirus el Perú. Rev. Mus. Argentino Cienc, Nature 15(1):1–27, 2013.
type-I affect Andean bears remains unknown.6,41 8. Lisi KJ, Barnes TL, Edwards MS: Bear weight management:
a diet reduction plan for an obese spectacled bear (Tremarctos
In Colombia, the habitat of wild Andean bears over-
ornatus), J Zoo Aquar Res 1(2):81–84, 2013.
laps with the territories of domestic and invasive species. 9. Gerstner K, Liesegang A, Hatt JM, et al: Seasonal body mass
Infectious diseases such as leptospirosis and rabies as well changes and feed intake in spectacled bears (Tremarctos ornatus)
as parasites contracted from these species are a growing at Zurich Zoological Garden, J Zoo Aquar Res 4(3):121–126,
problem for the bears. In the wild, dental disease has been 2016.
reported in juvenile bears, as well as decreased growth in 10. Paisley L, Garshelis DL: Activity patterns and time budgets of
females and even infanticide (JO Feliciano, personal com- Andean bears (Tremarctos ornatus) in the Apolobamba Range of
munication, March 12, 2017). Bolivia, J Zool 268(1):25–34, 2005.
11. Goldman I, Silver SC, Dierenfeld ES: Passage and digestion in
the Spectacled Bear (Tremarctos ornatus) fed a zoo-based diet
Conclusions moderately high in fiber. In Proceedings of the Nutrition Advi-
sory Group - 4th Conference on Zoo and Wildlife Nutrition.
Andean bear health both in captivity and in the wild remains
Disney Animal Kingdom, 2001, 4:92.
a vast opportunity for research. Much of the information 12. Wenker CJ, Stich H, Müller M, et al: Retrospective study of
regarding the bears biological and medical aspects has been dental conditions of captive brown bears (Ursus arctos spp.) com-
collected from zoos in the Northern hemisphere, outside of pared with free-ranging Alaskan grizzlies (Ursus arctos horribilis),
their natural range. Zoological collections, rescue centers, J Zoo Wildl Med 30(2):208–221, 1999.
and wildlife stations within the Andean bears’ geographic 13. Maslak R, Agnieszka S, Hill SP: Some aspects of locomotory
distribution are urged to report and publish their find- stereotypies in spectacled bears (Tremarctos ornatus) and changes
ings on this increasingly endangered animal; research on in behavior after relocation and dental treatment, J Vet Behav
nutrition, anesthesia, infectious and noninfectious diseases, 8:335–341, 2013.
reproduction, clinical medicine, and pharmacology are 14. Renner MJ, Lussier PJ: Environmental enrichment for the
clearly needed for Andean bears, both those living in captive captive spectacled bear (Tremarctos ornatus), Pharmacol Biochem
Behav 73:279–283, 2002.
environments and to help conserve the populations remain-
15. Puentes K, Rodríguez-Álvarez C, Cardona M: Evaluación del
ing in the wild. uso de esencias florales para el manejo comportamental de un
oso de anteojos (Tremarctos ornatus) en el Bioparque Wakatá,
Acknowledgments Revista de Conservación y Bienestar Animal Número 1:2016. From:
www.parquejaimeduque.com. (Accessed 14 February 2017).
Special thanks to Dr. Ellen Wiedner, Dr. Eduardo Valdes, Dr. 16. Enciso MA, Caracterização da atividade ovariana no Urso-
Anneke Moresco, Dr. Mary Agnew, Dr. Roberto Fecchio, de-óculos (Tremarctos ornatus Cuvier, 1825) mediante análise
554 SE C T I O N 15 Carnivores
de metabólitos fecais de esteróides sexuais. Retrieved from 30. Murray S, Sanchez CD, Siemering GH, et al: Transitional cell
www.teses.usp.br/teses/ (Accessed 4 January 2017), 2013. carcinoma of the urinary bladder in a spectacled bear (Tremarctos
17. Von Hohendorff R, Giacomini C: Carnivora - Ursidae (Ursos). ornatus), Vet Rec 158:306–307, 2006.
In Cubas ZS, Ramos JC, Catão-Dias JL, editors: Tratado de 31. Diaz-Parra H: Espondilosis en Oso de Anteojos (Tremarctos
Animais Selvagens: medicina veterinária, ed 2, Sao Paulo, 2014, ornatus) y gota viceral en Cóndor de los Andes (Vulture gryphus):
Roca, pp 880–892. Casos observados en la Fundación zoológico Santacruz, Mem
18. Garshelis DL: Family ursidae. In Wilson E, Mittermeier R, Conf Interna Med Aprovech Fauna Silv Exót Conv 2:37, 2016.
editors: Handbook of the mammals of the world: carnivores, Bar- 32. Vásquez V, Lara EL, Arias-Bernal L, et al: Diagnóstico de las
celona, Spain, 2009, Lynx Edicions, Conservation International enfermedades peridontales en un grupo de carnívoros nativos en
and IUCN, pp 448–497. cautiverio en el Zoológico Jaime Duque. Revista de Medicina
19. Peyton B: Spectacled bear conservation action plan. In IUCN/ Veterinaria, Universidad de La Salle 11:43–57, 2006.
SSC Bear and Polar Bear Specialist Group, editor: Bears: Status, 33. J€ager K, Langguth S, Einspanier A, et al: The alopecia-syndrome
Survey and Conservation Action Plan, Switzerland, 1999, Gland. of Spectacled bears (Tremarctos ornatus). What do we know, what
20. Rodríguez-Clark KM, Sánchez-Mercado AY: Population manage- can we do?, J Comp Pathol 148:90, 2013.
ment of threatened taxa in captivity within their natural ranges: 34. Owen M, Shanks J, Sutherland-Smith M, et al: Update on an
Lessons from Andean bears (Tremarctos ornatus) in Venezuela, investigation into a chronic skin disorder among captive Andean
Biol Conserv 129:134–148, 2006. Bears in North America, International Bear News 18(2):25–26,
21. Sanchez R, Nassar-Montoya F: Estudio reproductivo y congel- 2009.
ación de semen del Oso de Anteojos (Tremarctos ornatus), Rev 35. Kolter L, Bechstein N, Schachtner M, et al: Adding stones to the
Med Vet (B Aires) 1(1):25–33, 1997. puzzle : approaches to the alopecia syndrome in captive Andean
22. Enciso M, Bermúdez L, Evangelista S, et al: Estudio preliminar bears (Tremarctos ornatus), International Bear News 23(3):20–21,
de colección de semen en oso de anteojos (Tremarctos ornatus), 2014.
Rev Inv Vet Perú 17(1):77–80, 2006. 36. Leclerc A, Lécu A, Bechstein N, et al: Spectacled Bear (Tremarctos
23. Nassar-Montoya F, Sanchez R, Vega LA, et al: Estudio Clínico del ornatus) Alopecia syndrome: An update. Proceedings of the
Oso de Anteojos (Tremarctos ornatus), Bogotá, Colombia, 1996, Annual Conference of the American Association of Zoo Veteri-
Icono, pp 23–35. narians, 2015:126–127.
24. Munson L, Moresco A, Calle PP: Adverse effects of contracep- 37. Melo S, Vargas CE, Arias-Bernal L, et al: Evaluación de un
tives in captive and free-ranging wildlife. In Asa CS, Porton IJ, protocolo de medicina biológica para el tratamiento del sín-
editors: Wildlife contraception: issues, methods, and applications, ed drome dermatológico del oso andino (Tremarctos ornatus) en el
1, Baltimore, MD, 2005, The Johns Hopkins University Press. Zoológico Jaime Duque. La Siringe. Boletin de la facultad de
25. Caselli C, Maturrano L: Sexaje Molecular a Partir de Heces medicina veterinaria y de zootecnia, Universidad Antonio Nariño
en Osos de Anteojos (Tremarctos ornatus), Rev Inv Vet Perú 6(6):32–38, 2014.
27(2):252–258, 2016. 38. Arias-Bernal L, Yarto E: Unpublished manuscript. 2016.
26. Fowler ME: Carnivores. In Fowler ME, editor: Restraint and 39. Figueroa J: New records of parasites in free-ranging Andean bears
handling of wild and domestic animals, ed 2, Ames, IA, 1995, from Peru, Ursus 26(1):21–27, 2015.
Iowa State Press, pp 221–235. 40. Schonbauer M, Kolbl S, Schonbauer-Langle A: Pertinatale stau-
27. Otaki Y, Kido N, Omiya T, et al: A new voluntary blood collec- peinfektion bei drei eisbaren (Ursus maritimus) und bei einem
tion method for the Andean Bear (Tremarctos ornatus) and Asiatic brillenbaren (Tremarctos ornatus), Verh Int Symp Erkrank Zoot
Black Bear (Ursus thibetanus), Zoo Biol 34:497–500, 2015. (26):131–136, 1984.
28. Caulkett N, Fahlman A: Ursids (Bears). In West G, Heard D, 41. Beineke A, Baumgärtner W, Wohlsein P: Cross-species trans-
Caulkett N, editors: Zoo animal and wildlife immobilization and mission of canine distemper virus—an update, One Health
anesthesia, ed 2, Oxford, 2014, Wiley-Blackwell, pp 599–606. (1):49–59, 2015.
29. Bourne DC, Cracknell JM, Bacon HJ: Veterinary issues related
to bears (Ursidae), Int Zoo Yearb 44:16–32, 2010.
79
Canine Distemper Vaccination in
Nondomestic Carnivores
TIMOTHY A. GEOROFF
C
anine distemper virus (CDV), a member of Morbil- to control disease; however, the use of MLV CDV vac-
livirus genus in the family Paramyxoviridae, is a cines has generally not been recommended in nondomestic
significant viral disease of both captive and free- carnivores because of the potential reversal of attenuation
ranging carnivores globally.1 Morbidity and mortalities and vaccine-induced CDV (Table 79.1). Inactivated CDV
due to CDV infection have been described in nearly all whole-virus vaccines, however, do not produce sufficient
extant families of Carnivora (Ailuridae, Canidae, Felidae, immunity to prevent infection after virus challenge (sterile
Hyaenidae, Mephitidae, Mustelidae, Otariidae, Phocidae, immunity) and are no longer available commercially in the
Procyonidae, Ursidae, and Viverridae)1–7 including large- United States.12 Recombinant subunit CDV vaccines have
scale epidemics in multiple species.6,8,9 Disease due to become popular in zoological medicine because of their
CDV continues to remain an issue in both captive and safety profile; however, availability and questions regard-
free-ranging populations due to CDV persistence through ing the ability of some products to adequately elicit an
a wide range of carnivore reservoir hosts.10–13 immune response and provide adequate protection against
CDV is an RNA virus that contains, similar to other CDV remain issues. This chapter summarizes the current
paramyxoviruses, six structural proteins: nucleocapsid (N), knowledge on canine distemper vaccination with special
phosphoprotein (P), large protein (L), single-envelope- interest in the application in nondomestic carnivores (see
associated or matrix protein (M), two glycoproteins— Chapter 44).
hemagglutinin (H) and fusion (F) protein, and two accessory
nonstructural proteins (C and V).14 CDV is primarily trans-
mitted through respiratory secretions although transmission Recombinant Viral Vector Vaccines
through other body excretions such as urine can result in
infection in susceptible animals. CDV exhibits lympho-, Recombinant viral vector vaccines are genetically engineered
neuro-, and epitheliotropism and disease most commonly vaccines produced through recombinant DNA technology
involves the respiratory, enteric, integumentary, and central that involve insertion of DNA encoding key antigens into
nervous system symptoms. Secondary infections reflecting a different viral vector (poxvirus, adenovirus, or alphavirus)
virus-induced immunosuppression are commonly seen with for delivery. These vaccines have a similar safety profile to
CDV and may complicate clinical disease.15,16 Clinical signs inactivated (killed) subunit vaccines but express antigens
and the severity of disease are affected by various factors inside vector infected cells so that MHC class I and class II
including host age and immune status, species sensitivity presentation can occur efficiently and stimulate both cell-
to virus, and viral strain virulence, as well as other environ- mediated and humoral immune responses. The expression
mental factors.1 of only recombinant proteins allows the targeting of the
There is currently no known effective treatment for clini- immune response against a few antigens produced by the
cal disease from CDV infection, and treatment is limited pathogen without including the entire pathogen, thereby
to supportive care and treatment of secondary infections. significantly reducing or eliminating the risk of vaccine-
Disease from CDV is best prevented and controlled through induced disease.
vaccination. There are two major categories of commercially Recombinant CDV (rCDV) vaccines have been created
available vaccines for CDV: modified-live virus (MLV) vac- that have CDV genes for the H and F proteins inserted
cines and canarypox vectored recombinant vaccines. The in a live canarypox viral vector. Adequate host immune
introduction of MLV vaccines in the 1950s has helped response against the H protein is considered important
555
556 SE C T I O N 15 Carnivores
TABLE
79.1 Reports by Species of Vaccine-Induced Canine Distemper
Tissue Culture
Species Vaccine Vaccine Type Vaccine Strain Origin Reference
Canidae
African Lycaon pictus Paramune 5* MLV combo (CDV, n/a n/a 17
wild CAV, CPIV) + Lepto
dog bacterin vaccine
Candur SHLP† MLV CDV + killed Rockborn Canine kidney 18
CPV, CAV-1 + Lepto
bacterin vaccine
Bush dog Speothos venaticus DA2MP‡ MLV combo vaccine Snyder Hill Canine kidney 19
(CDV, CAV-2, CPIV,
CPV)
Gray fox Urocyon Fromm-D§ MLV CDV vaccine Onderstepoort Chicken embryo 20
cinereoargenteus Vanguard D¶ MLV CDV vaccine Snyder Hill Canine kidney 20
Tissuevax** MLV CDV vaccine Rockborn Canine kidney 20
Maned Chrysocyon n/a MLV CDV vaccine n/a n/a 21
wolf brachyurus
Ailuridae
Red Ailurus fulgens Epivax-TC- MLV CDV vaccine n/a Chicken embryo 22
panda plus††
Fervac-D‡‡ MLV CDV vaccine Lederle Chicken embryo 24
Fromm-D§ MLV CDV vaccine Onderstepoort Chicken embryo 23
Procyonidae
Kinkajou Potos flavus Vanguard¶ MLV combo vaccine Rockborn Canine kidney 25
(CDV, CAV-2)
Mustelidae
Black- Mustela nigripes n/a MLV CDV vaccine n/a Chicken embryo 26
footed
ferret
European Mustela lutreola Galaxy MLV combo (CDV, Onderstepoort Primate Vero 27
mink 6-MPH-L§§ CAV-2, CPIV, and
CPV) + Lepto
bacterin vaccine
and may prevent subsequent CDV infection.28 A vaccinia shedding.30 Recombinant CDV vaccines have also been
vectored vaccine using insertion of genes encoding the H shown to be safe to administer during pregnancy.31 Adverse
and F proteins from measles virus, another morbillivirus, reactions historically seen with MLV CDV vaccines such
has been demonstrated to produce neutralizing antibod- as postvaccinal encephalitis are not possible with rCDV
ies to CDV to protect against viral challenge in domestic vaccines.30 Recombinant CDV vaccines have been shown to
dogs.29 The canarypox viral vector used in the CDV vaccine successfully immunize domestic ferrets and protect against
has the advantage that the canarypox vector is replication- subsequent viral challenge.32
incompetent in mammalian hosts so the vaccine is inca- A commercially available monovalent canarypox-
pable of reversion to virulence and does not result in viral vectored rCDV vaccine, PUREVAX ferret distemper
CHAPTER 79 Canine Distemper Vaccination in Nondomestic Carnivores 557
(Merial, Athens, Georgia, USA) approved for use in the seroconvert postvaccination.36 Higher doses of vaccine
United States in domestic ferrets, was first made available (or increased concentration of antigen) appear necessary
in 2001. This vaccine has been safely utilized via extra-label to elicit an adequate protective immune response when
use in numerous species of nondomestic carnivores, and administering rCDV vaccine orally.44 Because of this,
the humoral response to vaccination with this vaccine has parenteral administration of rCDV vaccines is recom-
been assessed in several species (Table 79.2). The ferret mended in captive nondomestic carnivores for routine
distemper rCDV vaccine has been advocated for use by preventative health programs.
many Association of Zoos & Aquariums (AZA) Species The author is unaware of any reports of natural disease
Survival Program veterinary advisors because of its safety from CDV subsequent to vaccination in nondomestic
profile and evidence of antibody response to vaccination carnivores vaccinated regularly (every 1–3 years) with
in multiple species studied, and has been widely utilized the ferret rCDV distemper vaccine. Recombinant CDV
for vaccination of nondomestic carnivores within North vaccines appear very safe with very few adverse effects
American zoos. This vaccine, however, has been frequently reported and no reports of vaccine-induced disease. There
on manufacturer back order and unavailable for periods of is one report of erythema multiforme in a spotted hyena
up to several years at a time, forcing veterinarians to search (Crocuta crocuta) occurring after vaccination with ferret
for other rCDV vaccine options. A similar recombinant rCDV vaccine.51 Following vaccination plus booster with
vaccine, RECOMBITEK canine distemper, is approved for the ferret rCDV vaccine, antibody levels considered protec-
use in domestic dogs and available in several combination tive (≥1:20 for vaccination response)12 against CDV have
preparations that include other MLV vaccine components been detected postvaccination in nondomestic Canidae,
for different canine infectious diseases depending on the Herpestidae, Mustelidae, Phocidae, and Ursidae (see Table
preparation. This vaccine more recently became available 79.2). Duration of humoral antibodies following an initial
in a monovalent version including only the rCDV vaccine series of rCDV vaccinations followed by a booster at 1
(RECOMBITEK CDV). These vaccines involve the same year in domestic dogs has been shown to persist for at
live canarypox viral vector with H and F protein gene inser- least 36 months.30 Antibody persistence up to and beyond
tions as the ferret distemper vaccine; however, the amount 1 year also appears to occur post-rCDV vaccination in
of vaccine antigen load differs between the ferret distemper some nondomestic species (see Table 79.2). In large felids
rCDV and canine rCDV vaccine series. The actual amounts (Panthera spp.) receiving two or more prior ferret rCDV
are proprietary, but the ferret distemper rCDV vaccine dose vaccinations that showed evidence of prior seroconversion,
is approximately eight times greater than the canine rCDV 91% (21/23) of animals evaluated maintained SN titers at
vaccine dose. This difference is based on the demonstrated ≥1:24 for at least 24 months postvaccination.52 Another
protective doses that were established in licensure studies for report indicates antibodies may not persist as long following
ferrets and domestic dogs (J Maki, personal communica- ferret distemper rCDV vaccination in other canid species.36
tion, March 29, 2017). Results of canine rCDV vaccination in nondomestic
Recombinant CDV vaccines typically induce lower species have been more varied. Several reports and addi-
serum-neutralizing (SN) titers compared to MLV CDV tional data indicate some of the canine rCDV vaccines
vaccines in nondomestic carnivores although stimula- have successfully induced seroconversion in nondomestic
tion of cell-mediated immune response may also be an canids,37,39 raccoons (Procyon lotor) (E Ramsey, personal
important factor in establishing protective immunity. communication, August 3, 2014), and red pandas (Ailurus
Recombinant CDV vaccines appear to require minimally fulgens) (A Guthrie, personal communication, April 30,
an initial vaccine plus booster to produce an SN antibody 2015). In a study in tigers, however, vaccination with one
response in nondomestic carnivores,37,49 although single of the rCDV combination vaccines produced only low titers
vaccination may induce seroconversion in some species.39 in 2/6 animals following initial vaccination plus booster.33
Recombinant CDV vaccines are superior to MLV CDV Disease due to circulating wild-type CDV has also been
vaccines in inducing a neutralizing antibody response documented in a snow leopard (Panthera uncia) despite
in the presence of maternal antibody; however, survival prior vaccination with the monovalent canine rCDV
was only 14% following challenge in ferrets vaccinated vaccine 3 months prior.33a The difference in vaccine antigen
parenterally with rCDV vaccine in the presence of mater- loads may have had some impact on these results and high-
nal antibody.32 It is therefore recommended to vaccinate lights the need to determine species-specific vaccination
multiple times for an initial series in naïve animals less recommendations. It may be possible to overcome some
than 16 weeks of age. The American Animal Hospital of these challenges by administering increased volume of
Association (AAHA) recommends a series of vaccinations vaccine (>1 mL), although this is unclear based on current
every 3–4 weeks between the ages of 6 and 16 weeks fol- information and further study is needed. There are also
lowed by a booster at 12 months and then revaccination unanswered questions as to the safety of vaccinating nondo-
every ≥3 years with either rCDV or MLV CDV vaccines mestic carnivores with polyvalent vaccines containing other
for domestic dogs.50 Route of rCDV vaccine administra- MLV vaccine components (see Chapter 57 by Lamberski
tion also appears significant. African wild dogs (Lycaon in Fowler’s Zoo and Wild Animal Medicine Current Therapy
pictus) vaccinated orally with rCDV vaccine failed to [vol 7]).
TABLE
79.2 Summary of Canine Distemper Vaccine Studies in Nondomestic Carnivores
Continued
CHAPTER 79 Canine Distemper Vaccination in Nondomestic Carnivores
559
TABLE
79.2 Summary of Canine Distemper Vaccine Studies in Nondomestic Carnivores—cont’d
8. Roelke-Parker ME, Munson L, Packer C, et al: A canine dis- 28. von Messling V, Zimmer G, Herrler G, et al: The hemagglutinin
temper virus epidemic in Serengeti lions (Panthera leo), Nature of canine distemper virus determines tropism and cytopathoge-
379:441–445, 1996. nicity, J Virol 75:6418–6427, 2001.
9. van de Bildt MW, Kuiken T, Visee AM, et al: Distemper outbreak 29. Taylor JS, Pincus J, Tartaglia C, et al: Vaccinia virus recombinants
and its effect on African wild dog conservation, Emerg Infect Dis expressing either the measles virus fusion or hemagglutinin gly-
8:211–213, 2002. coprotein protect dogs against canine distemper virus challenge,
10. Almberg ES, Cross PC, Smith DW: Persistence of canine dis- J Virol 65:4263–4274, 1991.
temper virus in the Greater Yellowstone Ecosystem’s carnivore 30. Larson LJ, Schultz RD: Three-year duration of immunity in
community, Ecol Appl 20:2058–2074, 2010. dogs vaccinated with a canarypox-vectored recombinant canine
11. Gilbert M, Soutyrina S, Seryodkin I, et al: Canine distemper distemper virus vaccine, Vet Ther 8:101–106, 2007.
virus as a threat to wild tigers in Russia and across their range, 31. Wimsatt J, Biggins DE, Williams ES: The quest for a safe and
Integr Zool 10:329–343, 2015. effective canine distemper virus vaccine for black-footed ferrets.
12. Greene CE, Vandevelde M: Canine distemper. In Greene CE, In Roelle JE, Miller BJ, Godbey JL, et al, editors: Recovery of
editor: Infectious diseases of the dog and cat, ed 4, St. Louis, MO, the black-footed ferret: progress and continuing challenges, Fort
2012, Elsevier, pp 25–42. Collins, 2006, Proc Symp Status Black-Footed Ferret Habitat,
13. Trebbien R, Chriel M, Struve T, et al: Wildlife reservoirs of pp 248–266.
canine distemper virus resulted in a major outbreak in Danish 32. Welter J, Taylor J, Tartaglia J, et al: Vaccination against canine
farmed mink (Neovison vison), PLoS ONE 9:e85598, 2014. distemper virus infection in infant ferrets with and without
14. Martella V, Elia G, Buonavoglia C: Canine distemper virus, maternal antibody protection, using recombinant attenuated
Vet Clin North Am Small Anim Pract 38:787–797, 2008. poxvirus vaccines, J Virol 74:6358–6367, 2000.
15. Lemberger KY, Gondim LF, Pessier AP, et al: Neospora caninum 33. Sadler RA, Ramsay E, McAloose D, et al: Evaluation of two
infection in a free-ranging raccoon (Procyon lotor) with concur- canine distemper virus vaccines in captive tigers (Panthera tigris),
rent canine distemper virus infection, J Parasitol 91:960–961, J Zoo Wildl Med 47:558–563, 2016.
2005. 33a. Chinnadurai SK, Kinsel MJ, Adkesson MJ, et al: Canine
16. Silinski S, Robert N, Walzer C: Canine distemper and toxoplas- distemper in a vaccination snow leopard (Panthera uncia), J Zoo
mosis in a captive snow leopard (Uncia uncia) – a diagnostic Wildl Med 48:1200–1203, 2017.
dilemma, Verh Ber Zootiere 41:107–111, 2003. 34. Kock R, Chalmers WS, Mwanzia J, et al: Canine distemper
17. McCormick AE: Canine distemper in African cape hunting antibodies in lions of the Masai Mara, Vet Rec 142:662–665,
dogs (Lycaon pictus) possibly vaccine induced, J Zoo Anim Med 1998.
14:66–71, 1983. 35. Coke RL, Backues KA, Hoover JP, et al: Serologic responses after
18. Durchfeld B, Baumgärtner W, Herbst W, et al: Vaccine-associated vaccination of fennec foxes (Vulpes zerda) and meerkats (Suricata
canine distemper infection in a litter of African hunting dogs suricatta) with a live, canarypox-vectored canine distemper virus
(Lycaon pictus), Zentralbl Veterinarmed B 37:203–212, 1990. vaccine, J Zoo Wildl Med 36:326–330, 2005.
19. McInnes EF, Burroughs RE, Duncan NM: Possible vaccine- 36. Connolly M, Thomas P, Woodroffe R, et al: Comparison of oral
induced canine distemper in a South American bush dog and intramuscular recombinant canine distemper vaccination in
(Speothos venaticus), J Wildl Dis 28:614–617, 1992. African wild dogs (Lycaon pictus), J Zoo Wildl Med 44:882–888,
20. Halbrooks RD, Swango LJ, Schnurrenberger PR, et al: Response 2013.
of gray foxes to modified live-virus canine distemper vaccines, 37. Hidalgo E, Ortiz C, Mathieu C, et al: Serological response
J Am Vet Med Assoc 179:1170–1174, 1981. in red fox (Vulpes vulpes) to the canine distemper vaccination
21. Thomas-Baker B: Vaccination-induced distemper in maned with RECOMBITEK C6®. In Proceedings of the American
wolves, vaccination-induced corneal opacity in a maned wolf. In Association of Zoo Veterinarians, 2014.
Proceedings. Am Assoc Zoo Vet, 1985, p 53. 38. Vickers TW, Garcelon DK, Fritcher DL, et al: Evaluation of
22. Bush M, Montali RJ, Brownstein D, et al: Vaccine-induced canine an oral and food-based canarypox-vectored canine distemper
distemper in a lesser panda, J Am Vet Med Assoc 169:959–960, vaccine in endangered Channel Island foxes (Urocyon littoralis).
1976. In Proceedings of the American Association of Zoo Veterinarians,
23. Montali RJ, Tell L, Bush M, et al: Vaccination against canine 2004, pp 87–88.
distemper in exotic carnivores: successes and failures. In Proceed- 39. Bailey KL, Tritsch SS, Washburn TC, et al: Safety and immuno-
ings of the American Association of Zoo Veterinarians, 1994, pp genicity of vaccination of gray foxes (Urocyon cinereoargenteus)
340–344. with a multivalent recombinant canine distemper vaccine and
24. Bronson E, Denver MC, Karim B, et al: Vaccine-induced canine modified live parvovirus vaccine. In Proceedings of the American
distemper virus infection in a lesser red panda (Ailurus fulgens Association of Zoo Veterinarians, 2015, p 133.
fulgens) three years following last vaccination. In Proceedings 40. Harrenstien LA, Munson L, Ramsay EC, et al: Antibody
of the American Association of Zoo Veterinarians, 2003, pp responses of red wolves to canine distemper virus and
185–186. canine parvovirus vaccination, J Wildl Dis 33:600–605,
25. Kazacos KR, Thacker HL, Shivaprasad HL, et al: Vaccination- 1997.
induced distemper in kinkajous, J Am Vet Med Assoc 179: 41. Anderson K, Case A, Woodie K, et al: Duration of immunity in
1166–1169, 1981. red wolves (Canis rufus) following vaccination with a modified
26. Carpenter JW, Appel MJ, Erickson RC, et al: Fatal vaccine- live parvovirus and canine distemper vaccine, J Zoo Wildl Med
induced canine distemper virus infection in black-footed ferrets, 45:550–554, 2014.
J Am Vet Med Assoc 169:961–964, 1976. 42. Bronson E, Deem SL, Sanchez C, et al: Serologic response to a
27. Sutherland-Smith MR, Rideout BA, Mikolon AB, et al: Vaccine- canarypox-vectored canine distemper virus vaccine in the giant
induced canine distemper in European mink, Mustela lutreola, panda (Ailuropoda melanoleuca), J Zoo Wildl Med 38:363–366,
J Zoo Wildl Med 28:312–318, 1997. 2007.
CHAPTER 79 Canine Distemper Vaccination in Nondomestic Carnivores 563
43. Paré JA, Barker IK, Crawshaw GJ, et al: Humoral response and 50. Welborn LV, DeVries JG, Ford R, et al: 2011 AAHA canine
protection from experimental challenge following vaccination vaccination guidelines, J Am Anim Hosp Assoc 47:1–42, 2011.
of raccoon pups with a modified-live canine distemper virus 51. Hanley CS, Simmons HA, Wallace RS, et al: Erythema mul-
vaccine, J Wildl Dis 35:430–439, 1999. tiforme in a spotted hyena (Crocuta crocuta), J Zoo Wildl Med
44. Wimsatt J, Biggins D, Innes K, et al: Evaluation of oral and 36:515–519, 2005.
subcutaneous delivery of an experimental canarypox recombi- 52. Georoff TA: unpublished data, 2017.
nant canine distemper vaccine in the Siberian polecat (Mustela 53. van Heerden J, Bingham J, van Vuuren M, et al: Clinical and
eversmanni), J Zoo Wildl Med 34:25–35, 2003. serological response of wild dogs (Lycaon pictus) to vaccination
45. Peper ST, Peper RL, Mitcheltree DH, et al: Utility of two against canine distemper, canine parvovirus infection and rabies,
modified-live virus canine distemper vaccines in wild-caught J S Afr Vet Assoc 73:8–12, 2002.
fishers (Martes pennanti), Vet Q 36:197–202, 2016. 54. Jensen TH, Nielsen L, Aasted B, et al: Canine distemper virus
46. Peper ST, Peper RL, Kollias GV, et al: Efficacy of two canine dis- DNA vaccination of mink can overcome interference by maternal
temper vaccines in wild Nearctic river otters (Lontra canadensis), antibodies, Vaccine 33:1375–1381, 2015.
J Zoo Wildl Med 45:520–526, 2014. 55. Silin D, Lyubomska O, Ludlow M, et al: Development of a
47. Jessup DA, Murray MJ, Casper DR, et al: Canine distemper challenge-protective vaccine concept by modification of the viral
vaccination is a safe and useful preventive procedure for southern RNA-dependent RNA polymerase of canine distemper virus, J
sea otters (Enhydra lutra nereis), J Zoo Wildl Med 40:705–710, Virol 81:13649–13658, 2007.
2009. 56. Kapil S, Allison RW, Johnston L, et al: Canine distemper virus
48. Goodrich JM, Williams ES, Buskirk SW: Effects of a modified- strains circulating among North American dogs, Clin Vaccine
live virus canine distemper vaccine on captive badgers (Taxidea Immunol 15:707–712, 2008.
taxus), J Wildl Dis 30:492–496, 1994.
49. Quinley N, Mazet JA, Rivera R, et al: Serologic response of
harbor seals (Phoca vitulina) to vaccination with a recombinant
canine distemper vaccine, J Wildl Dis 49:579–586, 2013.
SECTION 16
Great Apes
80 Infectious Diseases of Orangutans in their Home
Ranges and in Zoos, 565
564
80
Infectious Diseases of Orangutans in
their Home Ranges and in Zoos
JOOST PHILIPPA AND ROSALIE D ENCH
565
TABLE
80.1 Important Infectious Diseases of Orangutans (Pongo spp.)
566 SE C T I O N 16 Great Apes
Antigen/
Antibody/ Pathogen
Reaction Isolation Wild* Rehab† Zoo‡ Mode of Transmission References
Virus
Adenoviridae Adenovirus + + + + + Fecal-oral 4, 74
Papovaviridae Polyomavirus + + + + − Fecal-oral 5
Herpesviridae Herpes simplex viruses (1 and 2) + − − + + Body fluids, direct contact 6, 7
Varicella zoster virus + − − − + Body fluids, direct contact, aerosol 6, 8
Epstein-Barr virus/human herpesvirus 4 + − + + + Body fluids, direct contact, bite 4, 6
Orangutan lymphocryptus virus/Pongine + + − − + Body fluids, direct contact, bite 9, 10
herpes viruses/Epstein-Barr virus–like
Cytomegalovirus/chimp CMV + − − − + Body fluids, direct contact 6
Poxviridae Monkeypox virus + + − + + Direct contact 11, 15
Hepadnaviridae HBV + − − + + Body fluids, direct contact 6
Orangutan hepadnavirus + + + + − Body fluids, direct contact 7
Picornaviridae Coxsackie viruses + + + + + Fecal-oral 4, 12, 15
Polio virus + + − + + Fecal-oral, direct contact 8
HAV + − − + + Fecal-oral 6, 7
Encephalomyocarditis virus + + − − + Fecal-oral, transplacental 13, 14
Togaviridae Sindbis virus + − + − + Vector: Culex sp. and Culiseta sp. 4
Rubella virus + − − − + Aerosol, direct contact 15
Flaviviridae Dengue virus + − + + − Vector: Aedes spp. 4
Japanese encephalitis virus + − + + − Vector: Culex spp. 4
Tembusu virus + − + + − Vector: Culex spp. 4
Langat virus + − − + − Vector: Ixodes sp. 4
Zika virus + − + + − Vector: Aedes spp. 4
Chikungunya virus + − − − + Vector: Aedes spp. 16
Reoviridae Rotavirus SA11 + − + + − Fecal-oral, direct contact 4
Orthomyxoviridae Influenza A virus + − − + + Aerosol 3, 6
Influenza B virus + − − + + Aerosol 3, 15
Paramyxoviridae Parainfluenza viruses 1 and 2 + − − − + Aerosol 6
Parainfluenza virus 3 + − − + + Aerosol 4, 6
Measles virus + − − − + Aerosol 6
Mumps virus + − + + + Body fluids, direct contact, aerosol 4, 15
Respiratory syncytial virus + − + + + Aerosol 3, 4, 6
Pneumoviridae Human metapneumovirus + − − + − Aerosol, direct contact 3
Rhabdoviridae Rabies virus − + − + − Body fluids, direct contact, bite 17
Bunyaviridae Hanta virus + − − − + Body fluids, direct contact 18
Filoviridae Reston Ebola virus§ + − + − − Body fluids, direct contact 19
Retroviridae SFV + + + + + Body fluids, direct contact 4, 15, 20
SRV + − − + − Body fluids, direct contact 7
STLV-I + + − + − Body fluids, direct contact 20
HTLV type I + − − + − Body fluids, direct contact 7
Bacteria
Gram + Staphylococcus sp. − + + − + Commensal overgrowth 8, 21
Gram + Streptococcus sp. − + + − + Commensal overgrowth 21, 22
Gram + Dermatophilus congolensis + − − − + Direct contact 23
Gram − Escherichia coli − + − + + Body fluids, direct contact 4, 8, 22
Gram − Salmonella sp. − + − + + Fecal-oral, direct contact, indirect 7, 24–26
Gram − Shigella sp. − + − + + Fecal-oral, direct contact, indirect 25, 27
Gram − Campylobacter sp. − + − − + Fecal-oral, direct contact, indirect 8, 27
Gram − Burkholderia pseudomallei − + − + + Indirect 4, 28
Gram − Francisella tularensis − + − − + Ingestion, aerosol, direct contact, 29
vectors (multiple species)
Gram − Klebsiella sp. − + − + + Direct contact, indirect 4, 8
¶
Gram − Leptospira interrogans + − + + − Urine, indirect 4
Gram − Helicobacter − + − − + Saliva, fecal-oral 30
Misc Mycobacterium tuberculosis complex + + − + + Aerosol 7, 8, 31, 32
Misc Mycobacterium avium − + − + − Aerosol 4
Continued
CHAPTER 80 Infectious Diseases of Orangutans in their Home Ranges and in Zoos
567
TABLE
80.1 Important Infectious Diseases of Orangutans (Pongo spp.)—cont’d
Antigen/
Antibody/ Pathogen
Reaction Isolation Wild* Rehab† Zoo‡ Mode of Transmission References
568 SE C T I O N 16 Great Apes
Endoparasite (GI)
Amebae Entamoeba coli − + + + + Fecal-oral 7, 33, 34
Amebae Entamoeba hartmani − + + + − Fecal-oral 33, 34
Amebae Entamoeba histolytica − + + + + Fecal-oral 33, 34
Amebae Entamoeba sp. − + + + + Fecal-oral 34, 35
Amebae Endolimax nana − + + + + Fecal-oral 33, 34
Amebae Iodamoeba buetschelii − + + + + Fecal-oral 33, 34
Amebae Ballamuthia mandrillaris − + − − + Fecal-oral 36
Amebae Blastocystis hominis − + − − + Fecal-oral 33
Flagellates Giardia sp. − + + + + Fecal-oral 7, 33–35
Flagellates Chilomastix mesnelii − + + + − Fecal-oral 33
Flagellates Chilomastix sp. − + + + − Fecal-oral 33, 35
Flagellates Dientamoeba fragilis − + + − − Fecal-oral 33
Flagellates Trichomonas − + + + + Fecal-oral 7, 8, 33
Ciliates Balantidium coli − + + + + Fecal-oral 4, 7, 33–35
Coccidia Cryptosporidium sp. − + + + + Fecal-oral 33, 37
Coccidia Cyclospora sp. − + − + − Fecal-oral 7
Nematodes Ascaris sp. − + − + − Fecal-oral 7, 34, 35
Nematodes Ancylostoma duodenale − + − + − Fecal-oral 7, 33
Nematodes Baylisascaris procyonis − + − − + Fecal-oral, direct contact 38
Nematodes Strongylida sp. − + + + + Fecal-oral 33, 35
Nematodes Strongyloides stercoralis − + − − + Fecal-oral, direct contact 33
Nematodes Strongyloides fuelleborni − + + + − Fecal-oral 33
Nematodes Strongyloides sp. − + + + + Fecal-oral 4, 33–35
Nematodes Trichostrongylus sp. − + + + − Fecal-oral 33, 34
Nematodes Trichuris spp. − + + + + Fecal-oral 4, 33–35
Nematodes Enterobius sp. − + + + + Fecal-oral 4, 33–35
Nematodes Mammomonogamus laryngeus − + − + − Fecal-oral 33
Nematodes Mammomonogamus sp. − + + + − Fecal-oral 33, 35
Nematodes Oesophagostomum sp. − + + + + Fecal-oral 7, 33
Nematodes Pongobius spp. (hugoti, foitovae) − + + − − Fecal-oral 33
Nematodes Spirurida sp. − + + + − Fecal-oral 33, 35
Cestodes Hymenolepis sp. − + + − − Fecal-oral 34
Trematodes Dicrocoeliidae sp. − + + + − Fecal-oral 33, 35
Trematodes Platynosomum fastotum − + + + − Fecal-oral 7
Endoparasite (Blood)
Sporozoa Plasmodium cynomolgi − + + + − Vector: Anopheles spp. 39
Sporozoa Plasmodium pitheci − + + + − Vector: Anopheles spp. 40, 41
Sporozoa Plasmodium silvaticum − + + + − Vector: Anopheles spp. 40, 41
Sporozoa Plasmodium vivax − + + + − Vector: Anopheles spp. 39
Sporozoa Plasmodium falciparum − + + + − Vector: Anopheles spp. 41
Sporozoa Plasmodium sp. − + + + − Vector: Anopheles spp. 4
Zoomastigophora Trypanosoma sp. − + − − + Vector: Triatominae 8
Fungi and Yeast
Yeast Candida sp. − + − − + Direct contact, indirect 8
Fungus Microsporum gypseum − + − − + Direct contact, indirect 8
Fungus Trichophyton sp. − + − − + Direct contact, indirect 8
*“Wild” refers to free-ranging wild orangutans who have never been in captivity or come into close human contact.
†
“Rehab” includes rehabilitation centers and zoos within the orangutan home range of Indonesia and Malaysia.
‡
“Zoo” includes zoos, laboratories, and private collections outside of Indonesia and Malaysia.
§
Letter of Concern raised against this study; see text.
¶
Antibodies against the following Leptospira serovars were found: australis, autumnalis, ballum, bratislava, grippotyphosa, hardjo, icterohaemorrhagiae, pyrogenes, saxkoebing, serjoe, szwajizak, wolffi.
HAV, Hepatitis A virus; HBV, hepatitis B virus; HTLV, human T-lymphotropic virus; SFV, simian foamy virus; SRV, simian D-type retrovirus; STLV, simian T-lymphotropic virus.
CHAPTER 80 Infectious Diseases of Orangutans in their Home Ranges and in Zoos
569
570 SE C T I O N 16 Great Apes
recognized. Infections in zoo-based orangutans have caused TABLE Bacteria Cultured from Air Sacculitis Cases in
fatal disease, and EMCV antigen or specific antibodies have 80.2 Orangutans (Pongo spp.)
been documented in zoos.13,14
The majority of our knowledge of infectious diseases of Organism Zoo* Rehab.†
orangutans stems from serologic tests for antibodies. Most Aerobacter cloacae — 1
of these tests are validated for humans but not NHPs. Even
Aeromonas hydrophilia 1 —
in validated tests, there is known to be a certain level of
cross reactivity with closely related48 or unrelated antigens,49 Alcaligenes fecalis 1 —
which may make accurate diagnosis challenging. A prime Bacterioides spp. 2 —
example of this was a study published on the serologic
Enterobacter sp. 2 4
evidence of African strains of Ebola virus in orangutans in
Indonesia,19 the implications of which could have had a Enterococcus 1 —
critical effect on release potential for orangutans in reha- Escherichia coli 20 2
bilitation centers. Although it is possible that orangutans
Flavobacterium odoratus 1 —
carry antibodies against Asian filoviruses such as Ebola
Reston virus, it is highly unlikely that they have been in Klebsiella oxytoca — 1
contact with African filoviruses. In addition, there were Klebsiella pneumoniae 12 3
numerous factually wrong statements in the paper (origin of
Micrococcus sp. — 1
samples, sample collection methods), as well as questionable
methodology, rendering the conclusions unfounded, which Morganella morganii — 1
resulted in a published letter of concern.50 Proteus spp. 7 6
Unlike African great apes, there does not seem to be an
Pseudomonas aeruginosa 17 3
orangutan-specific simian immunodeficiency virus (SIV) in
the home range. Antibodies against SIV have previously Pseudomonas spp. 2 6
been found by enzyme-linked immunosorbent assay in 2 Staphylococcus spp. 5 2
of 19 orangutans in North American zoos, but confirma-
Streptococcus spp. 9 2
tion tests (Western blot) were negative.51
Enteric parasites and protozoa (especially Strongyloides, Yersinia sp. — 2
hookworm, Trichostrongyles, Balantidium coli, and Ent- Gram-negative bacilli 1 3
amoeba spp.) have a high prevalence in captive orangutans,
*Zoo cases (n = 33) from Wells et al. (1990, n = 7) and Zimmermann
in zoos as well as in home-range countries. Balantidium et al. (2011, n = 26).
appear to thrive under stress, regardless of the orangutan’s †
Rehabilitant cases (n = 25) from Lawson et al. (2006, n = 11) and
location. Strongyloides were reported to be the leading cause Borneo Orangutan Survival Foundation unpublished data (n = 14).
of death of orangutans younger than 15 years in zoos.37
These enteric parasites have also been documented in wild
orangutans.4,33–35
It is not in the scope of this chapter to go into detail for Enteric bacteria are often cultured from infected air sacs
every pathogen reported in orangutans or the treatment; (Table 80.2), suggesting that inhalation of fecal contami-
for such an overview, we refer the reader to Chapter 83.52 nants may initiate infection.8,53,57 Infection risk from fecal
Instead, we will highlight some of the biggest differences aerosolization has been correlated with husbandry factors
in infectious disease between orangutans in zoos and their such as lower ventilation and smaller sleeping areas56 and
home range, or those of greater importance with regard to cage cleaning while the animals are present.53 Additional
zoonotic or release potential. causal factors could be chronic sinusitis57,60 or respiratory
irritants: severe smoke pollution in Indonesia and Malaysia
Air Sacculitis from annual forest fires is linked to respiratory problems in
human residents61 and increased incidence of air sacculitis
Respiratory disease is the primary infectious health issue in rehabilitant orangutans.56
affecting zoo orangutans and is important in captive popu- In zoos, adults represent the majority of published
lations in home-range countries. A significant proportion of cases,54,55 whereas the highest incidence in rehabilitation
these cases are air sacculitis, where purulent material collects centers is in juveniles (2–8 years).56,57 This may reflect
in the laryngeal air sacs,53 which is frequently diagnosed the relative population structure in zoos, biased toward
both in zoos8,54,55 and in rehabilitant populations.56,57 Air older animals, compared with rehabilitation centers with
sacculitis is not thought to be contagious57; therefore it a younger population. A male sex bias has been found in
is likely that husbandry factors in captivity contribute to air sacculitis cases in rehabilitant juvenile orangutans,56 as
the incidence of this condition.53 There are no published in other NHPs.62 In zoos, males are predisposed to respira-
reports of air sacculitis in wild orangutans, contrary to other tory infections, although not specifically air sacculitis.55
free-ranging NHPs.53,58,59 There is marked sexual dimorphism in the air sac in adult
CHAPTER 80 Infectious Diseases of Orangutans in their Home Ranges and in Zoos 571
orangutans; it is possible that sex differences in air sac been reported.66 There have been very few cases of confirmed
structure are present at a young age. human HBV in confiscated orangutans, but the zoonotic
risk is high. It is of the utmost importance to confirm the
Vector-Borne Diseases strain of hepadnavirus before release into the forest, where
wild orangutans are naïve to the human strain. Preventative
In the orangutan’s home range, vector-borne infections measures e.g., PPE are essential. People carrying a blood-
are abundant, and malaria is the most common infectious borne zoonotic pathogen such as HBV should refrain from
disease in rehabilitation centers. Classically, two Plasmo- situations where the primate may bite the worker, which
dium species have been described only in orangutans, minimizes risk of transmission and is required in human
P. pitheci and P. sylvaticum40; neither of these has been health care.67 Antiviral drug therapy that reduces viral load
associated with clinical disease. Most Plasmodium species minimizes this risk further.
are considered to have a sylvatic cycle, where wild NHPs
are often asymptomatic carriers. However, in rehabilitation Tuberculosis
centers, orangutans with clinical signs (high fever, lethargy)
are commonly diagnosed with “human” Plasmodium species Tuberculosis (TB), caused by infection with Mycobacterium
(e.g., P. falciparum, P. vivax) by microscopy and respond tuberculosis complex bacteria, can affect all mammalian
positively to treatment with artemisinin-based medica- species.68 In addition to acute respiratory disease, infection
tions (authors’ experience). The presence of the different can remain latent and undetected for many years, making it
Plasmodium species (including P. knowlesi) was confirmed difficult to control69 and resulting in preventative euthanasia
by polymerase chain reaction (PCR) (unpublished) and in most infected captive NHPs. Cases of TB in orangutans
published for another rehabilitation center,41 although have been reported from zoos and rehabilitation centers,
this was later disputed.39 In centers, high primate popu- but it has never been documented in wild orangutans.8,70 In
lation densities at ground level, combined with a higher home-range countries, TB is endemic in the human popu-
density of mosquitoes (Culicidae) at ground level than lation69; thus increased human contact increases the risk
in the canopy,63 lead to an increased risk of Plasmodium of naïve wild-born orangutans to exposure to pathogenic
infection. mycobacteria. Even where TB is not endemic, zoos must
Arboviruses are abundant in the home ranges and also undertake regular TB testing to prevent entry and spread
have a sylvatic cycle, with nonclinical infections of NHPs. through their collections.68
Antibodies against dengue and other arboviruses have been Traditionally, the tuberculin skin test (TST) is used,53
described in semicaptive and free-ranging orangutans.4 but orangutans seem to be more sensitive and often show
At the Borneo Orangutan Survival Foundation (BOSF) a nonspecific T-cell response.8,31 A comparative TST is
rehabilitation center at Nyaru Menteng, febrile, lethargic, recommended, using antigen from tuberculous and nontu-
thrombocytopenic juvenile orangutans commonly tested berculous mycobacteria, to account for cross reactivity.31,53,70
positive in a Dengue-specific immunoglobulin M rapid test Using highly concentrated bovine purified protein deriva-
(designed for human use). We collected samples from these tive (100,000 IU/mL) and standard avian purified protein
animals: all tested negative by reverse transcriptase PCR derivative (25,000 IU/mL) and reading the greatest response
and for virus isolation. Serologic tests were negative for at either 48 or 72 hours provides the most reliable results
NS1 antigen and IgM antibodies, whereas several samples in orangutans.70 Positive diagnosis of active TB by culture
tested positive for immunoglobulin G (unpublished). The or PCR of gastric or bronchoalveolar samples is definitive
cross reactivity of antibodies between the different flavivi- but has low sensitivity (approximately 60%–70% in human
ruses, as well as other pathogens, is well documented, war- cases).71 Trials have been done on several immunoassays as
ranting further research into natural flavivirus infections in adjunctive diagnostics for TB,32,68,72 although none appear
orangutans. to be conclusive for orangutans.73 Further research into
these tests is required.
Hepatitis B
Conclusion
Like most hepadnaviruses, hepatitis B virus (HBV) is fairly
species specific. However, discovery of recombinant HBV Although there is some overlap in the infectious diseases
of human and NHP origin has confirmed the potential for of orangutans in zoos, rehabilitation centers, or in the
cross-species transmission,64 which is a serious concern for wild, others are compounded by factors related to captivity
release programs and potentially for zoos. The prevalence of and geography. The susceptibility of great apes to human
human HBV infection is relatively high in orangutan home pathogens—and vice versa—must be considered and pre-
ranges, whereas vaccination has limited the prevalence in ventative measures taken. Accurate detection of infectious
other countries. Orangutans have genetically very similar disease in orangutans is vital, both for conserving the health
HBVs, which appear to be nonpathogenic, but antibodies of zoo populations and to minimize the risk of “human”
cross-react with human HBVs.65 In confiscated orangutans pathogens being introduced to wildlife populations when
in the home range, a prevalence of up to 59% has previously rehabilitated orangutans are released to the wild. A good
572 SE C T I O N 16 Great Apes
understanding of the accuracy and limitations of the avail- 20. Murphy HW, Miller M, Ramer J, et al: Implications of simian
able diagnostic tests is key to achieving this. retroviruses for captive primate population management and
the occupational safety of primate handlers, J Zoo Wildl Med
37(3):219–233, 2006.
Acknowledgment 21. Pettersson JH-O, Aspán A, Krützen M, et al: Staphylococcal and
Streptococcal zoonoses in wild Bornean and Sumatran orang-
Borneo Orangutan Survival Foundation (BOSF) utans. 10th International Meeting on Microbial Epidemiological
Markers, Paris, October 2–5, 2013.
References 22. Chai N, Hazan T, Wedlarski R, et al: Treatment of a retroperito-
neal abscess by omentalization in an orangutan (Pongo pygmaeus
1. Ancrenaz M, Gumal M, Marshall AJ, et al: Pongo pygmaeus. pygmaeus), J Zoo Wildl Med 40(2):350–353, 2009.
The IUCN Red List of Threatened Species 2016. Report No.: e. 23. Brack M: The Department of Pathology at the German Primate
T17975A17966347. Center from 1973 to 1999, Primate Biol 2(1):81, 2015.
2. Singleton I, Wich SA, Nowak M, et al: Pongo abelli. (errata 24. Meier JE, Sanborn W: A preliminary report on the manage-
version published in 2016) The IUCN Red List of Threatened ment and treatment of salmonellosis with trimethoprim-
Species 2016. Report No.: e. T39780A10266609. sulfamethoxazole in an exotic animal nursery, J Zoo Anim Med
3. Buitendijk H, Fagrouch Z, Niphuis H, et al: Retrospective serol- 13(1):26–29, 1982.
ogy study of respiratory virus infections in captive great apes, 25. Rahmi E, Agustina D, Jamin F: Isolation and identification of
Viruses 6(3):1442–1453, 2014. Genus Salmonella and Shigella from Sumatran orangutan (Pongo
4. Kilbourn AM, Karesh WB, Wolfe ND, et al: Health evaluation abelii) feces in Orangutan Reintroduction Center, Jantho, Jurnal
of free-ranging and semi-captive orangutans (Pongo pygmaeus Medika Veterinaria 8(1):2014.
pygmaeus) in Sabah, Malaysia, J Wildl Dis 39(1):73–83, 2003. 26. Yeager C, Andayani N, Eudey A, et al: Orangutan Action Plan.
5. Groenewoud MJ, Fagrouch Z, van Gessel S, et al: Character- 1998.
ization of novel polyomaviruses from Bornean and Sumatran 27. Pazzaglia G, Widjaja S, Soebekti D, et al: Persistent, recurring
orangutans, J Gen Virol 91(3):653–658, 2010. diarrhea in a colony of orangutans (Pongo pygmaeus) caused by
6. Kalter SS, Heberling RL, Cooke AW, et al: Viral infections of multiple strains of Campylobacter spp, Acta Trop 57(1):1–10,
nonhuman primates, Comp Med 47(5):461–467, 1997. 1994.
7. Warren KS: Orangutan conservation: epidemiological aspects of 28. Smith N, Damit M: Fatal bronchopneumonia in a young
health management and population genetics. Murdoch Univer- orangutan caused by Pseudomonas pseudomallei, Vet Rec 110(11):
sity, 2001. 251, 1982.
8. Wells SK, Sargeant E, Andrews ME, et al: Medical management 29. Ketz-Riley CJ, Kennedy GA, Carpenter JW, et al: Tularemia type
of the orangutan. New Orleans, LA, The Audubon Institute, a in captive Bornean orangutans (Pongo pygmaeus pygmaeus), J
1990. Zoo Wildl Med 40(2):257–262, 2009.
9. Rasheed S, Rongey RW, Bruszweski J, et al: Establishment of a 30. Schrenzel MD, Witte CL, Bahl J, et al: Genetic characterization
cell line with associated Epstein-Barr-like virus from a leukemic and epidemiology of helicobacters in non-domestic animals,
orangutan, Science 198(4315):407–409, 1977. Helicobacter 15(2):126–142, 2010.
10. Sakulwira K, Theamboonlers A, Oraveerakul K, et al: Orangutan 31. Calle PP, Thoen CO, Roskop ML: Tuberculin skin test responses,
herpesvirus, J Med Primatol 33(1):25–29, 2004. mycobacteriologic examinations of gastric lavage, and serum
11. Arita I, Henderson DA: Smallpox and monkeypox in non-human enzyme-linked immunosorbent assays in orangutans (Pongo
primates, Bull World Health Organ 39(2):277, 1968. pygmaeus), J Zoo Wildl Med 20:307–314, 1989.
12. Miyagi J, Tsuhako K, Kinjo T, et al: Coxsackievirus B4 myocar- 32. Kilbourn AM, Godfrey HP, Cook RA, et al: Serum antigen 85
ditis in an orangutan, Vet Pathol 36(5):452–456, 1999. levels in adjunct testing for active mycobacterial infections in
13. Canelli E, Luppi A, Lavazza A, et al: Encephalomyocarditis virus orangutans, J Wildl Dis 37(1):65–71, 2001.
infection in an Italian zoo, Virol J 7(1):64, 2010. 33. Foitová I, Huffman MA, Wisnu N, et al: Parasites and their
14. Citino SB, Homer BL, Gaskin JM, et al: Fatal encephalomyo- impacts on orangutan health. In Wich SA, Utami Atmoko SS,
carditis virus infection in a Sumatran orangutan (Pongo pygmaeus Mitra Setia T, et al, editors: Orangutans: geographic variation in
abelii), J Zoo Anim Med 214–218, 1988. behavioral ecology and conservation, Oxford, UK, 2009, Oxford
15. Kalter SS, Heberling RL: Comparative virology of primates, University Press, pp 157–167.
Bacteriol Rev 35(3):310, 1971. 34. Labes EM, Hegglin D, Grimm F, et al: Intestinal parasites of
16. Harrison VR, Marshall JD, Guilloud NB: The presence of endangered orangutans (Pongo pygmaeus) in Central and East
antibody to Chikungunya and other serologically related viruses Kalimantan, Borneo, Indonesia, Parasitology 137(1):123, 2009.
in the sera of sub-human primate imports to the United States, 35. Mul IF, Paembonan W, Singleton I, et al: Intestinal parasites of
J Immunol 98(5):979–981, 1967. free-ranging, semicaptive, and captive Pongo abelii in Sumatra,
17. Fahroni A, Tandang M, Patispathika FH, et al: First report of Indonesia, Int J Primatol 28(2):407–420, 2007.
rabies in an orangutan. Borneo Orangutan Survival Foundation, 36. Canfield PJ, Vogelnest L, Cunningham ML, et al: Amoebic
2014. meningoencephalitis caused by Balamuthia mandrillaris in an
18. Chen C-C, Pei K-C, Yang C-M, et al: A possible case of orangutan, Aust Vet J 75(2):97–100, 1997.
hantavirus infection in a Borneo orangutan and its conservation 37. Lung N, Smith J: Orangutan SSP Veterinary Advisors Report.
implication, J Med Primatol 40(1):2–5, 2011. 2013.
19. Nidom CA, Nakayama E, Nidom RV, et al: Serological evidence 38. Hanley CS, Simmons HA, Wallace RS, et al: Visceral and
of Ebola virus infection in Indonesian orangutans, PLoS ONE presumptive neural baylisascariasis in an orangutan (Pongo
7(7):e40740, 2012. pygmaeus), J Zoo Wildl Med 37(4):553–557, 2006.
CHAPTER 80 Infectious Diseases of Orangutans in their Home Ranges and in Zoos 573
39. Singh B, Divis PCS: Orangutans not infected with Plasmodium 59. Köndgen S, Leider M, Lankester F, et al: Pasteurella multocida
vivax or P. cynomolgi, Indonesia, Emerg Infect Dis 15(10):1657, involved in respiratory disease of wild chimpanzees, PLoS ONE
2009. 6(9):e24236, 2011.
40. Peters W, Garnham PCC, Killick-Kendrick R, et al: Malaria of 60. Steinmetz HW, Zimmermann N: Computed tomography for the
the orangutan (Pongo pygmaeus) in Borneo, Philos Trans R Soc diagnosis of sinusitis and air sacculitis in orangutans. In Fowler
Lond B Biol Sci 275(941):439–482, 1976. ME, Miller RE, editors: Zoo and wild animal medicine: current
41. Reid MJ, Ursic R, Cooper D, et al: Transmission of human therapy, ed 7, Philadelphia, 2012, Saunders.
and macaque Plasmodium spp. to ex-captive orangutans in 61. Koplitz SN, Mickley LJ, Marlier ME, et al: Public health impacts
Kalimantan, Indonesia, Emerg Infect Dis 12(12):1902, 2006. of the severe haze in Equatorial Asia in September–October
42. Elder M: 2011 International Studbook of the Orangutan (Pongo 2015: demonstration of a new framework for informing fire
pygmaeus, Pongo abelii), 2012, p 261. management strategies to reduce downwind smoke exposure,
43. Wolfe ND, Kilbourn AM, Karesh WB, et al: Sylvatic transmis- Environ Res Lett 11(9):094023, 2016.
sion of arboviruses among Bornean orangutans, Am J Trop Med 62. Kumar S, Fox B, Owston M, et al: Pathology of spontaneous
Hyg 64(5):310–316, 2001. air sacculitis in 37 baboons and seven chimpanzees and a brief
44. Kik MJ, Bos JH, Groen J, et al: Herpes simplex infection in a review of the literature, J Med Primatol 41(4):266–277, 2012.
juvenile orangutan (Pongo pygmaeus pygmaeus), J Zoo Wildl Med 63. Brant HL, Ewers RM, Vythilingam I, et al: Vertical stratifica-
36(1):131–134, 2005. tion of adult mosquitoes (Diptera: Culicidae) within a tropical
45. Borneo Orangutan Survival Foundation: Unpublished data. rainforest in Sabah, Malaysia, Malar J 15(1):370, 2016.
2017. 64. Bonvicino CR, Moreira MA, Soares MA: Hepatitis B virus
46. Calattini S, Betsem EB, Froment A, et al: Simian foamy virus lineages in mammalian hosts: potential for bidirectional cross-
transmission from apes to humans, rural Cameroon, Emerg Infect species transmission, World J Gastroenterol 20(24):7665, 2014.
Dis 13(9):1314, 2007. 65. Warren KS, Heeney JL, Swan RA, et al: A new group of hepad-
47. Jones-Engel L, Engel GA, Schillaci MA, et al: Primate-to-human naviruses naturally infecting orangutans (Pongo pygmaeus), J Virol
retroviral transmission in Asia, Emerg Infect Dis 11(7):1028, 73(9):7860–7865, 1999.
2005. 66. Warren KS, Niphuis H, Verschoor EJ, et al: Seroprevalence of
48. MacNeil A, Reed Z, Rollin PE: Serologic cross-reactivity of specific viral infections in confiscated orangutans (Pongo pyg-
human IgM and IgG antibodies to five species of Ebola virus, maeus), J Med Primatol 27(1):33–37, 1998.
PLoS Negl Trop Dis 5(6):e1175, 2011. 67. Henderson DK, Dembry L, Fishman NO, et al: SHEA guideline
49. Bhatti AB, Ali F, Satti SA: Cross-reactivity of rapid Salmonella for management of healthcare workers who are infected with
Typhi IgM immunoassay in dengue fever without co-existing hepatitis B virus, hepatitis C virus, and/or human immunode-
infection, Cureus 7(12):e396, 2015. ficiency virus, Infect Control Hosp Epidemiol 31(03):203–232,
50. The PLoS ONE Editors: Expression of concern: serological 2010.
evidence of ebola virus infection in indonesian orangutans, PLoS 68. Miller MA, Lyashchenko KP: Mycobacterial infections in other
ONE 8(3):e60289, 2013. zoo animals. In Mukundan H, Chambers M, Waters R, et al,
51. Lowenstine LJ, Pedersen NC, Higgins J, et al: Seroepidemiologic editors: Tuberculosis, leprosy and mycobacterial diseases of man and
survey of captive old-world primates for antibodies to human animals: the many hosts of mycobacteria, Boston, MA, 2015, CAB
and simian retroviruses, and isolation of a lentivirus from International, pp 277–295.
sooty mangabeys (Cercocebus atys), Int J Cancer 38(4):563–574, 69. World Health Organization: Global Tuberculosis Report 2012.
1986. WHO, 2012.
52. Murphy HW: Great apes. In Miller RE, Fowler ME, editors: 70. Dench R, Sulistyo F, Fahroni A, et al: Evaluation of diagnostic
Fowler’s zoo and wild animal medicine, 2015, Elsevier Health accuracy of the comparative tuberculin skin test in reha-
Sciences, pp 336–354. bilitant Bornean orangutans (Pongo pygmaeus), J Zoo Wildl Med
53. McManamon R: Veterinary medical management. In Sodaro 46(4):833–842, 2015.
C, editor: Orangutan species survival plan husbandry manual, 71. Negi SS, Khan SFB, Gupta S, et al: Comparison of the conven-
Chicago, IL, 2007, Chicago Zoological Park, pp 221–291. tional diagnostic modalities, bactec culture and polymerase chain
54. Herrin KA, Spelman LH, Wack R: Surgical air sac resection reaction test for diagnosis of tuberculosis, Indian J Med Microbiol
as a treatment for chronic air sacculitis in Great Apes. Pro- 23(1):29–33, 2005.
ceedings of the American Association of Zoo Veterinarians, 72. Lyashchenko KP, Greenwald R, Esfandiari J, et al: PrimaTB
2002. STAT-PAK assay, a novel, rapid lateral-flow test for tuberculosis
55. Zimmermann N, Pirovino M, Zingg R, et al: Upper respira- in nonhuman primates, Clin Vaccine Immunol 14(9):1158–1164,
tory tract disease in captive orangutans (Pongo sp.): prevalence 2007.
in 20 European zoos and predisposing factors, J Med Primatol 73. Unwin S, Colin C, Nente C, et al: Stat-pak(R) blood assays in M.
40(6):365–375, 2011. tuberculosis complex surveillance testing in populations of chim-
56. Dench R, Tandang M, Sulistyo F, et al: Risk factors for air panzees (Pan sp) and orangutans (Pongo sp) in range countries.
sacculitis in rehabilitant Bornean orangutans (Pongo Pygmaeus). Proceedings of the EAZWV/IZW International Conference on
Borneo Orangutan Survival Foundation, 2014. Diseases of Zoo and Wild Animals. Bussolengo, Italy: European
57. Lawson B, Garriga R, Galdikas BM: Air sacculitis in fourteen Association of Zoo and Wildlife Veterinarians/Leibniz Institute
juvenile southern Bornean orangutans (Pongo pygmaeus wurmbii), for Zoo and Wildlife Research, 2012, p 109.
J Med Primatol 35(3):149–154, 2006. 74. Roy S, Vandenberghe LH, Kryazhimskiy S, et al: Isolation
58. Hastings BE: The veterinary management of a laryngeal air sac and characterization of adenoviruses persistently shed from
infection in a free-ranging mountain gorilla, J Med Primatol the gastrointestinal tract of non-human primates, PLoS Pathog
20(7):361, 1991. 5(7):e1000503, 2009.
81
Medical Aspects of
Chimpanzee Rehabilitation
and Sanctuary Medicine
JOCELYN BEZNER
574
CHAPTER 81 Medical Aspects of Chimpanzee Rehabilitation and Sanctuary Medicine 575
Much of the following primate medicine information is consistently anxious during the process may benefit from
from a private sanctuary that cares for 248 chimpanzees, a partnership with a conspecific with excellent social skills.
many of whom came from a financially failing biomedical Once socially bonded, triadic and small group formations
laboratory. Others arrived from roadside zoos, entertain- commence. Every relationship has its own time period and
ment venues, and the pet trade. The 150-acre sanctuary is deemed satisfactory if they fulfill behavioral benchmarks
houses 237 chimpanzees divided into 12 families made up such as grooming, playing, and other affiliative behavior.
of both males and females of varying ages. Each group has However, the personality of the participants must be taken
an indoor living space with six interconnected bedrooms into consideration; otherwise, strictly defined criteria of
attached to a 3- to 4-acre grass-covered island with trees, a satisfactory pairing may impede an animal’s successful
hills, climbing structures, and platforms. Eleven animals entry into a social group. Finally, chimpanzees who do
deemed psychologically or medically unfit for large social not exhibit species-appropriate behavior during initial
groups live with fewer chimps in a fission-fusion pattern in introductions may learn the nuances of being a chimp
a building with separate large outdoor yards. Two of these during repetitive positive interactions that may lead to sub-
chimpanzees are singly housed for psychological reasons sequent successful results. After each family member meets
but have close visual, auditory, and safe tactile contact with individually, different configurations of small subgroups
neighboring chimps. Attempts are ongoing to acclimate are carefully combined to identify relationships that may
them with conspecifics. be detrimental and need resolution prior to unity of the
entire group.8,11
Housing and Introductions The behavioral and veterinary departments work col-
laboratively if psychopharmacologic treatment is deemed
Captive chimpanzees have unique, complex personalities, necessary for the health and well-being of particular
diverse backgrounds, and abnormal rearing histories when animals. Definitive guidelines for behavioral drugs in
raised for research, entertainment, or as pets, making it chimpanzees are lacking, and therapy is based on human
a challenge to fit them into cohesive hierarchal groups.4 recommendations.14,15 Short-term use of anxiolytic ben-
Human-rearing of chimpanzees and other nonhuman pri- zodiazepines has a rapid effect and good tolerability and
mates has undergone a significant shift in zoos and is now helps chimpanzees that display inappropriate fear, anxiety,
done only when deemed necessary and emphasizes an early agitation, or mild forms of self-injury during introduc-
return to conspecifics.5,6 Social housing, defined by GFAS, tions or in general. There is wide variability in dosing
requires that the chimpanzees be in a compatible group between animals, and response to therapy and lack of
without physical or psychological stress and that there is side effects dictate both the dose and treatment length.
adequate space to allow for more natural fission-fusion Diazepam at 5–30 mg administered once (SID) to twice
affiliations.7 Chimpanzee introductions require knowledge (BID) daily appears to work better than lorazepam at
of the individual’s personality, a comprehensive understand- 0.5–3 mg SID to BID. For introductions, diazepam may
ing of chimpanzee behavior, and a building design that be stopped and started as needed. If used consistently, slow
allows for quick and safe separation of chimpanzees if the weaning is recommended to prevent acute withdrawal
introduction does not go as anticipated. Resocialization of symptoms.15
socially deprived chimpanzees has been successful at multiple Aggression, though fairly common in chimpanzee societ-
sanctuaries, zoos, and laboratories, and various techniques ies, may hamper an individual’s opportunity to live in a
have been described elsewhere in the literature.8–11 As large, complex community in captivity. Every effort should
noted, the number of chimpanzees arriving at sanctuaries is be made to help facilitate this process. Psychopharmacologic
increasing due to the recent ban on their use in biomedical treatment from available veterinary and human literature is
research. Many of them do not have the species’ typical selected based on efficacy, taste, acceptance, and side effects.
communication or social skills that wild chimpanzees learn Drug selection must take into account the therapeutic index
from an extensive and long maternal childhood and sup- and hematological/biochemical effects of the medicine.
portive complex social communities.12,13 Groups ranging in Done judiciously, this will reduce the frequency and stress
numbers from 12–27 chimpanzees are formed by dyadic of venipuncture by positive reinforcement training or seda-
introductions of all individuals, and mild aggression or tion. Blood drug levels, common in human medicine, may
tension often resolves spontaneously without intervention. be more difficult to obtain in primates.
If aggression is deemed moderate to severe from the outset or The central sympatholytic effects of the alpha-2
escalates, the individuals are separated for a period of 5–15 adrenergic receptor agonist, clonidine, is used to treat dif-
minutes. During this “time out” the chimps are maintained ferent forms of aggression in people. It appears to have
in close proximity separated by mesh. Reintroduction com- similar effects in chimpanzees. An oral starting dose of
mences once the arousal dissipates and frequently results in 0.05–0.1 mg daily can be titrated up to 0.2 mg SID to
affiliative interaction. If avoidance or aggressive behavior BID depending on response and side effects. Guanfacine,
does not resolve, the introduction is discontinued for a a sister compound, may also be used but is less sedating.18
period of time ranging from a day to months while other A demonstrated change in temperament manifested by less
dyadic relationships commence. Chimpanzees appearing piloerection, aggressive displays, and violence is considered
576 SE C T I O N 16 Great Apes
a positive response. The drug may be continued for months, may not elicit a reaction, but a response to a light spray of
but slow weaning must be done to avoid rebound water on the face will determine if supplemental drugs are
hypertension.17 necessary.
Risperidone is an atypical antipsychotic used for human Oral transmucosal medetomidine, 0.1 mg/kg, has been
psychiatric conditions including aggression. Controlled administered in marshmallow crème or applesauce in
data is lacking for the use of this drug in chimpanzees chimpanzees in combination with tiletamine-zolazepam.26
but was tried on seven aggressive males that failed to Attempts to use oral medetomidine prior to IM injection of
assimilate into any large group after multiple attempts and an induction agent was not successful due to the inconsis-
configurations. The primates received oral risperidone, 0.02 tency of the sedation, variable time to effect, and cyanotic
mg/kg–0.07 mg/kg divided BID. All the chimpanzees were mucous membranes.
successfully integrated when treated with risperidone. The Once anesthesia is induced, supplementation with
chimpanzees showed no side effects and were subsequently ketamine at a dose of 2–5 mg/kg IM or gas anesthesia
weaned at 6 months to one year, though one chimp was with isoflurane is selected based on the length of the
treated intermittently for 2 years. They have all remained procedure and depth of anesthesia required.23 With the
in complex social environments. chimpanzee in dorsal recumbency, the vocal folds are easily
Use of haloperidol, a conventional antipsychotic, was visualized by placing a lighted laryngoscope with a long
discontinued because oral doses of 5–10 mg/day produced curved blade into the vallecula and gently pulling up. No
bradykinesia in five male chimpanzees. Haloperidol has laryngeal spasms or inability to intubate occurred in over
been used successfully in one female chimpanzee for 1000 intubations using a dry 6–8 cuffed endotracheal tube
generalized anxiety and intermittent self-mutilation at with a stylet. Once the tube is secured, the table is tilted
2–4 mg/day. to remove the pressure from the abdominal organs on
the diaphragm. A surgical monitor measuring peripheral
partial oxygen pressure (SpO2), heart rate, end tidal carbon
Physical Examinations and Anesthesia dioxide, electrocardiogram, and indirect blood pressure is
in Sanctuaries recommended. The use of intravenous fluids, preemptive
antibiotics, analgesics, local blocks, and temperature main-
Clinical signs, age, history, risk-to-benefit ratio of anes- tenance is determined by the patient and the procedure.
thesia, and familiarity with the chimpanzee determine In the event of an anesthesia complication, medetomidine
the frequency of physical examinations and the anesthetic can be quickly reversed by dividing the total dose of atipa-
protocol. Chimpanzees from laboratories are suspicious mezole (5 mg per 1 mg medetomidine) and administering
when separated from their companions, and sedation is one-fourth of the dose intravenously and the remainder
best achieved in the morning after a 12–16-hour fast with IM.23 Supplemental anesthesia should be immediately
no water restrictions. The chimpanzee can receive an oral available to circumvent early arousal from reversal.23
dose of diazepam (10–40 mg) in juice 1–2 hours prior to Chimpanzees recovering from noncomplicated anesthesia
anesthesia to decrease anxiety. Operant conditioning for are placed in lateral recumbency with the upper arm posi-
acceptance of injections is useful, though time-consuming tioned behind the back, and the head placed close to an
for a large population of chimpanzees.22 Drugs and doses area where extubating and suctioning of oral secretions is
for great apes have been previously reported.22–24 A posi- safely achieved behind a protective barrier. If atipamezole
tive relationship between the veterinarian and the primate is used to reverse the medetomidine, waiting a minimum
may facilitate hand injections using a greeting or a small of 45 minutes after the last dose of ketamine prevents
sip of juice as a distraction. Healthy animals may first undesirable emergence reactions from the dissociative
be given the intramuscular (IM) sedative medetomidine anesthetic and results in a smooth recovery within 7–15
(ZooPharm, Laramie, Wyoming, USA; 0.02–0.05 mg/ minutes. Not reversing the alpha-2 adrenergic agonist also
kg, 10 mg/mL) in a 1 cc Luer Lock syringe and 23–25 g prevents emergence delirium and may be advantageous
⅝ inch needle held out of view or hidden inside a loose for postsurgical or painful cases. Tiletamine-zolazepam
latex glove. The chimps are injected into any body area results in prolonged recoveries alone or after reversal of the
closest to the mesh directly or through the latex glove. medetomidine.23
Quiet reassurance from the veterinarians and/or care staff Medetomidine is not recommended for suspected or
keeps the chimp close until complete recumbent sedation confirmed cardiac disease, sick, elderly, or other high-risk
occurs within 3–15 minutes. An immobilization dose for patients due to potential changes in cardiovascular func-
ketamine or tiletamine-zolazepam is then administered tion.24 Protocols using ketamine, ketamine/midazolam,
IM by hand or pole syringe with no reaction from the or tiletamine–zolazepam are thoroughly outlined else-
chimpanzee.22 Chimpanzees from research backgrounds where.22–24 Reconstitution of tiletamine-zolazepam to
generally require the higher dose range. Respiratory rate, 200 mg/mL lowers the volume allowing for more successful
the color of the mucous membranes, and heart rate are hand injections. All high-risk patients should be intubated
monitored during the induction period for an additional and receive oxygen alone or gas anesthesia and be monitored
10 minutes. Shaking or otherwise stimulating the chimp with a surgical monitor.
CHAPTER 81 Medical Aspects of Chimpanzee Rehabilitation and Sanctuary Medicine 577
Sanctuary Preventative Healthcare flat on the ends of the VD for coagulation. This technique
resulted in 86% regrowth and seven unwanted births. The
Preventative healthcare programs at sanctuaries follow surgery was modified to cauterize only the epithelium of
similar guidelines as those at zoos.25,27 Exceptions may the lumen by introducing a nonheated fine wire electrode
include pretransport exams, length of quarantine time, (Jorgensen JO470DT5) into each end of the VD once a
immunoprophylaxis, and reproductive methods. Animals portion of it was removed. The electrocautery unit is turned
being transferred to sanctuaries may have minimal to no to coagulation as the tip is withdrawn. Single portable
previous veterinary care. Lack of veterinarians, a seizure battery thermocautery units may be used by pinching the
situation, or the volatile emotions of those relinquishing tip with a sterile hemostat to facilitate introduction into
the primate may necessitate doing the physical examina- the small lumen. After intraluminal cautery, recanalization
tion, laboratory specimen collections, and intradermal is further prevented by fascial interposition of the VD
tuberculin testing on the day of transport. A veterinarian ends, followed by subcutaneous and subcuticular suture
should accompany the animal during transport to mitigate closure.30 The chimpanzees do not pick at the surgical site
complications that could occur with an incomplete medical if chromic gut (3-0) is used for subcuticular sutures, but
background. Quarantine is essential to prevent the risk polydioxanone (PDS) caused significant postoperative self-
of disease exposure to others, but the length is based on grooming. Exploratory surgery 6 months to 2 years later
the source of the animals, test results, health status, and confirmed no regrowth following this technique in novel
development of clinical symptoms. Similarly, all employees animals and those that had a previous vasectomy. However,
are tuberculin tested yearly and educated on both zoonotic since histopathologic confirmation of VD was not done at
and anthropozoonotic diseases. the time of the initial surgery, it is unclear how much the
Routine parasite screening includes bacterial fecal cul- vasectomy technique influenced the outcome.
tures and fecal examinations by direct wet mounts and
centrifugal floatation with a commercial solution. Regular
group samplings of freshly defecated stools and individual Sanctuary Case Reports
testing are recommended as a screening tool and for those Ocular Herpes
with gastrointestinal symptoms. Plain fecal samples and feces
in 2% formalin are intermittently submitted to a reference Alphaherpesviruses in great apes have been identified as
laboratory for confirmation. Fecal cultures on all animals at human simplex virus groups 1 and 2 (HSV1 and HSV2).31
the sanctuary have been consistently negative for Shigella, Several young chimps developed acute white oral/pharyngeal
Salmonella, Yersinia, Campylobacter, and Escherichia coli. ulcers that resolved without treatment. Viral culture and
Examination and sampling of the large and small intestines genome sequencing on the virus isolated from the oral
at necropsies may provide information on parasite burden. lesions identified a new herpes virus, ChHV. This new
Balantidium coli, Troglodytella sp., Entamoeba, Enterobius, virus is antigenically similar but not a variant of the human
Giardia, and Strongyloides sp. have all been identified in simplex viruses, and further studies determined that ChHV
captive great apes.25,27,28 Proactive diagnostic and control and HSV2 are more closely related to each other than either
strategies are mandatory as self-medication for parasites is is to HSV1.32,33 Serum was tested on 42 chimpanzees and
severely limited in captivity.29 Control of internal parasites 35% of these chimps had antibodies to the ChHV virus.
has been successful with the use of fenbendazole (10 mg/kg The most common clinical presentation of herpes at
PO) and ivermectin (200 mcg/kg PO [repeated again in 2 the sanctuary is acute ocular pain with or without corneal
weeks]) alternated quarterly. Balantidium sp., an intestinal ulceration. The animals develop severe photophobia and are
protozoa, does not generally cause intestinal symptoms. If unable to keep their eyes open. Corneal scrapings on mul-
no other causes of diarrhea can be identified and there is no tiple animals for viral isolation were negative, yet a dramatic
response to dietary and nonpharmacologic therapy, treat- response occurred with antiviral medications. Valacyclovir,
ment of Balantidium with doxycycline, 100 mg BID, for 1 g per os (PO) twice daily or acyclovir 400 mg PO every
10 days may reduce the number of cysts and trophozoites 6 hours significantly shortened the clinical signs from weeks
and resolve the symptoms. Metronidazole is highly effective, to days. Having astute and well-informed caregivers able to
but acceptance is difficult even with compounded formulas. recognize early warning signs shortened the course of the
disease to 24 hours. The suspected mechanism is reactiva-
Reproductive Management tion of the latent virus from the ophthalmic branch of the
trigeminal nerve.33
Sanctuaries do not breed animals and there are different
methods of prevention (see Chapter 22). Nonreversible Demyelinating Disease
male sterilization is the preferred method for permanent
contraception at many primate sanctuaries. Vasectomies are An 11-year-old male chimpanzee previously used in
performed through a prescrotal incision in chimpanzees. The hepatitis C biomedical research developed acute bilateral
vas deferens (VD) is isolated, and up to 2–3 cm is removed. weakness of both legs and began to drool. The chim-
Previous methodology involved placing the cautery unit panzee was anesthetized with tiletamine-zolazepam. The
578 SE C T I O N 16 Great Apes
examination was unremarkable, but the complete blood success. Fluoxetine starting at 10 mg/day is increased to
count (CBC) showed leukocytosis 18,200 µL, with 20 mg after 1 week. If there is no therapeutic response in
neutrophilia (12,740 µL) and monocytosis (1820 µL).34 2–3 months, titration to the maximum dose of 60–80 mg/
The chimpanzee improved but 6 months later developed day is attempted to determine effectiveness. Selective
intermittent drooling, screaming, progressive leg weakness, serotonin norepinephrine reuptake inhibitors (SNRIs) are
and an obtunded level of consciousness. The blood chem- a newer class of antidepressants showing promise for treat-
istry, CBC, bile acids, fungal serology tests, and urinalysis ment of multiple disorders in humans.15 SNRIs should be
were normal. Viral results were negative for antibodies to used judiciously due to their known withdrawal symptoms
human immunodeficiency virus, simian immunodeficiency even with one to two missed doses and lack of compliance.
virus, HSV1 and 2, and the antigen for hepatitis C by However, one chimpanzee who severely self-mutilated his
polymerase chain reaction. The chimpanzee had antibodies head for 7 years was responsive to the off-label use of the
to cytomegalovirus and the Epstein-Barr virus. Magnetic SNRI milnacipran with a dose of 50 mg BID, and attempts
resonance imaging without and with gadolinium contrast- to wean the drug caused a resurgence of self-injury.15,40
enhanced T1-weighted images showed abnormal signaling Intensive behavioral and husbandry management and
in the white matter of both frontal lobes extending into the multiple drug trials were less effective.40
genu of the corpus callosum and descending white matter Neuroleptics (antipsychotics) block dopamine D2 recep-
tracts. The chimpanzee also had elevated very-long-chain tors in the brain.14–16 The typical antipsychotic, haloperidol,
fatty acids (VLCFA) consistent with a defect in peroxisomal produced extrapyramidal signs of dystonia and akathisia
fatty acid oxidation. He was euthanized 8 months later due in all male chimpanzees within hours to days after admin-
to deteriorating symptoms, and postmortem histopathology istration of 5–20 mg/day. The second-generation atypical
confirmed a diagnosis of demyelination. neuroleptics have different receptor affinities and produced
no side effects at doses used.15,16 Risperidone, 1.0–6.0 mg
Behavioral Issues divided BID, appears to be successful for generalized anxiety
as well as for aggression as discussed under Housing and
Nonhuman primates in zoos, sanctuaries, and laboratories Introductions earlier in the chapter.
often display behavioral abnormalities such as anxiety, Evidence-based science on psychopharmacology is
social isolation, stereotypies, displacement disorders, self- essential as chimpanzees move from laboratories to sanctu-
mutilation, and other aberrances. Facilities have different aries, yet there is a lack of data. Not all behaviors warrant
rates of these behaviors, and research is ongoing to improve treatment or can be helped with medication. One male
captive care by understanding the causes of these devia- chimpanzee severely mutilated his arm for 17 years in the
tions from the norm.35–37 Assessing the psychological health laboratory. Multiple sequential pharmacologic trials had
of animals is challenging, and some experts suggest the minimal effects. We will never know if better husbandry,
use of the human Diagnostic and Statistical Manual of behavioral enrichment, relationships with conspecifics,
Mental Disorders, 4th Edition (DSM-IV) to categorize the human bonding, surgeries, the 72-hour negative pressure
psychological disorders of great apes, but others have made a wound treatment with constant rate infusion (CRI) of
valid argument against it.14,35,36 The first step in dealing with ketamine, or the freedom to roam a 3-acre island helped
acute or chronic behavior patterns that interfere with the him recover, but he has stopped. The key point is to keep
quality of the sanctuary animal’s life is to search for and treat trying multiple modalities.
medical issues that may be contributory.15 Next, a compre-
hensive, individualized behavioral plan is designed based End of Life and Euthanasia
on symptoms, frequency, and severity. The use and timing
of pharmacologic therapy depends on multiple factors. The As the population of captive chimpanzees age, geriatric
genetic physiologic similarities between chimpanzees and medical issues develop that must be addressed to ensure
humans and the lack of rigorous systemic research support optimal veterinary care.40–43 Adding to the discussion is
extrapolation of treatment from human literature.14,15,37 All the diversity of the rearing history, nutrition, research, and
drugs selected in consultation with psychiatrists, behavior- previous veterinary care that may influence the type and
ists, and literature review must be considered empirical progression of disease in older primates.34,41,42 Ideally, the
at this time. Therefore the veterinarian must closely geriatric animal should stay with their social companions
monitor for signs of efficacy, safety, and tolerability. Well- forever, provided there are husbandry, structural, and vet-
planned and scheduled attempts at weaning help to justify erinary medicine modifications. Ramps, ladders, platforms,
a drug’s use. and straps facilitate mobility. Temporary separation from
Longer treatment options for anxiety, social isolation, the group for feeding or resting ensures less stress, proper
aggression, or compulsive disorders are the selective sero- nutrition, and time for the administration of medication
tonin reuptake inhibitors (SSRIs).14–16,37,38 Consulting psy- and evaluation by the veterinarian. Natural or planned
chiatrists recommended fluoxetine due to its proven safety formation of a smaller subgroup that facilitates daily
and the ability to discontinue the drug rapidly. Selection of access to the outside, opportunities for companionship,
different SSRIs for specific disorders may improve treatment and a decreased risk of trauma is beneficial. Just as every
CHAPTER 81 Medical Aspects of Chimpanzee Rehabilitation and Sanctuary Medicine 579
sanctuary animal has a specific medical and behavioral plan, 8. Seres M, Aurelli F, De Wall F: Successful formation of a large
end-of-life decisions are also individualized. Nutrition, chimpanzee group out of two preexisting subgroups, Zoo Biol
appetite, weight, hydration, mobility, daily activity, and 20:501–515, 2001.
9. Noon C: Resocialization of a group of ex-laboratory chimpan-
other objective criteria for assessing the quality of life are
zees, Pan troglodytes, J Med Primatol 20:375–381, 1991.
important but not definitive. For example, if an animal 10. Fritz J, Howell S: Captive chimpanzee social group formation. In
in renal failure responds to 20 mg megestrol acetate, an Brent L, editor: The care and management of captive chimpanzees,
appetite stimulant, how should the end-of-life benchmarks San Antonio, TX, 2001, American Society of Primatologists, pp
be adjusted? Do chimpanzees that develop bilateral leg 173–203.
paresis from degenerative disc disease but are mobile with 11. Bloomsmith MA, Baker KC: Social management of captive
ramps and straps and interactive with family members be chimpanzees. In Brent L, editor: The care and management of
considered too compromised? How extensive and intensive captive chimpanzees, San Antonio, TX, 2001, American Society
should the veterinary care of geriatric animals be in zoos, of Primatologists, pp 205–242.
laboratories, or sanctuaries? Electric cardioversion using an 12. Goodall J: The chimpanzees of Gombe: patterns of behavior,
automated external defibrillator successfully revived three Belknap Press of Harvard University, Cambridge, MA, 1986.
13. Botero M, MacDonald SE, Miller RS: Anxiety-related behavior
chimpanzees. One of the chimps, a geriatric male, died
of orphan chimpanzees (Pan Troglodytes schwein furthii) at Gombe
of a fatal arrhythmia 2 years later. The other two female National Park, Tanzania, Primates 54:21–26, 2013.
chimpanzees are still alive and healthy. 14. Brune M, Brune-Cohrs U, McGrew WC, et al: Psychopathol-
Veterinarians should act as the patient’s advocate and ogy in great apes: concepts, treatment options and possible
even more so as they decline. Some veterinarians suggest homologies to human psychiatric disorders, Neurosci Biobehav
geriatric animals may need more frequent physical examina- Rev 30:1246–1259, 2006.
tions, but there is also a valid argument against sedation 15. Bystritsky A, Khalsa SS, Cameron ME, et al: Current diagnosis
due to a higher risk-to-benefit ratio.43 The development of and treatment of anxiety disorders, P T 38(1):30–57, 2013.
noninvasive diagnostic techniques might be more advanta- 16. Pappadopulos E, Woolston S, Chait A, et al: Pharmacotherapy
geous for managing the aging population.44,45 of aggression in children and adolescents: efficacy and effect size,
Veterinarians should remain respectful of the strong per- J Can Acad Child Adolesc Psychiatry 15(1):27–39, 2006.
17. Sorkin EM, Heel RC: Guanfacine: a review of its pharmacody-
sonal bonds and the considerable knowledge of dedicated
namic and pharmacokinetic properties, and therapeutic efficacy
caregivers and include them in the process when making in the treatment of hypertension, Drugs 31:301–336, 1986.
end-of-life and euthanasia decisions. Finally, sanctuaries 18. Findling RL, McNamara NK, Branicky LA, et al: A double-blind
that care for chimpanzees should never underestimate pilot study of risperidone in the treatment of conduct disorder, J
the compassion, protection, and comfort the chimpanzee Am Acad Child Adolesc Psychiatry 39(4):509–516, 2000.
family provides for their failing companions (see also 19. LeBlanc JC, Binder CE, Armenteros JL, et al: Risperidone reduces
Chapter 15). aggression in boys with a disruptive behaviour disorder and below
average intelligence quotient: analysis of two placebo-controlled
randomized trials, Int Clin Psychopharmacol 2(5):275–283, 2005.
20. Turgay A, Binder C, Snyder R, et al: Long-term safety and
References efficacy of risperidone for the treatment of disruptive behavior
disorders in children with subaverage IQ, Pediatrics 110(3):e34,
1. Association of Zoos & Aquariums: Species survival plan® (SSP) 2002.
program handbook, Silver Spring, MD, 2014, Association of Zoos 21. Sleeman J: Great apes. In West G, Heard D, Caulkett N, editors:
& Aquariums. Zoo animal and wildlife immobilization and anesthesia, Ames, IA,
2. Institute of Medicine, Committee on the Use of Chimpanzees in 2007, Blackwell Publishing Professional, pp 387–394.
Biomedical and Behavioral Research: Chimpanzees in biomedical 22. Cerveny S, Sleeman J: Great apes. In West G, Heard D, Caulkett
and behavioral research: assessing the necessity, Washington, DC, N, editors: Zoo animal and wildlife immobilization and anesthesia,
2011, National Academies Press. ed 2, Ames, IA, 2014, Wiley-Blackwell, pp 573–584.
3. ChimpCare.org. Available at: https://readtiger.com/www 23. Horne WA: Primate anesthesia, Veterinary Clinics of North
.chimpcare.org/. America, Exotic animal practice 4(1):239–266, 2001.
4. Freeman HD, Ross SR: The impact of atypical early histories on 24. Murphy HW: Great apes. In Miller RE, Fowler ME, editors:
pet or performer chimpanzees, PeerJ 2:e579, 2014. Fowler’s zoo and wild animal medicine (vol 8). 2015, Saunders,
5. Porton I, Niebruegge K: The changing role of hand rearing pp 336–354.
in zoo-based primate breeding programs. In Sackett G, Rup- 25. Naples LM, Langan JN, Kearns KS: Comparison of the anes-
penthal G, editors: Nursery rearing of nonhuman primates in the thetic effects of oral transmucosal versus injectable medetomidine
21st century, New York, 2005, Kluwer Academic Publishers, pp in combination with tiletamine-zolazepam for immobilization
21–32. of chimpanzees (Pan troglodytes), J Zoo Wildl Med 41(1):50–62,
6. Kalcher E, Franz C, Crailsheim K, et al: Differential onset of 2010.
infantile deprivation produces distinctive long-term effects in 26. Gamble KC, Backues KA: Common Chimpanzee Preventative
adult ex-laboratory chimpanzees (Pan troglodytes), Dev Psychobiol Health Program: Common chimpanzee (Pan troglodytes) preventa-
50(8):777–788, 2008. tive veterinary program for zoos. [www.aazv.org], March 2006.
7. Global Federation of Animal Sanctuaries: Standards for Great 27. Parasitic Diseases of Nonhuman Primates - Exotic and Lab
Ape Sanctuaries, 2013. oratory. http://www.merckvetmanual.com/exotic-and-laboratory
580 SE C T I O N 16 Great Apes
581
582 SE C T I O N 16 Great Apes
• Figure 82.1 By focusing on collaboration and information sharing, zoos and ape caregivers do not
need to “reinvent the wheel” of establishing a network of subject matter experts, diagnostic approaches,
and treatment methods in the care of their great apes.
areas of fibrosis extending through the cardiac chamber treatment of both systolic and diastolic hypertension has
walls. Left ventricular hypertrophy (LVH) is a common been shown to reduce the risk of heart failure.16 Echo-
finding and some apes will progress to have dilated and cardiographic evidence of concentric LVH and systemic
enlarged hearts in end-stage heart failure.3 In humans, the changes in affected apes, similar to those seen in humans
association between sudden cardiac death due to myocardial with hypertension, have strongly implicated hypertension
fibrosis and fatal arrhythmias may be similar to what is seen as a factor in great ape CVD.10,17
in chimpanzees with interstitial myocardial fibrosis, cardiac Defining BP reference ranges for adult, healthy great
arrhythmias, and sudden cardiac death.13,15 apes has been logistically challenging. Historically, BP
The GAHP has developed clinical assessment categories, measurements were attained only from anesthetized great
using ejection fraction (EF) and assessment of functional apes and the effects of various anesthetic agents on BP have
capacity of the heart to determine disease severity. The not been systematically analyzed. Attaining a BP reading
majority of affected apes have been broadly categorized as soon as possible after anesthetic induction and before
as follows: apes with LVH and intact systolic function, commencing inhalant anesthetics, if not using alpha-2
apes with LVH and systolic dysfunction, and apes with a agonists, may give the most accurate BP in an anesthetized
dilated cardiomyopathy phenotype. Aortic dissections are ape. By human standards, consistent systolic readings of
the second leading cause of cardiovascular-related deaths greater than 140 mm Hg or diastolic readings ≥90 mm Hg
in gorillas and bonobos and typically result in acute col- fit the definition of hypertension.18 In chimpanzees, obesity
lapse and death.10,14 Other significant findings (but not as has been shown to be a risk factor for the development
frequently seen) have been aortic root dilation, left atrial of systolic hypertension in females, and increasing age is
enlargement, thromboembolism, right-sided enlargement, a risk factor for development of diastolic hypertension in
arrhythmogenic right ventricular dysplasia/cardiomyopathy, both sexes.19
pulmonary hypertension, inflammatory heart disease, and Attempts to define BP ranges in nonanesthetized great
pericardial effusions. Valvular regurgitation is not typically apes are underway. Generating individual “baselines” for BP
clinically significant. Etiologic factors contributing to ape by consistently attaining BP readings over time may help to
CVD are yet unknown. provide an ancillary tool to monitor great apes for hyperten-
sion. This information can be used to detect changes in
Blood Pressure and Hypertension BP over time and during CVD treatment regimes. Until
BP reference ranges can be developed, it would appear
In humans, elevated blood pressure (BP) is a major risk reasonable that animals with systolic BP consistently above
factor for the development of heart failure, and long-term 160 mm Hg be treated for hypertension.18,19
CHAPTER 82 Update on the Great Ape Heart Project 583
potentially too high and, until more research can be done, Echocardiograms
it is prudent to monitor great ape dietary sodium intake
closely.27 Standardization and accuracy of echocardiographic exam-
inations done in great apes have been essential in detect-
Metabolic Syndrome ing and monitoring great ape CVD, and the GAHP has
developed standardized guidelines for great ape echocar-
In humans, MS is a risk factor associated with cardiomy- diography. A great ape’s size, positioning, and conforma-
opathy and is defined as the presence of at least three of the tion may all affect imaging. It is generally recommended
following: obesity, increased serum triglycerides, reduced to perform echocardiography on anesthetized apes placed
HDL, hypertension, increased fasting glucose, and increased in left lateral recumbency, with the left arm extended cra-
serum insulin levels.21 More research into the role of MS nially (Fig. 82.3).
in ape CVD is needed. In one study done on 16 geriatric Accurate and complete examinations require a skilled
female chimpanzees, 43.8% of the animals met the criteria examiner and consist of a comprehensive, two-dimensional
for MS. Of these animals, 81.2% had some type of CVD.21 transthoracic echocardiogram with Doppler color flow study
capabilities, and all GAHP recommended measurements
Cardiac Health Monitoring stored as DICOM (Digital Imaging and Communications
in Medicine).29,30 While it is possible to review measure-
Echocardiography provides the most practical, clinically ments saved as movie and jpg files, DICOM standard
relevant, and accurate assessment of cardiac functionality, capabilities are the gold standard, especially if participation
valve anatomy, chamber sizes, and ventricular mass and is a in the GAHP is to be maximized. This capability allows
critical tool used for diagnosing CVD in great apes.28 Con- for postprocessing measurements and assessment of images,
siderable expertise is needed in order to obtain a diagnostic ensuring data validity and remote storage capability.
echocardiogram. For this reason, the GAHP strongly rec-
ommends that institutions housing great apes establish rela- Echocardiograms on Nonanesthetized Great Apes
tionships with local cardiologists and echocardiographers. The GAHP has been able to utilize advances in animal
training to encourage cardiovascular monitoring in great
Performing Cardiac Examinations apes without the aid of anesthesia. The advantages of
performing echocardiography and BP monitoring on non-
Echocardiographic assessments should be done on great anesthetized apes include less frequent anesthetic episodes,
apes during every anesthetized examination once the ape more frequent monitoring, and lack of anesthetic effects on
reaches adulthood. At a minimum, adult echocardiograms the cardiovascular system. The disadvantages include train-
should be done every 2–3 years on animals with normal ing time and logistics, risk to the trainer and equipment,
cardiac health. Once an animal is determined to be affected less thorough echocardiograms, and a missed opportunity
by CVD, a risk analysis should be used to aid in determin- to do a complete physical examination. Unfortunately,
ing anesthesia and examination frequency (Table 82.1). echocardiograms done on awake animals may take several
sessions to obtain all the necessary measurements. Therefore,
while not ideal, the GAHP recommends that measurements
from training sessions obtained within a 30-day period be
submitted as one examination.
TABLE Great Ape Heart Project Recommendations
82.1 for Frequency of Blood Pressure and
Echocardiography Exam Based on Age
and Health Status
Age/Health Status Frequency of Exams
Neonate Opportunistically if a neonate has
to be removed from the dam
for any reason, a neonatal
exam should be done.
9 years Baseline exam
10–20 years Every 3–5 years
>20 years Every 2–3 years
Animals with cardiac Examination frequency should be
disease determined on a case-by-case
basis in order to monitor and
manage treatment. • Figure 82.3 An anesthetized gorilla (Gorilla gorilla) shown in left
lateral recumbency.
CHAPTER 82 Update on the Great Ape Heart Project 585
28 chimpanzees, cTnI levels were found to have a predictive clinically relevant diagnostic and treatment criteria for
value for CVD disease in cases of advanced/severe cardiac future evaluations.
disease, but the levels were not predictive in cases of mild
to moderate severity.43 The authors of this study concluded Acknowledgments
that any observed value of cTnI above the detection thresh-
old of the assay used (0.20 ng/mL) should be treated as an The GAHP is supported by the Institute of Museum
indicator of potential CVD in chimpanzees, although a and Library Services (Grants LG-26-12-0526-12 and
limitation of this study was that the cTnI assay used had MG-30-15-0035-15) and Zoo Atlanta. We are grateful for
an analytic sensitivity above the threshold for humans and the support and involvement of the Ape Taxon Advisory
veterinary species; therefore some chimpanzees with heart Group, the individual ape SSPs (veterinary, pathology, and
disease may not have been detected.43 nutrition advisors), the veterinary and medical cardiologists
Biomarkers to detect extracellular matrix remodeling and and sonographers who generously donate their time, and
fibrogenesis resulting in myocardial fibrosis formation and various staff at all participating institutions.
turnover in great apes have shown promise and are currently
under investigation.45
References
Treating Cardiovascular Disease in 1. Seiler BM, Dick EJ, Jr, Guardado-Mendoza R, et al: Spontane-
ous heart disease in the adult chimpanzee (Pan troglodytes), J Med
Great Apes Primatol 38:51–58, 2009.
2. Lammey ML, Lee DR, Ely JJ, et al: Sudden cardiac death in 13
Standard pharmacologic therapies for cardiomyopathies captive chimpanzees (Pan troglodytes), J Med Primatol 37(Suppl
including LV dysfunction in humans include beta- 1):39–43, 2008.
adrenergic blockade, renin-angiotensin-aldosterone system 3. McManamon R, Lowenstine L: Cardiovascular disease in great
antagonism, and diuretics as necessary for congestive heart apes. In Miller RE, Fowler ME, editors: Fowler’s zoo and wild
failure.16 To date, most of the GAHP advisory experi- animal medicine current therapy, ed 1, 2012, pp 408–415.
ence in the great apes has been with beta-blockade and 4. Meehan T, Lowenstine L: Causes of mortality in captive lowland
angiotensin-converting enzyme (ACE) inhibition, with gorillas: a survey of the SSP population. In Proceedings of the
other pharmacologic agents used on a more sporadic basis. American Association of Zoo Veterinarians Annual Conference,
It is worth emphasizing that no clinical studies have 1994, pp 190–192.
5. Lowenstine L, McManamon R, Bonar CJ, et al: Preliminary
been performed on the efficacy, safety, or pharmacokinetics
results of a survey of United States and Canadian orangutan
of any cardiovascular therapeutics in apes, so recommenda- mortalities in the North American SSP population from 1980
tions are based purely on experience with them in apes over to March 2008. In Proceedings of the American Association of
the course of the GAHP studies, as well as hypothetical Zoo Veterinarians Conference, 2008.
assumptions that apes would react in much the same way 6. Gamble KC: Pathologic review of the chimpanzee (Pan troglo-
as humans do to these drugs. Attending veterinarians must dytes): 1990–2003. In Proceedings of the American Association
use their best judgment when prescribing these drugs as of Zoo Veterinarians/American Association of Wildlife Veterinar-
“off-label” medications and should monitor animals very ians Conference, 2004, pp 561–566.
closely for any adverse side effects. 7. Clyde VL, Roth L, Bell B, et al: Cardiac and gestational ultra-
Thromboembolism and cerebral infarcts have been sound parameters in nonanesthetized bonobos (Pan paniscus). In
documented in the great apes.21,46,47 In these cases, aspirin Proceedings of the American Association of Zoo Veterinarians
Annual Meeting, 2002, pp 365–368.
therapy has been recommended.21
8. Varki N, Anderson D, Herndon JG, et al: Heart disease is
common in humans and chimpanzees, but is caused by different
Postmortem Cardiac Evaluations pathological processes, Evol Appl 2:101–112, 2009.
9. Junge RE, Mezei LE, Muhlbauer MC, et al: Cardiovascular
The pathology group of the GAHP has collected and evaluation of lowland gorillas, J Am Vet Med Assoc 212:413–415,
reviewed available necropsy and histopathology reports 1998.
from captive apes. All information collected is reviewed 10. Lowenstine LJ, McManamon R, Terio KA: Comparative pathol-
and entered into the GAHP database and is analyzed for ogy of aging great apes, Vet Pathol 53:250–276, 2015.
antemortem clinical correlations, inter and intra-specific 11. Kambale ES, Ramer JC, Gilardi K, et al: Cardiovascular and
taxon trends, and disease classifications. Novel postmortem hepatic disease in wild Eastern Lowland Gorillas (Gorilla beringei
cardiac tissue collection and evaluation techniques for great graueri). In Proceedings of the American Association of Zoo
Veterinarians Annual Conference, 2014, p 115.
ape necropsies have been developed, and these protocols
12. Terio KA, Kinsel MJ, Raphael J, et al: Pathologic lesions in chim-
have established a new “best practices” approach to ape panzees (Pan trogylodytes schweinfurthii) from Gombe National
heart evaluation that is more closely aligned with tech- Park, Tanzania, 2004–2010, J Zoo Wildl Med 42:597–607,
niques used in human cardiac autopsies.48 These guidelines 2011.
may be found at www.greatapeheartproject.org. Database 13. Lammey ML, Baskin GB, Gigliotti AP, et al: Interstitial myocar-
inquiry coupled with simultaneous multipathologist dial fibrosis in a captive chimpanzee (Pan troglodytes) population,
review will allow the GAHP to establish more precise and Comp Med 58:389–394, 2008.
CHAPTER 82 Update on the Great Ape Heart Project 587
14. Kenny DE, Cambre RC, Alvarado TP, et al: Aortic dissection: an 32. Magden ER, Sleeper MM, Buchl SJ, et al: Use of an implant-
important cardiovascular disease in captive gorillas (Gorilla gorilla able loop recorder in a chimpanzee (Pan troglodytes) to monitor
gorilla), J Zoo Wildl Med 25:561–568, 1994. cardiac arrhythmias and assess the effects of acupuncture and
15. Doane CJ, Lee DR, Sleeper MM: Electrocardiogram abnor- laser therapy, Comp Med 66:52–58, 2016.
malities in captive chimpanzees (Pan troglodytes), Comp Med 33. Duncan AE, Kutinsky I, Murray S, et al: Use of implantable loop
56:512–518, 2006. recorders to monitor for cardiac disease in captive gorillas (Gorilla
16. Yancy CW, Jessup M, Bozkurt B, et al: 2013 ACCF/AHA guide- gorilla gorilla), Proceedings of the American Association of Zoo
line for the management of heart failure: executive summary: Veterinarians Annual Conference, 2014.
a report of the American College of Cardiology Foundation/ 34. West G, Heard D, Caulkett N: Zoo animal and wildlife immobi-
American Heart Association Task Force on practice guidelines, lization and anesthesia, ed 2, 2014, pp 573–584.
Circulation 128:1810–1852, 2013. 35. Adami C, Wenker C, Hoby S, et al: Evaluation of effectiveness,
17. Grim CE, Highland MA, Beehler L, et al: Familial hypertension safety and reliability of intramuscular medetomidine-ketamine
and stroke in bonobo chimpanzees (Pan paniscus). In Proceedings for captive great apes, Vet Rec 171:196, 2012.
of the American Heart Association, Hypertension E127–E127, 36. Horne WA, Norton TM, Loomis MR: Cardiopulmonary effects
2010. of medetomidine-ketamine-isoflurane anesthesia in the gorilla
18. Ely JJ, Zavaskis T, Lammey ML, et al: Blood pressure reference (Gorilla gorilla) and chimpanzee (Pan troglodytes). In Proceed-
intervals for healthy adult chimpanzees (Pan troglodytes), J Med ings of the American Association of Zoo Veterinarians Annual
Primatol 40:171–180, 2011. Conference, 1997, pp 140–142.
19. Ely JJ, Zavaskis T, Lammey ML: Hypertension increases with 37. Talke P, Lobo E, Brown R: Systemically administered
aging and obesity in chimpanzees (Pan troglodytes), Zoo Biol α2-agonist-induced peripheral vasoconstriction in humans,
32:79–87, 2013. Anesthesiology 99:65–70, 2003.
20. Popovich DG, Jenkins DJ, Kendall CW, et al: The western 38. Naples LM, Langan JN, Kearns KS: Comparison of the anes-
lowland gorilla diet has implications for the health of humans thetic effects of oral transmucosal versus injectable medetomidine
and other hominoids, J Nutr 127:2000–2005, 1997. in combination with tiletamine-zolazepam for immobilization
21. Nunamaker EA, Lee DR, Lammey ML: Chronic diseases in of chimpanzees (Pan troglodytes), J Zoo Wildl Med 41:50–62,
captive geriatric female chimpanzees (Pan troglodytes), Comp Med 2010.
62:131–136, 2012. 39. Pypendop BH, Verstegen JP: Hemodynamic effects of medeto-
22. Seng CF, Meng GY, Abdullah R: A comparative study of blood midine in the dog: a dose titration study, Vet Surg 27:612–622,
lipids and stress levels between captive and semi-captive orang- 1998.
utans (Pongo pygmaeus) under tropical conditions. In Proceedings 40. Napier JE, Kutinsky IB, Armstrong DL, et al: Evaluating echo-
of the American Association of Zoo Veterinarians/American cardiogram and indirect blood pressure results in male western
Association of Wildlife Veterinarians/American Zoo Association/ lowland gorillas (Gorilla gorilla gorilla) during three phases of an
Nutrition Advisory Group Joint Conference, 2007. anesthetic protocol, J Zoo Wildl Med 44:875–881, 2013.
23. Schmidt DA, Ellersieck MR, Cranfield MR, et al: Cholesterol 41. Lopez del Rio PR, Sanchez C, Stembridge M, et al: Comparison
values in free-ranging gorillas (Gorilla gorilla gorilla and Gorilla of indirect blood pressure values in chimpanzees (Pan troglodytes)
beringei) and Bornean orangutans (Pongo pygmaeus), J Zoo Wildl anesthetized with two anesthetic protocols: tiletamine-zolazepam
Med 37:292–300, 2006. and titetamine-zolazepam-medetomidine. In Proceedings of the
24. Baitchman EJ, Calle PP, Clippinger TL, et al: Preliminary evalu- American Association of Zoo Veterinarians Annual Conference,
ation of blood lipid profiles in captive western lowland gorillas 2014.
(Gorilla gorilla gorilla), J Zoo Wildl Med 37:126–129, 2006. 42. Raizada A, Bhandari S, Khan MA, et al: Brain type natriuretic
25. Mente A, O’Donnell M, Rangarajan S, et al: Associations of peptide (BNP) - A marker of new millennium in diagnosis of
urinary sodium excretion with cardiovascular events in individu- congestive heart failure, Indian J Clin Biochem 22:4–9, 2007.
als with and without hypertension: a pooled analysis of data from 43. Ely JJ, Zavaskis T, Lammey ML, et al: Association of brain-
four studies, Lancet 388:465–475, 2016. type natriuretic protein and cardiac troponin I with incipient
26. Denton D, Weisinger R, Mundy NI, et al: The effect of cardiovascular disease in chimpanzees (Pan troglodytes), Comp
increased salt intake on blood pressure of chimpanzees, Nat Med Med 61:163–169, 2011.
1:1009–1016, 1995. 44. Sharma S, Jackson PG, Makan J: Cardiac troponins, J Clin Pathol
27. Council NR, Studies DEL, Resources BAN, et al: Nutrient 57:1025–1026, 2004.
requirements of nonhuman primates, Revised ed 2, 2003, National 45. Ely JJ, Bishop MA, Lammey ML, et al: Use of biomarkers of col-
Academies Press. lagen types I and III fibrosis metabolism to detect cardiovascular
28. Lang RM, Badano LP, Mor-Avi V, et al: Recommendations for and renal disease in chimpanzees (Pan troglodytes), Comp Med
cardiac chamber quantification by echocardiography in adults: 60:154–158, 2010.
an update from the American Society of Echocardiography and 46. Weisenberg E, Snook S, Letcher J: Sudden death in an obese
the European Association of Cardiovascular Imaging, J Am Soc orangutan with hypertensive heart disease and a history of stroke
Echocardiogr 28:1–39, 2015. (abstract). Proceedings of the American Association of Zoo
29. Shave R, Oxborough D, Somauroo J, et al: Echocardiographic Veterinarians Annual Conference, 1991.
assessment of cardiac structure and function in great apes: a 47. Trupkiewicz JG, McNamara TS, Weidenham KM, et al: Cerebral
practical guide, Int Zoo Yearb 48:218–233, 2014. infarction associated with coarctation of the aorta in a lowland
30. Sleeper MM, Drobatz K, Lee DR, et al: Echocardiographic gorilla (Gorilla gorilla gorilla), J Zoo Wildl Med 26:123–131,
parameters of clinically normal adult captive chimpanzees (Pan 1995.
troglodytes), J Am Vet Med Assoc 244:956–960, 2014. 48. Terio KA, McManamon R, Ellis AE, et al: Best practices for the
31. Lammey ML, Jackson R, Ely JJ, et al: Use of an implantable cardiac necropsy: tips for clinicians (and pathologists) perform-
loop recorder in the investigation of arrhythmias in adult captive ing ape necropsies. In Proceedings of the American Association
chimpanzees (Pan troglodytes), Comp Med 61:71–75, 2011. of Zoo Veterinarians Annual Conference, 2014, pp 88–90.
83
Great Ape Nutrition
DEBRA A. SCHMIDT AND MICHELLE E. SHAW
Wild Diets and Digestive Physiology (Dorylus rubellus, Oecophylla longinoda, Apis spp.) are
commonly found in fecal samples.12,16–19 Bonobos have
Diets of free-ranging great apes are largely plant-based. Rela- also been reported to consume animal matter includ-
tive inclusions of types of plants and animal material have ing rodents (order Rodentia), duikers (Cephalophus sp.),
resulted in their classification by feeding strategy. Captive monkeys (Cercopithecus wolfi, Cercopithecus ascanius
diets have a tendency to reflect human-assigned feeding schmidti), and invertebrates (Macrotermes, Dorylus, and
strategies rather than the capability each species has to digest Apis).20–24 However, the protein contribution is insignif-
the nutrient levels in their natural diet. Adaptations in the icant so the behavior may be primarily for social bene-
gastrointestinal (GI) tract of each species ascertain their fits.25 There are also a few reports of carnivorous activity
relative ability to digest fiber and should be taken into by orangutans, including the consumption of slow lorises
account. (Nycticebus spp.), but animal matter is considered to com-
Gorillas (Gorilla spp.) are considered the most herbivo- prise an even smaller portion of the diet than it does in
rous of all great apes with a natural diet of plant leaves, chimpanzees.26–28
stems, pith, bark, roots, flowers, and fruit.1–3 Orangutans
(Pongo spp.) have a very similar GI tract and therefore Body Weights
comparable digestive capabilities. Gorillas and orangutans
have a lengthier small intestine and a more voluminous large In captivity, all individuals should have a goal of weight
intestine compared to other great apes indicative of their loss, gain, or maintenance. Ideally, body condition scoring
enhanced fermentative capabilities, which are more similar in conjunction with body weight assessments should be
to those of a horse than a human (Fig. 83.1). Orangutans performed regularly to determine the optimal weight for
are often classified as frugivores, due to their preference for each individual. Body weights for chimpanzees in the wild
fruit when it is available, but they have been reported to eat a range from 49–80 kg for males and 40–68 kg for females.29
variety of foods including fruit, leaves, nonleafy vegetation, Free-ranging bonobo weights are less available but are listed
inner bark, flowers, and insects.4–8 Gorillas and orangutans as 37–61 kg for males and 27–38 kg for females.30 In the
have been observed eating decaying wood and clay-rich wild, male orangutans weigh 80–91 kg, while females weigh
soil respectively.9,10 These may be maintaining a healthy 33–45 kg; therefore it is not unusual for full-flanged males
gut environment by contributing to the gut microbiome to be more than twice the size of females.31 Free-ranging
or adsorbing toxins. male and female gorillas have weight ranges of 130–218
The GI tracts of chimpanzees (Pan troglodytes) (see Fig. and 68–74 kg respectively, although wild silverbacks were
83.1) and bonobos (Pan paniscus) are very similar to each found to eat only 1.3 times the amount consumed by adult
other with a relatively short small intestine and a longer females.32,33
large intestine and colon with comparatively lower fiber fer-
mentative capacity than orangutans and gorillas, although
they do have the ability to ferment fiber for energy produc- Recommended Diet Plan
tion.11 Chimpanzees consume a diet consisting primarily of Water
fruit, but they also eat leaves, pith, seeds, flowers, insects,
and meat leading to the classification of an omnivorous Water is the most important nutrient for any animal; an
frugivore.12 Bonobos consume a diet consisting mostly of animal will die from dehydration before it will die from any
plant material with the majority of it being fruit, but they other nutrient deficiency. Apes should have unlimited access
also consume leaves, shoots, stems, flowers, and pith.13–15 to fresh, clean drinking water at all times. It is important
Chimpanzees have been noted to eat bushbuck to clean water containers or devices daily to prevent illness.
(Tragelaphus scriptus), bushpig (Potamochoerus larvatus), Dehydrated animals are more likely to become constipated
red colobus (Procolobus sp.), and animal parts and insects and have dry skin; in humans it can also lead to kidney
588
CHAPTER 83 Great Ape Nutrition 589
0 cm 50
0 cm 25
0 cm 30
• Figure 83.1 Comparative digestive anatomy of the zebra (top left), orangutan (top right), chimpanzee
(bottom left) and human (bottom right). Stevens CE, Hume ID, editors: Comparative physiology of the
vertebrate digestive system, ed 2, Cambridge, UK, 1995, Cambridge University Press.)
stones and liver, muscle, and joint damage, problems that contains appropriate concentrations of vitamins, minerals,
may also affect great apes. fats, and protein for primates. Using a product specifically
formulated for use with primates as a basis for the diet
Kibble/Pellets is highly encouraged; however, a pellet formulated for
horses may also be used and may be more economical and
Some feed manufacturers (e.g., Mazuri or NutraZu, PMI available (Shaw, personal communication). These products
Nutritional International, Brentwood, Missouri; Marion contain higher concentrations of fiber than produce items
Zoological, Plymouth, Minnesota) produce a dry kibble may provide and supplemental micronutrients that would
(biscuit) specifically formulated for primates. The feed otherwise be deficient. Although they may not be as
590 SE C T I O N 16 Great Apes
palatable to a primate as produce, they may be blended than twice as many calories as protein or carbohydrates.
with more palatable items to provide the basis for a healthy, Lower-fat seeds, such as pumpkin, hemp, and sesame seeds,
balanced great ape diet. are healthier and provide a good source of fiber, healthy fat,
vitamins, and minerals.
Browse
Vitamin and Mineral Supplementation
It is impossible to match the fiber concentrations consumed
by wild apes using only produce commercially grown for Providing a variety of nutritionally appropriate foods will
human consumption. Leafy tree branches (nontoxic browse) help reduce the risk of deficiencies. When apes receive
and dried alfalfa (leafy material and stems) are excellent and consume a well-balanced diet based on a formulated
feed items to increase fiber in the diet. If these items are pellet, supplementation with vitamins and minerals is not
readily available and reliably consumed, then they should necessary. For those animals that do not have access to
be offered as much as possible throughout the year. When primate kibble or a commercial horse pellet, a complete
browse is available, feeding time is increased and regurgita- multivitamin formulated for adult humans is suggested
tion and reingestion (R&R) are reduced.34 Some animal to avoid deficiencies in micronutrients. It is possible to
care facilities have partnered with utility companies to use formulate a balanced diet without additional supplementa-
the tree branches that are trimmed away from utility lines tion, but the complete nutrient profile of all available food
as it is unlikely that trees along utility lines are treated with items must be known to confirm that the overall diet is
pesticides or herbicides. This partnership provides browse adequate. When an ape has been confirmed to be pregnant
for the animals and offers a favorable image for the utility or is lactating, supplementing them with a prenatal vitamin
company as a result of helping feed endangered animals. If may be considered.
browse or alfalfa is not available all year, the amount of leafy Access to natural, unfiltered sunlight for at least 30
green vegetables may be increased. minutes daily is beneficial for all life stages of apes, but is
most critical for young animals.36,37 Because vitamin D3 is
Vegetables not transported well through dam’s milk, mother-reared
infants who do not have routine access to natural (outdoor)
In a great ape diet, 85%–90% will be made up of browse unfiltered sunlight (not through glass, skylights or mesh)
and leafy green and low-starch vegetables. Large amounts of will benefit from an oral, supplemental dose of vitamin D3
these vegetables will create a feeling of fullness and increase to support appropriate bone growth and development.38 A
the amount of foraging time without adding a lot of calories vitamin D3 supplement for human, breastfed infants works
to the diet. Offer vegetables with the peels and cores intact well with great apes.
when possible as they provide potential sources of fiber
and may provide a source of enrichment by requiring the Animal Products
ape to process food before consuming it. However, items
that are potentially toxic, such as avocado pits and skins, Feeding apes animal products is not recommended as they
or fibrous skins or seeds that may be a choking hazard in are difficult to digest, promote obesity, and may increase the
younger animals, should not be offered to apes; always err incidence of R&R, particularly in gorillas, orangutans, and
on the side of caution. chimps.39 There are a few exceptions to this rule for hand-
Cooked produce items, especially root vegetables, raised or compromised individuals. Dairy-based human
contain more easily digested sugars than uncooked items.35 infant formulas supplemented with omega fatty acids are
All produce should be offered raw to minimize the amount recommended when hand-raising great apes as human milk
of easily digested sugars and only cooked when necessary. is similar in composition to great ape milk.40 Older animals
For example, animals with dental problems or older animals or those with health concerns that are having difficulty
in poor body condition may benefit from having some maintaining weight may be supplemented with hardboiled
items cooked to improve consumption. eggs a few times per week to provide a high-quality protein
source.
Seeds, Nuts, Whole Grains, Pulses/Legumes
Fruit
Locally available seeds and nuts can be used to balance
and/or provide variety to a great ape diet. Whole grains High levels of fruit are reported in free-ranging ape diets;
(e.g., millet, barley, wheat, brown rice, and oats), legumes however, the fruits in the wild are significantly higher in fiber
(e.g., alfalfa, peas, soybeans, and beans such as kidney, and lower in readily absorbable sugars than fruits and other
pinto, adzuki, mung, and broad beans), and lentils are produce cultivated for human consumption and should not
a good source of protein, fiber, and minerals. High-fat be considered nutritionally equivalent.41 The wild varieties
seeds (e.g., sunflower), nuts, and high-fat legumes (e.g., are more nutritionally similar to the seed-bearing parts of
peanuts), should be used in moderation, especially with plants, such as eggplant, zucchini, and capsicum, which are
overweight animals. On a gram-per-gram basis, fat has more botanically defined as fruit and are more often considered
CHAPTER 83 Great Ape Nutrition 591
The proportions listed in Table 83.1 are recommended when Phosphorus, % 0.6
feeding apes and meet the recommendations established for Magnesium, % 0.08
the Nutrient Requirements of Nonhuman Primates (Table Potassium, % 0.4
83.2).43 Though apes in the wild consume higher concentra-
tions of fiber than those recommended, it is very difficult to Sodium, % 0.2
mimic those high fiber concentrations in a zoo setting, and Chloride, % 0.2
for this reason, all four great ape species are fed similar diets. Iron, mg/kg 100
Examples of amounts to offer when using biscuit/pellet/
kibble, relative to animal body weight, are listed in Table Copper, mg/kg 20
83.3. Examples of amounts to offer when the diet is based Manganese, mg/kg 20
on tofu and seeds/nuts/legumes are listed in Table 83.4. Zinc, mg/kg 100
Tofu may increase R&R in some animals. If this behavior
occurs, replace tofu with a variety of seeds, nuts, and pulses. Iodine, mg/kg 0.35
When feeding a diet based on tofu, seeds, nuts, whole Selenium, mg/kg 0.3
grains, and pulses, a vitamin and mineral supplement tablet Vitamin A, IU/kg 8000
formulated for adult or juvenile humans is recommended,
based on the age of the animal. Vitamin D3, IU/kg 2500
Vitamin E, mg/kg 100
Vitamin K, mg/kg 0.5
Thiamin (B1), mg/kg 3.0
Riboflavin (B2), mg/kg 4.0
Pantothenic acid, mg/kg 12.0
TABLE Recommended Ape Diet Proportions by
83.1 Weight (As-Fed Basis) Niacin, mg/kg 25.0
Vitamin B6, mg/kg 4.0
Tofu and
Seeds/Nuts/ Biotin, mg/kg 0.2
Diet Item Kibble/Pellets (%) Legumes (%) Folacin, mg/kg 4.0
TABLE
83.3 Kibble/Pellet and Vegetable Diet Amounts Based on Animal Weight (As-Fed Basis)
Animal Weight
TABLE
83.4 Tofu, Seeds/Nuts/Legume and Vegetable Diet Amounts Based on Animal Weight
Animal Weight
the following equation for each food category and uses the items, which can be purchased at a lower cost, to be used
current weight of the animal: and provide enrichment. Creating a weekly, documented
schedule of the rotation may help caretakers maintain the
85 kg ÷ 70 kg = 1.21 × 468 g ( kibble 70 kg animal) variety and rotation.
= 568 g kibble pellets 85 kg animal
122 kg ÷ 110 kg Enrichment
= 1.11 × 8710 g ( leafy greens 110 kg animal) Just providing the animal with additional food does not
= 9660 g leafy greens 122 kg animal meet the definition of enrichment. All food may be classified
as “enrichment” if it is provided in a stimulating manner
that encourages the great ape to exhibit natural behaviors.
Feed Presentation It is also important to quantify and calculate the calories
Group Feeding being offered in enrichment items daily. It is tempting to
use higher-fat nuts and seeds as enrichment feeds due to
When feeding animals in a group, the fastest or most domi- their small size and potential processing requirements, but
nant animal must be prevented from gathering and eating that could cause the apes to consume too many calories.
the most desired items. This can be overcome by target Enrichment and training may be done using the foods
feeding the highly desired, high-calorie foods to animals that are typically part of their diet. In the wild they spend
individually; they may even be used as training rewards. The a majority of their day searching for food; to mimic this,
less calorically dense vegetables and leafy green vegetables their produce can be cut into small pieces and scattered
may be group fed, ensuring a more balanced diet for all throughout the enclosure. It may also be mixed in hay or
apes in the group. hidden in different areas of their habitat. If infant apes
If a variety of produce items are available, every oppor- are present, consider the size of the foods so they do not
tunity should be taken to offer the apes an assortment of create a choking hazard. Foods may also be kept whole and
items throughout the week. More variety is attained when spiked around the exhibit or hung from ropes making them
offering larger amounts of only four or five different items difficult to retrieve. Food items may be diced or pureed
daily and rotating the selection throughout the week than and placed in a tube or crafted concrete termite mound
to offer smaller amounts of the same 20 items daily. This where the apes have to use long, thin sticks to “fish” for
will help to ensure that a few individuals do not always eat the items (like they would for termites). Puzzle feeders are
the more favored items every day. It will also allow seasonal another good way to make them think, expend energy,
CHAPTER 83 Great Ape Nutrition 593
and work for their food. It is always best to follow the diet the animal accustomed to being weighed weekly or every
plan and use only those food items and predetermined other week is ideal. The next step would be to transition the
amounts as enrichment by providing them in novel and animal to one of the above-mentioned diets (pellets/kibble
interesting ways. or tofu/seeds/nuts/ grains if pellets/kibble are not available)
in the appropriate proportions (see Tables 83.3 and 83.4).
Health It is not as important for the animal to consume the listed
amount of food that corresponds to their exact weight.
A recent review of reports published between 1990 and Instead, it is important that the animal consume the major-
2014 identified idiopathic and infectious diseases as well as ity of the diet offered and is not offered so much food that
cardiovascular, respiratory, and GI disorders as the leading the animal may be selective and satiated on only the foods
causes of captive great ape morbidity and mortality.44 Great he or she chooses to eat. Finding the diet that most closely
apes are at serious risk for anthropozoonotic diseases. resembles the ape’s current diet should be considered the
GI and upper respiratory tract disorders are most often starting point. Once the diet quantity has been determined
reported and easily transmitted from caregivers through the and the animal has settled into this new diet, begin reducing
preparation of ape diets. Washing hands frequently during all diet categories by 10%; do not be selective in which
diet preparation should be a top priority regardless of health categories are reduced (i.e., only reducing the categories that
status, and the use of facemasks and gloves is recommended the animal does not like). By reducing all diet categories, the
in higher-risk situations. items will remain in proportion with one another and the
diet offered will remain balanced. Changing an individual’s
Orangutan Air Sacculitis diet in group-housed apes can be particularly challenging.
It is also important not to move quickly with this
Air sacculitis is the most frequently reported respiratory process in an effort to encourage rapid weight loss. This
disorder in captive orangutans.44 It is generally considered is not a healthy way to lose weight; the animal’s daily
to be caused by environmental factors such as dust; however, nutrient needs may not be met, and the metabolism could
there are several nutrition-related factors that may contrib- be altered, making it more difficult to achieve weight loss
ute to this condition. Anything that promotes aspiration goals. Whenever diet changes are made, even to a healthier
of food particles into the air sacs will increase the risk of diet, digestive upset may occur. If an animal has gas or
developing air sacculitis. Obese individuals and those that diarrhea for one day, this is perfectly normal, but if digestive
spend all their time on the ground without climbing or upset is prolonged, no further diet changes should be made
brachiating (which physically stretches out their bodies) until fecal consistency improves. Never change an animal’s
may experience a physical constriction of the air sacs. diet by more than 5%–10% at a time to minimize risk of
Animals that habitually practice R&R are also at increased digestive upset.
risk. This is one reason that any food that stimulates R&R Removing high-sugar items is particularly difficult.
behavior, such as sugary foods, dairy, and tofu, should be When removing sugary foods, the animal may exhibit
restricted and eliminated completely from the diet if R&R signs of withdrawal (e.g., headaches, irritability, frustration,
is observed. nausea). These symptoms are temporary and will usually
Treatment of air sacculitis often involves administering disappear in a few weeks after sugar is removed. The removal
antibiotics, which may disrupt the normal microbial popu- of high-sugar items in primate diets has been found to
lation of the gut.45 To improve gut function after antibiotics reduce negative stereotypic behaviors such as begging and
are administered, animals may be fed feces from a healthy self-mutilation.48 The overall improvements in psychologi-
individual to repopulate the microbiome. This can be cal and physical health are worth a few weeks of symptoms
achieved by adding the feces to oatmeal or vegetable purée. associated with sugar withdrawal.
Additionally, group housed apes may naturally recolonize Weight changes should be achieved in a slow and con-
their GI tract through fecal exposure. trolled manner. Diet and activity levels are the two most
critical components in maintaining animals at appropriate
Obesity weights. Weighing animals frequently allows early detection
of rapid weight changes. Enclosures should also provide
Obesity in humans can lead to a multitude of health-related opportunities for activities such as climbing, swinging,
conditions, including death, high blood pressure, heart and hanging to increase energy expenditure and maintain
disease, cancer, degenerative arthritis, respiratory problems, muscle mass. Weight changes should not exceed 2% of
diabetes, fatty liver disease, and lower fertility in women.46,47 body weight weekly. For example, a 100-kg orangutan,
Obese apes may also be at risk for these problems. determined to be overweight, should have diet changes that
result in a body weight loss of 1–2 kg weekly.
Obesity—Weight Loss Diet
To begin a weight loss diet, it is important to first deter- Obesity—Expending Energy
mine the animal’s current weight and ideally take several Consider using their typical dietary foods to encourage
photographs of the animal from various angles. Getting movement around the enclosure. Think of ways to offer
594 SE C T I O N 16 Great Apes
food that will require apes to climb ropes, move from 8. Vogel E, Alavi S, Utami-Atmoko S, et al: Nutritional ecology
platform to platform, hang upside down, move objects, or of wild Bornean orangutans (Pongo pygmaeus wurmbii) in a peat
stretch high above their heads. Hanging food from ropes swamp habitat: Effects of age, sex and season, Am J Primatol
79:e22618, 2017.
that keeps food just barely in reach will encourage these
9. Rothman JM, Chapman CA, Pell AN: Fiber-bound nitrogen in
behaviors. Cutting vegetables in small pieces and scattering gorilla diets: Implications for estimating dietary protein intake
them around the enclosure requires the animal to move of primates, Am J Primatol 70:690–694, 2008.
around the habitat to gather the food. Hiding food items 10. Mahaney WC, Hancock RGV, Aufreiter S, et al: Bornean orang-
so animals need to search for it will also stimulate more utan geophagy: analysis of ingested and control soils, Environ
activity, which is helpful for weight loss. Creating challenges Geochem Health 38:51–61, 2016.
that require a greater effort to access food may have the 11. Stevens CE, Hume ID: The mammalian gastrointestinal tract.
added benefit of making less-favored items more desirable, In Comparative physiology of the vertebrate digestive system, ed 2,
improving the acceptance of a healthier diet.49 Cambridge, UK, 1995, Cambridge University Press.
To reduce aggression with multiple apes sharing an enclo- 12. Phillips C, McGrew W: Identifying species in chimpanzee (Pan
sure, use the higher-value portion of the diet as a reward troglodytes) feces: a methodological lost cause?, Int J Primatol
34:792–807, 2013.
for training instead of scattering it within the enclosure.
13. Malenky RK, Stiles EW: Distribution of terrestrial herbaceous
Providing food in several smaller meals throughout the day vegetation and its consumption by Pan paniscus in the Lomako
at various times will also encourage more foraging behavior. Forest, Zaire, Am J Primatol 23:153–169, 1991.
14. Malenky RK, Wrangham RD: A quantitative comparison of
Cardiovascular Disease terrestrial herbaceous food consumption by Pan paniscus in the
Lomako Forest, Zaire, and Pan troglodytes in the Kibale Forest,
Although great apes are genetically similar to humans, they Uganda, Am J Primatol 32:1–12, 1994.
develop a very different type of heart disease. Apes do not 15. Hohmann G, Potts K, N’Guessan AK, et al: Plant foods con-
get atherosclerosis with plaque buildup as seen in humans; sumed by Pan: exploring the variation of nutritional ecology
instead, they are frequently diagnosed with fibrosing across Africa, Am J Phys Anthropol 141(3):476–485, 2010.
cardiomyopathy (i.e., scar tissue buildup in the heart) at 16. Phillips C, O’Connell T: Fecal carbon and nitrogen isotopic
analysis as an indicator of diet in Kanyawara chimpanzees, Kibale
necropsy.50–52 Mortality reports cite cardiovascular disease
National Park, Uganda, Am J Phys Anthropol 161:685–697,
as the primary cause of death in captive great apes and 2016.
as a premature cause of death in young, captive apes (see 17. Sommer V, Buba U, Jesus G, et al: Sustained myrmecophagy
Chapter 82).53 in Nigerian chimpanzees: Preferred or fallback food?, Am J Phys
Sodium levels in the diet need to be considered in relation Anthropol 162:328–336, 2017.
to heart health. A current theory exists that hypertension 18. Tappen M, Wrangham R: Recognizing hominoid-modified
plays a role in great ape heart disease. High sodium intake bones; The taphonomy of colobus bones partially digested by
is linked to high blood pressure in humans, and therefore free-ranging chimpanzees in the Kibale Forest, Uganda, Am J
monitoring and minimizing the amount of salt an animal Phys Anthropol 113:217–234, 2000.
consumes is important. 19. Tutin CEG, Fernandez M: Insect-eating by sympatric Lowland
gorillas (Gorilla g. gorilla) and chimpanzees (Pan t. troglodytes) in
the Lopé Reserve, Gabon, Am J Primatol 28:29–40, 1992.
References 20. Kano T: The last ape: pygmy chimpanzee behavior and ecology,
Stanford, CA, 1992, Stanford University Press.
1. Remis MJ: Western lowland gorillas (Gorilla gorilla gorilla) as 21. Malenky RK: Ecological factors affecting food choice and social
seasonal frugivores: Use of variable resources, Am J Primatol organization in Pan paniscus, PhD dissertation, New York, 1990,
43:87–109, 1997. Stony Brook University, NY.
2. Remis MJ, Dierenfeld ES: Digesta passage, digestibility and 22. Hohmann G, Fruth B: New records on prey capture and meat
behavior in captive gorillas under two dietary regimens, Int J eating by bonobos at Lui Kotale, Salonga National Park, Demo-
Primatol 25(4):825–845, 2004. cratic Republic of Congo, Folia Primatol (Basel) 79:103–110,
3. Rogers ME, Abernethy K, Bermejo M, et al: Western gorilla 2007.
diet: A synthesis from six sites, Am J Primatol 64(2):173–192, 23. McGrew WC, Marchant LF, Beuerlein MM, et al: Prospects for
2004. bonobo insectivory: Lui Kotal, Democratic Republic of Congo,
4. Bastain ML, Zweifel N, Vogel ER, et al: Diet traditions in wild Int J Primatol 28(6):1237–1252, 2007.
orangutans, Am J Phys Anthropol 143:175–187, 2010. 24. Sakamaki T, Maloueki U, Bakaa B, et al: Mammals consumed by
5. Hamilton RA, Galdikas BMF: A preliminary study of food bonobos (Pan paniscus): new data from Iyondji forest, Tshuapa,
selection by the orangutan in relation to plant quality, Primates Democratic Republic of Congo, Primates 57:295–301, 2016.
35:255–263, 1994. 25. Oelze VM, Fuller BT, Richards MP, et al: Exploring the contri-
6. Knott CD: Changes in orangutan caloric intake, energy balance, bution and significance of animal protein in the diet of bonobos
and ketones in response to fluctuating fruit availability, Int J by stable isotope analysis of hair, PNAS 108(24):9792–9799,
Primatol 19:1061–1079, 1998. 2011.
7. Knott CD: Reproductive, physiological and behavioral responses 26. Utami SS, van Hooff JA: Meat-eating by adult female Sumatran
of orangutans in Borneo to fluctuations in food availability. PhD orangutans (Pongo pygmaeus abelii), Am J Primatol 43:159–165,
dissertation, Cambridge, MA, 1999, Harvard University. 1997.
CHAPTER 83 Great Ape Nutrition 595
27. van Schaik CP, van Noordwijk MA, Vogel ER: Ecological sex lowland gorilla (Gorilla gorilla) and a Sumatran orangutan (Pongo
differences in wild orangutans. In Wich SA, Utami Atmoko abelii), Am J Primatol 79e22609, 2016.
SS, Setia T, et al, editors: Orangutans: geographic variation in 41. Popovich DG, Jenkins DJ, Kendall CW, et al: The western
behavioral ecology and conservation, New York, 2009, Oxford lowland gorilla diet has implications for the health of humans
University Press, pp 49–64. and other hominoids, J Nutr 127:2000–2005, 1997.
28. Hardus ME, Lameira AR, Zulfa A, et al: Behavioral, ecological, 42. Avena NM, Rada P, Hoebel BG: Evidence for sugar addiction:
and evolutionary aspects of meat-eating by Sumatran orangutan Behavioral and neurochemical effects of intermittent, excessive
(Pongo abelii), Int J Primatol 33:287–304, 2012. sugar intake, Neurosci Biobehav Rev 32:20–39, 2008.
29. Silva M, Downing JA: CRC handbook of mammalian body masses, 43. National Research Council: Nutrient requirements of nonhuman
Boca Raton, FL, 1995, CRC Press. primates, ed 2, Washington, DC, 2003, National Academies
30. Coolidge HJ, Shea BT: External body dimensions of Pan paniscus Press.
and Pan troglodytes chimpanzees, Primates 23:245–251, 1982. 44. Strong V, Grindlay D, Redrobe S, et al: A systematic review of
31. Markham R, Groves CP: Weights of wild orangutans, Am J Phys the literature relating to captive great ape morbidity and mortal-
Anthropol 81:1–3, 1990. ity, J Zoo Wildl Med 47(3):697–710, 2016.
32. Grzimk B: Masse und gewichtevon flachland-gorillas, Z Sauget- 45. Vlčová K, Gomez A, Petrželková K, et al: Effect of antibiotic
ierkd 21:192–194, 1956. treatment on the gastrointestinal microbiome of free-ranging
33. Rothman JM, Dierenfeld ES, Hintz HF, et al: Nutritional quality Western Lowland Gorillas (Gorilla g. gorilla), Microb Ecol 72:
of gorilla diets: consequences of age, sex, and season, Oecologia 943–954, 2016.
155:111–122, 2008. 46. Hensrud D: Obesity. In Rakel RE, Bope ET, editors: Conn’s
34. Cassella CM, Mills A, Lukas KE: Prevalence of regurgitation and current therapy, Philadelphia, PA, 2002, Saunders, pp 577–584.
reingestion in orangutans housed in North American zoos and 47. Klenov VE, Jungheim ES: Obesity and reproductive function: a
an examination of factors influencing its occurrence in a single review of the evidence, Curr Opin Obstet Gynecol 26:455–460,
group of Bornean orangutans, Zoo Biol 609–620, 2012. 2014.
35. Pretorius IS: Utilization of polysaccharides by Saccharomyces 48. Plowman A: Fruit-free diets for primates, in Proceedings of the
cerevisiae. In Zimmermann FK, Entian KD, editors: Yeast sugar 11th Conference on Zoo and Wildlife Nutrition 1–4, 2015.
metabolism, bichemistry, genetics, biotechnology, and applications, 49. Johnson AW, Galagher M: Greater effort boosts the affective
Lancaster, PA, 1997, Technomic Publishing Company. properties of food, Proc Biol Sci 278:1450–1456, 2011.
36. Junge RE, Gannon FH, Porton I, et al: Management and preven- 50. Schulman FY, Farb A, Virmani R, et al: Fibrosing cardiomyopa-
tion of vitamin D deficiency rickets in captive-born juvenile thy in captive western lowland gorillas (Gorilla gorilla gorilla)
chimpanzees (Pan troglodytes), J Zoo Wildl Med 31:361–369, in the United States: a retrospective study, J Zoo Wildl Med
2000. 26:43–51, 1995.
37. Meehan TP: Nutritional secondary hyperparathyroidism in an 51. Varki N, Anderson D, Herndon JG, et al: Heart disease is
infant gorilla (Gorilla gorilla). In Proceedings of the American common in humans and chimpanzees, but is caused by different
Association of Zoo Veterinarians 126, 1993. pathological processes, Evol Appl 2:101–112, 2009.
38. Holick MF, Chen TC: Vitamin D deficiency: a worldwide problem 52. McManamon R, Lowenstine L: Cardiovascular disease in great
with health consequences, Am J Clin Nutr 87:1080S–1086S, apes. In Miller RE, Fowler ME, editors: Fowler’s zoo and wild
2008. animal medicine current therapy (vol 7). St. Louis, 2011, Elsevier,
39. Lukas KE: A review of nutritional and motivational factors con- pp 408–415.
tributing to the performance of regurgitation and reingestion in 53. Great Ape Heart Project white paper: The great ape heart project
captive lowland gorillas (Gorilla gorilla gorilla), Appl Anim Behav (GAHP): a collaboration to understand heart disease, reduce
Sci 63:237–249, 1999. mortality and improve cardiac health in all four great ape taxa.
40. Power ML, Schulkin J, Drought H, et al: Patterns of milk mac- [http://greatapeheartproject.files.wordpress.com/2011/11/
ronutrients and bioactive molecules across lactation in a western gahp_whitepaper2012.pdf ], 2012.
SECTION 17
Marine Mammals
84 Marine Mammal Viruses, 597
596
84
Marine Mammal Viruses
JIM WELLEHAN AND GALAXIA CORTES-HINOJOSA
V
iral abundance in the ocean has been estimated a result, they may change much more rapidly, and this
at 1030 virions, one or two orders of magnitude is why a segmented influenza virus changes to the extent
greater than the estimated number of bacterial and where novel vaccines are needed every year or two, whereas
eukaryotic cells, so viruses are the most abundant life in vaccines for an otherwise biologically similar nonsegmented
the oceans.1,2 Marine mammals live in intimate contact morbillivirus result in lifelong immunity, because the virus
with the ocean, and therefore a vast diversity of viruses. cannot change as rapidly.
Advances in molecular techniques have allowed a rapid Presence of an envelope is also clinically relevant. Viruses
expansion of knowledge on viral diversity, enabling greater with envelopes tend to need them to infect target cells. This
understanding of evolution, causes of mortalities, and lipid layer is more susceptible to environmental conditions
pathogen interactions. and disinfectants, consequently making cleaning easier and
decreasing environmental persistence.
Clinical Implications of Viral Biology
Viruses are classified on the basis of their genomic mate- RNA Viruses
rial and key elements of structure and replication. A basic Astroviridae
knowledge of these elements is also clinically useful for
predicting disease risk and epidemiologic characteristics, Astroviruses are small (28–30 nm), spherical, nonenveloped,
and for establishing appropriate management protocols positive-sense, single-stranded viruses with intracytoplasmic
(Table 84.1). DNA viruses often replicate in the nucleus replication. Astroviruses often cause diarrhea and have a
of the host cell, and tend to be more species specific. RNA high prevalence in children; most children over 5 years of age
viruses tend to replicate in the cytoplasm and are more have antibodies to human astroviruses. There are also several
prone to jumping between species. There are exceptions, astroviruses documented in association with encephalitis in
such as poxviruses, asfarviruses, and iridoviruses, which nonmarine mammals. In the order Carnivora, astroviruses
are DNA viruses that replicate in the cytoplasm and have have been associated with diarrhea in mink (Neovison vison),
significantly lower host fidelity, and bornaviruses, which are domestic dogs, cheetahs (Acinonyx jubatus), and domestic
RNA viruses with intranuclear replication. cats.3 In marine mammals, astroviruses have been reported
Viral genome size is also important. Large viruses with in bottlenose dolphins (Tursiops truncatus),4 Steller sea lions
many genes are better at evolving complex host interactions (Eumetopias jubatus),4 minke whales (Balaenoptera acuto-
for things like latency. Smaller viruses are capable of more rostrata), and California sea lions (Zalophus californianus).4,5
rapid evolution and therefore more capable of adapting to Five different astroviruses have been found: one in bottlenose
different hosts and tissues, resulting in host jumping and dolphins, one in Steller sea lions, and three in California
differing clinical presentations. sea lions.4 Next-generation sequencing (NGS) has identified
Genome organization is also clinically relevant; nonseg- a total of eight additional astroviruses in California sea
mented viruses that are not very capable of recombination, lions.5 This study was done using fecal swabs of animals in
such as paramyxoviruses, largely change through muta- rehabilitation facilities and found an astrovirus prevalence
tion. Mutations are much more likely to be deleterious of 51%.5 Bottlenose Dolphin Astrovirus 1 (BDAstV1) was
rather than beneficial, limiting rates of change. Segmented found in 86% of stranded bottlenose dolphins and in 50%
viruses, such as orthomyxoviruses or reoviruses, may swap of healthy bottlenose dolphins from a managed collection.
segments, much like sexual reproduction in a eukaryote. A subset of dolphins who were persistent shedders were
This enables an organism to use homologous genes that identified. The clinical significance of astroviruses in marine
have been functional in another conspecific, which is much mammals is likely to be greatest in young animals as a cause
more likely to be beneficial than random mutations. As of diarrhea, as is seen in other species.
597
598 SE C T I O N 17 Marine Mammals
TABLE Virus Taxa Associated With Clinical the captive harbor seals, the clinical signs included anorexia
84.1 Presentations in Marine Mammals with abnormal behavior after the sudden death of two other
harbor seals in the same pool. At clinical examination, the
Clinical Signs Differentials mucosa was purple and injected, and abnormal pulmo-
Mucocutaneous Poxvirus, herpesvirus, calicivirus, nary sounds were auscultated; blood work abnormalities
lesions papillomavirus, picornavirus included leukocytosis, hypernatremia, and hyperchloremia.
Skin Poxvirus, herpesvirus, calicivirus, Postmortem findings included acute necrotizing enteritis
papillomavirus and pulmonary edema. Confirmation of coronavirus was
attempted on cell culture but was not possible at the time.
Respiratory Influenza, morbillivirus, calicivirus,
coronavirus, adenovirus In the epizootic event in wild harbor seals, the five carcasses
in suitable condition had lymphocytic/histocytic necrotizing
Enteritis Adenovirus, astrovirus, coronavirus
pneumonia diagnosed histologically. An Alphacoronavirus
Neurologic Morbillivirus, herpesvirus was diagnosed in lung tissue using consensus polymerase
Neoplasia Herpesvirus, retrovirus, chain reaction (PCR) with product sequence identification.
polyomavirus, papillomavirus
Hepatitis Adenovirus, coronavirus, Orthomyxoviridae (Influenza Virus)
herpesvirus
Orthomyxoviruses are medium sized (80–120 nm), seg-
Ocular Adenovirus, herpesvirus, calicivirus
mented, pleomorphic, enveloped, negative-sense, single-
stranded RNA viruses with intranuclear and intracytoplasmic
replication. Influenza viruses are further divided into the
genera Influenzavirus A, Influenzavirus B, and Influenzavirus
Caliciviridae C. Influenza A viruses tend to be the most pathogenic
and have been well known to cause pandemic outbreaks
San Miguel sea lion virus (SMSV) is a small (30–38 nm) in humans since 1918/1919. In marine mammals, both
icosahedral, nonenveloped, positive-sense, single-stranded Influenza A and Influenza B have been diagnosed in wild
virus with intracytoplasmic replication that belongs to the populations of cetaceans and pinnipeds since the late 1970s.
genus Vesivirus in the family Caliciviridae. This virus is Mass mortalities in association with several different strains
genetically indistinguishable from vesicular exanthema of of Influenza A have been reported in harbor seals since 1979.
swine virus (VESV), which is a reportable foreign animal Influenza B has been reported in stranded harbor seals since
disease that has officially been considered eradicated in 1999, but not in association with epidemics. In cetaceans,
the United States since 1956. Perhaps most concerning, Influenza A has been detected in stranded long-finned pilot
related vesiviruses have been found to rapidly evolve greater whales. Additionally, South American fur seals (Arctocepha-
virulence where there are high host population densities.6 lus australis),16 Caspian seals (Pusa caspica),17 Baikal seals
Serologic studies suggest exposure to vesiviruses in cetaceans (Phoca sibirica) and ringed seals (Phoca hispida),18 Dall’s
and pinnipeds.7 In pinnipeds, SMSV clinical manifesta- porpoises (Phocoenoides dalli),19 beluga whales, and other
tions include vesicular lesions on the flippers and the species have been reported to be seropositive for influenza.20
mouth, as well as gastroenteritis in California sea lions.8 Clinical signs include epistaxis, conjunctivitis, subcutaneous
Clinical presentations in premature pups include respira- emphysema, and general weakness. Diagnosis of influenza
tory distress and locomotor impairment, with possible fatal can be based on a combination of clinical signs, qPCR,
consequences. Cutaneous lesions typically resolve without serology, and postmortem findings (gross pathology, his-
supportive care. SMSV has zoonotic potential, causing an topathology, and immunohistochemistry). Seal-to-human
influenza-like syndrome followed by blisters on the hands influenza transmission has been described. Direct contact
and feet.9 SMSV has also been associated with hepatitis in on massive mortalities and one case of direct contact with
humans.10,11 respiratory secretion are associated with conjunctivitis in
humans.21 A comprehensive review of influenza in marine
Coronaviridae mammals is available.20
Paramyxoviridae
Coronaviruses are large (120–160 nm), round, toroidal
or bacilliform, enveloped, positive-sense, single-stranded Morbillivirus
viruses with intracytoplasmic replication. In marine Paramyxoviruses are pleomorphic, enveloped, negative-
mammals, viruses in the genus Gammacoronavirus have sense, single-stranded viruses with intracytoplasmic
been characterized in a captive beluga whale (Delphinapterus replication. This family causes respiratory, neurologic, and
leucas)12 and bottlenose dolphins,13 and an Alphacoronavirus multisystemic diseases in mammals.22 In marine mammals,
has been found in both captive and wild harbor seals (Phoca three viruses in the genus Morbillivirus have been described:
vitulina).14,15 Clinical signs in the beluga whale included canine distemper virus (CDV), phocine distemper virus
generalized pulmonary disease and acute liver failure. For (PDV), and cetacean morbillivirus (CeMV). CeMV has
CHAPTER 84 Marine Mammal Viruses 599
significant diversity and may eventually be split into signs and use of PCR diagnostics and/or negative stain-
more than one species; morbilliviruses from the North- ing electron microscopy of feces from clinical cases, with
ern Hemisphere [porpoise morbillivirus (PMV), dolphin intranuclear inclusions present on histologic examination in
morbillivirus (DMV), pilot whale morbillivirus (PWMV), postmortem cases.
and Longman’s beaked whale morbillivirus (BWMV)] show
significant divergence from CeMV from the Southern Herpesviridae
Hemisphere (from Swan River, Australia and from Brazil).
Morbilliviruses have been recognized as causes of mass Herpesviruses are large (100–150 nm), icosahedral, envel-
mortalities in pinnipeds and cetaceans since the early 1980s. oped, double-stranded viruses with intranuclear replication.
Diagnosis in cetaceans is usually postmortem, but clinical Herpesviruses can be latent in different sites, with certain
signs include cachexia, abnormal mentation, and respira- subfamily tendencies; alphaherpesviruses often establish
tory distress. In pinnipeds, clinical signs include respiratory latency in neurons, whereas betaherpesviruses and gamma-
distress, ocular and nasal discharge, pyrexia, and erratic herpesviruses establish latency in leukocytes. Herpesviruses
swimming. Morbilliviruses infect via the signaling lym- infect a wide range of vertebrates. Trichechid herpesvirus 1
phocytic activation molecule (SLAM [CD150]) receptor, (TrHV-1) is a gammaherpesvirus reported in West Indian
taking out memory T cells and leaving the host significantly manatees (Trichechus manatus) and may be a biomarker for
immunosuppressed. Coinfection with other pathogens such stress.36
as Aspergillus fumigatus or Brucella become more clinically Polar bears (Ursus maritimus) are susceptible to enceph-
significant.23,24 Comprehensive reviews are available for alitis caused by the alphaherpesvirus equine herpes virus
both pinnipeds22 and cetaceans.23 9 (EHV-9), which may be fatal (see Chapter 33).37 This
virus appears to be endemic in Grevy’s zebras (Equus
Respirovirus grevyi) and is associated with fatalities in diverse laura-
The genus Respirovirus contains parainfluenza viruses siatherian mammals. Although it is unlikely to be signifi-
infecting several mammal hosts. Parainfluenza virus was cant in wild populations due to low contact rates with the
first identified in marine mammals in a bottlenose dolphin endemic host, it is a more significant concern in zoological
kept in an open water enclosure in 2008. Clinical signs collections.
included respiratory stridor, halitosis, and exudate from the In pinnipeds, 11 herpesviruses have been reported to
blowhole.25 Serologic data suggested that exposure may be date.38,39 Otarine herpesvirus 1 (OtHV-1) is a gammaher-
common in wild and captive bottlenose dolphins.26 This pesvirus associated with urogenital carcinoma in California
virus is closely related to Bovine respirovirus virus 3 and sea lions. OtHV-1 was also detected in a captive South
Human respirovirus virus 3, and potential risks for host American fur seal (Arctocephalus australis) with urogenital
jumping of these viruses should be considered. carcinoma.40 This virus has a high prevalence in wild adult
California sea lions (46% in males and 22% in females).
Rates of metastasis are high, with sublumbar lymph nodes
DNA Viruses and liver as common sites.41,42 Otarine herpesvirus 4, found
Adenoviridae in clinically unaffected northern fur seals (Callorhinus
ursinus), is a very close relative of OtHV-1; investigation of
Adenoviruses are medium-sized (70–90 nm) icosahedral, this virus as a potential vaccine against OtHV-1 is merited.
nonenveloped, double-stranded DNA viruses with Phocid herpesvirus 1 (PHV-1) is an alphaherpesvirus
intranuclear replication. In marine mammals, adeno- seen in fatal outbreaks in harbor seals and gray seals (Hali-
viruses have been reported in pinnipeds, mustelids, and choerus grypus) in rehabilitation facilities, affecting mainly
cetaceans. Adenoviruses have been associated with death young and immunosuppressed individuals. This virus may
in yearling California sea lions in rehabilitation; clinical target the respiratory system as well as the GI tract, adrenal
signs included photophobia, emaciation, blood-tinged diar- glands, and liver. Clinical signs may include nasal and ocular
rhea, leukopenia, and monocytosis.27 In 2011 California discharge, coughing, inflammation of the oral mucosa,
sea lion adenovirus 1 (CSLAdV-1) was characterized as a vomiting, diarrhea, lethargy, anorexia, and fever.43–45
cause of viral hepatitis in wild animals and has since been In cetaceans, diverse alpha- and gammaherpesviruses
detected in several pinniped species in captivity, especially have been reported. Delphinid herpesviruses 1 and 2, both
in elder animals.28–30 California sea lion adenovirus 2 and alphaherpesviruses, have been associated with fatal necrosis
phocid adenoviruses 1 and 2 have been identified in ocular of the spleen, thymus, and lymph nodes in bottlenose
lesions and in unaffected eyes of pinnipeds.31 In cetaceans, dolphins, with intranuclear inclusions present in affected
adenoviruses have been associated with gastrointestinal tissues.46 A harbor porpoise herpesviral encephalitis case was
(GI) symptoms in bottlenose dolphins and harbor por- also likely to have been caused by an alphaherpesvirus. Del-
poises from Europe32,33 and the United States.34 Recently, phinid herpesvirus 4, a gammaherpesvirus highly prevalent
a novel adenovirus with unknown clinical significance in bottlenose dolphins, is associated with plaque-like genital
was found in Southern sea otters.35 Diagnosis of adeno- lesions, and gammaherpesviruses have been associated with
viral disease may be made with a combination of clinical similar lesions in other cetacean species.38,47,48
600 SE C T I O N 17 Marine Mammals
commonly called qPCR or real-time PCR, such as SYBR 14. Nollens HH, Wellehan JF, Archer L, et al: Detection of a respi-
Green, which involves dye incorporation of dye into any ratory coronavirus from tissues archived during a pneumonia
amplified DNA and is less rigorous. epizootic in free-ranging Pacific harbor seals Phoca vitulina
richardsii, Dis Aquat Organ 90:113–120, 2010.
PCR-based methods are limited because they require
15. Bossart G, Schwartz J: Acute necrotizing enteritis associated with
a known conserved area of sequence for primer design. suspected coronavirus infection in 3 harbor seals (Phoca vitulina),
The detection of novel phylogenetically distant viruses may J Zoo Wildl Med 21:84–87, 1990.
be challenging or even impossible. In the last few years, 16. Blanc A, Ruchansky D, Clara M, et al: Serologic evidence of
NGS technologies have developed. Rather than targeted influenza A and B viruses in South American fur seals (Arcto-
sequencing of a single DNA template, NGS methods may cephalus australis), J Wildl Dis 45:519–521, 2009.
sequence literally millions of different nucleic acid templates 17. Ohishi K, Ninomiya A, Kida H, et al: Serological evidence of
in a sample. Although NGS can obtain sequences from transmission of human influenza A and B viruses to Caspian
previously unknown and divergent pathogens, it must also seals (Phoca caspica), Microbiol Immunol 46:639–644, 2002.
recognize and sift through literally millions of nontarget 18. Ohishi K, Kishida N, Ninomiya A, et al: Antibodies to human-
sequences. This is a major bioinformatics challenge but related H3 influenza A virus in Baikal seals (Phoca sibirica)
and ringed seals (Phoca hispida) in Russia, Microbiol Immunol
has already resulted in significant advances in infectious
48:905–909, 2004.
disease. Costs for NGS and the time required for data 19. Ohishi K, Maruyama T, Ninomiya A, et al: Serologic inves-
analysis are still high but have dramatically decreased, and tigation of influenza A virus infection in cetaceans from the
NGS technologies are already in use for marine mammal western North Pacific and the Southern oceans, Mar Mam Sci
virus discovery. 22:214–221, 2006.
20. Fereidouni S, Munoz O, Von Dobschuetz S, et al: Influenza virus
References infection of marine mammals, Ecohealth 13:161–170, 2016.
21. Webster RG, Geraci J, Petursson G, et al: Conjunctivitis in
1. Suttle CA: Marine viruses—major players in the global ecosys- human beings caused by influenza A virus of seals, N Engl J
tem, Nature Rev Microbio 5:801–812, 2007. Med 304:911, 1981.
2. Suttle CA: Viruses in the sea, Nature 437:356–361, 2005. 22. Wang L, Collins P, Fouchier R, et al: Family paramyxoviridae.
3. Atkins A, Wellehan J, Childress A, et al: Characterization of an Virus taxonomy: Ninth Report of the International Committee
outbreak of astroviral diarrhea in a group of cheetahs (Acinonyx on Taxonomy of Viruses 2012:672–685.
jubatus), Vet Microbiol 136:160–165, 2009. 23. Cassle SE, Landrau-Giovannetti N, Farina LL, et al: Coinfection
4. Rivera R, Nollens H, Venn-Watson S, et al: Characterization of by Cetacean morbillivirus and Aspergillus fumigatus in a juvenile
phylogenetically diverse astroviruses of marine mammals, J Gen bottlenose dolphin (Tursiops truncatus) in the Gulf of Mexico, J
Virol 91:166–173, 2010. Vet Diagn Invest 28:729–734, 2016.
5. Li L, Shan T, Wang C, et al: The fecal viral flora of California 24. West KL, Levine G, Jacob J, et al: Coinfection and vertical
sea lions, J Virol 85:9909–9917, 2011. transmission of Brucella and Morbillivirus in a neonatal sperm
6. Schorr-Evans E, Poland A, Johnson W, et al: An epizootic of whale (Physeter macrocephalus) in Hawaii, USA, J Wildl Dis
highly virulent feline calicivirus disease in a hospital setting in 51:227–232, 2015.
New England, J Feline Med Surg 5:217–226, 2003. 25. Nollens HH, Wellehan JFX, Saliki JT, et al: Characterization of
7. Akers T, Smith A, Latham A, et al: Calicivirus antibodies in a parainfluenza virus isolated from a bottlenose dolphin (Tursiops
California gray whales (Eschrichtius robustus) and Steller sea lions truncatus), Vet Microbiol 128:231–242, 2008.
(Eumetopias jupatus), Arch Virol 46:175–177, 1974. 26. Venn-Watson S, Rivera R, Smith CR, et al: Exposure to novel
8. Schmitt TL, Reidarson TH, St. Leger J, et al Novel presenta- parainfluenza virus and clinical relevance in 2 bottlenose dolphin
tion of San Miguel sea lion virus (Calicivirus) epizootic in adult (Tursiops truncatus) populations, Emerg Infect Dis 14:397–405,
captive sea lions, Zalophus californianus. In Proceedings of the 2008.
40th International Association for Aquatic Animal Medicine, 27. Dierauf L, Lowenstine L, Jerome C: Viral hepatitis (adenovirus)
2009. in a California sea lion, J Am Vet Med Assoc 179:1194–1197,
9. Smith AW, Skilling DE, Cherry N, et al: Calicivirus emergence 1981.
from ocean reservoirs: zoonotic and interspecies movements, 28. Inoshima Y, Murakami T, Ishiguro N, et al: An outbreak of
Emerg Infect Dis 4:13–20, 1998. lethal adenovirus infection among different otariid species, Vet
10. Smith AW, Iversen PL, Skilling DE, et al: Vesivirus viremia and Microbiol 165:455–459, 2013.
seroprevalence in humans, J Med Virol 78:693–701, 2006. 29. Goldstein T, Colegrove K, Hanson M, et al: Isolation of a novel
11. Lee H, Cho YH, Park JS, et al: Elevated post-transfusion serum adenovirus from California sea lions (Zalophus californianus), Dis
transaminase values associated with a highly significant trend Aquat Organ 94:243–248, 2011.
for increasing prevalence of anti-Vesivirus antibody in Korean 30. Cortés-Hinojosa G, Doescher B, Kinsel M, et al: Coinfection
patients, J Med Virol 84:1943–1952, 2012. of California sea lion adenovirus 1 and a novel polyomavirus in
12. Mihindukulasuriya KA, Wu G, St Leger J, et al: Identification Hawaiian monk seal (Neomonachus schauinslandi), J Zoo Wildl
of a novel coronavirus from a beluga whale by using a panviral Med 47:427–437, 2016.
microarray, J Virol 82:5084–5088, 2008. 31. Wright EP, Waugh LF, Goldstein T, et al: Evaluation of viruses
13. Woo PC, Lau SK, Lam CS, et al: Discovery of a novel bottlenose and their association with ocular lesions in pinnipeds in rehabili-
dolphin coronavirus reveals a distinct species of marine mammal tation, Vet Ophthalmol 18(Suppl 1):148–159, 2015.
coronavirus in Gammacoronavirus, J Virol 88:1318–1331, 32. Rubio-Guerri C, García-Párraga D, Nieto-Pelegrín E, et al: Novel
2014. adenovirus detected in captive bottlenose dolphins (Tursiops
602 SE C T I O N 17 Marine Mammals
truncatus) suffering from self-limiting gastroenteritis, BMC Vet 44. Gulland F, Lowenstine L, Lapointe J, et al: Herpesvirus infection
Res 11:53, 2015. in stranded Pacific harbor seals of coastal California, J Wildl Dis
33. van Beurden SJ, IJsseldijk LL, van de Bildt MW, et al: A novel 33:450–458, 1997.
cetacean adenovirus in stranded harbour porpoises from the 45. Harder T, Vos H, deSwart R, et al: Age-related disease in recur-
North Sea: detection and molecular characterization, Arch Virol rent outbreaks of phocid herpesvirus type-1 infections in a seal
162:2035–2040, 2017. rehabilitation centre: Evaluation of diagnostic methods, Vet Rec
34. Standorf K, Cortés-Hinojosa G, Venn-Watson S, et al: Phyloge- 140:500–503, 1997.
netic analysis of the genome of an enteritis-associated bottlenose 46. Blanchard T, Santiago N, Lipscomb T, et al: Two novel alpha-
dolphin mastadenovirus supports a clade infecting the Cetartio- herpesviruses associated with fatal disseminated infections in
dactyla, J Wildl Dis 54:112–121, 2018. Atlantic bottlenose dolphins, J Wildl Dis 37:297–305, 2001.
35. Siqueira JD, Ng T, Miller M, et al: Endemic infection of stranded 47. van Beurden SJ, IJsseldijk LL, Ordonez SR, et al: Identification
Southern Sea Otters (Enhydra lutris nereis) with novel parvovirus, of a novel gammaherpesvirus associated with (muco)cutaneous
polyomavirus, and adenovirus, J Wildl Dis 2017. lesions in harbour porpoises (Phocoena phocoena), Arch Virol
36. Ferrante JA, Cortés-Hinojosa G, Archer LL, et al: Development 160:3115–3120, 2015.
of a quantitative PCR assay for measurement of Trichechid 48. van Elk C, van de Bildt M, de Jong A, et al: Genital herpesvirus
herpesvirus 1 (TrHV1) load in the Florida manatee (Trichechus in bottlenose dolphins (Tursiops truncatus): cultivation, epidemi-
manatus latirostris), J Vet Diagn Invest 29:476–482, 2017. ology, and associated pathology, J Wildl Dis 45:895–906, 2009.
37. Donovan T, Schrenzel M, Tucker T, et al: Meningoencephalitis 49. Ghim S-J, Joh J, Mignucci-Giannoni AA, et al: Genital papillo-
in a polar bear caused by equine herpesvirus 9 (EHV-9), Vet matosis associated with two novel mucosotropic papillomaviruses
Pathol 46:1138–1143, 2009. from a Florida manatee (Trichechus manatus latirostris), Aquatic
38. Maness H, Nollens H, Jensen E, et al: Phylogenetic analysis Mammals 40:195–200, 2014.
of marine mammal herpesviruses, Vet Microbiol 149:23–29, 50. Robles-Sikisaka R, Rivera R, Nollens H, et al: Evidence of
2011. recombination and positive selection in cetacean papillomavi-
39. Cortés-Hinojosa G, Gulland FM, DeLong R, et al: A novel ruses, Virology 427:189–197, 2012.
gammaherpesvirus in Northern fur seals (Callorhinus ursinus) 51. Rivera R, Robles-Sikisaka R, Hoffman E, et al: Characterization
is closely related to the California sea lion (Zalophus califor- of a novel papillomavirus species (ZcPV1) from two California
nianus) carcinoma-associated otarine herpesvirus-1, J Wildl Dis sea lions (Zalophus californianus), Vet Microbiol 155:257–266,
52:88–95, 2016. 2012.
40. Dagleish M, Barrows M, Maley M, et al: The first report of 52. Bracht A, Brudek R, Ewing R, et al: Genetic identification
otarine herpesvirus-1-associated urogenital carcinoma in a of novel poxviruses of cetaceans and pinnipeds, Arch Virol
South American fur seal (Arctocephalus australis), J Comp Pathol 151:423–438, 2006.
149:119–125, 2013. 53. Burek K, Beckmen K, Gelatt T, et al: Poxvirus infection of Steller
41. Buckles E, Lowenstine L, DeLong R, et al: Age-prevalence of sea lions (Eumetopias jubatus) in Alaska, J Wildl Dis 41:745–752,
otarine herpesvirus-1, a tumor-associated virus, and possibility 2005.
of its sexual transmission in California sea lions, Vet Microbiol 54. Tuomi PA, Murray MJ, Garner MM, et al: Novel poxvirus infec-
120:1–8, 2007. tion in northern and southern sea otters (Enhydra lutris kenyoni)
42. Lipscomb T, Scott D, Garber R, et al: Common metastatic car- and Enhydra lutris neiris), Alaska and California, USA, J Wildl
cinoma of California sea lions (Zalophus californianus): Evidence Dis 50:607–615, 2014.
of genital origin and association with novel gammaherpesvirus, 55. Nollens H, Ruiz C, Walsh M, et al: Cross-reactivity between
Vet Pathology 37:609–617, 2000. immunoglobulin G antibodies of whales and dolphins correlates
43. Gulland FM: Stranded seals: important sentinels, J Am Vet Med with evolutionary distance, Clin Vaccine Immunol 15:1547–1554,
Assoc 214:1191–1192, 1999. 2008.
85
Mycobacterium pinnipedii
ALEXIS LÉCU
T
he Mycobacterium tuberculosis complex (MTBC) The isolates are usually susceptible to isoniazid, rifampicin,
stands as a group of mycobacteria species of major streptomycin, ethambutol, and para-aminosalicylic acid.
concern, mostly because of their high zoonotic Colonies are dysgonic, rough, and nonchromogenic.11
abilities. The constituent members of the MTBC are However, biochemical and cultural characteristics do not
highly genetically related (0.03%) and can be split broadly clearly differentiate M. pinnipedii from M. bovis: genomic
between human and animal-adapted strains: the major features are most often the key to differentiating M. pin-
human pathogens, where no obvious animal reservoir has nipedii from all other MTBC species.
been identified, are Mycobacterium tuberculosis and M. All MTBC species share a similar 16S RNA gene
africanum (subtypes 1 and 2); the animal-adapted strains sequence except M. pinnipedii, which has one single dif-
have been isolated from a range of wild and domesticated ferent nucleotide within this sequence. Phylogenetically,
animals and are named after their host of initial/most these species share the same GyrB sequence motif with
frequent isolation,1 including M. bovis in ruminants, M. M. africanum type I and M. canetti.12 The genomic profile
microti in rodents, cats, and south American camelids, of MTBC species is very different from M. bovis and M.
M. caprae in goats, M. orygis in oryx and other antelopes, caprae, which underwent more deletions, but it has the
M. mungi in banded mongoose (Mungos), the Dassie same deleted regions (namely RD7, RD8, and RD10) as
bacillus in rock hyrax (Procavia) and M. pinnipedii (M. do M. microti and both M. orygis and the Dassie bacillus.
p) in seals and sea lions. Actually, recent phylogenic The deletions that are specific to M. pinnipedii species are
studies2,3 are establishing an evolution route leading M. the sequences RD2seal and RDpin: the presence of these
pinnipedii from original African pinniped host species to deletions is systematic in all strains and is independent from
South American marine mammals through the ocean, geographic origin or host type. It should also be noted that
and then to an adaptation to human beings in the South the RD2 sequence has been demonstrated as essential to the
American continent, leading to a lineage of M. tuberculosis full virulence of MTBC,13 so RD2seal deletion or mutation
adapted to Homo sapiens more than 1000 years ago on this could lead to a reduction of pathology extension or organ
continent. burden in hosts, as for the Bacillus Calmette-Guérin (BCG)
strain.14 This could be linked to the suspected “latency”
Etiology and Hosts feature in some of the host species.
All M. pinnipedii isolates contain the sequences IS6110,
M. pinnipedii was formerly known as “seal bacillus” as it IS1081, MPB70, MPB83, and MTP40 but fail to produce
was first found in a colony of three different species of detectable MPB70 and MPB83 antigens15; therefore, use
otariid seals in Western Australia between 1981 and 1986.4 of serodiagnostic methods based on MPB70 and MPB83
There were several papers between 1990 and 2000 where antibody detection may be of questionable help in M. pin-
“tuberculosis outbreaks” in pinnipeds and other species were nipedii screening, and the lack of these antigen expressions
reported,5–8 occasionally wrongly classified as M. africanum may even be used as a key in distinguishing M. pinnipedii
or M. bovis infections, and retrospectively attributable to M. infection from other MTBC species.16
pinnipedii. This mycobacterium was definitively classified as Genotyping, especially spoligotyping, is still the most effi-
a standalone species of the MTBC in 2003, mainly based cient way to differentiate M. pinnipedii from other MTBC
on its molecular9 and genomic characteristics.10 species. The spoligotyping method is based on polymerase
Regarding phenotype, M. pinnipedii is a slow-growing chain reaction (PCR) amplification of a highly polymorphic
mycobacterium, which means that on primary isolation, direct repeat locus in the MTBC genome. Results can be
there is minimal visible growth during the first week of obtained from a pure culture or also from a raw sample
culture. Primary culture is obtained at 37°C on egg-based containing enough M. pinnipedii DNA, if obtained within
medium in 3–6 weeks on average, and antibiotic suscepti- one day. All isolated M. pinnipedii spoligotype patterns
bility could be determined after an additional 2–4 weeks. form a cluster that is clearly different from those of all
603
604 SE C T I O N 17 Marine Mammals
other members of the MTBC.17 Thus, the clinical useful- same routes; for example, aerosols created by high-pressure
ness of spoligotyping is determined by its rapidity, both in cleaning of pinniped enclosures were found to create a
detecting causative bacteria and in providing epidemiologic contaminating fog in distant tapir or camel enclosures,19,20
information on strain identities. and close contact can be problematic, for example, between
a pinniped and its human trainer.7,21
Hosts Contamination from pinnipeds to terrestrial mammals
also occurred in the wild, with cattle infected while grazing
M. pinnipedii is historically known as the “seal bacillus” and on areas adjacent to seal beaches.22,23 At necropsy, lesions in
was initially discovered only in species from the Otariidae thoracic lymph nodes suggest aerosol transmission in those
family, but was eventually determined to be able to infect “non-zoo” species.
other marine and terrestrial mammals, either naturally or
experimentally (Box 85.1). Postmortem Diagnosis
“Natural” wildlife hosts all originate from the southern
hemisphere and outbreaks in wild pinniped colonies are For every suspected tuberculosis case, and for any dead
mostly located on the Atlantic sides of the South American imported sea lion, all preventative measures and equipment
and African continents, and the Pacific Ocean around should be used by the staff that performs the necropsy,
Australia and New Zealand.18 because this procedure carries a higher risk for exposure
and contamination of human participants, especially in
Epidemiology large mammals. For pinnipeds, typical M. pinnipedii gross
lesions consist of granulomas within lymph nodes (cervical,
The infection is likely to remain dormant for several years submandibular, tracheal, mediastinal, and mesenteric)19,24
in pinnipeds, with no clinical signs detected. Among marine and/or organs such as lung, spleen, liver, uterus,25 and
animals, the South American sea lion (Otaria byronia) seems bladder. In pregnant animals, lesions may be present in the
to have the greatest risk of spreading infection, mainly placenta.24 Granulomas may have a soft or thicker core and
because of sequential importation of infected wild animals calcifications may be present in the center and felt while
in the 1980s and early 1990s from Chile and Uruguay cutting tissue. Every suspected lymph node (either affected
to European zoos. The different outbreaks noticed in or draining a suspected area) and organ should be sent for
captivity since 1998 are often retrospectively found to cytology (either Ziehl-Neelsen or Auramine stains), PCR,
share wild-born imported pinnipeds as an index case or and culture.
as part of the affected group. Transmission mainly occurs Gross lesions of M. pinnipedii infections could not be
via direct contact (within rookeries in the wild, common differentiated from those of other MTBC species (e.g., M.
resting places in zoos or aquariums), water (oral route), bovis)26 or those of nontuberculous mycobacteria (NTM)
and aerosols. Transmission to other species followed the that can also affect pinnipeds (e.g., M. avium).26,27
From Bastida R, Guichon R, Quse V: Escenarios para el origen y dispersión de la tuberculosis en Patagonia Austral y Tierra del Fuego. Nuevos actores y líneas de
evidencia. In: XVII Congreso Nacional de Arqueología, 11-15 de octubre de 2010, Argentina, Mendoza, pp 227–230.
CHAPTER 85 Mycobacterium pinnipedii 605
Spoligotype pattern
Isolate/strain/type 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43
M. tuberculosis H37Rv
M. africanum TMC 12
M. bovis BCG
M. bovis AN5
M. microti lama type
M. microti vole type
Number of isolates W:Wild C:Captive
Australia (W+C) Argentina Uruguay (W) United Kingdom (C) New Zealand (W) Total
M. pinnipedii SS-1 (SB0155) 15 (A. sp) 13 (O. b) 28
M. pinnipedii SS-2 (SB2480) 1 (O. b) 3 (G. g, L. l, A. sp, T. sp) 1
M. pinnipedii SS-3 (SB0659) 3
M. pinnipedii SS-4 (SB0162) 3 (A. sp) 3
European Zoos outbreaks (all captive) with years of occurrence
ZOO Germany 1 Zoo France 2 Zoo France 1 Zoo Netherland 1 Zoo France 3 Zoo France 4
M. pinnipedii SS-1 (SB0155) 2006 (O. b) 1992–1993 (P. sp, O. b) 2006 (O. b) 2007 (O. b)
M. pinnipedii SS-5 (SB1162) 2001–2006 (C. b, T. sp, O. b) 2004–2006 (T. sp)
M. pinnipedii SS-6 (SB2479) 2009 (O. b)
• Figure 85.1 Spoligotype patterns of several Mycobacterium tuberculosis complex species and those
of M. pinnipedii strains gathered so far from the field (both in captive and natural settings). Black and
blue squares represent the presence or the absence of an individual spacer sequence, respectively. Years
Species affected in brackets for each outbreaks: A. sp, Arctocephalus spp.; G. g, Gorilla; L. l: Lama;
O. b, Otaria byronia; P. sp, Panthera spp.; T. sp, Tapirus spp. Most represented spoligotype is the SS-1
(SB0155).
606 SE C T I O N 17 Marine Mammals
of a selected biomarker, either cellular or serologic, is always antigen expression9 in M. pinnipedii has already been used
unpredictable and none of the current studies or reports to differentiate it from all other MTBC species that produce
were able to detect a theoretical profile of immune reaction this antigen (especially M. bovis). ELISA,38 multiantigen
sequence, as could be done for cattle. print immunoassays (MAPIAs)24,31 and lateral flow system
hand kits (STAT PAK and Dual Path Platform [DPP],
Cellular Immunity Exploration Chembio Diagnostic Systems, Inc., Medford, NY) have
Cellular immunity may be assessed through intradermal already been tested with various levels of success, but without
testing; intradermal injection of tuberculin induces a consistent validated sensitivity and specificity. Regarding
type IV hypersensitivity within 24–72 hours, which can serologic assays, feedback from the field seems to show
be measured by a local reaction. In cattle, M. pinnipedii that DPP testing is more sensitive than STAT PAK,25 for
infection was proven to induce a positive intradermal test example, 58% for STAT PAK versus 87.5% for DPP on a
performed with a purified protein derivative (PPD) dose sample of 10 cases.24 However, none of these tests have been
of 5000 UI/0.1 mL.22 In other terrestrial mammals, use of studied with a large enough sample comparison to indicate
a comparative intradermal test with M. avium PPD as a a gold standard, and therefore serologic results should be
comparison reagent was sometimes also applied to detect interpreted with caution. At the date of writing this chapter,
M. pinnipedii infection in tapirs,28,34 but with an apparent only DPP remains available to the veterinary practitioner;
lack of sensitivity and low negative predictive value. In MAPIA and STAT PAK have been discontinued.
pinnipeds, the skin test was also used with medium to poor The aquatic environment of pinnipeds is likely to expose
success.4,6,24,35 Although the recommended skin test sites them to many waterborne environmental mycobacteria and
may be the neck or flipper skin, the specific physiology of this may interfere with any indirect examination outcome,
pinnipeds may explain this failure because skin test efficiency because gene coding for ESAT-6-like proteins and CFP-10-
mostly relies on the population of T lymphocytes first, like proteins were demonstrated in several NTMs such as
and then macrophages within the first skin layers: aquatic M. kansasii, M. marinum, M. szulgai, M. flavescens, M.
mammals such as pinnipeds have developed vasoconstriction gastri, and M. smegmatis.39 However, the expression rate of
and dilation abilities for diving and temperature regulation these genes in those NTMs is questionable and homology of
that may totally impair the hypersensitivity sequence, from these proteins differs. Therefore, specificity of the humoral
none (apparent anergy) to overreaction (Koch reaction), test will be based on the test technology and qualitative/
which may induce flipper necrosis. quantitative approach to those MTBC proteins. Hence,
The use of interferon gamma release assays (IGRAs) cross-reaction should not be an immediate argument for
could be considered in validated species like cattle because antibody titers, whereas a single seropositivity with one
the MTBC antigens used in IGRAs (bovine PPD, ESAT-6, serologic assay must not be associated with certain infection
CFP10) are also shared by M. pinnipedii and could work as but should raise concerns and initiate deeper screening.
immunostimulants for lymphocytes. However, apart from
cattle, this is not a currently available option, for example, Interferences Between Cellular and Humoral Tests
for pinniped screening, mainly because there are no kits When designing a diagnostic and examination plan, the
available with a pinniped-specific interferon enzyme-linked veterinarian should know that application of an intradermal
immunosorbent assay (ELISA). However, the concept of test may have an impact on subsequent humoral tests. Injec-
detecting mRNA sequence coding for interferon, instead tion of antigens such as PPD could provoke a nonspecific
of interferon itself,36 could theoretically be applied to pin- immune reaction to several hundred antigens contained
nipeds, because these sequences are very similar among in the PPD and artificially induce antibody titers that are
carnivores. unrelated to the real infection by M. pinnipedii (A. Lécu
and G. Lacave, personal communication, 2012), but may
Humoral Immunity Exploration remain for months.
Pinnipeds and terrestrial mammals may produce detectable Injection of tuberculin has already been used as a “booster
antibodies during the course of M. pinnipedii infection.24,34 effect” in other animals30,36 (including humans) to trigger
Although theoretical mycobacteria immune response pre- an anamnestic rise of antibody in latent infected individuals
dicts low antibody titers at the beginning of an infection with low preexisting immune response. However, consider-
course37 and a rise in humoral response when mycobac- ing the nonspecific humoral reaction that may happen with
teria are replicating, spreading, and invading organs (i.e., PPD injections and considering that the booster effect was
antigens become uncovered), serologic profiles may follow not assessed in most zoo species that are already listed as
an unpredictable timeline pattern at the individual level. hosts for M. pinnipedii, this procedure is not recommended
M. pinnipedii expresses the same immunostimulant anti- because it is likely to increase false positive occurrence.37
gens as other members of the MTBC, especially ESAT6,
CFP10, and MPB83 (although the latter is not expressed Treatment
in M. pinnipedii)15: there is currently no known antigen/
antibody that is specific to M. pinnipedii. However, while Depending on a country’s animal health laws (M. pinnipedii
the sequence does exist in its DNA, the absence of MPB70 is not considered in European Union Animal Health Law,
CHAPTER 85 Mycobacterium pinnipedii 607
which focuses on M. bovis) and risk analysis, treatment biofilms that coat pipes or filtration media (sand). Hence,
could be an option for suspicion, contact (i.e., prevention), a sterilization phase is required and the chosen disinfection
or even confirmed positive animals. usually includes one or a combination of the following
M. pinnipedii is usually sensitive to typical antituberculous items: ultraviolet (UV) light, chlorine, and ozone.
drugs, at least in vitro,9,40 but antibiograms for strains found To decrease the MTBC load by 3–5 logs, UV devices
circulating in zoos (see Fig. 85.1) were rarely performed. should be set around 254 nm wavelength and should
Reported treatment in pinnipeds relied on bitherapy24 maintain power over 10–15 mJ/cm2. Although these are
or tritherapy41 with use of oral rifampicin (7.5 mg/kg), the usual settings applied in zoo and aquarium LSSs,
isoniazid (5 mg/kg), and addition of ethambutol (15 mg/ the age of the lamp, the turbidity of the water, and the
kg). There are no reported pharmacokinetic studies of any actual percentage of whole filtered volume passing through
antituberculous drugs in pinnipeds, and hence these dosages UV devices are all factors that significantly decrease UV
do not necessarily result in therapeutic plasma levels. More- efficiency against MTBC species, thus leading to longer
over, several major side effects were noted, such as anorexia duration of mycobacteria in the water.
(occurring 1–2 weeks after treatment began), abdominal Chlorine is not considered effective in removing
discomfort, lethargy, and suspected hepatotoxicity.41 mycobacterium from water in which animals reside: in the
These side effects are likely to impair compliance and concentrations of free chlorine that are compatible with
then to increase all risks associated with suboptimal anti- pinniped health (below 0.7 ppm),38 there is no bactericidal
biotic levels, such as persistence of shedding and resistance effect against mycobacteria, which require at least 1.0 ppm
acquisition; persistence or relapse of infection have been for more than 8 hours to decrease load by 5 logs.43 For
noted in Patagonian sea lions 11 months after treatment both UV and chlorine, organic aggregation that commonly
initiation.24 Therefore, a decision to treat should be appro- occurred in marine mammal water actually impaired the
priately balanced against euthanasia based on an exhaustive disinfection level by harboring mycobacteria.44
risk analysis including access to animals, medical training Thus, ozone remains the most effective tool for eradicat-
levels, ability to follow plasma levels, and all transmission ing M. pinnipedii because the contact time value (a product
opportunities (staff and other mammal species). of the concentration and contact time) effective range for
killing 99.9% of mycobacteria is within the range that is
customarily applied in aquarium and marine mammal LSS
Prevention contact chambers (0.06 mg/min/L).45
Animals The destination of water waste from an LSS (filter back-
wash) should also be monitored because it may move M.
Considering all antemortem diagnostic difficulties men- pinnipedii farther in or out of zoo or aquarium enclosures.
tioned previously, the examination panel to be included
in quarantine of any incoming pinniped should first relay Husbandry: Good Staff Practices
the animal’s history, especially a history of any contact with
high-risk animals such as imported wild contact specimens. By default, pinnipeds should be considered at risk for
According to this history, serologic assays and PCR on M. pinnipedii, and thus management practices should be
bronchoalveolar lavage can be recommended, along with adjusted to avoid hazardous actions or procedures that may
a CT scan and any other relevant clinical assessment in lead to transmission. Regarding staff and visitors, close-
case of higher risk. Repetition of examination is highly contact exercises, especially mixing aerosol production
recommended considering intermittent shedding and (“kiss,” “blow,” or even “bark to face”) should be avoided
immunologic status variations. as much as possible. Regarding animal management,
high Pinnipedii-pressure hose use should be restricted to
Husbandry: Water a minimum and the direction of the produced aerosol
should be monitored when used. Any devices in contact
Little is known about M. pinnipedii survival rate in the with pinnipeds or water should be regularly cleaned and
environment,42 but as a member of the MTBC, it can exposed to sunlight; especially, all tank cleaning and diving
likely survive outside the host organism for a long time; equipment should be disinfected because they are perfect
this “compensates” for its limited ability to multiply outside support venues for biofilm persistence. Keepers and trainers
the host organism. Even though they do not sporulate, should not care for other mammal species during their day,
MTBC species can usually still be cultured from a damp but if this is necessary, strict hygienic measures must be
environment protected from direct sunlight after the lapse applied (foot bath, different clothes, and tools).
of several months or years.42 Finally, health monitoring of marine mammal keepers
A life-support system (LSS) can be thought of as a primary and trainers in zoos, aquariums, or rehabilitation centers
defense against mycobacterium persistence and transmis- should be promoted,46 and attention must be focused on
sion within water. Mechanical and biological filtration are their preexisting status for tuberculosis. In case of a suspi-
not efficient in removing MTBC organisms and may even cious or confirmed case, the physician can recommend a
create a hidden burden of mycobacteria, e.g., embedded in skin test and/or IGRAs21 to screen for exposure; knowledge
608 SE C T I O N 17 Marine Mammals
of initial staff status regarding these tests is important, 16. de Amorim DB, Casagrande RA, Alievi MM, et al: Mycobacte-
especially in deciding whether to administer prophylactic rium pinnipedii in a stranded South American sea lion (Otaria
antituberculous treatment. byronia) in Brazil, J Zoo Wildl Med 50:419–422, 2015.
17. Bigi F, Garcia-Pelayo MC, Nunez-Garcia J, et al: Identification of
genetic markers for Mycobacterium pinnipedii through genome
Acknowledgments analysis, FEMS Microbiol Lett 248(2):147–152, 2005.
18. Bastida R, Guichon R, Quse V: Escenarios para el origen y
The author would like to deeply thank all European dispersión de la tuberculosis en Patagonia Austral y Tierra del
Association of Zoo and Wildlife Veterinarians Tuberculosis Fuego. Nuevos actores y líneas de evidencia. In XVII Congreso
Working Group members, especially Maria-Laura Boschi- Nacional de Arqueología, 11-15 de octubre de 2010, Argentina,
roli from ANSES Maisons-Alfort, France. Mendoza, 2010, pp 227–230.
19. Jurczynski K, Lyashchenko KP, Gomis D, et al: Pinniped
References tuberculosis caused by Mycobacterium pinnipedii: Transmission
from aquatic to terrestrial mammals and diagnostic options. In
1. Rodriguez-Campos S, Smith NH, Boniotti MB, et al: Over- Proceedings of the AAZV, AAWV, AZA/NAG Joint Conference,
view and phylogeny of Mycobacterium tuberculosis complex Knoxville, TN, 2007, pp 74–75.
organisms: Implications for diagnostics and legislation of bovine 20. Moser I, Prodinger WM, Hotzel H, et al: Mycobacterium pin-
tuberculosis, Bovine Tuberculosis 97(Suppl):S5–S19, 2014. nipedii: Transmission from South American sea lion (Otaria
2. Bos KI, Harkins KM, Herbig A, et al: Pre-Columbian myco- byronia) to Bactrian camel (Camelus bactrianus bactrianus) and
bacterial genomes reveal seals as a source of New World human Malayan tapirs (Tapirus indicus), Vet Microbiol 127(3–4):399–406,
tuberculosis, Nature 514(7523):494–497, 2014. 2008.
3. Brites D, Gagneux S: Co-evolution of Mycobacterium tubercu- 21. Kiers A, Klarenbeek A, Mendelts B, et al: Transmission of (Myco-
losis and Homo sapiens, Immunol Rev 264(1):6–24, 2015. bacterium pinnipedii) to humans in a zoo with marine mammals,
4. Forshaw D, Phelps GR: Tuberculosis in a captive colony of Int J Tuberc Lung Dis 12(12):1469–1473, 2008.
pinnipeds, J Wildl Dis 27(2):288–295, 1991. 22. Loeffler SH, de Lisle GW, Neill MA, et al: The seal tuberculosis
5. Woods R, Cousins DV, Kirkwood R, et al: Tuberculosis in a agent, (Mycobacterium pinnipedii), infects domestic cattle in New
wild Australian fur seal (Arctocephalus pusillus doriferus) from Zealand: epidemiologic factors and DNA strain typing, J Wildl
Tasmania, J Wildl Dis 31(1):83–86, 1995. Dis 50(2):180–187, 2014.
6. Bernardelli A, Bastida R, Loureiro J, et al: Tuberculosis in sea 23. Nugent G, Cousins DV: Zoonotic tuberculosis in Australia and
lions and fur seals from the south-western Atlantic coast. Rev Sci New Zealand. In Zoonotic Tuberculosis, 2014, John Wiley &
Tech 15:985–1005, 1996. Sons, Inc, pp 221–234.
7. Thompson PJ, Cousins DV, Gow BL, et al: Seals, seal trainers 24. Jurczynski K, Lyashchenko KP, Scharpegge J, et al: Use of
and mycobacterial infection, Am Rev Resp Dis 147:164–167, multiple diagnostic tests to detect (Mycobacterium pinnipedii)
1993. infections in a large group of South American sea lions (Otaria
8. Bastida R, Loureiro J, Quse V, et al: Tuberculosis in a wild flavescens), Aquatic Mammals 38(1):43–55, 2012.
subantarctic fur seal from Argentina, J Wildl Dis 35:796–798, 25. Lacave G, Maillot A, Alerte V, et al: Atypical case of Mycobac-
1999. terium pinnipedii in a Patagonian sea lion (Otaria flavescens)
9. Cousins DV, Bastida R, Cataldi A, et al: Tuberculosis in seals and tuberculosis cases history review in captive pinnipeds. In
caused by a novel member of the Mycobacterium tuberculosis Proceedings of the International Association for Aquatic Animal
complex: Mycobacterium pinnipedii sp. nov, Int J Syst Evol Medicine, San Antonio, TX, 2009, IAAAM, p 157.
Microbiol 53(Pt 5):1305–1314, 2003. 26. Barnett JEF, Booth P, Brewer JI, et al: Mycobacterium bovis infec-
10. Ahmed N, Alam M, Abdul Majeed A, et al: Genome sequence tion in a grey seal pup (Halichoerus grypus), Vet Rec 173(7):168,
based, comparative analysis of the fluorescent amplified fragment 2013.
length polymorphisms (FAFLP) of tubercle bacilli from seals 27. Foster G, Stevenson K, Reid RJ, et al: Infection due to Myco-
provides molecular evidence for a new species within the Myco- bacterium avium subsp. avium in a free-ranging common seal
bacterium tuberculosis complex, Infect Genet Evol 2(3):193–199, (Phoca vitulina) in Scotland, J Wildl Dis 49(3):732–734, 2013.
2003. 28. Gomis D, Moisson P, Lyashchenko KP, et al: Tuberculosis
11. Tortoli E: The new mycobacteria: an update, FEMS Immunol outbreaks in Mulhouse Zoo, 1992–96 and 2005–06: diagnostic
Med Microbiol 48(2):159–178, 2006. perspectives. In Proceedings of the 7th Conference of the Euro-
12. Kjeldsen MK, Bek D, Rasmussen EM, et al: Line probe assay pean Association of Zoo and Wildlife Veterinarians, Leipzig,
for differentiation within Mycobacterium tuberculosis complex. Germany, 2008.
Evaluation on clinical specimens and isolates including (Myco- 29. Jurczynski K, Scharpegge J, Ley-Zaporozhan J, et al: Computed
bacterium pinnipedii), Scand J Infect Dis 41(9):635–641, 2009. tomographic examination of South American sea lions (Otaria
13. Forrellad MA, Klepp LI, Gioffré A, et al: Virulence factors of flavescens) with suspected Mycobacterium pinnipedii infection,
the Mycobacterium tuberculosis complex, Virulence 4(1):3–66, Vet Rec 169(23):608, 2011.
2013. 30. Lécu A, Ball R: Mycobacterial infections in zoo animals: relevance,
14. Mostowy S, Inwald J, Gordon S, et al: Revisiting the evolu- diagnosis and management, Int Zoo Yearb 45(1):183–202, 2011.
tion of Mycobacterium bovis, J Bacteriol 187(18):6386–6395, 31. Arbiza J, Blanc A, Castro-Ramos M, et al: Uruguayan pinnipeds
2005. (Arctocephalus australis and Otaria flavescens): evidence of influ-
15. Alito A, Romano MI, Bigi F, et al: Antigenic characterization enza virus and Mycobacterium pinnipedii infections. In Romero
of mycobacteria from South American wild seals, Vet Microbiol A, Keith EO, editors: New approaches to the study of marine
68(3–4):293–299, 1999. mammals, 2012, Publisher: InTech, pp 151–183, (Chapter 7).
CHAPTER 85 Mycobacterium pinnipedii 609
32. Caulfield AJ, Wengenack NL: Diagnosis of active tuberculosis 40. Bernardelli A, Morcillo N, Loureiro J, et al: In vitro suscep-
disease: from microscopy to molecular techniques, JouJ Clin tibility testing of Mycobacterium tuberculosis complex strains
Tuberc Other Mycobact Dis 4:33–43, 2016. isolated from seals to antituberculosis drugs, Biomedica 24(Suppl
33. Tryland M, Nesbakken T, Robertson L, et al: Human pathogens 1):85–91, 2004.
in marine mammal meat - a northern perspective, Zoonoses Public 41. Guglielmi E, Biancani B: Two suspected cases of tuberculosis
Health 61(6):377–394, 2014. in captive Patagonian sea lions (Otaria flavencens) in Italy. In
34. Jurczynski K, Lyashchenko KP, Gomis D, et al: Pinniped tuber- Proceedings of the 44th Annual International Association for
culosis in Malayan tapirs (Tapirus indicus) and its transmission Aquatic Animal Medicine Conference, Marine Mammal Center
to other terrestrial mammals, J Zoo Wildl Med 42(2):222–227, Sausalito, CA, 2013.
2011. 42. Kazda J, Pavlik I: Obligate pathogenic mycobacteria. In Kazda
35. Kriz P, Kralik P, Slany M, et al: Mycobacterium pinnipedii in a J, Pavlik I, Falkinham JO, III, et al, editors: The ecology of myco-
captive Southern sea lion (Otaria flavescens): a case report, Vet bacteria: Impact on animal’s and human’s health, Berlin, Germany,
Med (Praha) 56:307–313, 2011. 2009, Springer, pp 13–20.
36. Lécu A, Ball R: Recent updates for antemortem tuberculosis 43. Pelletier PA, du Moulin GC, Stottmeier KD: Mycobacteria in
diagnostics in zoo animals. In Fowler’s zoo and wild animal public water supplies: comparative resistance to chlorine, Micro-
medicine (vol 8). 2015, pp 703–710. biol Sci 5(5):147–148, 1988.
37. Lynch M, Nielsen O, Duignan PJ, et al: Serologic survey for 44. Bohrerova Z, Linden KG: Ultraviolet and chlorine disinfection
potential pathogens and assessment of disease risk in Australian of mycobacterium in wastewater: effect of aggregation, Water
fur seals, J Wildl Dis 47(3):555–565, 2011. Environ Res 78(6):565–571, 2006.
38. Barnes J, Higgins D, Gray R: Pinnipeds. In Vogelnest L, Woods 45. Langlais B, Reckhow DA, Brink DR: Practical application of
R, editors: Medicine of australian mammals, Collingwood, Vic- ozone. In Ozone in water treatment—application and engineering,
toria, Australia, 2008, CSIRO Publishing, pp 541–589. Denver, CO, 1991, American Water Works Association Research
39. van Ingen J, de Zwaan R, Dekhuijzen R, et al: Region of difference Foundation, pp 221–222.
1 in nontuberculous Mycobacterium species adds a phylogenetic 46. Australian Wildlife Network Seal Tuberculosis in Australia. Fact
and taxonomical character, J Bacteriol 191:5865–5867, 2009. Sheet. [www.wildlifehealth.org.au], 2010.
86
Lens Diseases and Anesthetic
Considerations for Ophthalmologic
Procedures in Pinnipeds
CARMEN M.H. COLITZ AND JAMES E. BAILEY
610
CHAPTER 86 Lens Diseases and Anesthetic Considerations for Ophthalmologic Procedures in Pinnipeds 611
Exposure to ultraviolet (UV) light in the form of sun- completely resolve, the chronicity of this disease likely
light or other forms of radiation is a well-documented cause causes the continuous subclinical uveitis contributing to
of cataracts in many species.8–11 In a cross-sectional study, cataract formation, lens instability, and eventual luxation.
100% of all species who had excessive exposure to sunlight It has been suspected that similar risk factors to those of
had cataracts, and pinnipeds that did not have any access lens diseases might be to blame for keratopathy. A cross-
to shade were almost 10 times more likely to develop lens sectional study of 319 pinnipeds around the world found
diseases.1 that UV Index less than 6, not having had eye diseases
The risk factor regarding history of fighting would indi- or trauma, being older than 10 years of age, and having
cate a possible traumatic incident contributing to cataract been tested for Leptospirosis were risk factors for having
formation. Therefore managing more aggressive animals pinniped keratopathy.14 Most of these factors were similar
properly is indicated. The risk factor regarding previous to those affecting cataract incidence.
eye disease, most likely keratopathy, will be discussed later. UV Index was not evaluated in the initial cataract study,
Because younger animals were also affected, there may although it would likely be significant because it was in
be a genetic predisposition to cataracts in some lines of the cornea study. The UV Index was established by the
pinnipeds. Cataracts have been diagnosed in numerous World Health Organization for human sun exposure, and
generations of related animals in some facilities (Colitz, per- the United States has created a scale that conforms to these
sonal observation). With better data collection and genetic guidelines.15 Even a UV Index of 1 or 2, although “low,”
testing, there may be a means to evaluate this hypothesis can cause sunburns in individuals who easily burn; therefore
in the future. using broad-spectrum sunscreen and sunglasses are sug-
gested. A UV Index of 3–5 is “moderate” in risk of harm
Concurrent Eye Problems from the sun. Even levels less than the number found to be
risky to pinnipeds in this study (i.e., 6) can potentially cause
The most common concurrent eye problem in pinnipeds harm to eyes if sun protection is not worn. The UV Index
is keratopathy. Keratopathy had initially been described of 6–7 is in the high-risk range, and there are areas of the
in otariids12; however, it occurs in all species evaluated world with UV Indices higher than this. These animals, if
to date and is better termed pinniped keratopathy (Fig. without proper shade structure or other UV Index–lowering
86.1). Interestingly, it does not appear to occur in pin- means, had chronic unrelenting keratopathy. Pinnipeds and
nipeds in the wild, although it is possible and may not yet all marine animals that do not have shade structures or
be identified. means to lower the UV Index are at the highest risk for
Corneal diseases, including keratopathy, ulceration, both keratopathy and lens diseases.
and abscess formation, may cause secondary inflamma- Providing shade structures may be complicated because
tion or uveitis.13 Because keratopathy does not appear to the sun’s movements throughout the day may cause reflec-
tion rather than protection, depending on time of day and
season of the year. Other methods to lower the UV Index
include using UV paint colors that absorb UV light and/or
natural-appearing pool walls and bottoms. Some facilities
have also allowed algae to grow on these surfaces as long as
it is controlled and devoid of dangerous microorganisms.
Surgical Considerations
Regardless of the cause, visually impairing cataracts and
painful anteriorly luxated lenses can be surgically removed.
This allows the animal to regain his or her previous quality
of life and relieves pain in those affected by uveitis and/
or anterior lens luxation(s). Surgical considerations include
overall health and wellness, although even elderly pinnipeds
have undergone successful surgical lensectomy.
Surgical lensectomy for cataracts and/or lens luxation
has become one of the most commonly performed surgical
procedures in otariids and phocids under human care or
at stranding facilities. There are two approaches for lens
removal, extracapsular and intracapsular. Extracapsular
cataract surgery is when the anterior lens capsule is opened
• Figure 86.1 The left eye of a California sea lion (Zalophus california-
nus) with stage 3 pinniped keratopathy. There is diffuse corneal edema, (capsulotomy) and the lens cortex and nucleus are removed;
stromal loss, and temporal limbal hyperemia. Blepharospasm was this can be performed either via phacoemulsification or via
alleviated using topical anesthesia for examination and photography. manual extraction of the lens and nucleus. In the author’s
612 SE C T I O N 17 Marine Mammals
experience, all pinnipeds with cataracts that were older than medical behaviors may also reduce the need for mechanical
2 years so far have had lenses so dense or unstable that restraint devices such as netting, slings, and squeeze cages.
phacoemulsification has not been practical or possible. After Unfortunately, excessive reliance on mechanical restraint
the lens cortex and nucleus are removed via a 160-degree may lead to physical injury to the patient or eye, hyper-
limbal corneal incision, the remaining cortical material is thermia, or exertional myopathic conditions.18 With proper
flushed out of the capsule and posterior chamber and out desensitization training and chemical sedation, mechanical
of the eye; then the lens capsule is gently manipulated and devices may be used to improve the safety of anesthetic
most are removed completely. Yearling phocids and otariids procedures for both the patient and the animal handlers,
have undergone phacoemulsification because their lenses without undue added risk.
are softer.16 There is a report of a fur seal that underwent Although anesthesia of pinnipeds has been induced with
phacofragmentation,17 and the author explained that the inhalant anesthetic by mask under behavioral control,19 a
lens was so dense that he had to break it into quadrants balanced anesthetic approach is more appropriate. Balanced
and then manually remove the pieces via a larger incision; anesthesia involves the use of a combination of drugs, with
the lens material was too dense for traditional small incision each drug delivered in an amount sufficient to produce
phacoemulsification (J. S. Smith, personal communica- the desired effect while keeping undesirable effects to a
tion). Many cataractous lenses are clinically luxated into minimum. Inhalation anesthetics delivered alone will
the anterior chamber; these are removed via intracapsular produce excessive cardiopulmonary depression and no anal-
lens extraction wherein the lens capsule is not opened gesia. This does not imply that the practitioner cannot induce
and the entire lens (i.e., lens capsule, cortex, and nucleus) anesthesia by mask under behavioral control, but it would
is removed. be wise to supplement the anesthetic event with analgesic
agents, muscle relaxants, or other appropriate inhalation
anesthetic–sparing drugs after the pinniped is intubated
Anesthesia of Pinniped for and monitored. The benzodiazepines, midazolam and diaz-
Ophthalmologic Procedures— epam, and/or the opioids, butorphanol and meperidine,
an Overview have commonly been used for this purpose in pinnipeds.
Alpha-2 agonists have also been used in pinnipeds but have
Here we cover concepts as they relate to anesthesia of proven problematic for geriatric ophthalmologic patients.
pinnipeds for ophthalmologic procedures. The first step Alpha-2 adrenergic agonists, such as medetomidine and
in the process is determining whether immobilization dexmedetomidine, will provide profound sedation, analge-
(general anesthesia) is necessary to perform the procedure. sia, and muscle relaxation and are reversible. Unfortunately,
In general, anesthesia will be necessary for ophthalmologic they also cause bradycardia, interfere with contractility of
surgical procedures of pinnipeds. the heart, and cause peripheral vasoconstriction leading to
A voluntary preanesthetic health assessment provides increased afterload. Unsurprisingly, alpha-2 agonists also
a general overview of the animal’s current health status reduce renal blood flow in healthy subjects under optimal
prior to anesthesia in managed populations where medical conditions, even at low dosages. In addition, the alpha-2A
behaviors can be cultivated. Where possible, this should adrenoceptor is the alpha-2 adrenoceptor subtype primarily
include complete blood count, serum chemistry panel, and involved in the regulation of blood glucose homeostasis.
physical examination. Further information can be garnered Alpha-2 agonists should be considered a poor choice in
from urinalysis, radiographs, ultrasound, and echocardiog- the patient with poor glycemic control. The reversibility
raphy, where appropriate. Unfortunately, changes in vision, of alpha-2 agonists is enticing; however, sudden reversal
as well as pain associated with many eye problems, may lead and reduction in afterload could lead to cardiovascular col-
to loss of useful medical behaviors. Preoperative analgesia lapse in patients with limited cardiovascular reserve, such
may be necessary, and trainers must be vigilant to maintain as the geriatric patient. Furthermore, a historically high
necessary medical behaviors. Given that ophthalmologic negative outcome rate in ophthalmologic cases receiving
surgical procedures are often performed on geriatric patients medetomidine or dexmedetomidine has been observed by
in managed populations, the preanesthetic health assess- the authors, with the development of excessive perioperative
ment is imperative to identify other potential underlying intraocular hemorrhage leading to hyphema in pinnipeds
disease states. in which alpha-2 adrenergic agonists (or the reversal agent
Anesthesia of pinnipeds requires a balance of behaviors, atipamezole) have been used (Colitz, personal observation).
mechanical restraint, and chemical restraint. The practitioner For these reasons, the alpha-2 agonists are considered a
must consider the patient species, size, age, demeanor, and poor choice in many geriatric pinnipeds undergoing oph-
training, as well as the working environment, to best tailor thalmologic procedures, despite their reversibility. However,
the anesthetic protocol. Darting an animal with ultrapotent the alpha-2 adrenergic agonists are a tool of consideration
anesthetic agents to cause immobilization (anesthesia) may for use in young, healthy pinniped subjects in which it is
result in severe negative physiologic effects. Desensitiza- necessary to rely more on chemical restraint for the safety
tion training and voluntary medical behaviors contribute of human personnel. The practitioner will need to carefully
in no small part to positive outcomes. These voluntary weigh the risks and benefits of this choice.
CHAPTER 86 Lens Diseases and Anesthetic Considerations for Ophthalmologic Procedures in Pinnipeds 613
Rapid sequence induction of anesthesia with intravenous towel-drying after induction is helpful in limiting surface
(IV) agents is performed in cases in which venous access has evaporative cooling. Warming of replacement fluids gener-
proven feasible under mild sedation. Propofol IV, with or ally has an absolute minimal effect on patient warming;
without additional midazolam IV, can be delivered in the however, delivery of cold fluids is of no benefit. Several
extradural vein of phocids and odobenids. Both vascular types of patient-warming devices are available. The most
access and behavioral control are often more complicated effective systems are likely the warm circulating air-blanket
in otariids, and otariids are less likely than phocids to systems. The need for warming will depend on the working
breath-hold during inhalation anesthetic delivery by mask, environment, but the majority of patients in an appropriate
so mask induction is more often used for otariids. Both of surgical environment will arrive normothermic and quickly
the former methods of induction may become unnecessary become hypothermic after induction of anesthesia if the
when patients are darted with ultrapotent drug combina- presurgical environment is temperature controlled to the
tions to “immobilize” (anesthetize) the patient, but such needs of the surgeon and the presurgical process does not
combinations are not recommended here. lead to exertion. It is important to remember that the dosage
Vascular access is feasible in pinnipeds. Venipuncture of of inhalant anesthetic gas necessary to maintain anesthesia
the extradural vein of phocids and odobenids can be con- is reduced by hypothermia and recovery is prolonged by
verted to a small-diameter cannula using available epidural hypothermia. It is also important to know that cardiac
catheter kits. Venous access of the jugular vein of otariids contractility is reduced by hypothermia. Most patients do
is facilitated by ultrasound. Cannulation of pectoral flipper not benefit from hypothermia, and it should be avoided.
cephalic and brachial veins does not require ultrasound, Neuromuscular blockage is often needed for ophthal-
but ultrasound is a useful tool to facilitate access of these mologic procedures of pinnipeds to properly centralize
veins, as well as the pelvic flipper medial saphenous veins, and immobilize the eye for surgery. Atracurium, which
particularly in phocids. Standard over-the-needle catheters undergoes simple Hofmann degradation (elimination), has
can be used to reduce cost, but modern microintroducer kits been successfully used by the authors for neuromuscular
using a modified Seldinger method20 simplify and expedite blockade in phocids, otariids, and odobenids. Even though
cannulation (4–5F × 10 cm Stiffen cannula, nitinol mandrel easily eliminated from pinnipeds, any residual muscle weak-
stainless steel tip wire, echogenic needle, Micro-Introducer ness caused by atracurium can be antagonized by delivery
Kit, Innovative Veterinary Medicine, Ponte Vedra, FL). of edrophonium. Delivery of edrophonium should be done
Monitoring is often noninvasive (electrocardiogram, slowly, while still monitoring to avoid or detect bradycardia
pulse oximetry, capnography, temperature, and inspired/ that has been observed rarely in pinnipeds. The need for
expired gases), but invasive blood pressure monitoring has edrophonium may be assessed in part by use of nerve stimu-
become a standard of care for otariids, with limited success lators but is sufficiently imprecise to fully judge any residual
in phocids.21 Doppler flow probes and ultrasound facilitate paralysis. New true reversal agents, like sugammadex in
cannulation of the median artery of the pectoral flipper combination with rocuronium, will likely replace the use
or saphenous artery of the pelvic flippers of pinnipeds. of atracurium in pinnipeds in the near future.
Hypotension cannot be detected if it is not measured. Inhalation anesthetics depress the response of mammals
The purpose of monitoring blood pressure is to obtain to carbon dioxide (CO2). This depression of the response to
an objective measure of systemic circulation. Hypotension CO2 varies with species but does appear to follow the trend:
may be defined as a mean arterial blood pressure less than humans ≅ canid < equid < otariid ≅ odobenid < phocid.
60 mm Hg. Where anesthesia is the sole cause of hypoten- The shift in the response to CO2 is such that moderately
sion, the appropriate response would involve reducing the anesthetized otariids will not spontaneously ventilate until
level of anesthesia where possible, balancing the anesthetic the arterial partial pressure of carbon dioxide (PaCO2) is
technique with less cardiodepressive drugs (e.g., opioids), between 60 mm Hg and 80 mm Hg. However, it appears
volume loading where appropriate, inotropes such as the PaCO2 must rise very high—to the point of becoming an
dobutamine (0.2–2 µg/kg/min, IV; lower than terrestrial anesthetic itself (PaCO2 > 90 mm Hg)—in phocids; phocids
mammals) or ephedrine (0.05–0.1 mg/kg IV) and possibly will be unlikely to spontaneously ventilate even when only
vasopressors such as phenylephrine (1–3 µg/kg/min, IV) moderately anesthetized. Permissive hypercapnia involves
or norepinephrine (0.1–0.5 µg/kg/min, IV). Noninvasive accepting hypercapnia and associated acidemia to avoid
methods of estimating blood pressure have yet to be the potentially negative effects of mechanical ventilation.
validated, but the use of oscillometric cuffs on the pectoral Application of permissive hypercapnia has been accepted by
flippers of phocids has been used to follow trends in blood some in anesthesia of otariids, and its potentially beneficial
pressure. cardiovascular effects have been described. Before using
During anesthesia, body heat is lost through radiation, permissive hypercapnia, the effects of this therapy must
conduction, convection, and evaporation. Conductive and be understood. High blood levels and alveolar concentra-
convective heat loss can be minimized by various methods tions of CO2 could contribute to hypoxemia. Hypoxemia
of insulation. Evaporative heat loss is more difficult to can be remedied by administering supplemental oxygen,
control, such as losses to the breathing of anesthetic circuit which is commonplace. Hypercapnia may also cause tissue
dry gases, but dry-docking the patient prior to surgery or acidosis, resulting in decreased intracellular pH and altered
614 SE C T I O N 17 Marine Mammals
transmembrane electrolyte transport, glucose use, and surgery of marine mammals. This reflex is a well-known,
structure-function relationships of amino acids. Neuro- albeit rare, cause of cardiac arrest during eye surgery
logic sequelae of hypercapnia include sympathetic nervous of mammals. This reflex is observed during traction on
stimulation with release of epinephrine and norepinephrine, extraocular muscles, is greatly exaggerated in the presence
significant increases in cerebral blood flow, and elevation of of hypoventilation, hypoxemia, and acidosis, but might
intracranial pressures. Hypercapnia also leads to increased be prevented by a retrobulbar local anesthetic block or
cardiac output, decreased peripheral vascular resistance, administration of parasympatholytic drugs. The laryngeal/
and increased arterial blood pressure. However, as pH falls, pharyngeal reflexes can lead to bradycardia as well. It is well
left ventricular contractility is decreased (although cardiac established that general anesthetics modify the laryngeal
output may still be maintained). In addition, decreasing reflex, but, at light planes of anesthesia, activation of laryn-
pH causes a shift in the oxyhemoglobin dissociation curve geal vagal reflexes can occur during manipulation of the
to the right, promoting release of oxygen from hemoglo- airway. Adequate anesthesia prior to attempting intubation,
bin. However, cardiac arrhythmias are reported in some topical local anesthetic sprays, and parasympatholytic drugs
species with PaCO2 levels greater than 80 mm Hg. Type prevent this reflex.
I hearts (canid, felid, human) tend to be more prone to Controlled mechanical ventilation (CMV) is the current
such arrhythmias than type II hearts (equid, bovid, phocid, mechanical ventilation method available on nearly all vet-
otariid, and odobenid). If permissive hypercapnia is used, erinary mechanical ventilators. This CMV mode of ventila-
the PaCO2 should be maintained less than 80 mm Hg, the tion has often been treated as merely a convenience, taking
pH greater than 7.2, and the PaO2 well above 60 mm Hg. the place of squeezing the anesthetic reservoir bag by hand
A blood gas analyzer is mandatory for prompt, accurate to deliver a breath. However, the inconsistency of venti-
decision making. Although one may rationalize a degree lation associated with hand ventilation over long periods
of benefit of elevated PaCO2, recent work has suggested makes CMV more of a necessity than a mere convenience
that high levels of CO2 are not beneficial to pinnipeds and in ophthalmologic surgeries. CMV begins the ventilator
indicate a need for enhanced monitoring and support.22 cycle at a baseline pressure and elevates airway pressure to
Given these facts—as well as the use of neuromuscular deliver an appropriate tidal volume. Disadvantages to CMV
blocking agents—either manual or mechanical ventilation include the application of relatively higher-peak inspira-
is mandatory for ophthalmologic procedures. tory pressure and resultant higher mean intrathoracic pres-
It is well known from pinniped dive studies that pin- sure and reduced venous return and cardiac output, as well
nipeds demonstrate a profound “dive reflex” when strapped as poor distribution of inspired gas flow, with subsequent
to a board and forced to dive.23 It is also known that the ventilation-perfusion mismatch. The apneustic plateau
same animals, when allowed to dive voluntarily, do not ventilation (APV) mode used in early dolphin ventilation
demonstrate the same degree of dive reflex bradycardia as was designed in an attempt to mimic conscious marine
when forced into a dive.24,25 As such, there appears to be a mammal breathing patterns but still allowed for devel-
conscious stress component of forced dives (not knowing opment of atelectasis. Airway pressure release ventilation
when the dive will end). In none of these instances did the (APRV) was developed to improve ventilation of patients
subjects have a dive reflex leading to their death. There is with low compliance, and apneustic anesthesia ventilation
no evidence to suggest an animal will die directly from the (AAV) was designed to best normalize respiratory mechan-
dive reflex, although it will die if not allowed to surface ics of mammals under anesthesia. Both APRV and AAV
and breath. In addition, there is no evidence that the dive also mechanically mimic the breathing patterns of conscious
reflex remains intact during chemically induced uncon- marine mammals. The objective of mechanical ventilation
sciousness of pinnipeds. On the contrary, there is evidence of marine mammals is not to merely mimic the normal con-
that anesthesia depresses or eliminates the dive reflex.26 scious animal breathing pattern but to provide appropriate
There is also clinical evidence that heart rates of marine ventilation to unconscious marine mammals that are not
mammals stabilize under general anesthesia when proper spontaneously ventilating. The potentially beneficial modes
physiologic support is provided. Some reflexes, such as the of APRV and AAV ventilation for anesthetized pinnipeds are
laryngeal reflex and other vagally mediated reflexes, may under development and will be available in the near future.27
well be present at light planes of general anesthesia—such Assigning someone to provide oversight of all activi-
as evidenced at induction and recovery—and vary with ties related to the ophthalmologic surgical procedure—an
the anesthetic drugs used. These reflexes, or inadequate “incident commander”—ensures personnel, policies, and
respiratory support of the anesthetized marine mammals, procedures are optimized to improve emergency response,
may well be confused with a “dive reflex.” Indeed, severe especially when media become involved in reporting the
hypoxemia will lead to myocardial hypoxemia and terminal event. Quality staff should be informed and readily available
bradycardia but is not mediated by a reflex. to facilitate any potential emergency response. Assigning
Although a proper dive reflex may not be triggered an individual to focus solely on the management of the
during general anesthesia, the anesthetist should be aware of anesthetized patient is critical to prevent anesthesia-related
another related trigeminocardiac reflex—the oculocardiac problems. Given the potential for anesthesia-related
reflex—that can still be triggered during anesthesia for eye problems such as high CO2 levels, apnea, hypoxemia,
CHAPTER 86 Lens Diseases and Anesthetic Considerations for Ophthalmologic Procedures in Pinnipeds 615
A B
C D
• Figure 86.2 (A and B) Right and left eyes of an Australian sea lion (Neophoca cinera) with bilateral
mature cataracts. (C and D) Right and left eyes of the same Australian sea lion following bilateral
lensectomies.
bradycardia, thermoregulatory problems, and prolonged mild opacities (Fig. 86.3). Because lens luxations are
recoveries, it is important to assign an individual to focus severely painful in the acute stages, treatment with oral
solely on the management of the anesthetic event who and topical nonsteroidal antiinflammatory medications is
has a thorough understanding of intubation, mechanical important, along with topical hypertonic saline solution,
ventilation, vascular access, and drug delivery methods for topical carbonic anhydrase inhibitors to control elevated
the species undergoing the procedure. Understanding the intraocular pressure, and oral tramadol for control of pain.
potential complications of anesthesia, appropriate vigilance With time, the blepharospasm, epiphora, and photophobia
monitoring and being prepared to manage complications improve and may appear resolved, although these signs may
improves the overall outcome of ophthalmologic anesthetic wax and wane. It is doubtful that the pinniped is pain free,
procedures of pinnipeds (see also Chapter 29). and its stoic nature should not be cause for medications to
be ceased.
Surgical Candidates: Poor, Good, Great In eyes with chronic anterior lens luxations, the cornea
will be permanently opaque, although vision is improved in
Aside from overall health and wellness, the status of the eyes the majority of patients’ eyes in this condition. In addition,
is also important for predicting outcome. The best outcomes the pain is vastly improved following surgical lens removal.
occur in eyes that have visually impairing cataracts that are
not anteriorly luxated and that have not caused excessive Outcomes and Sequelae
chronic uveitis (Fig. 86.2). Following recovery, these eyes
will typically only have a limbal opacity or scar and the rest The ideal outcome is a pinniped patient that has resolution
of the cornea will be clear. of pain and regains sight. The majority of eyes have resolu-
Eyes that have acute lens luxation can regain almost tion of pain; however, causes of continued blindness include
complete transparency or, in some cases, only have residual severe keratopathy or retinal detachment.
616 SE C T I O N 17 Marine Mammals
A B
C D
• Figure 86.3 (A and B) Right and left eyes of a California sea lion (Zalophus californianus) with bilateral
anteriorly luxated mature cataracts. The corneas have vascularization, edema, and fibrosis secondary
to the lens luxations. (C and D) Right and left eyes of the same California sea lion following bilateral
lensectomies. The corneal fibrosis persists, but the animal’s pain has resolved and useful vision is restored.
References
Retinal detachments are uncommon in pinnipeds fol-
lowing cataract surgery. Animals that appear at increased 1. Colitz CMH, Saville WJA, Renner MS, et al: Risk factors
risk of retinal detachment are those that were stranded associated with cataracts and lens luxations in captive pinnipeds
young who already had cataracts or developed them soon in the United States and the Bahamas, J Am Vet Med Assoc
after being rescued. The vitreous in these animals is very 237:429–436, 2010.
liquefied, compared with the firm, clear, well-formed nature 2. Kern TJ, Colitz CMH: Exotic animal ophthalmology. In Gelatt
of normal pinniped vitreous material. Vitreal degeneration KN, Gilger BC, Kern TJ, editors: Veterinary ophthalmology, ed
is a predisposing factor to retinal detachment.28 5, Ames, IA, 2013, John Wiley & Sons, pp 1750–1819.
3. Stoskopf MK, Zimmerman S, Hirst LW, et al: Ocular anterior
Preexisiting retinal detachments are not a reason to avoid
segment disease in northern fur seals, J Am Vet Med Assoc
surgical lensectomy in pinnipeds. The removal of the lens 187:1141–1144, 1985.
will alleviate pain if luxated, and it will remove the risk of 4. Gerber JA, Roletto J, Morgan LE, et al: Findings in pinnipeds
luxation in the future in those eyes with cataracts. Lastly, stranded along the central and northern California coast,
cosmesis for the public may also be important in some 1984-1990, J Wildl Dis 29:423–433, 1993.
collection animals. If cosmesis is not as important, then 5. Colitz CMH, Gulland FMD, Palmer L, et al: Retrospective
enucleation may be performed. report of ophthalmologic problems in wild pinnipeds stranded
CHAPTER 86 Lens Diseases and Anesthetic Considerations for Ophthalmologic Procedures in Pinnipeds 617
along the California coast between 2005 and 2009, J Zoo Wildl 18. Bailey JE, Flanagan C, Meegan J, et al: Cogent evidence of
Med 2017. Accepted with revisions. rhabdomyolysis in a California sea lion (Zalophus californianus)
6. Hirst LW, Stoskopf MK, Graham D, et al: Pathologic findings and a South African fur seal (Arctocephalus pusillus pusillus)
in the anterior segment of the pinniped eye, J Am Vet Med Assoc during anesthesia. Proceedinsg of the 43rd Annual International
183:1226–1231, 1983. Association for Aquatic Animal Medicine Conference, Atlanta,
7. Stoskopf MK, Hirst LW: Examination of the anterior segment GA, 2012.
of the pinniped eye, Annu Proc Am Associ Zool Vet 64, 1991. 19. Haulena M: Otariid seals. In West G, Heard D, Caulkett N,
8. Colitz CMH, Bomser JA, Kusewitt DF: The endogenous and editors: Zoo animal & wildlife immobilizaiton and anesthesia,
exogenous mechanisms for protection from ultraviolet irradia- Ames, IA, 2007, Blackwell Publishing, pp 469–478.
tion in the lens, Int Ophthalmol Clin 45:141–155, 2005. 20. Seldinger SI: Catheter replacement of the needle in percutaneous
9. Truscott RJW: Human cataract: the mechanisms responsible; arteriography; a new technique, Acta Radiol 39:368–376, 1953.
light and butterfly eyes, Int J Biochem Cell Biol 35:1500–1504, 21. Bailey J: Arterial blood pressure monitoring of select pinnipeds:
2003. multi-case presentation. Proceedings of the 44th Annual IAAAM
10. Young SR, Sands J: Sun and the eye: prevention and detection Conference, 2013.
of light-induced disease, Clin Dermatol 16:477–485, 1998. 22. Ponganis PJ, McDonald BI, Tift MS, et al: Effects of inhalational
11. Chandler HC, Reuter KS, Sinnott LT, et al: Prevention of anesthesia on blood gases and pH in California sea lions, Mar
UV-induced damage to the anterior segment using Class I Mamm Sci 2017. [Epub ahead of print].
UV-absorbing hydrogel contact lenses, Invest Ophthalmol Vis Sci 23. Irving L, Scholander PF, Grinnell SW: Significance of the heart
51:172–178, 2010. rate to the diving ability of seals, J Cell Comp Physiol 18:283–297,
12. Colitz CMH, Renner MS, Manire CA, et al: Characterization of 1941.
progressive keratitis in Otariids, Vet Ophthalmol 13:47–53, 2010. 24. Thompson D, Fedak MA: Cardiac responses of grey seals during
13. Hendrix DVH: Diseases and surgery of the canine anterior diving at sea, J Exp Biol 174:139–154, 1993.
uvea. In Gelatt KN, Gilger BC, Kern TJ, editors: Veterinary 25. Kooyman GL, Ponganis PJ: The physiological basis of diving to
ophthalmology, ed 5, Ames, IA, 2013, John Wiley & Sons, pp depth: birds and mammals, Annu Rev Physiol 60:19–32, 1998.
1146–1198. 26. Whayne MD, Thomas F, Smith MDNT, et al: The effects of halo-
14. Colitz CMH, Saville WJA, Atkin C, et al: Epidemiological thane anesthesia on reflex cardiovascular responses to simulated
survey identifying risk factors for corneal disease in pinnipeds. diving and the Valsalva Maneuver, Anesthesiology 34:262–269,
Annual Conference of the International Association of Aquatic 1971.
Animal Medicine, 2013. 27. Bailey JE: Anesthesia ventilator for Atlantic bottlenose dolphins
15. Organization WH, editor: Global solar UV index a practical guide, and California sea lions: Navy STTR FY2014.A, 2014.
Geneva, Switzerland, 2002, WHO Library. 28. Vainisi SJ, Wolfer JC, Hoffman AR: Surgery of the canine
16. Esson DW, Nollens HH, Schmitt TL, et al: Aphakic phacoemul- posterior segment. In Gelatt KN, Gilger BC, Kern TJ, editors:
sification and automated anterior vitrectomy, and postreturn Veterinary ophthalmology, ed 5, Ames, IA, 2013, John Wiley &
monitoring of a rehabilitated harbor seal, J Zoo Wildl Med Sons, pp 1393–1431.
46:647–651, 2015.
17. Barnes JA, Smith JS: Bilateral phacofragmentation in a New
Zealand fur seal (Arctocephalus forsteri), J Zoo Wildl Med
35:110–112, 2004.
SECTION 18
Ruminants
87 Giraffe Husbandry and Welfare, 619
618
87
Giraffe Husbandry and Welfare
LAURIE J. GAGE
619
620 SE C T I O N 18 Ruminants
techniques as well as improved nutrition have been used to have responded favorably to the use of complex feeders
mitigate these problems (see also Chapter 88).10 that require the use of the tongue to access grain or hay.13
The addition of slatted tops to hay feeders eliminated oral
Complications of Anesthesia stereotypic behavior for some animals.13 Providing means
for giraffe to engage in more naturalistic foraging behavior
Giraffe tend to have more anesthetic-related complications and play behavior will improve their welfare. Each item
and deaths than other Artiodactyla because of their unique introduced into the giraffe enclosure should be carefully
anatomy and physiology, with most problems occurring scrutinized to ensure it is safe, because a number of giraffe
during induction and recovery.8 The use of a well-designed have experienced morbidity and mortality due to unfortu-
restraint device for induction, or a chute where a halter may nate accidents involving entrapment or entanglement with
be placed on a sedated giraffe to help control the head before enrichment items. Dangling enrichment, however safe it
induction, has helped prevent or mitigate injuries.8 While may seem, may best be offered only when staff members
anesthetic techniques have improved, applying training- are available to respond in case of entrapment. Buckets
based methods to obtain diagnostics such as radiographs or barrels with holes should be avoided or carefully evalu-
of the feet and limbs,10 blood collection, and maintenance ated to ensure giraffe cannot entrap an ossicone or other
procedures such as hoof trimming decreases the number body part.
of anesthetic events required to manage the animals and
reduces the chances of complications of anesthesia. Trauma
Social Structure Giraffe are prone to accidents that involve entrapment,
entanglement, slips, and falls. Giraffe have died because
Giraffe have a social structure, and individuals, especially their ossicones became entangled in hotwire, fencing, and
females, may show a preference for certain other individuals enrichment devices. Openings in doors, fences, or furnish-
within their herd.11 For optimal welfare of the animals, ings within an enclosure that a giraffe may put its head or
these social preferences should be considered when selecting ossicone through are potential hazards. Giraffe have hung
housing, planning movement between facilities, or plan- themselves in the crooks of trees, on pulley ropes designed
ning how individual animals will be combined or housed to open stall doors, and in the triangular truss elements
within the same facility. A study in the United Kingdom of installed to support a shade structure (Fig. 87.1). Vertical
seven giraffe collections involving 40 individuals described
bonds that are evident between animals, documenting
that giraffe seek out preferred partners, and that partner
preference is maintained over time.11 The strength of these
bonds must be considered when managing these animals.
Overcrowding or separating compatible individual animals
from each another may result in stress-related behavior. The
movement of individual female giraffe between herds has a
greater chance of being disruptive to these individuals than
when moving the males, and this should be a consideration
when determining the placement of animals for breeding
purposes.11
Adult males may exhibit intolerance of other males
within the same enclosure; however, serious altercations
between males are rare. Adult bulls may be housed with one
another or with younger males in the absence of females but
should be offered ample space, especially in night quarters.4
Enrichment
Giraffe are inquisitive and benefit from a variety of enrich-
ment items offered to them that will increase the amount of
complexity and stimulation within their environment and
provide opportunities for species-specific behavior.12 Enrich-
ment opportunities have improved the physiologic and
psychologic well-being of captive animals. For giraffe, these
may include puzzle feeders or elevated enrichment items that
encourage the development of play and manipulation with • Figure 87.1 Giraffe (Giraffa sp.) shade structure with openings
their tongues. Giraffe that exhibit oral stereotypic behavior large enough for the head or ossicone; may cause a fatal entrapment.
CHAPTER 87 Giraffe Husbandry and Welfare 621
posts spaced greater than 2 inches apart have proven deadly animals. They will ensure the barn heaters are free of flam-
to giraffe, as they may slide their jaw or neck between the mable material, such as bird nests, before the heaters are
posts and become entrapped. Grass growing on the outside activated, and will know to check the exhibit when there
of a livestock wire fence may entice a giraffe, resulting is rainfall or freezing conditions to eliminate the chance a
in entrapment in the fence in its effort to gain access to giraffe could slip or fall. Giraffe deaths have occurred from
the grass. Removing or mowing grass near fencing and preventable barn fires and toxic oleander browse. Ensure
covering any opening, including small keeper observation new keepers are adequately trained and work together with
windows or ventilation doors in the wall of a giraffe barn experienced personnel to ensure optimal welfare.
with bars or mesh with spaces less than 2 inches, will help
prevent giraffe head or ossicone entrapment. Materials on Training
the outside of giraffe enclosures should be placed such that
giraffe cannot reach them. One giraffe became entrapped in Using training techniques to address husbandry issues, such
orange plastic safety construction netting on the outside of as the movement of animals within a facility, or for hoof
its exhibit and died. Icy, wet, or slippery footing, especially maintenance, and to perform routine medical procedures,
where there is a slant or grade to the exhibit, has proven has been successful at many institutions housing giraffe.
fatal to giraffe. A faulty gate inside a giraffe barn allowed Training the animals not only helps limit the number of
incompatible animals into the same stall, resulting in the immobilizations necessary to conduct diagnostics or to treat
death of another animal. Keepers and management staff issues, such as overgrown hooves, but it also provides a form
should routinely inspect all giraffe areas, including areas of enrichment for the animals (see also Chapter 88).
outside of the enclosure that a giraffe might reach, and
ensure there are no holes, cracks, or other spaces large Acknowledgments
enough for a giraffe to put its head or ossicone through.
All ropes and wires within reach of a giraffe, including Thanks to Dr. Liza Dadone, Laurie Bingaman Lackey, Dr.
the keeper area, should be carefully evaluated to ensure Nora Wineland, Dr. Nicolette Petervary, and Dr. Elizabeth
entrapment is not possible. All heating elements should be Pannill for their assistance with this chapter.
checked routinely to ensure they are working, are clean and
free of flammable materials nearby such as nests, and they
must be installed and maintained properly. References
1. Dadone LI: Causes of mortality in zoo giraffes and okapi. In Pro-
Exhibit Design ceedings of the Midyear Conference of the American Zoological
Association, 2017.
Giraffe enclosures require thoughtful design to create 2. Gage LJ: Medical and husbandry risk management in giraffes:
an environment where there is adequate space for social updates on improving giraffe welfare. In Proceedings of the
interactions and where giraffe may carry out all of their Annual Conference of American Association of Zoo Veterinar-
natural behaviors. To prevent injury, ensure areas are free ians, 2013, pp 140–141.
of obstructions and the footing is reasonably level and 3. Potter JS, Clauss M: Mortality of captive giraffe (Giraffa
dry. Sloping exhibits may be hazardous. During cold and camelopardis) associated with serous fat atrophy: a review of
wet conditions, exhibits should be inspected to ensure five cases at Auckland Zoo, J Zoo Wildlife Med 36(2):301–307,
2005.
the giraffe do not encounter areas with frozen or slippery
4. AZA Giraffe Species Survival Plan. Giraffe Care Manual. Associa-
footing. Flooring in barns should be constructed using a
tion of Zoos & Aquariums, Silver Spring, MD (in progress).
nonslip material to ensure traction. A giraffe restraint device 5. Valdes EV, Schlegel M: Advances in giraffe nutrition. In Fowler
is highly desirable, and those designed to allow the giraffe ME, Miller RE, editors: Zoo and wild animal medicine, ed 7, St.
to pass thorough the device daily are optimal.4 Louis, MO, 2012, Elsevier, pp 612–618.
6. Clauss M, Rose P, Hummel J, et al: Serous fat atrophy and
Keeper Experience other nutrition-related health problems in captive giraffe
(Giraffa camelopardalis). An evaluation of 83 necropsy reports.
Experienced keepers are key to ensuring the health and In Proceedings of the European Association of Zoo and Wildlife
welfare of the animals. Giraffe do know their keepers, and Veterinarians, Budapest, Hungary, 2005, pp 233-235.
trust may be built, which is very important when training 7. Enqvist KE, Chu JI, Williams CA, et al: Dental disease and
serous atrophy of fat syndrome in captive giraffes (Giraffa
techniques are applied. Knowledgeable keepers are aware
camelopardalis). In Proceedings of the American Association of
of individual animal idiosyncrasies and will notice subtle
Zoo Veterinarians, 2003, pp 262–263.
changes in their behavior that will aid in early recognition 8. Bertelsen M: Giraffe. In Fowler ME, Miller RE, editors: Fowler’s
of stressful or medical conditions. They will notice and Zoo and wild animal medicine (vol 8). St. Louis, MO, 2015,
eliminate problems before they could cause injury or death Elsevier, pp 602–610.
to the animals. They are able to evaluate the quality and 9. Junge RE, Bradley TA: Peracute mortality syndrome of giraffe.
condition of the diet and know what browse items are In Fowler ME, editor: Zoo and wild animal medicine: current
appropriate and which may be toxic or harmful to the therapy, ed 3, Philadelphia, PA, 2003, Saunders, pp 547–549.
622 SE C T I O N 18 Ruminants
10. Dadone LI, Schilz A, Friedman SG, et al: Training giraffe 12. Shepherdson DJ: Introduction: tracing the path of environmental
(Giraffa camelopardalis reticulata) for front foot radiographs and enrichment in zoos. In Shepherdson DE, Mullen DJ, Hutchens
hoof care, Zoo Biol 35:228–236, 2016. D, editors: Second nature: environmental enrichment for captive
11. Rose P: Investigations in the social behavior of captive giraffe: animals, Washington DC, 2012, Smithsonian Institution Press.
what do the animals tell us about their life in the zoo? In 13. Fernandez LT, Bashaw MJ, Sartor RL, et al: Tongue twisters:
Proceedings of the International Association of Giraffe Care feeding enrichment to reduce oral stereotypy in giraffe, Zoo Biol
Professionals, San Francisco, CA, 2012. 27:200–212, 2008, doi:10.1002/zoo.20180.
88
Lameness Diagnosis and
Management in Zoo Giraffe
LIZA DADONE
L
ameness is a common health problem of adult giraffe opportunity to correct early changes of hoof shape before
(Giraffa spp.) at many zoos. Up to 80% of giraffe significant secondary pathology develops, leaving the giraffe
immobilizations are done to address hoof overgrowth with permanent hoof capsule distortions, which are chal-
and limping,1 indicating that lameness is an important lenging to correct once advanced (Fig. 88.1).
health problem for this species. Giraffe anesthesia can have Arthropathies are frequently associated with giraffe lame-
a 10% mortality rate,1 and some giraffe have died during ness. A variety of etiologies have been reported, including
anesthesia for hoof work.2 Due to risks associated with mycoplasma-associated polyarthritis,19 osteoarthritis,5,19,20
anesthesia, some cases are not diagnosed or treated until osteochondrosis,21 and septic and ulcerative arthritis.16 In
relatively late in the disease process. When lameness is severe a survey of European zoos, joint problems were more com-
or does not improve with treatment, giraffe are sometimes monly reported in front legs than in hind legs.11 In a study
humanely euthanized.3 This chapter describes diagnostics of the front feet of all 22 giraffe at one zoo, all animals had
and treatments to help improve lameness management for radiographic evidence of osteoarthritis by age seven in the
zoo giraffe and highlights the need for early intervention distal interphalangeal joint (coffin joint).5 This seems to be a
and preventative care. relatively early onset for a species with a maximum wild life
span of 22 years for males and about 28 years for females,22
Etiology but both age of arthritis onset and normal life span warrant
further study.
Multiple abnormalities have been described in giraffe that Fractures have also been relatively commonly described
may be associated with lameness. These include arthritis, in the distal phalangeal bone (pedal fractures) of the front
bone cysts,3,4 bruising, congenital deformities, dermatitis, foot.5,6,10,23,24 Many of these cases are associated with severe
fractures,4–8 hoof overgrowth,9–11 laminitis,3,11–13 ligament pedal osteitis and hoof overgrowth, with fractures at the site
injuries,8,12 nutritional imbalances, osteitis,5,10 osteochon- of the deep digital flexor tendon insertion along the weight-
drosis,14 osteolysis,4 osteomyelitis,3 pigmented villon- bearing axis of the foot (Fig. 88.2).5,10 Limb fractures at
odular synovitis,15 pododermatitis (“foot rot”),3 sole foreign other sites are anecdotally reported and generally have a
bodies,10,14 tenosynovitis,16 and trauma. While lameness is poor prognosis, except in relatively young animals.
often related to pain, neurologic or mechanical dysfunction Laminitis is likely underdiagnosed in zoo giraffe and
may also be an underlying cause.17 When multiple lesions may be either acute or chronic. On radiographs, lami-
are present, identifying the most clinically significant cause nitis present with a downward rotation of the digit, but
of lameness can be challenging. upwards rotation of the distal phalangeal bone has also
Hoof overgrowth is common in giraffe and may be been described.10,13 There are multiple possible etiologies for
associated with abnormal conformation,13 hypothyroid- laminitis, but suboptimal diet for this specialized browser
ism,18 insufficient exercise, nutritional imbalances, inap- is likely a factor for some giraffe. In one survey, facilities
propriate substrate, or trauma, among other causes.17 Hoof that reported laminitis cases fed higher proportions of
overgrowth changes weight distribution in the foot, which easily digestible feeds in the diet (bread, pure grains, fruits,
can then lead to secondary pathologies such as arthritis or vegetables) than facilities that did not report laminitis in
pedal osteitis.5,10 While regular hoof trims are the standard their giraffe.11 Further study is needed to better understand
of care for domestic horses, routine giraffe hoof trims have how rumen acidosis in giraffe, potentially caused by diets
been relatively uncommon, except at zoos that train for high in starch or low in fiber, may be related to some cases of
voluntary hoof work.10 For this reason, many zoos miss the laminitis.13
623
624 SE C T I O N 18 Ruminants
A B
C D
• Figure 88.1 Photos comparing zoo and wild giraffe suggest that hoof distortions may begin in some
zoo giraffe at a young age. Photos of the front foot of a 4-year-old female reticulated giraffe (Giraffa
camelopardalis reticulata) in a zoo and, based on size approximation, front feet from a similarly aged
young adult wild Nubian giraffe in Uganda. (A) In this zoo giraffe, the interdigital gap indicates interdigital
overgrowth. (B) The wild giraffe has a thinner interdigital space, and the hooves are longer and smoother.
(C) The sole of the zoo giraffe has asymmetric claw width, interdigital overgrowth, and a relatively hard,
flat sole. (D) The wild giraffe has relatively symmetric toe width, a concave foot shape, a markedly more
pliable sole surface, and longer toe tips.
• Figure 88.2 Positioning for giraffe front foot radiographs using trained behaviors. (A) In a restricted
contact setup, the giraffe is cued to position its foot on a hoof block, as close as possible to the radiograph
plate. For a dorsomedial-palmarolateral oblique view (DMPLO), the radiograph generator is positioned
at a 30° angle to the plate. (B) In the DMPLO radiograph image, this positioning partially separates the
paired digits of the foot. Radiographic lesions may include upward rotation of both distal phalangeal
bones, osteoarthritis of the distal interphalangeal joint (*), pedal osteitis of the solar margin, and an
articular fracture of the medial phalangeal bone (Δ). In this case, the fractured piece is similar in size to
the sesamoid bones and is at the site of the deep digital flexor tendon attachment. (C) Positioning for
a Dorsoproximal-45°-Palmarodistal Oblique (DP-45-PDO) view. (D) In this DP-45-PDO view, pathologies
include osteoarthritis of the distal interphalangeal joints (*), rotation of the lateral claw, and osteolysis of
the second phalangeal bones near the joint surface. (E) Positioning for the Horizontal Dorsopalmar (HD-P)
view. (F) In this HD-P view, lesions include marked hoof overgrowth with debris in the sole, osteoarthritis
(*), and an asymmetric joint space associated with collateral ligament injuries (←).
CHAPTER 88 Lameness Diagnosis and Management in Zoo Giraffe 625
A B
C D
E F
626 SE C T I O N 18 Ruminants
TABLE
88.1 Formulary for Giraffe (Giraffa spp.) Lameness Management*
Drug or Possible
Category Treatment Dose Route Dosing Applications Comments
NSAIDs Firocoxib 0.1–0.3 mg/kg p.o. q24h Arthritis
P3 fracture
Laminitis
Swelling
Flunixin 1.1 mg/kg s.c. q24h for Arthritis
meglumine 1–3 days P3 fracture
Laminitis
Swelling
Meloxicam 0.2 mg/kg s.c. once Arthritis
0.3–0.5 mg/kg p.o q24h P3 fracture
Laminitis
Swelling
Phenylbutazone 2–6 mg/kg p.o. q24h Arthritis Recommend
7–8 mg/kg p.o. q48h P3 fracture washout period
Laminitis after 7–14 days
Swelling of daily dosing
Opioids Tramadol 0.5 mg/kg p.o. q24h Arthritis
P3 fracture
Laminitis
Swelling
Joint Adequan 0.75 mg/kg i.m. Loading dose: q5 Arthritis
Supplements days × 7 dose,
then q30 days
Hyaluronan Loading dose: p.o. Loading dose: Arthritis
0.3–0.5 mg/kg q24h × 14
Maintenance dose: days, then
0.15–0.3 mg/kg maintenance
dose q24h
Topicals Cold water 5–10 min Topical q24h—b.i.d. Laminitis Based on
hydrotherapy individual giraffe
temperament,
consider
irrigating with
hose or a foot
bath
DMSO Topical q24h Inflammation Wear gloves when
applying
Epsom salts Foot bath q24h Abscess
Sole foreign body
Antibiotics Ceftiofur 6–9 mg/kg i.m. or q24h 3–14 days Infection n = 1 (calf)
s.c.
Doxycycline 4–5 mg/kg p.o. q24h Infection Sometimes used
in combination
with rifampin
Enrofloxacin 4–8 mg/kg p.o. q24h × 5 days Infection
Sulfadiazine 30–45 mg/kg p.o. q24h Infection For some
trimethoprim individuals, oral
dosing is easier
with powdered
instead of pill
formulations
Tulathromycin 2.5 mg/kg s.c. q72h Infection n=1
Rifampin 6–7 mg/kg p.o. q24h Infection Sometimes used
in combination
with doxycycline
Continued
628 SE C T I O N 18 Ruminants
TABLE
88.1 Formulary for Giraffe (Giraffa spp.) Lameness Management—cont’d
Drug or Possible
Category Treatment Dose Route Dosing Applications Comments
Others Chiropractic Abnormal May help correct
conformation changes
Chronic lameness to spine or
other limbs
associated with
abnormal weight
distribution
Gabapentin 1–5 mg/kg s.i.d. q24h—b.i.d. Suspect spinal Consider starting
injury at lower doses
Chronic lameness and tapering up
as needed. Has
been given with
NSAIDs and
tramadol
Methocarbamol 12–25 mg/kg p.o. q24h Spinal injury
Abnormal body
posture
Therapy laser 3000 J/digit Topical q72h for 1 mo, Arthritis Thermography
8000 J to lateral Topical then 1× Cellulitis may help identify
hock weekly inflammation
q72h for 1 mo and quantify
if treatment
is helping. If
treating digit,
target from
coronary band
to proximal to
fetlock joint
(avoid hoof wall).
Consider using
noncontact
laser on PVC
extension
*Listed doses are anecdotal and based on the author’s clinical experience, as no pharmacokinetic data are currently available for giraffe. Clinicians are advised to
assess whether any of these doses are appropriate to an individual case and should be especially cautious when giving oral medications to ruminants. Results
from an analgesia survey27 or from the Species360 (2017), zims.Species360.org Global Drug Usage reports may also provide additional medicating options.
other cases, clinical improvement has been seen with just Operant Training
corrective hoof trims (Dadone, unpublished results).
A distal metatarsal fracture was successfully managed in Trained medical behaviors using positive reinforcement
a 15-month-old giraffe by using external coaptation with a strategies are key for improving giraffe care.10 This increases
cast, prolonged stall rest, and pain management.26 In this patient access for medical workup and repeat treatments.
case, serial radiographs were used to monitor callus forma- Written safety protocols and restricted contact setups are
tion and bone remodeling, and the final cast was removed strongly recommended to help ensure both human and
5 months after the initial injury. patient safety.10 The author recommends that target training
Surgical management has been reported to be effective in and routine hoof care start for giraffe by about 1 year of
a small number of cases, generally involving young animals. age, similar to what is recommended for domestic horses.
In a 4-month-old giraffe, arthroscopy was used to remove
an avulsion fracture along the lateral trochlear ridge of Prevention
the femur.8 Arthroscopy was also successful for diagnosing
and surgically repairing an osteochondral fragment in the Further study is needed to identify best practices to
metacarpophalangeal joint of an 8-month-old giraffe.25 prevent many causes of lameness in giraffe. Like other
CHAPTER 88 Lameness Diagnosis and Management in Zoo Giraffe 629
megavertebrates, increased attention to foot health, sub- 11. Hummel J, Zimmerman W, Langenhorst T, et al: Giraffe hus-
strate, diet, and routine foot care will likely reduce morbidity bandry and feeding practices in Europe, results of an EEP Survey.
and early mortality in this species. Opportunistic complete In Proceedings. 6th Congress of the European Assoc Zoo Wildl Vet,
2006, pp 71–74.
necropsies that include the joints and feet will help improve
12. Bloch RA, Stephens L, Haefele H: Shoe application and its effects
our understanding of causes of lameness in giraffe. For the on resolution of hoof problems in a giraffe (Giraffa cameloparda-
giraffe in our care, training may provide opportunities to lis). In Proceedings. Annu Conference Am Assoc Zoo Vet, 2013, pp
develop new, more effective treatments for lameness. 44–45.
13. Weidner E, Holland J, Trupkiewics J, et al: Severe laminitis in
Acknowledgments multiple zoo species, Vet Quarterly 34:22–28, 2014.
14. Bush M: Giraffidae. In Miller RE, Fowler ME, editors: Zoo and
Thanks to Amy Schilz; Steve Foxworth; and Drs. Matt wild animal medicine (vol 5), St. Louis, 2003, Saunders, pp
Johnston, Eric Klaphake, Ellen Wiedner, Sushan Han, 625–633.
Myra Barrett, and Julian Fennessy for contributions to this 15. Ihms EA, Rivas A, Bronson E, et al: Pigmented villonodular
chapter. synovitis in a reticulated giraffe (Giraffa camelopardalis), J Zoo
Wildl Med 48:573–577, 2017.
16. Olds JE, Burrough ER, Wilberts BL, et al: Giraffe (Giraffa
References camelopardalis reticulata) arthropathy: post-mortem computed
tomography, gross pathology and histopathologic findings. In
1. Benbow GM, Lyon DG: Experiences with restraint and immo- Proceedings. Annu Conference Am Assoc Zoo Vet, 2015, p 121.
bilization of captive giraffe (Giraffa camelopardalis) in zoos and 17. Zuba JR: Hoof disorders in nondomestic artiodactylids. In
safari parks in Europe. 1997. As cited in Giraffe Husbandry Fowler ME, Miller RE, editors: Zoo and wild animal medicine,
Manual 2003. current therapy (vol 7), St. Louis, 2012, Elsevier Saunders, pp
2. Gage LJ: Medical and husbandry risk management in giraffes: 619–627.
updates on improving giraffe welfare. In Proceedings. Annu 18. Mainka SA, Cooper RM: Hypothyroidism in a reticulated giraffe
Conference Am Assoc Zoo Vet, 2013, pp 140–141. (Giraffa camelopardalis reticulata), J Zoo Wildl Med 20:217–220,
3. Castro AA, Alcantar BE, Stieger-Vanegas S, et al: Anatomic 1989.
description and pathologic findings of chronic foot rot with 19. Hammond EE, Miller CA, Sneed L, et al: Mycoplasma-associated
severe secondary osteomyelitis in a Rothschild’s giraffe (Giraffa polyarthritis in a reticulated giraffe, J Wildl Dis 39:233–237,
camelopardalis rothschildi). In Proceedings. Annu Conference Am 2003.
Assoc Zoo Vet, 2015, pp 156–157. 20. Haschke G, Szentiks CA, Galateanu G, et al: Clinical, anatomic
4. Galateanu G, Göritz F, Szentiks CA, et al: Diagnostic imaging pathological, thermographic and radiological findings in a giraffe
of normal anatomy and pathology in giraffe’s distal limb. In afflicted with arthritis, Der Zoologische Garten 82:259–266, 2013.
Proceedings. Int Conf Dis Zoo Wild Anim, 2013. 21. Basu C, Stoll AL, Dixon J, et al: Osteochondrosis in the distal
5. Dadone L, Olea-Popelka F, Stout E, et al: A wake-up call: femurs of an adult retaliated giraffe (Giraffa camelopardalis reticu-
radiographic evidence of front foot fractures and osteoarthritis lata): macroscopic, radiologic and histologic findings, J Zoo Wildl
in relatively young reticulated giraffe (Giraffa camelopardalis Med 47:359–363, 2016.
reticulata). In Proceedings. Annu Conference Am Assoc Zoo Vet, 22. Shorrocks B: The giraffe: biology, ecology, evolution and behaviour,
2016, p 130. Chichester, 2016, John Wiley & Sons, pp 76–99.
6. deMaar TW, Stewart P, Rosenstein DS: Thermography-assisted 23. James SB, Koss K, Harper J, et al: Diagnosis and treatment of a
diagnosis of a distal phalanx fracture in a reticulated giraffe fractured third phalanx in a Masai giraffe (Giraffa camelopardalis
(Giraffa camelopardalis reticulata). In Proceedings. Annu Confer- tippelskirchi), J Zoo Wildl Med 31:400–403, 2000.
ence Am Assoc Zoo Vet, 2006, p 268. 24. Wakeman K, Sanchez C, Lung N, et al: The use of magnetic
7. Eriksen E: Fracture of the pedal bone in a giraffe, a casuistic resonance imaging to better define hoof pathology in the reticu-
report, Nord Vet Med 24:440–445, 1972. lated giraffe (Giraffa camelopardalis retuculata), J Zoo Wildl Med
8. Quesada R, Citino SB, Easley JT, et al: Surgical resolution of 45:668–671, 2014.
an avulsion fracture of the peroneus tertius origin in a giraffe 25. Radcliffe RM, Turner TA, Radcliffe CH, et al: Arthroscopic
(Giraffa camelopardalis retuculata), J Zoo Wildl Med 41:348–350, surgery in a reticulated giraffe (Giraffe camelopardalis retuculata),
2011. J Zoo Wildl Med 30:416–420, 1990.
9. Bertelsen MF: Giraffidae. In Miller RE, Fowler ME, editors: Zoo 26. Snyder SB, Richard MJ, Hanosh TJ, et al: Management of a
and wild animal medicine (vol 8), St. Louis, 2015, Elsevier, pp distal metatarsal fracture in a giraffe. In Proceedings. Annu Confer-
602–610. ence Am Assoc Zoo Vet, 1997, pp 93–95.
10. Dadone L, Schilz A, Friedman S, et al: Training giraffe (Giraffa 27. Boothe M, Kottwitz J, Harmon R, et al: Results of the megaver-
camelopardalis reticulata) for front foot radiographs & hoof care, tebrate analgesia survey: giraffe and hippopotamus, J Zoo Wildl
Zoo Biol 35:228–236, 2016. Med 47:1049–1056, 2016.
89
Mass Mortality Events Affecting
Saiga Antelope of Central Asia
RICHARD ANTHONY KOCK AND SARAH ROBINSON
S
aiga (Saiga tatarica) are a unique Central Asian ante- aggregations over a week or so and produce one to three
lope, composed of two subspecies (S. t. tatarica and calves each, with the majority of females reproducing in
S. t. mongolica), relatively unchanged in form since their first year of life and having the highest relative fetal
the mid to late quaternary period based on fossil remains.1 biomass of any mammal.9 This extraordinary lifestyle and
It is a species highly adapted to the semiarid desert steppes reproduction is not an accident. These harsh lands have
of Russia, Kazakhstan, and Mongolia with more than 90% developed a hard bargain with this species; annual losses
of the global population in the Uralsk and Betpak-Dala will be naturally high, with sometimes catastrophic declines,
regions of Kazakhstan. They are ruminants, exploiting a from which they can and do rapidly recover.
rich diversity of plants for their nutrition2–6 in highly saline, Their biggest challenge in recent times has come from
often mineral (e.g., copper) deficient pasture and sometimes humans, an even more daunting challenge than the envi-
soil toxic conditions with high boron that few species can ronment and from which they can only run. Poaching with
tolerate.7 Temperatures plummet to −16°C on average in high velocity rifles has been the single most serious cause of
January, with severe conditions reducing this further to premature death over recent decades, with numbers at the
−40°C at times, and temperatures rise to higher than 30°C end of the 20th century declining dramatically from more
in summer. To cope with these harsh conditions, saiga have than a million to a few tens of thousands of individuals.10
a compact body mass and grow a highly insulated thick hair As with so many other species, this toxic mix of human
coat, shed in spring. They possess unusual nasal anatomy persecution and increasing environmental challenges
with an extended proboscis, which reduces the risks associ- such as habitat fragmentation threatens this species with
ated with breathing the freezing and dusty air and may help extinction.11
drying of exhaled air to help conserve precious moisture The major causes of mass mortality events (MMEs),
that is unavailable over much of the year. Key to survival other than humans, are heavy snowfall or freezing rain
in these semidesert conditions is annual migration1,2 and (known as dzhut in Kazakh) and disease-related events
careful calving site selection to optimize survival.8 The (Table 89.1). The majority of diseases were attributed to
antelope runs at remarkable speeds for prolonged periods Pasteurella species by contemporary observers and described
and covers huge distances over a year or even a day, avoid- under the umbrella term of “pasteurellosis.” The largest of
ing contact with humans consistent with historic poaching these occurred in 1981,12,13 1984,14,15 1988,16–18 and 2015,19
pressure. With few other prey species in these vast lands, causing the death of tens to hundreds of thousands of
predators seek out saiga, but none other than humans in animals (Table 89.1; Fig. 89.1). Some smaller disease events
4 × 4 vehicles can keep up. Predators benefit from chance have also been attributed to Pasteurella in 1974,20 2011,21,22
encounters and two periods of the year when the saiga 2012,23 and 2013.24 Both large and small events were inad-
must stop for calving and rutting. These periods are highly equately documented for diagnosis, with the exception of
synchronized and tuned into the vegetation cycles to ensure 2015, and it was notable in 1974 that the saiga had been
a rising plane of nutrition in spring for calving and lactation chased off feed crops over a number of days prior to their
and optimal body condition in the autumn for the rut to death. The majority of Pasteurella-related die-offs occurred
ensure maximum fertility. The population of females and around calving but not exclusively, and a number of the
some younger males cluster around May in herds of a few largest events were clustered in the northern part of the
to tens of thousands of animals. They form dense birthing Betpak-Dala population range (Fig. 89.2).
630
CHAPTER 89 Mass Mortality Events Affecting Saiga Antelope of Central Asia 631
TABLE
89.1 A Summary of Mass Mortality Events in Saiga (Saiga tatarica) Affecting More Than 1000 Animals
Of the largest MMEs, in 1981 “pasteurellosis” was Die-off is not unique to Kazakh populations. In late
given by the cited sources as the cause of death, but the 2016, the small Mongolian population in the Gobi Altai
specific identity of the organism and detailed gross pathol- region of Western Mongolia became infected with peste
ogy information that might have allowed confirmation of des petits ruminants virus (PPRV), as a result of spillover
the syndrome were not available. P. multocida was isolated from livestock during the first epidemic of this disease
from dead saiga in 1988, and there is reasonable supporting to be recorded in the country.30 The saiga proved to be
evidence for hemorrhagic septicemia (HS), but animals had highly susceptible to the virus, perhaps indicative of
suffered from severe dhzut during the previous winter. In their close phylogeny to the family Caprinae (the main
1984, an organism described as P. haemolytica was isolated, host to this disease), in which they used to be classified,
and the die-off was possibly related to a pneumonic form while only more recently being incorporated into the
of pasteurellosis. In the 2010 die-off in Ural, Pasteurella Antilopini on a basis of modern genetic analysis. In the
multicoda was isolated, but the symptoms resembled those Altai the saiga expressed a virulent form of the disease
of a pasture-related disease (e.g., bloat or fog-fever), more with mortality reaching more than 5000 animals or 54%
than those of Pasteurella-related syndromes. A small die-off of the population within a matter of weeks,31,32 and the
of 400 animals among survivors in 2011, again with epidemic continued up until June 2017, affecting the
isolation of the same organism, occurred at exactly the entire population and other wildlife species. In addition,
same location as the previous year’s die-off, while animals much was written on foot and mouth disease (FMD)
grazing elsewhere remained healthy, suggesting location and in saiga following outbreaks in the 1950s and 1960s,
perhaps pasture as the primary factor.25–27 but these tended to cause mortality mainly in calves,
For the 2015 event, the identity of the principal patho- and only on two occasions were many thousands of
gen, P. multocida serotype B, and syndrome, HS, is of little animals affected.2 Besides these reported infections, and
doubt.19 Moreover, climatic conditions during this event a number of other MMEs with uncertain diagnoses and
and the two other likely large HS die-offs in Betpak-Dala etiologies, saiga die from expected mortality in calves
(1981, 1988) were found to be on average warmer and more around time of birth, from sudden weather shifts, from
humid compared with control calving areas in which die- low birth weight and weakness where there is failure to
offs did not occur.19 Such patterns have also been associated suckle, from predation during the aggregation, and over
with HS prevalence in domestic livestock.28,29 the first few months of life. In adults, mortality occurs
632 SE C T I O N 18 Ruminants
• Figure 89.1 Locations of Saiga (Saiga tatarica) die-off events in Betpak-Dala 1974–2015. Notes
to Fig. 89.1: 2015 sites: Sources of estimated deaths: authors’ observations; Zhubaniyz.40 2012 site:
Zuther.23 1988 sites: The figures given here, originally based on total deaths of over 400,000, which are
believed to have been an overestimate, have been re-scaled proportionally to give the eventual estimated
total of 270,000.16 They are thus indicative only and meant to give an impression of relative numbers dying
at each site. 1981 site: Institute of Zoology and Department of Reserves and Hunting.12
Discussion
Saiga antelope remain a keystone species of the Central
Asian steppe which, following extinctions of the wild
horse and ass, lacks significant competition for the dry
desert-like grasslands other than with livestock and
rodents. Their populations have been sorely depleted, due
• Figure 89.2 Mass mortality of more than 60,000 saiga (Saiga
to displacement by agriculture and settlement in parts
tatarica) in Amangeldy, Turgai River basin, Kazakh steppe in May
2015, showing all the adults in the picture dead and calves wandering
of their range, and more recently from disease causing
lost, apparently unaffected by this catastrophe. MMEs and poaching. Older literature is remarkably silent
about MME other than from dzhut and FMD events
in the 1950s and 1960s, which affected up to 50,000
calves.2,36 Pasteurellaceae are prominent as a feature in
recent events, but this may reflect modern diagnostic
CHAPTER 89 Mass Mortality Events Affecting Saiga Antelope of Central Asia 633
mestakh okota saigakov Betpakdalinskoi populyatsii v 2013g]. [Vestnik Sel’skokhozyaistvennoi Nauki Kazakhstana] 5:66–67,
Astana, 2013. 1980.
6. ACBK: Report on calving sites of the Betpak-dala saiga population 21. Kock R, Grachev Y, Zhakypbayev A, et al: A retrospective
in 2014 [Otchet po issledovaniyam na mestakh okota saigakov assessment of saiga antelope Saiga tatarica die-off in Western
Betpakdalinskoi populyatsii v 2014g]. Astana, 2014. Kazakhstan 2010-2011, Saiga News 1–4, 2014.
7. Alikaev VA: Recent work on mineral nutrition of farm animals 22. Kock R, Rycroft A: Rapid response to the Saiga antelope emer-
[Novoe v uchenii o mineral’nom pitanii sel’skokhozyaistvennykh gency in Western Kazakhstan: Fauna and Flora International,
zhivotnykh], Veterinariya 35:109–117, 1958. 2012.
8. Singh NJ, Grachev IA, Bekenov AB, et al: Saiga antelope calving 23. Zuther S: Mass death in the Betpak-dala saiga population, Saiga
site selection is increasingly driven by human disturbance, Biol News 15:4, 2012.
Conserv 143:1770–1779, 2010. 24. Novosti Kazakhstana: Reason for mass dieoff of Saiga was
9. Kühl A, Mysterud A, Erdnenov GI, et al: The ‘big spenders’ of pasteurellosis [prichinoi massovoi gibeli saigakov stal pasterellez]
the steppe: sex-specific maternal allocation and twinning in the Novosti Kazakhstana/Namba Media, 2013.
saiga antelope, Proceedings of the Royal Society B Biological Sciences 25. Pinion V, Kock R: Is pasture structure and plant-composition
274:1293–1299, 2007. the key to explaining the mass die-off of the Saiga antelope,
10. Milner-Gulland EJ, Kholodova MV, Bekenov A, et al: Dramatic Saiga tatarica tatarica, in the Borsy region of Kazakhstan?: Royal
declines in saiga antelope populations, Oryx 35:340–345, 2001. Veterinary College London, 2012.
11. IUCN/SSC Antelope Specialist Group & Saiga Conservation 26. Dieterich T, Sarsenova B: Examination of the forage basis of saiga
Alliance: Overview Report on Conservation Status and MOU in the Ural population on the background of the mass death in
Implementation: Convention on Migratory Species, 2015. May 2010 and 2011. Material from the International Scientific
12. Institute of Zoology and Department of Reserves and Hunting: Conference ‘Biological Diversity of the Asian Steppes’, 2012.
Report on the aerial counts of saiga in Kazakhstan from 23 March 27. Dancer A, Pinion V, Kock R, et al: Remote and in situ analyses
to 28 April 1981 [Ochet po provedeniu aviavizual’nogo ucheta of the potential causes of saiga antelope die-offs in Western
saigakov v Kazakhstane c 23 marta po 28 aprelya 1981], Alma-Ata, Kazakhstan, Saiga News 16:16–18, 2013.
1981, Council of Ministers and Academy of Sciences of the 28. Lunitsyn VG, Bakulov IA: Pasteurellosis in deer [Pasterellez
Kazakh SSR. pantovykh olenei], Veterinaria 6:37–40, 1985.
13. Institute of Zoology and Department of Reserves and Hunting: 29. Dutta J, Rathore BS, Mullick SG, et al: Epidemiological studies
Report of the regular multidisciplinary Saiga monitoring expedition on occurrence of haemorrhagic septicaemia in India, Indian Vet
1981, Alma-Ata [Ochet postoyannoi kompleksnoi ekspeditsii J 67:893–899, 1990.
po saigaku 1981], 1982, Council of Ministers and Academy of 30. OIE FAO CMC: Rapid assessment for the control and pre-
Sciences of the Kazakh SSR. vention of further spread of peste des petits ruminants (PPR).
14. Institute of Zoology and Department of Reserves and Hunting: Mission Report FAO, Rome, 2016, p 22.
Report on the aerial counts of saiga in Kazakhstan from 4 April 31. OIE FAO CMC: Investigation of Peste des Petits Ruminants
to 1 May 1984 [Ochet po provedeniu aviavizual’nogo ucheta (PPR) among wild animals and its potential impact on the
saigakov v Kazakhstane c 4 aprelya po 1 maya 1984], Alma-Ata, current PPR situation in livestock. FAO, Rome, 2017;29.
1984, Council of Ministers and Academy of Sciences of the 32. WWF: More than 4000 Mongolian saigas die in a disease
Kazakh SSR. outbreak. [wwf.panda.org], 2017.
15. Aikimbaev MA, Martinevski IL, Altukhov AA, et al: On 33. Barros Sa Braz D: Background mortality of Saiga antelope
the analysis of the pathogenic Pasteurella from Saiga during (Saiga t. tatarica) during calving season in Kazakhstan: Univer-
February-March 1984 in the Ural region [O sluchayak vydeleniya sidadmestrado Integrado EM Medicina Veterinariae de Lisboa,
pasterelleza ot saigakov v fevrale-marte 1984 goda v Uralkskoi Lisbon, Portugal, 2016.
oblasti], Seriya Biologicheskaya 4:39–41, 1985. 34. Bayarbaatar B: Factors affecting survival and cause-specific
16. Institute of Zoology and Betpak-dala State Hunting Organisa- mortality of saiga calves (Saiga tatarica mongolica) in Mongolia.
tion: Aerial-Visual Report on the Betpak-dala-Arisskoi Populations Department of Environmental Conservation: University of Mas-
of Saiga in Kazakhstan [Aviavizual’nogo uchet betpakdalinsko- sachusetts Amherst, USA, 2011, p 62.
arysskoi grupirovki saigakov v Kazakhstane], Alma-Ata, 1988, 35. Sánchez-Monge F: Calving status and commensalism of Pas-
Academy of Sciences of the Kazakh SSR. teurella multocida in the surviving Betpak-dala saiga population
17. Institute of Zoology and Betpak-dala State Hunting Organisa- in May 2016, after a mass mortality event in May 2015: Royal
tion: Report of the regular Multidisciplinary Saiga Monitoring Veterinary College, University of London, UK, 2016.
Expedition 1988 [Ochet postoyannoi kompleksnoi ekspeditsii po 36. Bannikov AG, Zhirnov LV, Lebedeva LS, et al: The biology of the
saigakam 1988], Alma-Ata, 1989, Academy of Sciences of the saiga [biologiia saigaka], Moscow, 1961, USSR.
Kazakh SSR. 37. Rotshild EV: Infections in nature. Dangerous diseases as viewed by
18. Turgai Regional Executive Committee: On Massive Mortality of a naturalist [Opasnye nedugi glazami naturalista], Environmental
Saiga in Turgai Oblast of the Kazakh SSR [O Sluchae Massovogo Epidemiology [Envayronmental’naya epidemiologiya] 5:434–740,
Padezha Saigakov v Turgaiskoi Oblasti Kazakhskoi SSR], 1988. 2011.
19. Kock R, Orynbayev M, Robinson S, et al: Saigas on the brink: 38. Rotshild EV, Yevdokimova AK, Amgalan Z: Abnormalities of
multi-disciplinary analysis of the factors influencing a mass mineral composition in plants as a factor in the loss of the Mon-
die-off event, Science Advances 4(1), 2018. golian gazelle in Mongolia [Anomalii mikroelementogo sostava
20. Statsenko NI: Cases of pasteurellosis in saiga in Kurgal’dinskii rastenii kak faktor padezha dzherenov v Mongolii], Bulyetin
nature reserve [Sluchai pasterelleza Saigakov v Kurgal’dzhinskom Moskovskovo O-va ispytatelei prirody Otdel Biologiyi 93:35–42,
zapovednike], Journal of Agricultural Sciences of Kazakhstan 1988.
CHAPTER 89 Mass Mortality Events Affecting Saiga Antelope of Central Asia 635
39. Berkinbaev O: A study of diseases among Saiga in Kazakhstan 42. Kock R: Final Report of Mission by Prof Richard Kock to
[Izuchenie boleznei Saigakov v Kazakhstane], Diseases and Kazakhstan September 13th to October 1st 2011 and related
Parasites in Wild Animals [Bolezni i Parazity Dikikh Zhivotnykh] analysis: Royal Veterinary College, 2011.
32–41, 1992. Moscow, Russia. 43. Institute of Zoology and Department of Reserves and Hunting:
40. Zhubaniyz M: Saiga in the Irgiz-Turgai State Nature Reserve Report of the regular multidisciplinary saiga monitoring expedi-
Technical Workshop for Saiga Antelope Experts and Third tion [Ochet postoyannoi kompleksnoi ekspeditsii po saigaku 1984],
Meeting of the Signatories to the Memorandum of Understand- Alma-Ata, 1985, Council of Ministers and Acadamy of Sciences
ing concerning Conservation, Restoration and Sustainable Use of the Kazakh SSR.
of the Saiga Antelope. Tashkent, 2015.
41. Dieterich T: Saiga mass death turgay - betpak dala population
may 2012 preliminary botanical investigations, Astana, 2012,
ACBK.
90
Musk Ox Sedation and Anesthesia
CARSTEN GRØNDAHL
M
usk oxen (Ovibos moschatus) are some of the precipitation in the arctic in May, and calves may develop
toughest ruminants—their resilience and toler- pneumonia and die if they become hypothermic from a
ance to environmental challenges in the Arctic soaking rain. The calves are also very sensitive to overheat-
are unsurpassed. They are examples of extreme adaptation. ing. Great care should be taken on sunny days during the
Musk oxen are true ruminants and, therefore, when sedated first months of life, as calves may fatally overheat, even
or anesthetized, must remain in a sternal position to allow when environmental temperatures are not elevated. Ensur-
for saliva to drain and for rumen gases to be eructated. This ing shade and monitoring the cow and calf to ensure the
minimizes the workload of ventilation and maximizes gas use of shade provided is crucial. It is important not to put
exchange. Their lungs are small, and supplemental oxygen calves in with other musk ox cows other than their dams,
is almost always mandatory, as the arterial oxygenation as cows will not tolerate other calves and will often show
drops rapidly when oxygen is not supplemented in most aggression toward the unfamiliar calf.
anesthetic combinations.1
Restraint/Sedation/Immobilization
Biology
Reasons for restraint include identification and marking of
The musk ox is thought to have experienced significant calves, treatment of calf diarrhea, transport, health surveil-
genetic bottlenecks. Despite these bottlenecks, two sub- lance, hoof trimming, treatment of trauma from other musk
species of musk ox, O. m. wardi (white faced musk ox ox, dystocia, and blunting of horn tips with epoxy.
or Greenlandic musk ox) and O. m. moschatus (Barren A special concern regarding restraint/anesthesia is that
Ground musk ox), have been commonly accepted based on musk oxen are very heat sensitive. Do not restrain/sedate
their morphologic differences and geographic separation. or immobilize musk oxen of any age on warm days, and
However, when control-region sequences of mitochondrial continuously monitor temperature with a rectal probe.
DNA were compared among 37 musk oxen, there was
little variation found among the musk oxen sampled. These Physical and Chemical Restraint
results do not support musk oxen subspecies.2
In the first 3 months, physical restraint may be performed
Distribution, Habitat, Size, and Weight with experienced “cowboys.” We often use a bale of straw
as a treatment table, and if physical restraint is performed
Musk oxen primarily live in the Canadian Arctic and frequently, the calves may become tame (Fig. 90.1).
Greenland, with small introduced populations in Sweden, If chemical restraint is needed for these young calves (such
Siberia, Norway, and Alaska. The size varies but is often as for transport to a medical facility, radiography, ultrasound,
overestimated due to the thick fur. The Canadian variant or other procedure), sedation using an alpha-2 agonist opioid,
is often bigger (weighing 286 kg on average and up to and benzodiazepine combination is usually adequate. For
410 kg3) than the Greenlandic type, the latter having free- example, a 3.5-month-old calf (weighing 22 kg) was sedated
ranging cows weighing 160–210 kg and free-ranging bulls for transport and radiology using 1 mg detomidine, 2 mg
weighing 270–320 kg.4 butorphanol, and 1 mg midazolam. The calf was calm and
recumbent but awake during transport and radiography.
Environmental Considerations Larger calves older than 3 months up to 1 year can be
sedated with an alpha-2 agonist, butorphanol/methadone,
Calves are born on snow-covered ground in May and should and midazolam, which may be supplemented with low dose
not be on gravel substrate, as they will often eat the gravel ketamine.
and develop geosediment-related disease. They do not have For animals over 1 year, sedation is recommended only
water-shedding fur. It is important to note that there is no in very tame or debilitated animals, as sedation of healthy
636
CHAPTER 90 Musk Ox Sedation and Anesthesia 637
Immobilization
Many drug combinations have been used in musk ox
including xylazine-ketamine, ketamine-medetomidine,
tiletamine-zolazepam-medetomidine, or ketamine-medeto-
midine-butorphanol. Combinations with potent opioids
include carfentanil-xylazine, carfentanil-xylazine-ketamine,
etorphine-xylazine, or etorphine-xylazine-midazolam-
ketamine. See Table 90.1 for Zoological Information
Management System (ZIMS) (Species360 [2017], zims.
Species360.org) anesthesia summary reports doses used
(primarily North American use). Other combinations
like BAM (butorphanol-azaperone-medetomidine) or tile- • Figure 90.2 Mask inducing a young musk ox (Ovibos moschatus)
tamine-zolazepam combinations have been used by col- calf with sevoflurane in oxygen.
leagues, but published evaluations are not available
(Kimberlee Beckmen, personal communication). If walking into a transport crate is the desired procedure,
Long needles of adequate length to penetrate the thick administer 75%–80% of a normal dose, and after approxi-
hair coat should always be used. Snow and ice can coat mately 6–8 minutes, blindfold the animal and guide it into
the hair making intramuscular (IM) darting challenging. the crate.
Lubricate the needle with sterile silicone oil (not spray).
If there is not an adequate effect after 12–16 minutes, Gas Anesthesia
dart again with 50%–100% of initial dosing depending For prolonged procedures, gas anesthesia is an option, and
on the efficacy of the first dart. An animal not recumbent both sevoflurane and isoflurane work well. When mask
would receive a full dose, a heavily sedated animal that inducing smaller calves or sedated juveniles, sevoflurane
is recumbent but able to get up when approached would gives less mucosal membrane stimulation and a faster and
receive half the initial dose, and a recumbent animal that easier induction of anesthesia (Fig. 90.2). Intubating musk
cannot get up when approached, but who is able to lift the ox is as challenging as for all small- to medium-sized rumi-
head and look around, would receive 20%–30% of initial nants. The largest animals may be intubated manually with
dosing often by hand injection, ensuring that a good IM a guide tube through the endotracheal tube and by ensuring
injection is made. the guide tube placement is between the plica vocalis. Most
638 SE C T I O N 18 Ruminants
TABLE
90.2 Author’s Recommended Doses for Sedation and Anesthesia in Musk Ox (Ovibos moschatus)
Musk Ox Sedation
Musk Ox Anesthesia
Age Drug combination (total mg/animal) or per 100 kg bodyweight (bwt)
Yearling 0.6–0.8 mg etorphine + 6–8 mg xylazine + 8–10 ketamine + 1 mg
midazolam
Subadult 14–18 months 0.8–1.0 mg etorphine + 8–10 mg xylazine + 8–10 mg ketamine +
1–2 mg midazolam
2 years old 1–1.2 mg etorphine + 10–12 mg xylazine + 10–12 mg ketamine +
1–2 mg midazolam
Adult cows (per 100 kg bwt) 0.8–1 mg etorphine + 8–10 mg xylazine + 10 mg ketamine + 1 mg
midazolam
Adult bulls (per 100 kg bwt) 0.6–1 mg etorphine + 6–10 mg xylazine + 10 mg ketamine + 1 mg
midazolam
Bulls usually need less per kg bodyweight than cows: 1.8 mg etorphine + 18 mg xylazine + 10 mg ketamine (used
several times bulls (300 kg) have been anesthetized successfully on a docile 315 kg bull for a hood trim lasting 45 min)
with a dose calculated for 200 kg cow
animals will require a long straight laryngoscope with a good Valby, Denmark) are effective at a dose of 1 mL/80–100 kg
light source for direct visualization of the larynx and place- for the introduction of new herd members, introduction
ment of the tube. Use preferably high-volume low-pressure to a new enclosure, or transport. Both the short-acting (72
cuffs. If using a low volume high-pressure cuff, take special hours) and depot (21 days) formulations are very effec-
care not to overinflate the cuff, which can lead to tracheal tive. Older reports6 indicate a good effect and negligible
mucosa ischemia and serious complications. See Table 90.2 side effects using perphenazine (Trilafon Dekanoat, Orion
for recommended doses of sedation and anesthesia. Pharma, Denmark) at a dose of 0.51 mg/kg.
Free-ranging musk ox immobilization (e.g., satellite
collar) involves slightly higher doses; Greenlandic musk Perianesthesia
oxen cows in October,5 body weight around 200 kg, have
successfully been immobilized with 1–1.2 mg etorphine/ If possible prior to anesthesia, food and water are withheld
100 kg + 15 mg xylazine/100 kg + 20 mg ketamine overnight for adults; however, water is not withheld from
/100 kg + 0.15 mg medetomidine /100 kg. juvenile animals.
It is the author’s experience that combining two dif- During the anesthetic procedure, always keep recumbent
ferent alpha-2 agonists (xylazine and medetomidine) and animals in a sternal position or as close as possible to sternal.
two opioids (butorphanol and methadone) gives a synergic Point the nose downward so saliva may dribble out.
effect while not increasing the side effects. This is maybe due Supplement oxygen by placing a nasal insufflation
to different subtype affinity for the four alpha-2 receptors catheter deep into the nasal cavity (1–2 L O2/100 kg) and
and the mu and kappa opioid subtype receptors. monitor arterial saturation using pulse oximetry.
When working with adult animals, secure the workspace.
Use of Long-Acting Tranquilizers Have a designated person responsible for the head and apply
Long-acting tranquilizers such as zuclopenthixol-acetate rubber stoppers on the horn tips (Fig. 90.3). The person at
Cisordinol-Acutard 50 mg/mL and zuclopenthixol dec- the head should also monitor eye and ear reflexes together
anoate Cisordinol depot 200 mg/mL (Lundbeck Pharma, with ventilation and be able to manually restrain the head
CHAPTER 90 Musk Ox Sedation and Anesthesia 639
• Figure 90.3 Protecting people by placing rubber stoppers on the • Figure 90.4 Drawing blood from the metacarpal vein of a musk ox
musk ox (Ovibos moschatus) horn tips. A short tube is inserted in one (Ovibos moschatus). The head is to the right.
nostril for end tidal CO2 measurements.
by holding the horns as needed. Strapping the legs together clipped skin (inside hind leg) and/or venous blood gas
with soft lifting straps with a person holding the end of the measurements (e.g., I-stat).7
loop is a very effective restraining tool.
When performing a hoof trim with power tools, protect Use eye drops (clear type, not petroleum-based, as the
the eyes of the musk ox, as well as the eyes of the people latter will impair vision and cause unnecessary distress
around the musk ox. during the recovery phase).
During anesthesia, it is possible to deepen or lighten Blood collection is possible from the tarsal vein (when the
the depth of anesthesia of the musk ox when using the animal has a summer coat) just above the tarsal-metatarsal
potentiated opioids combined with alpha-2 agonists. joint or the metacarpal vein (Fig. 90.4). Both locations
If the animal is too deep, characterized by respiratory are more easily accessed by using a tourniquet made from
depression and low blood pressures, partially reverse some thick rubber hoses or similar. The jugular vein can also be
of the depression induced by the alpha-2 agonist and used if a larger quantity of blood is needed. The use of
stimulate respiration, by giving a microdose of atipamezole extension tubing on the needle is advantageous, especially
together with doxapram IM (e.g., 200 kg cow receives when a colleague is available to draw the blood from the
1–2 mg atipamezole and 10–15 mg doxapram total dose). tube end.
If the animal is too light (i.e., able to hold the head up Weighing the animal is possible even in remote areas
and move the head around, but not able to get up when by using a tarpaulin and 4–6 ordinary suitcase scales. At a
approached), hand inject 25%–50% of the initial dose. If given time point, the scale weights are added and then the
the animal starts to wake up during a procedure, supple- procedure is repeated—after subtracting the weight of the
ment with ketamine 0.5–1 mg /kg IM. tarpaulin, the weight of the musk ox often differs less than
1% between the measurements.5
Monitoring During Anesthesia There are different opinions regarding the routine use of
Basic: Depth of anesthesia (reflexes) and ventilation (fre- prophylactic antibiotic treatment after immobilization in
quency and depth), saturation (pulse oximetry), and nondomestic ruminants. My policy is not to use antibiot-
continuous rectal temperature. ics when performing a hoof trim or other scheduled tasks
Medium: Basic monitoring supplemented with noninvasive but to use a broad spectrum long-acting antibiotic when
blood pressure measurements (carpal or tarsal cuff place- immobilization for transport (to prevent shipping fever)
ment) and end tidal CO2 (ETCO2, nasal tube connected and when working on nonfasted animals to minimize the
to a sidestream or mainstream capnography (EMMA). I pulmonary reactions if a few drops of ruminal fluid or saliva
cut a normal polyvinyl chloride single use endotracheal are accidently inhaled.
tube in half (internal diameter 8 mm for an adult cow) When reversing potent opioids in animals expected to
and insert it into one nostril (only about 3–4 cm depth) have moderate or severe pain after the procedure, buprenor-
and connect it to the capnography (see Fig. 90.3). phine 0.01–0.02 mg/kg intravenous (IV) or IM can be used
Advanced: Basic and medium monitoring supplemented as the reversing agent. The onset of action is approximately
with arterial blood samples and gas analysis (e.g., I-stat) 20 minutes for full opioid reversal, but significant respira-
and direct arterial blood pressure measurements. Arte- tory improvement is seen after about 5 minutes. There is
rial access is possible by cannulating the auricular artery residual analgesia for probably 6–8 hours after reversal,
(Lian et al.1), and tissue perfusion assessment may be in contrast to animals receiving naltrexone. Naltrexone
done using near infrared spectrophotometry (NIRS) on completely antagonizes the administered opioid, including
640 SE C T I O N 18 Ruminants
C
apripoxviruses (CaPVs) are the cause of diseases in may vary, but the severity of clinical disease often depends
domestic ruminants, having significant economic on the infected species, age, and immune status.3–5 Mor-
impact on the livelihood of farmers in endemic bidity and mortality numbers in wild animals are poorly
areas. The diseases they cause are characterized by fever and recorded and mainly anecdotal. Mortality rates as high as
nodular lesions on the skin and internal organs. The viruses 100% were reported in 16 captive ruminant species in a
are expanding their historical territory and have the poten- GTPV outbreak event in 2015 (Table 91.1).6
tial of becoming emerging disease threats because of global
climate change and increasing trade in animals and animal Range and Host-Specificity
products. A number of wildlife species have been implicated
in the disease epidemiology, but clinical symptoms have There are distinct differences between the host species and
hardly been reported. This chapter addresses the rising the geographic distribution of SPPV, GTPV, and LSDV.
concern for disease in nondomestic ruminants after a severe CaPV are considered highly host specific, but exceptions are
outbreak with high mortality in captive wild animals in Qatar recorded sporadically. Historically they infect only domestic
in 2015. ruminant species and have no zoonotic potential.7 They are
expanding their range and have the potential of becoming
Etiology emerging disease threats.3 The geographic range of SPPV
and GTPV extends from Africa north of the Equator, across
The genus Capripoxvirus is classified in the subfamily Chor- the Middle East and Turkey, to the Indian subcontinent and
dopoxvirinae of the family Poxviridae. It is composed of Asia, including parts of Russia and China. In recent years
three virus species—namely sheeppox virus (SPPV), goatpox the range has extended with disease outbreaks occurring in
virus (GTPV), and lumpy skin disease virus (LSDV).1,2 Vietnam (2005 and 2008), Mongolia (2006 and 2007), and
CaPVs are brick-shaped, enveloped, double-stranded DNA Greece (2008).3,5 Most SPPV and GTPV strains are host
viruses with genomes approximately 150 kbp in size. They specific and cause clinical disease in either domestic sheep
are among the largest viruses, measuring 170–260 nm by or goats, while some strains have equal virulence in both
300–450 nm. The three viruses in the genus are closely species.3 In 2015 an outbreak of GTPV in a wildlife col-
related antigenically and therefore cannot be distinguished lection in Qatar caused clinical disease and high mortality
serologically. They are also difficult to distinguish morpho- in 16 species of caprinae, antilopinae, and hippotraginea
logically from orthopoxviruses.3 All CaPV outbreaks are (see Table 91.1).6 This was the first documented case of
categorized as notifiable diseases in the World Organisation GTPV in wild animals, although similar outbreaks have
for Animal Health (OIE) guidelines. been witnessed in the region (T. Cavero, personal com-
munication, January 16, 2017).
Epidemiology Originally LSDV was restricted to sub-Saharan Africa,
but today it occurs in most African countries (including
Highly contagious SPPV and GTPV may cause very high Madagascar). Since 1990 outbreaks have been reported
morbidity rates (70%–90%) in domestic animals. Mortality in the Middle East and between 2013 and 2015 in Iraq,
may be up to 50% and as high as 100% in young or naive Iran, Turkey, Cyprus, and Greece.3,5,8 In domestic cattle,
animals. For LSDV infections, morbidity rates may vary LSDV causes clinical disease, but susceptibility of wildlife
from 5%–45% and sometimes be up to 100%. Mortality is seen sporadically (Table 91.2). Natural infections have
usually remains below 10%, although mortality rates over been reported in Asian water buffalo (Bubalus bubalis),
75% have been recorded. The virulence of different CaPV springbok (Antidorcas marsupialis), and an Arabian oryx
641
642 SE C T I O N 18 Ruminants
TABLE
91.1 Wild Ruminants Affected in Goatpox Outbreak, AWWP, Qatar, 2015
Affected Species No. of Individuals at Risk No. of Deaths Mortality Rate (%)*
Laristan mouflon (Ovis orientalis laristanica) 48 25 52.08
Iranian wild goat (Capra aegagrus) 39 37 94.87
Nubian ibex (Capra ibex nubiana) 2 2 100.00
Addax (Addax nasomaculatus) 47 38 80.85
Arabian oryx (Oryx leucoryx) 43 6 13.95
Gerenuk (Litocranius walleri) 28 10 35.71
Beira antelope (Dorcatragus megalotis) 2 2 100.00
Soemmering’s gazelle (Gazella soemmeringi) 112 67 59.82
Idmi gazelle (Gazella gazella) 93 8 8.60
Yemeni gazelle (Gazella gazella cora) 40 6 15.00
Erlanger gazelle (Gazella gazella erlangeri) 7 7 100.00
Chinkara (Gazella bennettii) 24 4 16.67
Dama gazelle (Gazella dama ruficollis) 47 3 6.38
Speke gazelle (Gazella spekei) 59 10 16.95
Persian goitered gazelle (Gazella s. subgutturosa) 172 3 1.74
Arabian goitered gazelle (Gazella s. marica) 97 4 4.12
Red-fronted gazelle (Gazella rufifrons) 51 1 1.96
Dorcas gazelle (Gazella dorcas isabella) 77 1 1.30
Pelzeln gazelle (Gazella pelzelni) 41 4 9.76
(Oryx leucoryx), while clinical signs have been demonstrated of cuts and skin abrasions. The amount of viral shed-
in impala (Aepyceros melampus) and giraffe (Giraffa camelo- ding correlates with the severity of clinical disease, and
pardalis) after experimental inoculation with LSDV.9–13 chronically infected carriers do not occur. Due to high virus
However, so far there have not been any confirmed LSDV concentrations in the skin, indirect transmission via insect
outbreaks in any wildlife species. Antibodies against LSDV vectors has been suggested.18
have been detected in a range of African game species. In contrast to SPPV and GTPV, the main path of trans-
Nonetheless, serologic positivity does not necessarily mean mission of LSDV is mechanical via biting insects, and direct
that the virus is being replicated and excreted.11,14–17 The contact is considered a minor source of infection.19,20 The
presence of antibodies in an animal, however, does indicate virus seems to be capable of spreading over long distances,
its susceptibility to CaPV and its potential involvement in depending on factors benefiting the arthropod vectors.5,21
the epidemiology of the disease. Nevertheless, the role of
wildlife in the epidemiology of LSDV is currently not well Clinical Signs and Pathology
understood.16
Clinical symptoms may vary from mild to severe and even
Transmission lead to death, depending on susceptibility of the host and
virulence of the strain. Following infection, rectal tempera-
SPPV and GTPV are highly contagious and may remain ture rises to above 40°C after an incubation period of 8–14
viable in crusts and scabs in the environment for several days. Skin nodules of 1–5 cm in diameter, involving all
months. Shedding occurs via ulcerated papules on the layers of the skin, develop concurrently with the fever and
mucous membranes and nasal, oral, and conjunctival secre- may cover over 50% of the skin surface (Fig. 91.1). Draining
tions. Transmission is usually through aerosols and close lymph nodes are often enlarged, especially the prescapular
contact with infected animals or by indirect contamination lymph nodes. Animals show depression, loss of appetite,
CHAPTER 91 Capripoxviruses in Nondomestic Hoofstock 643
TABLE
91.2 Lumpy Skin Disease Virus Detected in Exotic Ruminants
and are reluctant to move. Ulcerative lesions on mucous bacterial infections and pneumonia are common, and death
membranes of the eyes, nose, mouth, pharynx, and tongue may occur at any stage of the disease. Lesions observed on
result in excessive lacrimation, salivation, mucopurulent postmortem exam include necrotic and ulcerated mucous
discharge, and labored breathing (Fig. 91.2). Diarrhea is pos- membranes of the eyes, nose, and oropharynx (Fig. 91.3),
sible when gastrointestinal mucosae are affected. Secondary and tracheal congestion with blood-tinged foam. The lungs
644 SE C T I O N 18 Ruminants
share a common major antigen for neutralizing antibodies, passages in cell culture, are favored over killed vaccines, as
and it is thus not possible to distinguish SPPV, GTPV, or the latter do not provide adequate and long-lasting immu-
LSDV antibodies from one another.4 Immunity to CaPV nity.25 Several locally produced SPPV and GTPV vaccines
infection is mainly cell mediated, and infected animals are available using virus strains from Russia, Yugoslavia,
may produce only low levels of neutralizing antibodies. A Romania, and countries in Africa and the Indian subconti-
virus neutralization test is insufficiently sensitive to detect nent.5,21 In general, SPPV and GTPV are considered very
these levels.8 Other serologic tests show cross-reaction with host-specific, but a number of strains have been known
other poxviruses or are simply too difficult and expensive to to infect both sheep and goats. Some naturally occurring
perform.22,23 Several enzyme-linked immunosorbent assay recombinant strains are used for vaccines to protect both
(ELISA) tests have been reported, but currently no validated sheep and goats.26
ELISA for detecting antibodies against CaPV is available.4,8 There are currently three LSDV vaccines produced in
South Africa. Two of the vaccines contain strains of the
Treatment, Prevention, and Control original LSDV Neethling strain. The third vaccine uses
an attenuated South African LSDV field isolate.5 Due to
Other than supportive care, there is no specific treatment antigenic homology and cross-protection between CaPV,
available for CaPV infection. The clinical management SPPV and GTPV vaccines are used against LSDV only
of infected animals includes symptomatic treatment and in countries where the three CaPV overlap. A dose higher
strong antibiotic therapy to limit and control secondary than originally indicated is commonly used, but incomplete
bacterial infections. Potentially susceptible animals showing vaccine protection has been reported.5,26 In case of an emer-
typical signs of infection should be separated from others gency scenario in a CaPV-free country, killed vaccines are
or isolated if possible. Only prophylactic vaccination might recommended and safe to use.5 Currently, no Differentiat-
protect susceptible wild hoofstock. ing Infected from Vaccinated Animals (DIVA) vaccines are
Due to poor treatment options, prevention of virus commercially available against CaPV. Therefore, these need
introduction into hoofstock herds is crucial. Capripox-free to be developed for use in nonendemic countries.5
countries may maintain their disease-free status by respecting To date, there are no data published on the use of
strict import restrictions on livestock and animal products CaPV vaccines in nondomestic ruminants. The empiric
from affected areas. In countries where viruses are enzootic, use of Kenyavac, a live attenuated GTPV and SPPV vaccine
sanitary prophylactic measurements should be respected to containing the KSGP 0240 strain, in wild ruminants by the
avoid infection. Due to the highly infectious character of author proved effective in Laristan mouflon (Ovis orientalis
SPPV and GTPV, contact between exotic ruminants and laristanica). Nonetheless, its efficiency in other species is
domestic sheep and goats should be limited, and a strict unclear (unpublished data). Others in the Arabian Gulf
quarantine policy should be respected. Arthropod vector region have used vaccines in wild ruminants with vari-
control measures may minimize the risk of LSDV infection. able success (T. Cavero, personal communication, January
In case of an outbreak, successful control relies on early 16, 2017).
detection, a strict stamping-out and movement control
policy, and quarantine.5 However, the culling of rare
species valuable for conservation might be controversial, References
and this measure needs to be evaluated on a case-by-case
basis. Proper disposal and destruction of dead animals is 1. Buller RM, Arif BM, Black DN, et al: Family Poxviridae. In
Fauquet CM, Mayo MA, Maniloff J, et al, editors: Virus tax-
crucial, as is thorough cleaning and disinfection of the
onomy: Classification and nomenclature of viruses. Eighth report
premises and materials. The viruses are sensitive to phenol of the international committee on taxonomy of viruses, San Diego,
(2%), ether (20%), chloroform, formalin (1%), Virkon 2%, 2005, Elsevier Academic Press, pp 117–133.
iodine compounds (1:33 dilution), sodium hypochlorite 2. Diallo A, Viljoen GJ: Genus Capripoxvirus. In Mercer AA,
(2%–3%), sodium dodecyl sulfate, and quaternary ammo- Schmidt A, Weber O, editors: Poxviruses, Basel, 2007, Birkhäuser,
nium compounds (0.5%).22,23 In addition, vector control pp 167–181.
management is recommended for the prevention of LSDV 3. Babiuk S, Bowden TR, Boyle DB, et al: Capripoxviruses: an
transmission. emerging worldwide threat to sheep, goats and cattle, Transbound
Vaccination is the most effective way to control the Emerg Dis 55(7):263–272, 2008.
spread of CaPV in domestic ruminants. However, very little 4. Rao TVS, Bandyopadhyay SK: A comprehensive review of goat
is known about the use and effectiveness of vaccines in pox and sheep pox and their diagnosis, Anim Health Res Rev
1(02):127–136, 2000.
wildlife or their immune response. There are currently no
5. Tuppurainen ESM, Venter EH, Shisler JL, et al: Review: Cap-
vaccines registered for animals other than sheep, goats, and ripoxvirus diseases: current status and opportunities for control,
cattle. Only live attenuated vaccines are available, and none Transbound Emerg Dis 2015. 10.1111/tbed.1244.
of these are authorized for use in nonendemic countries. 6. Pieters W, Le Grange F, Arif A, et al: An outbreak of goat pox
Live and inactivated SPPV and GTPV vaccines have been virus in nondomestic hoofstock at Al Wabra Wildlife Preser-
reported, using different strains or isolates of the viruses. vation. In Proceedings of the 1st Joint AAZV/EAZWV/IZW
Live vaccines, which have been attenuated by multiple Conference, 2016, p 31.
646 SE C T I O N 18 Ruminants
7. Regnery RL: Poxviruses and the passive quest for novel hosts, in the Kruger National Park and Hluhluwe-iMfolozi Park, South
Curr Top Microbiol Immunol 315:345–361, 2007. Africa, J S Afr Vet Assoc 85(1):1–7, 2014.
8. Tuppurainen ESM, Oura CAL: Review: lumpy skin disease: an 18. Bowden TR, Babiuk SL, Parkyn GR, et al: Capripoxvirus tissue
emerging threat to Europe, the Middle East and Asia, Transbound tropism and shedding: a quantitative study in experimentally
Emerg Dis 59(1):40–48, 2012. infected sheep and goats, Virol 371(2):380–393, 2008.
9. Ali AA, Esmat M, Attia H, et al: Clinical and pathological studies 19. Chihota CM, Rennie LF, Kitching RP, et al: Mechanical trans-
on lumpy skin disease in Egypt, Vet Rec 127(22):549–550, mission of lumpy skin disease virus by Aedes aegypti (Diptera:
1990. Culicidae), Epidemiol Infect 126(2):317–321, 2001.
10. El-Nahas EM, El-Habbaa AS, El-bagoury GF, et al: Isolation 20. Carn VM, Kitching RP: An investigation of possible routes of
and identification of lumpy skin disease virus from naturally transmission of lumpy skin disease virus (Neethling), Epidemiol
infected buffaloes at Kaluobia, Egypt, Glob Vet 7:234–237, Infect 114(1):219–226, 1995.
2011. 21. Carn VM: Control of capripoxvirus infection, Vaccine 11(13):
11. Le Goff C, Lamien CE, Fakhfakh E, et al: Capripoxvirus 1275–1279, 1993.
G-protein-coupled chemokine receptor: a host-range gene suit- 22. World Organisation for Animal Health (OIE): Lumpy skin
able for virus animal origin discrimination, J Gen Virol 90(8): disease. Technical disease card, Paris, 2013, OIE. Available at:
1967–1977, 2009. http://www.oie.int/animal-health-in-the-world/technical-disease
12. Greta A, Gourreau JM, Vassart M, et al: Capripoxvirus disease in -cards/. (Accessed 9 January 2017).
an Arabian oryx (Oryx leucoryx) from Saudi Arabia, J Wildl Dis 23. World Organisation for Animal Health (OIE): Sheep pox and goat
28(2):295–300, 1992. pox. Technical disease card, Paris, 2013, OIE. Available at: http://
13. Young E, Basson PA, Weiss KE: Experimental infection of www.oie.int/animal-health-in-the-world/technical-disease-
game animals with lumpy skin disease virus (prototype strain cards/. (Accessed 9 January 2017).
Neethling), Onderstepoort J Vet Res 37:79–87, 1970. 24. World Organisation for Animal Health (OIE): Chapter 2.7.14.
14. Davies FG: Observations on the epidemiology of lumpy skin Sheep pox and Goat pox. In Manual of diagnostic test and vac-
disease in Kenya, J Hyg 88(1):95–102, 1982. cines for terrestrial animals, ed 6, Paris, 2016, OIE. Available at:
15. Hedger RS, Hamblin C: Neutralising antibodies to lumpy skin http://www.oie.int/en/international-standard-setting/terrestrial-
disease virus in African wildlife, Comp Immunol Microbiol Infect manual/access-online/. (Accessed 11 January 2017).
Dis 6(3):209–213, 1983. 25. Bhanuprakash V, Hosamani M, Venkatesan G, et al: Animal
16. Barnard BJH: Antibodies against some viruses of domestic poxvirus vaccines: a comprehensive overview, Expert Rev Vaccines
animals in southern African wild animals, Onderstepoort J Vet 11(11):1355–1374, 2012.
Res 64:95–110, 1997. 26. Tuppurainen ESM, Pearson CR, Bachanek-Bankowska K, et al:
17. Fagbo S, Coetzer JAW, Venter EH: Seroprevalence of Rift Valley Characterization of sheep pox virus vaccine for cattle against
fever and lumpy skin disease in African buffalo (Syncerus caffer) lumpy skin disease virus, Antiviral Res 109:1–6, 2014.
92
Babesiosis in Cervidae
ADRIANA PASTOR AND ELLIE MILNES
647
TABLE
92.1 Babesia spp. Identified Globally in Cervids
Clinical Signs
Babesia Geographic Wild or (Present/
Host Species spp.* Location Captive Animal Absent) Diagnostic Method Reference
European reindeer B. odocoilei Ontario, Canada Captive Present PCR and sequencing Pastor et al. (2016)27
(Rangifer tarandus
tarandus)
Quebec, Canada Captive Present PCR and sequencing Benoit et al. (2014)28
648 SE C T I O N 18 Ruminants
Manitoba, Canada Captive Present PCR and sequencing Mathieu et al. (2018)59
Pennsylvania, USA Captive Present PCR and sequencing Schoelkopf et al. (2005)7
New York, USA Captive Present PCR and sequencing Schoelkopf et al. (2005)7
Bartlett et al. (2009)6
Wisconsin, USA Captive Present PCR and sequencing Holman et al. (2003)10
B. odocoilei- Germany Captive Absent PCR and sequencing Wiegmann et al. (2015)13
like
B. divergens† Scotland, UK Captive Present PCR and sequencing Langton et al. (2003)25
Germany Captive Absent PCR and sequencing Wiegmann et al. (2015)13
B. venatorum Netherlands Captive Present PCR and sequencing Kik et al. (2011)3
Germany Captive Absent PCR and sequencing Wiegmann et al. (2015)13
Switzerland Captive Present PCR and sequencing Robert et al. (2008)12
B. capreoli Netherlands Captive Present PCR and sequencing Bos et al. (2016)2
Germany Captive Absent PCR and sequencing Wiegmann et al. (2015)13
B. divergens Germany Captive Absent PCR and sequencing Wiegmann et al. (2015)13
Babesia sp. California, USA Captive Absent In vitro culture Holman et al. (2002)29
PCR and sequencing Kjemtrup et al. (2000)30
IFA
B. jakimovi Russia Semi- Present Parasite morphology Nikol’skii et al. (1997)31
domesticated Holman et al. (2002)29
Woodland caribou B. odocoilei Minnesota, USA Captive Present Protozoal culture Holman (1994)32
(Rangifer tarandus PCR and sequencing Holman et al. (2000)4
caribou) Petrini et al. (1995)11
American elk B. odocoilei Ontario, Canada Captive Present PCR and sequencing Pastor et al. (2016)27
(Cervus elaphus
canadensis)
Saskatchewan, Captive Present PCR and sequencing Pattullo et al. (2013)5
Canada
New York, USA Captive Present PCR and sequencing Ameri et al. (2012)33
New Hampshire, USA Captive Present PCR and sequencing Schoelkopf et al. (2005)7
Quebec, Canada Captive Present PCR Benoit et al. (2014)28
Indiana, USA Captive Present Protozoal culture Gallatin et al. (2003)23
IFA
PCR and sequencing
Texas, USA Captive Present Protozoal culture Holman et al. (1994)34
PCR and sequencing Holman et al. (2000)4
Wisconsin, USA Captive Absent PCR and sequencing Holman et al. (2003)10
White-tailed deer B. odocoilei New Mexico, USA Wild Present (rare) Parasite morphology Spindler (1958)35
(Odocoileus
virginianus)
Texas, USA Wild Absent Parasite morphology Emerson and Wright (1968,
IFA 1970)36,37
PCR and sequencing Waldrup et al. (1992)38
Ramos et al. (2010)39
Tennessee, USA Wild Absent PCR and sequencing Fritzen et al. (2014)40
Oklahoma, USA Wild Absent IFA Waldrup et al. (1989)22
Virginia, USA Wild Absent Parasite morphology Perry et al. (1985)9
Minnesota, USA Captive Absent Protozoal culture Holman et al. (2000)4
IFA
PCR and sequencing
B. bigemina Texas, USA Wild Absent PCR and sequencing Holman et al. (2011)41
Mexico Wild Absent PCR and sequencing Cantu et al. (2007)42
B. cf. bovis Texas, USA Wild Absent PCR and sequencing Ramos et al. (2010)39
Mexico Wild Absent PCR and sequencing Cantu et al. (2007)42
Roe deer B. capreoli Italy Wild Absent PCR and sequencing Zanet et al. (2014)43
(Capreolus capreolus)
Netherlands Captive Present Parasite morphology Dorrestein et al. (1996)18
Germany Wild Absent PCR and sequencing Overzier et al. (2013)44
Poland Wild Absent PCR and sequencing Welc-Falęciak et al. (2013)45
Switzerland Wild Absent PCR and sequencing Hoby et al. (2009)15
Michel et al. (2014)46
France Wild Absent PCR and sequencing Bastian et al. (2012)47
B. venatorum France Wild Absent Protozoal culture Bonnet (2007)48
PCR and sequencing Bastian et al. (2012)47
Italy Wild Absent PCR and sequencing Zanet et al. (2014)43
Germany Wild Absent PCR and sequencing Overzier et al. (2013)44
Poland Wild Absent PCR and sequencing Welc-Falęciak et al. (2013)45
Switzerland Wild Absent PCR and sequencing Michel et al. (2014)46
Slovenia Wild Absent PCR and sequencing Duh (2005)49
B. divergens† Slovenia Wild Absent PCR and sequencing Duh (2005)49
Spain Wild Absent PCR and sequencing Garcia-Sanmartin et al.
(2007)50
B. bigemina Italy Wild Absent PCR and sequencing Zanet et al. (2014)43
Red deer (Cervus B. divergens† Slovenia Wild Absent PCR and sequencing Duh (2005)49
elaphus elaphus)
Ireland Wild Absent PCR and sequencing Zintl et al. (2011)51
Switzerland Wild Absent PCR and sequencing Michel et al. (2014)46
B. capreoli Italy Wild Absent PCR and sequencing Zanet et al. (2014)43
Switzerland Wild Absent PCR and sequencing Hoby et al. (2009)15
Scotland Wild NA Parasite morphology Gray et al. (1990)52
B. pecorum Spain Captive Absent PCR and sequencing Jouglin et al. (2014)53
B. bigemina Italy Wild Absent PCR and sequencing Zanet et al. (2014)43
B. divergens Austria Wild Absent PCR and sequencing Silaghi et al. (2011)54
CHAPTER 92 Babesiosis in Cervidae
Continued
649
TABLE
92.1 Babesia spp. Identified Globally in Cervids—cont’d
Clinical Signs
650 SE C T I O N 18 Ruminants
in Switzerland.2,14 Various Babesia spp. are found in Euro- infection may progress to clinical disease in the face of
pean red deer (Cervus elaphus elaphus), moose (Alces alces), concurrent stressors.9 High population density, comorbid
fallow deer (Dama dama), and also in roe deer (see Table disease, recent transport, reproductive status (rut, calving),
92.1).15–17 Clinical disease has not been reported in these and poor nutrition have been identified as potential risk
species, with the exception of a single report of clinical factors in the development of clinical disease.23
hemolytic disease in a captive roe deer in the Netherlands
attributed to B. capreoli based on parasite morphology; no
molecular diagnostics were performed in this case.18 In Pathology
northern Europe, sporadic cases of clinical babesiosis are Clinical Pathology
reported predominantly in captive reindeer and are associ-
ated with B. venatorum and B. capreoli infection. However, Romanowsky-type staining of a peripheral thin blood smear
a German polymerase chain reaction (PCR) survey found may not consistently identify intraerythrocytic life stages.
that 23.6% of clinically healthy zoo reindeer were hosts When present, Babesia appear as either single or paired
to five different Babesia spp., suggesting that subclinical piriform and ring-shaped organisms, often at the periphery
infections also occur in this species.13 of the erythrocyte (accolé position; Fig. 92.1) or in Maltese
cross arrangements. The number of parasites visible in blood
Factors Involved in Disease Emergence smears is extremely variable due to the removal of infected
erythrocytes: parasitemia of up to 80% of red blood cells
Climate change is an important driver of tick population may be observed in acute babesiosis, but parasitemia is not
dynamics and may aid the range expansion of vector-borne always visible on a blood smear. In addition, intraeryth-
pathogens into habitats that were previously too cold to rocytic Babesia are sometimes identified on routine blood
support the vectors.19 The large-scale seasonal movements smears of clinically normal animals.13
of migratory birds provide opportunities for bird-associated Hematology of animals with clinical babesiosis typically
ectoparasites and their pathogens to disperse rapidly over shows a normocytic, normochromic hemolytic anemia with
thousands of kilometers.19 This combination of factors, low red blood cell and hemoglobin values. An inflammatory
along with the presence of suitable local host species, has leukogram may also be present. Serum is often hemolyzed
resulted in the establishment of ticks and the diseases they or icteric, which may interfere with biochemistry results.
carry in new geographic regions (see also Chapter 36). No biochemistry changes are characteristic of Babesia
infection, but hyperbilirubinemia and bilirubinuria are
Zoonotic Potential often seen secondary to extravascular hemolysis, and in
severe disease azotemia occurs secondary to hemoglobinuric
Only one of the Babesia spp. reported to cause clinical nephropathy. Metabolic acidosis may result from increased
babesiosis in cervids, B. venatorum, is considered zoonotic. lactate generation secondary to tissue hypoxia.
At least three clinical cases of B. venatorum infection have
been reported in immunocompromised human patients in Gross Pathology
Europe.20 Clinical manifestation of the disease in humans
was a moderately severe malaria-like syndrome.21 Gross postmortem findings are not specific for babesiosis and
may include hepatomegaly; splenomegaly; hemoglobinuria
Clinical Signs
Sporadic cases, epizootics, and clinically silent infections
have all been described in captive cervids. Clinical presenta-
tion of cervid babesiosis has been described as consisting of
four syndromes: peracute, acute, chronic, and subclinical.
Peracute presentation is characterized by sudden death with
no prodromal signs. Acute cases may manifest with any
of the following: hemolysis, hemoglobinuria, hemorrhage,
icterus, lethargy, anorexia, or respiratory distress. At the
Toronto Zoo, separation of individuals from the herd
was noted as an early clinical sign in several reindeer and
one elk (Pastor, unpublished). Chronic cases have been
identified with any or all of the following clinical signs:
pyrexia, emaciation, anemia, and low-level parasitemia of
erythrocytes.22 Subclinical cases may experience transient
anemia, as reported in experimentally infected white-tailed • Figure 92.1 Babesia odocoilei infected erythrocytes on peripheral
deer, but notably, clinical signs do not always occur upon blood smear from a European reindeer (Rangifer tarandus). Arrow indi-
first exposure to the parasite; rather, subclinical or persistent cates organisms in the accolé position. (Courtesy Adriana R. Pastor.)
652 SE C T I O N 18 Ruminants
• Figure 92.2 Hemoglobinuria in the bladder of a European reindeer • Figure 92.4 Pigmentary nephrosis (diffuse dark red to black
(Rangifer tarandus) with clinical Babesia odocoilei infection. (Courtesy
discoloration of the kidneys) in the kidney of a European reindeer
Adriana R. Pastor.)
(Rangifer tarandus) with fatal Babesia odocoilei infection. (Courtesy
Adriana R. Pastor.)
Diagnosis
Antemortem diagnosis is based on the presentation of
typical clinical signs, hematology and biochemistry changes
consistent with hemolysis, the presence of intraerythrocytic
parasites on peripheral blood smear, or conventional PCR
on whole blood. However, intraerythrocytic Babesia may
not be identified on peripheral blood smear, even in clini-
cal cases, complicating diagnosis. Postmortem diagnosis is
• Figure 92.3 Endocardial hemorrhage in the heart of a European based on the presence of hemolysis, extensive hemorrhage,
reindeer (Rangifer tarandus) with clinical Babesia odocoilei infection. hemoglobinuria, and icterus. PCR of DNA extracted from
(Courtesy Adriana R. Pastor.) fresh-frozen spleen samples has been used to successfully
diagnose the disease in deceased elk, reindeer, and white-
(Fig. 92.2); icterus; petechial hemorrhages, particularly of tailed deer, including tissues that have been stored at –20°C
the endocardium (Fig. 92.3) and adrenal glands; and diffuse for up to 6 years (Pastor, unpublished). The use of PCR
dark red to black discoloration of the kidneys (pigmentary on routine blood samples or banked frozen spleen samples
nephrosis; Fig. 92.4). could potentially aid in disease surveillance efforts and
detection of subclinically infected wild and captive cervids.
Histopathology Recently, sequencing of the mitochondrial COI region has
been used to identify multiple strains of B. odocoilei in
Histologic tissue changes are variable and are generally affected cervids at the Toronto Zoo (Pastor, unpublished),
consistent with hemolytic disease. Impression smears of and similar techniques may prove useful in determining the
parenchymatous organs may reveal intraerythrocytic life epidemiology of the disease in affected areas.
stages. Hepatic centrilobular vacuolar degeneration and Prior to the development of molecular characterization
necrosis may occur due to decreased hepatic perfusion and of protozoal organisms, identification of piroplasms was
oxygenation secondary to hemolytic anemia. Splenic hemo- based primarily on morphologic and morphometric char-
siderosis and erythrophagocytosis and generalized lymph acteristics of the different protozoal life stages. Serology and
node erythrophagocytosis reflect extravascular hemolysis. indirect fluorescent antibody assay have also been used to
In the kidney, hemoglobinuric nephropathy (pigmentary identify Babesia spp. infection. Due to the great degree of
nephrosis with acute tubular degeneration) and dilated similarity between piroplasms in size and appearance, and
renal tubules containing granular hemoglobin casts may the variable specificity of antibody binding, it is likely that
CHAPTER 92 Babesiosis in Cervidae 653
some Babesia spp. historically reported in cervids were mis- permethrin at the recommended label dose for cattle have
identified. Molecular diagnostics have allowed for improved been used safely and effectively in zoo cervids in order to
identification of different Babesia spp.24 For example, a control ticks; ivermectin at 0.4 mg/kg BW (double the cattle
Babesia sp. isolated from an outbreak in a Scottish reindeer dose) may be administered subcutaneously or orally.23,26
herd, identified by the authors as B. divergens, is now The use of babesiacidal compounds, in combination with
thought more likely to have been caused by B. capreoli.24,25 acaricide treatment, is recommended when translocating
Submission of Babesia isolates from cervid babesiosis cases cervids to and from Babesia-endemic areas. Cervid transloca-
for molecular diagnostics is strongly encouraged, to advance tions should be designed to avoid widening the geographic
species identification and further elucidate the epidemiol- range of this parasite, and to protect naïve populations
ogy of the disease. from Babesia infection. Stress has been shown to precipitate
acute hemolytic disease in animals harboring subclinical
Treatment infections, so efforts should be made to reduce stress where
possible. Strategic treatment of subclinically or potentially
A number of chemical compounds have been reported infected animals with babesiacidal compounds may be a
to be effective against Babesia spp. in domestic animals, useful prophylactic measure when stress is anticipated.
including diminazene diaceturate, imidocarb dipropionate,
and amicarbalide.8 These drugs are not available world- References
wide, and attempts to obtain antiprotozoal drugs in an
1. Homer MJ, Aguilar-Delfin I, Telford SR, III, et al: Babesiosis,
emergency may be unsuccessful. Dose rates and frequency Clin Microbiol Rev 13(3):451, 2000.
of administration have not been established for most drugs 2. Bos JH, Klip FC, Sprong H, et al: Babesia capreoli infection in
when used in cervids; therefore therapeutic regimens are captive reindeer (Rangifer tarandus tarandus) in Ouwehand Zoo,
based on the manufacturer’s recommendations for cattle. the Netherlands. American Association of Zoo Veterinarians
Imidocarb dipropionate at 2.2–3.0 mg/kg body weight Annual Conference, Atlanta, GA, 2016.
(BW) by intramuscular injection has been used successfully 3. Kik M, Nijhof AM, Balk JA, et al: Babesia sp. EU1 infection in
in captive elk and reindeer to treat clinical disease and a forest reindeer, The Netherlands, 2011.
eliminate subclinical Babesia infections.6,25 Following treat- 4. Holman PJ, Madeley J, Craig TM, et al: Antigenic, phenotypic
ment with babesiacidal compounds, anemia may worsen for and molecular characterization confirms Babesia odocoilei isolated
up to 7 days due to the continued removal of parasitized from three cervids, J Wildl Dis 36(3):518, 2000.
5. Pattullo KM, Wobeser G, Lockerbie BP, et al: Babesia odocoilei
erythrocytes from circulation.6
infection in a Saskatchewan elk (Cervus elaphus canadensis) herd,
Successful treatment depends on early diagnosis and J Vet Diagn Invest 25(4):535–540, 2013.
prompt administration of babesiacides. The prognosis is 6. Bartlett SL, Abou-Madi N, Messick JB, et al: Diagnosis and
grave in animals that are debilitated by acute disease.8 treatment of Babesia odocoilei in captive reindeer (Rangifer
Aggressive supportive treatment is required in animals tarandus tarandus) and recognition of three novel host species, J
undergoing hemolytic crisis, and should include anti- Zoo Wildl Med 40(1):152–159, 2009.
inflammatory drugs and fluid therapy to minimize the 7. Schoelkopf L, Hutchinson CE, Bendele KG, et al: New ruminant
secondary renal effects of hemolysis. Based on clinical hosts and wider geographic range identified for Babesia odocoilei
experience with domestic ungulates, blood transfusions (Emerson and Wright 1970), J Wildl Dis 41(4):683–690, 2005.
may be life-saving in very anemic animals, although this 8. Vial HJ, Gorenflot A: Chemotherapy against babesiosis, Vet
technique has not yet been reported as part of the clinical Parasitol 138:147–160, 2006.
9. Perry BD, Nichols DK, Cullom ES: Babesia odocoilei (Emerson
management of babesiosis in cervids. Donor animals will
and Wright 1970) in white-tailed deer, Odocoileus virginianus
ideally be well-conditioned, and the packed cell volume of (Zimmermann), in Virginia, J Wildl Dis 21(2):149, 1985.
the donor should be checked before collecting blood. No 10. Holman P, Bendele K, Schoelkopf L, et al: Ribosomal RNA
published information on known blood groups in cervids analysis of Babesia odocoilei isolates from farmed reindeer (Ran-
could be found at the time of writing, but use of related gifer tarandus tarandus) and elk (Cervus elaphus canadensis) in
donor animals may be advisable. Wisconsin, Parasitol Res 91(5):378–383, 2003.
11. Petrini KR, Holman PJ, Rhyan JC, et al: Fatal babesiosis in an
Prevention and Control American woodland caribou (Rangifer tarandus caribou), J Zoo
Wildl Med 26(2):298–305, 1995.
Complete eradication of the tick vector from the environ- 12. Robert N, Hoby S, Mathis A, et al: Fatal babesiosis caused by
ment is rarely feasible, and consideration should be given Babesia sp. EU1 in a reindeer at the Zoo Basel, Switzerland.
European Association of Zoo and Wildlife Veterinarians 7th
to maintaining enzootic stability in endemic regions. In a
Scientific Meeting, Leipzig, Germany, 2008.
captive setting, tick control programs should be designed to 13. Wiegmann L, Silaghi C, Obiegala A, et al: Occurrence of Babesia
reduce the risk of tick populations developing resistance to species in captive reindeer (Rangifer tarandus) in Germany, Vet
acaricides. Wild deer should be excluded from outdoor zoo Parasitol 211(1):16–22, 2015.
exhibits. A strategic tick control program should integrate 14. Hoby S, Robert N, Mathis A, et al: Babesiosis in free-ranging
pasture management (removal of bushes and long grass) chamois (Rupicapra r. rupicapra) from Switzerland, Vet Parasitol
with application of acaricides. Both topical amitraz and 148(3):341–345, 2007.
654 SE C T I O N 18 Ruminants
15. Hoby S, Mathis A, Doherr MG, et al: Babesia capreoli infections 34. Holman PJ, Craig TM, Crider DLD, et al: Culture isolation and
in alpine chamois (Rupicapra r. rupicapra), roe deer (Capreolus c. partial characterization of a Babesia sp. from a North American
capreolus) and red deer (Cervus elaphus) from Switzerland, J Wildl elk (Cervus elaphus), J Wildl Dis 30(3):460–465, 1994.
Dis 45(3):748–753, 2009. 35. Spindler LA, Allen RW, Diamond LS, et al: Babesia in a white-
16. Pūraitė I, Rosef O, Radzijevskaja J, et al: The first detection of tailed deer, J Protozool 5:8, 1958.
species of Babesia Starcovici, 1893 in moose, Alces alces (Lin- 36. Emerson HR, Wright WT: The isolation of a Babesia in white-
naeus), in Norway, Folia Parasitol (Praha) 63:009, 2016. tailed deer, Wildl Dis 4(4):142–143, 1968.
17. Rehbein S, Visser M, Jekel I, et al: Endoparasites of the fallow 37. Emerson HR, Wright WT: Correction, J Wildl Dis 6:519,
deer (Dama dama) of the Antheringer Au in Salzburg, Austria, 1970.
Wien Klin Wochenschr 126(Suppl 1):37–41, 2014. 38. Waldrup KA, Moritz J, Baggett D, et al: Monthly incidence
18. Dorrestein GM, Jongejan F, Rijpkema S: Survey of tick related of Theileria cervi and seroconversion to Babesia odocoilei in
problems in roe deer (Capreolus capreolus) in The Netherlands, white-tailed deer (Odocoileus virginianus) in Texas, J Wildl Dis
Vet Q 18(Suppl 3):148, 1996. 28(3):457–459, 1992.
19. Ogden NH, Maarouf A, Barker IK, et al: Climate change and the 39. Ramos CM, Cooper SM, Holman PJ: Molecular and serologic
potential for range expansion of the Lyme disease vector Ixodes evidence for Babesia bovis-like parasites in white-tailed deer
scapularis in Canada, Int J Parasitol 36(1):63–70, 2006. (Odocoileus virginianus) in south Texas, Vet Parasitol 172(3):
20. Herwaldt BL, Cacciò S, Gherlinzoni F, et al: Molecular 214–220, 2010.
characterization of a non-Babesia divergens organism causing 40. Fritzen C, Mosites E, Applegate RD, et al: Environmental inves-
zoonotic babesiosis in Europe, Emerg Infect Dis 9(8):942–948, tigation following the first human case of babesiosis in Tennessee,
2003. J Parasitol 100(1):106–109, 2014.
21. Gray J, Zintl A, Hildebrandt A, et al: Zoonotic babesiosis: 41. Holman PJ, Carroll JE, Pugh R, et al: Molecular detection of
overview of the disease and novel aspects of pathogen identity, Babesia bovis and Babesia bigemina in white-tailed deer (Odocoi-
Ticks Tick Borne Dis 1(1):3–10, 2010. leus virginianus) from Tom Green County in central Texas, Vet
22. Waldrup KA, Kocan AA, Qureshi T, et al: Serological preva- Parasitol 177(3):298–304, 2011.
lence and isolation of Babesia odocoilei among white-tailed deer 42. Cantu A, Ortega-S JA, Henke SE, et al: Immunologic and
(Odocoileus virginianus) in Texas and Oklahoma, J Wildl Dis molecular identification of Babesia bovis and Babesia bigemina
25(2):194, 1989. in free-ranging white-tailed deer in Northern Mexico, J Wildl
23. Gallatin LL, Sojka JE, Irizarry-Rovira AR, et al: Babesia odocoilei Dis 43(3):504–507, 2007.
infection in elk, J Am Vet Med Assoc 223(7):1027–1986, 2003. 43. Zanet S, Trisciuoglio A, Bottero E, et al: Piroplasmosis in wild-
24. Malandrin L, Jouglin M, Sun Y, et al: Redescription of Babesia life: Babesia and Theileria affecting free-ranging ungulates and
capreoli (Enigk and Friedhoff, 1962) from roe deer (Capreolus carnivores in the Italian Alps, Parasit Vectors 7(1):70, 2014.
capreolus): isolation, cultivation, host specificity, molecular 44. Overzier E, Pfister K, Herb I, et al: Detection of tick-borne
characterisation and differentiation from Babesia divergens, Int J pathogens in roe deer (Capreolus capreolus), in questing ticks
Parasitol 40(3):277–284, 2010. (Ixodes ricinus), and in ticks infesting roe deer in southern
25. Langton C, Gray J, Waters P, et al: Naturally acquired babesiosis Germany, Ticks Tick Borne Dis 4(4):320–328, 2013.
in a reindeer (Rangifer tarandus tarandus) herd in Great Britain, 45. Welc-Falęciak R, Werszko J, Cydzik K, et al: Co-infection and
Parasitol Res 89(3):194–198, 2003. genetic diversity of tick-borne pathogens in roe deer from Poland,
26. Masters NJ, Flach E: Tragulidae, Moschidae, and Cervidae. In Vector Borne Zoonotic Dis 13(5):277–288, 2013.
Zoo and wild animal medicine, ed 8, St. Louis, MO, 2015, 46. Michel AO, Ryser-Degiorgis M-P, Mathis A: Babesia spp. in
Saunders, pp 611–625. European wild ruminant species: Parasite diversity and risk
27. Pastor AR, Nielsen AMW, Kirkbright DW, et al: Babesiosis factors for infection, Vet Res 45(1), 2014.
(Babesia odocoilei): An emerging disease of Ontario cervids. 47. Bastian S, Jouglin M, Brisseau N, et al: Antibody prevalence and
American Association of Zoo Veterinarians Annual Conference, molecular identification of Babesia spp. In roe deer in France, J
Atlanta, GA, 2016. Wildl Dis 48(2):416–424, 2012.
28. Benoit J-M, Proulx C, McKenzie D, et al: Premiers cas de babési- 48. Bonnet S: Babesia sp. EU1 from roe deer and transmission within
ose chez des cervidés du Quebec, Reseau d’Alerte et d’Information Ixodes ricinus, Emerging Infect Dis J 13(8), 2007.
Zoosanitaire Bulletin Zoosanitaire 1–5, 2014. 49. Duh D: Cervids as Babesiae hosts, Slovenia, Emerging Infect Dis
29. Holman P, Swift P, Frey R, et al: Genotypically unique Babesia J 11(7), 2005.
spp. isolated from reindeer (Rangifer tarandus tarandus) in the 50. García-Sanmartín J, Aurtenetxe O, Barral M, et al: Molecular
United States, Parasitol Res 88(5):405–411, 2002. detection and characterization of piroplasms infecting cervids
30. Kjemtrup A, Thomford J, Robinson T, et al: Phylogenetic rela- and chamois in Northern Spain, Parasitology 134(Pt 3):391,
tionships of human and wildlife piroplasm isolates in the western 2007.
United States inferred from the 18S nuclear small subunit RNA 51. Zintl A, Finnerty EJ, Murphy TM, et al: Babesias of red deer
gene, Parasitology 120:487–493, 2000. (Cervus elaphus) in Ireland, Vet Res 42(1):7, 2011.
31. Nikol’skii SN, Nikiforenko VI, Pozov SA: Epidemiology of 52. Gray JS, Murphy TM, Taylor SM, et al: Comparative mor-
piroplasmosis in Siberia, Veterinariya 4, 1977. phological and cross transmission studies with bovine and deer
32. Holman P: Comparative study of cervid Babesia isolates. In babesias in Ireland, Prev Vet Med 9(3):185–193, 1990.
Wagner GG, editors: ProQuest Dissertations Publishing; 53. Jouglin M, Fernández-de-Mera IG, de La Cotte N, et al: Isolation
1994. and characterization of Babesia pecorum sp. nov. from farmed red
33. Ameri M, Anderson W, Holman P, et al: Babesia odocoilei infec- deer (Cervus elaphus), Vet Res 45:78, 2014.
tion in a North American elk (Cervus elaphus canadensis), Comp 54. Silaghi C, Hamel D, Pfister K, et al: Babesia species and co-
Clin Pathol 21(3):363–365, 2012. infection with Anaplasma phagocytophilum in free-ranging
CHAPTER 92 Babesiosis in Cervidae 655
ungulates from Tyrol (Austria), Tierärztliche Mschr Vet Med and marsh deer (Blastocerus dichotomus) in the State of Minas
Austria 98:268–274, 2011. Gerais, Brazil, Vet Parasitol 177(1–2):61–66, 2011.
55. García-Vásquez Z, Ortega-S JA, Cantu-Covarruvias A, et al: 58. Thomford JW, Conrad PA, Boyce WM, et al: Isolation and
Tick-borne diseases in syntopic populations of fallow deer (Dama in vitro culture of Babesia parasites from free-ranging desert
dama) and axis deer (Axis axis) in northern Mexico, J Wildl Dis bighorn sheep (Ovis canadensis nelsoni) and mule deer (Odocoileus
51(2):527–529, 2015. hemionus) in California, J Parasitol 79(1):77–84, 1993.
56. Silveira JAG, Rabelo EML, Lacerda ACR, et al: Molecular 59. Mathieu A, Pastor A, Berkvens C, et al: Babesia odocoilei as
detection and identification of hemoparasites in pampas deer a cause of mortality in captive cervids in Canada, Can Vet J
(Ozotoceros bezoarticus Linnaeus, 1758) from the Pantanal Brazil, 59(1):52–58, 2018.
Ticks Tick Borne Dis 4(4):341, 2013.
57. Da Silveira JAG, Rabelo ÉML, Ribeiro MFB: Detection of Thei-
leria and Babesia in brown brocket deer (Mazama gouazoubira)
SECTION 19
656
93
Management of Dental Disease
in Elephants
GERHARD STEENKAMP
657
658 SE C T I O N 19 Elephants and Rhinoceroses
• Figure 93.3 The conically shaped pulp and the tusk it originated
from. This is a healthy tusk from an adult African elephant (Loxodonta
africana) bull. There is a clear distinction between the white ivory (that
was in the alveolus) and the brownish ivory of the exposed tusk.
• Figure 93.6 A long-standing complicated crown fracture (pulp • Figure 93.7 A rostro-caudal oblique radiograph of a 4-year-old
exposed) in a 17-year-old African elephant (Loxodonta africana) bull. Asian elephant (Elephas maximus) bull. In these smaller individuals,
radiography may assist in making a definitive diagnosis. Radiograph
reproduced with the kind permission of the Taronga Conservation
Society, Australia.
• Figure 93.8 Endoscopic evaluation of the pulp cavity (pulposcopy) • Figure 93.9 The left maxillary molar 4 in this adult Asian elephant
is another imaging technique that is valuable to help the clinician make (Elephas maximus) cow has rotated through 90 degrees and is now
a diagnosis. In this pulp cavity, only approximately 10 mm of pulp blocking the eruption of the molar behind it. Molar extraction is indi-
(thickness) was left after chronic pulpitis. This individual responded cated in these cases. (Photo reproduced with the kind permission of
poorly to a partial pulpectomy, and the tusk was extracted 1 month Dr. Chris Visser, Phoenix, AZ.)
later.
teeth or tooth fragments are often loose. After extraction of Diseases/Anomalies Affecting the Tusks
the molar fragment, the opposing molar should be evalu- Absent Tusks
ated and trimmed if necessary.
Elephants without visible tusks, especially individuals with
Supernumerary Molars unknown histories, should be evaluated for tusks or tusk
fragments. The causes of absent tusks may be due to lack
Supernumerary or seventh molars have been described from of tusk development (e.g., in Asian elephant cows), odonto-
mandibles of African elephants.25 In one skull I observed, genic tumor formation, or premature loss of the tusk. I am
the seventh molar was present in the mandible but had not aware of two different reports, one in Addo Elephant Park
erupted. (Dr. Markus Hofmeyr, personal communication, 2012) and
the other in the Johannesburg Zoo (Mr. Phillip Cronje,
Rotation personal communication, 2006), where individuals lost a
complete tusk. The cause of the tusk loss in the case of
Due to delayed shedding in Asian elephants, the piece of Addo bull is unclear, while the zoo elephant fell into the
molar that is still present in the mouth may rotate by up enclosure moat and suffered a complete avulsion of one of
to 90° and cause retention of the molar fragment and delay its tusks.
the eruption of the next molar in sequence (Fig. 93.9).23,24
These impacted molars may present with similar clinical Supernumerary Tusks
signs, as mentioned in delayed shedding, in addition to
impeding the eruption of the caudal molar. This condition was first described by Sir Frank Colyer in
the previous century.26,27 To my knowledge, no clinical case
Perforations/Cavities has ever been published.
Tusk fracture/crack
Explore
process to determine the correct treatment protocol to be used. This diagram represents the author’s
decision-making process when assessing fractured or cracked tusks.
• Figure 93.11 This 7-year-old African elephant (Loxodonta africana) bull fractured his left tusk at the
level of the skinfold after trying to open a steel gate. The author performed a partial pulpectomy within 7
days of the accident, and this picture, taken 2 years later, shows healthy tusk growth on the treated tusk.
664 SE C T I O N 19 Elephants and Rhinoceroses
References 15. Fagan DA, Benirschke K, Simon JH, et al: Elephant dental pulp
tissue: where are the nerves?, J Vet Dent 16:169–172, 1999.
1. Roca AL, Ishida Y, Brandt AL, et al: Elephant natural history: a 16. Weissengruber GE, Egerbacher M, Forstenpointner G: Structure
genomic perspective, Annu Rev Anim Biosci 3:139–167, 2015. and innervation of the tusk pulp in the African elephant (Lox-
2. Cerling TE, Wittemyer G, Ehleringer JR, et al: History of odonta africana), J Anat 206:387–393, 2005.
animals using isotope records (HAIR): a 6-year dietary history 17. Stansfield FJ: A novel objective method of estimating the age of
of one family of African elephants, Proc Natl Acad Sci USA mandibles from African elephants (Loxodonta africana africana),
106:8093–8100, 2009. PLoS ONE 10:e0124980, 2015.
3. Codron J, Codron D, Lee-Thorp JA, et al: Landscape-scale 18. Tsung AH, Allen BR: A 51-year-old woman crushed by an
feeding patterns of African elephant inferred from carbon isotope elephant trunk, Wilderness Environ Med 26:54–58, 2015.
analysis of feces, Oecologia 165:89–99, 2011. 19. Young AM, Joseph AP, Jackson A: Crush injury by an elephant:
4. Codron J, Codron D, Sponheimer M, et al: Stable isotope series life-saving prehospital care resulting in a good recovery, Med J
from elephant ivory reveal lifetime histories of a true dietary Aust 203:264–265 e261, 2015.
generalist, Proc Biol Sci 279:2433–2441, 2012. 20. Legendre LFJ: Vital pulpotomy of the broken tusk of an Indian
5. Greco BJ, Meehan CL, Miller LJ, et al: Elephant management elephant, J Vet Dent 10(10):11, 1993.
in North American zoos: environmental enrichment, feeding, 21. Robinson PT, Schmidt M: Dentistry in zoo animals: dental
exercise, and training, PLoS ONE 11:e0152490, 2016. diseases of elephants and hippos. In Fowler ME, editor: Zoo and
6. Steenkamp G, Ferreira SM, Bester MN: Tusklessness and tusk wild animal medicine, ed 2, Philadelphia, PA, 1986, Saunders,
fractures in free-ranging African savanna elephants (Loxodonta pp 534–547.
africana), J S Afr Vet Assoc 78:75–80, 2007. 22. Steenkamp G, Ferguson WH, Boy SC, et al: Estimating exposed
7. Raubenheimer EJ, van Heerden WF, van Niekerk PJ, et al: pulp lengths of tusks in the African elephant (Loxodonta africana
Morphology of the deciduous tusk (tush) of the African elephant africana), J S Afr Vet Assoc 79:25–30, 2008.
(Loxodonta africana), Arch Oral Biol 40:571–576, 1995. 23. Kertesz P: Dental diseases and their treatment in captive wild
8. Raubenheimer EJ: Early development of the tush and the tusk animals. In Kertesz P, editor: A colour atlas of veterinary dentistry
of the African elephant (Loxodonta africana), Arch Oral Biol and oral surgery, London, 1993, Wolfe Publishing, pp 215–281.
45:983–986, 2000. 24. Fagan DA, Oosterhuis JE, Roocroft A: Captivity disorders in
9. Kertesz P: Comparative odontology. In Kertesz P, editor: A colour elephants: impacted molars and broken tusks, Der Zoologische
atlas of veterinary dentistry and oral surgery, London, 1993, Wolfe Garten 71:281–303, 2001.
Publishing, pp 35–50. 25. Steenkamp G: Oral biology and disorders of tusked mammals,
10. Laws RM: Age criteria for the African elephant Loxodonta a. Vet Clin North Am Exot Anim Pract 6:689–725, 2003.
africana, Afr J Ecol 4:1–37, 1966. 26. Miles AEW, Grigson C: The ungulates. In Miles AEW, Grigson
11. Lee PC, Sayialel S, Lindsay WK, et al: African elephant age C, editors: Colyer’s variations and diseases of the teeth of animals,
determination from teeth: validation from known individuals, rev ed, Cambridge, 1990, Cambridge University Press, pp
Afr J Ecol 50:9–20, 2012. 106–129.
12. Scmitt DL: Proboscidea (Elephants). In Fowler M, Miller RE, 27. Sikes SK: The African elephant and its health. In Sikes SK, editor:
editors: Zoo and wild animal medicine, ed 5, 2003, Saunders, pp The natural history of the elephant, New York, 1971, American
541–550. Elsevier Company, pp 185–223.
13. Phuangkum P, Lair RC, Angkawanith T: Elephant care manual 28. Dorland WAN: Dorland’s illustrated medical dictionary. In
for mahouts and camp managers. In: Nations FaAOotU, Forest Dorland WAN, editor: Dorland’s illustrated medical dictionary,
Industry Organization MONRAE, eds. Bangkok: FAO & FIO, ed 32, Philadelphia, PA, 2012, Saunders/Elsevier, p 519.
2005. 29. Mutinda M, Chenge G, Gakuya F, et al: Detusking fence-breaker
14. Boy SC, Steenkamp G: Neural innervation of the tusk pulp of elephants as an approach in human-elephant conflict mitigation,
the African elephant (Loxodonta africana), Vet Rec 154:372–374, PLoS ONE 9:e91749, 2014.
2004.
94
Elephant Mycobacteriosis: New
Diagnostics and Management
KAY A. BACKUES AND ELLEN WIEDNER
I
nfection with Mycobacterium tuberculosis (Mtb) is now Aerosolized material from infected animals appears to be the
widely recognized as an important primarily respiratory most common source of transmission. Fomites and manure,
tract disease of Asian elephants, Elephas maximus, in significant sources for the transmission of M. bovis, do not
human care, but is rarely reported in captive African ele- appear to be of much importance for the transmission of
phants, Loxodonta africana.1 Mycobacterium bovis also occurs Mtb.
in elephants, but this chapter focuses on M. tuberculosis.
Between 1997 and 2011, the point prevalence of Mtb Diagnosis
in North America was 5.1% in Asian elephants but zero in
African elephants.1 The close working partnership between Screening for Mtb should be included in all preventative
humans and Asian elephants likely explains many of the medicine programs for captive elephants. This entails moni-
Asian elephant exposures to this human disease. However, toring individual elephants as well as overall herd health.
there have been two reported cases of confirmed Mtb in Keeper health, zoo-wide medical concerns, and general
free-ranging Asian elephants and one unconfirmed case in husbandry procedures also need to be considered.5 Diag-
an African elephant.2–4 Needless to say, the introduction nostic screening tests routinely used in humans and other
of Mtb into free-ranging populations of Asian elephants species—such as thoracic radiographs and lung and gastric
could pose yet another significant threat to this highly washes—are precluded in elephants by their great size and
endangered species. Extensive knowledge about Mtb’s anatomy and poorly studied immune systems. Intradermal
behavior, diagnosis, and treatment in its primary reservoir, testing with tuberculin causes nonspecific reactions and is
humans, has been used as a loose model for the disease in contraindicated in elephants.10 Multiple diagnostic tests are
Asian elephants.5 Such assumptions may provide a starting often necessary to screen for Mtb in living elephants, and
point, but they require the caveat that our understanding definitive antemortem confirmation of disease presence or
of Mtb in elephants is still in its infancy, and many of these absence may be elusive.
assumptions may not be correct. Direct tests such as culture and real-time polymerase
chain reaction (qPCR) confirm the actual presence of Mtb
Transmission complex organisms. Culture is the gold standard test for
Mtb diagnosis in all species and the only test that confirms
As occurs with transmission of Mtb between humans, active infection. For living elephants, a trunk wash (TW)
transmission of Mtb between elephants and between is commonly submitted for Mtb culture because it con-
elephants and humans requires close, prolonged aerosol tains material from the lower respiratory tract. A properly
contact with another infected individual.6–9 At one facil- performed TW necessitates training the elephant to permit
ity individuals that only had brief casual contact with an the instillation of 60 mL of saline into its trunk and then
infected elephant did not respond to tuberculin intradermal exhaling into a sterile collection container.11
testing.9 At another facility, numerous human intradermal Indirect tests such as serology, interferon gamma (IFN-γ)
test conversions occurred following use of a high-pressure tests, and cytokine stimulation assays demonstrate expo-
washer near a poorly placed intake vent for an office inside sure to Mtb complex organisms.12 Serologic tests identify
an elephant barn. The barn housed an Mtb-infected and antibodies to specific mycobacterial antigens, whereas
shedding elephant.7 The strong correlation between close, cytokine stimulation assays and IFN-γ tests detect specific
prolonged aerosol contact and disease transmission high- components of cell-mediated immunity (CMI) triggered
lights the necessity of stopping active shedding by infected by Mtb.13–16 However, indirect tests cannot confirm active
elephants as soon as possible via antibiotic treatment. infection. Most of the CMI tests are experimental, minimally
665
666 SE C T I O N 19 Elephants and Rhinoceroses
validated, and not available commercially.12 See Table 94.1 killing of actively replicating Mtb organisms within a few
for details of Mtb diagnostic tests. days of starting treatment and has been documented to
If an elephant is positive on an indirect test, confirmatory stop trunk shedding if compliance is good, the isolate is
direct tests are recommended, such as increased frequency INH-susceptible, and the dosing is sufficient.19 INH should
of TW cultures and qPCR (if available). If a positive TW not, however, be used as monotherapy because of the risk of
culture is obtained, confirm results either by submitting a developing Mtb resistance.20 The importance of RIF stems
duplicate retained specimen or by submitting new TW for from its ability to resolve cavitary lesions and activity against
cultures. Multiple TWs performed over weeks to months latent Mtb organisms. PZA and ETH are synergistic with
may be necessary to confirm a positive result due to low the other drugs and important in preventing resistance, but
bacterial numbers or intermittent shedding. Herdmates neither should be used in place of INH or RIF. LEVO can
should also undergo increased surveillance, and regulatory be substituted for PZA, ETH, or RIF but not for INH.
agencies should be contacted.5 There is evidence from human treatment regimens that
Positive cultures should be tested for their antibiotic sus- fluoroquinolones may be useful in situations involving resis-
ceptibility and the isolate sequenced for molecular typing. tance to first-line antibiotics or nonreplicating (latent) Mtb
Complete DNA sequencing of isolates can be performed organisms.21–23 Fluoroquinolones can be delivered orally or
upon request and can help to elucidate the epidemiology rectally, unlike RIF, which requires oral administration. See
of the infection. Molecular probes can identify genes associ- Table 94.2 for drug dosing information.
ated with drug resistance in elephant Mtb samples. If the The appropriate drug doses and concentrations needed
animal is treated but then relapses, these tests should be for cure in elephants are unknown; only the dosages needed
repeated to see if the susceptibility pattern has changed or to achieve specified blood concentrations.24–28 Individual
if a different Mtb isolate is involved.17 elephants may show significant variation in drug pharma-
As with many other aspects of elephant health, screen- cokinetics.28,29 Although the plasma/serum drug concentra-
ing over significant periods of time appears essential to truly tions used in humans are starting points, they may not
understanding a given herd’s Mtb status as opposed to using always be satisfactory for elephants and may even result in
a single time point’s test results. All elephants that die in a toxicity. Documented adverse events include depression,
collection should have a complete necropsy performed to colic, inappetance, and black fetid manure.30,31 Blepharo-
confirm their Mtb infection status. All necropsies should be spasm, ocular tearing, and lethargy are also reported. Never-
undertaken as if an elephant may be positive for Mtb, and for theless, the highest doses of antitubercular drugs should be
elephants known to have confirmed Mtb infection at death, given that do not cause toxicity in order to maximize drug
a risk-benefit analysis of human safety should be evaluated exposure at the infection site and to minimize development
before necropsy. In addition to infrequent shedding and of resistance.32–34
equivocal test results, long periods of latency appear likely Culture-positive elephants may be treated with a short
in Mtb-infected elephants, further challenging diagnosis. intensive phase (or initiation phase) and a longer continu-
ation phase. During the intensive phase, high and frequent
Treatment doses of at least three to four drugs are given to cause a rapid
decrease in the number of infectious organisms and to avoid
In treating an elephant for Mtb infection, several issues development of resistance. This phase may last 8–10 weeks
must be considered. The first is prevention of shedding of if tolerated by the elephant. Efficacy may be monitored via
infectious organisms into the environment, which is best frequent TW culture to document cessation of mycobacte-
accomplished using a combination of first-line antituber- rial shedding. In the continuation phase, antibiotics may
cular medications with consistent dosing at sufficient levels. be used at lower frequency but not necessarily lower doses
The second is avoidance of serious drug-related adverse over many months to kill remaining viable organisms.
events. The third is achieving and maintaining adequate Other treatment regimens exist, and several facilities have
serum drugs levels throughout treatment. used the same dose and frequency throughout an elephant’s
Treatment should be started as soon as an Mtb-infected treatment regimen. Treatment regimens vary, but all should
elephant is identified by culture, even before antibiotic strive for doses and frequency of medications that are well
susceptibilities are completed. Therapy may be altered as tolerated, TW monitoring during treatment that continues
needed once susceptibilities are known. The primary drugs to show no detectable Mtb shedding and drug levels that
used in elephants are isoniazid (INH), rifampin (RIF), exceed the minimum inhibitory concentration (MIC) of
pyrazinamide (PZA), ethambutol (ETH), and levofloxacin the cultured organism. Drug levels should be confirmed at
(LEVO). Another common veterinary fluoroquinolone, 6-month intervals or whenever there are changes in drug
enrofloxacin (ENRO) has been used to treat TB in elephants, selection or dosage (see Table 94.3).
but its effectiveness may be questionable based on research Treatment failures in elephants have occurred in a few
showing that Mtb organisms develop rapid resistance to its cases. Some of these were associated with individual elephant
active metabolite, ciprofloxacin.18 Four concurrently used intolerance to medications and development of resistance.
drugs are recommended, although in some cases, three Ongoing research into additional diagnostic techniques,
drugs will suffice. INH and RIF are recommended in all safer drug regimes, and Mtb epidemiology in elephants
elephant Mtb treatment regimes. INH causes early rapid will hopefully improve our ability to manage this disease.
TABLE
94.1 Elephant Mycobacterium tuberculosis Diagnostic Tests
Type of
Test Test Samples Needed Interpretation of Results Test Advantages Test Disadvantages Availability Comments
Culture Direct Trunk washes most A positive result from a living The only test that False negatives in Available from Culture is the gold
common elephant indicates active confirms active live animals due laboratories standard for
Other body fluids infection and shedding infection. Test is to technique, approved for diagnosis of Mtb
may be submitted at the time of sampling. 100% specific intermittent shedding, mycobacterial in elephants
for culture (semen, A positive result from a TW are lesion size and testing and ideally Trunk wash cultures
vaginal secretion, necropsy sample indicates noninvasive, location, and activity. with experience may also grow
ocular secretion, infection at the time of inexpensive Test has poor culturing NTM
mucus, lung lavage death, but not whether sensitivity elephant samples Although two cases
samples)35 the animal was shedding Slow turnaround for because TW of pulmonary
Postmortem samples or if the infection was results (10 days to 8 are often heavily disease in
of either fresh latent weeks) contaminated and elephants have
or frozen tissue, A negative result from a TW require elephant special techniques been associated
including suspicious living elephant indicates and staff to be are needed for with M. szulgai,36
lesions such as the elephant was not trained for sampling processing NTM grown in
granulomas or shedding at the time of Lung lavage requires a trunk wash
caseated lymph sampling. A negative extremely specialized do not raise
nodes, plus lung, result from a necropsy endoscopic zoonotic concerns
trachea, thoracic lesion suggests either equipment plus or necessitate
lymph nodes, that the infection was sedation treatment
salivary gland, and resolved (only dead
lower esophagus organisms present) or
sections. Body that the submitted lesion
fluids as described was not Mtb. qPCR
above should be performed to
confirm presence of Mtb
organisms
qPCR Direct Trunk washes from A positive result indicates Potentially highly Testing still in progress, Only one laboratory Potentially high
living elephant, Mtb organisms within specific thus sensitivity and available in the US specificity
other body fluids the sample, but does Can be done using specificity are not yet (NVSL) and sensitivity
and mucus not prove active infection same samples known Cost is less than $50 when done in
Postmortem samples because qPCR only sent for culture For TW, elephant and US No charge if combination
as described above confirms DNA presence Can be performed staff must be trained TW is submitted with TW culture,
of an organism, but not on formalin-fixed for sampling particularly with
its viability tissue new techniques
A negative result indicates being developed
no Mtb DNA was present to enhance the
in the samples tested sensitivity of PCR
for TW
Continued
CHAPTER 94 Elephant Mycobacteriosis: New Diagnostics and Management
667
TABLE
94.1 Elephant Mycobacterium tuberculosis Diagnostic Tests—cont’d
Type of
Test Test Samples Needed Interpretation of Results Test Advantages Test Disadvantages Availability Comments
Acid-fast staining Direct Fluid or mucus smear A positive result indicates Very rapid time for Very low sensitivity and Readily available.
668 SE C T I O N 19 Elephants and Rhinoceroses
CMI, Cell-mediated immunity; IFN-γ, interferon gamma; Mtb, Mycobacterium tuberculosis; NTM, nontubercular mycobacteria; TB, tuberculosis; TW, trunk wash, qPCR, real-time polymerase chain reaction.
CHAPTER 94 Elephant Mycobacteriosis: New Diagnostics and Management
669
670 SE C T I O N 19 Elephants and Rhinoceroses
TABLE 24–29
94.2 Suggested Drug Doses for the Treatment of Elephants With Mycobacterium tuberculosis
*INH has been associated with adverse effects in elephants and higher doses may need to be lowered. Determine dose using MIC of organism and elephant
tolerance. Some elephants are completely intolerant of INH. ENRO has been used in treatment of TB in elephants but may not be effective due to potential for
development of rapid resistance to ciprofloxacin.
†
No pharmacokinetic data are available for oral or rectal levofloxacin in elephants but have been published for oral enrofloxacin.37 Enrofloxacin metabolite,
ciprofloxacin is ineffective in killing Mtb organisms by development of rapid resistance and is not used in human TB treatment regimens.18,38
TABLE
94.3 Example of a Mycobacterium tuberculosis Treatment Protocol for an Asian Elephant (Elephas maximus)*
*Sample protocol only. Elephants have been treated with these doses and intervals but that does not imply success or tolerance of these medications and doses
in other elephants. This table’s treatment schedules have been used in elephants and are adapted from those recommended for the treatment of Mycobacterium
tuberculosis (Mtb) in humans.38
†
Elephant should receive weekly trunk wash (TW) testing by combined culture and real-time polymerase chain reaction (qPCR) for first 8–10 weeks of therapy.
It is recommended that detectable shedding of Mtb should cease during initial phase before proceeding to continuation phase.
‡
A fluoroquinolone may be used instead of or in conjunction with rifampin (RIF) based on susceptibilities and elephant’s compliance with taking oral medications.
ETH, Ethambutol; INH, isoniazid; LEVO, levofloxacin; PZA, pyrazinamide; RIF, rifampin.
Conclusions
Occupational and Public Health
Considerations Mtb infection is an important disease of the Asian elephant
in human care, with animal and human health consid-
Mtb is an important zoonotic disease with regulatory erations. Much remains to be learned about the disease.
responsibilities for the veterinarian and elephant holding However, with appropriate treatment, management of Mtb-
institution. Once a diagnosis is made, it is important that infected elephants can be accomplished while mitigating the
the proper authorities be notified in a timely manner. Deci- risks to human health.
sions about human contact exposure and use of proper
personal protective equipment should be made with the References
recommendations and oversight of local public health
authorities. Meticulous health record keeping is strongly 1. Feldman M, Isaza R, Prins C, et al: Point prevalence and inci-
advised to assist in documentation of the disease and assist- dence of Mycobacterium tuberculosis complex in captive elephants
ing public health authorities.5 in the United States, Vet Q 33:25–29, 2013.
CHAPTER 94 Elephant Mycobacteriosis: New Diagnostics and Management 671
2. Chandranaik B, Shivashankar BP, Umashankar KS, et al: Diseases Society of America: treatment of tuberculosis, Am J Resp
Mycobacterium tuberculosis infection in free-roaming wild Asian Crit Care Med 167:603, 2003.
elephant, Emerg Infect Dis 23:555–557, 2017. 21. Filippini P, Iona E, Piccaro G, et al: Activity of drug combina-
3. Obanda V, Poghon J, Yongo M, et al: First reported case of fatal tions against dormant Mycobacterium tuberculosis, Antimicrob
tuberculosis in a wild African elephant with past human-wildlife Agents Chemother 54:2712–2715, 2010.
contact, Epidemiol Infect 141(7):1476–1480, 2013. 22. Spigelman MK: New tuberculosis therapeutics: a growing pipe-
4. Perera BVP, Salgadu MA, Gunawardena GSPDS, et al: First line, J Infect Dis 196:S28–S34, 2007.
confirmed case of fatal tuberculosis in a wild Sri Lankan elephant, 23. Leibert E, Rom WN: New drugs and regimens for treatment of
Gajah 41:28–31, 2014. TB, Expert Rev Anti Infect Ther 8:801–813, 2010.
5. Stakeholders Task Force on Management & Research Priorities of 24. Maslow JN, Mikota SK, Zhu M, et al: Population pharmacoki-
Tuberculosis in Elephants. Recommendations for the diagnosis, treat- netics of isoniazid in the treatment of Mycobacterium tuberculosis
ment and management of tuberculosis (Mycobacterium tuberculosis) among Asian and African elephants (Elephas maximus and Lox-
in elephants in human care. Retrieved from http://www.aazv.org/ odonta africana), J Vet Pharm Ther 28:21–27, 2005.
resource/resmgr/Files/RecommendationsTBinElephants.pdf, 2017. 25. Maslow JN, Mikota SK, Zhu M, et al: Pharmacokinetics of
6. Heymann DL: Control of communicable diseases manual, Wash- ethambutol (EMB) in elephants, J Vet Pharm Ther 28:321–323,
ington, DC, 2008, American Public Health Association. 2005.
7. Murphree R, Warkentin JV, Dunn JR, et al: Elephant-to-human 26. Zhu M, Maslow JN, Mikota SK, et al: Population pharma-
transmission of tuberculosis, 2009, Emerg Infect Dis 17:366–371, cokinetics of pyrazinamide in elephants, J Vet Pharm Ther
2011. 28:403–409, 2005.
8. Simpson G, Zimmerman R, Shashkina E, et al: Mycobacterium 27. Peloquin CA, Maslow JN, Mikota SK, et al: Dose selection and
tuberculosis infection among Asian elephants in captivity, Emerg pharmacokinetics of rifampin in elephants for the treatment of
Infect Dis 23:513, 2017. tuberculosis, J Vet Pharm Ther 29:581–585, 2006.
9. Zlot A, Vines J, Nystrom L, et al: Diagnosis of tuberculosis 28. Brock AP, Isaza R, Egelund EF, et al: The pharmacokinetics of a
in three zoo elephants and a human contact—Oregon, 2013, single oral or rectal dose of concurrently administered isoniazid,
MMWR 64:1398–1402, 2016. rifampin, pyrazinamide, and ethambutol in Asian elephants
10. Lewerin SS, Olsson S-L, Eld K, et al: Outbreak of Mycobacterium (Elephas maximus), J Vet Pharm Ther 37:472–479, 2014.
tuberculosis infection among captive Asian elephants in a Swedish 29. Egelund EF, Isaza R, Alsultan A, et al: Isoniazid and rifampin
zoo, Vet Rec 156:171–175, 2005. pharmacokinetics in two Asian elephants (Elephas maximus)
11. Isaza R, Ketz C: A trunk wash technique for the diagnosis of infected with Mycobacterium tuberculosis, J Zoo Wild Med
tuberculosis in elephants, Verhandlung Internationalen Symp 47:868–871, 2016.
Erkrank Zootiere: 121–124, 1999. 30. Wiedner E, Schmitt DL: Preliminary report of side effects
12. Chambers M: Review of the diagnosis of tuberculosis in associated with drugs used in the treatment of tuberculosis in
non-bovid wildlife species using immunological methods–an elephants, Proceedings of the International Elephant Founda-
update of published work since 2009, Transboundary Emerg Dis tion, 2007, pp 15–20.
60:14–27, 2013. 31. Wilson E, Mikota S, Bradford JP, et al: Seropositive, culture
13. Greenwald R, Lyashchenko O, Esfandiari J, et al: Highly accurate negative tuberculosis in an Asian elephant (Elephas maximus),
antibody assays for early and rapid detection of tuberculosis in Proceedings of the American Association of Zoo Veterinarians/
African and Asian elephants, Clin Vaccine Immunol 16:605–612, American Association of Wildlife Veterinarians Joint Conference,
2009. 2010, p 170.
14. Angkawanish T, Morar D, Kooten PV, et al: The elephant inter- 32. de Steenwinkel JEM, de Knegt GJ, ten Kate MT, et al: Time–kill
feron gamma assay: a contribution to diagnosis of tuberculosis kinetics of anti-tuberculosis drugs, and emergence of resistance,
in elephants, Transboundary Emerg Dis 60:53–59, 2013. in relation to metabolic activity of Mycobacterium tuberculosis, J
15. Landolfi JA, Miller M, Maddox C, et al: Differences in immune Antimicrob Chemother 65:2582–2589, 2010.
cell function between tuberculosis positive and negative Asian 33. Egelund E, Alsultan A, Peloquin C: Optimizing the clinical
elephants, Tuberculosis 94:374–382, 2014. pharmacology of tuberculosis medications, Clin Pharm Ther
16. World Health Organization: 2011 commercial serodiagnostic 98:387–393, 2015.
tests for diagnosis of tuberculosis policy statement. Geneva, 34. Alsultan A, Peloquin CA: Therapeutic drug monitoring in the
Switzerland, 2011, p 26. treatment of tuberculosis: an update, Drugs 74:839–854, 2014.
17. Bryant JM, Grogono DM, Greaves D, et al: Whole-genome 35. Hildebrandt TB, Saragusty J, Moser I, et al: Bronchoalveolar
sequencing to identify transmission of Mycobacterium abscessus lavage technique: A new approach for diagnosis of tuberculosis
between patients with cystic fibrosis: a retrospective cohort study, infection in elephants, Proceedings of the American Association
Lancet 381:1551–1560, 2013. of Zoo Veterinarians/European Association of Zoo and Wildlife
18. Gumbo T, Louie A, Desziel M, et al: Pharmacodynamic evidence Veterinarians/Institute for Zoo and Wildlife Research, 2016, pp
that Ciprofloxacin failure against tuberculosis is not due to poor 113–114.
microbial kill but rapid emergence of resistance, Antimicrob 36. Lacasse C, Terio K, Kinsel MJ, et al: Two cases of atypical
Agents Chemother 49:3178–3181, 2005. mycobacteriosis caused by Mycobacterium szulgai associated with
19. Backues KA, Robbe-Austerman S, Isaza R: Documented ces- mortality in captive African elephants (Loxodonta africana), J Zoo
sation of mycobacterial shedding with antibiotic treatment in Wild Med 38:101–107, 2007.
a Mycobacteria-tuberculosis-positive Asian elephant (Elephas 37. Sanchez CR, Murray SZ, Isaza R, et al: Pharmacokinetics of a
maximus) by serial culture and direct real-time polymerase single dose of enrofloxacin administered orally to captive Asian
chain reaction testing of trunk wash samples, Proceedings of the elephants (Elephas maximus), Am J Vet Res 66:1948–1953, 2005.
American Association of Zoo Veterinarians, 2015, pp 151–152. 38. Nahid P, Dorman S, Alipanah N, et al: Official ATS/CDC/IDSA
20. Blumberg H, Burman W, Chaisson R, et al: American Thoracic practice guidelines: treatment of drug-susceptible tuberculosis,
Society/Centers for Disease Control and Prevention/Infectious Clin Infect Dis 63:e147–e195, 2016.
95
Elephant Endotheliotropic Herpesvirus
LAUREN LYNN HOWARD AND WILLEM SCHAFTENAAR
E
lephant endotheliotropic herpesvirus (EEHV) can have succumbed to EEHV-HD, and deaths have also been
cause acute, often fatal, hemorrhagic disease (EEHV documented in wild Asian elephants.12–15 EEHV is a global
hemorrhagic disease [EEHV-HD]) in young Asian issue that we are also fighting at a very local level.
elephants (Elephas maximus), most commonly between 1
and 8 years of age. For zoo veterinarians, this disease may be Asian Elephants
one of the most challenging and daunting aspects of caring
for a breeding elephant herd at their institution. The zoo Based on May 2017 population numbers, EEHV-HD was
clinicians’ best tools for managing EEHV are preparedness, the cause of 53% of deaths in all Asian elephants born in
vigilance, and an early, aggressive approach to treatment. North America since 1980, making it the single greatest
Details on the therapeutic approach to an elephant ill from cause of death in this cohort. The year 1980 is selected as a
EEHV-HD have been published, and this chapter is meant cutoff in North America because of the increased reliability
to augment, not replace, that important information.1 of records, and the beginning of captive births around that
time period. As of May 2017, there have been 35 cases of
EEHV-HD confirmed in Asian elephants born in North
Elephant Endotheliotropic Herpesvirus America since 1980 (of 141 elephants born), indicating
Impact and Epidemiology that 25%, or one in four, elephants in this cohort have
developed EEHV-HD. Eleven of these elephants survived
Healthy Asian and African (Loxodonta africana) elephants infection and 24 succumbed, leading to an overall mortality
have been shown to shed one or multiple types of their rate of 68% in North America.
species specific (or species-related) EEHV virus as part of In Europe, of more than 200 Asian elephants born since
a natural infection cycle that has evolved over millions 1995, 43 have died, and 60% of those deaths (26) were due
of years.2–6 Based on current research, all adult elephants to EEHV-HD, making it the largest cause of death in Asian
appear to carry and shed one or more EEHV strains and/ elephants born in Europe, as well.
or species intermittently and asymptomatically. Applying The first published, polymerase chain reaction (PCR)-
known herpesvirus biology to EEHV, this tells us that all confirmed case of EEHV in Asia was reported in Cambodia
adult elephants survived an episode of EEHV exposure in 2006.16 More recently, publications have documented a
and viremia at some point in their lives and are now latent fatal EEHV1 infection in Laos, fatal EEHV1 and EEHV4
carriers of the virus. infections in Thailand, and nine fatal EEHV1 infections
Viruses endemic within Asian and African elephant in both free-ranging (n = 4) and captive elephants (n = 5)
populations are listed in Table 95.1. The complete genome in India.12–14 Healthy Asian elephants under human care in
sequences have been determined for all the major EEHV India were shown to shed EEHV1, EEHV4, and EEHV5
species endemic for Asian elephants (EEHVs 1, 4, and from their trunks in 2014, a finding very similar to what
5).7–10 This achievement has led to significant advances in is seen in Asian elephant herds in North America and
the understanding of the evolution of EEHV and develop- Europe.4,3,17 Much remains to be learned about the impact
ment of tools with which to diagnose, treat, and develop of EEHV on the estimated 15,000 captive Asian elephants
future vaccines to EEHV.11 Although exposure to EEHV and greater than 40,000 wild elephants across Asia. During
appears to be a natural process, Asian elephants between 1 the First and Second Asian EEHV Strategy Meetings in
and 8 years of age are at high risk of developing EEHV-HD 2015 and 2016, more than 80 cases of EEHV-HD were
associated with EEHV infection, most commonly due to identified among most Asian elephant range countries
EEHV1.1 Older Asian elephants and African elephants are (Thailand, India, Cambodia, Indonesia, Myanmar, Nepal,
also susceptible to EEHV-HD but with less documented Sabah, Borneo, Laos, and Peninsular Malaysia), with less
frequency thus far. Asian elephants under human care in than 5 elephants surviving EEHV-HD and 12 of the
North America, Europe, and several Asian range countries deaths identified in free-ranging elephants in India. Wildlife
672
CHAPTER 95 Elephant Endotheliotropic Herpesvirus 673
TABLE Endemic Elephant Endotheliotropic most often used to treat EEHV, and the amount required to
95.1 Herpesviruses treat an elephant is not readily available on short notice. All
protocols should be developed jointly with veterinary and
Fatalities Year of elephant care teams, with support of key zoo administra-
Worldwide Discovery tors. Decision-making strategies and communication tactics
Asian Elephants (Elephas Maximus) should be discussed ahead of time so that critical time
EEHV1A 38 1999 is not wasted on long meetings when an elephant is ill.
Preparation for EEHV may be a costly and time-consuming
EEHV1B 4 2001
process and requires the full cooperation of all stakeholders.
EEHV4 2 2007 A potentially overlooked hallmark of EEHV preparedness
EEHV5 1 2008 is to cultivate and maintain open lines of communication
between the veterinarians and the elephant care team and
African Elephants (Loxodonta Africana)
to establish institutional support from administrators and
EEHV2 2 1999
public relations personnel. This is critical to allow for bilat-
EEHV3 1 2007 eral flow of important information, as well to streamline
EEHV6 1 2009 the process of discussions and decisions that will be part of
any EEHV-HD case.
EEHV7 0 2010
• BOX 95.1 Clinical Findings Associated With Elephant Endotheliotropic Herpesvirus Hemorrhagic Disease in
Pre, Early, and Peak Elephant Endotheliotropic Herpesvirus Viremia in Asian Elephants (Elephas
maximus)
each individual elephant, which will allow for identification of FCV against EEHV has not been proven. However, to
of subtle changes that may be a first clue to a more serious date, there are no peer-reviewed data available to establish
illness. Clinical findings in pre, early, and peak EEHV-HD that FCV does not have effect against EEHV. Until proven
viremia are listed in Box 95.1. Detailed hematologic and otherwise, it remains best practice to treat EEHV-HD cases
clinical findings in EEHV-HD are listed in Box 95.2. with FCV. Pharmacokinetic data of FCV in healthy Asian
The recommendation of the EEHV Advisory Group is elephants and limited data from clinically ill animals indi-
to monitor at-risk elephants (1–8-year-old Asian elephants) cate that potentially therapeutic blood levels of penciclovir
weekly for EEHV viremia via whole blood quantitative PCR may be achieved.28,30–32 Antivirals are only one aspect of
(qPCR). This is the best way to detect emerging EEHV-HD treatment, and it is becoming apparent that supportive care
early and allow for early, aggressive treatment. In addition, is just as important, if not more so, in the management of
measurement of CBC of at-risk elephants weekly helps to an EEHV-HD case.1,28,29 Rectal fluids should be initiated
establish individual reference ranges for key parameters immediately, and they have a striking ability to improve an
(white blood cell count, monocytes, and platelets) and elephant’s hydration and demeanor. Rectal fluids should be
helps us identify subtle decreases that may be associated administered through a soft-tipped hose and may be admin-
with early viremia. If at-risk elephants show any signs of istered as frequently as every 2 hours, at a minimum of 3–4
abnormal behavior, including decreased appetite, changes times daily in dehydrated animals. Fresh and frozen-thawed
in sleep patterns, lameness, or changes in mentation or elephant plasma, along with crystalloids, may be admin-
training, blood should be collected immediately for EEHV istered via intravenous boluses under standing sedation,
qPCR and CBC, even if this requires standing sedation to every 1–3 days as indicated by clinical condition and CBC
accomplish. status. Antibiotics for secondary infections, antiinflamma-
tories (at low doses in well-hydrated animals), and opioids
have also been given to elephants with EEHV-HD. A small
Early, Aggressive Treatment for Elephant number of Asian elephants ill from EEHV-HD have been
Endotheliotropic Herpesvirus treated with corticosteroids, when evidence of disseminated
intravascular coagulation is observed. This treatment option
All ill young elephants should be considered as possible has not been evaluated thoroughly enough to be strongly
EEHV-HD cases until proven otherwise by the results of recommended, although clinicians should consider its use.
whole blood qPCR testing. Treatment should be initiated Table 95.2 describes dosages and frequencies recommended
rapidly and often before confirmation of EEHV qPCR for EEHV therapy.
results is possible. FCV is the antiviral most often used in When to treat an elephant with EEHV-HD is as impor-
North America and in Europe to treat EEHV-HD, and tant as how to treat one. With weekly monitoring of CBCs
acyclovir and ganciclovir have also been used. The efficacy and EEHV qPCR in at-risk elephants, the zoo clinician
CHAPTER 95 Elephant Endotheliotropic Herpesvirus 675
• BOX 95.2 Detailed Clinical, Laboratory, and Physical Examination Findings by Group in Disease
Associated With Elephant Endotheliotropic Herpesvirus Hemorrhagic Disease in Asian
Elephants (Elephas maximus)
Clinical Observations Associated With EEHV-HD Laboratory and Physical Examination Findings
Group 1: Changes in Activity/Sleeping Pattern Associated With EEHV-HD
Sleeping more Group 1: Presence of EEHV Viremia
Sleeping less Confirmed via whole blood PCR at a validated EEHV testing
laboratory
Group 2: Signs of Discomfort or Acute Pain
Abdominal pain: stretching, rolling, colic behavior Group 2: Clinicopathologic Abnormalities (20%) Above
Musculoskeletal pain: lameness or stiffness, may shift between or Below Normal Values
legs >2% neutrophil bands present
Changes in WBC: leukopenia (early viremia) or leukocytosis
Group 3: Changes in Water or Food Intake, or in Fecal (later)
Output Changes in monocytes: monocytopenia (early viremia) or
Decreased appetite monocytosis (later)
Decreased water consumption Changes in platelets: thrombocytopenia (early viremia) or
Diarrhea or hard stools thrombocytosis (later)
Decreased stool production, constipation Changes in RBC: Anemia
Group 4: Mentation Change Elevations in acute phase proteins
Subdued, lethargic Group 3: Cardiac Abnormalities Noted on Physical
Appears confused, any neurologic signs Examination
Decreased participation in training with keepers Tachycardia
Group 5: Oral Lesions Arrhythmia
Hyperemic oral mucosa Heart murmur
Petechiae/ecchymoses Changes in blood pressure (must know individual baseline)
Pulse oximetry <95%
Group 6: Ocular Abnormalities
Scleral injection Group 4: Alteration in Body Temperature (Must Know
Icterus of sclera Individual Baseline)
Retinal hemorrhage Fecal bolus temperature: Above or below normal
may identify a case of EEHV-HD early, often prior to the information on this devastating disease. Full necropsy
onset of any visible clinical signs of illness. Alternatively, guidelines are available at www.eehvinfo.org.
regular monitoring of blood via qPCR may also pick up
low-level, subclinical viremia that is likely a normal occur- Future Directions
rence in young elephants. It may be a challenge to predict
if viremia will remain subclinical or will climb and cause Although our understanding of EEHV has grown astro-
clinical disease, and repeated blood testing, up to daily, may nomically in the past 5 years, there is still very much we
be necessary. Fig. 95.1 shows criteria for starting treatment do not know about this virus. Current research efforts
for EEHV-HD. Box 95.3 lists criteria for discontinuing are focused on antibody measurement and T-cell assays
EEHV-HD treatment. It is important to note that most to identify key immunogenic viral proteins as a basis for
of the data collected thus far are based on experience with future vaccine development and tools to evaluate response
EEHV1A and EEHV1B in Asian elephants, whereas inter- to such vaccines, as well as fine tuning EEHV-HD treat-
pretation of viral loads for EEHV4, EEHV3, and EEHV5 ment recommendations. Some headway is being made into
in Asian elephants and any viral loads in African elephants understanding the elephant immune response, although
are not as well established. there is much more to learn.33–35 An ultimate goal of the
Even conscientious monitoring and timely treatment EEHV community is to develop an EEHV vaccine to
cannot guarantee a successful outcome. Necropsy of an decrease the severity of clinical illness, if not eliminate illness
EEHV-HD case is an important opportunity to gain more altogether, and there is still much work to be done before
676 SE C T I O N 19 Elephants and Rhinoceroses
TABLE
95.2 Treatment Recommendations for Elephant Endotheliotropic Herpesvirus Hemorrhagic Disease
Recommendation Comments/Source
Fluid Therapy
Rectal fluids Bolus of 10–20 mL/kg TID to QID, up to Warm water, hose or tap water, administered with
every 2 h soft-tipped hose
Crystalloids IV bolus 0.3–4 mL/kg Should always be followed by rectal fluids*
†
Elephant plasma IV bolus 0.5–2 mL/kg
Antiviral Therapy
Famciclovir 15 mg/kg PO or rectally TID‡ Elephant PK30
Ganciclovir 5 mg/kg IV BID, each dose given slowly Used in 2 elephants in United States with no
diluted in 1 L of NaCl apparent adverse effects
Aciclovir 15 mg/kg BID orally, rectally or IV Used in 2 elephants in Thailand and 1 in Sweden
(survived EEHV-HD)
Antibacterial therapy Broad spectrum recommended See literature1
Adjunctive Therapy
Butorphanol 0.008–0.014 mg/kg IM q4h For signs of discomfort/colic
Omeprazole 0.7–1.4 mg/kg PO SID Equine dose36
Flunixin meglumine 0.25–0.5 mg/kg IM SID Equine antiendotoxemia dose,36 only in well-hydrated
animals
Furosemide 1 mg/kg IM Equine dose36
Triamcinolone 0.067 mg/kg IV Suggested dose, use only if signs of DIC are seen
Dexamethasone 0.05–0.1 mg/kg IV or IM Suggested dose, use only if signs of DIC are seen
Standing Sedation
Asian Calves
Butorphanol 0.045–0.075 mg/kg IM Houston Zoo EEHV Protocol
Detomidine 0.011–0.022 mg/kg IM Houston Zoo EEHV Protocol
Atipamezole 5 times detomidine dose Houston Zoo EEHV Protocol
Naltrexone 2.5–5 times butorphanol dose Houston Zoo EEHV Protocol
Light/Calming Sedation
*Asian elephants (Elephas maximus) have very low serum osmolarity and are hyponatremic and hypochloremic compared with other species. The normal serum
osmolarity range of Asian elephants is 252–270 mOsm/L. Commercial crystalloids should be considered hypertonic for elephants and should always be followed
by rectal fluid administration. (E. Wiedner, personal communication, May 2015.)
†
Plasma should be polymerase chain reaction (PCR) tested when collected from the donor elephant, and cross matching should be performed to reduce the
chance of transfusion reactions. More information on plasma transfusion and cross matching may be found on www.eehvinfo.org.
‡
Once viral load peaks and starts to decline, the author has decreased frequency to BID.
§
Reversal not always needed; cows usually recover smoothly on own.
DIC, Disseminated intravascular coagulation; EEHV, elephant endotheliotropic herpesvirus; EEHV-HD, elephant endotheliotropic herpesvirus hemorrhagic
disease; IM, intramuscular; IV, intravenous.
CHAPTER 95 Elephant Endotheliotropic Herpesvirus 677
No clinical signs, abnormal WBC4; Start daily monitoring qPCR and WBC, consider
Viral load + but <5000 VGE/mL starting first phase of EEHV treatment
Clinical signs,5 normal WBC; • Start first step of EEHV treatment: antivirals,
Viral load: unknown rectal fluids TID-QID
• Additional diagnostic examinations to rule
out non-EEHV related disease
Clinical signs, abnormal WBC;
Viral load + but <5000 VGE/mL • Start daily monitoring qPCR and WBC
1
Normal WBC: platelets and monocytes within normal ranges of the elephant concerned, compared to individual historical ranges
2
Duration between recheck samples is dependent on proximity of EEHV qPCR laboratory and turnaround time for qPCR and
CBC test results
3
VGE = viral genome equivalents
4
Abnormal WBC: thrombocytopenia and/or monocytopenia and/or leucopenia, compared to individual historical ranges
5
Clinical signs
6
DIC-symptoms: ptechiae, hemorrhages, sclera injection, edema, cyanosis, hydropericard (ultrasonograpy)
• Figure 95.1 Elephant endotheliotropic herpesvirus treatment in Asian elephants between 1 and 8
years of age. CBC, Complete blood count; DIC, disseminated intravascular coagulation; EEHV, elephant
endotheliotropic herpesvirus; EEHV-HD, elephant endotheliotropic herpesvirus hemorrhagic disease; IV,
intravenous; qPCR, quantitative polymerase chain reaction; WBC, white blood cell.
678 SE C T I O N 19 Elephants and Rhinoceroses
• BOX 95.3 Discontinuation of Elephant 5. Zong JC, Latimer EM, Long SY, et al: Comparative genomic
analysis of four elephant endotheliotropic herpesviruses, EEHV3,
Endotheliotropic Herpesvirus
EEHV4, EEHV5, and EEHV6, from cases of hemorrhagic
Hemorrhagic Disease Treatment in disease or viremia, J Virol 88:13547–13569, 2014.
Asian Elephants (Elephas maximus) 6. Long SY, Latimer EM, Hayward GS: Review of elephant endoth-
1–8 Years Old eliotropic herpesviruses and acute hemorrhagic disease, Inst Lab
Consider decreasing frequency of rectal fluids/famciclovir to BID
Anim Res 56:283–296, 2015.
when: 7. Ling PD, Reid JG, Qin X, et al: Complete genome sequence
• Viral load begins to decrease consistently (may bounce up of elephant endotheliotropic herpesvirus 1A, Genome Announc
and down first) 1(2):e00106-13, 2013, doi:10.1128/genomeA.00106-13.
• WBC/monocytes/platelets are normal or elevated 8. Wilkie GS, Davison AJ, Watson M, et al: Complete genome
• Elephant appears clinically normal sequences of elephant endotheliotropic herpesviruses 1A and
Consider discontinuing treatment with rectal fluids/famciclovir 1B determined directly from fatal cases, J Virol 87:6700–6712,
when: 2013.
• Viral load is <5000 vge/mL 9. Wilkie GS, Davison AJ, Kerr K, et al: First fatality associated with
• WBC/monocytes/platelets are normal or elevated
elephant endotheliotropic herpesvirus 5 in an Asian elephant:
• Elephant appears clinically normal
pathological findings and complete viral genome sequence.
WBC, White blood cell. Nature Scientific Reports, 2014. doi:10.1038/srep06299.
10. Ling PD, Long SY, Fuery A, et al: Complete genome sequence
of elephant endotheliotropic herpesvirus 4, the first example
of a GC-rich proboscivirus, mSphere 1(3):e00081-15, 2016,
that is accomplished. In addition, a better evaluation of the doi:10.1128/mSphere.00081-15.
epidemiology and distribution of EEHV in North America, 11. Ling PD, Long SY, Zong JC, et al: Comparison of the gene
Europe, and Asian range countries is important to better coding contents and other unusual features of the GC-rich and
understand the impact of this disease on wild and captive AT-rich branch probosciviruses, mSphere 1(3):e00091-16, 2016,
elephant populations. Capacity building, in the form of doi:10.11128/mSphere.00091-16.
EEHV diagnostic laboratories throughout Asia, is the first 12. Bouchard B, Xaymountry B, Thongtip N, et al: First reported
case of elephant endotheliotropic herpesvirus infection in Laos,
step in this process. Finally, education of zoo professionals,
J Zoo Wildl Med 45:704–707, 2014.
as well as the lay public, by sharing our success and advances 13. Sripiboon S, Tankaew P, Lungka G, et al: The occurrence of
with EEHV, is critical to establishing public support for elephant endotheliotropic herpesvirus in captive Asian elephants
elephant institutions and EEHV-related research. (Elephas maixmus): first case of EEHV4 in Asia, J Zoo Wildl Med
44:100–104, 2013.
Acknowledgments 14. Zachariah A, Zong JC, Long SY, et al: Fatal herpesvirus hemor-
rhagic disease in wild and orphan Asian elephants in Southern
The authors thank Erin Latimer, Dr. Paul Ling, and Dr. India, J Zoo Wildl Med 49:381–393, 2013.
Gary Hayward for their contributions to EEHV research 15. Kendall R, Howard L, Masters N, et al: The impact of elephant
and editing of this chapter. We also acknowledge Dr. Ellen endotheliotropic herpesvirus on the captive Asian elephant
Wiedner for her contributions to the elephant and EEHV (Elephas maximus) population of the United Kingdom and
Ireland (1995-2013), J Zoo Wildl Med 47:405–418, 2016.
community.
16. Reid CE, Hildebrandt TB, Marx N, et al: Endotheliotropic
EEHV infection, the first PCR confirmed fatal case in Asia, Vet
References Q 28:61–64, 2006.
17. Hardman K, Dastjerdi A, Gurrala R, et al: Detection of EEHV
1. Wiedner E, Howard L, Isaza R: Treatment of elephant endo- type 1 in asymptomatic elephants using TaqMan real-time PCR,
theliotrophic herpesvirus (EEHV). In Fowler ME, Miller RE, Vet Rec 170:205–208, 2012.
editors: Zoo and wild animal medicine, ed 7, St. Louis, 2012, 18. Latimer E, Zong JC, Heaggans SY, et al: Detection and evalua-
Elsevier, pp 537–543. tion of novel herpesviruses in routine and pathological samples
2. Richman LK, Zong JC, Latimer EM, et al: Elephant endoth- from Asian and African elephants: identification of two new
eliotropic herpesviruses of EEHV1A, EEHV1B, and EEHV2 probosciviruses (EEHV5 and EEHV6) and two new gammaher-
from cases of hemorrhagic disease are highly diverged from other pesviruses (EGHV3B and EGHV5), Vet Microbiol 147:28–41,
mammalian herpesviruses and may form a new subfamily, J Virol 2011.
88:13523–13546, 2014. 19. Pearson VR, Robbins H, Douglas-Hamilton I, et al: Elephant
3. Stanton JJ, Zong JC, Latimer E, et al: Detection of pathogenic herpesviruses EEHV2, EEHV3A, EEHV3B (a new subspecies),
elephant endotheliotropic herpesvirus in routine trunk washes EEHV6, EEHV7A, EEHV7B (a new subspecies) and EGHV1A,
from healthy adult Asian elephants (Elephas maximus) by use of EGHV1B (a new species), EGHV2, EGHV4 found in tissue
a real-time quantitative polymerase chain reaction assay, Am J Vet biopsies and saliva from African elephants in Kenya and America,
Res 71:925–933, 2010. in Proceedings of the International Elephant and Rhinocerous
4. Stanton JJ, Nofs SA, Zachariah A, et al: Detection of elephant Conservation and Research Symposium, 2013, pp 643–677.
endotheliotropic herpesvirus infection among healthy Asian 20. Richman LK, Montali RJ, Garber RL, et al: Novel endothelio-
elephants (Elephas maximus) in South India, J Zoo Wildl Med tropic herpesvirus fatal for Asian and African elephants, Science
50:279–287, 2014. 283:1171–1176, 1999.
CHAPTER 95 Elephant Endotheliotropic Herpesvirus 679
21. Richman LK, Hayward GS: Elephant herpesviruses. In Fowler 29. Fuery A, Tan J, Peng R, et al: Clinical infection of two captive
ME, Miller RE, editors: Zoo and wild animal medicine, ed 7, St. Asian elephants (Elephas maximus) with elephant endotheliotro-
Louis, 2012, Elsevier, pp 496–502. pic herpesvirus 1B, J Zoo Wildl Med 47:319–324, 2016.
22. Stanton J, Zong JC, Eng C, et al: Kinetics of viral loads and 30. Brock AP, Isaza R, Hunter RP, et al: Estimates of the pharma-
genotypic analysis of elephant endotheliotropic herpesvirus-1 cokinetics of famciclovir and its active metabolite penciclovir
infection in captive Asian elephants, J Zoo Wildl Med 44:42–54, in young Asian elephants (Elephas maximus), Am J Vet Res
2013. 73:1996–2000, 2012.
23. Zong JC, Heaggans SY, Long SY, et al: Detection of quiescent 31. Howard LL: Penciclovir levels in Asian elephants undergoing
infections with multiple EEHVs, including EEHV2, EEHV3, famciclovir treatment for clinical EEHV viremia at the Houston
EEHV6, and EEHV7, within lymphoid lung nodules or lung Zoo, in Proceedings of the 10th International Workshop for
and spleen tissue samples from five asymptomatic adult African EEHV, 2015, pp 33–34.
elephants, J Virol 90:3028–3043, 2016. 32. Schmitt DL, Hardy DA, Montali RJ, et al: Use of famciclovir
24. Bronson E, McClure M, Sohl J, et al: Epidemiologic evaluation for the treatment of endotheliotropic herpesvirus infections in
of endotheliotropic herpesvirus 3B (EEHV3B) infection in an Asian elephants (Elephas maximus), J Zoo Wildl Med 31:518–522,
African elephant (Loxodonta africana), J Zoo Wildl Med, 2017. 2000.
25. Kongmakee P, Suttiyaporn S, Changpetch W, et al: Elephant 33. Fuery A, Rooney CM, Makedonas G, et al: Detecting EEHV-
endotheliotropic herpesvirus type 6 infection in a captive African specific T cell responses in Asian elephants, in Proceedings of the
elephant (Loxodonta africana) in Thailand. In Proceedings of the 10th International Workshop for EEHV, 2015, p 13.
10th International Workshop for EEHV, 2015, p 41. 34. Humphreys AF, Tan J, Peng RS, et al: Generation and character-
26. Nofs SA, Hicks J, Ling P: Detection of EEHV in trunk wash ization of antibodies against Asian elephant (Elephas maximus)
secretions from captive African elephants (Loxodonta africana), in IgG, IgM, and IgA, PLoS ONE 10(2):e0116318, 2015.
Proceedings of the Annual Conference of the American Associa- 35. Van den Doel P, Prieto VR, van Rossum-Fikkert SE, et al: A
tion of Zoo Veterinarians, 2013, p 56. novel antigen capture ELISA for the specific detection of IgG
27. Atkins L, Zong JC, Tan J, et al: Elephant endotheliotropic antibodies to elephant endotheliotropic herpesvirus, BMC Vet
herpesvirus 5, a newly recognized elephant herpesvirus associated Res 11:203–212, 2015.
with clinical and subclinical infections in captive Asian elephants, 36. Plumb DC: Veterinary drug handbook, ed 8, Ames, IA, 2015,
J Zoo Wildl Med 44:136–143, 2013. Wiley-Blackwell.
28. Fuery A, Browning GR, Tan J, et al: Clinical infection of captive
Asian elephants (Elephas maximus) with elephant endotheliotro-
pic herpesvirus 4, J Zoo Wildl Med 47:311–318, 2016.
96
Elephant Pregnancy and Parturition:
Normal and Abnormal
IMKE LÜEDERS
A
lthough elephant births occur with some regularity in metabolites. The normal luteal phase is between 8 and 10
selected zoological facilities, globally, we still remain weeks; thus prolonged elevation of progestagens beyond
a long way from having a self-sustaining captive pop- 16 weeks is indicative of pregnancy. It should be noted,
ulation. Although the birthing process and postnatal period though, that a prolonged luteal life span has been observed
are critical periods for mother and calf, the increased number in nonpregnant Asian elephants as well as very low pro-
of captive elephant births during the past 20 years has led gesterone values in pregnant elephants (unpublished data).
to a greater understanding of elephant reproductive phys- However, the typical progestagen profile of a pregnancy
iology as well as to significant improvements in elephant shows an increase within a few hours up to 3 days postovu-
birth management. The aim of this chapter is to provide lation, after which levels rise and remain elevated for about
a general overview of elephant pregnancy, parturition, and 6–8 weeks. This is usually followed by a transient decline
peripartum complications and to make recommendations from weeks 8–10, before progestagens increase to pregnancy
regarding appropriate veterinary care for both the elephant levels. African and Asian elephants show slight differences
cow and calf when complications arise. Although Asian in their pregnancy hormone patterns.5
(Elephas maximus) and African elephant (Loxodonta afri- Pregnant and cycling elephants show multiple corpora
cana) reproductive physiology is similar during gestation lutea on both ovaries, despite being uniparous although
and parturition, a few differences exist, which are described. with rare twin births. A single corpus luteum that forms
In addition, more data have been generated on Asian ele- after ovulation develops along with additional, up to 10,
phants, because the numbers of captive births in African accessory corpora lutea (CL).3,6 These accessory CL are
elephants remain much lower than in Asian elephants, as is derived from luteinized follicles that appear after the first
similarly reflected in the ensuing text. of two luteinizing hormone peaks and prior to ovulation.6
The main steroid produced by elephant luteal cells is not
The Normal Pregnancy progesterone itself but its 5α-reduced metabolites.7 Thus
commercial progesterone assays must be tested and vali-
The mean gestation length (GL) of African and Asian dated for sufficient cross-reactivity. Serum prolactin (PRL)
elephants in European facilities is 642 and 655 days, measurements may provide another endocrine verification
respectively.1 Similar mean gestation periods of 640 and of pregnancy. Prolactin values increase by more than 100
658 days for African and Asian elephants, respectively, have times after months 4–6 of gestation and remain high until
been recorded in a study including gestations from North after parturition.8 Therefore a single elevated serum PRL
America and Japan.2 Thus Asian elephants carry longer than measurement 6 months after ovulation is indicative of
African elephants. No difference was seen between GL with pregnancy. Again, elephant PRL detection warrants special
respect to calf gender, but individual variation in GL is endocrine assays. Mating observations are not always reli-
high (Table 96.1), and GL appears to increase with parity able indicators because some bulls will still copulate with a
(A. Oerke, personal communication, November 3, 2016). pregnant female.
The elephant pregnancy is likely maintained only by Pregnancy confirmation is also possible by transrectal
progestagen secretion of multiple corpora lutea, because ultrasound, using a 2- to 7-MHz convex ultrasound probe.
the placenta is steroidogenically inert.3 Based on ultrasono- The earliest observations of an embryonic vesicle—a small,
graphic findings, implantation does not occur earlier than anechoic, 5–8 mm round structure within the uterine horn
45 days postovulation, after which luteal rescue occurs.3,4 lumen—may be possible by 45–50 days postovulation, but
Pregnancy is best confirmed by weekly serum hormone an inexperienced investigator should wait 70–100 days to
monitoring or noninvasively by urine or fecal progesterone confirm a true embryo.9,10 Transrectal ultrasound may be
680
CHAPTER 96 Elephant Pregnancy and Parturition: Normal and Abnormal 681
TABLE
96.1 Data Documenting Normal Elephant Pregnancies and Parturition Resulting in Live Births
African Elephant
Variable Asian Elephant (Elephas maximus) (n) (Loxodonta africana) (n)
General Pregnancy
GL total (range) 622–693 (28)1 624–660 (18)1
617–690 (21)2 625–667 (28)2
GL female calf (mean) 655 (10)1 646 (10)1
645 (11)2 654 (12)2
GL male calf (mean) 657 (18)1 638 (8)1
635 (10)2 661 (16)2
Beginning of breast development ~80% in 2nd or 3rd trimester (14)11
Beginning of milk secretion ~60% a few days prior parturition, 30% at 2–4
(prev. nonlactating females) weeks prior parturition, seldom cases of
several months prior birth (10)11
Labor
Time from serum progesterone drop <24 h up to 7 days, usually within 1–3 days
until parturition (personal observation),
up to 14 days reported15
Time from expulsion of mucus plug ~ 60% within 12 h, up to 72 h recorded (14)11
until visible labor
Time from expulsion of mucus plug median: 15 h, range:
until parturition 4–99 h (8)28
Time from first active labor until calf ~70% within 6 h, up to 24 h recorded (25)11
expulsion
Time from bulge under tail until median 13 min, range:
parturition 3 min–2.5 h (10)28
Parturition
Fetal position during birth: 65% (37)11–71% (35)20
Posterior presentation
Fetal position during birth: 29% (35)20–35 % (37)11
Anterior presentation
Time from rupture of allantois until 90% within min to 6 h, one case of 55 h (32)11
birth
Afterbirth expelled 85% within 6 h (41)11 median: 3 h, range:
72 min–17 h (12)28
Because more captive births of Asian elephants have been recorded, most data refer to this species. It should be noted that large variation exists between
individual elephants. Thus deviations from these values may still result in healthy calves being born.
n, Number of subjects contributing; GL, gestation length.
• Figure 96.1 Schematic diagram of a pregnant Asian elephant giving birth to a fetus in posterior
position. Usually the allantoic sac breaks, releasing fluids, and the calf slides fully enveloped within the
amniotic membranes through the vagina and vaginal vestibulum.
The normal progress of events during parturition may months (see Table 96.1). Prediction of parturition is possible
be described as follows: (1) Onset of increased discomfort on an individual basis only by observation of physiologic or
7 days to 48 hours prior to birth; (2) restlessness and first behavioral changes or, preferred, daily serum progesterone
abdominal contractions representing the first stage of labor; determinations, because progestagen levels usually drop at
(3) loss of the mucus plug that sealed the vagina during least below two-thirds of the mean pregnancy value a few
pregnancy; (4) second stage of labor with increased signs of days prior to parturition (Fig. 96.2).13,14
pain, stronger abdominal contractions, straining, agitation; Physical changes predictive of birth may not always be
(5) rupture of allantois, although water may break later; as obvious in elephants as in other species, especially if the
(6) appearance and disappearance of a bulge under the pregnant female has a nursing calf with her, which precludes
tail below the anus and advancement of the fetal feet (see use of breast development and milk secretion as indicators.
Fig. 96.1); (7) third stage of labor with strong contractions Ventral abdominal or vulval edema may become visible
and final entrance of the fetus into the vestibulum; (8) early in gestation and may cause discomfort, although
continued visibility of the bulge as soon as the calf ’s body edema has not been observed to cause any difficulties during
enters the female’s pelvic cavity (and the hymen breaks in parturition. Regular exercise and hydrotherapy with cold
nulliparous cows); (9) movement of the calf through the and warm water may be helpful to increase circulation and
vaginal vestibulum and expulsion from the mother’s body, avoid the development of necrotizing tissue.
usually still covered in the bluish amnion; (10) delivery of One indication of upcoming birth is loss of the mucus
the afterbirth (girdle placenta and allantoic membranes) 45 plug from the vaginal vestibulum, which usually occurs a
minutes to a few hours later. few hours prior to birth, although up to several days prior
The stages are not always clearly discernible from each or even partial mucus plug loss in the middle of gestation
other, and large variation in time spans from what is have also been described (see Table 96.1).14,15 The plug can
described here may still result in a successful birth (see be white, yellowish to brown, sticky and mucoid, or bloody.
Table 96.1). The entire process may be finished within less Other signs may include frequent defecation and urination,
than an hour but may take up to 48 hours. small fecal ball size, reduced appetite, restlessness, nervous-
ness, beating the vulva with the tail, and finally labor. Onset
Prediction of Parturition of the labor is characterized by “freezing” intermittently
(standing still with no movement), spread legs, moving
The most likely time of parturition can be calculated as flanks, tail flagging, straining, kneeling, lateral recumbency,
620–660 days from the day of last observed mating or mea- and repeatedly lying down and then standing back up.
sured rise in progestagens. As described earlier, gestational The most reliable and exact measurable indicator for
length in elephants resulting in live birth can vary by up to 3 imminent parturition is through the daily measurement of
CHAPTER 96 Elephant Pregnancy and Parturition: Normal and Abnormal 683
2,0
La1 – 2013
La2 – 2007
1,8
La2 – 2011
La2 – 2016
1,6
La3 – 2008
La3 – 2014
1,4
Em1 – 2015
Serum progesterone [ng/ml]
1,2
1,0
0,8
Parturition
0,6
0,4
0,2
0,0
13 12 11 10 9 8 7 6 5 4 3 2 1 0
Day until parturition
• Figure 96.2 Examples of hospital-measured serum progesterone courses at the end of pregnancy
in three different African and one Asian elephant with live-born calves (La1: one pregnancy, La2: three
pregnancies, La3: two pregnancies; Em1: one pregnancy). Daily and later twice-daily serum samples
should be measured to detect imminent parturition in elephants. Hormone values start decreasing
significantly up to 5 days prior to parturition (day 0), but as late as 1 day prior to giving birth in these
examples. (Data credit: Dr. Arne Lawrenz, Wuppertal Zoo, Germany.)
A B
• Figure 96.3 Asian elephant ultrasound images of the cervix and uterine body during parturition. The
images were obtained when labor stopped for more than 2 hours. (A) When allantoic (al) and amniotic
membranes (am) were seen pushing into the opened cervix and (B) during labor, fetal feet (ff) were
observed, the decision was made to administer 40 IU of oxytocin intramuscularly, and the calf was born
30 minutes later. (Image credit: Dr. Arne Lawrenz, Wuppertal Zoo, Germany.)
fetus may remain within the uterus or be naturally expelled relationship between obesity in zoo elephants and dystocia thus
days to years after death.15,18,19 appears likely.15
Malposition/Malformation of Fetus
Complications During Parturition
Dystocia has been linked to both breech position and
Problems during parturition in captive elephants occur rela- anterior presentation.15,18,20 In a recent study, 60% of calves
tively often and may be fatal for mother and/or calf. Recent born head first experienced dystocia, compared with 16%
statistics from the European population indicate that 12% born hindlimb first.20 The same study found that 40% of
of calves (Asian and African) are stillborn.20 However, Asian the calves born head first were stillborn compared with
timber elephants in Myanmar reportedly experience only only 12% of those born hindlimbs first. Dystocia is report-
4% stillbirths.12 edly associated with deformities that include ankylosis,
Overall, Asian elephants seem to be more prone to dys- encephalomeningocoele, hydrocephalus, spina bifida, cleft
tocia compared with the African species.19 In both species, palate, missing maxilla and trunk, and tetralogy of Fallot,
dystocia is significantly associated with stillbirth, resulting or with malpositions such as head tilt, twisted legs, and
in a 50% chance in African elephants and 86% chance in entanglement with the umbilical cord.15,22
Asian elephants of a dead-delivered calf.20 Dystocia in Asian
elephants was more often reported in historic data from Weak Labor/Uterine Inertia
Europe, with an incidence rate of 36%,11 compared with Dystocia has furthermore been linked to lack of physical
only 16.4% of Asian elephants in a more recent study.20 fitness,2 calcium deficiency,23 and insufficient labor due
to external stressors.11,15,19 Studies suggest that calcium
Reasons for Dystocia levels may actually be too low in general, and the authors
have reported a dystocic Asian cow whose calcium was at
A number of factors may lead to dystocia and subsequent baseline.23 Normal plasma concentrations of total calcium
stillbirth or sometimes retention of the fetus. These are should be around 3.6 mmol/L, and normal plasma concen-
discussed in the following paragraphs. trations of ionized calcium should be around 1.25 mmol/L.
Elephants should be fed calcium- and cholecalciferol-rich
Oversized Fetus diets at all times, particularly when close to parturition.
More problems during parturition seem to occur in the Additionally, serum calcium levels should be monitored
Asian elephant species compared with African elephants. closely prior to parturition.23
The reason may be that Asian elephant calves exceed Africans It is believed that external stress factors may lead to a delay
for mean birth height (93.7 vs. 89.9 cm) and birth weight in the birthing process. Some authors suggest that elephants
(116.5 vs. 103.5 kg).2 Stillborn Asian elephant calves are may actively interrupt labor, especially when humans are
heavier compared with live-born calves.2,20 present.11,15,19 However, this is difficult to confirm. Labor
The calf ’s body weight may be related to preg- is a time of psychophysiologic stress. In humans, it is
nancy length and the body condition of the female. A well established that cortisol levels increase throughout
CHAPTER 96 Elephant Pregnancy and Parturition: Normal and Abnormal 685
pregnancy and continue to increase at term with advancing valuable tools are available for assessing the situation and
labor.24 These physiologic changes are important and may taking appropriate actions. Use of rectal palpation and, even
be a necessity for maintaining maternal/fetal well-being and better, transrectal ultrasonography are the first steps to judge
for promoting normal labor progression.24 the situation. The clinician should determine the position of
For wild elephants, allomothering and herd-supported the fetus and whether any body parts have entered the birth
births are the norm. However, many captive elephant herds canal. This requires an elephant cow, whether in restricted
still consist of unrelated individuals. A subdominant female or nonrestricted contact, to be trained to allow these
with no relationship to the rest of the herd may be stressed procedures on a routine basis. Remote camera monitoring
by feeling vulnerable and intimidated by other elephants. may help to generate information on behavior and crucial
Thus being separated from the group might relieve stress, events such as time of onset of labor or allantoic rupture.
and tethering might be beneficial for an inexperienced For elephants managed in a captive breeding program, it is
female. However, in another elephant unused to restraint, furthermore advised to contact the Taxon Advisory Group
tethering and separation might increase stress. Survey data veterinary advisors or colleagues experienced with elephant
suggest separation from other elephants and chaining as birth management.
high-risk factors for dystocia.12 Regardless, preplanning,
preparation, and habituation to the envisaged birthing Conservative Approaches for Fetal Delivery
situation are essential to reducing external stressors for an
expectant elephant. If the fetus is not delivered despite an open birth canal due
to weak labor or if the birthing process stops, the following
Difficulties in Passage Through the Birth Canal steps may be taken.
Inadequate dilation of the cervix, poor softening of the
vagina and the vaginal vestibulum in older, primiparous Triggering the Ferguson Reflex
females; fibrosis of the hymen; lower reproductive tract Before any medical stimulation of labor and ripening of the
pathologies; and obesity may all pose obstacles during cervix, manual massage of the rectal floor may be attempted
parturition.2,15 Although older survey data suggest that nul- to stimulate the underlying vagina and cervix. Again, train-
liparous cows more than 30 years of age experience a higher ing the elephant to allow rectal palpation and manipulation
incidence of stillbirth and dystocia,25 more recent surveys is essential. Both arms up to the elbows may be inserted
do not support an association between age and dystocia but into the elephant’s rectum. By repeatedly pulling one’s
rather with primiparity.20 Thus parity is a primary factor for knuckles along the bottom of the rectal wall toward the anus
passage problems. with some force, stimulation of underlying vaginocervical
Fitness and physical condition may play a role in the receptors will trigger release of endogenous oxytocin (the
successful expulsion of a calf as heavy as 100–140 kg. This Ferguson reflex), helping relax the cervix and induce uterine
may explain why there is only a 4% stillbirth risk in working smooth muscle contractions.15
Myanmar timber elephants compared with a 12% risk in
zoo elephants.21 Exercise does help with weight loss and Calcium
body shape and thus may increase the elephant’s overall If weak labor is observed and the normal passage through
physical fitness. the birth canal seems possible, serum calcium should be
In Myanmar timber elephants, stillbirth probability measured. If total and ionized calcium is below 2.5 and
dropped from 11.3% for first-born calves to only 1.8% 1.2 mmol/L, respectively, oral or a slow intravenous (IV)
for later-born calves. Thus first-born calves were 6.83 times infusion of calcium should be administered.15,23 Intra-
more likely to be stillborn than subsequent calves from the venous administration of Ca-gluconate (23%) at a dose
same mother.21 This has also been confirmed for captive of 400–2000 mL has been reported in a dystocic Asian
Asian elephants in Europe: of 41 first-born calves, 11 were elephant.26
stillborn (26.8%); but of 116 later-born calves, only 6 were
stillborn (5.2%).11 Oxytocin
The hymen-like structure that “seals” the entrance The elephant uterus appears to be very sensitive to oxytocin.
from the vaginal vestibulum into the vagina breaks only Before this drug is administered, the degree of cervical
during first parturition (not during mating). The hymen dilation must be assessed. If the cow is multiparous, the
may become increasingly fibrotic with age and thus cervix is open, parts of the fetus have entered the vagina,
present another obstacle during birth in older first-time and the birthing process has stopped for at least an hour,
mothers.15 oxytocin administration may be considered.15 Uterine
rupture has occurred in some cases when these precondi-
Obstetrics tions were not met. The dose recommendations range from
20–60 IU intramuscularly (IM) every 2 hours (personal
Veterinary intervention options are limited in elephants due experience).15 Note that it may take as long as 20 minutes
to their size and special reproductive anatomy. Nevertheless, from IM injection until the onset of full effect. If avail-
in accessible elephants with a high level of training, several able, carbetocin (e.g., Depotocin, 70 µg/mL, Veyx-Pharma
686 SE C T I O N 19 Elephants and Rhinoceroses
Denaverine
If the cervix is not dilated and possibly only one of the
calf ’s legs has entered the vagina, besides manual stimu-
lation of the Ferguson reflex, denaverine hydrochloride
(Sensiblex, 40 mg/mL Veyx-Pharma GmbH, Germany)
where available may provide an option. This antispas-
modic drug is used in domestic cattle and dogs and acts
to help relax the cervix and the rest of the birth canal
while not affecting labor. There is not much experience
with its use in elephants, but it may be worth considering
in nulliparous females with suspected inadequate opening
of the cervix. This drug is administered IM at a dose of • Figure 96.4 Episiotomy in a dystocic Asian elephant with subse-
800–1600 mg, which may be repeated once an hour later. quent fetotomy. Chains have been attached to the fetal feet through
the incision. After successful attachment of the chains to a part of the
It should be noted that this drug has no effect on early dead fetus, the chain was guided through the distal vaginal vestibulum
cervical dilation. and the dissected parts of the fetus were removed, following the
natural route to avoid tissue trauma at the incision site. (Photo credit:
Estradiol and Prostaglandin E Dr. Willem Schaftenaar, Rotterdam Zoo, The Netherlands.)
Other drugs recommended in elephants to induce dilation
of the cervix are transrectal or transcutaneous application of
estradiol or prostaglandin E.13 Estradiol gel (600–800 mg)
may be applied by transrectal massage and may be repeated access to the vagina and cervix is possible (Fig. 96.4). Thus,
at a dose of 300–400 mg every 3–4 hours.15 Both prosta- to reach and remove a dead fetus lying within the cervix
glandin E1 (PGE1) and prostaglandin E2 (PGE2) are used and vagina, direct access into the vaginal vestibulum must
in humans and mares for cervical ripening and induction be established by a means of a vertical incision of about
of labor and may be helpful in elephants. Misoprostol, 15 cm length, 8 cm ventral to the anus (see Fig. 96.4).15,22
a synthetic PGE1, has been administered to elephants to Extraction forces should be applied physiologically (i.e., by
dilate the cervix and induce uterine contractions at empiric pulling ropes or chains attached to the fetal legs or trunk
doses of approximately 1000 mg orally or 500 mg transrec- in the direction of the ground, e.g., by guiding the ropes
tally every 12 hours for a 4000-kg elephant.15 Administra- through metal rings fixed at the floor) and not horizontally.
tion of dinoprostone, a PGE2 analogue, used at a dose Although use of local anesthesia rather than standing seda-
of 1.5–2.5 mg transrectally above the cervical region, was tion has been recommended to enable the cow’s straining
suggested to be effective.15 to assist in fetal expulsion,15 this may not always be feasible,
as some females will require chemical restraint to tolerate
Surgical Approaches for Fetal Delivery the procedures. In cases of fetal oversize, malposition, or
malformation, fetotomy is a last option; but it is needed
Surgical management of dystocia also warrants thorough because leaving a fetus within the birth canal will result
assessment of the situation first. Only if the calf has entered in fatal ascending infection.22 Fetotomy remains a delicate
the birth canal and the membranes have ruptured and use procedure in elephants and requires modified cattle equip-
of conservative measures has failed to progress parturition ment.22 So far, several fetotomy attempts have resulted in
should surgery be considered. Surgery is typically regarded death of the dam due to uterine trauma and infection.15
as the last resort to save the dam’s life. Cesarean section is, However, one successful description in an Asian elephant
however, not an option in elephants. To date, six cesarean exists.22 During this 9-hour procedure, all large fetal parts
sections have been carried out in elephants, all of which were extracted through the normal birth canal using ropes
resulted in the death of both mother and calf.15 This was passed through the vaginal vestibulotomy incision.22
due to the weight of the intestines and the large size of A common postsurgical complication that has been seen
the incision needed to extract an elephant calf, in com- following episiotomy is a persistent vestibular fistula.22 This
bination with mechanical forces and the slow healing of unnatural opening carries with it the risk of ascending
elephant skin. infection due to fecal contamination. Therefore trauma
of the tissue surrounding the incision should be avoided.
Episiotomy/Vaginal Vestibulotomy and Fetotomy Furthermore, second-intention healing of the skin while
Given the nature of the elephant reproductive tract’s only suturing the mucous membranes and the muscular
anatomy, with the vulval opening between the hind legs layer of the vaginal vestibulum are mandatory in order to
and the long canal of the vaginal vestibulum, no direct prevent this problem.
CHAPTER 96 Elephant Pregnancy and Parturition: Normal and Abnormal 687
19. Foerner JJ: Dystocia in the elephant. In Fowler ME, editor: Zoo 25. Doyle C, York B, Whitely A: A survey of Asian elephant births
and wild animal medicine: current therapy, ed 4, Philadelphia, PA, from 1962–1998, J Elephant Manag Assoc 10:146–148, 1999.
Saunders, 1999, pp 522–525. 26. Aroch I, Larian N, Avni-Magen N: Concentrations of free
20. Hartley M, Stanley C: Survey of reproduction and calf rearing in (ionized) and total calcium and magnesium in healthy captive
Asian and African elephants in Europe, JZAR 4:139–146, 2016. Asian elephants (Elephas maximus) and effects of sample type and
21. Mar KU, Lahdenpera M, Lummaa V: Causes and correlates of pH on measured free calcium and magnesium concentrations,
calf mortality in captive Asian elephants (Elephas maximus), PLoS Israel J Vet Med 71:24–32, 2016.
ONE 7(3):e32335, 2012. 27. Murray S, Bush M, Tell LA: Medical management of postpartum
22. Schaftenaar W: Delayed postpartum fetotomy in an Asian problems in an Asian elephant (Elephas maximus) cow and calf,
elephant (Elephas maximus), J Zoo Wildl Med 44:130–135, 2013. J Zoo Wildl Med 27:255–258, 1996.
23. van der Kolk JH, van Leeuwen JP, van den Belt AJ, et al: 28. Kowalski NL, Dale RH, Mazur CL: A survey of the management
Subclinical hypocalcaemia in captive Asian elephants (Elephas and development of captive African elephant (Loxodonta africana)
maximus), Vet Rec 162:475–479, 2008. calves: birth to three months of age, Zoo Biol 29:104–119, 2010.
24. Benfield RD, Newton ER, Tanner CJ, et al: Cortisol as a
biomarker of stress in term human labor physiological and
methodological issues, Biol Res Nurs 16:64–71, 2014.
97
Elephant Care in Southeast Asia
GERARDO MARTINEZ AND JOHN ROBERTS
689
690 SE C T I O N 19 Elephants and Rhinoceroses
Drugs for Field Immobilization needs to be “walked” (see explanation later), give 1 mg
White Rhinoceros atipamezole per mg medetomidine IV. This combina-
tion with oxygen supplementation at 10 L/min usually
In general, choose a dose that provides the lightest plane results in good blood oxygen levels. For a deeper level of
of anesthesia that is safe for completing the procedure. anesthesia, IV ketamine may be given.6
There are essentially two options for immobilizing white Newborn or very small calves may be adequately
rhinoceros (Ceratotherium simum). restrained with approximately 20 mg butorphanol IM. For
1. Potent Opioid + Sedative + Mixed Opioid Agonist- older calves, use 0.1 mg etorphine IM for every month of
Antagonist (Table 98.1) age up to 1 year, combined with IM butorphanol at 10–20
Intramuscular (IM) etorphine is combined with IM times the etorphine dosage in milligrams. When darting
azaperone or midazolam, and the respiratory depressant cows and calves, the cow is darted first and the calf 2–3
effects of etorphine1–3 are partially reversed with IM minutes later, or once the cow is recumbent.
or intravenous (IV) diprenorphine or butorphanol.4–9
This option is currently used for most white rhinoceros Black Rhinoceros
field immobilizations, particularly if animals are in good
condition and the terrain is challenging. Butorphanol is Etorphine is still the drug of choice at approximately 4 mg
routinely administered at 10–20 mg per mg etorphine IM for an adult male or female black rhinoceros (Diceros
IV once the rhinoceros is recumbent. This improves bicornis) in good condition.14 Adults of the subspecies D.
blood gas values and thereby indirectly reduces muscle bicornis bicornis may need 5 or even 6 mg of IM etor-
tremors.9–11 Tracheal or intranasal oxygen supplementa- phine.14,15 Younger animals are given a scaled-down dose
tion in combination with butorphanol may improve according to their size, and, for animals in poor condition,
hypoxemia.12 When butorphanol is administered in the be prepared to reduce the dose by 25% or more.
dart, the animal may stop but remain standing; this Thiafentanil is being used as an alternative in a 50:50
medication should be used only with experienced staff. combination with etorphine IM or by itself.14 The dose
Diprenorphine at 0.1–0.2 mg per mg etorphine IV8 may is the same as etorphine, induction is slightly faster, and
be given as an alternative to butorphanol after the animal the animals are less inclined to stay on their feet. Muscle
is recumbent. relaxation is generally good, but respiratory depression may
2. Mixed Opioid Agonist-Antagonist + Sedative (Table be more profound.
98.2) Approximately 60 mg of IM azaperone is usually added
Combining IM butorphanol with either medeto- to etorphine or thiafentanil for immobilizing an adult black
midine or azaperone6,13 is a good alternative when rhinoceros, although doses as high as 200 mg may be used.
immobilizing a compromised white rhinoceros. For Lower doses are preferred if the rhinoceros is to be trans-
adult white rhinoceros, 160 mg butorphanol + 10 mg located. IM midazolam is gaining favor as an alternative to
medetomidine gives standing sedation in 6–8 minutes azaperone at 20–40 mg for an adult (D. Grobler, personal
and recumbency at 12–15 minutes. The large volume of communication, 2017). Alternatively, 100 mg xylazine IM
drugs may be a limiting factor with this combination. for an adult is still being used.
Standing animals can be pulled down with ropes, or Hyaluronidase (2500–7500 IU) added to the immo-
200–400 mg ketamine can be administered IV to induce bilizing mixture reduces the induction time by a minute
recumbency. Reversal is achieved with administration or two, and this significantly reduces the risk in rugged
of IV naltrexone and atipamezole.6 If the rhinoceros terrain.14,15
692
CHAPTER 98 Updates in African Rhinoceros Field Immobilization and Translocation 693
TABLE Suggested Intramuscular Dosages When Using Etorphine and Butorphanol in Combination With a
98.1 Sedative in White Rhinoceros (Ceratotherium simum)
Midazolam (mg)
Age (10 × Etorphine Dose) Azaperone (mg) Detomidine (mg) Medetomidine (mg)
1 month — — — —
3 month 2 8 0.6 0.5
6 month 5 15 1.25 1
1 year 10 20 2.5 2
2 year 20 40 5 4
3 year 30 50 7 6
4 year 40 60 8 8
Adult 45 60 10 9
Large Adult 50 60 12 10
TABLE Suggested Intramuscular Dosages When Using Butorphanol in Combination With a Sedative in White
98.2 Rhinoceros (Ceratotherium simum)
*Use butorphanol and medetomidine, midazolam OR azaperone. Medetomidine is the sedative of choice because it may be easily reversed.
From Citino SB, Bush M: Reference cardiopulmonary physiologic parameters for standing, unrestrained white rhinoceros. J Zoo Wildl Med 38(3), 375–379, 2007.
Body Position and Blood Circulation to blade), anus, vulva, prepuce, nasal mucosa, eyelid, nictitat-
the Limbs ing membrane, buccal mucosa, and tongue.14
Using oxygen in the field is currently standard practice,
A rhinoceros in sternal position has better blood oxygen- and, when given intranasally, it rapidly improves blood
ation,10 but the lateral position supports improved circula- oxygen levels in both species. In white rhinoceros, it is
tion to the leg muscles, is better for cooling, gives access to most effective when given together with butorphanol and/
the udder, penis, and prepuce, and allows oral examination. or diprenorphine. It is of particular value in animals that
Whatever the position, make sure both nostrils are patent have undergone marked exertion and also in females that
and any fluid can freely run out of the mouth and nose. are heavily pregnant and in animals that are compromised
Depending on the circumstances, decide the best position or in poor condition. It is usually given at approximately
for the animal and be prepared to pump the animal’s legs 10–15 L/min, although higher flow rates may be initially
every 15 minutes to assist circulation, especially before indicated.
walking a rhinoceros into a crate.14
White Rhinoceros
Monitoring Breathing and Response to If breathing is unsatisfactory, partially reverse with 40 mg
Hypoxemia and Apnea butorphanol and/or 1.2 mg diprenorphine IV; this may be
repeated, but the animal may attempt to stand.
Respiratory depression is the biggest challenge and should
be the main focus of patient monitoring.10,12,16 A rate of Black Rhinoceros
6–12 breaths/min should be the aim, but slower breathing If ventilation is depressed, administer 5 mg butorphanol IV
(4–6 per minute) if deep and regular may be sufficient. immediately; this may be combined with 100–200 mg of
Oxygen saturation (SpO2) levels should be monitored. A IV doxapram. Some veterinarians give 2.5 mg butorphanol
pulse oximeter17 is an essential tool for rhinoceros immo- IV when they get to the rhinoceros and another 2.5 mg IM.
bilization; an SpO2 greater than 80% is usually adequate Butorphanol should be administered with caution because
for short procedures, with levels greater than 90% ideal. black rhinoceros may stand suddenly if given too much
The value of a pulse oximeter lies in observing trends, and butorphanol.
readings must be evaluated in conjunction with the rate For both species, if breathing stops, immediately inject
and depth of breathing, arterial blood color, heart rate, IV diprenorphine or naltrexone at calculated reversal dose,
and blood pressure. Clip-on or reflectance sensors can be put the rhinoceros in lateral recumbency, and apply pressure
applied to the ear (after both sides have been scraped with a to the abdomen, using your knee, to compress abdominal
CHAPTER 98 Updates in African Rhinoceros Field Immobilization and Translocation 695
Muscle Tremors
Muscle tremors are rarely a problem in black rhinoceros
but are frequently very marked in white rhinoceros,2 par-
ticularly if they are in the lateral position. Apart from being
caused indirectly by hypoxemia and acidosis,11 tremors also
increase oxygen consumption and heat generation,7 which
requires immediate intervention by giving butorphanol
and/or low-dose diprenorphine and oxygen.
• Figure 98.1 “Walking” a white rhinoceros (Ceratotherium simum).
Monitoring Heart Rate and Blood Pressure
Hypoxemia is associated with a sympathetic response that
increases the heart rate. Etorphine also causes hypertension per mg of etorphine, it may usually be stimulated to stand
and tachycardia in rhinoceros. Azaperone results in lower and walk by prodding; if unsuccessful, incremental doses
blood pressure when combined with etorphine.7 Butorpha- of diprenorphine (1 mg IV) may be administered. Alterna-
nol and/or low-dose diprenorphine administration lowers tively, if immobilized with etorphine and azaperone, give
heart rate, especially if combined with supplemental oxygen 10% of the standard diprenorphine reversal dose (1.2 mg
to counteract hypoxemia.3,7 for adults) IV immediately after the animal becomes
recumbent, to facilitate walking the rhinoceros (D. Cooper,
Hyperthermia personal communication, 2016). This approach results in a
slightly more alert animal, making walking easier, especially
To prevent hyperthermia, try to immobilize rhinoceros when uphill. If α2 agonists have been used for immobilization,
ambient temperatures are less than 25°C. Aim for a quick give incremental doses of atipamezole IV, starting at 1 mg
induction and minimize exertion. Once the rhinoceros is for every 1 mg medetomidine; keeping in mind that at 5 : 1,
recumbent, provide shade with a large beach umbrella and the effects of medetomidine will be fully reversed.
douse with copious cool water, using a handheld sprayer
and leaf blower for evaporative cooling of the animal. A Black Rhinoceros
thermoimaging camera is valuable to indicate the hotter
parts of the animal to focus cooling efforts. Never put a First roll the animal on its side for a minute or two and pump
wet rhinoceros in a closed crate in hot, humid conditions the legs; then put the rhinoceros into sternal recumbency,
without air flow, because heat stroke and shock may occur and administer 5 mg butorphanol IV. Wait 2–5 minutes for
quickly. the drug to have an effect and then stimulate the rhinoceros
The normal resting values for white rhinoceroses are by rocking it or using a prod. If the rhinoceros does not
given in Table 98.3.16 stand up, give an additional 5 mg butorphanol IV and
wait 2–5 minutes before stimulating again. Ten milligrams
“Walking” a Rhinoceros butorphanol is usually sufficient, but as much as 20 mg may
be needed. Black rhinoceros that have been immobilized
Rhinoceros must be securely blindfolded and have their with a 50:50 mix of etorphine and thiafentanil may also
ears plugged and a rope attached to the head and also to be walked with similar doses of IV butorphanol. As with
one of the hind legs to be used as a brake (Fig. 98.1). Eight white rhinoceros, keep the dose of azaperone low if you plan
people are typically needed to “walk” a rhinoceros, with two to walk a black rhinoceros or alternatively use midazolam.
(or three in the case of a white rhinoceros) supporting the Nalorphine is still used to walk black rhinoceros at
animal on either side and two on each of the ropes.14 10–20 mg IV instead of butorphanol, and some people are
comfortable walking black rhinoceros with a very low dose
White Rhinoceros of diprenorphine 0.25–0.4 mg IV.
blindfold and earplugs may be very effectively used to lip, because depth and rate of respiration are hard to see in
keep a rhinoceros calm, and a blindfold makes it easier to a moving vehicle. If dehydrated, administer polyionic fluids
quietly inject a rhinoceros in a crate. Adult black rhinoc- and dextrose and then use 1–2 mg of midazolam IM or IV
eros bulls are particularly challenging. Two techniques are to transport, repeating the dose if necessary. Ideally, plan
currently used: to have the midazolam wearing off (approximately 1 hour
1. Give 100–250 mg of the long-acting tranquilizer from last injection) on arrival at the orphanage. Attempt
zuclopenthixol IM and then walk into the crate with to bottle-feed the calf immediately on arrival, while in the
IV butorphanol given at 20 mg per mg of etorphine vehicle or crate, before reversing with IM or IV naltrexone
or thiafentanil used in the immobilizing dose. 1,2 mg and removing blindfold and earplugs. This technique may
diprenorphine is usually added to the butorphanol to save hours of struggling and works well for both black and
prevent pushing and head-pressing; this is sometimes white rhinoceros calves as young as 2–3 weeks (M. Toft,
necessary to repeat (IM or IV) after about 10 minutes personal communication, 2017).
if the animal continues to push. Additional tranquil-
ization on the road is usually 20 mg midazolam IM
or 60–100 mg azaperone IM, and later a low dose of Off-loading
etorphine (0.1–0.2 mg IM) might be necessary. Long White Rhinoceros
transports have been accomplished using this technique
with the rhinoceros sleeping much of the way. Some When off-loading or waking up a white rhinoceros in
people prefer to give as much as 60–80 mg butorphanol the field, use IV naltrexone at 20–30 times the etorphine
for an adult black rhinoceros, but head pressing may be dose in milligrams. Before reversal, remove all vehicles and
a problem and additional diprenorphine (0.3–0.6 mg IV personnel. In the case of a cow and calf, put the calf right
or IM) might be necessary to alleviate this. Additional next to the mother, facing the female. Reverse the calf first
tranquilization on the road is usually 20 mg midazolam and then the mother. Retreat quietly and monitor to ensure
IM or 20 mg butorphanol IM/IV or 60–100 mg azap- they do not get separated.
erone IM.
2. The alternative technique is to apply a cloth blindfold Black Rhinoceros
tightly to the head of the rhinoceros2 and plug the ears
with cotton wool, and then walk the rhinoceros into the They are much calmer at night; therefore arrange off-loading
crate, with 1.2–1.8 mg diprenorphine IV. The animal when it is dark, if feasible. Lights are unnecessary and usually
will be quite awake with this dose of diprenorphine, confuse the animal when used. A red headlamp works very
but because it cannot see, it will stand calmly in the well. If dark and the rhinoceros is heavily sedated, put the
crate. Midazolam or diazepam (10–15 mg IV) injected crate on the ground; then remove vehicles and administer
10 minutes before waking up may also improve the diprenorphine or naltrexone IM and immediately open
quality and duration of the sedation. If the rhinoceros the door.
struggles to stand in the crate, it may be stimulated During the day when a black rhinoceros is released from
by removing the earplugs or by prodding it on the a crate, it will inevitably attack the crate and any vehicles
forehead after waiting 60 seconds after drug injection. in the vicinity and risk seriously injuring itself. To prevent
An additional dose of diprenorphine (0.2–0.5 mg IV) this, one may either:
may be used if needed to reverse sedation. After approxi- 1. Inject the rhinoceros in the crate with an immobilizing
mately 4 hours, the effects of the immobilizing dose of dose of etorphine and azaperone. When it is about to
etorphine will start to wear off and 60 mg azaperone collapse, open the front door of the crate and hold the
and a very low dose of etorphine (0.05–0.1 mg) may be rhinoceros with ropes (either attached to the hind foot
administered IM every 2 hours as needed to maintain and/or put across its chest) until it falls. Alternatively, let
sedation. The rhinoceros should be closely monitored, the rhinoceros become recumbent in the crate and then
and if there is a lot of ear movement, indicating that it pull it out. If this option is used, a rubber mat on the
is lightly sedated, then more azaperone and etorphine is floor of the crate makes it much easier to pull out the
administered. A good response is generally seen within rhinoceros (J. Joubert, personal communication, 2017).
10 minutes. The crate, vehicles, and people must all be removed
from the area before the rhinoceros is reversed with
Transporting Orphan Calves IM/IV diprenorphine or naltrexone. Some people rub
the animal’s muzzle with its own dung and leave some
Calves are immobilized with a low dose of IM etorphine and of the dung close to the head before waking up (M.
azaperone depending on size, age, and condition, and then Toft, personal communication, 2017). This is also the
partially reversed with 1–2 mg butorphanol IV until stable. technique used for releasing a cow and calf combination
Place a blindfold (a bra works well) and plug the ears with both in the day and night. Place the immobilized calf
cotton wool and tape them closed. Monitor small calves close behind the cow and wake them up with naltrexone
during transport using a pulse oximeter on the tongue or or diprenorphine IM.
698 SE C T I O N 19 Elephants and Rhinoceroses
2. Alternatively, inject a low dose of etorphine (0.3–0.5 mg 9. van Zijll Langhout M, Caraguel CGB, Raath JP, et al: Evalu-
for an adult) IM and wait until the rhinoceros is heavily ation of etorphine and midazolam anesthesia, and the effect
affected; then inject a full dose of diprenorphine or of intravenous butorphanol on cardiopulmonary parameters in
game-ranched white rhinoceroses (Ceratotherium simum), J Zoo
naltrexone IM and immediately open the crate door.
Wildl Med 47(3):827–833, 2016.
The rhinoceros will wander off before the antidote 10. Wenger S, Boardman W, Buss P, et al: The cardiopulmonary
antagonizes the etorphine. effects of etorphine, azaperone, detomidine, and butorphanol in
field-anesthetized white rhinoceroses (Ceratotherium simum), J
References Zoo Wildl Med 38(3):380–387, 2007.
11. de Lange SS: Tremors in white rhinoceros (Ceratotherium simum)
1. Kock RA: Anaesthesia in zoo ungulates, J Assoc Vet Anaesth during chemical immobilization. Dissertation: University of
13:59–88, 1985. Pretoria, Faculty of Veterinary Science, 2015.
2. Radcliffe RW, Morkel PvdB: Rhinoceroses. In West G, Heard 12. Haw A, Hofmeyr M, Fuller A, et al: Butorphanol with oxygen
D, Caulkett N, editors: Zoo and wildlife immobilization and insufflation corrects etorphine induced hypoxaemia in chemically
anesthesia (vol 54). 2014, John Wiley & Sons, pp 741–771. immobilized white rhinoceros (Ceratotherium simum), BMC Vet
3. Hattingh J, Knox CM, Raath JP: Arterial blood pressure and Res 10(253):1–9, 2014.
blood gas composition of white rhinoceroses under etorphine 13. Radcliffe RW, Ferrell ST, Childs SE: Butorphanol and azaperone
anaesthesia, S Afr Tydskr Natuurnav 24:12–14, 1994. as a safe alternative for repeated chemical restraint in captive white
4. Boardman WSJ, Caraguel CGB, Raath JP, et al: Intravenous rhinoceros (Ceratotherium simum), J Zoo Wildl Med 31:196–200,
butorphanol improves cardiopulmonary parameters in game- 2000.
ranched white rhinoceros (Ceratotherium simum) immobilized 14. Morkel P, Kennedy-Benson A: Translocating black rhino: current
with etorphine and azaperone, J Wildl Dis 50:849–857, 2014. techniques for capture, transport, boma care, release and post-
5. Bush M, Citino SB, Grobler D: Improving cardiopulmonary release monitoring. IUCN SSC African rhinoceros Specialist
function for safer anesthesia of white rhinoceros (Ceratotherium Group 2007.
simum): use of opiate cocktails to influence receptor effects. 15. Morkel P: Chemical immobilization of the black rhinoceros
Proceedings AAZV, AAWV, AZA/NAG Joint Conference, 2005. (Diceros bicornis). Proceedings of a Symposium on “Rhino as
6. Bush M, Citino SB, Lance WR: The use of butorphanol in game ranch animals,” S Afr Vet Assoc 128–135, 1994.
anesthesia protocols for zoo and wild animals. In Fowler ME, 16. Citino SB, Bush M: Reference cardiopulmonary physiologic
Miller RE, editors: Fowler’s zoo and wild animal medicine: current parameters for standing, unrestrained white rhinoceroses, J Zoo
therapy, 2011, Elsevier, pp 596–603. Wildl Med 38(3):375–379, 2007.
7. Buss P, Miller M, Fuller A, et al: Cardiovascular effects of etor- 17. Fahlman Å: Advances in wildlife immobilization and anaesthesia;
phine, azaperone and butorphanol combinations in chemically clinical and physiological evaluation in selected species. Doctoral
immobilized captive white rhinoceros (Ceratotherium simum), J Thesis: Uppsala, Faculty of Veterinary Medicine and Animal
Zoo Wildl Med 47:834–843, 2016. Science, Department of Clinical Sciences, Swedish University of
8. Kock MD, Burroughs R, editors: Chemical and physical restraint Agricultural Sciences, 2008.
of wild animals. A training and field manual for African species,
IWVS(Africa) 223–233, 2012.
99
Update on Rhinoceros Nutrition
KATHLEEN E. SULLIVAN AND EDUARDO V. VALDES
H
uman greed and ignorance have led to devastation Although base nutrient recommendations in all extant
in dwindling wild populations; basic knowledge rhinoceros species have not been significantly altered in
of preventative health care is integral when rhi- recent history, research in the past 10–15 years has added
noceros are maintained in managed populations. Although specific knowledge based on species’ physiologic idiosyn-
nutrition under human care never replicates the variety crasies. Nutrient requirements have not been specifically
and exact composition of wild rhinoceros diets, informed detailed for each rhinoceros species; however, the horse is
feeding practices will serve to complement behavioral and considered the closest digestive model used for intake rec-
reproductive health. Several references exist on basic rhinoc- ommendations. Horse digestion and absorption have been
eros nutrition, including previous iterations of the current compared with rhinoceros species, with most applicability
text.1–3 Much nutrition research in the past 5–10 years nutritionally for white and greater one-horned rhinoceros,
has focused on the challenges of feeding rhinoceros under as compared with black or Sumatran.5 Rhinoceros have
human care, characterizing disease issues, and illustrating been recommended to be fed 1%–3% of their body
novel approaches. Researchers strive to make practical weight (BW) on an as-fed basis (1%–2% on a dry matter
connections between nutrient absorption, subsequent [DM] basis). It must be emphasized that a maximum of
physiologic and metabolic interplay, and repercussions on one-third of total calories comes from a pelleted concen-
reproduction and disease. Nutrient research in rhinoceros trate.1,3,6 Pelleted concentrate may vary in energy (calorie)
continues to be necessary as the complex interactions of content, especially compared with hay species, which
feedstuffs, genetics, and digestive physiologic needs of each possibly should be avoided while estimating that pellets
species continue to be discovered. themselves should be one-third of the diet. Monitoring
of energy intake is critical to maintaining healthy BW in
all species.
Generalized Recommendations for Good-quality forages (hay and browse) should provide
Feeding Rhinoceros Under Human Care primary nutrients for rhinoceros under human care, with
concentrate feeds used to balance energy, protein, mineral,
Rhinoceros consist of five extant species, the black (Diceros or vitamin needs.2,6 For browsing or intermediate species,
bicornis), white (Ceratotherium simum), Sumatran (Dicero- maximizing the amount of browse offered will be ideal;
rhinus sumatrensis), Indian or greater one-horned rhinoceros however, replicating natural browse variety consumed in
(Rhinoceros unicornis), and the Javan or lesser one-horned the wild is likely not practical or possible. All rhinoceros are
rhinoceros (Coelodonta antiqitatis). Rhinoceros are large adapted to use energy from fibrous plant material through
herbivorous mammals that currently have all species on the microbial fermentation. Animals should have ad libitum
International Union for Conservation of Nature Red List of access to hay, clean water, and salt.7 Feedings should be at
Threatened Species, with three species classified as critically least twice daily due to relatively fast transit time.1 Trace
endangered.4 They are primarily in danger from human mineralized salt would not be recommended, especially
poaching for their keratinous horn and habitat loss. Herbiv- for iron-sensitive species such as black and Sumatran
orous feeding strategies vary across species, with Sumatran, rhinoceros. Diets should be balanced through prioritizing
Javan, and black rhinoceros as browsers, white rhinoceros adequate roughage, limiting pelleted compound feeds, and
as grazers, and Indian rhinoceros classified as intermediate designing with mineral needs in mind for each species. It
feeders. Current worldwide population estimates (2017) should also be emphasized that not all pelleted feeds are
are 60–63 Javan, 100 Sumatran, 3500 Indian, 5042–5455 the same or interchangeable in terms of nutrients, and
black, and 19,682–21,077 white rhinoceros.4 Javan and use of cereal grain in pellets for all these species should
Sumatran rhinoceros are currently primarily maintained in be minimized. Abrasive pellets have been postulated to be
national parks in Indonesia, and this review will not focus connected to dental damage and tooth wear in rhinoceros
on nutrition of these species under human care. under managed care, unlike wild counterparts.8 However,
699
700 SE C T I O N 19 Elephants and Rhinoceroses
TABLE Mean Serum Mineral and Vitamin Values (±SD; N) for Black Rhinoceros Held Under Human Care
99.1 or Free Ranging
*Unpublished data from black rhinoceros under the care of Disney’s Animal Kingdom (1999–2016).
†
All vitamin values (see reference 12); all mineral values (see reference 11).
excessive starch and sugar content in some pellets and Best practices are recommended in the species-specific
produce may also be connected to this common issue sections later.
of severe dental plaque in rhinoceros under human care. For all species, regular monitoring of BW and condition
Black rhinoceros appear particularly prone to proliferative is also recommended because obesity under human care is
gingivitis, not always associated with degree of calculus well recognized as a risk factor and health detractor across
accumulation, also indicating dietary challenges.3 Therefore species. Use of body condition scoring (BCS) with a team
forage and browse, dependent on species, are integral to of animal caretakers with varied exposure to the animal
long-term health maintenance. It should be noted that (keeper, veterinarian, nutritionist, manager, etc.) will better
browse silage has been successfully fed to white and black maintain health goals during growth, pregnancy, or main-
rhinoceros species but may not be a practical production tenance. Communication across institutions to standardize
product due to labor and cost.9,10 BCS would also be beneficial due to its practical application
Although supplementation needs are limited in rhi- across disciplines.14
noceros under human care, attention should be paid to Rhinoceros milk, as described by Blakeslee and Zuba
mineral and vitamin balance in the diet for these species. (2002), describes a low level of total solids, high sugar,
For example, vitamin E in circulation appears lower low fat, and moderate protein that may be formulated for
across rhinoceros species compared with other mammals.1 hand-raising rhinoceros successfully using cows’ milk or
It should be ensured that total dietary vitamin E levels commercial milk replacer (Zoologic Milk Matrix 20/14,
reach the 150–200 IU/kg diet mark recommended by PetAg, Inc. Hampshire, IL).15 Analysis of milk across three
Dierenfeld for all rhinoceros species.1 Regular monitoring lactation period of a white rhinoceros found similar overall
of vitamin and mineral serum status in rhinoceros may nutrient values for fat and protein but altered fatty acid
serve to refine recommendations for feeding; however, composition compared with Milk Matrix 20/14, with higher
bioavailability across feed items for these nutrients is often levels of capric, lauric, and myristic acid.16 Guidelines for
variable. Reference values for animals both under human colostrum feeding of neonates and subsequent feeding rates
care and free ranging for minerals and vitamins may serve are clearly reviewed in the previous edition of this volume
as guidelines, but regular testing within institutions is and used with success at institutions across white, black,
critical for individual evaluations (Tables 99.1 and 99.2). and Indian rhinoceros species.3
CHAPTER 99 Update on Rhinoceros Nutrition 701
TABLE Mean Serum Mineral and Vitamin Values (±SD; N) for White Rhinoceros Held Under Human Care
99.2 or Free Ranging
*Unpublished data from black rhinoceros under the care of Disney’s Animal Kingdom (1999–2016).
†
All vitamin values (see reference 12); all mineral values (see reference 11).
TABLE
99.3 Example of White Rhinoceros Diet Design With Energy Contributions at Disney’s Animal Kingdom
*Mazuri Zulife Browser Rhinoceros Cube (5Z1P) Mazuri, Land O’Lakes, St. Louis, MO.
†
Stewart Products Inc. Bedford, TX.
‡
Omolene; Purina, Land O’Lakes, St. Louis, MO/DAK Mazuri Petting Zoo (5MJZ) Mazuri, Land O’Lakes, St. Louis, MO.
BW, Body weight.
TABLE
99.4 Example of Black Rhinoceros Diet Design With Energy Contributions at Disney’s Animal Kingdom
*Browse consists of three species in the winter (Elaeagnus and Acacia spp.) and three in summer (Willow, Banana, and Pennisetum spp.).
†
Mazuri ZuLife Browser Rhinoceros Cube (5Z1P); Mazuri, Land O’Lakes, St. Louis, MO.
‡
Stuart Products Inc. Bedford, TX.
§
Greens include bok choy, romaine, kale, endive, and green leaf Lettuce.
¶
Produce includes squash, celery, green beans, sweet potato, carrots, cauliflower, cucumber, zucchini, and bean paste.
**Other training items include pinecones, low-starch primate biscuit, and herbivore gel.
BW, Body weight.
substantial quantities of browse.29 Black rhinoceros diets both for concerns with obesity and metabolic syndrome, as
under human care may consist of pelleted feed, alfalfa and well as increasing bioavailability of iron. Caution and track-
grass-legume mixes, browse, and ideally a minority of enrich- ing should be exercised to limit training items, as well as
ment such as produce.28 Alfalfa hay should be limited, due high sugar and high iron items like molasses. Pellet formula-
to high protein, calcium, and iron in many harvests, as well tions for black rhinoceros are recommended to be low in
as creating diarrhea and colic.2,3,28 Black rhinoceros develop starch and soluble sugars, with NDF values from
iron overload disorder (IOD) under human care, where 40%–60%.28 An example of a successful diet for black
there is an excess of iron in circulation, measured repeatedly rhinoceros under human care that improved measures of
by iron biomarkers, and excessively stored, causing damage iron stores, primarily ferritin, is shown in Table 99.4.33
to tissues as demonstrated by necropsy.28,30,31 There also may Based on the practical limitations and availability of feed
be a recently found mechanism of iron absorption in play in items such as low-iron pelleted feed and browse, iron
black rhinoceros, which uses plant ferritin, a protein holding concentration in the diet was recommended to not exceed
up to 4500 atoms of iron, which exists only in legumes such 300 mg/kg DM or approximately 6 g of iron per day for
as alfalfa and absorbs it whole through clathrin-mediated a 1300 kg black rhinoceros fed 1.5% BW in DM.2,28 The
endocytosis.32 The possibility that legume iron stores may be commercial process of milling grains produces high-iron
absorbed outside of typically understood regulated absorp- pelleted commercial feeds, even in grain that is moderate
tion in the gut may be a key factor in contributing to in iron concentration.17,34 There have been positive correla-
IOD in this species with a need for further study. As previ- tions found between amount of grain-based products in
ously reviewed, excess generalized mineral supplementation the diets of European-held black rhinoceros and omega-6
should be avoided.5 Although a well-formulated pellet polyunsaturated fatty acids.35 Excess levels of omega-6 fatty
should provide adequate phosphorus and not overly high acids are established precursors to proinflammatory path-
calcium, extra supplementation as monosodium phosphate ways, which could exacerbate inflammation in an already
may be necessary, based on serum evaluations and for oxidant-sensitive species.
preventative reasons. Much work has focused on diseases of unknown etiology
Essential in formulating complete diets under human in the black rhinoceros, many of which may be tied to
care is limitation of high sugar and high vitamin C produce, inability to replicate wild nutrition for this species under
704 SE C T I O N 19 Elephants and Rhinoceroses
human care. This appears to include the interplay of but did not affect the apparent absorption of iron.46,47 Using
stress (both oxidant and environmental), improper fatty natural sources of tannins, such as grape pomace or tea
acid balance, lack of antioxidant, lack of polyphenolic leaves, there is doubt on the ability to supplement enough
antinutrients, excess of dietary iron, excess of starch and phenolic compounds to mimic levels observed in wild diets.6
other glucose supplies, and inadequate fiber in a diet under Grape seed extract was added as a form of natural chelator
human care.11,28,35 Antioxidant and phosphate supple- to both horse and black rhinoceros fecal continuous culture,
mentation appear to be preventative diet supplements for analyzing impact on the fecal microbial population.48 Grape
black rhinoceros.1 Physiologic findings indicate rhinoceros seed extract did not change nutrient digestibility or fer-
have a reduced capacity to neutralize oxidants, especially mentation, and similar microbial populations were found
black rhinoceros.36 Genetic testing of the black rhinoceros between species; the increased inclusion of the extract did
genome has identified specific mutations that may begin to increase condensed tannins and iron-binding capacity
explain the fragility of black rhinoceros red blood cells due and stimulated microbial growth.48 Although there were
to a mutation in their adenosine coding.37 no negative effects on the hindgut in vitro, more research
Theories exist on interconnection of obesity, insulin/ is warranted to understand in vivo effects, including pos-
glucose metabolism, metabolic dysregulation, and connec- sibly binding and limiting absorption of microminerals.
tion to iron loading and inflammation in black rhinoceros Recent work examined the use of an iron-specific drug
under human care.38 Baseline assays on banked samples chelator, known as HBED (N,N′-Bis(2-hydroxybenzyl)
from zoo-managed (n = 86) and free-ranging (n = 120) ethylenediamine-N,N′-diacetic acid), for oral use to
black rhinoceros were assessed for biomarkers of inflam- increase iron excretion in black rhinoceros under human
mation; with TNFα, serum amyloid A, insulin, insulin to care.42 The chelator was effective in increasing iron excre-
glucose ratio, and ferritin found statistically higher in zoo- tion through urinary output; however, due to one study,
managed versus free-ranging populations.38 This supports animal’s post study health crisis, administration of HBED
the need for investigation via longitudinal serial sampling is not recommended for black rhinoceros with documented
in zoo-managed black rhinoceros into factors impacting health problems.42 Iron-specific synthetic chelators may
inflammation, including nutrient and diet impact, as well still provide a future for preventative solutions, perhaps
as body condition and activity levels of these animals. Fluc- at lower or tapered doses.42 Although IOD is not the only
tuations of vitamin D with seasonal exposure to sunlight chronic disease state affecting black rhinoceros long term
(highest in summer) have been seen, despite vitamin D under human care, understanding the interplay between
supplementation in a pelleted feed year-round.39 Vitamin D, inflammatory pathways and disease progressions may lead
a known genetic regulator across species, also may fit into the to more concrete diet design for black rhinoceros. Future
complicated picture of metabolic regulation in this species. research must integrate practical husbandry concerns with
Comparisons between human iron overload and black epidemiologic perspective. Veterinarians, nutritionists, and
rhinoceros IOD indicate that black rhinoceros IOD husbandry professionals may then approach IOD preventa-
appears multifactorial, with both dietary iron intoxication tive measures with a similar mindset that emphasizes sharing
and intrinsic metabolic dysregulation as possible con- and centralizing information.
tributors.40,41 Although lowering dietary iron and increasing
natural dietary chelators to combat IOD in black rhinoceros Nutrition Moving Forward
may be a valid preventative approach to one aspect of this
disorder, classical human treatments of phlebotomy and Taking a preventative approach and trying to think long
iron-targeted synthetic chelation are equally valid options term in nutritional care serve to minimize nutritionally
with varied success.33,42 Measurements of iron biomarkers related disease states. Working across disciplines and think-
in free-ranging black rhinoceros (n = 194) found circulating ing of how nutrition, reproduction, behavior, and genetics
ferritin values, the only known marker of iron storage in are integrated in rhinoceros metabolism are necessary to
the liver, to be 290 ng/mL ± 18 ng/mL (mean ± SEM), move to sustainable populations. Practical diet guidelines
markedly lower than common under human care.43 A serve as a starting off point for balancing rhinoceros diets
black rhinoceros ferritin species-specific test is currently to maximize animal health.
under development based on a fully sequenced ferritin
gene; however, assessment of iron load should be made Acknowledgments
using ferritin and transferrin saturation (serum iron/
total iron-binding capacity).44 Thank you to Dr. Shana Lavin, Shannon Livingston, and
Chelators as a dietary therapy for IOD have been inves- Scott Williams for their aid in editing.
tigated. Condensed tannins (proanthocyanidins) are one
of a larger array of phenolic compound classes, which may References
work to decrease iron absorption in the digestive tract.45 It
has been demonstrated that inclusion at 0.5%–1.5% of diet 1. Dierenfeld ES: Rhinoceros feeding and nutrition. In Fowler ME,
DM as quebracho (a proanthocyanidin), but not tannic acid Miller RE, editors: Fowler’s zoo and wild animal medicine: current
(a hydrolysable tannin), increased total antioxidant capacity therapy, ed 4, Philadelphia, PA, 1999, Saunders, pp 568–571.
CHAPTER 99 Update on Rhinoceros Nutrition 705
2. Metrione L, Eyres A: Rhino husbandry manual, Fort Worth, TX, 21. Tubbs CW, Moley LA, Ivy JA, et al: Estrogenicity of captive
2014, International Rhino Foundation, p 327. southern white rhinoceros diets and their association with fertil-
3. Miller M, Buss P: Rhinoceridae (Rhinoceroses). In Miller RE, ity, Gen Comp Endocr 238:32–38, 2016.
Fowler ME, editors: Fowler’s zoo and wild animal medicine, ed 8, 22. Clauss M, Polster C, Kienzle E, et al: Studies on digestive
St. Louis, 2014, Elsevier, pp 538–547. physiology and feed digestibilities in captive Indian rhinoceros
4. International Rhino Foundation: State of the Rhino (www (Rhinoceros unicornis), J Anim Physiol Anim Nutr 89:229–237,
.rhinos.org). (Accessed 15 January 2017). 2005.
5. Clauss M, Castell JC, Kienzle E, et al: Mineral absorption in the 23. Clauss M, Polster C, Kienzle E, et al: Energy and mineral nutri-
black rhinoceroses (Diceros bicornis) as compared to the domestic tion and water intake in the captive Indian rhinoceros (Rhinoceros
horse, J Anim Physiol Anim Nutr 91:193–204, 2007. unicornis), Zoo Biol 24:1–4, 2005.
6. Clauss M, Hatt JM: The feeding of rhinoceros in captivity, Int 24. Huntington P, Pollitt C: Nutrition and the equine foot. Advances
Zoo Yearb 40:197–209, 2006. in Equine Nutrition III. 2005; 3:23.
7. Clauss M, Castell JC, Kienzle E, et al: Digestion coefficients 25. Sadler WC: The role of nutrition and its possible impact on
achieved by the black rhinoceros (Diceros bicornis), a large brows- elephant foot care. The elephant’s foot: prevention and care of foot
ing hindgut fermenter, J Anim Physiol Anim Nutr 90:325–334, conditions in captive Asian and African elephants, 2008, pp 13–15.
2006. 26. Bapodra P, Dierenfeld E, Wolfe BA: Evaluation of season-related
8. Taylor LA, Müller DW, Schwitzer C, et al: Tooth wear in captive dietary changes on the serum profiles of fat-soluble vitamins,
rhinoceroses (Diceros, Rhinoceros, Ceratotherium, Perissodactyla) mineral, fatty acids, and lipids in the captive greater one-horned
differs from that of free-ranging conspecifics, Contrib Zool rhinoceros (Rhinoceros unicornis), Zoo Biol 33:314–319, 2014.
83:2014. 27. Dutta DK, Bora PJ, Mahanta R, et al: Seasonal variations in food
9. Sullivan KE, Lavin S, Livingston S, et al: Palatability of bunker plant preferences of reintroduced rhinos Rhinoceros unicornis
ensiled willow as a winter diet item for browsing herbivores (Mammalia: Perrissodactyla: Rhinocerotidae) in Manas National
at Disney’s Animal Kingdom. Proceedings of the Nutrition Park, Assam, India, J Threatened Taxa 8:9525–9536, 2016.
Advisory Group to the Association of Zoos & Aquariums 9th 28. Clauss M, Dierenfeld E, Goff J, et al: IOD in rhinos – Nutrition
Biannual Conference, Kansas City, MO, 2011. Group Report: report from the nutrition working group of the
10. Hoellerl S, Stimm B, Hummel J, et al: Browse provision for international workshop on iron overload disorder in browsing
captive herbivores: design and management of a browse plana- rhinoceros (February 2011), J Zoo Wildl Med 43:108–113, 2012.
tion. Abstracts of the European Zoo Nutrition Meeting, 2005; 29. Helary SF, Shaw JA, Brown D, et al: Black rhinoceros (Diceros
4: 31. bicornis) natural diets: comparing iron levels across seasons and
11. Dierenfeld ES, Atkinson S, Craig AM, et al: Mineral concen- geographical locations, J Zoo Wildl Med 43:48–54, 2012.
trations in blood and liver tissue of captive and free-ranging 30. Molenaar FM, Sainsbury AW, Waters M, et al: High serum con-
rhinoceros species, Zoo Biol 24:51–72, 2005. centrations of iron, transferrin saturation and gamma glutamyl
12. Clauss M, Jessup DA, Norkus EB, et al: Fat soluble vitamins in transferase in captive black rhinoceroses (Diceros bicornis), Vet Rec
blood and tissues of free-ranging and captive rhinoceros, J Wildl 162:716–721, 2008.
Dis 38:402–413, 2002. 31. Olias P, Mundhenk L, Bothe M, et al: Iron overload syndrome
13. Flesness N: Normal hematological values, Apple Valley, CA, 2007, in the black rhinoceros (Diceros bicornis): microscopical lesions
International Species Information System. and comparison with other rhinoceros species, J Comp Pathol
14. Edwards KL, Shultz S, Pilgrim M, et al: Irregular ovarian activ- 147:542–549, 2012.
ity, body condition and behavioural differences are associated 32. Theil EC, Chen H, Miranda C, et al: Absorption of iron from
with reproductive success in female eastern black rhinoceros ferritin is independent of heme iron and ferrous salts in women
(Diceros bicornis michaeli), Gen Comp Endocrinol 214:186–194, and rat intestinal segments, J Nutr 142:478–483, 2012.
2015. 33. Mylniczenko ND, Sullivan KE, Corcoran ME, et al: Manage-
15. Blakeslee T, Zuba JR: Hand-rearing rhinoceroses. In Gage L, ment strategies of iron accumulation in a captive population
editor: Hand-rearing wild and domestic mammals, Ames, IA, of black rhinoceroses (Diceros bicornis minor), J Zoo Wildl Med
2002, Iowa State University Press, pp 236–244. 43:83–91, 2012.
16. Wertan N, Sullivan KE, Lavin SR, et al: Milk composition of 34. Koutsos E, Clauss M, Valdes E: Designing iron controlled diets
the southern white rhinoceros: A comparison across three preg- for exotic hoofstock – variability in raw materials and manufactur-
nancies in one animal. Proceedings of the Comparative Nutri- ing contributions to total dietary iron, Rio Grande, PR, 2016,
tion Society Symposium 9th Biannual Conference, Asilomar, Comparative Nutrition Society Symposium.
2012. 35. Clauss M, Dierenfeld ES, Bigley KE, et al: Fatty acid status in
17. National Research Council: Nutrient requirements of horses, ed 6, captive and free-ranging black rhinoceroses (Diceros bicornis),
Washington, DC, 2007, National Academies Press. J Anim Physiol Anim Nutr 92(3):231–241, 2008.
18. Shrader AM, Owen-Smith N, Ogutu JO: How a mega-grazer 36. Paglia DE, Tsu IH: Review of laboratory and necropsy evidence
copes with the dry season: food and nutrient intake rates by white for iron storage disease acquired by browser rhinoceroses, J Zoo
rhinoceros in the wild, Funct Ecol 20:376–384, 2006. Wildl Med 43:92–104, 2012.
19. Kiefer B, Gaslosser U, Kretzschmar P, et al: Food selection and 37. Paglia DE, Linzmeier R, Nemeth E, et al: Genetic basis of iron
food quality in territorial males of a free-ranging population of storage disease (ISD) in captive black rhinoceroses (Diceros bicor-
white rhinoceros (Ceratotherium simum simum) in South Africa, nis). Proceedings of the 15th Elephant and Rhino Conservation
Zoo Anim Nutr 2:199–207, 2003. and Research Symposium, Singapore Zoo, 2016.
20. Berkeley EV, Linklater WL, Dierenfeld ES: Dietary impact 38. Schook MW, Wildt DE, Raghanti MA, et al: Increased inflam-
on circulating glucose profiles in the white rhinoceros, J Anim mation and decreased insulin sensitivity indicate metabolic
Physiol Anim Nutr 95:245–251, 2011. disturbances in zoo-managed compared to free-ranging black
706 SE C T I O N 19 Elephants and Rhinoceroses
rhinoceros (Diceros bicornis), Gen Comp Endocr 217:10–19, bicornis) as a tool to understand factors affecting iron-overload
2015. disorder, J Zoo Wildl Med 47:820–826, 2016.
39. Oltman W, Olds J, Makowski AJ, et al: Seasonal variation of 44. Sullivan KE, Coffey R, Lavin SR, et al: Sequencing the black
25-hydroxy-vitamin D in two captive Eastern black rhinoceros rhino L-ferritin gene: how accurate is our testing? Proceedings
(Diceros bicornis michaeli). National Merial Veterinary Scholars of the Nutrition Advisory Group of the American Association of
Symposium, Ohio State University, 2016. Zoos & Aquariums Biannual Conference, Fresno, TX, 2017.
40. Klopfleisch R, Olias P: The pathology of comparative animal 45. Lavin SR: An overview of plant phenolics and their potential role
models of human haemochromatosis, J Comp Path 147:460–478, in mitigating iron storage disorder in wild animals, J Zoo Wildl
2012. Med 43:S73–S81, 2012.
41. Ganz T, Goff J, Klasing K, et al: IOD in rhinos – Immunity 46. Clauss M, Gehrke J, Hatt JM, et al: Tannin-binding salivary
Group Report: report from the immunity, genetics and toxicology proteins in three captive rhinoceros species, Comp Biochem
working group of the international workshop on iron overload Physiol A 140:67–72, 2005.
disorder in browsing rhinoceros (February 2011), J Zoo Wildl 47. Clauss M, Castell JC, Kienzle E, et al: The influence of tannin
Med 43:117–119, 2012. supplementation on digestive performance in captive black rhi-
42. Sullivan KE, Valdes EV, Livingston SE, et al: Use of a novel noceros (Diceros bicornis), J Anim Physiol Anim Nutr 91:449–458,
iron chelator (HBED) in black rhinoceros. In Bissell H, Brooks 2007.
M, editors: Proceedings of the 11th Conference on Zoo and 48. Huntley NF, Naumann HD, Kenny AL, et al: Black rhinoceros
Wildlife Nutrition, Portland, OR, 2015, Association of Zoos & (Diceros bicornis) and domestic horse (Equus caballus) hindgut
Aquariums Nutrition Advisory Group. microflora demonstrate similar fermentation responses to grape
43. Miller M, Chavey PS, Hofmeyr J, et al: Evaluation of serum seed extract supplementation in vitro, J Anim Physiol Anim Nutr:
ferritin and serum iron in free-ranging black rhinoceros (Diceros 1–15, 2016.
100
Health of the Forest Rhinoceros
of Southeast Asia: Sumatran
and Javan Rhinoceros
ROBIN W. RADCLIFFE AND KURNIA OKTAVIA KHAIRANI
707
TABLE Records of Disease Outbreaks With Associated Parasites and Pathogens in Captive and Wild Sumatran (Dicerorhinus sumatrensis) and Javan
100.1 Rhinoceros (Rhinoceros sondaicus)
(C) and (W) indicate captive and wild rhinoceros, respectively. References marked with an asterisk (*) are not peer reviewed.
Table prepared by Virgina Mule, DVM.
PCR, Polymerase chain reaction.
CHAPTER 100 Health of the Forest Rhinoceros of Southeast Asia: Sumatran and Javan Rhinoceros 709
TABLE Hematology and Biochemistry Values for Adult Captive Sumatran Rhinoceros (Dicerorhinus
100.2 sumatrensis)*
Index of
Analyte Mean ± SD Median IQR Min Max CVg (%) CVti (%) Individuality†
Hemoglobin (g/dL) 13.2 ± 1.1 — — 10.5 16.1 3 8 2.53
White blood cell count 7.1 ± 1.6 — — 3.2 12.2 17 16 0.89
(×103/µL)
Red blood cell count 5.1 ± 0.5 — — 4.1 6.5 4 8 2.33
(×106/µL)
Platelets (×103/µL) 133 ± 59 — — 18 280 24 40 1.68
Hematocrits (%) 39 ± 3 — — 32 48 2 8 3.53
Mean corpuscular 26 ± 1 — — 23 28 4 3 0.67
hemoglobin (pg)
Mean corpuscular 34 ± 1 — — 32 37 2 2 0.98
hemoglobin
concentration (g/dL)
Mean corpuscular 76.5 — — 62.7 94.1 — — —
volume (fl)
Protein (g/dL) 8.1 ± 1.1 — — 5.2 10.8 3 13 4.66
Albumin‡ (g/dL) – 3.9 3.6–4.5 1.9 7.3 1 17 24.70
Globulin (g/dL) 3.9 ± 1.3 — — 0.5 6.6 3 33 12.39
Aspartate 72 ± 23 — — 40 140 27 22 0.79
aminotransferase (U/L)
ALT‡ (U/L) – 21 16–31 7 69 5 14 3.05
LDH ‡,§
(U/L) – 884 684–1217 212 3583 <1 8 16.64
Bilirubin_Total (mg/dL) 0.6 ± 0.2 – – 0.3 1.2 14 31 2.25
§
Bilirubin_Direct (mg/dL) 0.3 ± 0.1 – – 0.01 0.6 2 52 23.68
Bilirubin_Indirect (mg/dL) 0.4 ± 0.2 – – 0.1 0.9 17 48 2.85
Serum urea (mg/dL) 21 ± 7 – – 6 42 20 31 1.55
‡
Creatinine (mg/dL) – 1.1 1.0–1.5 0.6 2.9 146 130 0.89
*Hematology and biochemistry values with estimates of central tendency (mean or median), variability (standard deviation [SD] or interquartile range [IQR]),
minimum and maximum reference interval values, between-animal coefficient of variation (CVg), within-animal coefficient of variation (CVti), and index of individuality
for adult captive Sumatran rhinoceros (Dicerorhinus sumatrensis) at the Sumatran Rhino Sanctuary, Lampung, Indonesia.
†
Estimated with the use of the equation CVti/CVg.
‡
The median and IQR are reported rather than the mean and SD because of the skewed distribution of these analytes; however, log-transformed variables were
used in mixed ANOVA models to calculate CVs and the index of individuality.
§
Ratu is excluded because of low sample numbers in order to estimate the between-individual animal variance.
Modified from Andriansyah, Candra D, Riyanto MA, et al. Hematology and serum biochemistry of Sumatran rhinoceros (Dicerorhinus sumatrensis) in a rainforest
sanctuary in Way Kambas National Park, Indonesia. J Zoo Wildl Med 44(2):280–284, 2013.
counts observed following a single trial. Because reinfection A 2003 outbreak in a captive population of Sumatran
was likely, control of flukes in a wet rainforest environ- rhinoceros housed in peninsular Malaysia at the Sungai
ment must also target the snail intermediate host through Dusun Conservation Center was attributed to infection
environmental interventions, such as clearing of vegetation with Trypanosoma evansi.18 The epidemic was characterized
around day stalls where rhinoceros feed. A Lymnea sp. snail by a biphasic die-off of animals with clinical signs that
was identified in a wallow frequented by the rhinoceros at varied from anorexia and depression to incoordination, rear
the sanctuary.10–12 limb paralysis, and recumbency. Pathology at the time of
the outbreak showed overgrowth of E. coli and Klebsiella
Ticks and Tick-borne Disease bacteria from multiple organ systems, generating significant
External parasites, including ticks, flies, and leeches, debate about the level of hygiene and husbandry at the
are common in the warm humid environments where sanctuary.19 However, subsequent histopathology revealed
Sumatran rhinoceros live. In addition to taking a blood that the bacteria were not associated with disease but rather
meal (which may offer a natural mechanism for iron consistent with overgrowth. Furthermore, trypanosomes
reduction), these parasites are vectors for important dis- invaded tissues and were found in various organs (includ-
eases. Of note are the hemoparasite infections carried by ing the brain), together with unique lesions in the spleen
ticks in the family Ixodidae. In a survey of four captive consisting of enlarged periarteriolar sheaths with lymphoid
Sumatran rhinoceros living in native rainforest habitat depletion, pathologic lesions characteristic of surra in other
in Way Kambas National Park, two species of ticks were mammals.16,18
identified, Haemaphysalis hystricis (81%) and Amblyomma
Noninfectious Disease
testudinarium (19%), with predilection for the neck and
shoulder skinfolds of the animals. At the time of the 2008 Eye Disorders
tick survey, simultaneous microscopic hematoparasite The Sumatran rhinoceros has a propensity for ocular dis-
examination of Giemsa-stained blood smears collected orders that is greater than that observed in other captive
from the captive rhinoceros revealed tick-borne diseases, rhinoceros species. Excessive exposure to ultraviolet light
including Anaplasma marginale (27%), A. centrale (10%), (UV) is the primary factor implicated in the ocular syn-
Babesia sp., and Theileria sp. (15%).10,13 Further molecular drome, although a multifactorial etiology is suspected given
analysis using reverse line blot hybridization (RLB) and the broad presentation of clinical signs in a variety of envi-
nested polymerase chain reaction (PCR) revealed Theileria ronments, including confinement within range countries.
bicornis in a single Sumatran rhinoceros. T. bicornis was first Eye conditions progress from mild corneal edema to severe
described as a novel blood parasite in free-ranging black opacity, uveitis, and blindness, with secondary bacterial and
rhinoceros (Diceros bicornis) in South Africa and, although fungal infections common sequelae. A case summary of a
fatal babesiosis from infection with Babesia bicornis was breeding pair of Sumatran rhinoceros in Sabah Malaysia
described in three black rhinoceros, there was no evidence compared development of clinical eye disease with indoor
that T. bicornis was associated with disease in African and outdoor locations in an attempt to elucidate causa-
rhinoceros.14 tion from light intensity or other environmental factors.
In an effort to boost immunity against tick-borne A seasonal pattern was noted, with all eye disorders
pathogens, one captive-born Sumatran rhinoceros destined appearing in the months of July and August, although no
for repatriation back to Indonesia from an American zoo correlation could be found to excessive UV exposure or dry
received three doses of a babesia-anaplasma vaccine (lyophi- conditions.20
lized protein of Babesia bigemina, B. bovis, and A. marginale The difference in light intensity in captive environments
of bovine origin) prior to the translocation.15 Although no compared with natural tropical forest architectures is likely
postvaccine titers were measured, the rhinoceros made a significant, given that chronic recurrent eye syndromes are
smooth transition into the tick-endemic environment of also prevalent in the captive Malayan tapir (Tapirus indicus)
Way Kambas, with mild subclinical hemoparasite infections coming from the same region. The rainforest environments
documented in serial blood smears. where these rhinoceros live consist of a complex forest struc-
ture in four layers, namely the emergent, canopy, understory,
Tabanids and Trypanosomes and forest floor. The extensive canopy and emergent layers
The emergence of animal trypanosomiasis (surra) in filter direct sunlight before it reaches the forest floor. In a
Sumatran rhinoceros highlights the growing threat of study of forest structure in Costa Rica, canopy architecture
pathogens transferred to novel hosts that have not adapted and light transmittance in both secondary and old growth
(or poorly adapted) to the agent.16 Trypanosomes evolved rainforests were compared—diffuse transmittance of light
on the African continent, and African rhinoceros have at 1–2 m above the forest floor (the understory level where
evolved a relatively stable host-parasite relationship, with Sumatran rhinoceros live) was less than 3%.21 Although
disease observed primarily during periods of stress or fol- shade structures in captive environments appear to help
lowing translocation of naïve animals into tsetse fly zones.17 reduce eye disease in this species, it is not always sufficient.
However, Asian rhinoceros are particularly susceptible and One captive Sumatran rhinoceros housed in an environ-
suffer high mortality. ment with extensive shade structures and high humidity
CHAPTER 100 Health of the Forest Rhinoceros of Southeast Asia: Sumatran and Javan Rhinoceros 711
developed eye disease during the winter months when the Smith et al. ferritin assay has been criticized by some
cloudy conditions predominated. because it requires species-specific reagents with variable
The integument of the black rhinoceros has been impli- cross-reactivity among diverse species and because results
cated as the primary organ in which allergic or disease sometimes seem highly variable in individual animals. The
conditions manifest under a variety of circumstances, sug- latter is likely due to serial dilutions of plasma that are
gesting that their epidermis is highly sensitive to disruption required for exceptionally high ferritin concentrations in
of metabolic homeostasis.22 The corneal epithelium of the species with captivity-induced ISD. In addition, results may
Sumatran rhinoceros eye may respond to disruptions in a be confounding because ferritin is an acute-phase reactant
similar manner with nutritional deficiency, UV exposure, that elevates secondarily in a number of inflammatory,
and dry conditions leading to increased disease states and neoplastic, or other conditions.
reduced healing—all of which are compounded with life Transferrin saturation, the amount of iron bound
in captive environments. The first sign of eye disease pre- to the plasma transport-protein transferrin, provides a
sents as corneal edema; then, if not treated with aggressive simple, qualitatively reliable, supplement or alternative
topical medication, peripheral vessel ingrowth occurs and if ferritin assays are equivocal or unavailable. Transferrin
pigmentation follows. Some animals progress to recurrent saturation correlates well with ferritin concentrations, with
anterior uveitis similar to moon blindness in horses (S. quantitative tissue analyses, and with histopathology using
Citino, personal communication, March 8, 2017). A vicious ferric-specific stains such as Prussian blue.24,27 Transferrin
inflammatory cycle of reinjury drives pathogenesis of ocular saturation in most vertebrates is approximately 35%. US
disease, with the initial insult causing inflammation that captive Sumatran rhinoceros measure 90%–100%, clearly
augments further injury to the ocular surface—invasion of indicating iron in sufficient excess to overwhelm carrying
leukocytes and release of immune mediators from damaged capacity of protective proteins.24,27 Alternative systems
cells leads to cyclic damage and reinjury. Therefore the for measuring ferritin and assessing ISD status have been
best response to treatment may be achieved using topical proposed, but these have not yet been validated by studies
cyclosporine ointment, an immunomodulator that inhibits directly comparing both assay systems or their relation to
T-lymphocyte proliferation (S. Citino, personal communi- demonstrable histopathology.28,29
cation, March 8, 2017). Nutrition is fundamental to the health of the browsing
rhinoceros, whether they are managed in captive or sem-
Iron Overload Disorder (Hemachomratosis, icaptive environments. A comparison of browse diversity
Iron Storage Disease) in Sumatran rhinoceros housed in three diverse settings
The induction of toxic overburdens of elemental iron in (North American zoo, Malaysian center, Indonesian sanc-
captive black rhinoceros was first noted by Smith et al. tuary) demonstrated marked differences in nutritional man-
(1995).23 Subsequent evidence extended these findings agement and predicted that these disparities relate directly to
to include Sumatran rhinoceros, the only other browser differences in iron loading.30 Browse diversity was measured
rhinoceros currently managed in captivity.24,27 Sumatran across five areas: number of locally available plant species,
rhinoceros develop progressive iron overloads even more number of plant species fed daily, access to a free-range
rapidly than do African black rhinoceros, reaching tenfold browse environment (i.e., native rainforest), transit time
elevations in body burdens within as little as 3 years of from plant cutting to feeding, and percentage of nonbrowse
captive birth or transfer into captive conditions and increas- items in diet (i.e., hay or pelleted ration). When comparing
ing in direct relation to time in captivity.24,27 traditional zoo rhinoceros with sanctuary animals, zoo rhi-
Measurements of serum ferritin concentrations and noceros were fed fewer species of browse (8 vs. 100 species);
transferrin saturation (the ratio of serum iron to total iron- fed fewer species on a daily basis (2–3 vs. 8–10 species); spent
binding capacity [TIBC]) provide the least invasive means fewer hours browsing (0 vs. 6 hours); ate browse that had
to assess iron status. It is widely acknowledged that serum been in transit longer (>72 vs. <12 hours); and fed signifi-
ferritin concentrations reflect total body iron stores with an cantly more nonbrowse items as percentage of diet (20%–
accuracy exceeded only by direct quantitative analyses of 38% vs. 0%). These same groups differed in iron stores, with
tissue samples.25 Most rhinoceros studies have relied on the zoo rhinoceros having higher mean ferritin than sanctuary
assay developed by Smith et al. (1984),26 which is available rhinoceros managed in range countries (2835 ± 295 ng/mL
through the Kansas State University Veterinary Diagnostic vs. 680 ± 168 ng/mL, respectively) (see also Chapter 99).
Laboratory. In separate studies, serum ferritin values mea- The inevitable morbidity and mortality of chronic
sured by this assay in African black and white rhinoceros progressive iron toxicity can be prevented by induction
(Ceratotherium simum) free-ranging in their natural habitats of negative iron balance through periodic phlebotomies,
were less than 100–200 ng/mL.27 By contrast, specimens as validated by extensive experience with an equivalent
from 14 captive Sumatran rhinoceros averaged greater human disorder, hereditary hemochromatosis. The clinical
than 850 ng/mL, with individual values ranging as high as effectiveness of this procedure has been validated in African
2000–4000 ng/mL. black rhinoceroses at multiple institutions, but has not yet
Despite its widespread use and verified correlation with been applied appropriately to Sumatran rhinoceroses (D.
quantitative tissue assays and necropsy histopathology, Paglia, personal communication, 2018).
712 SE C T I O N 19 Elephants and Rhinoceroses
Chronic Renal Disease (R. sondaicus sondaicus) solely inhabiting Ujung Kulon
A 30-year-old male Sumatran rhinoceros named Torgamba National Park (UKNP), with 67 individuals recorded in
housed at the Sumatran Rhino Sanctuary in Lampung, 2016; the Indian Javan rhinoceros (R. sondaicus inermis)
Indonesia developed renal disease characterized by pro- now extinct but once common throughout Bengal, Ban-
gressive azotemia, hypercalcemia, and hypophosphatemia gladesh, and Burma; and the Vietnamese Javan rhinoceros
(data summarized over a 4-year period showed progressive (R. sondaicus annamiticus) recently declared extinct in Cat
deterioration in renal function as measured by BUN 30.3 Tien National Park, Vietnam and formerly also found in
([10–52 µg/dL]; creatinine 3.2 [0.87–20.7 µg/dL]; Ca 15.8 Cambodia, Laos, Thailand, and Malaysia. Currently, many
[8.9–28.3 mg/dL]; Ph 3.3 [1.3–9.7 mg/dL]; and Ca to Ph challenges threaten the last population of Javan rhinoceros,
ratio 5.6:1 [1.2:1–11.8:1]. Radcliffe, RW and Candra D: including infectious disease at the wildlife–domestic animal
unpublished data, 2009). The disease was monitored with interface and noninfectious disease (toxic plants, invasive
serial measurement of body weight and serum biochemistry arenga palm, parasitism, and feeding competition with
analysis on a weekly basis. Nutritional management of the sympatric ungulates), all compounded by the significant
disease focused on feeding a highly palatable selection of demographic risk of natural disaster and inbreeding depres-
browse that included hand-feeding during periods of inap- sion inherent in a single small population. If not addressed,
petence together with supplementation of elemental phos- these challenges may create an irreversible extinction vortex.
phorus (Equi-phos; Uckele Health & Nutrition, Blissfield
MI 49228: guaranteed analysis of Ph 19%; Na 4.5%–5.5%, Javan Rhinoceros Mortality Events and
and Ca 0.1%–0.2%, with each ounce supplying 5.4 g of Infectious Disease
elemental Ph), electrolyte water, and a salt lick. Phosphorus
supplementation ranged from 1–4 oz per day, with dosing The first population census of UKNP was conducted in 1955
changes based on the most recent biochemistry panel; in by IUCN Director-General, Dr. Lee Talbot, and repeated a
general, the dose was increased by 1–2 oz per day when the dozen years later by WWF researcher, Professor Schenkel;
Ca to Ph ratio exceeded 5:1. The condition was managed both recorded fewer than 30 Javan rhinoceros.32,33 Since
successfully for half a decade before the rhinoceros finally then, the population has fluctuated between 58 and 69
deteriorated and died from complications of the disease. individuals. In 1982 the first of several mortality events was
reported, with five carcasses discovered with horns intact,
Reproductive Pathology and Allee Effect representing 8.9% of the population (see Table 100.1).33
The development of reproductive pathologies in female The investigation focused on infectious disease because the
Sumatran rhinoceros impacts both captive and wild con- park lies adjacent to local agricultural communities with a
servation programs. Uterine tumors, such as leiomyomas significant population of water buffalo (Bubalus bubalis).
and cystic endometrial hyperplasia, have been visualized The sequence leading to death was deduced from traces
on ultrasound and confirmed on histopathology.31 These at the site—walking and feeding, diarrhea, lying down,
diseases are more common in older animals and may be convulsive leg movements, and death. One comparatively
related to physiologic states related to long-term estrogen fresh carcass showed prolapse of rectum and foamy mucus
influence from cycling without pregnancy, a condition at the mouth and nostrils. Hemorrhagic septicemia (HS) is
observed in other captive rhinoceros. Uterine and ovarian an infectious disease caused by the gram-negative bacteria
masses have also been observed in recently captured wild Pasturella multocida. HS is a fatal disease of cattle, yak,
female rhinoceros, inferring that the same pathologies may camel, and water buffalo. In 1981 an HS outbreak was
be developing in wild animals. With their slow breed- responsible for the death of 350 domestic goats and 50
ing rate (long gestation and intercalving interval), small buffaloes in the region around the park. Anthrax outbreaks
populations of rhinoceros are particularly susceptible to were also recorded locally several decades previously. Despite
stochastic factors and the Allee effect (i.e., solitary nature inconclusive laboratory findings from the soil samples col-
and reduced mating opportunity, reproductive pathologies lected at the site, Schenkel concluded that anthrax was most
from prolonged periods of nonparity, skews in sex ratios, likely the causative agent because the spores are long lived
and inbreeding depression), with the end result being fewer and clinical signs were typical of acute outbreaks.
births than deaths.19 Following the 1982 die-off, the local government spon-
sored an HS vaccination program to districts that were
Javan Rhinoceros (R. sondaicus, affected by the outbreak, although the implementation
remains intermittent and irregular. For a 1-year period
Desmarest 1822) from June 2012 to July 2013, a disease surveillance study
Taxonomy was conducted to investigate the prevalence of HS.34 The
study was conducted in 19 buffer villages surrounding the
The Javan rhinoceros or one-horned lesser rhinoceros (R. national park due to a high risk of cross-infection between
sondaicus) is one of the most critically endangered terrestrial the villagers’ water buffalo and ungulates in the park,
mammals in the world. There are three distinct subspecies, including the Javan rhinoceros. Blood samples for serology
of which only one is extant; the Indonesian Javan rhinoceros (n = 770) and nasal swabs for culture (n = 85) were collected
CHAPTER 100 Health of the Forest Rhinoceros of Southeast Asia: Sumatran and Javan Rhinoceros 713
from water buffalo and compared with perceived risk factors size may impact fitness is significant for both species of
for buffalo herd management. A low seroprevalence of 1.8% Indonesian rhinoceros.19 Cooperative rhinoceros behaviors
(14 of 770 animals) was observed, suggesting that carrier such as feeding, breeding, territorial defense, and com-
animals could contribute to ongoing outbreaks in the park. munication through dung middens are less effective at low
Husbandry practices associated with a positive serologic population size, leading to decreased survivorship. Likewise,
response in water buffalo were: lack of a permanent area the per capita risk from predation and disease are height-
to house buffalo at night; low body condition score (BCS ened in small populations; a recent camera trap recording
= 2); high body temperature (fever ≥40°C); a history of documents predation of a banteng juvenile and a Javan
clinical signs or sudden death in the previous year; and a rhinoceros bull followed closely by a pack of Javan dhole
grazing system that accessed significant forage inside the (Cuonalpinus javanicus).
park. Serologic response was not associated with sex, age,
vaccination status, or season.34 Demographic Risks to a Single Population
Historic surveillance for endoparasites in Javan rhinoc-
eros fecal samples has demonstrated cestode, nematode, The loss of the Vietnamese subspecies of Javan rhinoceros in
and trematode infections with Strongyloides, Bunostomum, 2011 leaves UKNP as the last habitat for Javan rhinoceros
Trichostrongylus, Fasciola, Schistosoma, Anaplocephalidae, in the world. UKNP lies at the western most tip of Java
Oesophagostomoma, and Plagiotaenia.35–37 Protozoans Island in the heart of the Sunda Arc, an area of converg-
isolated include Balantidium, Entamoeba, Eimeria, Cripto- ing tectonic plates that commonly produces earthquakes
sporidium, Cycloposthium, Lavierella, and Ophryoscolecidae, and triggers tsunamis.41 In 1883 the eruption of Krakatoa
some of which are known pathogens in other species.37 devastated Ujung Kulon and its surrounding area, making
Vector-borne disease is emerging as a significant potential way for the Javan rhinoceros to colonize the region. Ironi-
threat to the UKNP Javan rhinoceros population, given the cally, the same threat that gave the Javan rhinoceros its last
disease reservoir of buffalo that are grazed inside the park. refuge is looming with Strombolian eruptions of Anak
Tabanid flies of the genus Tabanus are common hemo- Krakatau (Child of Krakatoa) actively spewing lava into
parasites known to transmit animal trypanosomiasis or the sea.42
surra. Trypanosomiasis infects cattle, water buffalo, horses, The creation of a second population of Javan rhinoceros,
elephants, camels, and rhinoceros, with Asian species being remote from Ujung Kulon in Cikepuh Wildlife Reserve, has
highly susceptible.16,18 Surveillance for T. evansi in 2014 been proposed to the government of Indonesia.43 Ecologic,
demonstrated a high prevalence (90%) of trypanosomiasis biological, and socioeconomic viability assessments are
in the livestock of two villages intersecting directly with underway to evaluate the readiness of Cikepuh to host a
the park boundary. A comprehensive study of tabanid founder population of four select Javan rhinoceros based
vector biology, including trypanosome infection rate and on distinct mean kinship. The second population strategy
host blood meal analysis, is underway to better understand is planned for execution in 2023.
host-parasite-vector dynamics in the UKNP ecosystem.38,39
Acknowledgments
Noninfectious Disease
The authors thank Dr. Donald Paglia for his contributions
One of the most challenging aspects of conserving the Javan to the Iron Storage Disease section and Dr. Scott Citino for
rhinoceros is the insufficient data regarding the species and insights on eye disorders. Dr. Virginia Mule collated Table
the habitat that shelters it. The Javan rhinoceros population 100.1 on literature reports describing pathogens, disease
has been on the rise since 1937; however, for the past 2 events, and mortality in Sumatran and Javan rhinoceros.
decades, population growth has been stagnant. A variety
of extrinsic and intrinsic factors may contribute to this
population plateau. References
Because vegetation analysis data in UKNP is limited, it
is possible that toxic plants have contributed to rhinoceros 1. Ashton KG, Tracy MC, de Queiroz A: Is Bergmann’s Rule valid
mortalities. An invasive palm (Arenga obtusifolia) known for mammals?, Am Natur 156(4):390–415, 2000.
locally as “langkap” may lower the carrying capacity for 2. van Strien NJ, Manullang B, Sectionov Isnan W, et al: Dicero-
browsers in the park. The invasive palm crowds out sunlight rhinus sumatrensis. The IUCN Red List of Threatened Species
and reduces secondary growth that provide the natural food 2008: e.T6553A12787457.
3. Furley CW: The clinical history of the adult female Sumatran
plants for the Javan rhinoceros. With the disappearance of
rhinoceros, called ‘Subur’, in captivity in Sumatra and at Port
open grasslands in the park, banteng (Bos javanicus) may be Lympne zoo, Kent, Great Britain. In Proceedings of Rhino,
competing with Javan rhinoceros for available browse. The Biology & Conservation, 1993, pp 357–366.
UKNP banteng population has been increasing from 200 4. Zainuddin ZZ, Rahman A, Omar S, et al: Severe necrotizing
in 1983 to more than 800 individuals in 2000.40 enteritis in a Sumatran rhinoceros (Dicerorhinus sumatrensis). In
Finally, the Allee effect may further limit Javan rhinoc- Proceedings of the First Congress of the Veterinary Association
eros population growth rates. The idea that population of Malaysia, 1989, pp 121–122.
714 SE C T I O N 19 Elephants and Rhinoceroses
5. Ludwig GV, Calle PP, Mangiagico JA, et al: An outbreak of West 22. Munson L, Miller RE: Skin diseases of black rhinoceroses.
Nile Virus in a New York City captive wildlife population, Am J In Fowler ME, Miller RE, editors: Zoo and wild animal
Trop Med Hyg 67(1):67–75, 2002. medicine: current therapy, ed 4, Philadelphia, PA, 1999, Saunders,
6. Abdelgawad A, Azab W, Damiani AM, et al: Zebra-borne equine pp 551–555.
herpesvirus type 1 (EHV-1) infection in non-African captive 23. Smith JE, Chavey PS, Miller RE: Iron metabolism in captive
mammals, Vet Microbiol 169:102–106, 2014. black (Diceros bicornis) and white (Ceratotherium simum) rhinoc-
7. Andriansyah, Candra D, Riyanto M, et al: Hematology and eroses, J Zoo Wildl Med 26:525–531, 1995.
serum biochemistry of Sumatran rhinoceroses (Dicerorhinus 24. Paglia DE, Dierenfeld ES, Tsu I-H: Pathological iron overloads
sumatrensis) in a rainforest sanctuary in Way Kambas National acquired in captivity by browsing (but not by naturally grazing)
Park, Indonesia, J Zoo Wildl Med 44(2):280–284, 2013. rhinoceroses. In Proceedings of the International Elephant and
8. Zainuddin Z-Z, Abdullah M-T, Shamsuddin M, et al: The Rhinoceros Resources Symposium, 2001, p 217.
husbandry and veterinary care of captive Sumatran rhinoceros 25. Lipschitz DA, Cook JD, Finch CA: A clinical evaluation of
at Zoo Melaka, Malaysia, Malay Nat J 1–19, 1990. ferritin as an index of iron stores, N Engl J Med 290:1213–1216,
9. Furley C: Veterinary work at Howletts and Port Lympne, Help 1974.
Newsletter 16–17, 1986. 26. Smith JE, Moore K, Cipriano JE, et al: Serum ferritin as a
10. Andriansyah DC, Muhammad A, Handayani SU, et al: Disease measure of stored iron in horses, J Nutr 114:677–691, 1984.
surveillance around Way Kambas National Park to support 27. Paglia DE, Tsu I-H: Review of laboratory and necropsy evidence
Sumatran rhino conservation and health. In Proceedings of for iron storage disease acquired by browser rhinoceroses, J Zoo
the Asian Zoo and Wildlife Medical Conservation, 2008, pp Wildl Med 43(3):S92–S104, 2012.
128–129. 28. Roth TL, Reinhart PR, Kroll JL: Development of a rhino ferritin
11. Schultz L, Radcliffe RW, Candra D, et al: Parasitology as a tool in specific assay for determining the association between serum
conservation medicine for health surveillance in captive and wild ferritin concentrations and hemochromatosis in the Sumatran
Sumatran rhinoceros (Dicerorhinus sumatrensis) with comparison rhinoceros (Dicerorhinus sumatrensis). In Proceedings of the
to sympatric populations, unpublished manuscript, 2009. 15th International Elephant and Rhinoceros Conservation and
12. Muryani A, Tiuria R, Andriansyah, et al: Helminthes parasite Resources Symposium, 2016, pp 68–69.
at feces of Sumatran rhinoceros (Dicerorhinus sumatrensis) 29. Watanabe M, Roth TL, Bauer SJ, et al: Feasibility study of NMR
and Sumatran elephant (Elephus maximus surnatranus) in Way based serum metabolomic profiling to animal health monitoring:
Kambas National Park Lampung (semi-in situ). In Proceedings a case study on iron storage disease in captive Sumatran rhi-
of the Asian Zoo and Wildlife Medical Conservation, 2008. noceros (Dicerorhinus sumatrensis), PLoS ONE 11(5):e0156318,
13. Sumantri CT, Hadi UK, Andriansyah, et al: The tick abundance 2016.
(Parastitiformes: Ixodidae) in Sumatran rhino in Way Kambas 30. Candra D, Radcliffe RW, Andriansyah, et al: Browse diversity
National Park Lampung and its role in diseases transmitting to and iron loading in captive Sumatran rhinoceroses (Dicerorhinus
Sumatran rhinceros (Dicerorhinus sumatrensis). In Proceedings of sumatrensis): a comparison of sanctuary and zoological popula-
the Asian Zoo and Wildlife Medical Conservation, 2008. tions, J Zoo Wildl Med 43(3):S65–S72, 2012.
14. Nijhof AM, Penzhorn BL, Lynen G, et al: Babesia bicornis sp. 31. Schaffer NE, Agil M, Bosi E: Utero-ovarian pathological complex
nov. and Theileria bicornis sp. nov.: tick-borne parasites associated of the Sumatran Rhinoceros (Dicerorhinus sumatrensis). In Pro-
with mortality in the black rhinoceros (Diceros bicornis), J Clin ceedings of the International Elephant and Rhinoceros Resources
Microbiol 41(5):2249–2254, 2003. Symposium, 2001, p 322.
15. Machado RZ, McElwain TF, Pancracio HP, et al: Babesia bige- 32. Schenkel R, Schenkel-Hullinger L: The Javan Rhinoceros (Rhi-
mina: Immunization with purified rhoptries induces protection noceros sondaicus Desm, 1822) in Ujung Kulon Nature Reserve,
against acute parasitemia, Exper Parasit 93:105–108, 1999. its ecology and behavior: field study 1967 and 1968, Acta Tropica
16. Desquesnes M, Holzmuller P, Lai D, et al: Trypanosoma evansi Separatun 26:2, 1969.
and surra: a review and perspectives on origin, history, distri- 33. Schenkel R, Schenkel L: Mystery of dead Javan rhinos, WWF/
bution, taxonomy, morphology, hosts, and pathogenic effects, IUCN Service 2(3):1982.
Biomed Res Int 2013, doi:10.1155/2013/194176. 34. Khairani KO, Nydam D, Felippe MJ, et al: Surveillance for
17. Emslie RH, Amin R, Kock R, editors: Guidelines for the in situ hemorrhagic septicemia in a buffalo (Bubalis bubalis) as an aid to
re-introduction and translocation of African and Asian Rhinoceros, range expansion of the Javan rhinoceros (Rhinoceros sondaicus) in
Gland, Switzerland, 2009, IUCN, p 115. Ujung Kulon National Park, Indonesia, J Wildl Dis 54(1):14–25,
18. Vellayan S, Mohamad A, Radcliffe RW, et al: Trypanosomiasis 2018.
(surra) in the captive Sumatran rhinoceros (Dicerorhinus suma- 35. Garrod AH: On the Taenia of the rhinoceros of the Sunderbunds
trensis sumatrensis) in peninsular Malaysia. In Proceedings of the (Plagiotaenia gigantea, Peters). In Proceedings of the Zoo Society
International Conference of the Association of the Institute of of London, 1877, pp 788–789.
Tropical Veterinary Medicine, 2004, pp 187–189. 36. Palmieri JR, Purnomo, Ammann H: Parasites of the lesser
19. Ahmad AH, Payne J, Zainuddin Z: Preventing the extinction of one-horned rhinoceros (Rhinoceros sondaicus desmarest), J Parasit
the Sumatran rhinoceros, J Indones Nat Hist 11(2):10–22, 2013. 66(6):1031, 1980.
20. Kretzschmar P, Sipangkui R, Schaffer N: Eye disorders in captive 37. Tiuria R, Primawidyawan A, Pangihutan J, et al: Identification
Sumatran rhinoceros (Dicerorhinus sumatrensis harissoni) in of endoparasites from faeces of Javan rhino (Rhinoceros sondaicus)
Sabah, Malaysia. In Proceedings of the International Conference in Ujung Kulon National Park, Indonesia. In Proceedings of the
on Diseases of Zoo and Wild Animals, 2009, pp 236–242. Asian Zoo and Wildlife Medical Conservation, 2006, pp 26–29.
21. Montgomery RA, Chazdon RL: Forest structure, canopy 38. Hariyadi ARS, Priambudi A, Setiawan R: Investigation of the
architecture, and light transmittance in tropical wet forests, Ecol death of Javan Rhinoceros (Rhinoceros sondaicus). In Ujung Kulon
82(10):2707–2718, 2001. National Park. Asian Soc Vet Pathol, 2011, pp 32–34.
CHAPTER 100 Health of the Forest Rhinoceros of Southeast Asia: Sumatran and Javan Rhinoceros 715
39. Radcliffe R, Khairani KO, Felippe J, et al: Tabanid fly host direction. Conservation Letter. Available at: http://onlinelibrary
feeding transmission patterns drive trypanosome infection in .wiley.com. (Accessed 12 April 2017).
endangered Javan rhinoceroses in Ujung Kulon National Park 42. Giachetti T, Paris R, Kelfoun K, et al: Tsunami hazard related to a
Indonesia, unpublished manuscript, 2017. flank collapse of Anak Krakatau volcano, Sunda Strait, Indonesia,
40. Yayasan Mitra Rhino. Laporan akhir: studi persaingan Badak Geol Soc London: Special Publ 361:79–90, 2012.
Jawa dan Banteng di Taman Nasional Ujung Kulon, unpublished 43. Setio P. Synthesis: survey result of Javan rhino’s (Rhinoceros son-
data, 2001. daicus, Desmarest 1822) future second habitat. WWF Indonesia.
41. Setiawan R, Gerber B, Rahmat UM, et al: Preventing global 2016.
extinction of the Javan rhino: tsunami risk and future conservation
Index
Page numbers followed by “f” indicate figures, “t” indicate tables, and “b” indicate boxes.
716
Index 717
Chlorhexidine Computed tomography (CT), in zoological medicine CPG. see Comprehensive Preparedness Guide
for minimally invasive surgery, 381 (Continued) C-reactive protein, for great ape cardiovascular
for reptiles and amphibians, 367 procedure planning in, 214 disease, 585
Chloroform, formalin, as treatment for scan settings in, 214–215 Crocodiles, herpesviruses in, 270
capripoxviruses, 645 review of, 212–214 Crocodilians
Chronic renal disease, in Sumatran rhinoceros, 712 data management in, 212 common clinical signs and differential diagnosis
Chronic wasting disease (CWD), 256t, 257–258 radiology expertise in, 213–214 of, 413t
clinical signs of, 257 for sea turtles, 346, 351f medical evaluation of, 412–418
control of, 259 Conciseness, of data, 24 age, 413
epidemiology of, 257 Cone beam computed tomography (CBCT), 210 body score, 413–414, 415f
transmission of, 255, 256f, 257–258, 258f Conenose bugs, as vector of Chagas disease, 241, circulatory system, 415
Chrysosporium anamorph of Nannizziopsis vriesii 241f digestive system, 416, 417f
(CANV), 394 Confounding bias, 22 history of, 412
Cidofovir, in papillomaviruses, 600 Consensus polymerase chain reaction (cPCR), in immune system, 414–415
Circoviridae, 269t virus detection and discovery, 112–113 integumentary system, 414
Circulatory system, of crocodilians, 415 Conservation, vaccines and, 299 laboratory examination, 417–418, 418f
CITES. see Convention on International Trade in Conservation biologists, in feral cats, 105–106 nervous system, 417
Endangered Species Conservation translocation, of potoroid marsupials, physical examination of, 413
Civets (Paguma larvata), SARS CoV and, 276–277 494–499, 495t reproductive system, 416
Clarithromycin, 407t conservation management, 494 respiratory system, 415–416
Clarity, of data, 24 health evaluation and disease surveillance in, 496 sex, 413
Clemacotyle australis, 331 postrelease monitoring after, 498 urinary system, 416
Climate change restraint, anesthesia, and analgesia in, 495–496, Cross-fostering, in macropod pediatric medicine,
anthropogenic, Saiga antelope and, 632–633 495f 505
effects on disease spread in wildlife, 247–254 telemetry, 496 Crossobothrium spp., in sharks, 341–342
distribution of pathogens, parasites, and transportation in, 496 Crossover trials, 59–60
vectors, 247–249, 248f welfare considerations in, 494–495 Cross-sectional studies, 61
emerging diseases, 250–251, 251f Consistency, of data, 24 Crustaceans, in saltwater public aquaria, treatment
prevalence or severity of disease, 249–250, 249f Contact premises, 49t protocols for, 326t–330t
recommendations on, 251–252, 252b Contamination, definition of, 145 Cryptic female choice, 131
Cloacal wash, for sharks, 340, 340f Contingency planning, 45 Cryptocaryon irritans, in saltwater public aquaria,
Clonidine, in chimpanzee, 575–576 principles of, 45–46 treatment protocols for, 326t–330t
Clostridial disease, 447b steps of Cryptomys damarensis, 514
Clostridium perfringens, in Saiga antelope, 632–633 determining goals and objectives, 47–48 Cryptosporidium hominis, in lemurs, 294
Clotrimazole, for birds, 444 formation of collaborative team, 46–47 Cryptosporidium spp., in orangutans, 566t–569t
Cohort studies, 60–61 plan development, 48, 49t Crystalloids, for elephant endotheliotropic
Cold stress, in giraffe, 619 plan preparation, review, and approval, 51 herpesvirus, 676t
Cold water hydrotherapy, for giraffe lameness, understanding the situation, 47 Culture, for elephant mycobacteriosis diagnosis, 665,
627t–628t Contraception 667t–668t
Committees, for zoo animal welfare, 68–70, 69t in African wild dog, 541, 542f, 542t Curators, in providing geriatric zoo animal end of
Common coquí (Eleutherodactylus coqui), 374–375 in captive Andean bear, 549 life care, 83, 85t
Common marmosets (Callithrix jacchus), MERS- in limiting reproduction, 131 Cutaneous invasive ascomycosis, in hibernating bats,
CoV and, 288 in surplus animals, 134–135 508–513
Communication Contrast enhancement, in computed tomography, clinical signs of, 509–510, 510f
during necropsy, 204 215 diagnosis of, 510–511, 510f
risk, 8–9 Control Access Point (CAP), 46t, 50 disease mitigation, 511
Complete blood count, for Zika virus infection, Control area biosecurity, 50 global perspective, 508
283 Control group, in crossover trials, 59–60 interdependence, host, and environment,
Complete crossover, in crossover trials, 59–60 Controlled mechanical ventilation (CMV), in 508–509
Compounding pharmacies (CP), 145–149 pinnipeds, 614 North American perspective, 508, 509f
in appropriate situations, 147–148 Controlled substances, in OHSP, 54 surveillance in, 511
definition of, 146–147 Convention on International Trade in Endangered Cutaneous respiration, in neonatal macropods,
developed from, 145 Species (CITES), 689 500
pharmaceutical legislative history and current of wild fauna and flora, 17–19, 19f Cuvier dwarf caimans (Paleosuchus palpebrosus),
perspective of, 145–146 labeling samples, 18–19 414
professional, 145 Nagoya Protocol, 18 CVD. see Cardiovascular disease
safety and efficacy parameters of, 145 packaging, 18–19 CWD. see Chronic wasting disease
traditional, 145 shipping and port of entry, 19 Cyclohexylamine plus alpha2-agonist, for free-
for zoo and wildlife specialty, relevance and veterinary import permits, 17–18 ranging lions, 537, 537t
considerations, 148 Cooperative State-Federal Brucellosis Eradication Cyclospora spp., in orangutans, 566t–569t
Comprehensive Preparedness Guide (CPG), 46t Program, 306 Cystic endometrial hyperplasia (CEH), in subfertility
Computed tomography (CT), in zoological Copepods, in saltwater public aquaria, treatment associated with regular cyclicity, 126
medicine, 206–217 protocols for, 326t–330t Cytokine stimulation assays, for elephant
advantages of, 222–223 Coral bleaching, 249, 249f mycobacteriosis diagnosis, 665–666, 667t–668t
benefits of, 206–208 Corn snake (Pantherophis [Elaphe] guttatus), 394, Cytology, sample collection, handling, and storage
bariatric, 208, 211f 396f in, 202t–203t
dental evaluation with, 207–208, 210f Coronaviridae, 269t Cytomegalovirus/chimp CMV, in orangutans,
superior detection, 206–207, 207f in marine mammals, 598 566t–569t
superior diagnostic capabilities, 206–207, 208f Coronavirus, 110–111, 115t
versatility, 207, 209f in bats, 276–277
virtual endoscopy, 208, 212f in Malagasy birds, 293–294 D
equipment considerations of, 208–210 Corticosterone, for stress evaluation, 66 DA2MP vaccine, in nondomestic carnivores,
access to, 208–209, 212f Corucia zebrata, 153 558t–560t
selection of, 209–210, 213f Cow, stem cell collection in, 139–140 Dactylogyroidea, in saltwater public aquaria,
interventional, 215, 216f Cownose rays treatment protocols for, 326t–330t
Mycobacterium pinnipedii and, 605 copper immersion in, 331–332 Damaraland mole rat, 514
optimization of, 214–215 Eimeria southwelli in, 331 Danofloxacin, 407t
artifact detection and prevention in, 215, 215f Coxsackie viruses, in orangutans, 566t–569t DCS. see Decompression sickness
contrast enhancement in, 215 cPCR. see Consensus polymerase chain reaction Deaths, of giraffe, 619
patient positioning and anesthesia in, 214 (cPCR) Decacotyle floridana, 331
Index 721
Decompression medicine Differentiation of Infected from Vaccinated Animals Ectotherms, euthanasia of (Continued)
in fish, 351–354 (DIVA) vaccines, 645 invertebrates, 357–360, 358t–359t
diagnosis of, 352, 352f Digeneans, in saltwater public aquaria, treatment reptiles, 361t
supportive medical treatment of, 354 protocols for, 326t–330t Edrophonium, in pinnipeds, 613
treatment of, 352–354 Digestive system, of crocodilians, 416, 417f Education programming, 29–30
in sea turtles, 345–350 Digital imaging, 13 EED. see Early embryonic loss
clinical history and symptoms of, 346 Digital Imaging and Communications in Medicine Egyptian fruit bat (Rousettus aegyptiacus), Marburg
diagnosis of, 346–348 (DICOM), 212 virus and, 276
diagnostic imaging of, 346, 350t format, 221, 224 Egyptian tomb bat (Taphozous perforatus), MERS-
laboratory profile of, 346 Digital radiology (DR), 218, 220–221 CoV and, 277, 288
postmortem studies of, 347–348 Dilaceration, 662 EHNV. see Epizootic hematopoietic necrosis virus
treatment of, 348–350 Dinoprostone, for elephants, 686 EHV-1 infection, 227, 228f, 229–230
Decompression sickness (DCS), 345 Dipeptidyl peptidase 4 (DPP4), 287 EHV-2 infection, 227
clinical history and symptoms of, 346 Diplotriaenoid spirurids, 477–478, 478f EHV-4 infection, 227
diagnosis of, 346–348 Direct methods, in monitoring arterial blood EHV-5 infection, 227
diagnostic imaging of, 346, 350t pressure, 187–188 EHV-7 infection, 227
laboratory profile of, 346 Disease mitigation, of white-nose syndrome, 511 EHV-9 infection, 227, 228f, 229–230
postmortem studies of, 347–348 Dispharynxosis, 471 EHVs. see Equine herpesviruses
treatment of, 348–350 DISS. see Diameter index safety system EIDs. see Emerging infectious diseases
Dehydration Distemper, in African wild dog, 542, 545t Eimeria southwelli
of confiscated turtles, 409 “Dive reflex,” in pinnipeds, 614 in Cownose rays, 331
in macropod pediatrics, 502t Diver’s disease. see Decompression sickness in saltwater public aquaria, treatment protocols
Delayed shedding, of molars, of elephant, 660–661 DMSO, for giraffe lameness, 627t–628t for, 326t–330t
Delivered anesthetic concentration, 178b DNA vaccine, in nondomestic carnivores, 561 Elasmobranch spp.
Delphinid herpesviruses, 599 DNA viruses, in marine mammals, 599–600 scuticociliatosis in, 332
Delta opioid peptide (δ or DOP), 151 Documentation, of training, 53–55 in touch-pools, 334–335
Demyelinating disease, in chimpanzee, in sanctuary, Dolphin (Delphinus delphis), brucellosis in, 310 ELDU. see Extralabel drug use
577–578 Domestic cats, 104–106, 105f Electrocardiogram (ECG)
Denaverine, for elephants, 686 adipose-derived mesenchymal stem cells, 140–141 in great ape cardiovascular disease, 585
Dengue virus, in orangutans, 566t–569t Domestic dogs, mesenchymal stem cells in, 140 for heart rate monitoring, 185
Dental disease, in elephants, 657–664 Domestic hosts, of Mycobacterium pinnipedii, 604b for heart rhythm monitoring, 186
diagnosis of, 659–660, 660f–661f Doppler imaging, in monitoring heart rate, 185 Electron microscopy, sample collection, handling,
management of, 657 Dosimetry bandages, 54–55 and storage in, 202t–203t
Dental evaluation, with computed tomography, Doxycycline, for giraffe lameness, 627t–628t Elephant care, in Southeast Asia, 689–691
207–208, 210f Dried alfalfa, in great ape nutrition, 590 captivity, 689
Dental formula, of captive Andean bear, 548 Drugs handling, 689
Dental pulp, of elephant tusk, 658–659 bulk, 145 history of, 689
Dependovirus, 115t generic, 145 Target Training Project and, 689–691, 690f–691f,
Depo-Provera injections, in limiting reproduction, pioneer, 145 691b
132 reactions, adverse, 145 Elephant endotheliotropic herpesvirus, 672–679
Depredation, cat, 106 use, extralabel, 145 clinical findings associated with, 674b–675b
Dermacentor albipictus, climate change on, 249 Dual-source CT, 210 endemic, 673t
Dermacentor spp., in capybaras, 521, 521f Dysbarism, definition of, 345 impact and epidemiology of, 672–673
Dermal fungal infections, in birds, 444 Dystocia preparedness, 673
Dermatophilus congolensis, in orangutans, 566t–569t in elephants, 684–685 treatment for, 674–675, 676t, 677f, 678b
Dermophthirius nigrellii, in sharks, 341 in subfertility associated with regular cyclicity, vigilance, 673–674
Desert tortoise, utility-scale solar PV plant on, 118 124 Elephant mycobacteriosis, 665–671
Deslorelin, for cystic endometrial hyperplasia, 126 diagnosis of, 665–666, 667t–668t
Deslorelin acetate (Suprelorin), for captive Andean occupational and public health considerations
bear, 549 E for, 670
Detomidine Eagles, utility-scale wind farm on, 118 transmission of, 665
for elephant endotheliotropic herpesvirus, 676t Early embryonic loss (EED), subfertility associated treatment of, 666, 670t
for white rhinoceros, 693t with, causes of, 128 Elephant plasma, for elephant endotheliotropic
in zoo and wildlife medicine, 166 Eastern bettong, 494 herpesvirus, 676t
Dewormers, for chelonian, 407t Eastern box turtles (Terrapene carolina carolina), 364 Elephants
Dexamethasone, for elephant endotheliotropic Eastern diamondback rattlesnake (Crotalus African, 658
herpesvirus, 676t adamanteus), 394 Asian, 658
Dexmedetomidine, 612 Eastern fox snakes (Pantherophis gloydi), 394–395 dental anatomy of, 657–659
in accidental veterinary anesthetic exposure, Ebola virus disease, 233–238 dental disease in, 657–664
171–173 in African apes, 233–234 diagnosis of, 659–660, 660f–661f
for fishes, 360t in Asian apes, 234 management of, 657, 658f
for reptile and amphibian, 424t background of, 233 treatment of, 662
in zoo and wildlife medicine, 166 in bats, 274, 276 dental morphology of, 658–659
2.5% Dextrose, 407t, 409 clinical signs of, 234 molar, 659, 659f
Diagnostic accuracy studies, 61 control measures for, 235–236 tusk, 658–659, 658f
Diameter index safety system (DISS), 182 diagnostics of, 235 diet of, 657
Diarrhea, in macropod pediatric conditions, differential diagnosis of, 235 eruption sequence of, 659
503t–504t future directions in, 236 normal parturition process in, 681–682, 682f
Diazepam, for chimpanzees, 575 reservoir for, 235 complications during, 684–685
Dicerorhinus sumatrensis. see Sumatran rhinoceros transmission of, 234–235 prediction of, 682–683, 683f–684f
Diceros bicornis. see Black rhinoceros Echinuria uncinata, 471 obstetrics in, 685–687
DICOM. see Digital Imaging and Communications Echocardiograms, in great ape cardiovascular disease, postpartum complications in, 687
in Medicine 584, 584f, 584t pregnancy in
Dicrocoeliidae spp., in orangutans, 566t–569t Ecologists, in feral cats, 105–106 abnormal, 683–684
Dientamoeba fragilis, in orangutans, 566t–569t Ectoparasites normal, 680–681, 681t
Dietary estrogenicity, of white rhino, 701 in Javan rhinoceros, 708t taxonomy of, 657
Diets in Sumatran rhinoceros, 708t ELISA. see Enzyme-linked immunosorbent assay
in great ape cardiovascular disease, 583–584 Ectotherms, euthanasia of, 357–363 Elk, brucellosis in, 307–308
great ape nutrition, 588, 589f amphibians, 357–359, 361t Embryonic stem cells (ESCs), 138
proportions of, 591, 591t–592t fishes, 360t Emerging infectious diseases (EIDs), 110, 111f, 113
722 Index
Emerging Pandemic Threats (EPT) program, 111 Estuarine crocodiles (Crocodylus porosus), 414 Female infertility (Continued)
Emydidae, herpesvirus infection in, 270 Ethambutol (ETH) associated with regular cyclicity, causes of,
Encephalitis for elephant mycobacteriosis, 666, 670t 124–126
in equine herpesviruses, 227 for Mycobacterium pinnipedii infection, 607 in zoo animals, 124–129
from Nipah virus (NiV), 275 Ethanol, for fishes, 360t Female reproductive status, monitoring of, 374, 376f
Encephalomyocarditis virus (EMCV), in orangutans, Etorphine Fenbendazole, 407t, 409
565–570, 566t–569t in accidental veterinary anesthetic exposure, Fentanyl, 152–153
End of life 169–171 in amphibians, 424t–426t, 426–427
of chimpanzee, in sanctuary, 578–579 for white rhinoceros, 693t in avian, 153
planning, for geriatric zoo animals, 83–91 in zoo and wildlife medicine, 164–165 in mammals, 152, 154t–158t, 160t
Endocrine disorders, in subfertility associated with Etorphine-xylazine, for musk ox, 637 in reptiles, 153, 424t–426t, 426–427
irregular cyclicity, 127 Etorphine-xylazine-midazolam-ketamine, for musk Feral cats
Endolimax nana, in orangutans, 566t–569t ox, 637 animal welfare in, 106
Endometriosis, in subfertility associated with regular Eugenol (clove oil), for fishes, 360t controversy in, 104–105
cyclicity, 126 Europe, babesiosis in, 647–651 stakeholders in, 105
Endometritis, in subfertility associated with regular European mink, vaccine-induced canine distemper dilemma, 104–109
cyclicity, 126 in, 556t public and wildlife health concerns related to,
Endoparasitism European reindeer (Rangifer tarandus tarandus), 106–107
in confiscated turtles, 409 Babesia spp. in, 648t–650t rise of, 104–105
in Sumatran rhinoceros, 709–710 Euthanasia. see also Trap-Neuter-Release (TNR) trap-neuter-release or euthanasia, management
Endoscopic biopsy of chimpanzee, in sanctuary, 578–579 alternatives to, 107
for African Clawed Frog (Xenopus laevis), 386f in macropod pediatric medicine, 505 veterinarians in, 107–108
for minimally invasive surgery in amphibians, Exhibits, for sharks, 338 Ferguson reflex, in elephants, 685
381–382 Exophiala pisciphila, in sharks, 341 Ferret badgers (Melogale spp.), SARS CoV and,
Endoscopic orchiectomy, of minimally invasive Exotic Newcastle disease, 451b 276–277
surgery in amphibians, 382–384, 385f Exotic ruminants, lumpy skin disease virus detected Ferritin, in Sumatran rhinoceros, 711
Endotheliotropic herpesvirus, elephant, 672–679 in, 643t Fertility, of white rhino, 701
clinical findings associated with, 674b–675b Exotic ungulate spongiform encephalopathy, 256t Fervac-D, in nondomestic carnivores, 558t–560t
impact and epidemiology of, 672–673 Experimental hosts, of Mycobacterium pinnipedii, Fetal retention, in elephants, 687
preparedness, 673 604b Fetotomy, for elephants, 686
treatment for, 674–675, 676t, 677f, 678b Exploratory behavior, in diagnosing behavior Fetus
vigilance, 673–674 problems, 77–78 malposition/ malformation of, causing dystocia, in
Endothermy, in neonatal macropods, 501 External examination, of necropsy, 198, 198b elephants, 684
Energy, and obesity in great ape, 593–594 clean-up and carcass disposal in, 200–201 oversized, causing dystocia, in elephants, 684
Eniconazole, for birds, 444 outbreak investigations in, 199–200 Fever, Zika virus infection and, 283
Enrichment critical components in, 200 Fibropapilloma lesions, of Green turtles (Chelonia
in giraffe, 620 species/age-specific considerations in, 199 mydas), 399f
in great ape nutrition, 592–593 Extracapsular cataract surgery, for cataracts, in flat, plaque-like cutaneous, 399f
Enrofloxacin (ENRO), 407t, 409 pinnipeds, 611–612 Fibropapillomatosis (FP), in marine turtles, 398–403
for elephant mycobacteriosis, 666, 670t Extralabel drug use (ELDU), 145 clinical signs of, 399–400
for giraffe lameness, 627t–628t Eye disorders, in Sumatran rhinoceros, 710–711 considerations of, 401
Entamoeba coli, in orangutans, 566t–569t Eye syndrome, of confiscated turtles, 409 diagnosis of, 400
Entamoeba hartmani, in orangutans, 566t–569t epidemiology of, 398–399
Entamoeba histolytica, in orangutans, 566t–569t etiology of, 398
Entamoeba spp., in orangutans, 566t–569t F prognostic indicators of, 401
Enterobius spp., in orangutans, 566t–569t Facial tumor disease, Tasmanian devil, 490–493 treatment of, 400
Enterovirus, 115t cause of, 490–491 Fibrous adhesions, in subfertility associated with
Environment, in behavior assessment, 77–78 devil populations, 491–492 regular cyclicity, 125
Environmental loggers, 12–13 moving forward, 492, 492f Field oxygen support, for free-ranging wildlife,
Environmental Protection Service (EPA), 4 signs and symptoms of, 490, 491f 181–182, 182f, 182t
Enzyme-linked immunosorbent assay (ELISA) FAD Preparedness and Response Plans (FADPReP), Field ventilatory support, for free-ranging wildlife,
on equine herpesviruses, 229 46t, 48 182–183, 183f
for Middle East respiratory syndrome, 289 FADs. see Foreign animal diseases Fifth sternebra, for stem cell collection, 139–140
for Zika virus infection, 284 Failure to thrive, in macropod pediatric conditions, Filarids, of birds, 477, 478f
EPA. see Environmental Protection Service 503t–504t File snake (Acrochordus spp.), 394
Episiotomy, for elephants, 686, 686f Falco sparverius, 160 Firocoxib, for giraffe lameness, 627t–628t
Epivax-TC-plus, in nondomestic carnivores, Fallow deer (Dama dama), and Babesia spp. in, Fisher exact test, 25–26
558t–560t 648t–650t Fishes, 357, 360f
Epizootic hematopoietic necrosis virus (EHNV), 367 False thumb, of captive Andean bear, 548 decompression medicine in, 351–354
Epizootic hemorrhagic disease (EHD) virus, climate Famciclovir diagnosis of, 352, 352f
change on, 248 for elephant endotheliotropic herpesvirus, 676t supportive medical treatment of, 354
Epsom salts, for giraffe lameness, 627t–628t for reptiles and amphibians, 367 treatment of, 352–354
Epstein-Barr virus/human herpesvirus 4, in Fan-beam CT, 210 euthanasia methods for, 360t
orangutans, 566t–569t Farmed mink, prion disease in, 256–257 necropsies on, 199
Equine herpesviruses (EHVs), 227–232 Fasciola spp., in Sumatran rhinoceros, 709–710 parasitic infections of, 323, 324b
background of, 227, 228f Feeding ecology, of captive Andean bear, 548–549 Flaviviridae, 268t–269t, 280
diagnostics of, 227–229 Feeding methods, in behavior assessment, 77–78 Flavivirus, 115t
experimental interspecies infections of, 229 Feline Leukemia Virus (FeLV), 107 Flavobacterium spp., in sharks, 341
modes of transmission of, 230–231 Feline spongiform encephalopathy (FSE), 256t Fluconazole, as antifungals in birds, 443
nonexperimental interspecies infections of, 230 incubation period for, 257 Flucytosine, 444
vaccination for, 231 Female anuran, hormones and dosages used, 377t Fluid therapy, for elephant endotheliotropic
Equine herpesvirus type 1 (EHV-1), in Sumatran Female frogs, exogenous hormone induction of herpesvirus, 676t
rhinoceros, 707 spawning in, 374–376 Flumazenil, for free-ranging lions, 537t
Ergonomics, OHSP and, 57 Female infertility, 124–128 Flunixin meglumine
Erythema, Zika virus infection and, 283 clinical history for, 125b for elephant endotheliotropic herpesvirus, 676t
Escherichia coli subfertility and for giraffe lameness, 627t–628t
in orangutans, 566t–569t associated with early embryonic loss, causes for reptile and amphibian, 424t, 429
sepsis, 449b of, 128 Fluoroquinolones, for elephant mycobacteriosis, 666
ESCs. see Embryonic stem cells associated with irregular cyclicity, causes of, FMD. see Foot-and-mouth disease
Estradiol, for elephants, 686 126–127 Follicle-stimulating hormone (FSH), 372
Index 723
Fomites, for equine herpesviruses transmission, 230 Gammaherpesvirinae, 227 Gonadotropin-releasing hormone (GnRH), 372,
Foot-and-mouth disease (FMD), 47 Ganciclovir, for elephant endotheliotropic 373f
annex using Secure Zoo Strategy, 48–51, 49t, 50f herpesvirus, 676t for African wild dog, 541
in Saiga antelope, 631–632 Garter snake (Thamnophis spp.), 394 Gongylonema, 471
Forages, for rhinos, 699–700, 702 Gas anesthesia, of musk ox, 637–638, 637f Good Samaritan law, 174
Foreign animal diseases (FADs), 46t Gas bubble disease (GBD) syndrome, 351–352 Gorillas
Foreign objects, in sharks, 342 Gas cylinders, 57 body weights for, 588
Forest rhinoceroses, of Southeast Asia, 707–715 Gas embolism (GE), 345 diets and digestive physiology of, 588, 589f
Formalin, degradation of, in a recirculating system, Gas exchange, in neonatal macropods, 500 Grass hay, for white rhino, 701
325 Gastric ghrelin secretion, in neonatal macropods, Grass-legume mixes, for black rhino, 702–703
Fossa (Cryptoprocta ferox), lyssavirus in, 294 500 Gray fox, vaccine-induced canine distemper in, 556t
Foxes, red (Vulpes vulpes), vaccinia-based General anesthesia, with analgesia, for minimally Great Ape Heart Project (GAHP), 581–587
recombinant vaccines targeting, rabies and, invasive surgery, 380–381 exam submission process, 583f
299, 300t–301t Generalist pathogens, 99 functions of, 581, 582f
Fractures Generic drugs, 145 Great ape nutrition, 588–595
in giraffe, 623, 624f–625f Genetic deep sequencing methods, for Middle East body weights in, 588
in tusks, of elephant, 657, 658f, 662 respiratory syndrome, 289 digestive physiology in, 588, 589f
Francisella tularensis, in orangutans, 566t–569t Genetic management, reproductive management feed presentation for, 592–593
Franquet’s epauletted fruit bat (Epomops franquetti), vs., 132 enrichment in, 592–593
Ebola virus in, 276 Genetic pool, in surplus animals, 135 group feeding in, 592
Free premises, 49t Genetics, of Trypanosoma cruzi, 241–242 health and, 593–594
Free-ranging lions, 536–538 Genitalia, abnormal, in subfertility associated with cardiovascular disease in, 594
capture technique for, 536 regular cyclicity, 124 obesity in, 593–594
immobilizing agents and antagonists, 536–538 Geopetitia, 471–476 orangutan air sacculitis in, 593
butorphanol, medetomidine, and midazolam, Geotrichum candidum, 395 recommended diet plan for, 588–592
537–538, 537t Geriatric zoo animals animal products in, 590
cyclohexylamine plus alpha2-agonist, 537, 537t end of life planning for, 83–91 browse in, 590
darting from helicopter, 538 quality of life assessment for, 83–91 diet proportions in, 591, 591t–592t
tiletamine-zolazepam (TZ), 536–537, 537t checklists for, 83, 86f–89f feeding guidelines in, 591–592, 592t
Free-ranging potoroids, diseases of, 498 flow chart for, 85–90, 90f fruit in, 590–591
Free-ranging timber rattlesnake (Crotalus horridus), models of, 83 kibble/pellets in, 589–590
396f Giant tortoise, in Galápagos Islands, 432–439 nuts in, 590
Free-ranging wildlife ecosystem restoration in, 436–437 pulses/legumes in, 590
anesthesia machines for, 179–181, 180f–181f, head-starting, 435–436 seeds in, 590
181b history of, 432–435, 433t, 434f–435f vegetables in, 590
basic inhalant pharmacology of, 177–178, 178b, restoration of, 435 vitamin and mineral supplementation in, 590
178t surrogate species and, 437–439, 438f water in, 588–589
field oxygen support for, 181–182, 182f, 182t taxonomy of, 432–434 whole grains in, 590
field ventilatory support for, 182–183, 183f Giardia spp., in orangutans, 566t–569t wild diets in, 588, 589f
gas anesthesia at altitude for, 179 Gilbert’s potoroo, conservation management for, Great apes
inhalant anesthesia for, 177–184 494 cardiovascular disease, 581–586
oxygen safety of, 182 Giraffe anesthesia considerations for, 585
patient monitoring in, 183–184, 183f anesthesia and, 623 biomarkers in, 585–586
vaporizers for, 177–184, 178b complications of, 620 blood pressure in, 582–584
Freshwater turtle (Pseudemys concinna concinna), arthropathies in, 623 cardiac health monitoring in, 584
herpesvirus infection in, 270 cold stress in, 619 cardiovascular changes in, 581–582
Frog virus 3 (FV3), 364 deaths of, 619 clinical signs of, 581
Fromm-D, in nondomestic carnivores, 558t–560t enrichment in, 620 diet in, 583–584
Fruit, in great ape nutrition, 590–591 exhibit design in, 621 echocardiograms in, 584, 584f
FSE. see Feline spongiform encephalopathy fractures in, 623, 624f–625f electrocardiogram in, 585
FSH. see Follicle-stimulating hormone hoof and limb disease in, 619–620 hypertension in, 582–584
Full-body alopecia, in captive Andean bear, 552, hoof overgrowth in, 623, 624f metabolic syndrome and, 584
552f husbandry and welfare in, 619–622 performing cardiac examinations in, 584, 584t
Full-term gestation, in animal breeding programs, keeper for, 621 postmortem cardiac evaluations in, 586
131 lameness in treating, 586
Fungal disease, in sharks, 341 clinical signs of, 626 experimental Ebola virus vaccine for, 303
Fungus Batrachochytrium dendrobatidis (Bd), 371 diagnosis of, 623–629, 626b impact of Ebola virus disease on, 233–234
Fur color, of captive Andean bear, 548 etiology of, 623 Greater one-horned/Indian rhino nutrition, 702
Fur seal (Callorhinus ursinus), brucellosis in, 312 management of, 623–629 Green anaconda (Eunectes murinus), 394
Furosemide, for elephant endotheliotropic operant training for, 628 Green power sources, in renewable energy, 117
herpesvirus, 676t prevention of, 628–629 Green tree pythons (Morelia viridis), Nidovirales
Fusarium solani, in sharks, 341 treatment of, 626–628, 626b, 627t–628t in, 271
laminitis in, 623 Green turtles (Chelonia mydas), 345, 398
nutritional disorders in, 619 fibropapilloma lesions of, 399f
G social structure of, 620 flat, plaque-like cutaneous fibropapilloma lesion,
Gabapentin, for giraffe lameness, 627t–628t training for, 621 399f
GAHP. see Great Ape Heart Project trauma of, 619–621, 620f severe multifocal fibropapillomatosis of, 399f
Galápagos Islands, giant tortoise relocation in, Global animal trade, infectious disease risk and, 4–5 Griseofulvin, for birds, 444
432–439 Global Virome Project, 277 Group feeding, in great ape nutrition, 592
ecosystem restoration in, 436–437 Glucocorticoids (GCs) GTAEF. see Golden Triangle Asian Elephant
head-starting, 435–436 assessment of, types of samples for, 73–74 Foundation
history of, 432–435, 433t, 434f–435f stress response and, 73, 74f Gyrodactyloidea, in saltwater public aquaria,
restoration of, 435 GNPD. see Galápagos National Park Directorate treatment protocols for, 326t–330t
surrogate species and, 437–439, 438f GnRH. see Gonadotropin-releasing hormone
Galápagos National Park Directorate (GNPD), 432 GnRH agonists, in limiting reproduction, 132
Galaxy 6-MPH-L, in nondomestic carnivores, Goals, in strategic planning, 35, 35b H
558t–560t Goatpox virus (GTPV), 641 HABs. see Harmful algal blooms
Galaxy-D, in nondomestic carnivores, 558t–560t Goiter, in sharks, 342–343 Haemaphysalis hystricis, in Sumatran rhinoceros, 710
Gamma interferon (IFN-γ) test, for elephant Golden Triangle Asian Elephant Foundation Haemoproteus, 293
mycobacteriosis diagnosis, 665–666, 667t–668t (GTAEF), 689 Haloperidol, in chimpanzee, 576
724 Index
Hammer-headed fruit bat (Hypsignathus monstrosus), Highly pathogenic avian influenza (HPAI), 263–264 Hypothermia
Ebola virus in, 276 H5N1, 264 for amphibians, 361t
Handling H5N2, 264 in giraffe, 619
of African wild dog, 541 H5N8, 264 in macropod pediatrics, 502t
of captive Andean bear, 549–550 High-throughput sequencing (HTS), in virus for reptiles, 361t
Hand-raising birds, 486 detection and discovery, 112–113 Hypoventilation, during anesthesia, 188
Hand-washing stations, in touch-pools, 336 Histology, sample collection, handling, and storage Hypovolemia, in macropod pediatrics, 502t
Hantavirus, 115t in, 202t–203t Hysterectomy
in orangutans, 566t–569t Historically controlled studies, 60 for captive Andean bear, 549
Harbor porpoises (Phocoena phocoena), brucellosis HMVCC. see Hungarian Meadow Viper for pyometra, 126
in, 310 Conservation Centre
Harbor seals (Phoca vitulina), brucellosis in, 310 Holtfreter’s solution, for amphibians, 386
Harmful algal blooms (HABs), 249–250 Homeless domestic cats, 104 I
Hawaiian monk seals (Neomonachus schauinslandi), Homing, of mesenchymal stem cells, 140 IBD. see Inclusion body disease
vaccination of, 299 Hoof cracks, in Sumatran rhinoceros, 708t IBDP. see Inclusion body disease protein
Hazard identification, 6 Hoof disease, in giraffe, 619–620 Ice slurry (rapid cooling), for fishes, 360t
HBOT. see Hyperbaric oxygen treatment Hoof overgrowth, in giraffes, 623, 624f Ichthyobodo spp., in saltwater public aquaria,
Health conditions, age-related, in zoo animals, 83, Horned vipers (Vipera ammodytes ammodytes), treatment protocols for, 326t–330t
84f–85f, 84t herpesvirus infection in, 270 ICS. see Incident Command System
Heart, Trypanosoma cruzi in, 242, 242f Horses IGRAs. see Interferon gamma release assays
Heifer syndrome, 307 Hendra virus in, 275 Iguana iguana, 153
Helicobacter, in orangutans, 566t–569t stem cell collection in, 139–140 IHA. see Indirect hemagglutination assay
Helicopter-based darting, of free-ranging lions, 538 Housing, of chimpanzee, in sanctuary, 575–576 Illegal activities, necropsy in, 194–195
Helminths HPAI. see Highly pathogenic avian influenza Imidocarb dipropionate, for babesiosis, 653
in Javan rhinoceros, 708t HTS. see High-throughput sequencing; High- Imiquimod, in papillomaviruses, 600
in Sumatran rhinoceros, 708t throughput sequencing (HTS) Immune assays, for marine mammal viruses, 600
Hemagglutination inhibition (HI) test, for Zika Human-based recompression protocols, in turtles, Immune system
virus infection, 284 349–350 of crocodilians, 414–415
Hematology Human HSV-1, in orangutans, 565 in neonatal macropods, 500–501
in babesiosis, 651 Human metapneumovirus, in orangutans, Immunofluorescence assay, for Burkholderia
of crocodilians, 414–415 566t–569t pseudomallei, 316
Hemochromatosis, in Sumatran rhinoceros, 708t Human Narcotic Safety Protocol, 54 Immunohistochemistry
Hemoparasites, in Sumatran rhinoceros, 708t Human predation, Galápagos giant tortoises, 434 of amphibians, 367
Hemorrhagic septicemia (HS), in Saiga antelope, Human T-lymphotropic virus-1, in orangutans, for marine mammal viruses, 600
631 566t–569t Immunology, in Mycobacterium pinnipedii infection,
Hendra viruses, bats and, 275 Humans 605–606
Henipaviruses, in bats, 275–277 atipamezole use in, 172f, 174 In situ hybridization (ISH) assay, for marine
Henry’s law, 345 disease risks from, 294 mammal viruses, 600
Hepadnaviridae, 269t naltrexone use in, 164–176, 171f Incident Command System (ICS), 46t, 47, 199–200
Hepadnavirus, 115t Human-wildlife interfaces, wildlife surveillance at, in Incisors, of elephants, 657–658
Hepatitis A virus (HAV), in orangutans, 566t–569t PREDICT approach, 112–113 Inclusion body disease (IBD), Arenaviridae and, 271
Hepatitis B virus (HBV), 113 Humoral immunity, in Mycobacterium pinnipedii Inclusion body disease protein (IBDP), 271
OHSP and, 55 infection, 606 Incubation, in animal breeding programs, 131
in orangutans, 566t–569t, 571 Hungarian meadow viper (Vipera ursinii rakosiensis), Indefinite hyposalinity, for Neobenedinia spp.,
Herbicides, embryonic death, in birds, 484–485 389f 324–325, 326t–330t
Herbivores, melioidosis in, 318 conservation action of, 388–389 Indian pythons (P. molurus), Nidovirales in, 271
Herpes B virus, 57 diagnostics of, 390–393, 391f–392f Indian rhino nutrition, 702
Herpes simplex viruses (1 and 2), in orangutans, life history traits of, 389t Indirect florescent antibody assay (IFA)
566t–569t management of, 389–390 in babesiosis, 652–653
Herpesvirales, 227 medical aspects of, 388–393 and Chagas disease in dogs, 243
Herpesviridae, 269–270, 269t medical considerations of, 389–390 Indirect hemagglutination assay (IHA), for
chelonian, 270 ovariohysterectomy in, 391f Burkholderia pseudomallei, 316
crocodilian, 270 results of, 388–389 Indirect immunofluorescence assays, for Middle East
in marine mammals, 599 special medical condition of, 390–393, 391f–392f respiratory syndrome, 289
squamate, 270 Hungarian Meadow Viper Conservation Centre Indirect (Riva-Rocci-”return of flow”) or noninvasive
Herpesvirus, 115t (HMVCC), 389 methods, in monitoring arterial blood pressure,
elephant endotheliotropic, 672–679 breeding results of, 390f 186–187
clinical findings associated with, 674b–675b Husbandry Indotestudo forsteni, 267
impact and epidemiology of, 672–673 of African wild dog, 539–540 Induced pluripotent stem cells (iPSCs), 138
preparedness, 673 of naked mole rats (NMRs), 514, 515f Infected premises, 49t
treatment for, 674–675, 676t, 677f, 678b regulatory roles and, 3 biosecurity for, 50
vigilance, 673–674 Hyaluronan, for giraffe lameness, 627t–628t Infection
equine, 227–232 Hydrochoerus hydrochaeris, 519 control, in OHSP, 55
background of, 227, 228f Hydrochoerus isthmius, 519 in fetotomy, in elephants, 686
diagnostics of, 227–229 Hydrochoerus spp. see Capybaras Infectious diseases
experimental interspecies infections of, 229 Hydromorphone, for reptile, 425t–426t in captive Andean bear, 553
modes of transmission of, 230–231 Hymenolepis spp., in orangutans, 566t–569t risk of, global animal trade and, 4–5
nonexperimental interspecies infections of, 230 Hyperbaric oxygen treatment (HBOT), 348 Infertility, in birds, 481
vaccination for, 231 Hyperbaric treatment, for fish, 352 Influenza, 115t
testudinid, 270 Hypercapnia Influenza A virus, in orangutans, 566t–569t
Herpetofauna, diversity of, in Madagascar, 292 during anesthesia, 188 Influenza B virus, in orangutans, 566t–569t
Heterocephalus glaber, 514–518 in pinnipeds, 613–614 Influenza viruses
biology of, 514 Hyperkeratosis, in Sumatran rhinoceros, 708t avian, 262–266
diseases in, 517, 517f Hypertension biosecurity of collections and vaccination of
husbandry of, 514, 515f in great ape cardiovascular disease, 582–584 nondomestic avian species against, 265
nutrition of, 514 during inhalation anesthesia, 187–188 changing ecology of, 262
reproduction of, 514–516, 516b, 516f Hypoglycemia, in macropod pediatrics, 502t classification of, 262
restraint and anesthesia for, 517 Hypotension, common under general anesthesia, highly pathogenic, 263–264
therapeutics for, 517 187 low-pathogenicity, 263–264
use in research, 517 Hypothalamic-pituitary-gonadal (HPG) axis, 373f paradigm of, 264–265
Index 725
Local anesthetics, for reptile and amphibian Male sterilization, in chimpanzee, 577 Melioidosis (Continued)
analgesia, 428 Male subfertility, in subfertility associated with etiology and epidemiology of, 315–316
Loggerhead sea turtle (Caretta caretta), 345 regular cyclicity, 124 factors predisposing to, 315–316
Logistic regression analysis, 25–26 Mammalian Meloxicam, 407t
Logistics, in mate acceptance, 131 buprenorphine in, 153–160, 154t–158t for giraffe lameness, 627t–628t
“Long-acting” (LA) formulation, definition of, 151 fentanyl in, 152, 154t–158t, 160t for musk ox, 640
Longevity, of zoo animals, 85 Mammals, necropsies on, 199 for potoroids, 495–496
Lophodont dentition, of elephant, 659, 659f Mammomonogamus laryngeus, in orangutans, Mentorship, 28–29
LOS. see Lines of Separation 566t–569t Meperidine, for reptile, 425t–426t
Low-pathogenicity avian influenza (LPAI), 263–264 Mammomonogamus spp., in orangutans, 566t–569t Meropenem, for melioidosis, 317
Low starch vegetables, based on animal weight, 592t Maned wolf, vaccine-induced canine distemper in, MERS. see Middle East respiratory syndrome
LRS or other balanced solution, 407t 556t Mesenchymal stem cells (MSCs), 138
Lumpy skin disease virus, detected in exotic Manual restraint, for sharks, 338–339 adipose tissue, 139–141
ruminants, 643t Marburg viruses, bats and, 274, 276 administration of, 139
Lupron Depot, for African wild dog, 541 Marine mammal viruses, 597–602 allogeneic, 142
Luteinizing hormone (LH), 372 diagnostics for, 600–601 bone marrow, 139–141
Lycaon pictus, 539–547 DNA viruses in, 599–600 homing of, 140
biology and anatomy of, 539 adenoviruses as, 599 in human and research animal medicine, 141
clinical pathology of, 541–542, 543t–544t herpesviruses as, 599 mechanism of function, 139
diseases of, 542–543, 544f–545f papillomaviruses as, 600 paracrine effects of, 141
distribution of, 540f poxviruses as, 600 regenerative potential of, 142
handling, restraint, and anesthesia for, 541, 543t RNA viruses in, 597–599 in veterinary medicine, 140
management, husbandry, and behavior of, astroviruses as, 597 xenogeneic, 142
539–540 Caliciviridae as, 598 in zoo medicine, 141–142
nutrition of, 540–541 Coronaviridae as, 598 Metabolic syndrome, in great ape cardiovascular
preventive medicine for, 544–545, 545t Orthomyxoviridae as, 598 disease, 584
reproduction and contraception for, 541, 542f, Paramyxoviridae as, 598–599 Metazoans, in saltwater public aquaria, treatment
542t viral biology in, clinical implications of, 597, 598t protocols for, 326t–330t
Lyme disease, climate change on, 248 Marine mammals, brucellosis in, 310–312, Methadone
Lyssavirus, in fossa (Cryptoprocta ferox), 294 310t–311t, 312f for amphibian, 424t
Marine turtles, fibropapillomatosis in, 398–403 for reptile, 425t–426t
clinical signs of, 399–400 Methicillin-resistant Staphylococcus aureus (MRSA),
M considerations of, 401 56
MAC. see Minimum anesthetic concentration diagnosis of, 400 Methocarbamol, for giraffe lameness, 627t–628t
Macaca fascicularis, 152 epidemiology of, 398–399 Method comparison studies, 61
Macaca mulatta, 152 etiology of, 398 Metronidazole, 407t
Macropod pediatric medicine, 500–506 prognostic indicators of, 401 Microbiology, sample collection, handling, and
approach to, 501–502 treatment of, 400 storage in, 202t–203t
anesthesia in, 502 Marsh deer (Blastocerus dichotomus), Babesia spp. in, Microneutralization (MN) assays, for Middle East
clinical pathology in, 502 648t–650t respiratory syndrome, 289
history in, 501, 501b Mass mortality events, in Saiga antelope (Saiga Microorganism amplifier, capybara as, 520–521,
pharmacology/toxicology in, 502 tatarica), 630–635, 631t, 632f 520f–521f
physical examination and assessment in, 501, discussion of, 632–633 Microsporum gypseum, in orangutans, 566t–569t
502t Mathematics of exposure, by zoo and wildlife Midazolam
common presentations, 502–505, 503t–504t veterinarians, 165t, 167t–168t, 169 for captive Andean bear, 550t
infectious diseases, 504 Matrix-assisted laser desorption/ionization time-of- for Hungarian Meadow Viper (Vipera ursinii
preventive medicine, 504–505 flight mass spectrometry (MALDI-TOF MS), rakosiensis), 391–392
cross-fostering in, 505 for Burkholderia pseudomallei, 316 for white rhinoceros, 693t
euthanasia in, 505 MBA. see Monterey Bay Aquarium Middle East respiratory syndrome (MERS),
normal postnatal development, 500–501 McNemar test, 25–26 287–291
Macropodidae, 500 Measles virus, in orangutans, 566t–569t bats and, 274, 276–277
Macropods, postnatal development of, 500–501 Medetomidine, 407t control and prevention of, 289–290
Macrorhabdus ornithogaster, 444 for accidental veterinary anesthetic exposure, diagnosis of, 289
Madagascar 171–173 epidemiology, 288–289, 288f
diversity of, 292 for captive Andean bear, 550t pathogenesis of, 287
endemic animals of, disease risk from introduced for chimpanzee, 576 public health response protocol for, 290
species to, 292–297 for white rhinoceros, 693t in reptiles, 271
from exotic carnivorans, 294–295, 294f for zoo and wildlife medicine, 166 transmission of, 289
from humans, 294 Media team, in providing geriatric zoo animal end treatment of, 289
from introduced amphibians, 293 of life care, 83–85, 85t virology of, 287
from introduced birds, 293–294 Medicine, sharks and, 338–344 Middle East respiratory syndrome coronavirus
from introduced rodents, 295 Medroxyprogesterone injections (Depo-Provera), for (MERS-CoV), from camels, 96–97
fauna of, 292 captive Andean bear, 549 Milk, from camels, 93–94
human colonization of, 292–293 Megestrol acetate (Ovaban), for cystic endometrial Mineral supplementation, in great ape nutrition, 590
Magnetic resonance imaging (MRI), in zoological hyperplasia, 126 Minimally invasive surgery (MIS)
medicine, 206–217 Méhely, Lajos, 388 of amphibians, 380–387
benefits of, 206–208 Melengestrol acetate applications of, fundamental research in,
equipment access to, 208–209 for captive Andean bear, 549 384–386
review of, 212–214 in limiting reproduction, 132 clinical applications of, 381–384, 381f–384f
data management in, 212 Melioidosis, 315–321 general considerations of, 380–381
radiology expertise in, 213–214 clinical presentation, pathology, treatment and postoperative management of, 386
of sea turtles, 346 management of, 316–319 recovery of, 386
Magnitude of the difference, 22–23 in birds, 318–319 definition of, 380
Mahouts, elephants and, 691, 691f in carnivores, 318 indications for, 381b
Mainstream analyzers, 189 in cetaceans, 316–317 Minimum anesthetic concentration (MAC), 178b
Makna, 658 in herbivores, 318 Mirounga angustirostris, 153
Male anuran species, hormones and dosages used to in pinnipeds, 317–318 MIS. see Minimally invasive surgery
induced spermiation in, 375t in primates, 318 Misoprostol, for elephants, 686
Male frogs, exogenous hormone induction of defined, 315 Missing values, 24
spermiation in, 372–374, 374f diagnosis of, 316 Mission, in strategic planning, 34, 35b, 36
Index 727
MLST. see Multilocus sequence typing Mycobacterium tuberculosis infection, in elephants, Nociceptin opioid peptide (NOP), 151
Modified-live virus vaccines, in nondomestic 665–671 Nociceptin receptor, 151
carnivores, 558t–560t diagnosis of, 665–666, 667t–668t Nonanesthetized great apes, echocardiograms on,
Molars, of elephant, 658–659, 659f occupational and public health considerations 584
diseases/anomalies affecting, 660–661 for, 670 Nonclinical painted turtles (Chrysemys picta), 368
treatment of, 662 transmission of, 665 Nondomestic avian species, vaccination of, against
supernumerary, 661 treatment of, 666, 670t avian influenza viruses, 265
Molecular assays, for Middle East respiratory Myiasis, in Sumatran rhinoceros, 708t Nondomestic carnivores, canine distemper
syndrome, 289 vaccination in, 555–563, 556t
Molecular diagnostics, sample collection, handling, modified-live virus vaccines, 558t–560t
and storage in, 202t–203t N recombinant viral vector vaccines, 555–557,
Monitored premises, 49t 0.45% NaCl, 407t, 409 558t–560t
Monkeypox virus, in orangutans, 566t–569t Nagoya Protocol, 18 Nonhuman primate (NHP)
Monoclonal antibody, for Burkholderia pseudomallei, NAHEMS. see National Animal Health Emergency disease prevention from, 57
316 Management System global pandemic infections in, 110
Monogeneans Naked mole rats (NMRs), 514–518 prion-infected, 257
in saltwater public aquaria, treatment protocols biology of, 514 transmissible spongiform encephalopathies in,
for, 326t–330t diseases in, 517, 517f 256t
in Spotted eagle Rays, 331 husbandry of, 514, 515f Nonmammalian taxa, lifetime reproductive planning
Mononegavirales, 115t nutrition of, 514 in, 133
Monterey Bay Aquarium (MBA), 354 reproduction of, 514–516, 516b, 516f Nonrandomized studies, 60
Moose (Alces alces) restraint and anesthesia for, 517 Nonsteroidal anti-inflammatory drugs (NSAIDs)
Babesia spp. and, 648t–650t therapeutics for, 517 for giraffe lameness, 627t–628t
brucellosis in, 308 use in research, 517 for musk ox, 640
Morbillivirus, in marine mammals, 598–599 Nalmefene, in zoo and wildlife medicine, 165–166 for reptile and amphibian analgesia, 428–429
Morphine Naloxone NOP. see Nociceptin opioid peptide
for amphibian, 424t for amphibian, 424t North America, babesiosis in, 647–651
for reptile, 425t–426t for reptile, 425t–426t Northern map turtle (Graptemys geographica),
Mpala Research Center (MRC), 94 for zoo and wildlife medicine, 165–166 herpesvirus infection in, 270
MRC. see Mpala Research Center Naltrexone Novel viruses, 113, 115t
MRSA. see Methicillin-resistant Staphylococcus aureus for elephant endotheliotropic herpesvirus, 676t NRC-NAS. see National Research Council of the
MSCs. see Mesenchymal stem cells in emergency response to human exposure, National Academy of Sciences
MTBC. see Mycobacterium tuberculosis complex 164–176, 171f NSAIDs. see Nonsteroidal anti-inflammatory drugs
Mu opioid peptid (µ or MOP), 151 for free-ranging lions, 537t Nucleic acid-based techniques, for marine mammal
Mucor circinelloides, in sharks, 341 National Animal Health Emergency Management viruses, 600–601
Mucosal fungal infections, in birds, 444 System (NAHEMS), 46t Numida meleagris, 153
Mule deer (Odocoileus hemionus), Babesia.spp. in, National Research Council of the National Academy Nutrigel (Virbac), for amphibians, 386
648t–650t of Sciences (NRC-NAS), 4 Nutrition
Multidetector CT, 210 Native wildlife, disease risks to, 99–103 of African wild dog, 540–541
Multi-host pathogens, 99 pathogens in, 99–100, 100t in captive Andean bear, 548–549
Multilocus sequence typing (MLST), of Burkholderia risk analysis for, 101–103, 102t great ape, 588–595
pseudomallei, 315 Nautilus pompilius, 196 of naked mole rats (NMRs), 514
Multimodal analgesic approaches, in reptile and NDOW. see Nevada Department of Wildlife rhino, 699–706
amphibian, 424t–426t, 429 (NDOW) black rhino, 702–704, 703t
Multivariable analyses, 23f, 26b NDV. see Newcastle disease virus greater one-horned/Indian rhino, 702
Mumps virus, in orangutans, 566t–569t Necropsy, 57 moving forward, 704
Musk ox (Ovibos moschatus), 636 internal examination of, 198, 198b recommendations for feeding, 699–700,
sedation and anesthesia, 636–640 clean-up and carcass disposal in, 200–201 700t–701t
biology of, 636 outbreak investigations in, 199–200 white rhino, 701, 702t
distribution, habitat, size, and weight, 636 species/age-specific considerations in, 199 in subfertility associated with irregular cyclicity,
environmental considerations of, 636 Necrotizing enteritis, in Sumatran rhinoceros, 708t 127
immobilization of, 637–638 Needle stick exposures, by zoo and wildlife Nutritional bone disease, of crocodilians, 416
monitoring during, 639–640 veterinarians, 167t–168t, 169 Nutritional disorders, in giraffe, 619
perianesthesia, 638–640, 639f Nematodes, in saltwater public aquaria, treatment Nutritional support, for sharks, 343
physical and chemical restraint of, 636–637, protocols for, 326t–330t Nuts, in great ape nutrition, 590
637f Neonatology, in birds, 486–487, 487b
recommended doses for, 638t Neoplasia
restraint/sedation/immobilization of, 636–640 in African wild dog, 543 O
use of long acting tranquilizers, 638 in captive Andean bear, 553 Obesity
Mycobacterium avium, in orangutans, 566t–569t Neoplastic ovarian tumors, 127 in great ape, 593–594
Mycobacterium bovis, long infectious period of, Neotropical primates, Zika virus and, 282 in zoo animals, 127
99–100 Nervous system, of crocodilians, 417 Observational studies, 21–22
Mycobacterium pinnipedii infection, 603–609 Neuromuscular blockage, in pinnipeds, 613 Obstetrics, in elephants, 685–687
antemortem diagnosis of, 605–606 Nevada Department of Wildlife (NDOW), 118 Occupational Health and Safety Program (OHSP),
clinical signs in, 605 Newcastle disease virus (NDV), 293 53
direct examination in, 605, 605f Next-generation sequencing (NGS), for marine bloodborne pathogen safety of, 55
imaging in, 605 mammal viruses, 601 components of, 53
immunology in, 605–606 NGS. see Next-generation sequencing controlled substances in, 54
epidemiology of, 604 NHP. see Nonhuman primate employee training in, 53–54
etiology of, 603–604 Nicotine, embryonic death, in birds, 484–485 ergonomics and, 57
hosts of, 604, 604b Nidovirales, 271 hepatitis B and, 55
phenotype of, 603 Nifurtimox, for Chagas disease, 243 infection control in, 55
postmortem diagnosis of, 604 Nipah virus (NiV), 110–111 sharp objects and, 57
prevention of, 607–608 bats and, 274–275 Occupational health considerations, for elephant
animals, 607 encephalitis from, 275 mycobacteriosis, 670
good staff practices, 607–608 Nipah-like viruses, antibodies against, 275 Occupational health workers, in providing geriatric
water, 607 NMRs. see Naked mole rats zoo animal end of life care, 83–85
treatment for, 606–607 Nobivac D, in nondomestic carnivores, 558t–560t Occupational Safety and Health Administration
Mycobacterium tuberculosis complex (MTBC), 603 Nobivac Puppy-DPv, in nondomestic carnivores, (OSHA), 53
in orangutans, 566t–569t 558t–560t Ocular disease, in Sumatran rhinoceros, 708t
728 Index
Ocular herpes, in chimpanzee, in sanctuary, 577 Ovariohysterectomy, in Hungarian Meadow Viper Pathogens (Continued)
Ocular lesions, of confiscated chelonians, 410f (Vipera ursinii rakosiensis), 391f host-switching, and population-level impacts
O-desmethyl-tramadol (M1), as reptile and Oviposition, 371 of, 100, 100t
amphibian analgesia, 427 Oxymorphone, as reptile and amphibian analgesia, invasion of, 99
Oesophagostomum spp., in orangutans, 566t–569t 424t, 426 reservoirs in, 99
Offshore WEF, 119 Oxyspirura, 476–477 risk analysis for, 101
OHSP. see Occupational Health and Safety Program Oxytetracycline, 407t specialist, 99
Olive Ridley sea turtle (Lepidochelys olivacea), 345 embryonic death, in birds, 484–485 spillover of, 99
Omeprazole, for elephant endotheliotropic Oxytocin, and labor, in elephants, 685–686 vector-borne, 100
herpesvirus, 676t Ozone, in Mycobacterium pinnipedii infection, 607 Pathological fractures, in macropod pediatric
Onderstepoort strain, in nondomestic carnivores, conditions, 503t–504t
561 Patient positioning, on CT table, 214
One Health approach, 94 P PCAB. see Pharmacy Compounding Accreditation
future work of, 96–97 Pacific harbor seals (P. vitulina richardii), brucellosis Board
outcomes and future plans of, 96, 96f in, 312 PCR. see Polymerase chain reaction
project design of, 94–96, 95b, 95f Packaging, 18–19 PCR-based protocols, for marine mammal viruses,
thoughts on, 97 PaCO2, measurement of, in oxygenation, 189–190 600–601
Onshore WEF, 118 PACS. see Picture archiving and communication Pedibothrium spp., in sharks, 341–342
“Open drop” method, inhalant anesthetics delivered system PEDV. see Porcine epidemic diarrhea virus
by, 179 Paecilomyces lilacinus fungal infection, in sharks, 341 Pelecitus, 477
“Open-mouth behavior,” of crocodilians, 415–416 Pain Pelleted feed, for black rhino, 702–703
Operant training, for giraffe lameness, 628 in macropod pediatrics, 502t Pellets, in great ape nutrition, 589–590
Ophidiomyces ophiodiicola, 250–251, 251f, 394 management, in zoological institutions, 151–163 amounts of, based on animal weight, 592t
Ophidiomycosis, 394–397 Pampas deer (Ozotocerus bezoarticus), Babesia spp. in, diet proportions of, by weight, 591t
clinical signs of, 395–396 648t–650t Pelomedusidae, herpesvirus infection in, 270
diagnosis of, 396–397 Pan troglodytes. see Chimpanzees Penicillin, embryonic death, in birds, 484–485
disease outcome of, 395–396 Panamanian golden frogs (Atelopus zeteki), 372 Pentastomidae, 416
distribution and host range of, 394–395 Pancreatic disease, in giraffe, 619 Pentobarbital (sodium)
epidemiology of, 395 Pan-herpesvirus (qPCR) protocols, on equine for amphibians, 361t
etiology of, 394 herpesviruses, 229 for fishes, 360t
pathogenesis of, 395–396 Panthera leo. see Free-ranging lions for reptiles, 361t
treatment of, 397 Papillomaviridae, 269t Peracute mortality syndrome, in giraffe, 619
Opioid antagonists, 165–166 in marine mammals, 600 Perforations, of molars, of elephant, 661
Opioid receptors, in reptile and amphibian, 422 Papillomavirus, 115t Perianesthesia, of musk ox, 638–640, 639f
Opioids, 151–161 Parainfluenza virus, in marine mammals, 599 Perianesthetic monitoring, 185–192
buprenorphine, 153–161 Parainfluenza viruses 1 and 2, in orangutans, of body temperature, 190
in avian, 160–161 566t–569t of cardiovascular system, 185–188
in mammalian, 153–160, 154t–158t Parainfluenza virus 3, in orangutans, 566t–569t arterial blood pressure, 186–188
butorphanol, 161 Paramune 5, in nondomestic carnivores, 558t–560t central venous pressure, 188
in avian, 161 Paramyxoviridae, 269t heart rate, 185
fentanyl, 152–153 in marine mammals, 598–599 heart rhythm, 186
in avian, 153 Paramyxovirus, 115t laboratory parameters in, 190–191
in mammalian, 152, 154t–158t, 160t Paraorygmatobothrium spp., in sharks, 341–342 of pulmonary system, 188–190
in reptilian, 153 Parasites oxygenation, 189–190
for giraffe lameness, 627t–628t in captive Andean bear, 553 SaO2 and PaO2, relationship of, 190
for musk ox, 637 distribution of, due to climate change, 247–249 ventilation, 188–189
receptors of screening, in chimpanzee, 577 Pericoronitis, in elephant, 662
δ or DOP (delta opioid peptide), 151 in sharks, 341 Perimeter fence, 50
κ or KOP (kappa opioid peptide), 151 Parasitic disease and infection, in African wild dog, Periodontal disease, in captive Andean bear, 552,
µ or MOP (mu opioid peptide), 151 542 552f
nociceptin receptor, 151 Parasitology, sample collection, handling, and storage Periodontal probes, in dental disease, of elephant,
Oral disease, in macropod pediatric conditions, in, 202t–203t 660
503t–504t Parelaphostrongylus tenuis, climate change on, 249 Permethrin, for babesiosis, 653
Orangutan hepadnavirus, in orangutans, 566t–569t Parenteral anesthetics, in reptile and amphibian, Persistent corpora lutea, in subfertility associated
Orangutan lymphocryptus virus/Pongine herpes 428 with irregular cyclicity, 127
viruses/Epstein-Barr virus-like, in orangutans, Paromyxovirus, 110–111 Persistent vestibular fistula, in episiotomy, in
566t–569t Partial pulpectomy, for fractured tusks, of elephants, elephants, 686
Orangutans, 565–573 662, 663f Personal safety, 194
air sacculitis in, 570–571, 570t, 593 Partition coefficient, blood to gas, 178b Peste des petits ruminants virus (PPRV), in Saiga
body weights for, 588 Parturition, in elephants, 680–688, 681t antelope, 631–632
diets and digestive physiology of, 588, 589f complications during, 684–685 Pharmacy Compounding Accreditation Board
hepatitis B in, 571 normal, 681–682, 682f (PCAB), 147
infectious diseases in, 565, 566t–569t prediction of, 682–683, 683f–684f Phenol, as treatment for capripoxviruses, 645
tuberculosis in, 571 Parturition-related injury, in subfertility associated 2-phenoxyethanol, for fishes, 360t
vector-borne diseases in, 571 with regular cyclicity, 124–125 Phenylbutazone, for giraffe lameness, 627t–628t
Orbivirus, 115t Parvoviridae, 269t Philasterides dicentrarchi, 332
Organizational influence, 39–44 Passerine birds, systemic isosporosis in, 454–458 Phimosis, in Sumatran rhinoceros, 708t
Orthomyxoviridae, 269t clinical signs and lesions, 454–456, 455f Phlebovirus, 115t
in marine mammals, 598 diagnosis of, 456 Phocid herpesvirus -1 (PHV-1), 599
Orthotic limbs, in avian medicine, 465–470, 466b treatment and management of, 456–457, 456t Physical examination, for diagnosing behavior
candidate and device for, 465–467, 466b Pasteurella multicoda, in Saiga antelope, 632–633 problems, 79–80, 79t
fabrication and care of, 467–469, 468f–469f Pasteurellosis, in Saiga antelope, 630 Physical restraint
OSHA. see Occupational Safety and Health Pasturella spp., in naked mole rats (NMRs), 517 for anteaters, 527
Administration Pathogen pollution, 292 for armadillos, 527
Osmotic pumps, 152 Pathogens for sloths, 527–528
Osteoarthritis, in captive Andean bear, 550–552, distribution of, due to climate change, 247–249 Phytoestrogens, causing cycle irregularity, 127
552f in native wildlife, 99 Picobirnavirus, 115t
Ostertagia greuhneri, climate change on, 250 establishing, 100–101 Picornavirales, 115t
Otarine herpesvirus 1 (OtHV-1), 599 evolution of, 100 Picornaviridae, 269t
Otarine herpesvirus 4, 599 generalist, 99 in tortoises, 270–271
Index 729
Picornavirus, 115t Praziquantel, 407t, 409 Pueblan milk snake (Lampropeltis triangulum
Picture archiving and communication system degradation of, in a recirculating system, 325 campbelli), 394
(PACS), 212 for monogeneans in Spotted eagle rays, 331 Pulmonary system, monitoring of, 188–190
Pinnipeds Precision, definition of, 145 of pulmonary system, 188–190
lens diseases in, 610–617 PREDICT project oxygenation, 189–190
concurrent eye problems and, 611, 611f approach in, 112–113 SaO2 and PaO2, relationship of, 190
incidence of, 610 information management and reporting, 113 ventilation, 188–189
ophthalmologic procedures for, anesthesia in, virus detection and discovery, 112–113 Pulposcopy, in dental disease, of elephant, 660, 661f
612–615 wildlife surveillance at human-wildlife Pulse
outcomes and sequelae in, 615–616 interfaces, 112 in great ape nutrition, 590
risk factors/protective factors for, 610–611 objectives of, 111 palpation of, in monitoring heart rate, 185
surgical candidates for, 615, 615f–616f results of, 113–114, 114f Pulse monitors, in monitoring heart rate, 185
surgical considerations for, 611–612 new models for emergence, 113 Pulse oximeter, in monitoring heart rate, 185
melioidosis in, 317–318 novel viruses, 113, 115t PUREVAX ferret distemper, in nondomestic
Mycobacterium pinnipedii infection in, 603–605, outbreak response, 113–114 carnivores, 558t–560t
607 USAID in, 110–116, 111f Purity, definition of, 145
Pioneer drug, 145 2009-2014, 111, 112f P-value, 26
Plague, in feral cats, 106–107 2014-2019, 115–116 Pyometra, in subfertility associated with regular
Planning Pregnancy, elephant, 680–688 cyclicity, 126
acronyms in, 46t abnormal, 683–684 Pyrazinamide (PZA), for elephant mycobacteriosis,
for geriatric zoo animals, 83–91 normal, 680–681, 681t 666, 670t
for hazards and foreign animal disease, 45–52 Pre-masseteric fossa, of captive Andean bear, 548 Pyrexia, Zika virus infection and, 283
Plaque reduction neutralization test (PRNT), for Premises, designation for foot-and-mouth disease, Python regius, 153
Zika virus infection, 284 48–49, 49t
Plasmodium cynomolgi, in orangutans, 566t–569t Premolars, of elephant, 658
Plasmodium falciparum, in orangutans, 566t–569t Preservation, 47–48 Q
Plasmodium pitheci, in orangutans, 566t–569t Preservation response, 83 Q fever, in camels, 96
Plasmodium silvaticum, in orangutans, 566t–569t Prevalence, in cross-sectional studies, 61 Quality, definition of, 145
Plasmodium spp., in orangutans, 566t–569t Preventive biosecurity, 50 Quality nutrition, in macropod pediatric medicine,
Plasmodium vivax, in orangutans, 566t–569t Primates, melioidosis in, 318 504–505
Platynosomum fastotum, in orangutans, 566t–569t Prion diseases, 255–261, 256t Quality of life assessment, for geriatric zoo animals,
Pleurodira, herpesvirus infection in, 270 clinical signs of, 256–257 83–91
Poaching, Saiga antelope and, 630 diagnostic tools for, 258–259 checklists for, 83, 86f–89f
Poikilotherms, definition of, 357 epidemiology of, 255–256 flow chart for, 85–90, 90f
Poisson models, 26 transmission of, 255–256, 256f models of, 83
Polar bears, encephalitis in, 599 treatment and control of, 259 Quantitative PCR (qPCR), 367
Polio virus, in orangutans, 566t–569t Prions, 255 for elephant mycobacteriosis diagnosis, 665,
Polyenes, for aspergillosis in birds, 441 control of, 259 667t–668t
Polymerase chain reaction (PCR) Priorities, in managing immobilized rhinoceros, 693 Quarantined premises biosecurity, 50
for babesiosis, 652 PRNT. see Plaque reduction neutralization test Quaternary ammonium compound, as treatment for
for Burkholderia pseudomallei, 316 Probe hybridization, for marine mammal viruses, capripoxviruses, 645
for capripoxviruses, 644–645 600–601
for fibropapillomatosis, 398 Problem formulation, 5–6
for ophidiomycosis, 395 Produce, for white rhino, 701 R
Polyomavirus, 115t Progesterone assay, in early embryonic loss, 128 Rabies, 55
in orangutans, 566t–569t Progestin-based melengestrol acetate (MGA) in African wild dog, 542, 545t
Polyopisthocotylea, in saltwater public aquaria, implant, for African wild dog, 541 in cats, 106–107
treatment protocols for, 326t–330t Progestin-based methods, in limiting reproduction, eradication of, 299, 300t–301t
Ponazuril, in systemic isosporosis in Passerine birds, 132 in Madagascar, 294
456t Proparacaine ophthalmic solution, for reptile and in orangutans, 565, 566t–569t
Pongobius spp., in orangutans, 566t–569t amphibian, 424t Raccoon dogs (Nyctereutes procyonoides), SARS CoV
Porcine epidemic diarrhea virus (PEDV), 277 Propofol, 407t and, 276–277
Positive reinforcement, 689–690 for amphibians, 361t Radiation detection bandages, 54–55
Post-Ebola syndrome, 234 for fishes, 360t Radiographs, survey, 218–225
Postmortem tests, for prion disease, 258 for pinnipeds, 613 five basic radiographic opacities in, 218
Postoperative pain relief, for musk ox, 640 for reptiles, 361t future directions of, 223–224
Postpartum complications, in elephants, 687 Prostaglandin E, for elephants, 686 new technology in, 218, 219f–220f
Potassium peroxymonosulfate, for reptiles and Prosthetics, in avian medicine, 465–470, 466b paradigm of
amphibians, 367 candidate and device for, 465–467, 466b next, 221–223
Potency fabrication and care of, 467–469, 468f–469f starting point, 218–221, 221f–224f
definition of, 145 Protein microarray technology, for Middle East planning, summary of, 223
of inhalant anesthetics, 178b respiratory syndrome, 289 Radiography
Potoroids, 494 Protozoa in dental disease, of elephant, 660, 660f
diseases of captive and free-ranging, 498 in Javan rhinoceros, 708t for giraffe lameness, 626
marsupial, conservation translocation of, in saltwater public aquaria, treatment protocols in zoological medicine, 206–207
494–499, 495t for, 326t–330t Radiology, 54–55, 218
conservation management, 494 in Sumatran rhinoceros, 708t Radiology expertise, 213–214
health evaluation and disease surveillance in, Proventricular dilation disease, 461f–462f Rafivirus, 270–271
496 Pseudogymnoascus destructans, 508 Ranaviral disease, in reptiles and amphibians,
postrelease monitoring after, 498 Pseudonitzschia australis, climate change on, 249–250 364–370
restraint, anesthesia, and analgesia in, 495–496, Pseudoparticle virus neutralization tests (ppNTs), for clinical signs and pathology of, 365–366
495f Middle East respiratory syndrome, 289 conservation of, 368
telemetry, 496 Psittacine, bornavirus in, 460 diagnosis of, 366–367
transportation in, 496 Psittacosis, 56, 450b geographic and host distribution of, 364
welfare considerations in, 494–495 Psychiatry, in zoo animals, 79 mortality events of, 368
potential pathogens and parasites of, 497t Public dissections, 29, 30f pathogenesis of, 365
Poxviridae, 269t Public health considerations treatment and prevention of, 367
in marine mammals, 600 for elephant mycobacteriosis, 670 Ranavirus, 293
Poxvirus, 115t related to feral cats, 106–107 Randomized controlled trials, 59
Prairie dogs (Cynomys spp.), vaccination of, 299 Public perception, in surplus animals, 136 Rare diseases, case-control studies for, 61
730 Index
Rash, Zika virus infection and, 283 Reptile ringers, 407t Retroviridae, 269t
Recirculating system, degradation of praziquantel Reptile viruses, 267–273, 269t Retrovirus, 115t
and formalin in, 325 Adenoviridae, 267–269 Rhabdoviridae, 268t–269t
Recombinant viral vector vaccines, in nondomestic arboviruses, 267, 268t Rhabdovirus, 115t
carnivores, 555–557, 558t–560t Arenaviridae, in snakes, 271 Rhesus macaques (Macaca mulatta), MERS-CoV
RECOMBITEK C3, in nondomestic carnivores, Herpesviridae, 269–270 and, 288
558t–560t chelonian, 270 Rhino milk, 700
RECOMBITEK C6, in nondomestic carnivores, crocodilian, 270 Rhino nutrition, 699–706
558t–560t squamate, 270 moving forward, 704
RECOMBITEK canine distemper, 556–557 newly described, 271–272 recommendations for feeding, 699–700,
Record keeping system, for zoo animal welfare, Nidovirales, 271 700t–701t
65–66 Picornaviridae, in tortoises, 270–271 Rhinoceros sondaicus. see Javan rhinoceros
Rectal fluids, for elephant endotheliotropic Reptiles, 359–360 Ribs, of captive Andean bear, 548
herpesvirus, 676t analgesia in, 421–431 Rickettsiosis, 521
Red deer (Cervus elaphus elaphus), Babesia spp. in, local anesthetics as, 428 Rifampicin, for Mycobacterium pinnipedii infection,
648t–650t measurement and quantification of pain and, 607
Red-eared slider turtles (Trachemys scripta elegans), 422 Rifampin
365 multimodal analgesic approaches, 424t–426t, for elephant mycobacteriosis, 666, 670t
Red foxes (Vulpes vulpes), vaccinia-based recombinant 429 for giraffe lameness, 627t–628t
vaccines targeting, rabies and, 299, 300t–301t nonsteroidal anti-inflammatory drugs as, Ringer solution, minimally invasive surgery in
Red panda, vaccine-induced canine distemper in, 428–429 amphibians, 380–381
556t parenteral, 428 Risk
Red-spotted newt (Notophthalmus viridescens), 365 euthanasia methods for, 361t assessment, 6–8
Reference interval studies, 61 fentanyl in, 153 communication, 8–9
Refugia, 335, 335f nociception of pain in, 421–422 defined, 4
Regular cyclicity, subfertility associated with, causes ranaviral disease in, 364–370 management, 8
of, 124–126 clinical signs and pathology of, 365–366 Risk analysis, 4–9
abnormal genitalia in, 124 conservation of, 368 case study, 5–9
age-related changes in, 125 diagnosis of, 366–367 conducting the process, 5
breeding management errors in, 126 geographic and host distribution of, 364 framework, 4–10
male subfertility in, 124 mortality events of, 368 jargon and standards, 4–5
parturition-related injury in, 124–125 pathogenesis of, 365 Risperidone, in chimpanzee, 576
uterine disease in, 126 treatment and prevention of, 367 RMC. see Reproductive Management Center
Rehabilitation medicine Research, naked mole rats (NMRs) in, 517 RNA viruses, 100
of confiscated turtles, 404 Research investigators, definition of, 65 in marine mammals, 597–599
holding conditions of, 408f Research study design, 59–62 Rockborn strain, in nondomestic carnivores, 561
laboratory support of, 411 algorithm for determination of, 59, 60f Rodents, introduced, disease risks from, 295
supplies and equipment for, 406b for evaluation of diagnostic techniques, 61 Roe deer (Capreolus capreolus), Babesia spp. in,
syndromes of, 409–410 diagnostic accuracy studies, 61 648t–650t
triage and initial care of, 405–409, 408t method comparison studies, 61 Root vegetables, in great ape nutrition, 590
of marine turtle, 401 reference interval studies, 61 based on animal weight, 592t
Remifentanil, as reptile and amphibian analgesia, for evaluation of effectiveness of interventions, diet proportions of, by weight, 591t
426–427 59–60 Roping, in immobilization, of capybaras, 522, 522f
Remote injection systems, for African wild dog, 541 crossover trials, 59–60 Rotation, of molars, of elephant, 661, 661f
Renewable energy nonrandomized studies, 60 Rotavirus, 115t
benefits of, 117 randomized controlled trials, 59 Rotavirus SA11, in orangutans, 566t–569t
online, in United States, 117–118 for identification of underlying disease etiologies, Rousettus leishenaulti, novel filoviruses in, 276
sources of, 117 60–61 Rubella virus, in orangutans, 566t–569t
specific energy sources, wildlife impacts from, case-control studies, 61
118–121 cohort studies, 60–61
solar photovoltaic and concentrating solar cross-sectional studies, 61 S
thermal power, 120–121 Reservoirs Safety, vaccination and, 301
wind energy, 118–120 for Ebola virus disease, 235 Safety Data Sheets (SDS), 54
utility-scale, 117 in pathogens, 99 Saiga antelope (Saiga tatarica), mass mortality events
on wildlife, 117–122 Resocialization, of socially deprived chimpanzees, affecting, 630–635, 631t, 632f
Reoviridae, 269t 575 discussion of, 632–633
Reportable avian viruses, 451b Resource-based reviews, for animal welfare Salamanders (Ambystoma tigrinum), 364
Reportable/notifiable disease, necropsy in, 194–195 assessment, 66 Salmonella spp., 57
Reproduction Respiratory disease, in macropod pediatric in orangutans, 566t–569t
of African wild dog, 541, 542f, 542t conditions, 503t–504t Salmonellosis, 450b
of birds, 481 Respiratory syncytial virus, in orangutans, 566t–569t Salt marsh snakes (Nerodia clarkii), 394
of captive Andean bear, 549 Respiratory system, of crocodilians, 415–416 Saltwater aquariums
enhancement of, 130 Respirovirus, in marine mammals, 599 large mixed-species
limiting, 131–132 Reston Ebola virus, in orangutans, 566t–569t capsalid management in, 325
of naked mole rats (NMRs), 514–516, 516b, 516f Reston Ebolavirus (RESTV), 234 leech infestation in, 325–331
Reproductive management, 130–131, 131b Restraint parasites in
changes in, 130–133 of African wild dog, 541 challenges and novel treatment considerations
genetic management vs., 132 of captive Andean bear, 549–550 for, 325–332
lifetime reproductive planning in, 132–133 physical diagnostics of, 323
limiting reproduction in, 131–132 for anteaters, 527 quarantine protocol for, 323, 324b
sustainability crisis in, 130, 131b for armadillos, 527 routine preventive medicine protocol for, 323,
Reproductive Management Center (RMC) for sloths, 527–528 324b
AZA, 130, 131b for sharks, 338–339 techniques for, 323–333
in lifetime reproductive planning, 132 chemical, 339, 339b treatment for, 323–325, 326t–330t
role of, 130 manual, 338–339 Saltwater crocodiles (Crocodylus porosus),
Reproductive pathology, in Sumatran rhinoceros, Results herpesviruses in, 270
712 interpreting, 27 Sample collection, for histology and ancillary
Reproductive system, of crocodilians, 416 statistically significant, 26 diagnostic testing, 201–204, 202t–203t
Reptarenaviruses, inclusion body disease and, 271 summarizing, 26–27 Sample curation, for histology and ancillary
Reptile mite (Ophionyssus natricis), 391, 392f Retinal detachments, in pinnipeds, 616 diagnostic testing, 201–204
Index 731
Sample handling, for histology and ancillary Simian D-type retrovirus, in orangutans, 566t–569t Staphylococcus spp., in orangutans, 566t–569t
diagnostic testing, 201, 202t–203t Simian foamy virus, in orangutans, 566t–569t Starch, in great ape nutrition
San Miguel sea lion virus (SMSV), 598 Simian T-lymphotropic virus-1, in orangutans, amounts of, based on animal weight, 592t
Sanctuary, chimpanzee in, 574–580 566t–569t diet proportions of, by weight, 591t
behavioral issues in, 578 Sindbis virus, in orangutans, 566t–569t Starvation, of confiscated turtles, 409
demyelinating disease in, 577–578 Skin disease, in macropod pediatric conditions, State Animal Health Officials, 46t
end of life and euthanasia for, 578–579 503t–504t State of conservation, of capybaras, 520
housing of, 575–576 Skin syndromes, of confiscated turtles, 410 Statistical analysis, data structure, 24, 25f
ocular herpes in, 577 Sleep behavior, in diagnosing behavior problems, Statistical test/method, 25–26, 25b–26b
physical examinations and anesthesia for, 576 77–78 Statistics in wildlife studies, practical guide for,
preventative healthcare, 577 Sloths 21–27
rehabilitation of, 574–580 anesthetic monitoring in, 533 Stem cells, 138–139
reproductive management in, 577 anesthetic recovery in, 533 in clinical medicine, 140–141
status of, 574–575 chemical immobilization and anesthesia in, collection of, 139–140
SaO2, measurement of, in oxygenation, 190 529t–531t, 532–533 definition of, 138
Sarcocystosis, 447b physical restraint for, 527–528 mechanism of, 139
Sarconema eurycerca, 477 Small animals, necropsies on, 199 in zoo medicine, 138–144
SARS. see Severe acute respiratory syndrome “SMART” format, 35, 35b Steppe viper (Vipera renardi), 389
Saturated vapor pressure, 178b SMSV. see San Miguel sea lion virus Stereotypical behavior, in animal welfare assessment,
Scrapie, 256t Snake fungal disease (SFD), 250–251, 251f, 394 68
atypical, 256t Snakes, Arenaviridae in, 271 Stereotypies, in captive Andean bear, 549
control of, 259 Snyder Hill strain, in nondomestic carnivores, 561 STIV. see Soft-shelled turtle virus
transmission of, 255 Social environment, in behavior assessment, 77–78 Strategic objectives, 34–37
Scuticociliatosis Social housing, of chimpanzee, in sanctuary, 575 Strategic priorities, 35–37
in saltwater public aquaria, treatment protocols Social structure, of giraffe, 620 Streptocara incognita, 471, 476f
for, 326t–330t Sodium dodecyl sulfate, as treatment for Streptococcus spp., in orangutans, 566t–569t
in Sygnathid and Elasmobranch species, 332 capripoxviruses, 645 Stress, animal welfare and, endocrinological
SDS. see Safety Data Sheets Sodium hypochlorite evaluation of, 73–75
Sea ice, loss of, due to climate change, 250 for reptiles and amphibians, 367 overview of stress response, 73, 74f
Sea star wasting disease (SSWD), 250, 251f as treatment for capripoxviruses, 645 types of samples for glucocorticoid assessment,
Sea turtles Soft-shelled turtle virus (STIV), 364 73–74
decompression medicine in, 345–350 Solar facilities (SF), 120 validation procedures for, 74
herpesvirus infection in, 270 Solar photovoltaic and concentrating solar thermal Strongylida spp., in orangutans, 566t–569t
Seal bacillus. see Mycobacterium pinnipedii power, wildlife impacts from, 120–121 Strongyloides fuelleborni, in orangutans, 566t–569t
Secure Zoo Strategy, foot-and-mouth disease South American sea lion, Mycobacterium pinnipedii Strongyloides spp., in orangutans, 566t–569t
incident annex using, 48–51, 49t, 50f infection in, 604 Strongyloides stercoralis, in orangutans, 566t–569t
Sedation, musk Ox, 636–640 Southeast Asia, elephant care in, 689–691 Student t-test, 26
Seeds, in great ape nutrition, 590 captivity of, 689 Study
Selection bias, 22 handling of, 689 sample size, 22–23
Selenium, embryonic death, in birds, 484–485 history of, 689 validity, 22, 23f
Sepsis, in macropod pediatrics, 502t Target Training Project and, 689–691, 690f–691f, Subcutaneous Ophidiomyces ophiodiicola mycetoma,
Septicemia, of confiscated turtles, 409 691b 396f
Serologic assays, in reptiles and amphibians, 367 Southern sea otter (Enhydra lutris nereis), brucellosis Subfertility, causes of
Serology in, 310 associated with early embryonic loss, 128
in babesiosis, 652–653 Spawning, 371 associated with irregular cyclicity, 126–127
for elephant mycobacteriosis diagnosis, 665–666, Specialist pathogens, 99 age-related changes, 127
667t–668t Species Survival Plan Programs (SSP), 194 anovulatory follicles, 127
Serratospiculosis, 477–478, 478f Specific heat, of vaporization, 178b endocrine disorders, 127
Serum biochemistry, of crocodilians, 414–415 “Spectacled (Andean) bear alopecia syndrome, in intersex conditions, 126–127
Setaria tundra, climate change on, 249 captive Andean bear, 552 persistent corpora lutea, 127
Severe acute respiratory syndrome (SARS), 110–111 Spheniscus humboldt, 214 poor nutrition, 127
bats and, 274, 276–277 Spinal cord severance, for reptiles, 361t associated with regular cyclicity, 124–126
in reptiles, 271 Spinal deformities, in sharks, 342 abnormal genitalia, 124
Sexual maturity, of captive Andean bear, 549 Spirocerca lupi, in lemurs, 295 age-related changes, 125
SFD. see Snake fungal disease Spirurida spp., in orangutans, 566t–569t breeding management errors, 126
Shark box, 339 Spirurids, 471, 472t male subfertility, 124
Sharks avian, 471–480, 474t–475t parturition-related injury, 124–125
biology, anatomy, and physiology of, 338 diplotriaenoid, 477–478, 478f uterine disease, 126
diagnostics of, 339–340, 340f Spoligotyping Suburban ecosystems, feral cats in, 106
diseases of, 341–343 in MTBC, 605, 605f Suckling, in neonatal macropods, 500
husbandry and management of, 338 in Mycobacterium pinnipedii infection, 603–605, Sudan Ebolavirus (SUDV), 233
imaging of, 340 605f Sudden death, in macropod pediatric conditions,
medicine and, 338–344 Spongiform encephalopathy, type-H, atypical, 256t 503t–504t
physical examination of, 339 Spotted eagle rays, monogeneans in, 331 Sugars, for white rhino, 701
physiologic characteristics of, 339–340, 341t–342t Spotted turtles (Clemmy guttata), herpesvirus Sulawesi tortoises, 267
restraint for, 338–339 infection in, 270 Sulfachlorpyrazine, in systemic isosporosis in
chemical, 339, 339b SPPV. see Sheeppox virus Passerine birds, 456t
manual, 338–339 Spray exposure, by zoo and wildlife veterinarians, Sulfachlorpyridazine, in systemic isosporosis in
treatment and therapies for, 343 167t–168t, 169 Passerine birds, 456t
Sharp objects, OHSP and, 57 Squamates, herpesviruses in, 270 Sulfadiazine trimethoprim, for giraffe lameness,
Sheeppox virus (SPPV), 641 SSWD. see Sea star wasting disease 627t–628t
Shell lesions, of confiscated chelonians, 410f Staff, on geriatric zoo animals, 83–85 Sulfadimethoxine, in systemic isosporosis in
Shell syndromes, of confiscated turtles, 410 Stakeholders Passerine birds, 456t
Shigella spp., in orangutans, 566t–569t in controversy, over feral cats, 105 Sumatran rhinoceros, 707–712
Shipping, port of entry and, 19 in providing geriatric zoo animal end of life care, infectious and emerging disease in, 707–710
Siadenoviruses, 267 83, 85t bacteria, viruses, and preventative medicine,
Siamese crocodile (Crocodylus siamensis), ultrasound Standard equipment, of minimally invasive surgery, 707, 709t
and gross appearance of spleen and steatotheca 381b endoparasites, 709–710
of, 415f Standing sedation, for elephant endotheliotropic tabanids and trypanosomes, 710
Sidestream analyzers, 189 herpesvirus, 676t ticks and tick-borne disease in, 710
732 Index
Sumatran rhinoceros (Continued) Tetracycline, embryonic death, in birds, 484–485 Transmissible mink encephalopathy (TME),
noninfectious disease in, 710–712 Tetrameridosis, 471–476, 476f–477f 256–257, 256t
chronic renal disease, 712 Tetraphyllidean spp., in sharks, 341–342 Transmissible spongiform encephalopathies (TSEs).
eye disorders, 710–711 Theileria bicornis, in Sumatran rhinoceros, 710 see Prion diseases
iron storage disease, 711 Theileria spp., in Sumatran rhinoceros, 710 Transmission electron microscopy (TEM), of
reproductive pathology and allee effect, 712 Thelazia, 476–477, 477f fibropapillomatosis, 398
records of disease outbreaks in, 708t Therapy laser, for giraffe lameness, 627t–628t Trap-Neuter-Release (TNR), in cats, 104–105
taxonomy of, 707 Thermography, for giraffe lameness, 626 management alternatives to, 107
Sunshinevirus, in reptiles, 271–272 Thiafentanil (Thianil) Trauma, of giraffe, 619–621, 620f
Sunviridae, 269t in accidental veterinary anesthetic exposure, Traumatic fractures, in macropod pediatric
in reptiles, 271–272 169–171 conditions, 503t–504t
Supernumerary molars, of elephant, 661 in zoo and wildlife medicine, 165 Trawler (Crespo-Picazo), 345
Supernumerary tusks, of elephant, 661 THIRA. see Threat and Hazard Identification and Tremarctos ornatus. see Andean bear
Suprelorin (deslorelin acetate) implants, for African Risk Assessment Triamcinolone, for elephant endotheliotropic
wild dog, 541 Threat and Hazard Identification and Risk herpesvirus, 676t
Surgical lensectomy, for cataracts, in pinnipeds, Assessment (THIRA), 46t, 47 Tricaine methanesulfonate
611–612 Ticarcillin, 407t for amphibians, 361t
Surplus animals Tick-borne disease for reptiles, 361t
breeding in, 135 climate change on, 248 Trichechid herpesvirus 1 (TrHV-1), 599
dealing with, 135–136, 135f in Sumatran rhinoceros, 710 Trichlorfon, for leech infestation, 325–331
definition of, 134 Ticks, in Sumatran rhinoceros, 710 Trichodina spp., in saltwater public aquaria,
issues surrounding, in zoos, 134–136 Tigers (Panthera tigris), conservation of, vaccines for, treatment protocols for, 326t–330t
meeting the demand, 134–135 302–303 Trichomonas, in orangutans, 566t–569t
public perception in, 136 Tiletamine-zolazepam Trichomoniasis, 450b
sustainable populations in, 134, 134f for captive Andean bear, 550t Trichophyton spp., in orangutans, 566t–569t
Surveillance, in white-nose syndrome, 511 for chimpanzee, 576 Trichostrongylus spp., in orangutans, 566t–569t
Survey radiographs, 218–225 for free-ranging lions, 536–537, 537t Trichuris spp., in orangutans, 566t–569t
five basic radiographic opacities in, 218 for xenarthrans, 529t–531t Troponins, for great ape cardiovascular disease,
future directions of, 223–224 Tiletamine-zolazepam ketamine, for captive Andean 585–586
new technology in, 218, 219f–220f bear, 550t Trypanosoma cruzi, 239–246
paradigm of Tiletamine-zolazepam-medetomidine background of, 239, 240f
next, 221–223 for musk ox, 637 diagnosis of, 243f, 244
starting point, 218–221, 221f–224f for xenarthrans, 529t–531t epidemiology of, 239–242
planning, summary of, 223 Time use, in managing immobilized rhinoceros, genetics of, 241–242
Suspect premises, 49t 693 hosts, 239–241
Sustainability, in surplus animals, 134, 134f Tissuevax, in nondomestic carnivores, 558t–560t transmission routes, 241
Sustainability crisis, in animal populations, 130 TME. see Transmissible mink encephalopathy vectors, 241, 241f
Sustained-release (SR), opioid drugs, 152, 154t–158t TNR. see Trap-Neuter-Release management of, 243–244, 244b
anecdotal reports of, 152, 160t Togaviridae, 268t–269t pathogenesis of, 242–243
SWOT analysis, 36, 37b, 37f Tomato frogs (Dyscophus spp.), 372 clinical signs, 242
Sygnathids, scuticociliatosis in, 332 Tonic immobility, 338–339 pathology, 242–243, 242f
Systematic approach, in diagnosing behavior Topicals, for giraffe lameness, 627t–628t prevention of, 243–244
problems, 76–82 Topivirus, 270–271 treatment of, 243–244
applied behavior analysis, 76–77 Tortoises zoonotic potential of, 244
comprehensive behavioral history, 77–78 herpesvirus infection in, 270 Trypanosoma evansi
steps of, 76, 77f Picornaviridae in, 270–271 blood film evaluation of, in camels, 96, 96f
thorough physical examination, 79–80, 79t Touch-pools, 334–337 in Sumatran rhinoceros, 708t
Systemic isosporosis, in passerine birds, 454–458 animal health and welfare criteria in, 336–337 Trypanosoma lewisii, in Madagascar, 295
clinical signs and lesions, 454–456, 455f contingency planning for, 336 Trypanosoma spp., in orangutans, 566t–569t
diagnosis of, 456 feeding and behavior records in, 336 Trypanosomes, in Sumatran rhinoceros, 710
treatment and management of, 456–457, 456t hand-washing and drying stations in, 336 TSA. see Turtle Survival Alliance
holding space in, 336 Tuberculin injection, for Mycobacterium pinnipedii
life-support system of, 335–336 infection, 606
T monitoring of, 336–337 Tuberculin skin test, for elephant mycobacteriosis
Tabanids, in Sumatran rhinoceros, 710 planning of, 335–336, 335f diagnosis, 667t–668t
Tachycardia, 186 setting goals for, 334 Tuberculosis, in orangutans, 571
TAG. see Taxon Advisory Group species selection for, 334–335 Tulathromycin, 407t, 409
Taï Forest Ebolavirus (TAFV), 233 staff and guest considerations in, 337 for giraffe lameness, 627t–628t
Tapentadol, as reptile and amphibian analgesia, water quality of, 336 Tumors, in molars, of elephant, 661
425t–426t, 428 Toxicology, sample collection, handling, and storage Turtle Survival Alliance (TSA), 404
Target Training Project, 689–691, 690f–691f, 691b in, 202t–203t Tushes, 657–658
Tasmanian devil facial tumor disease, 490–493 Toxoplasma gondii Tusklets, of elephant, 662
cause of, 490–491 in cats, 106–107 Tusks, of elephant, 658–659, 658f
devil populations, 491–492 lemurs and, 295 abnormal structure of, 661
moving forward, 492, 492f Toxoplasmosis, 56 absent, 661
prevention of, vaccines for, 302–303 Trachemys scripta, 214 diseases/anomalies affecting, 661–662
signs and symptoms of, 490, 491f Training techniques, for giraffe, 621 treatment of, 662, 663f
“Tattoo skin lesions”, 600 Tramadol, 407t fractures of, 657, 658f, 662
Taxon Advisory Group (TAG), 194 for giraffe lameness, 627t–628t supernumerary, 661
T-cell lymphotropic/leukemia virus type 1 (STLV-1), as reptile and amphibian analgesia, 427
in orangutans, 565 Tranquilizers, long acting, use of, 638
Telazol, for African wild dog, 541 Transdermal fentanyl delivery systems or patches, U
Telemetry, for potoroids, 496, 496f 152 Ultra-potent opioids (UPOs), used in zoo and
Teleosti (bony fishes), 357 in avian, 153 wildlife anesthesia, 164–176
TEM. see Transmission electron microscopy in mammalians, 152 alpha-2 agonists and antagonists, 166
Tembusu virus, in orangutans, 566t–569t in reptilian, 153 antagonists and, 164–165
Tenacibaculum maritimum, sharks and, 341 Transdermal fentanyl solution potency comparisons of, 165, 165t
Terbinafine, as antifungals in birds, 442t, 444 in avian, 153 Ultrasound
Testadenoviruses, 267–269 in mammalians, 152 for crocodilians, 413, 414f–415f
Testudinid herpesvirus, 270 Transferrin saturation, in Sumatran rhinoceros, 711 for Mycobacterium pinnipedii, 605
Index 733
Ultrasound (Continued) Venomous snakes, necropsies on, 199 Weight loss diet, for obesity in great ape, 593
for sea turtles, 346, 347f–349f Ventilation, in sharks, 338 Welfare, of zoo animals, 64–72
for sharks, 340 Ventilometry, in ventilation, 189 assessments of, 64–66
Ultraviolet (UV) devices, for MTBC, 607 Ventral coccygeal vein, of sharks, blood collected practical guidelines for, 64–66
Ultraviolet (UV) Index, in lens diseases, in from, 339, 340f principles for, 66–68, 67t
pinnipeds, 611 Vero cells, in nondomestic carnivores, 561 committees for, 68–70, 69t
Ultraviolet (UV) light, lens diseases and, in Versatility, computed tomography provides, 207, departments for, 70
pinnipeds, 611 209f introduction to, 64
Umbilical cord, for stem cell collection, 139–140 Vesicular exanthema of swine virus (VESV), 598 veterinarian’s role in, 70
Umingmakstrongylus pallikuukensis, climate change VESV. see Vesicular exanthema of swine virus West African mud turtles (Pelusios castaneus),
on, 248 Veterinarians, 65 herpesvirus infection in, 270
UMRVs. see Unclassified Morbilli-related in feral cat, 107–108 West Nile virus (WNV), 293
paramyxoviruses opportunities for, 28–33 climate change on, 250
Unclassified Morbilli-related paramyxoviruses booths, 28, 29f in Sumatran rhinoceros, 707
(UMRVs), in Madagascar, 295 education programming, 29–30 Western lowland gorillas (Gorilla gorilla gorilla)
Undulant fever, 306 Little Zoo Vets program, 30–32, 31f–32f, 31t Ebola virus disease on, 233
United States Agency for International Development mentorship, 28–29 human T-lymphotropic virus in, 113
(USAID), Emerging Pandemic Threats public dissections, 29, 30f Wharton jelly, for stem cell collection, 139–140
PREDICT project, 111, 112f veterinary windows, 29, 29f White hyphae, in white-nose syndrome, 510, 510f
United States Department of Agriculture (USDA), role in zoo animal welfare, 70 White-nose syndrome, 508–513
46t Veterinary Advisors clinical signs of, 509–510, 510f
UPOs. see Ultra-potent opioids background of, 2, 3t diagnosis of, 510–511, 510f
Urban ecosystems, feral cats in, 106 responsibilities, 2–3 disease mitigation, 511
Urinary system, of crocodilians, 416 role of, 2–3 global perspective, 508
Urolithiasis, in giraffe, 619 Veterinary import permits, 17–18 interdependence, host, and environment,
Uronemia spp., in saltwater public aquaria, treatment Veterinary occupational health and safety, in zoo and 508–509
protocols for, 326t–330t wildlife setting, 53–58 North American perspective, 508, 509f
US Fish and Wildlife Service (USFWS), 117 Veterinary Safety Manual (VSM), 54 surveillance in, 511
US Pharmacopeial Convention (USP), 147 Veterinary team, in providing geriatric zoo animal White rhinoceros
USAID. see United States Agency for International end of life care, 83–85, 85t field immobilization of
Development (USAID) Veterinary windows, 29, 29f drugs for, 692, 693t
USDA. see United States Department of Agriculture Vibrio carchariae, sharks and, 341 managing in, 693–695, 694t
USFWS. see US Fish and Wildlife Service (USFWS) Vibrio damsela, sharks and, 341 reversal of, 695–696
Uterine disease, in subfertility associated with regular Video recordings, for behavioral analysis, 76–78 hyperthermia in, 695
cyclicity, 126 Video systems, 13 monitoring breathing and response to hypoxemia
Uterine inertia, causing dystocia, in elephants, Viral erythrocytic necrosis, in sharks, 341 and apnea, 694
684–685 Virkon, as treatment for capripoxviruses, 645 monitoring heart rate and blood pressure in, 695
Uterine pathology, in Sumatran rhinoceros, 708t Virology muscle tremors in, 695
Uterine trauma, in fetotomy, in elephants, 686 of avian influenza, 262–263 nutrition, 701, 702t
Utility-scale concentrating solar power project, 118 sample collection, handling, and storage in, off-loading, 697
Utility-scale renewable energy, 117 202t–203t transporting, 695f, 696–697
Utility-scale solar PV plant, 118 Virus isolation, for marine mammal viruses, 600 airlifting, 696, 696f
Utility-scale wind farm, 118 Viruses orphan calves, 697
classification of, 597, 598t transporting by vehicle, 696
detection and discovery of, in PREDICT walking of, 695
V approach, 112–113 White-tailed deer (Odocoileus virginianus), Babesia
Vaccinated premises, 49t DNA, 599–600 spp. in, 648t–650t
Vaccination in marine mammal, 597–602 Whole grains, in great ape nutrition, 590
for Ebola virus disease, 235–236 novel, 113, 115t Wilcoxon rank sum test, 26
for equine herpesviruses, 231 RNA, 597–599 Wild bog turtles (Glyptemys muhlenbergii),
in macropod pediatric medicine, 505 in wildlife zoonotic disease emergence, 110 herpesvirus infection in, 270
for nondomestic avian species, against avian Vision, in strategic planning, 34, 35b, 36 Wild carnivores, brucellosis in, 309–310
influenza viruses, 265 Visitation, 48 Wild pigs, brucellosis in, 308–309
of wildlife areas for, 50 Wildlife
cautionary principles in, 302 Vitamin D3, in great ape nutrition, 590 climate change on disease spread in, 247–254
delivery considerations for, 301–302 Vitamin E, in white rhino, 701 distribution of pathogens, parasites, and
delivery methods for, 301 Vitamin supplementation, in great ape nutrition, vectors, 247–249, 248f
efficacy of, 302 590 emerging diseases, 250–251, 251f
future of, 302–303 Voriconazole, as antifungals in birds, 442t, 443 prevalence or severity of disease, 249–250, 249f
motivations for, 299–301, 300t–301t VSM. see Veterinary Safety Manual recommendations on, 251–252, 252b
safety of, 301 compounding relevance and considerations for,
techniques for, 299–305 148
for zoo employees, 55, 56t W prion diseases in, 255–261, 256t
Vaginal strictures, in subfertility associated with Walk-through aviaries, medical management of, clinical signs of, 256–257
regular cyclicity, 124 446–453 diagnostic tools for, 258–259
Vaginal vestibulotomy, for elephants, 686 animal sourcing and quarantine in, 449 epidemiology of, 255–256
Valvular dysplasia, in African wild dog, 542–543 exhibit design of, 446–447, 447b, 447f, 448t transmission of, 255–256, 256f
Vanguard, in nondomestic carnivores, 558t–560t flock nutrition and husbandry in, 448–449 treatment and control of, 259
Vanguard D, in nondomestic carnivores, 558t–560t screening for zoonotic agents in, 449, 450b renewable energy on, 117–122
Vapor pressure, 178b species choice, 446 from specific energy sources, 118–121
Vaporizers, for free-ranging wildlife, 177–184, 178b Water solar photovoltaic and concentrating solar
Varicella zoster virus, in orangutans, 566t–569t for equine herpesviruses transmission, 230–231 thermal power, 120–121
Vascular access, in pinnipeds, 613 for great ape nutrition, 588–589 wind energy, 118–120
Vasectomies, in chimpanzee, 577 Water buffalo, stem cell collection in, 139–140 Wildlife Conservation Society (WCS), 404
Vector-borne diseases, in orangutans, 571 Water quality Wildlife disease risk, 5
Vector-borne pathogens, 100 monitoring of, in touch-pools, 336 Wildlife health concerns, related to feral cats,
Vectors, distribution of, due to climate change, for sharks, 338 106–107
247–249 Water-bath-based anesthesia, of amphibians, 386 Wildlife hosts, of Mycobacterium pinnipedii, 604,
Vegetables, in great ape nutrition, 590 WCS. see Wildlife Conservation Society 604b
diet proportions of, by weight, 591t WEF. see Wind energy facilities (WEF) Wildlife interface, 5
734 Index
Wildlife necropsy primer, 194–205 Xenogeneic mesenchymal stem cells, 142 Zoo veterinary operations, strategic planning for,
basics of, 194–196, 195b, 197f Xenogeneic relationship, in mesenchymal stem cells, 34–38
examination of, 196–204 139 components of, 34–35, 35b, 38f
communication during, 204 Xylazine creating a, 35–38
data management and sharing in, 204 for captive Andean bear, 550t importance of, 34
external, 198, 198b in zoo and wildlife medicine, 166 Zookeepers, in providing geriatric zoo animal end of
internal, 198, 198b Xylazine butorphanol midazolam, for xenarthrans, life care, 83–85, 85t
relevant historical information, collection and 529t–531t Zoological Information Management System
documentation in, 196 Xylazine-ketamine, for musk ox, 637 (ZIMS), 412
sample collection and management in, Zoological medicine
201–204, 202t–203t computed tomography in, 206–217
Wildlife surveillance, at human-wildlife interfaces, in Z benefits of, 206–208
PREDICT approach, 112–113 Zaire Ebolavirus (ZEBOV), 233–235 equipment considerations of, 208–210
Wildlife technologies, 11–15 “Zero risk”, 5 interventional, 215, 216f
biologging, 11–12 Zika virus, 280–286 optimization of, 214–215
biotelemetry, 11–12 final considerations in, 284–285 review of, 212–214
digital imaging, 13 infection with magnetic resonance imaging in, 206–217
environmental loggers, 12–13 clinical manifestations of, 283, 285f benefits of, 206–208
Wildlife zoonoses, global detection of, 110–116 complete blood count and blood chemistry equipment access to, 208–209
Wind energy panels for, 283 review of, 212–214
as renewable energy, 117 diagnosis of, 283 sustained-release and long-acting opioid
wildlife impacts from, 118–120 epidemiology and wild hosts of, 280–282 formulations of, 151–163
Wind energy facilities (WEF), 118 necropsy in, 284 buprenorphine, 153–161
Wing membranes, in white-nose syndrome, 510 pathogenesis of, 282–283 butorphanol, 161
Winter ticks, climate change on, 249 treatment, prevention, and control of, fentanyl, 152–153
WNV. see West Nile virus 284 Zoonotic viruses, 274
Wood frogs (Lithobates sylvaticus), 364 investigation of, collecting biologic materials for, Zoos
Wood turtles (G. insculpta), herpesvirus infection 283 animal welfare in, 64–72
in, 270 molecular, 280 assessments of, 64–66
Woodland caribou (Rangifer tarandus caribou), in orangutans, 566t–569t committees for, 68–70, 69t
Babesia spp. in, 648t–650t organs targeted by, 283 departments for, 70
Work tracking plant species, for Indian rhino ZIMS. see Zoological Information Management veterinarian’s role in, 70
nutrition, 702 System compounding relevance and considerations for,
World, European, or Association of Zoos and Zolazepam-tiletamine (ZT), for captive Andean bear, 148
Aquariums (WAZA, EAZA, AZA), 194 549–550 definition of, 64, 65t
World Courier, 19 Zoo animals disease risks to, 99–103
World Organization for Animal Health, on age-related health conditions in, 83, 84f–85f, pathogens in, 99–100, 100t
definition of animal welfare, 65t 84t risk analysis for, 101–103, 102t
Wyoming toads (Anaxyrus baxteri), 372 geriatric One Health approach and, 94
end of life planning for, 83–91 future work of, 96–97
quality of life assessment for, 83–91 outcomes and future plans of, 96, 96f
X longevity of, 85 project design of, 94–96, 95b, 95f
Xenarthra, 527–534 prion disease in, 256–257 thoughts on, 97
chemical immobilization and anesthesia for, Zoo medicine surplus animals in, 134–136
528–533 mesenchymal stem cells in, 141–142
physical restraint in, 527–528 stem cell therapy in, 138–144
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