Animal Models for the Study of Human

Disease 2nd Edition P. Michael Conn

Visit to download the full and correct content document:
https://textbookfull.com/product/animal-models-for-the-study-of-human-disease-2nd-e
dition-p-michael-conn/
More products digital (pdf, epub, mobi) instant
download maybe you interests ...

Neurobiology of Disease 2nd Edition Michael V Johnston

Editor Harold P Adams Editor Ali Fatemi Editor

https://textbookfull.com/product/neurobiology-of-disease-2nd-
edition-michael-v-johnston-editor-harold-p-adams-editor-ali-
fatemi-editor/

Conn's handbook of models for human aging 2nd Edition

Ram

https://textbookfull.com/product/conns-handbook-of-models-for-
human-aging-2nd-edition-ram/

Infectious Disease Management in Animal Shelters 2nd

Edition Lila Miller

https://textbookfull.com/product/infectious-disease-management-
in-animal-shelters-2nd-edition-lila-miller/

Animal movement : statistical models for animal

telemetry data 1st Edition Mevin B. Hooten

https://textbookfull.com/product/animal-movement-statistical-
models-for-animal-telemetry-data-1st-edition-mevin-b-hooten/
Computer Simulation of Liquids 2nd Edition Michael P.
Allen

https://textbookfull.com/product/computer-simulation-of-
liquids-2nd-edition-michael-p-allen/

The innate immune response to non-infectious stressors

: human and animal models 1st Edition Amadori

https://textbookfull.com/product/the-innate-immune-response-to-
non-infectious-stressors-human-and-animal-models-1st-edition-
amadori/

Animal Models of Acute Neurological Injury Jun Chen

https://textbookfull.com/product/animal-models-of-acute-
neurological-injury-jun-chen/

Contract Formation: Law and Practice 2nd Edition

Michael P. Furmston

https://textbookfull.com/product/contract-formation-law-and-
practice-2nd-edition-michael-p-furmston/

Animal Ethos The Morality of Human Animal Encounters in

Experimental Lab Science First Edition Lesley A. Sharp

https://textbookfull.com/product/animal-ethos-the-morality-of-
human-animal-encounters-in-experimental-lab-science-first-
edition-lesley-a-sharp/
ANIMAL MODELS
FOR THE STUDY
OF HUMAN
DISEASE
Second Edition
Dedicated to the memory of P. Michael Conn
ANIMAL MODELS
FOR THE STUDY
OF HUMAN
DISEASE
Second Edition

Edited by

P. Michael Conn
Senior Vice President for Research
Associate Provost and Professor of Internal Medicine and Cell Biology-Biochemistry
Texas Tech University Health Sciences Center, Lubbock, TX, United States
Academic Press is an imprint of Elsevier
125 London Wall, London EC2Y 5AS, United Kingdom
525 B Street, Suite 1800, San Diego, CA 92101-4495, United States
50 Hampshire Street, 5th Floor, Cambridge, MA 02139, United States
The Boulevard, Langford Lane, Kidlington, Oxford OX5 1GB, United Kingdom

Copyright © 2017 Elsevier Inc. All rights reserved.

No part of this publication may be reproduced or transmitted in any form or by any means, electronic or
mechanical, including photocopying, recording, or any information storage and retrieval system, with-
out permission in writing from the publisher. Details on how to seek permission, further information
about the Publisher’s permissions policies and our arrangements with organizations such as the Copy-
right Clearance Center and the Copyright Licensing Agency, can be found at our website: www.elsevier.
com/permissions.

This book and the individual contributions contained in it are protected under copyright by the Publisher
(other than as may be noted herein).

Notices
Knowledge and best practice in this field are constantly changing. As new research and experience broaden
our understanding, changes in research methods, professional practices, or medical treatment may become
necessary.

Practitioners and researchers must always rely on their own experience and knowledge in evaluating and
using any information, methods, compounds, or experiments described herein. In using such information
or methods they should be mindful of their own safety and the safety of others, including parties for whom
they have a professional responsibility.

To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors, assume any
liability for any injury and/or damage to persons or property as a matter of products liability, negligence
or otherwise, or from any use or operation of any methods, products, instructions, or ideas contained in the
material herein.

Library of Congress Cataloging-in-Publication Data

A catalog record for this book is available from the Library of Congress

British Library Cataloguing-in-Publication Data

A catalogue record for this book is available from the British Library

ISBN: 978-0-12-809468-6

For information on all Academic Press publications visit our website at

https://www.elsevier.com/books-and-journals

Publisher: Sara Tenney

Editorial Project Manager: Fenton Coulthurst
Production Project Manager: Lucía Pérez
Designer: Greg Harris

Typeset by Thomson Digital

 Dedication
I began working for Dr. P. Michael Conn in his research
laboratory in September 1984, and had continued to work
with him until his untimely passing in November 2016.
Dr. Conn was a superb mentor, friend, and human being.
He mentored numerous postdoctoral fellows, graduate
students, and staff throughout his research career, and,
in turn, many of the fellows and students have success-
ful careers and hold Dr. Conn in very high regard.
Dr. Conn was known for his research involving the
mechanisms of action of gonadotropin-releasing hor-
mone in the pituitary, and therapeutic approaches that
restore misfolded protein functioning. He authored
and coauthored over 350 publications in his field and
wrote and edited over 200 books, including texts in
neurosciences, molecular biology, and endocrinol-
ogy. He was recognized with a MERIT Award of the
National Institutes of Health; the J.J. Abel Award of
the American Society for Pharmacology and Experi-
mental Therapeutics; the Weitzman, Oppenheimer, and
Ingbar Awards of the Endocrine Society; the Miguel
Aleman Prize; Mexico’s National Science Medal; and
the Stevenson Award of Canada. He was the recipient
of the Oregon State Award for Discovery, the Media
Award of the American College of Neuropsychophar-
macology, and was named distinguished Alumnus of
Baylor College of Medicine in 2012. Dr. Conn served as
the editor of numerous professional journals and book
series: Endocrinology, Journal of Clinical Endocrinology
and Metabolism, Endocrine, Methods, Progress in Molecular
Biology and Translational Science, and Contemporary
Endocrinology. He was an advocate for the value of ani-
mal research and the dangers posed by animal extrem-
ism. He was the coauthor of The Animal Research War
(2008) and wrote many opinion editorials for the pub-
lic and academic community, which appeared in The
Washington Post, The LA Times, The Wall Street Journal,
the Des Moines Register, and elsewhere.
I am honored that I spent 32 years learning and work-
ing with Dr. Conn, a true friend.
Dr. P. Michael Conn will be sorely missed. Rest in
peace my friend!
Jody Janovick
Texas Tech University Health Sciences Center,
Lubbock, TX, United States
Page left intentionally blank
 Contents

List of Contributors  xv 3 Animal Models in Biomedical Research 73

4 Concerns About the Translatability of Findings
Preface  xix
From Animal Experimental Studies 73
5 Translational Research 73
Part A 6 Choice of Appropriate Animal Model
7 Where in the Process of Modeling Human Diseases
75

ETHICS, RESOURCES, AND and Developing Putative Therapeutics Have Large

APPROACHES Animal Models Been Used? 77
8 Which Model Animal Species Are Classified as Large
in Scientific Research? 78
1. Ethics in Biomedical Animal Research: The Key 9 Which Types of (Large) Animal Models
Role of the Investigator Are Available? 79
JERROLD TANNENBAUM 10 Special Aspects in Using Large Farm Animal Models 80
11 Experimental Unit 81
1 Nature and Scope of the Chapter 4 12 Experiments Using Social Animals Requiring
2 The Subject Matter of Biomedical Animal Research Ethics 4 Group Housing 83
3 Why Investigators Should Care about Biomedical 13 Putative Advantages and Disadvantages
Animal Research Ethics 5 of Group Housing 84
4 Aspects of Animal use and Care Relevant to Biomedical 14 Principles of the 3R—Replacement, Reduction,
Animal Research Ethics 8 Refinement 84
5 Use of Privately Owned Animals in Biomedical Research 9 15 Getting the Most Out of an Animal
6 The Nature of Basic Animal Research 10 Experimental Study 85
7 Why Investigators Play the Key Role in Ensuring 16 Need to Correct for Multiple Comparisons? 86
the Ethical Conduct of Animal Research Projects 12 17 Replication Studies 86
8 Sources of Guidance for Investigators in Conducting 18 Identification of Possible Confounds 87
Ethical Research 17 19 Effects of Obesity on Experimental Results 88
9 Developing Useful Ethical Guidelines 19 20 Testing Under Uniform Conditions in the Laboratory
10 Fundamental Principles of Biomedical Animal Versus Testing in a Heterogeneous Environment,
Research Ethics 20 Such as a Farm 89
11 Practical Ethical Guidelines for Investigators 37 21 Training and Testing May Act as Environmental
12 Some Current Difficult Issues in Animal Research Ethics 39 Enrichment 90
13 General Suggestions for Investigators 41 22 Modeling Early Live Events That Affect Subsequent
References 42 Development 90
23 Brain Infarction, Hemorrhage, Traumatic
2. Psychological Environmental Enrichment Brain Injury 91
of Animals in Research 24 Aging and Aging-Related Diseases 91
KRISTINE COLEMAN, JAMES L. WEED, STEVEN J. SCHAPIRO 25 Transgenic Large Animal Models 92
26 Discussion 92
1 Introduction 47 References 95
2 Enrichment and Welfare 48
3 Enrichment and Animal Models 55
4 Enrichment and Experimental Variability 58
5 Implementing an Enrichment Plan 58 Part B
6 Example of an Enrichment Plan: Black-Tailed Prairie
Dogs (Cynomys ludovicianus) 61
VISION
7 Conclusions 62
References 62 4. Animal Models in Cataract Research
JULIE C. LIM, IRENE VORONTSOVA, RENITA M. MARTIS,
3. Large Farm Animal Models of Human PAUL J. DONALDSON
Neurobehavioral and Psychiatric Disorders:
Methodological and Practical Considerations 1 Introduction 103
FRANZ J. VAN DER STAAY, REBECCA E. NORDQUIST, SASKIA S. ARNDT
2 Cataract Types 104
3 Age-Related Nuclear Cataract 105
1 Animal Models 72 4 Diabetic Cortical Cataract 105
2 Why Animal Experimental Studies? 72 5 Animal Models of Diabetic Cataract 109

vii
viii CONTENTS

6 Assessment of Diabetic Cataract Animal Models 112 8 Functional Parameters 194

7 Conclusions 112 References 199
References 113
8. Animal Models of Atherosclerosis
5. N-Methyl-N-Nitrosourea Animal Models GODFREY S. GETZ, CATHERINE A. REARDON
for Retinitis Pigmentosa
AIRO TSUBURA, YUKO EMOTO, MAKI KURO, KATSUHIKO YOSHIZAWA
1 Primate Models 206
2 Porcine Models 207
1 Introduction 118 3 Rabbit Models 208
2 Time-Course Progression of MNU-Induced 4 Mouse Models and Atherosclerosis 209
Retinal Degeneration 119 5 Concluding Comments 215
3 Retinal Degeneration Caused by MNU in References 215
Various Animal Species 120
4 Age-Related Photoreceptor Cell Damage and
Sensitivity to MNU
5 Photoreceptor Cell Death, Cell Debris Removal, and
122 Part D
RPE Cell Migration 123 OBESITY, DIABETES, METABOLIC,
6 Molecular Mechanisms in Photoreceptor Cell Death AND LIVER
Caused by MNU 126
7 Therapeutic Trials Against MNU-Induced
Photoreceptor Apoptosis 129 9. Animal Models of Metabolic Syndrome
8 Concluding Remarks 136 JESSICA P. WAYHART, HEATHER A. LAWSON
9 Appendix: Special Techniques 136
References 139 1 Introduction and Overview 221
2 Choosing an Animal Model of MetS 222
3 Animal Models of MetS Etiology 224
4 Genetic Factors 224
Part C 5 Environmental Factors 232
CARDIAC AND CARDIOVASCULAR 6 Animal Models of MetS Pathophysiology 234
7 Conclusions 237
References 237
6. Animal Models for Cardiovascular Research
YOU-TANG SHEN, LI CHEN, JEFFREY M. TESTANI, 10. Animal Models of Type 1
CHRISTOPHER P. REGAN, RICHARD P. SHANNON and Type 2 Diabetes Mellitus
AILEEN KING, AMAZON AUSTIN
1 Introduction 147
2 Myocardial Ischemic Models 148
1 Introduction 245
3 Hypertension and Left Ventricular Hypertrophy
2 Type 1 Diabetes 246
Models 158
3 Type 2 Diabetes 252
4 Heart Failure Models 161
4 Diabetic Complications 258
5 Models Without Cardiovascular Diseases 168
5 Gender, Strain, and Age Effects in Animal Models
6 Future Directions 172
of Diabetes 259
References 172
6 Conclusions 259
References 259
7. Cardiovascular Models: Heart Secondarily
Affected by Disease (Diabetes Mellitus, 11. Model Nematodes in Obesity Research
Renal Failure, Dysfunctional Sympathetic YU NIE, STEPHEN R. STÜRZENBAUM
Innervation)
JITKA SVIGLEROVA, JITKA KUNCOVA, MILAN STENGL 1 Obesity is a Global Crisis 267
2 Obesity: An Unsolved Medical Problem 268
1 The Heart and Diabetes Mellitus 175 3 Model Organisms Used for Biomedical Research 268
2 Methodological Aspects 176 4 The Nematode Caenorhabditis elegans:
3 Experimental Results in the Rat Model of Small is Beautiful 269
Streptozotocin-Induced Diabetes 179 5 The Basic Biology of Obesity: From a Worm’s
4 Experimental Results in Rat Model of Renal Failure 183 Perspective 271
5 The Heart and Dysfunctional Sympathetic Innervation 190 6 Key Features of Fat Metabolism
6 Methods of Chemical Sympathectomy 191 in Caenorhabditis elegans 271
7 Experimental Results in the Rat Model of Chemical 7 Technical Advances Driving Lipid Research in Nematodes 273
Sympathectomy 191 8 The Intestine: A Driver of Lipid Metabolism 274

  
 CONTENTS ix
9 The Lipid Droplets are the Main Site of Triglyceride 4 Animal Models of Skin Cancer 367
Accumulation 274 5 Conclusions 370
10 The Nile Red Vital Staining Conundrum 275 References 371
11 Lysosome-Related Organelles Participate
in Lipid Mobilization 275
12 Intestinal Autofluorescence: Friend or Foe?
13 A Whole Genome Approach
276
277
Part F
14 New Avenues of Label-Free Methods 277 URINARY TRACT, KIDNEY,
15 Trends and Challenges 277
References 278
AND BOWEL

12. Animal Models for Manipulation 16. Animal Models of Kidney Disease
of Thermogenesis ZAHRAA MOHAMMED-ALI, RACHEL E. CARLISLE, SAMERA NADEMI,
JEFFREY G. DICKHOUT
JOHN-PAUL FULLER-JACKSON, IAIN J. CLARKE,
BELINDA A. HENRY
1 Introduction 379
2 Acute Kidney Disease 379
1 Introduction 281
3 Chronic Kidney Disease: Animal Models 391
2 Thermogenesis—A Significant Determinant
4 Conclusions 406
of Energy Expenditure 282
References 406
3 Concluding Remarks 304
References 304
17. Animal Models to Study Urolithiasis
13. Animal Models of Liver Diseases DAVID T. TZOU, KAZUMI TAGUCHI, THOMAS CHI,
MARSHALL L. STOLLER
YOSHIHISA TAKAHASHI, TOSHIO FUKUSATO

1 Introduction 419
1 Introduction 313
2 Rat Model 420
2 Classical Models of Liver Fibrosis 314
3 Mouse Model 425
3 Animal Models of Specific Liver Diseases 315
4 Fly Model 428
4 Conclusions 333
5 Porcine Model 434
References 333
6 Other Animal Models 436
7 Canine Model 437
8 Feline Model 439
Part E 9 Conclusions 439
References 440
SKIN
18. Animals Models for Healing Studies
14. Animal Models of Skin Regeneration After Partial Nephrectomy
BARBARA GAWRONSKA-KOZAK, JOANNA BUKOWSKA DIOGO B. DE SOUZA, MARCO A. PEREIRA-SAMPAIO,
FRANCISCO J.B. SAMPAIO
1 Introduction 343
2 Skin Healing in Lower Vertebrate (Anamniotes) 1 Introduction 445
Model Organisms 344 2 The Pig as Animal Model 446
3 Skin Healing in Higher Vertebrate (Amniotes) 3 The Sheep as Animal Model 453
Model Organisms 348 4 The Rabbit as Animal Model 459
4 Genetic Mouse Models for Regenerative Skin 5 The Dog as Animal Model 461
Wound Healing (Transcription Factors in Skin 6 Concluding Remarks 462
Development and Regeneration) 350 References 462
5 Conclusions 353
References 353 19. Animal Models of Inflammatory
Bowel Disease
15. Animal Models of Skin Disorders SUMIT JAMWAL, PUNEET KUMAR
JENNIFER Y. ZHANG
1 Introduction 467
1 Introduction 357 2 Historical Perspectives 468
2 Inflammatory Skin Disease 3 Pathophysiology of IBD 468
Animal Models 359 4 Animal Models of Inflammatory Bowel Diseases 469
3 Genetic Skin Disease 5 Conclusions 476
Animal Models 366 References 477

  
x CONTENTS

Part G Part I
STROKE AND NEUROMUSCULAR GENETICS

20. Animal Models of Ischemic Stroke Versus 24. Mouse Models for the Exploration
Clinical Stroke: Comparison of Infarct size, of Klinefelter’s Syndrome
Cause, Location, Study Design, and Efficacy JOACHIM WISTUBA, CRISTIN BRAND, STEFFI WERLER,
LARS LEWEJOHANN, OLIVER S. DAMM
of Experimental Therapies
VICTORIA E. O’COLLINS, GEOFFREY A. DONNAN, 1 Introductory Remarks 621
MALCOLM R. MACLEOD, DAVID W. HOWELLS
2 Klinefelter’s Syndrome—An Underestimated Disease 622
1 Introduction 482 3 The X Chromosome in the Male 623
2 Systematic Review and Metaanalysis Method 482 4 Clinical Features of Klinefelter’s Syndrome 625
3 Results 485 5 Sex Chromosomal Aberrations in Male Mammals 627
4 Discussion 511 6 Mouse Models for Klinefelter’s Syndrome 629
5 Conclusions 517 7 Lessons From Animal Experiments 631
References 518 8 Perspectives—What can be Expected From Future
Animal Experiments and How to Retranslate Experimental
Findings into Clinical Routine—Conclusive Remarks 644
21. The Importance of Olfactory and Motor
References 645
Endpoints for Zebrafish Models
of Neurodegenerative Disease 25. Zebrafish: A Model System to Study the
ANGELA L. SHAMCHUK, W. TED ALLISON, KEITH B. TIERNEY Architecture of Human Genetic Disease
ERICA E. DAVIS, NICHOLAS KATSANIS
1 Introduction 525
2 Building Relevant Models 529
1 Introduction 651
3 Olfactory–Neuromuscular Diseases 532
2 Zebrafish in the Laboratory: A Historical Overview 655
4 Conclusions 544
3 Forward Genetics: Phenotype-Driven Studies
References 545
of Vertebrate Development 656
4 Reverse Genetics: Testing Candidate Genes
in Zebrafish Models 657
Part H 5 Humanizing Zebrafish to Study Human Genetic Variation
6 Modeling Adult Onset Disease in Embryonic or
660

BEHAVIOR Larval Stages 664

7 Therapeutic Discovery in Zebrafish Models of Disease 664
8 Conclusions: The Future of Zebrafish as a
22. Animal Models for Studying Substance Human Genetic Disease Model 665
Use Disorder: Place and Taste Conditioning References 666
CATHERINE M. DAVIS
26. Genetically Tailored Pig Models for
1 What is Substance Use Disorder and Why Should Translational Biomedical Research
We Study It? 557
BERNHARD AIGNER, BARBARA KESSLER, NIKOLAI KLYMIUK, MAYUKO
2 Reward and Reinforcement 558 KUROME, SIMONE RENNER, ANNEGRET WÜNSCH, ECKHARD WOLF
3 Aversive Drug Effects 559
4 The Place Conditioning Procedure 560 1 Introduction 672
5 The Flavor Conditioning Procedure 571 2 Techniques Used for the Generation of Genetically
6 Conclusions 580 Engineered Pigs 673
References 581 3 Genetically Engineered Pigs as Models for Human Diseases 676
4 Conclusions 696
23. Modeling Schizophrenia in Animals References 697
DAVID FEIFEL, PAUL D. SHILLING
27. Genetically Modified Animal Models
1 Overview of Schizophrenia 587 LUCAS M. CHAIBLE, DENISE KINOSHITA, MARCUS A. FINZI CORAT,
2 Approaches to Create Animal Models with Relevance MARIA L. ZAIDAN DAGLI
to Schizophrenia 590
3 Features of Schizophrenia That can be Modeled 1 Introduction 703
in Animals 593 2 Some Historical Aspects 704
4 Specific Animal Models 600 3 Techniques for the Creation of Genetically
References 610 Modified Animals 704

  
 CONTENTS xi
4 Types of Genetically Modified Animals and how 4 Other Animal Models of Early Life Seizures 759
They are Produced 709 5 Mechanisms Underlying Hyperthermia-Induced
5 Genetically Modified Mice as Models of Experimental Febrile Seizures 759
Human Diseases 711 6 Neuroanatomical Changes After Experimental
6 Multifactorial and Polygenic (Complex) Disorders 711 Febrile Seizures 761
7 Inflammatory Diseases 713 7 Neurophysiological Changes After Experimental
8 Neurodegenerative Diseases 713 Febrile Seizures 763
9 Cancer 716 8 Neuronal Hyperactivity After Experimental
References 720 Febrile Seizures 764
9 Behavioral Changes After Experimental
28. Forward and Reverse Genetics to Febrile Seizures 764
Model Human Diseases in the Mouse 10 Conclusions 765
YOICHI GONDO, SHIGERU MAKINO, RYUTARO FUKUMURA
References 765

1 Genetic or Environmental 728 30. Infection-Associated Preterm Birth:

2 Genome Project and Human Diseases 728
Advances From the Use of Animal Models
3 Basic Genetics to Develop and Use Model Mice 729
MATTHEW W. KEMP, GABRIELLE C. MUSK,
4 Diploid and Genotype 730
HARUO USUDA, MASATOSHI SAITO
5 Coisogenic and Congenic Strains 731
6 Double Stranded DNA, Linkage, and Haplotype 732 1 Introduction 769
7 Mutant Mice as Disease Models 732 2 Infection, Inflammation, and Preterm Birth 771
8 Fancy Mice 733 3 Innate Immune Responses 774
9 Laboratory Mouse Strains 733 4 Inflammation and Labor 776
10 Redundancy of Genes: Oculocutaneous Albinism 733 5 The Use of Animals in the Study of Preterm Birth—
11 Body Weight and Brain Function: Pleiotropy Justification and Validity 777
of ob and db 734 6 Animal Models of Infection-Associated Inflammation 778
12 Conventional Positional Cloning and Forward 7 Summary 784
Genetics 734 8 Practical Study—Fetal Surgery in the Sheep 784
13 Unique Positional Cloning: High Reversion Rates of dv References 798
and pun Mutations 735
14 Recombinant Inbred Strains for Quick
Genetic Mapping 735 31. Animal Models for the Study
15 Mutagenesis for Forward Genetics 736 of Neonatal Disease
16 Large-Scale ENU Mouse Mutagenesis Project 738 JEAN-PAUL PRAUD, YUICHIRO MIURA, MARTIN G. FRASCH
17 Mutagenesis for Reverse Genetics 740
18 Gene Targeting and Knockout Mouse 740 Neonatal Respiration 806
19 Transgenic Mice as Disease Models 740 1 Establishment of Air Breathing at Birth 806
20 Knockout Mice as Disease Models 741 2 Neonatal Lung Diseases 811
21 Conditional Targeting 741 Patent Ductus Arteriosus 814
22 International Knockout Mouse Consortium 741 1 Introduction 814
23 ENU-Based Reverse Genetics in the Mouse 742 2 Normal Physiological Course of Ductus
24 Further Advancement of Genome Technologies 743 Arteriosus 815
25 Genome Editing Technologies 744 3 Pathophysiology of Patent Ductus Arteriosus 815
26 Concluding Remarks 746 4 Treatment for Patent Ductus Arteriosus 815
References 747 5 Animal Models of Patent Ductus Arteriosus 815
Retinopathy of Prematurity 816
1 Introduction 816

Part J 2 Normal Vascularization of Human Retina

3 Pathophysiology of Retinopathy of Prematurity
816
817
EARLY LIFE 4 Treatment of Retinopathy of Prematurity 817
5 Animal Models of Retinopathy of Prematurity 817
Intraventricular Hemorrhage 819
29. Experimental Febrile Seizures in Rodents 1 Introduction 819
RYUTA KOYAMA 2 Pathophysiology of Intraventricular Hemorrhage 819
3 Prevention of Intraventricular Hemorrhage 820
1 Introduction 755 4 Animal Models of Intraventricular Hemorrhage 820
2 Febrile Seizures in Humans and Their Relationship Cerebral Palsy 820
to Epilepsy 756 1 Epidemiology, Etiology, and Animal Models 821
3 Animal Models of Febrile Seizures (Experimental 2 Fetal Inflammation 821
Febrile Seizures) 757 3 Fetal Acidemia 822

  
xii CONTENTS

Necrotizing Enterocolitis
1 Etiology
823
823
Part L
2 Chorioamnionitis: Pathological Fetal Inflammation CANCER
is a Risk Factor 823
3 Tight Junctions: Intestinal Permeability And Integrity 824
4 Intestinal Permeability is Influenced by Pathogens 34. Modeling Cancer Using
and Inflammation 824 CRISPR-Cas9 Technology
5 ENS Controls Epithelial Barrier Function 824 SANDRA RODRIGUEZ-PERALES, MARTA MARTINEZ-LAGE,
6 NEC: Immature Immune Response 825 RAUL TORRES-RUIZ
7 Development and Monitoring of the Autonomic Nervous
System Activity 825 1 Introduction 905
8 CAP Controls Immune Homeostasis 825 2 Generation of CRISPR Cancer Models 911
9 Vagal Nerve Stimulation and the Gut Inflammation 826 3 Challenges and Solutions 917
10 Near Future: Early Detection of NEC Using Fetal 4 Concluding Remarks 919
and Neonatal Heart Rate Monitoring? 827 Glossary 919
References 827 References 920

32. Animal Models of Fetal Programming: 35. Modeling Breast Cancer in Animals—
Focus on Chronic Maternal Stress During Considerations for Prevention and Treatment Studies
Pregnancy and Neurodevelopment JOELLEN WELSH

MARTIN G. FRASCH, JAY SCHULKIN, GERLINDE A.S. METZ,

MARTA ANTONELLI
1 Introduction to Animal Models of Breast Cancer 926
2 Concepts of Breast Cancer Biology 926
1 Introduction 839 3 Modeling Breast Cancer in Rodents 928
2 Significance 841 4 Spontaneous and Induced Mammary Tumorigenesis
3 Preclinical Studies of Prenatal Stress in Rodent Models 842 in Rodents 928
4 Experimental Paradigm for a Trans- and 5 Grafting and Transplantation Approaches 932
Multigenerational PS Rat Model 845 6 Genetically Engineered Mouse Models
5 Large Animal Models of Fetal Development of Breast Cancer 940
to Model PS: Pregnant Sheep 845 7 Comparative Pathology and Genomics: Mouse
6 Experimental Paradigm for Prenatal Stress Model Mammary Tumors Versus Human Breast Cancer 942
in Fetal Sheep 846 8 Studying Metastasis in Mice With an Emphasis
7 Conclusions 846 on New Models 943
References 846 9 Emerging Nonrodent Models of Breast Cancer 944
10 Conclusions 944
References 945

Part K
VIRAL DISEASE Part M
SCLEROSIS
33. Animal Models of Human Viral Diseases
SARA I. RUIZ, ELIZABETH E. ZUMBRUN, AYSEGUL NALCA
36. Animal Models of Systemic Sclerosis
1 Introduction 853 TOSHIYUKI YAMAMOTO
2 Caliciviridae 854
3 Togaviridae 855 1 Introduction 951
4 Flaviviridae 858 2 Bleomycin-Induced Murine Scleroderma 952
5 Coronaviridae 861 3 HOCl-Induced Murine Scleroderma 957
6 Filoviridae 864 4 Tight Skin Mouse 957
7 Orthomyxoviridae 870 5 Sclerodermatous Graft-Versus-Host
8 Bunyaviridae 873 Disease Model 959
9 Arenaviridae 876 6 Skin Fibrosis by Exogenous Injection
10 Retroviridae 877 of Growth Factors 959
11 Papillomaviridae 879 7 UCD-200 Chicken 960
12 Poxviridae 880 8 Transgenic Mouse Models 960
13 Hepadnaviridae 883 9 Knockout Mouse Models 960
14 Conclusions 884 10 Conclusions 961
References 885 References 961

  
 CONTENTS xiii

37. Animal Models for the Study of Multiple Sclerosis 41. Neurotoxin 1-Methyl-4-Phenyl-1,2,3,6-
ROBERT H. MILLER, SHARYL FYFFE-MARICICH, Tetrahydropyridine (MPTP)-Induced Animal
ANDREW C. CAPRARIELLO
Models of Parkinson’s Disease
1 The Complex Biology of Multiple Sclerosis 967 JIRO KASAHARA, MOHAMMED E. CHOUDHURY, NORIKO NISHIKAWA,
AKIE TANABE, RYOSUKE TSUJI, YU ZHOU, MASATOSHI OGAWA,
2 Immunological Models for CNS Demyelination 971 HIRONORI YOKOYAMA, JUNYA TANAKA, MASAHIRO NOMOTO
3 Local Induction of Demyelination Following Injection
of Myelin Peptides 973 1 Introduction 1088
4 Development of MS Therapies Based on Models 2 Clinical Characteristics of PD and Their Relevant
of Immune Mediated Demyelinating Diseases 974 Symptoms in Animal Models 1091
5 Viral mediated Models of Demyelination 974 3 Molecular Pathophysiology of PD 1092
6 Oligodendrocyte Induced Cell Death Models of 4 Neurotoxins for Making PD Models 1093
Demyelination 975 5 MPTP-Induced Mice Model of PD 1095
7 Toxin Models of Demyelination 979 6 MPTP-Induced Common Marmoset Model for PD 1101
8 Conclusions and Comments 984 7 Concluding Remarks 1104
References 984 References 1104

42. Animal Models for the Study of Human

Neurodegenerative Diseases
Part N GABRIELA D. COLPO, FABIOLA M. RIBEIRO, NATALIA P. ROCHA,
PSYCHIATRIC AND ANTÔNIO L. TEIXEIRA

NEUROLOGICAL DISEASE 1 Introduction 1109

2 Parkinson’s Disease 1110
3 Alzheimer’s Disease 1113
38. Animal Models of Mood Disorders: Focus on 4 Huntington’s Disease 1115
Bipolar Disorder and Depression 5 Amyotrophic Lateral Sclerosis 1117
SAMIRA S. VALVASSORI, ROGER B. VARELA, JOÃO QUEVEDO 6 Frontotemporal Dementia 1119
7 Conclusions 1121
1 Introduction 991 References 1124
2 Animal Models of Major Depression 992
3 Animal Models of Mania 997 43. Animal Models of Mania: Essential
4 Conclusions 1000 Tools to Better Understand Bipolar Disorder
References 1000
ALINE S. DE MIRANDA, ROBERTO ANDREATINI,
ANTÔNIO L. TEIXEIRA
39. Pigs as Model Species to Investigate
Effects of Early Life Events on Later Behavioral 1 Introduction 1131
and Neurological Functions 2 Insights From Pharmacological Animal Models of Mania 1132
3 Insights From Environmental Models 1137
REBECCA E. NORDQUIST, ELLEN MEIJER,
FRANZ J. VAN DER STAAY, SASKIA S. ARNDT 4 Insights From Genetic Models 1138
5 Conclusions 1138
1 Introduction 1003 References 1139
2 Operant Tasks for Testing Cognitive Functioning 1004
3 Nonoperant Behavioral Tests 1011 44. Stress Coping and Resilience Modeled in Mice
4 Welfare Aspects in the Use of Pigs as Model Species 1019 DAVID M. LYONS, LUIS DE LECEA, ALAN F. SCHATZBERG
5 Conclusions 1024
References 1024 1 Introduction 1145
2 Learning to Cope Training 1146
40. Animal Models of Alzheimer’s Disease 3 Learning to Cope Is Stressful 1146
4 Learning to Cope Reduces Subsequent Behavioral
MORGAN NEWMAN, DORIS KRETZSCHMAR, IMRAN KHAN,
MENGQI CHEN, GIUSEPPE VERDILE, MICHAEL LARDELLI Measures of Emotionality 1147
5 Learning to Cope Increases Anterior Cingulate
1 Introduction 1031 Cortex Stargazin Gene Expression 1148
2 Invertebrate Models of Alzheimer’s Disease 1032 6 Discussion 1149
3 Nonmammalian Vertebrate Models of Alzheimer’s 7 Limitations 1151
Disease 1041 8 Conclusions 1151
4 Mammalian Models of Alzheimer’s Disease 1051 References 1151
5 Conclusions 1066
References 1066 Index 1155

  
Page left intentionally blank
 List of Contributors
Bernhard Aigner Molecular Animal Breeding and
Biotechnology; Center for Innovative Medical Models
(CiMM), Ludwig-Maximilian University, Munich, Germany
W. Ted Allison University of Alberta, Edmonton, AB,
Canada
Roberto Andreatini Federal University of Paraná, Centro
Politécnico, Curitiba, PR, Brazil
Marta Antonelli Institute of Cellular Biology and
Neuroscience “Prof Dr. Eduardo de Robertis”, Buenos
Aires University, Buenos Aires, Argentina
Saskia S. Arndt Utrecht University, Utrecht, The Netherlands
Amazon Austin King’s College London, London, United
Kingdom
Cristin Brand Institute of Reproductive and Regenerative
Biology, Centre of Reproductive Medicine and Andrology,
University Clinics, Münster, Germany
Joanna Bukowska Institute of Animal Reproduction and
Food Research, Polish Academy of Sciences, Olsztyn, Poland
Andrew C. Caprariello University of Calgary, Hotchkiss
Brain Institute, Canada
Rachel E. Carlisle McMaster University and St. Joseph’s
Healthcare Hamilton, Hamilton, ON, Canada
Lucas M. Chaible Meyerhofstr, Heidelberg, Germany
Li Chen Merck Research Laboratories, Merck & Co., Inc.,
West Point, PA, United States
Mengqi Chen Centre of Excellence for Alzheimer’s Disease
Research and Care, Edith Cowan University, Joondalup, WA,
Australia
Thomas Chi University of California, San Francisco, CA,
United States
Mohammed E. Choudhury Ehime University Graduate
School of Medicine, Shitsukawa, Shitsukawa, Toon-shi,
Ehime, Japan
Iain J. Clarke Monash Biomedical Discovery Institute,
Monash University, Clayton, VIC, Australia
Kristine Coleman Oregon National Primate Research
Center, Beaverton, OR, United States
Gabriela D. Colpo University of Texas Health Science
Center at Houston, Houston, TX, United States
Oliver S. Damm Institute of Reproductive and Regenerative
Biology, Centre of Reproductive Medicine and Andrology,
University Clinics, Münster, Germany
Catherine M. Davis Johns Hopkins University School of
Medicine, Baltimore, MD, United States
Erica E. Davis Center for Human Disease Modeling, Duke
University Medical Center, Durham, NC, United States
xv
Luis de Lecea Stanford University, Stanford, CA, United
States
Aline S. de Miranda Biological Institute, Federal University
of Minas Gerais (UFMG), Belo Horizonte, MG, Brazil
Diogo B. de Souza State University of Rio de Janeiro, Rio de
Janeiro, RJ, Brazil
Jeffrey G. Dickhout McMaster University and St. Joseph’s
Healthcare Hamilton, Hamilton, ON, Canada
Paul J. Donaldson NZ National Eye Centre; University of
Auckland, Auckland, New Zealand
Geoffrey A. Donnan Florey Neuroscience Institutes,
Melbourne Brain Centre, University of Melbourne,
Heidelberg, VIC, Australia
Yuko Emoto Kansai Medical University, Hirakata, Osaka,
Japan
David Feifel University of California, San Diego, La Jolla,
CA, United States
Marcus A. Finzi Corat University of Campinas, Campinas,
São Paulo, Brazil
Martin G. Frasch University of Washington, Seattle, WA,
United States
Ryutaro Fukumura RIKEN BioResource Center, Tsukuba,
Ibaraki, Japan
Toshio Fukusato Teikyo University School of Medicine,
Tokyo, Japan
John-Paul Fuller-Jackson Monash Biomedical Discovery
Institute, Monash University, Clayton, VIC, Australia
Sharyl Fyffe-Maricich University of Pittsburgh, Rangos
Research Center, Pittsburgh, PA, United States
Barbara Gawronska-Kozak Institute of Animal
Reproduction and Food Research, Polish Academy of
Sciences, Olsztyn, Poland
Godfrey S. Getz Ben May Institute for Cancer Biology,
University of Chicago, Chicago, IL, United States
Yoichi Gondo RIKEN BioResource Center, Tsukuba, Ibaraki,
Japan
Belinda A. Henry Monash Biomedical Discovery Institute,
Monash University, Clayton, VIC, Australia
David W. Howells Florey Neuroscience Institutes,
Melbourne Brain Centre, University of Melbourne, Heidelberg,
VIC; University of Tasmania, Hobart, TAS, Australia
Sumit Jamwal I.S.F College of Pharmacy, Moga, Punjab,
India
Jiro Kasahara Institute of Biomedical Sciences, Graduate
School of Pharmaceutical Sciences, Tokushima University,
Shoumachi, Tokushima, Japan
xvi LIST OF CONTRIBUTORS
Nicholas Katsanis Center for Human Disease Modeling,
Duke University Medical Center, Durham, NC, United States
Matthew W. Kemp The University of Western Australia,
Perth, Australia; Tohoku University Hospital, Sendai, Japan
Barbara Kessler Molecular Animal Breeding and
Biotechnology; Center for Innovative Medical Models
(CiMM), Ludwig-Maximilian University, Munich, Germany
Imran Khan Curtin Health Innovation Research Institute—
Biosciences, Curtin University, Bentley; The University of
Western Australia, Crawley; Hollywood Medical Centre,
Nedlands, WA, Australia
Aileen King King’s College London, London, United
Kingdom
Denise Kinoshita Ibirapuera University, Interlagos,
São Paulo, Brazil
Nikolai Klymiuk Molecular Animal Breeding and
Biotechnology; Center for Innovative Medical Models
(CiMM), Ludwig-Maximilian University, Munich, Germany
Ryuta Koyama Graduate School of Pharmaceutical Sciences,
The University of Tokyo, Tokyo, Japan
Doris Kretzschmar Oregon Institute of Occupational Health
Sciences, Oregon Health & Sciences University, Portland, OR,
United States
Puneet Kumar I.S.F College of Pharmacy, Moga, Punjab,
India
Jitka Kuncova Biomedical Center, Charles University,
Pilsen, Czech Republic
Maki Kuro Kansai Medical University, Hirakata, Osaka,
Japan
Mayuko Kurome Molecular Animal Breeding and
Biotechnology; Center for Innovative Medical Models
(CiMM), Ludwig-Maximilian University, Munich, Germany
Michael Lardelli University of Adelaide, Adelaide,
SA, Australia
Heather A. Lawson Washington University School of
Medicine, St. Louis, MO, United States
Lars Lewejohann University of Münster, Münster,
Germany
Julie C. Lim NZ National Eye Centre; University of
Auckland, Auckland, New Zealand
David M. Lyons Stanford University, Stanford, CA, United
States
Malcolm R. Macleod University of Edinburgh, Edinburgh,
United Kingdom
Shigeru Makino RIKEN BioResource Center, Tsukuba,
Ibaraki, Japan
Marta Martinez-Lage Spanish National Cancer Research
Centre—CNIO, Madrid, Spain
Renita M. Martis NZ National Eye Centre; University of
Auckland, Auckland, New Zealand
Ellen Meijer Utrecht University, Utrecht, The Netherlands
Gerlinde A.S. Metz Canadian Centre for Behavioural
Neuroscience, University of Lethbridge, Lethbridge,
Alberta, Canada
  
Robert H. Miller George Washington University,
Washington, DC, United States
Yuichiro Miura The University of Western Australia, Perth,
Australia
Zahraa Mohammed-Ali McMaster University and St.
Joseph’s Healthcare Hamilton, Hamilton, ON, Canada
Gabrielle C. Musk Animal Care Services, The University of
Western Australia, Perth, Australia
Samera Nademi McMaster University and St. Joseph’s
Healthcare Hamilton, Hamilton, ON, Canada
Aysegul Nalca Center for Aerobiological Sciences, United
States Army Institute of Infectious Diseases (USAMRIID), Fort
Detrick, Frederick, MD, United States
Morgan Newman University of Adelaide, Adelaide, SA,
Australia
Yu Nie King’s College London, London, United Kingdom
Noriko Nishikawa Ehime University Graduate School
of Medicine, Shitsukawa, Shitsukawa, Toon-shi, Ehime,
Japan
Masahiro Nomoto Ehime University Graduate School of
Medicine, Shitsukawa, Shitsukawa, Toon-shi, Ehime, Japan
Rebecca E. Nordquist Brain Center Rudolf Magnus; Utrecht
University, Utrecht, The Netherlands
Victoria E. O’Collins Florey Neuroscience Institutes,
Melbourne Brain Centre, University of Melbourne,
Heidelberg, VIC, Australia
Masatoshi Ogawa Institute of Biomedical Sciences,
Graduate School of Pharmaceutical Sciences, Tokushima
University, Shoumachi, Tokushima, Japan
Marco A. Pereira-Sampaio State University of Rio de
Janeiro, Rio de Janeiro; Fluminense Federal University,
Niteroi, RJ, Brazil
Jean-Paul Praud Université de Sherbrooke, Sherbrooke, QC,
Canada
João Quevedo University of Southern Santa Catarina
(UNESC), Criciúma, Santa Catarina, Brazil; Center for
Experimental Models in Psychiatry, The University of Texas
Medical School at Houston, Houston, TX, United States
Catherine A. Reardon Ben May Institute for Cancer Biology,
University of Chicago, Chicago, IL, United States
Christopher P. Regan Merck Research Laboratories, Merck
& Co., Inc., West Point, PA, United States
Simone Renner Molecular Animal Breeding and
Biotechnology; Center for Innovative Medical Models
(CiMM), Ludwig-Maximilian University, Munich, Germany
Fabiola M. Ribeiro Institute of Biological Sciences, Federal
University of Minas Gerais, (UFMG), Belo Horizonte, MG, Brazil
Natalia P. Rocha University of Texas Health Science Center
at Houston, Houston, TX, United States
Sandra Rodriguez-Perales Spanish National Cancer
Research Centre—CNIO, Madrid, Spain
Sara I. Ruiz Center for Aerobiological Sciences, United
States Army Institute of Infectious Diseases (USAMRIID),
Fort Detrick, Frederick, MD, United States
 LIST OF CONTRIBUTORS xvii
Masatoshi Saito The University of Western Australia, David T. Tzou University of California, San Francisco, CA,
Perth, Australia; Tohoku University Hospital, Sendai, United States
Japan Haruo Usuda The University of Western Australia,
Francisco J.B. Sampaio State University of Rio de Janeiro, Perth, Australia; Tohoku University Hospital, Sendai, Japan
Rio de Janeiro, RJ, Brazil Samira S. Valvassori University of Southern Santa Catarina
Steven J. Schapiro The University of Texas MD Anderson (UNESC), Criciúma, Santa Catarina, Brazil
Cancer Center, TX, United States; The University of Franz J. van der Staay Brain Center Rudolf Magnus; Utrecht
Copenhagen, København, Denmark University, Utrecht, The Netherlands
Alan F. Schatzberg Stanford University, Stanford, CA, Roger B. Varela University of Southern Santa Catarina
United States (UNESC), Criciúma, Santa Catarina, Brazil
Jay Schulkin University of Washington, Seattle, WA, Giuseppe Verdile Curtin Health Innovation
United States Research Institute—Biosciences, Curtin University,
Angela L. Shamchuk University of Alberta, Edmonton, AB, Bentley; The University of Western Australia, Crawley;
Canada Hollywood Medical Centre, Nedlands, WA, Australia
Richard P. Shannon University of Virginia, Charlottesville, Irene Vorontsova University of California, Irvine, Irvine,
VA, United States CA, United States
You-Tang Shen University of Pennsylvania, Philadelphia, Jessica P. Wayhart Washington University School of
PA, United States Medicine, St. Louis, MO, United States
Paul D. Shilling University of California, San Diego, La James L. Weed Comparative Medicine Branch, National
Jolla, CA, United States Center for Emerging and Zoonotic Infectious Diseases,
Milan Stengl Biomedical Center, Charles University, Pilsen, Centers for Disease Control and Prevention, Atlanta, GA,
Czech Republic United States
Marshall L. Stoller University of California, San Francisco, JoEllen Welsh Cancer Research Center, University at
CA, United States Albany, Albany, NY, United States
Stephen R. Stürzenbaum King’s College London, London, Steffi Werler Institute of Reproductive and Regenerative
United Kingdom Biology, Centre of Reproductive Medicine and Andrology,
University Clinics, Münster, Germany
Jitka Sviglerova Biomedical Center, Charles University,
Pilsen, Czech Republic Joachim Wistuba Institute of Reproductive and
Regenerative Biology, Centre of Reproductive Medicine and
Kazumi Taguchi University of California, San Francisco,
Andrology, University Clinics, Münster, Germany
CA, United States; Nagoya City University Graduate School
of Medical Sciences, Nagoya, Japan Eckhard Wolf Molecular Animal Breeding and
Biotechnology; Center for Innovative Medical Models
Yoshihisa Takahashi Teikyo University School of Medicine,
(CiMM); Gene Center, Ludwig-Maximilian University,
Tokyo, Japan
Munich, Germany
Akie Tanabe Institute of Biomedical Sciences, Graduate
Annegret Wünsch Molecular Animal Breeding and
School of Pharmaceutical Sciences, Tokushima University,
Biotechnology; Center for Innovative Medical Models
Shoumachi, Tokushima, Japan
(CiMM), Ludwig-Maximilian University, Munich, Germany
Junya Tanaka Ehime University Graduate School of
Toshiyuki Yamamoto Fukushima Medical University,
Medicine, Shitsukawa, Shitsukawa, Toon-shi, Ehime,
Fukushima, Japan
Japan
Hironori Yokoyama Institute of Biomedical Sciences,
Jerrold Tannenbaum University of California, Davis, Davis,
Graduate School of Pharmaceutical Sciences, Tokushima
CA, United States
University, Shoumachi, Tokushima, Japan
Antônio L. Teixeira University of Texas Health Science
Katsuhiko Yoshizawa Kansai Medical University, Hirakata,
Center at Houston, Houston, TX, United States
Osaka; Mukogawa Women’s University, Nishinomiya,
Jeffrey M. Testani Yale University, New Haven, CT, United Hyogo, Japan
States
Maria L. Zaidan Dagli School of Veterinary Medicine and
Keith B. Tierney University of Alberta, Edmonton, AB, Animal Science, University of São Paulo, São Paulo, Brazil
Canada
Jennifer Y. Zhang Duke University Medical Center,
Raul Torres-Ruiz Spanish National Cancer Research Durham, NC, United States
Centre—CNIO, Madrid, Spain
Yu Zhou Institute of Biomedical Sciences, Graduate
Airo Tsubura Kansai Medical University, Hirakata, Osaka, School of Pharmaceutical Sciences, Tokushima University,
Japan Shoumachi, Tokushima, Japan
Ryosuke Tsuji Institute of Biomedical Sciences, Graduate Elizabeth E. Zumbrun United States Army Institute of
School of Pharmaceutical Sciences, Tokushima University, Infectious Diseases (USAMRIID), Fort Detrick, Frederick, MD,
Shoumachi, Tokushima, Japan United States

  
Page left intentionally blank
 Preface
The key to use animal models is to select ones that
have similarities to human tissues or disease states.
Some models have the advantage that the reproductive,
mitotic, development, or aging cycles are more rapid
compared with those in humans, facilitating study; oth-
ers are utilized because individual proteins may be stud-
ied in an advantageous way and have human homologs.
Other organisms are facile to grow in laboratory settings,
lend themselves to convenient analyses or have defined
properties that are important.
The collection of systems represented in this volume
is an effort to reflect the diversity and utility of models
that are used to study human disease. That utility is
based on the consideration that observations made in
particular organisms provide insight into the workings
xix
of other, more complex systems. Even the cell cycle in the
simple yeast cell has similarities to that in humans and
regulation with similar proteins occurs.
We have included sections on ethics and animal care,
and the ethics of research; both are important in our view.
Some models will have been omitted due to page
limitations and we have encouraged the authors to use
tables and figures to make comparisons of models so
that observations not available in primary publications
can become useful to the reader.
We thank the authors for the timely submission of
chapters so that this volume could be published while it
is still fresh and to the staff at Academic Press for shep-
herding the process of publication.
P. Michael Conn
Lubbock, TX, United States
Page left intentionally blank
 P A R T A

ETHICS, RESOURCES,
AND APPROACHES
1 Ethics in Biomedical Animal Research: The Key Role
of the Investigator 3
2 Psychological Environmental Enrichment of Animals
in Research 47
3 Large Farm Animal Models of Human Neurobehavioral
and Psychiatric Disorders: Methodological and Practical
Considerations 71
Page left intentionally blank
 C H A P T E R

1
Ethics in Biomedical Animal Research:
The Key Role of the Investigatora
Jerrold Tannenbaum
University of California Davis, Davis, CA, United States

O U T L I N E

1 Nature and Scope of the Chapter 4 6.3 What Investigators Mean by “Basic”
Animal Research 11
2 The Subject Matter of Biomedical
6.4 Recommendations 11
Animal Research Ethics 4
7 Why Investigators Play the Key Role
3 Why Investigators Should Care About
in Ensuring the Ethical Conduct
Biomedical Animal Research Ethics 5
of Animal Research Projects 12
3.1 Investigators are Already Committed
7.1 The Nature of Principles of Biomedical
to High Ethical Standards 5
Animal Research Ethics 12
3.2 Ethical Treatment of Animals
7.2 The Need for Scientific Expertise
is Required by Law 5
in Applying General Ethical Principles 12
3.3 Ethical Treatment of Animals Promotes
7.3 The Investigator as Captain
Sound Scientific Results 6
of the Experimental Ship 12
3.4 Ethical Treatment of Animals is Required
7.4 The Role of Review Committees
by Funding Bodies 6
and Government Officials in Assessing
3.5 Ethical Treatment of Animals is Required
the Ethical Appropriateness of Animal
by Peer-reviewed Journals 7
Experiments 14
3.6 Ethical Treatment of Animals is Essential
for Public Support 8 8 Sources of Guidance for Investigators
3.7 Ethical Treatment of Animals in Conducting Ethical Research 17
is the Right Thing to Do 8 8.1 Scientific Resources 17
4 Aspects of Animal Use and Care Relevant 8.2 Ethical and Ethically Relevant Resources 17
to Biomedical Animal Research Ethics 8 9 Developing Useful Ethical Guidelines 19
5 Use of Privately Owned Animals 10 Fundamental Principles of Biomedical Animal
in Biomedical Research 9 Research Ethics 20
6 The Nature of Basic Animal Research 10 F1 The Biomedical Research Principle 20
6.1 “Basic” Research as the Search for Knowledge F2 The Animal Research Principle 21
for its Own Sake 10 F3 The Nuremberg Principle 21
6.2 “Basic” Research as the Search for Fundamental, F4 The Pain and Distress Minimization
Underlying Mechanisms, and Causes 11 Principle 22

a
Some content in this chapter has been reproduced from Tannenbaum, J., 2017. Ethics in the Use of Animal Models of Seizures and Epilepsy. In:
Pitkänen, A., et al. (Eds). Models of Seizures and Epilepsy, Chapter 5, pp. 47-68.
Animal Models for the Study of Human Disease. 
Copyright © 2017 Elsevier Inc. All rights reserved.
3
4 1. Ethics in Biomedical Animal Research: The Key Role of the Investigatora
F5 The Pain and Distress Justification Principle 23
F6 The Harm Justification Principle 24
F7 The Harm Minimization Principle 30
F8 The General Justification Principle 30
F9 The 3Rs Principle 31
F10 The Species-Appropriate Housing
Principle 33
F11 The Appropriate Care Principle 35
F12 The Appreciation Principle 36
F13 The Translation Principle 36
11 Practical Ethical Guidelines
for Investigators 37
1 NATURE AND SCOPE
OF THE CHAPTER
This chapter presents an overview and explanation
of ethical principles that are of crucial importance in the
use of animal models for the study of human disease.
The chapter is intended primarily for, and is written
from the perspective of, scientists who conduct biomedi-
cal animal research. Like the other chapters in this vol-
ume, the discussion of animal research ethics presented
here focuses on matters that are directly relevant to the
design and implementation, by investigators, of animal
research projects.
The chapter begins with a consideration of the nature
of biomedical animal research ethics. The chapter reviews
the reasons investigators should engage in ethical assess-
ment of their work, and explains why they are uniquely
qualified to do so. The discussion presents a number of
fundamental principles of biomedical animal research eth-
ics, and offers guidelines for putting these principles into
practice. The chapter then highlights several ethical issues
regarding which members of the biomedical research com-
munity have exhibited some lack of clarity or disagree-
ment. The discussion concludes with recommendations
for participation by investigators in ethical discussion of
their work, and of biomedical animal research generally.
The chapter does not provide a detailed description of
ethical theories that have been applied to biomedical ani-
mal research by academic philosophers or legal theorists.
Nor does the chapter summarize the large (and growing)
scholarly literature in animal research ethics. I suggest be-
low that animal researchers become familiar with some of
this literature. However, attempting to present compre-
hensive descriptions of ethical theories or the animal re-
search ethics literature would preclude a discussion that
is useful to investigators. Aside from the fact that many
different arguments and conclusions have been offered
by different thinkers, much of the animal research ethics
A. Ethics, Resources, and Approaches
12 Some Current Difficult Issues in
Animal Research Ethics 39
12.1 The Nature and Weight of Harms
to be Justified by the Value of Animal
Research Project 39
12.2 Ethical Relevance of Species and Species
Characteristics 40
13 General Suggestions for Investigators 41
S1 Communicate to the Public 41
S2 Become Familiar with the Literature 41
S3 Participate in Discussions of Ethical Issues
Relating to Animal Research 42
References 42
literature focuses on a question that has already been an-
swered by scientists who conduct biomedical animal re-
search, and by society generally. As stated by one philo-
sophical discussion, “at the heart of the wide-ranging and
seemingly unending controversy over the use of animals
in biomedical science, whether in basic or applied re-
search, toxicity testing, drug production, or education, is
one burning question: Are we humans ethically justified in
such a use of animals, in general and in particular cases?”
(Donnelly and Nolan, 1990, p. 8) Clearly, there would be
no need for this chapter, or more importantly for biomed-
ical animal research ethics, if there were no significant
ethical issues relating to the use of animals in biomedical
research. However, it is simply a fact that among the most
fundamental ethical tenets of contemporary society is not
just that humans are justified in using animals to under-
stand and combat human disease, but that we are often
ethically obligated to do so.
As used in the chapter, the term “investigator” refers
to principal and coprincipal investigators, and also in-
cludes scientific colleagues who participate significantly
in formulating the goals and design of an animal re-
search project. The central theme of the chapter is that
investigators—not animal care and use committees, at-
tending veterinarians, or government agencies—play
the primary role in ensuring the ethical conduct of bio-
medical animal research.
2 THE SUBJECT MATTER
OF BIOMEDICAL ANIMAL
RESEARCH ETHICS
Biomedical animal research ethics has three essential
features in addition to its concern with the use of ani-
mals in scientific research.
First, biomedical animal research ethics involves issues
that pertain to whether animals employed in research are
 3 Why Investigators Should Care About Biomedical Animal Research Ethics
used and treated properly from an ethical standpoint in
ways that accord with human ethical obligations to the an-
imals. Many important ethical questions raised by uses of
animals in biomedical research are not issues of biomedi-
cal animal research ethics. For example, potential employ-
ment of the CRISPR gene editing technique to produce
animals with transplantable human organs has raised
questions about whether the technique itself could lead to
the editing of the human germ line and to the engineering
of humans with supposedly desirable characteristics (Bal-
timore et al., 2015; Lanphier et al., 2015), or could result in
unintended organisms that might harm public health or
the environment (Caplan et al., 2015). These are not ques-
tions of biomedical animal research ethics because they
do not ask whether the animals in such research are being
used properly, from the standpoint of the interests of the
animals themselves. In contrast, it would be a matter of
biomedical animal research ethics to ask whether animals
produced using CRISPR will experience problematic pain
or distress: during research procedures, as a result of their
genetic make up, or because of how they might be bred
and raised to provide human organs or tissues (Combes
and Balls, 2014; Hyun et al., 2007).
Second, biomedical animal research ethics addresses
questions that pertain to animals employed in research
intended to address medical issues in humans or animals.
Biomedical animal research ethics is a subset or branch of
animal research ethics, which deals with ethical obliga-
tions to animals used in all kinds of scientific animal re-
search, including, for example, use of animals to test the
safety of household products, chemicals used in industrial
processes, and cosmetics; research intended to understand
how various species can be preserved in the wild; and ag-
ricultural experiments aimed at breeding animals that pro-
duce nutritious and palatable food. These uses of animals
sometimes involve facts, ethical issues, and ethical princi-
ples that are significantly different from those that pertain
to using animal models to study human disease. Biomedi-
cal animal research ethics considers research directed at
improving animal health because animals are sometimes
intended beneficiaries of animal research (Quimby, 1998).
Third, as generally defined, biomedical animal re-
search ethics (like animal research ethics) relates to the use
and treatment of live animals that are sentient, in the sense
of being capable of experiencing unpleasant or pleasant
sensations or feelings. Questions in biomedical animal
research ethics can be asked regarding whether live ani-
mals should be killed. For example, some people believe
that certain species (e.g., chimpanzees) should never be
killed in research, and it can be asked whether it is mor-
ally wrong (and not just a waste of resources) to euthanize
many more mice for their tissues than is required for a
given study. These are questions in biomedical animal re-
search ethics because they ask whether there is an ethical
obligation to certain animals to allow them to live.
A. Ethics, Resources, and Approaches
5
This chapter considers issues in biomedical a­ nimal
research ethics relating to the proper treatment of
­animals employed in scientific research that aims at dis-
covering methods of preventing, alleviating, or curing
human disease.
3 WHY INVESTIGATORS SHOULD
CARE ABOUT BIOMEDICAL ANIMAL
RESEARCH ETHICS
Some investigators may be suspicious of advice
(sometimes offered by nonscientists) that they should
devote valuable time and energy to ethical assessment
of their research. Biomedical research is science, and sci-
ence deals with objectively verifiable facts and hypoth-
eses. In contrast, ethics is a matter of value judgments,
about which there can be disagreement, even when
there is an agreement about the facts. For example, some
people believe that it is unethical to use animals in re-
search, even when such research results in the preven-
tion or cure of serious disease (Regan, 2004). It is diffi-
cult enough to conduct productive scientific research,
the results of which are subject to objective empirical
verification. Leave us alone to do our experiments, some
investigators might say, and let others worry about dem-
onstrating that what we do is ethical.
3.1 Investigators are Already Committed
to High Ethical Standards
One reason investigators should engage in ethical
a­ ssessment of their work is that they have already com-
mitted themselves to high ethical standards in their use
of animals. Biomedical animal research is founded on an
enormously important ethical principle (which is dis-
cussed in detail later in the chapter): It is among the most
noble and imperative of human endeavors to employ scientific
research to prevent, alleviate, and cure the pain, suffering, dis-
tress, fear, anxiety, disability, infirmity, and death associated
with human disease. The decision to use animals to fight
disease is motivated by this high ethical ideal. Having
already committed themselves to engaging in one of the
most virtuous endeavors known to humankind, inves-
tigators who use animal models are logically and un-
avoidably committed to ethical behavior in all aspects of
their work, including their use and treatment of animals.
3.2 Ethical Treatment of Animals
is Required by Law
Perhaps the most immediate and direct reason in-
vestigators should want to adhere to high ethical stan-
dards is that they must follow the laws of their respective
countries relating to animal research. Many of these laws
6 1. Ethics in Biomedical Animal Research: The Key Role of the Investigatora
impose straightforwardly ethical requirements. Failure to
comply with these requirements can result in disapprov-
al of a proposed animal research project by an institution-
al animal care and use committee (IACUC) or in some
countries a government official or licensing authority;
disciplinary action, such as public censure or a fine; or
in extreme cases termination of research (APHIS, 2016a;
Home Office, 2014; NIH, 2015a, Part II §8.5).
3.2.1 General Commitment to Ethical Behavior
Laws in many countries assert a general commitment
to ethics in the use and care of research animals. For
example, the 2010 European Directive on the Protection of
Animals Used for Scientific Purposes (EU Directive), which
represents the official policies of the European Parlia-
ment and the Council of the European Union, states
that “(a)nimals have an intrinsic value which must be
respected” (European Union, 2010, Preamble para. 12).
In the United States, the National Resources Council
Guide for the Care and Use of Laboratory Animals (the Guide)
(NRC, 2011) must be followed in all research on live ver-
tebrates that is funded by one of the institutes or centers
of the National Institutes of Health (NIH) [HREA, 1985,
§289d(c)]. In a section titled “Ethics and Animal Use,”
the Guide reminds investigators that they do not have a
legal right to conduct animal research. The use of ani-
mals is a privilege afforded by government, which it can
withdraw. As the Guide observes, this privilege is con-
ditioned on the use and care of research animals in ac-
cordance with ethical standards, some of which exceed
those specifically required by law:
The decision to use animals in research requires critical
thought, judgment, and analysis. Using animals in research is
a privilege granted by society to the research community with
the expectation that such use will provide either significant new
knowledge or lead to improvement in human and/or animal
well-being. (references omitted). It is a trust that mandates re-
sponsible and humane care and use of these animals. ... Ethical
considerations discussed here and in other sections of the Guide
should serve as a starting point; readers are encouraged to go
beyond these provisions. NRC (2011, p. 4)
3.2.2 Legal Requirements Reflecting General or
Specific Ethical Principles
Many laws, regulations, and government agency poli-
cies that pertain to animal research state explicitly, or re-
flect, very general ethical ideals or requirements. Underly-
ing the US Animal Welfare Act (AWA, 2013), for example,
is the general principle of biomedical animal research
ethics that investigators must minimize pain or distress
experienced by animal subjects. The EU Directive requires
that any harm caused to animals in a research project
must be justified by expected benefits of the research
[European Union, 2010, Art. 38 para. 2(d)]. A well-known
A. Ethics, Resources, and Approaches
set of nine general ethical principles for biomedical ani-
mal research (some of which are ­included in substance in
the fundamental ethical principles ­presented later in the
chapter) is enunciated in the US Government Principles for
the Utilization and Care of Vertebrate Animals Used in Test-
ing, Research, and Training (US Principles) (OLAW, 2015b,
pp. 4–5), which must be followed by investigators con-
ducting research governed by the Health Research Ex-
tension Act of 1985 (HREA) and its incorporated Public
Health Service Policy on Humane Care and Use of Laboratory
Animals (PHS Policy) (OLAW, 2015b, para. I). Certain laws
state more specific ethical rules, such as the requirement
of the US AWA [AWA, 2013, §2143(a)(3)(B)] and the AWA
Regulations [AWAR, 2015, §2.31(d)(1)(ii)] that investiga-
tors consider alternatives to procedures likely to produce
pain or distress in research ­animals. Some laws state even
more specific ethical requirements, such as the require-
ment of the AWA that, unless scientifically necessary, ani-
mals should not be subjected to ­multiple major invasive
procedures [AWA, 2013, §2143(a)(3)(D)].
3.3 Ethical Treatment of Animals Promotes
Sound Scientific Results
As is discussed in this chapter, among the central prin-
ciples of biomedical animal research ethics are that ani-
mals should not experience more pain or distress than
necessary for the scientific aims of an experiment, and
that they should be provided housing environments and
general care that are appropriate for their species and
contribute (consistent with experimental aims) to their
health and comfort. Studies have long established that
unnecessary animal pain or distress during experiments
can affect and invalidate experimental results (Russell
and Burch, 1959), and that removal of stressors from
housing environments can improve the validity and re-
peatability of data (Garner, 2005; Weed and Raber, 2005).
3.4 Ethical Treatment of Animals is Required by
Funding Bodies
Government agencies that fund animal biomedical
research require adherence to ethical standards as a con-
dition of support. In the United States, applicants to the
NIH must certify that projects will meet the ethical re-
quirements of the Vertebrate Animals Section of the grant
application (OLAW, 2016b), which includes the pain and
distress minimization principle (discussed below) and
justification of the use of animals and the proposed spe-
cies. Moreover, no NIH-funded animal research may be-
gin, or continue, without determination by the IACUC
that the project complies with the ethical standards of
the PHS Policy (OLAW, 2015b, para. II), and of the Guide
[OLAW, 2015b, para. IV(A)(1)]. Animal research funded
 3 Why Investigators Should Care About Biomedical Animal Research Ethics
by the National Science Foundation must also meet the
standards applied by the PHS Policy to NIH-funded ani-
mal research (OLAW, 2015a).
Private funding bodies also require grant recipients
to meet various ethical standards. For example, the
­American Cancer Society (ACS, 2016, p. 6), the American
Epilepsy Society (AES, 2016), and the American Heart
Association (AHA, 1985) require applicants to certify
that animal research projects have received approval
from their IACUC, which in the United States includes a
determination that all legally required ethical standards
will be followed. These organizations also specifically
require adherence to the PHS Policy and its ethical stan-
dards. In the United Kingdom, major funding organi-
zations, including the Association of Medical Research
Charities (AMRC, 2015), the Biotechnology and Biologi-
cal Sciences Science Council (BBSRC, 2013), the Medical
Research Council (MRC, 2016), and the Wellcome Trust
(2016) require that animal research be conducted only
when no feasible nonanimal alternative exists; that ap-
plicants employ the “3Rs” of replacement, reduction,
and refinement (discussed below), and that appropri-
ate Home Office licenses and certificates are followed.
These organizations also require grantees to follow the
ARRIVE Guidelines of the UK National Centre for the
Replacement Refinement & Reduction of Animals in Re-
search (NC3Rs, 2010), which are intended to assist inves-
tigators in preparing manuscripts submitted for publica-
tion and are considered below.
3.5 Ethical Treatment of Animals is Required
by Peer-Reviewed Journals
Virtually all journals that publish reports of animal
research require that authors state, when submitting
manuscripts or sometimes in the manuscripts them-
selves, that they have adhered to specified ethical stan-
dards. These journals always include in such standards,
observance of all applicable laws and approval of the
research by the investigator’s IACUC or other legally
required review body or government official. As noted
earlier, these laws contain fundamental principles of
biomedical animal research ethics. Some journals in-
clude ethical standards adherence to which might not
be legally required in given cases. For example, al-
though the HREA requires that NIH-funded projects
follow the ethical standards of the PHS Principles and
the Guide, many journals require adherence to the ethi-
cal standards in these documents for all research on live
vertebrates, however funded. Some journals mention
specific ethical requirements already implied by the
general requirement of following all laws and govern-
ment agency policies. Such journal requirements typi-
cally include that the minimum number of animals was
A. Ethics, Resources, and Approaches
7
used (e.g., Journal of Neuroscience; PLoS One) and that
animal pain, suffering, or distress were prevented or
minimized (e.g., Neuroscience; Neuroscience Research;
PLoS One). Some journals require adherence to the
ethical standards of the professional association under
whose auspices these journals are edited. The Journal
of Neuroscience, for example, requires that all reported
animal experimentation be conducted in accordance
with the Society for Neuroscience Policies on the Use of
Animals in Neuroscience Research (SfN, 2016), which man-
date observance of the PHS Policy and the Guide for all
experiments on live vertebrate animals.
3.5.1 The ARRIVE Guidelines
The ARRIVE Guidelines (NC3Rs, 2010), mentioned
earlier, contain a checklist of 20 directives for the prepa-
ration of manuscripts reporting animal research projects.
The Guidelines are widely regarded as an effective ve-
hicle not just for promoting standardized, transparent
descriptions of animal use that can be evaluated by the
scientific community, but also for facilitating the use of
animals in ethically as well as scientifically sound ways
(du Sert, 2011; Eisen et al., 2014; Reynolds et al., 2012; Rice
et al., 2013.) Among the directives of the Guidelines are
that the investigator “(e)xplain how and why the animal
species and model being used can address the scientific
objectives and, where appropriate, the study’s relevance
to human biology” (NC3Rs, 2010, para. 3b); “(p)rovide
details of the animals used, including species, strain, sex,
developmental stage (e.g., mean or median age plus age
range) and weight (e.g., mean or median weight plus
weight range)” (para. 8a); “(p)rovide details of housing
(type of facility, e.g., specific pathogen free (SPF); type of
cage or housing; bedding material; number of cage com-
panions; tank shape and material etc. for fish)” (para.
9a); “(e)xplain how the number of animals was arrived
at. Provide details of any sample size calculation used.”
(para. 10b); “(p)rovide details of the statistical methods
used for each analysis” (para. 13a); “(d)escribe any im-
plications of your experimental methods or findings for
the replacement, refinement or reduction (the 3Rs) of the
use of animals in research” (para. 18c); and “(c)omment
on whether, and how, the findings of this study are likely
to translate to other species or systems, including any
relevance to human biology” (para. 19).
At the time of this writing, more than 600 journals
have recommended that authors follow the ARRIVE
Guidelines (Cressey, 2016). However, relatively few
journals require compliance, and one study found that
few investigators followed the Guidelines even when
encouraged to do so (Baker et al., 2014). In light of the
support of the ARRIVE Guidelines by journal editors, it
seems inevitable that all high-impact journals eventu-
ally will make compliance mandatory. It is, therefore,
8 1. Ethics in Biomedical Animal Research: The Key Role of the Investigatora
at the very least prudent for investigators to become
used to incorporating the Guidelines into their experi-
ments and manuscript preparation. More importantly,
following the ARRIVE Guidelines will assist investi-
gators in conducting scientifically and ethically sound
experimentation.
3.6 Ethical Treatment of Animals is Essential
for Public Support
A very important reason investigators should be
sensitive to ethical aspects of their work is that without
public support most biomedical animal research would
cease. The great majority of this research is funded by
government agencies (FASEB, 2016a), which receive the
money they disburse from the public. Moreover, under-
lying government acceptance and regulation of animal
research is the general view of society that biomedical
animal research is valuable and is conducted ethically.
Were the public to reject this view, it would likely de-
mand an end to government funding, and perhaps to
biomedical animal research altogether.
Public approval of biomedical animal research can-
not be taken for granted and in some countries may be
declining. Polls conducted by the Pew Research Cen-
ter (2015, p. 141) found that in 2009, 52% of US adults
“favored,” and 43% “opposed,” “the use of animals in
scientific research.” By 2014, those in favor dropped to
47%, and those opposed rose to 50%. The annual Gal-
lup Poll on moral attitudes of Americans (Gallup, 2016)
found that in 2016, 53% of the public believed that “med-
ical testing on animals” is “morally acceptable” and 41%
that it is “morally wrong.” In 2006, the percentages were
61 and 32%, respectively.
Governments worldwide have recognized that public
support for animal research depends upon the public’s
knowledge that experimental animals are treated in ac-
cordance with high ethical standards. For example, in
enacting the 1985 amendments to the AWA (2013), the
US Congress declared (Public Law 99-198, 1985) that
“measures which help meet the public concern for labo-
ratory animal care and treatment are important in assur-
ing that research will continue to progress.” In passing
the HREA (1985) Congress stated (House Conference
Report, 1985, p. 747) that although humane standards of
animal care ”will change in the future as science advanc-
es ... the consultation inherent in the review of the care
and treatment of animals by animal care committees is
essential. Within such a framework, the public can have
confidence that the proper sensitivity, whatever the sen-
sitivity may be, to the care and treatment of animals will
occur.” The EU Directive recognizes the importance of
“the ethical concerns of the general public as regards the
use of animals in procedures” (European Union, 2010,
Preamble para. 12).
A. Ethics, Resources, and Approaches
3.7 Ethical Treatment of Animals
is the Right Thing to Do
Finally, and surely most importantly, investigators
should conduct ethical animal research because this is
simply the right thing to do. If animals are to be used
to understand human disease, they should always be
used for ethically sound reasons and in ethically ap-
propriate ways. We owe this to them, as well as to
the humans who benefit from their use in research
and who should rest assured that this use is ethically
­appropriate.
4 ASPECTS OF ANIMAL USE AND CARE
RELEVANT TO BIOMEDICAL ANIMAL
RESEARCH ETHICS
Most issues in biomedical animal research ethics per-
tain to the conduct of individual animal research proj-
ects, that is, to how experiments or research procedures
are designed and executed. Ethical questions can arise
in all stages of a project, beginning with whether there is
sufficient rationale for the use of animals or a particular
species, to whether the project treats the animals appro-
priately during experimental procedures, to when and
how the animals’ lives may or should be ended, to what
should be done with animals that survive experimenta-
tion. Although such questions typically arise in the con-
text of particular projects, biomedical animal research
ethics sometimes address them generally. Thus, one can
ask when certain kinds of physical restraint might be
unacceptable for certain species, or when it is ethically
acceptable to withhold pain relief from animals (Tan-
nenbaum, 1999). Answers or general approaches to such
questions may then sometimes be applied in particular
animal research projects.
Biomedical animal research ethics is also concerned
with the treatment of animals when they are not un-
dergoing research procedures. Many, perhaps most, re-
search animals spend the majority of their time housed
in facility animal quarters, before or after procedures
are conducted on them. To be sure, many animals are
never off-study, if, for example, effects on their physi-
ology or behavior while in the animal quarters are of
direct interest in the research, or if what they are fed
or how they are housed is part of the study. However,
even these animals can be affected by conditions of
housing and care that are not part of the research. Ethi-
cal issues relating to housing and general care of ani-
mals include whether the size, constituent materials,
and physical layout of cages or other enclosures are
sufficient to meet certain behavioral needs, whether
animals receive adequate care from veterinarians and
support staff to address health or welfare problems,
Another random document with
no related content on Scribd:
quexas en vuestra presencia; no
que yo, señora, de vos me quexe
ni Dios lo quiera, que no deuo
más para que las pasiones que
con mis deseos me aquexan
sepays, por merito de las quales
os suplico que no medido lo que
yo en respecto vuestro me
merezco, mas considerado lo que
por haueros visto e desear ser
vuestro padezco, por tal señora
me acepteys; no para dar más
bien a mi mal de consentir que yo
señora por vuestro seruicio lo
padezca, por que ni más osaria,
señora, pedir, ni tanto me
atreueria creer merecer.

BELISENA
Muchos dias ha, Flamiano, que
conozco en tus meneos lo que el
desuario de tu pensamiento te ha
puesto en la voluntad; e no creas
que muchas vezes dello no haya
recebido enojo, e algunas han
sido que me han puesto en
voluntad de dartelo a entender,
sino que mi reputacion e
honestidad me han apartado
dello, e aun en parte el respecto
de la buena figura en que tu
discrecion hasta agora he tenido.
Mas pues que tu atreuimiento en
tal estremo te ha traydo, que en
mi presencia tu fantasia hayas
osado publicar, forçado me será
responderte, no lo que dezirte
queria segun mi alteracion, mas
segun la vanidad de tu juyzio
merece. Lo qual aunque consejo
te parezca deues tomar por
reprehension; e digo que no te
acontezca semejante
pensamiento poner en parte
differente de ti, donde no puedas
menos hazer de verte cada hora
en infinitas necessidades e al fin
sin ver cabo á lo que desseas,
que lo hayas de ver de tu vida y
de tu honrra. Mas razon seria que
primero ygualasses la medida
donde bastas llegar con el
merecer, que no que publicasses
do querrias subir con el dessear e
aun alli, segun se suele, hallarás
tarde el contentamiento que el
deseo querria.

FLAMIANO
Mis ojos, señora, que de mis
males han sido la causa, no
tuvieron juyzio más de para
miraros e ver las perficiones que
Dios en vos puso, para que
viendoos pusiesen mi corazon en
el fuego que arde; llegada alli
vuestra figura, no pudo menos
hazer de lo que ha hecho. Mi
saber no pudo ser tanto para
temer los inconuenientes de mi
daño que vuestra hermosura no
fuesse más para causallo sin
poder ser resistido. Pues llegado
aqui mi pensamiento determinose
en que lo mucho que el merecer
desyguala mi pena del desseo,
las sobras della misma son tantas
que lo yguala todo, pues que,
señora, mi intencion no os pide
mas de licencia para padescer,
que desta suerte cierto no puede
ser reprouada pues que no es
mala. Ansi que, señora, pues que
tanto la virtud y nobleza en vos
sobra, no useys comigo por el
rasero de la crueza, pues que
mudarse ya mi cuydado es
imposible. E assi de vos no quiero
consejo; remedio es el que pido
pues que no le puedo esperar
sino de vuestra mano.

BELISENA
No creas tú, Flamiano, que la
pasion o males que publicas que
sientes, a mí dellos me plega,
ante en muchas maneras dello
me pesa. Lo vno es que á mi
causa siendo en mi perjuyzio tú
los padezcas. Lo segundo que te
atreues á ponerte en ello y aun
publicarlo. De suerte que en
muchas maneras me enojas y en
más me harias plazer y servicio
que dello te dexases. Y esto seria
seruirme como dizes que
desseas; para esto que te digo,
como ya te he dicho, los
inconuenientes de mi estado y de
mi condicion y honestidad me dan
inconueniente no solo para que
como hago dello reciba mucho
enojo, mas para que tú aunque
mill vidas como dizes perdiesses
yo dellas haya de hazer ni cuenta
ni memoria. Assi que lo mejor
será que desto te apartes e en
esto me harás seruicio como
dizes que desseas y aun me
ternas haziendolo contenta; e
pues que tanto mio eres, segun
dizes, yo te mando que lo hagas,
porque quites tu vida de peligro e
aun a mí de ser enojada.

FLAMIANO
Quando, señora, la pena
verdadera de amor como es la
mia está sellada en el alma, pues
que justa razon alli la haya
puesto, en el coraçon está
imprimida de suerte que sin él e
sin ella no pueda salir de alli.
Pues ¿como quereys, señora,
que mi cuydado se mude?, que el
dia primero que os vi, dentro en
mis entrañas e coraçon quedó el
propio traslado vuestro
perfectamente esculpido, e
despues aca quantas estradas
me haueys tirado que son
infinitas, llegadas alli, el fuego que
en tal lugar hallan las funde,
porque son de oro siendo
vuestras e fundidas hallan alli
vuestra effigia e de cada vna
dellas se haze vn otra semejante.
Assi que aunque el coraçon y el
alma con las principales
sacassen, el cuerpo quedaria
lleno con tantas que de aqui a mill
años en mi sepultura se hallarian
dellas sin cuento, e aun en todos
mis huessos se hallaria vuestro
nombre escripto en cada vno.
Ansi, que señora, si quereys que
de quereros me aparte, mandad
sacar mis huessos e raer de alli
vuestro nombre, e de mis
entrañas quitar vuestra figura,
porque ya en mi está conuertido
en que si alguno me pide quien so
digo que vuestro. E si esto a
desuario se me juzgasse, mayor
lo haria quien tal quissiese juzgar,
porque no hay nayde que con mis
ojos, señora, os mire que no
conozca ser justo lo que hago; e
como ya he dicho, aunque en la
razon mia encobrir lo quisiesse no
puedo, porque el fuego de dentro
haze denunciar a la lengua la
causa. Pero pues que en vuestra
mano está matarme o darme la
vida, e pues que della teneys la
llaue, ved vos si lo podeys hazer
e ganareys la victoria del tal
vencimiento. E si con quitarme la
vida pensays acabarlo, dudolo,
porque aunque del coraçon e las
otras partes vos apartassedes
con matarme, ni mas ni menos en
el alma os quedariades, de do
jamas os podreys quitar porque
es inmortal a causa de estar vos
en ella. E si de mi se partiesse
donde agora mis passiones la
tienen presa y atormentada,
jamas de vuestra presencia se
partiria, donde con mucho
contentamiento estaria contino.
Assi que si agora estando comigo
os enoja ausente, mira que hará
entonces estando presente, e
bien sé que pues agora os
enojays por seros yo de mi grado
captiuo, que despues de yo
muerto más enojo recibireys de
vos matadora, e sola esta gloria
que de mi muerte se espera me
basta a mi para que contento
pierda la vida, pues que con ello
yo seré fuera de pena e vos con
pesar arrepentida. Podreys
señora dezir entonces que no es
vuestro el cargo sino mia la culpa
pues que yo mesmo me lo he
buscado y querido mi daño contra
vuestra voluntad. Entonces mi
alma os negará la partida
diziendo: no, no, no es ansi, que
el cargo, señora, tuyo es pues
que tan cruelmente tan mal le
trataste no pidiendote más bien
de licencia para sofrir su mal sin
ninguna offensa tuya ni más gloria
suya.
 BELISENA
Si sofrirte lo que faces me
offende, oyrte lo que dizes me
perjudica y enoja; ¿qué hará
responder a la vanidad de tus
razones? Yo te he ya dicho lo que
te cumple, bastarte deue para no
esperar mas disputa en este caso
de lo que te conuiene. No delibero
mas sobre ello hablarte, porque
creo que tu discrecion te hará
determinar lo que te cumple. Los
mios vienen, quedate con Dios y
creeme haziendo lo que te tengo
dicho.

FLAMIANO
Digo, señora, finalmente que no
puedo porque ni mi voluntad a
ello no puede doblarse, ni mi
querer puede dello quitarse, e
aunque aquí tan solo de bien e
tan acompañado de pesar me
dexeis, digo que allá donde vos
vays, allá voy, y aunque vos vays,
aqui quedays donde yo quedo,
porque ni allá, ni acá, ni en
ninguna parte donde yo me halle,
nunca vuestra vista de mis ojos
se quita, sino que en mi fantasia
do quiera que esteys, do quier
que esten, los dos juntos
estamos. E si esto, señora, no
creeys, mis obras os haran dello
testigo.
Al fin la señora Belisena se partio
con Isiana e muy enojada, a lo
que mostraua, e llegó a la
compañia de los suyos. Flamiano
quedó a solas, fuesse por otra via
con el consuelo que pensar
podeys; en aquella noche todos
los caualleros cenaron con el
señor cardenal, donde se
concerto de yr venidos de la caça
a vnos baños que ocho millas de
la ciudad estan de la mar, en vn
muy hermoso lugar que Virgiliano
se llama, porque supieron que la
señora duquesa e la princesa de
Salusano con otras muchas
damas se yuan por estar alli todo
el mes de Abril, como cada año
las damas y señoras de
Noplesano acostumbran hazer.
Visto Flamiano que esta jornada
se le aparejaua conforme a su
desseo, suplicó al señor cardenal
que ordenase vn juego de cañas
para el segundo dia de pasqua
que todas las damas ya a
Virgiliano serian venidas. De lo
qual el señor cardenal, fue tan
contento que se ofrecio tener el
vn puesto con la meytad de
aquellos caualleros, desta
manera: que los de su puesto
saldrian a la estradiota vestidos
como turcos con mascaras y
rodelas turquescas, vestidos
todos de las colores que su
señoria les daria, y que jugarian
con alcanzias. E que Flamiano
tuviesse el otro puesto a la gineta
con los otros caualleros que alli
primero se hallaron en la caça. E
que ante que al puesto saliessen,
que saliessen ellos todos juntos e
començassen su juego de cañas
partidos por medio. En el qual
juego él con sus turcos llegaria
como hombre que viene de fuera,
e assi juntados ellos todos,
començarian el otro juego contra
los que en él viniessen. E ansi el
señor cardenal tomó a cargo de
suplicar a la señora princesa que
para aquella noche conbidase a la
señora duquesa e á Belisena, con
todas las otras damas que alli se
hallassen, para que en su posada
aquella noche passado el juego
todas cenassen y alli hiziessen la
fiesta. Pues acabada la caça,
dende a dos dias con mucho
plazer los vnos e los otros todos
juntos a la ciudad se tornaron.
Donde despues de llegados,
Flamiano acordo de enbiar a
Felisel a visitar a Vasquiran con el
qual acordo respondelle a su
carta. E despachado que le houo,
Felisel se partio, e llegado a
Felernissa donde halló a
Vasquiran, despues de hauer
hablado mucho con él en especial
de las cosas dela caça e lo que
en ella se era seguido, la carta de
Flamiano le dió, la qual en esta
manera razonaua.

CARTA DE FLAMIANO Á
VASQUIRAN EN RESPUESTA
DE LA SUYA POSTRERA
No quiero, Vasquiran, dexarme de
responder a tus cartas e quexas,
si quiera porque no pienses que
razon me falta para ello, como a ti
crees que te sobra para lo que
hazes. Yo, si bien me entiendes,
no digo que de la muerte de
Violina no te duelas como es
razon que lo hagas, mas que los
estremos dexes e apartes de ti,
pues que in genere son
reprobados; porque como ya te
he dicho y tú dizes, tus lastimas
todas la muerte las ha causado, y
en verdad al parecer estas son
las mas crudas de sofrir, y al ser
las mas leues de conortar, pues
como dicho tengo, el tiempo e la
razon naturalmente las madura e
aplaca de tal suerte que assi
como la carne muerta en la
sepultura se consume, assi el
dolor que dexa en la viua se
resfria. Porque si assi no fuesse,
muchas madres que
ardientemente los hijos aman e
los pierden, por ser fragiles para
soffrir el dolor con la braueza dél,
con la flaqueza de la complision,
si este remedio el tiempo
naturalmente no les pusiesse, las
mas dellas del seso o de la vida
vernian a menos, e aun algunos
padres lo mismo harian, e otras
muchas personas que de
conjunto amor contentos
acompañados viuian como tú
hazias. Empero como he dicho el
natural remedio lo remedia
continuamente, e donde este
faltasse o si assi no fuese, digo
que por razon más obligado
serias segun quien eres a hazer
lo que digo que lo que hazes, por
muchas causas que ya te tengo
dichas, porque como sabes, la
estremidad del plañir nace de la
voluntad, la virtud del soffrir es
parte de la razon.
Pues mira quan grande es,
nuestra differencia entre la
voluntad é la razon. Lo vno parte
de discrecion e cordura; lo otro o
es o está a dos dedos de locura,
en especial que los virtuosos
varones más son conocidos en
las aduersidades por su buen
seso e sofrimiento que no en las
prosperidades por grandezas ni
gouierno; porque lo vno muchos
respectos lo pudieron causar para
hazerse, lo otro sola virtud lo
templa para sofrirse. Assi que por
todas las partes verás que por
fuerça tu dolor ha de menguar.
Mas ¿qué hare yo que si sola vna
vez que vi a la que mi mal ordena,
de tantos malos me fue causa?
en las otras que la veo ¿qué
puedo sentir? Su ausencia me
atormenta de passion; su
presencia me condena de temor;
su condicion e valer me quitan
esperança; mi suerte y ventura
me hazen desconfiar. Mi pena me
da congoxa incomportable. Lo
que siento me haze dessear la
muerte; remedio en mi no le hay;
della no se espera. E assi tengo
más aparejado el camino de
desesperar que abierta la puerta
de esperança para ningun bien.
Assi que por Dios te ruego que
comiences á poner consuelo en ti,
porque puedas presto con tu
compañia venir a poner remedio
en mí, y con tal confiança me
quedo cantando este villancico
que a mi proposito haze y a mi
pesar he hecho.

Yo consiento por seruiros

mi muerte sin que se sienta
vos señora no contenta.
El primer dia que os vi
tan mortal fue mi herida
que en veros me vi sin vida
y el viuir se vio sin mi,
pues que en viendoos
consenti
mis males que son sin cuenta,
vos señora mal contenta.
 Consenti verme sin ella
solamente por miraros
y por solo dessearos
tuue por bueno perdella;
y más que los males della
quise qu'el alma los sienta
y vos dello descontenta.
Consenti que mi tormento
tan secreto fuese y tal,
que el menor mal de mi mal
diesse muerte al sentimiento;
quise más qu'el soffrimiento
que lo suffra y lo consienta
por hazeros más contenta.
De suerte que mis sospiros
aunque sean sin compas
los quiero sin querer mas
de quereros y seruiros,
sin más remedio pediros
de la muerte que m'afrenta
que veros della contenta.

LAS COSAS QUE VASQUIRAN

CONTO A FELISEL
DESPUES DE LEYDA LA
CARTA, QUE LE HAUIAN
SEGUIDO YENDO A CAÇA
Despues de leyda Vasquiran la
carta que Felisel le dió, hablando
de muchas cosas Felisel le conto
todas las cosas de la caça, assi
de los caualleros y damas que en
ella fueron como de los atauios
que todos sacaron, e aun parte de
lo que su señor con Belisena
passó hablandose con ella a
solas. Pues hauiendolo todo muy
bien relatado, otro dia
paseandosse los dos como otras
vezes solian por vna sala,
Vasquiran le començo á dezir:
Pues que ayer, Felisel, me
contaste todos los mysterios de la
caça que allá haueys tenido, e
aun lo que a tu señor en ella le
siguio, quiero contarte lo que a mi
en otra me ha acontecido.
Flamiano, como dizes, fue por
acompañar a quien de
enamorados pensamientos
acompañado le tiene e aun por
dar con su vista descanso a sus
ojos. Yo por acompañar a mi
soledad de mas soledad e por dar
a los mios con ella de lagrimas
más compañia con menos atauios
e mas angustias la semana
passada tambien me fuy á caça,
en la qual me acontecio lo que
agora oyras.

RECUENTA VASQUIRAN Á
FELISEL LO QUE LE
ACONTECIO EN LA CAÇA, E
LA OBRA QUE SOBRE ELLO
HIZO
Estando con sus canes estos mis
seruidores en sus paradas
puestos como yo los hauia
dexado, contecio que vn ciervo e
vna cierva juntos en la vna dellas
dieron, de que dadas laxas a los
perros començaron a seguirlos
por vna llanura que entrellos e un
bosque se hazia. E siendo los
canes muy buenos dieronles vn
alcance en el cual la cierua se
houo de apartar de su compañia e
vino a dar donde yo estaua, por
su desventura e la mia, e assi
como yo la vi venir salile por el
traues adelante e ante que al
bosque llegasse la maté.
Llegados alli parte destos mis
seruidores, porque ya era algo
tarde mandela cargar sobre vna
azemila con la otra caça que
muerto hauiamos, y yo comence
a venirme la via de aquella
eredad mia a donde la otra vez
me hallaste, e seyendo ya al
aquanto del bosque alongados,
sentimos los mayores bramidos
del mundo, los quales por nos
oydos, paramos por saber qué
podria ser, e vimos venir vn cieruo
que en el bosque se nos era
entrado bramando, y era el que
en compañia de la cierua venia, el
qual ni por el temor de los canes
que al encuentro le salieron, ni
por lo que los mios le ocuparon
jamas dexó de hazer su via hasta
llegar al azemila do la cierua
venia cargada. E como yo lo vi
pense lo que podia ser como fue,
aunque milagro parezca, e assi
mandé que ninguno le hiziesse
daño. Pues llegado que fue do su
dolor lo guiaua, començo á dar de
nuevo muy mayores bramidos
derramando de los ojos infinitas
lagrimas. Como tal le vi hazer
tanto dolor, començo a refrescar
en mi llaga, que temiendo en mi
algun desmayo que afrenta me
hiziesse, mandé lo dexassen
estar e segui mi camino para
donde él yva, mas como nos vido
partir, con mayores gemidos
començo a seguirnos hasta llegar
do yo yva, de donde jamas se es
partido. Como esto vi mandé que
a la cierua desollassen el cuero e
lo hinchiessen de feno e dentro
en el jardin lo colgassen en vna
lonja que en el hay tan alto que el
ciervo solamente pudiesse
alcançar a su cabeça. E desde
aquel dia que alli lo pusieron
mandé meter dentro al cieruo e
jamas de donde la cierua está se
es partido, saluo cuando
costreñido de la hambre algun
poco por la huerta a pacer se
aparta. Pusome tanta tristeza ser,
Felisel, lo que te he contado, que
despues de hauer cenado a solas
retraydo en mi camara,
veniendome a la memoria todas
mis glorias pasadas y la congoxa
presente, juzgando por lo que
este irracional hazia lo que de
razon yo deuia hazer, con infinitas
lagrimas comence contra mí
maldiziendo mi desuentura a dezir
infinitas e muy lastimeras
palabras, tantas que largo seria
contarlas. Saluo que estando assi
yo me senti assi venir a menos el
sentido e no sé si trasportado del
juyzio o si de dolor y del sueño
vencido, yo vi en vision todas las
cosas que a tu amo embio dentro
en una carta que le tengo ya
escripta, lo qual verás en versos
rimados conpuestos más como
supe que como deuiera o
quisiera. E despues hize sobre
este caso deste cieruo esta
cancion, la qual no he querido
que tu amo la vea, por que no
halle en ella con que responder a
mi carta como suele.

¿Que dolor puedo quexar

de mis angustias e males
viendo que los animales
mayor sienten mi pesar?
Quexaré de mi dolor
que es tan crudo su tormento
que vn bruto sin sentimiento
le siente mucho mayor,
de pesar que yo le siento,
mas no se puede ygualar
con mis angustias mortales
porque ell alma de mis males
mayor siente mi pesar.

Acabado que houo de decirle la

cancion le dixo: Felisel, yo querria
que mañana te partiesses, porque
llevasses a Flamiano vn cauallo
mio de la gineta con vn gentil
jaez, que agora poco ha me han
traydo de España, porque
aproueche para el, pues que a mí
ya seruir no me puede. Querria
que llegasses a tiempo que para
el juego de las cañas que me has
dicho le siruiesse. Otro dia
recebido Felisel el cauallo e la
carta se partio. E llegado a
Noplesano, halló que Flamiano
con todos los caualleros eran ya
partidos para Virgiliano, porque la
señora duquesa e la princesa con
todas las damas ya estauan alli.
Donde otro dia Felisel llegó, con
el qual Flamiano holgó mucho e
houo mucho plazer de oyrle
contar lo que a Vasquiran hauia
acontecido e tambien con el
cauallo que era muy bueno y el
jaez muy rico, en especial
llegando a tal tiempo. Y recebida
la carta començola a leer la qual
assi dezia.

CARTA DE VASQUIRAN Á
FLAMIANO EN RESPUESTA DE
LA SUYA
Quanto mejor seria, Flamiano,
que a esta question pusiessemos
silencio que no proseguirla, pues
que tan poco prouecho a los dos
nos acarrea. Tú me dizes que no
me reprueuas porque de mi mal
me duelo pues que es razon que
lo haga, sino que no deuo tanto
en estremo dolerme. Mi mal
quisiera yo que limitaras que no
fuera tan grande, que mi tristeza
pequeña es para con él. Dizes
que como la carne muerta en la
sepultura se consume, assi el
dolor que dexa en la viua se
resfria; falso es esse argumento
pues en mi que lo prueuo por el
contrario lo veo. Tornasme a
alegar las mugeres que perderian
el sentido si por esto no fuesse. A
la fe por ser ellas flacas de
sentido e fragiles pierden dello la
memoria, que no por lo que dizes.
Si honesto me fuesse alegarte
cosas de nuestra fe, vna cosa te
diria de la que no tuvo par, que en
tal caso hizo, con que callasses.
Tambien me alegas como
philosopho lo que de la voluntad o
de la razon parte, quál es auto
mas virtuoso, e das lexos del
terrero, que los que desso han
glossado, en especial Juan de
Mena e muchos no ponen
contraste en tal caso, entre la
voluntad e la razon, saluo de
aquellos apetitos que
viciosamente muestra naturaleza,
desseo voluntario, que el dolerse
nadie de la cosa amada de puro
amor e gratitud y contentamiento
que le tenia, le parte viendola
perdida. Pues estos autos
virtuosos y razonables son, que
no voluntad voluntaria. Ansi que
no te cale philosophia comigo que
poco te aprouecharia ni a
Aristoteles si mi mal sintiera. Mas
sabía el Petrarca que no tú ni yo,
mas ya sabes lo que respondio
siendo juzgado porque a cabo de
veynte años que madama Laurea
era muerta la plañia e la seruia,
quando dixo: ¿Que salud dió a mi
herida quebrarse la cuerda del
arco? Nunca de tu mal vi ningun
martir e del mio verás todas las
poesias y escripturas dende que
el mundo se començo hasta
agora llenas, de lo que aun la
sangre del martir Garcisanchez
viua tenemos e no oluidada la del
mesmo Petrarca que te he dicho,
sin otros infinitos que dellos no se
escriue. Tú no hallas remedio
para ti que cada dia hablas o
puedes hablar a quien te pena;
quieresle hallar para mi que no le
tengo. Tambien me dizes que la
primera vista tanto tanto mal te
causó, ¿que sentiras en las
otras? Digo que la primera vez te
enamoró, las otras te
reenamoran, todo el mal que te
causa su ausencia es desseo de
verla. El que te haze su presencia
es desseo de codiciarla. En fin,
son vanidades que la vna con la
otra se texen; mas si lo quieres