Forensic Science International: Animals and Environments 1 (2021) 100025

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Technical Note

Morphological analysis: A powerful tool in wildlife forensic biology

Pepper W. Trail
National Fish and Wildlife Forensic Laboratory, 1490 E. Main Street, Ashland, OR, 97520, USA

A R T I C L E I N F O A B S T R A C T

Keywords: It is a truism that “structure is the first thing that we notice whenever looking at organisms” (Gans 1985), and the
Wildlife forensics description and analysis of structure is the province of the science of morphology. Morphological analysis is a
Morphology well-established and cost-effective technique for the taxonomic identification of wildlife remains. Despite this, it
Identification
is under-utilized in wildlife crime investigations for reasons including a shortage of trained specialists, the
Comparative anatomy
Taxonomic assignment
challenge of accessing reference specimens, and the perceived greater rigor of DNA analysis. This paper reviews
Class characters the methodology of morphological analysis, addresses perceived challenges, and demonstrates the effectiveness
of the technique in wildlife forensic biology casework at the National Fish and Wildlife Forensic Laboratory of the
U.S. Fish and Wildlife Service.

Introduction
An essential step in wildlife crime investigations is the identification
of the species represented in the evidence. Some species have no legal
status, while others are strictly protected, with potentially severe pen­
alties for killing or commercial trade. Therefore, taxonomic identity
must be established to determine if a wildlife crime has been committed.
Identification of the species present in wildlife evidence is typically
made either by morphological or DNA analysis. In morphological
analysis, trained and competency-tested specialists make species iden­
tifications based on physical characters present in the evidence, such as
avian plumage patterns, mammal dentition, or reptile scale counts.
Evidence items are compared to verified reference specimens and/or
authoritative data sources, and taxonomically informative characters
are documented (for an excellent review of this process in the
zooarchaeological context, see [2]).
This process of morphological comparison is formalized in field
guides, technical keys, and taxonomic monographs, and is routinely
used for species identification in a wide variety of disciplines, including
paleozoology, botany, entomology, ichthyology, herpetology, ornithol­
ogy, and mammalogy. Indeed, morphological characters formed the
exclusive basis for species definitions from the time of Linnaeus until
recent decades. The detailed morphological “diagnosis” remains a
required and essential element for new species descriptions, even as
behavioral, ecological, and genetic characters are now often included
(for a recent example, see [3]). Identifications of wildlife evidence by
morphological examination are routinely admitted as evidence in US
courts, demonstrating the acceptance of the method by the legal system.
Despite the wide use of morphological analysis for species identifi­
cation in a variety of contexts, it is under-utilized in wildlife forensic
biology relative to DNA analysis. Few wildlife forensic facilities employ
morphologists or maintain collections of reference specimens. As will be
discussed below, one reason for this is a shortage of qualified morpho­
logical analysts. However, there also appears to be a common precon­
ception among non-scientists, including attorneys, that DNA analysis is
more rigorous and accurate than identifications based on morphology.
This paper will document the utility of morphological analysis as a
rigorous and cost-effective tool in wildlife forensics, especially in part­
nership with DNA.
Morphology: analysis in evolutionary context
A clear distinction must be made between morphological analysis
and mere description. In the broad field of forensics, confusion may arise
if morphology is considered to be equivalent to the examination of non-
biological materials, such as bullets, paint chips, tool marks, and tire
impressions. Examination of non-biological materials is limited to
description and comparison to a series of artificial (human-produced)
exemplars. This does not allow hypothesis testing beyond successive
comparisons in search of a match.
While the comparative method is basic to both morphology and to
the forensic examination of non-biological materials, morphological
comparison of wildlife samples occurs within an evolutionary frame­
work. The documentation of morphological characters in relation to
evolutionary phylogeny and with regard to adaptive function is central
to taxonomy and an array of other disciplines, including anatomy,

E-mail address: pepper_trail@fws.gov.

https://doi.org/10.1016/j.fsiae.2021.100025
Received 3 December 2020; Received in revised form 8 July 2021; Accepted 26 July 2021
Available online 31 July 2021
2666-9374/Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
P.W. Trail
physiology, and paleontology [1,4–12].
Morphological analysis in biology proceeds through the hierarchy of
taxonomy, from order to family to genus to species, with shared derived
characters (synapomorphies) noted at each taxonomic level. For
example, a whole trophy mount of a polar bear (Ursus maritimus) can be
identified to order Carnivora based on its carnassial teeth and robust
claws, to the family Ursidae by its size, body shape, and plantigrade feet;
and to species by its all-white pelage and well-furred pads of the feet.
In practice, complete animal remains comprise a small fraction of
wildlife evidence, and characters distinctive for particular taxonomic
levels are often missing. A bear rug, an eagle feather headdress, and a
crocodile skin handbag lack most characters of the whole animal.
Moreover, counterfeit, modified or synthetic objects are regularly sub­
mitted for identification in wildlife crime investigations. For example,
“tortoiseshell” objects are often fabricated from plastics or resins, not
actual sea turtle scutes. Carved bone is a frequent substitute for elephant
ivory. Legally-traded feathers of turkey (Meleagris gallopavo) may be
dyed and trimmed to simulate the feathers of protected eagle species.
Despite these challenges, the taxonomic expertise and evolutionary
perspective of the morphologist allows the available physical characters
to evaluated in the context of phylogenetic relationships. The experience
of the morphologist – along with access to a comparative specimen
collection and extensive familiarity with the relevant scientific literature
– is critical for assessing and documenting informative characters, and
for determining when genetic analysis is required for identification.
The scope of morphological analysis in wildlife forensics:
identification, not individualization
Another source of confusion about the role of morphology in foren­
sics arises from inconsistent usage of the terms “identification” and
“individualization” in human vs. wildlife forensics [13].
“Identification” as performed by wildlife forensic scientists is anal­
ysis to determine the species (or higher taxonomic classification) of the
questioned evidence. This is the typical request from investigators in a
wildlife crime case, who need to know if a protected species was taken,
but not which individual animal was involved. Shared morphological
characteristics are used by scientists to define taxonomic groups, and
these class characters can be reliably associated with evolutionary line­
ages down to the species level, based on exhaustive research by spe­
cialists in the relevant groups. For example, the plumage pattern of a
Scarlet Macaw (Ara macao) is a class character diagnostic for that spe­
cies, thus allowing conclusive species identification. No other bird spe­
cies has bare white facial skin, bright red body plumage, long graduated
tail, and blue wing feathers with yellow coverts. However, this species-
diagnostic pattern does not allow one individual Scarlet Macaw to be
distinguished from all others.
“Identification” has a quite different usage in human forensics (e.g.,
[14]). In human cases, the species source of questioned biological ma­
terial (e.g., blood or hair) typically does not need to be identified – it is
known to be human. Thus, requests for “identification” of human evi­
dence seek to know the person from which the material originated – the
individual source. This is more precisely termed “individualization.”
Morphological examination of human skeletal and dental remains is a
standard technique for individualization in forensics. However, attempts
at human individualization of trace evidence based on morphological
examination have been found to be generally unreliable in comparison
to DNA analysis [15]. Fingerprint examination is the only morphological
trace-comparison technique still widely used for individualization in
human forensics, although debate about its reliability continues
[15–17]. Other morphological individualization techniques involving
trace evidence such as hair and bite marks have largely fallen from
favor.
In general reviews of forensic methodology, attorneys and judges
accustomed to working with human forensic cases thus may question
morphological identifications in wildlife forensic biology, mistakenly
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Forensic Science International: Animals and Environments 1 (2021) 100025
assuming they claim to provide individualization. This issue has arisen,
for example, in the multi-disciplinary OSAC (Organization of Scientific
Area Committees) forensic science effort currently under way in the
United States.
In fact, wildlife forensic morphologists almost never conduct indi­
vidualization analyses. While field biologists can sometimes recognize
individual animals in species with complex markings (e.g., cheetahs,
[18]; tigers, [19]; giraffes [20],), such recognition systems are tied to
particular study populations. To be useful for individualization in the
forensic context, the full range of pattern variation for the species would
need to be known, and this is rarely the case. Morphological comparison
may allow a possible individual “match” to be rejected, based on clear
pattern differences between an evidence item and a possible source, but
confirmation of an individual match will likely require DNA analysis, just
as with human individualization. Even with genetic analysis, individu­
alization is possible only for animal species with extensive population
genetics databases. In such cases, individual identity is confirmed with
appropriate statistical analysis, often reinforced with the use of likeli­
hood ratios in a hypothesis-testing framework.
Morphology in wildlife forensics – methodology, training, and
certification
Methodology of morphological examination in wildlife forensics
The Society for Wildlife Forensic Science (SWFS), the professional
organization of wildlife forensic practitioners, has adopted a series of
requirements (standards) and additional best practices (guidelines) for
morphological analysis [21], as well as for DNA analysis and chemical
analysis for timber identification. The SWFS Standards and Guidelines
for Morphology provide the best summary of methods for morphological
analysis and documentation in wildlife forensics.
The most fundamental standards are:
• The analyst shall consider the completeness and condition of the
evidence, and the presence/absence of taxonomically informative
characters.
• The analyst shall examine, interpret, and document morphological
similarities between the evidence item and specimens of known
species source and/or appropriate scientific reference material.
• The analyst shall consider the diagnostic value and inter- and
intraspecific variability of the characters being analyzed.
• Age and sex characters of the evidence shall be evaluated, and the
analyst shall determine whether available reference materials are
appropriate for correct data interpretation and species identification.
• When the evidence item does not represent a complete organism, the
analyst shall evaluate the appropriate taxonomic level to which
identification can be made.
Importance of reference collections
As the above makes clear, morphological analysis in wildlife foren­
sics relies upon access to a collection of verified reference specimens.
Published literature and authoritative online databases are important
and sometimes necessary for morphological analysis, but comparison of
evidence with physical specimens should be performed whenever
possible [22].
Reference specimens do not need to be physically located at the
forensic laboratory. Facilities conducting wildlife forensics are often in
large cities with major universities and museums. Analysts with access
to such collections can use those specimens for required comparisons,
either by obtaining specimens on loan, or comparing data and photo­
graphs of the evidence to off-site specimens. It is generally not possible
to take the evidence items themselves out the laboratory for off-site
comparison, owing to chain-of-custody requirements.
The most convenient arrangement is for the reference collection to be
P.W. Trail
located at the wildlife forensic laboratory or its host institution. The
Australian Centre for Wildlife Genomics carries out wildlife forensic
casework at its laboratory in the Australian Museum, Canberra, allowing
access to the museum’s extensive collections. The U.S. National Fish and
Wildlife Forensic Laboratory (NFWFL) is located far from major natural
history museums and maintains its own collection in Ashland, Oregon.
The NFWFL collection represents over 2450 species (approximately
1650 birds, 520 mammals, and 280 reptiles and amphibians), tailored to
the taxa most important in wildlife forensic casework. These specimens
are derived entirely from donations and from salvaged carcasses; no live
animals were killed in order to create this collection.
The development of a specimen reference collection may seem
daunting, but experience at NFWFL demonstrates that salvaged speci­
mens are available from a wide variety of sources. Confiscated items
may also provide reference specimens, once they have been verified
with a primary source. Further, collections need be only as compre­
hensive as the taxa to be identified morphologically. For example, if a
state crime lab uses morphology only to identify species of the deer
family (Cervidae) in poaching cases, then only specimens of those spe­
cies need be available – provided that age, sex, and intraspecific varia­
tion are adequately represented. These reference specimens can also
provide important voucher samples for DNA analysis, once their identity
has been verified by morphologists.
Training, testing, and certification of morphology analysts
Taxonomic identification based on morphological analysis requires
significant training and experience, to include:
• Thorough knowledge of the taxon-defining characters of the group
being examined. In wildlife forensics, most casework involves ver­
tebrates, and experts are typically mammalogists, ornithologists, or
herpetologists. Morphologists in other disciplines, such as botany
and entomology, can also provide valuable expertise in wildlife fo­
rensics when needed.
• Thorough knowledge of the diagnostic characters of the species taxa
under consideration, as well as of intraspecific variation related to
age, sex, and geographical origin.
• Thorough knowledge of the current taxonomy of the taxa under
consideration. For morphologists working in wildlife forensics, this
needs to include familiarity with the taxonomy used in the laws
governing wildlife, which often pre-dates current classifications.
A challenge for the expanded use of morphological examination in
wildlife forensics is the shortage of qualified practitioners. Much has
been written in recent years concerning the global shortage of taxono­
mists, which has serious implications for documenting, understanding,
and conserving biodiversity [23–26]. Forensic morphologists must be
taxonomic experts for the groups they examine, and thus recruiting
qualified applicants may be difficult. If wildlife forensics provides
additional employment opportunities for taxonomists, this may increase
the number of students entering the fields of taxonomy and morphology.
To be accepted as an expert witness in U.S. courts, a forensic prac­
titioner must demonstrate both advanced training and significant
experience. While a Bachelor’s degree in a relevant scientific discipline
is generally the entry-level requirement in forensics, a Master’s degree
for work on a specific taxonomic group, with at least one year of case­
work experience, is desirable for morphological analysts. In countries
where access to higher education is limited, these requirements may be
more flexible, as highly experienced individuals, such as park rangers,
may have demonstrable expertise at identifying certain species.
In both developed and developing countries, a rigorous program of
proficiency testing is essential for demonstrating analysts’ expertise.
Proficiency testing is a standard requirement for analysts in forensic
laboratories (ISO/IEC 17025 and 17043), and testing programs are
administered by a variety of organizations. SWFS operates a proficiency
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Forensic Science International: Animals and Environments 1 (2021) 100025
testing program for DNA analysts in wildlife forensic biology (http
s://www.wildlifeforensicscience.org/proficiency-testing/) involving
approximately 20 participants annually, but no comparable external
program for morphology analysts has been established. This is because
only a single U.S. wildlife forensic biology laboratory, the National Fish
and Wildlife Forensic Laboratory, has a dedicated morphology section.
With the approval of the forensic certification organization ANSI-
ASQ (ANSI-ASQ National Accreditation Board ANAB FM 3041),
NFWFL has developed an internal proficiency testing program in
Morphology administered by the Quality Assurance Manager, which
each analyst completes annually. Verified morphological specimens
with all identifying data removed are submitted for identification, and
the examiner carries out full analysis and documentation according to
casework protocols. Proficiency tests are designed to test all aspects of
an analyst’s morphological expertise over a period of years. For
example, a proficiency test in forensic ornithology may concern osteo­
logical evidence in one year, and microscopic feather examination the
next.
SWFS also administers a certification program to verify that wildlife
forensic practitioners are well-qualified for their scope of work. This
program, which requires successful participation in a proficiency testing
program and a minimum of one year of casework experience, provides
certification to analysts in morphology as well as DNA (https://www.wi
ldlifeforensicscience.org/become-certified/).
Challenges for morphological analysis
All analytical methods have limitations. Such limitations must be
borne in mind in all scientific work, and especially in the forensic
context, as the US legal system demands certainty “beyond a reasonable
doubt.”
Unsuitable evidence
Related species usually exhibit diagnostic class characters allowing
morphological identification based on the whole organism. However,
these may be lacking in partial evidence. For example, all North
American hawks of the genus Buteo can be readily distinguished by an
experienced morphologist based on plumage, but some species cannot
be reliably separated based on skeletons. Thus, skeletal remains may be
identifiable only to genus by morphological analysis. This may still
provide sufficient information for a wildlife crime investigation, since
many organisms are protected as groups. All North American hawks are
protected under the Migratory Bird Treaty Act.
Some frequent types of wildlife evidence lack taxonomically-
diagnostic morphological characters. Blood and tissue samples are ex­
amples. Other evidence in wildlife forensics casework is sometimes too
decomposed, fragmentary, or deliberately altered to permit morpho­
logical species identification. Depending on the degree of alteration,
these may include painted or dyed mammal pelts, carved bones, small
pieces of reptile leather, ivory beads, and jewelry composed of only
small body feathers. In many cases, morphological examination can
determine a higher-level identification (e.g., to family or genus), and
species identification can be made through genetic analysis (see “Case­
work Examples” section, below).
Lack of geographic information
The geographic range of a species is one of its fundamental attri­
butes. A challenge for all analysts in wildlife forensics is that the
geographic origin of items in the wildlife trade is rarely known. For
example, a shipment of reptile skins seized in San Francisco off a flight
originating in Indonesia could represent species from anywhere in the
world. In the context of morphological analysis, identification to species
may not be possible without geographical information.
As with the example of North American hawks above, this difficulty
P.W. Trail
may be alleviated by CITES listings of entire groups. For example, all
primates (order Primates) and all owls (order Strigiformes) are at least
CITES Appendix II, and all sea turtles (family Cheloniidae) are CITES
Appendix I (https://cites.org/sites/default/files/eng/app/2020/E-Appe
ndices-2020-08-28.pdf). Morphological identification at these higher
levels is sufficient to confirm that the wildlife items belong to protected
species. Also, some jurisdictions, such as the United Kingdom, follow a
strict standard under which taxa are accorded the maximum protection
applicable. Thus, evidence that could have come from either a CITES I or
CITES II species is assumed to be CITES I unless the defendant can prove
otherwise.
There are cases, however, in which a lack of geographic information
may preclude morphological determination whether wildlife evidence
item came from a highly protected species or population. For example,
African Elephants (Loxodonta africana) are listed as CITES II in South
Africa, Namibia, Botswana, and Zimbabwe, but are strictly protected
under CITES I everywhere else. Provided that sufficient population ge­
netics data are available, DNA analysis may be able to resolve such
difficult questions.
Similarity of appearance
Some closely related species share so many morphological characters
that identification is difficult, even when geographic origin is known.
Even when such species do not belong to taxa protected at a higher
taxonomic level, they may qualify for legal status under the “similarity
of appearance” clauses in some wildlife protection laws.
For example, the U.S. Endangered Species Act states “A species may be
treated as endangered or threatened if it resembles in appearance a species
which has been listed under section 4 [the section addressing listing and
recovery of species and designation of critical habitat] and enforcement
personnel would have difficulty distinguishing between the listed and the
unlisted species; if the effect of this difficulty is an additional threat to the
listed species; and if such treatment of the unlisted species would improve
protection for the listed species. A similarity of appearance listing must be
formalized by rule.” (https://www.fws.gov/endangered/about/glossary.
html; accessed August 9, 2021).
The Convention on International Trade in Endangered Species of
Flora and Fauna (CITES) states: A species may be treated as endangered or
threatened if it resembles in appearance a species which has been listed under
section 4 and enforcement personnel would have difficulty distinguishing
between the listed and the unlisted species; if the effect of this difficulty is an
additional threat to the listed species; and if such treatment of the unlisted
species would improve protection for the listed species. A similarity of
appearance listing must be formalized by rule.
(https://www.cites.org/eng/resources/terms/glossary.php;
accessed August 9, 2021).
These “similarity of appearance” provisions could greatly increase
the effectiveness of wildlife law enforcement for certain taxa, but in
practice are rarely invoked [27].
“Cryptic species”
Closely related to the issues noted above is the increased description
of “cryptic species” based on non-morphological characters such as ge­
netic and behavioral differences [28–30]. By definition, cryptic species
are difficult to distinguish morphologically. The implications of such
taxonomic decisions for wildlife law enforcement may be far-reaching,
and deserve far more consideration than they typically receive [31].
When cryptic species are granted different levels of legal protection,
DNA analysis is typically required for identification.
Despite the above challenges, morphological analysis can often
provide valuable information to inform subsequent genetic or chemical
analyses, even when it cannot achieve species identification. Examples
of synergistic morphology-genetics collaborations are presented below.
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Forensic Science International: Animals and Environments 1 (2021) 100025
Morphology and genetics: powerful partners for wildlife
forensics
Morphology and genetics are powerful and complementary tools for
the identification of evidence in wildlife crime cases. Morphology
(including microscopic examination) may be the only option for the
identification of some wildlife forensic evidence, which is too degraded,
chemically treated, or contaminated to yield viable DNA sequences for
analysis. In other cases, evidence may lack species-diagnostic morpho­
logical characters, even if morphological identification to a higher
taxonomic level is possible. For example, as described below,
morphology can confirm that a carved ivory item came from an extant
probiscidian, but DNA analysis is required to distinguish African vs.
Asian elephants (Loxodonta africana and Elephas maximus, respectively).
It is advantageous to have both approaches represented in a wildlife
forensic laboratory. Genetic analysis is able to identify many items
lacking usable morphological characters, but requires specialized
instrumentation and dedicated laboratory facilities. If these are not
available in-house, samples need to be sent to commercial labs for
amplification and sequencing, bringing significant costs and delays.
When suitable characters are present, morphological examination can
provide equally rigorous species identifications, and often obtains re­
sults more quickly and cheaply.
At NFWFL from 2010 to 2019, the Morphology Section analyzed
evidence in about 70 % more cases and identified about 85 % more
evidence items than the Genetics Section – and completed cases in less
than half the time (Table 1).
Even when morphological characters are not suitable for species ID,
morphology can often identify evidence to a higher taxonomic level,
such as family or genus. This can greatly increase the efficiency of
subsequent genetic analysis. From 2010–2019, 23 % of cases completed
by the NFWFL Genetics section also included analysis by the
Morphology section, emphasizing the importance of collaboration be­
tween these disciplines. In short, the availability of morphological
analysis allows laboratory case managers to efficiently allocate staff
time and resources, a highly useful form of triage.
A further benefit provided by morphologists is their taxonomic
expertise [32]. Unlike many DNA analysts, morphologists are specialists
in particular groups. This allows them to identify a great diversity of
species within their groups. Morphologists at NFWFL have identified
over 850 bird taxa, 100 mammal taxa, and 65 reptile and amphibian
taxa in casework. Their deep knowledge of phylogenetic relationships
enables them to provide evolutionary context and information on cur­
rent taxonomy for colleagues in other wildlife forensic disciplines.
Morphologists are also able to provide provisional identifications of
wildlife items based on emailed digital photos [33]. As described below,
this is an extremely valuable service for the FWS Office of Law
Enforcement, avoiding seizures of legal wildlife items while providing
probable cause for seizure and thorough investigation of possibly illegal
items. Finally, morphologists are able to provide training and identifi­
cation resources tailored to the needs of wildlife law enforcement offi­
cers, thereby enhancing detection rates on the “front lines” (e.g.
[34–38]).
Casework examples
The National Fish and Wildlife Forensic Laboratory employs
morphological analysts with expertise in ornithology, mammalogy, and
herpetology. From 2010–2019, bird evidence accounted for 47 % of
morphological examination requests, mammals for 42 %, and reptiles
and amphibians for 11 %. Below, we give casework examples from each
group.
Ivory
The identification of ivory to species source is a frequent request in
P.W. Trail Forensic Science International: Animals and Environments 1 (2021) 100025

Table 1
Comparison of Morphology vs. Genetics casework at NFWFL, 2010-2019.
MORPHOLOGY 2010 2011 2012 2013 2014 2015 2016 2017 2018 2019 Tot/Avg

Requests Completed 243 241 212 215 201 195 158 144 111 96 1816
Items Analyzed 1275 1718 906 981 1277 976 1015 867 1066 988 11,069
Ave Turnaround (Days) 31 46 32 27 50 69 46 50 46 32 42.9

GENETICS 2010 2011 2012 2013 2014 2015 2016 2017 2018 2019 Tot/Avg

Requests Completed 76 124 104 103 88 124 114 117 117 97 1064
Items Analyzed 278 463 793 472 271 310 559 1335 818 679 5978
Ave Turnaround (Days) 148 90 108 73 49 71 102 114 84 61 90.0

The bold values in the final column (Tot/Avg) are simply to emphasize that these are summary data.

wildlife crime investigations. Ivory evidence items are commonly Feathers

carved, modifying the appearance of the material. The varied possible
sources include not only African and Asian elephants, but also hippo­
potamus, warthog, walrus, narwhal, and sperm whale, as well as extinct
mammoth. In addition, carved bone and various synthetic materials are
often used as ivory substitutes, and these must be recognized [34].
At NFWFL, suspected ivory items are assigned first to the
Morphology Section. The initial step is to rule out synthetic ivory sub­
stitutes by illumination with an ultraviolet light source. This is followed
by close visual examination, often including magnification, to reveal
characters of bone (e.g., Haversian canals) as opposed to ivory.
Once the evidence is verified as ivory (i.e. tooth material), the ma­
terial is examined for taxonomically informative class characters. Ivory
originating from most species can be identified morphologically based
on tooth shape (if preserved in the evidence), the structure of the dentine
and cementum, and the presence or absence of intersecting lines
(Schreger lines) in the dentine [34].
Morphological examination is not, however, able to distinguish be­
tween the ivory originating from African vs. Asian elephants (Loxodonta
africana and Elephas maximus, respectively). The two species are pro­
tected differently under both CITES and US federal laws, and thus
identification to the species level is important for law enforcement.
Owing to advances in DNA amplification and analysis, it is now
routinely possible to extract DNA from ivory. Therefore, once morpho­
logical examination at NFWFL has verified that ivory evidence origi­
nated from an extant proboscidean (i.e., African or Asian elephant), it is
assigned to the Genetics Section for species ID. DNA analysis may also be
required for small or highly modified ivory items that lack informative
morphological characters.
This collaborative approach allows efficient and cost-effective
forensic processing of ivory evidence. Similar “screening” can be done
for objects suspected of being rhinoceros horn. Morphology is often able
to identify such items as synthetics or composed of bovine keratin, thus
reserving DNA analysis for those that may actually be of rhinoceros
origin. The availability of experts to visually screen evidence is a major
benefit of incorporating morphology into wildlife forensics.
Bushmeat
Bushmeat is another evidence type for which morphological exam­
ination is a useful first step, even if genetic analysis is required for
species identification. Bushmeat items received at NFWFL are usually
dried or smoked partial carcasses. The most common taxa represented
are ungulates and primates. In some cases, only meat is present, and
DNA analysis is required. Often some bones are included, however, and
morphology can distinguish general groups (e.g. monkey vs. antelope),
allowing more efficient selection of primers for DNA amplification.
Owing to the broad legal protections afforded primates, morphological
identification to that group is often all that is required, eliminating the
need for further analysis.
Feathers are the evidence items most frequently submitted to the
Morphology section at NFWFL. This is due in part to birds’ high levels of
legal protection. CITES Appendices I and II list 1434 bird species,
compared to 848 mammals and 1072 reptiles and amphibians (http
s://www.cites.org/eng/disc/species.php). An array of national and
regional laws provide strong protections for birds, including the
Migratory Bird Treaty Act (MBTA) in the United States, the Wildlife and
Countryside Act in the United Kingdom, and the Birds Directive of the
European Union. In addition, state laws and regulations protect many
bird species not listed at the federal or international level, such as
gamebirds.
Birds are particularly suited for morphological identification. They
are highly visual and often exhibit distinctive plumage patterns for
species recognition and courtship. Thanks to the long tradition of public
as well as scientific interest in birds, there are unrivalled identification
resources available. Most major families are the subjects of morpho­
logical treatises providing information on diagnostic characters, intra­
specific variability, and measurement data (e.g., [39–47]). In recent
years, the proliferation of curated image databases has provided
photographic documentation of virtually every species of bird in the
world (e.g., Birds of the World, https://birdsoftheworld.org/bow/h
ome), as well as their feathers (The Feather Atlas of North American
Birds, https://www.fws.gov/lab/featheratlas/) and skulls (https://skull
site.com/).
As with ivory and rhinoceros items, collaboration among analytical
sections is valuable for the examination of bird evidence. The Pathology
Section of the Laboratory often receives bird remains for cause-of-death
examination. Species identification of these remains is routinely con­
ducted by Laboratory ornithologists. The Morphology and Genetics
sections work together on cases in which it is important to determine the
minimum number of individuals (MNI) represented in a large assem­
blage of remains. MNI can be estimated morphologically, based on
numbers of discrete body parts (e.g., heads, feet, or wings) or of
assignable feathers (generally limited to tail feathers and outer pri­
maries). A more complete accounting of number of individuals can be
provided by DNA analysis. This depends on the availability of a
comprehensive population genetic database for the species in question,
to provide baseline data on variation. If such data are available, genetic
analysis can greatly increase the accuracy of MNI estimation. In one
complex case with multiple evidence seizures, morphological analysis
provided an MNI estimate of 9 Golden Eagles, whereas subsequent DNA
analysis confirmed the presence of 22 Golden Eagles in the evidence.
Reptile leather
The reptile evidence most frequently submitted to NFWFL is leather
from a wide variety of groups, including crocodilians, snakes, lizards,
and sea turtles. Tanned leather is a challenge for both morphological and
genetic analysis. Leather items are often composed of only pieces of the
skin, and they may be dyed or otherwise altered, making morphological
species ID difficult. The tanning process also destroys much DNA.

5
P.W. Trail
Increased sensitivity in instrumentation and analysis now allows many
tanned leather items to be identified genetically, but this is a time-
consuming process. Fortunately, many reptile taxa found in the wild­
life trade are protected at the group level. For example, all species of sea
turtles and wild crocodilians are CITES I, and all pythons and boas are
either CITES I or II. Therefore, morphological examination is often
sufficient to establish that a protected reptile taxon is represented in
evidence, even if species ID is not possible.
Provisional IDs
A final important service provided by morphologists at NFWFL is to
make provisional identifications from images of wildlife products or live
animals [33]. Enforcement personnel working at airports and other
ports of entry are tasked with monitoring legal trade in animal products
and intercepting illegal trade. These inspectors often must make rapid
evaluations of wildlife items, especially those in the possession of pas­
sengers passing through Customs. It also frequently happens that
smuggled live animals are seized. These cannot be submitted to forensic
laboratories for direct examination, both because of quarantine re­
quirements and because crime labs lack facilities to maintain live ani­
mals. Photographs of such objects and animals are emailed to NFWFL
morphologists for a quick provisional identification, which can usually
be returned the same day.
NFWFL has adopted a protocol for laboratory morphologists to
provide these provisional photo-IDs. Because a complete analysis of
morphological characters is not possible from photographs, these iden­
tifications are accompanied by the following disclaimer: “Important
Notice: This is a provisional identification/evaluation based on the
image(s) provided. This does not constitute a forensic analysis, and a
forensic report will not follow. If a forensic analysis and formal exami­
nation report are required for legal purposes, please submit the item(s)
to the Laboratory for direct examination.”
The ability of expert morphologists to provide photo-IDs to
enforcement staff in the field greatly increases the efficient handling of
wildlife items. These provisional identifications can provide probable
cause to detain suspect items, while allowing legal animal products to
pass. Common examples of such legal items are jewelry with chicken
feathers and objects made from cattle horn. In the case of live animals,
analysts may be required to travel to quarantine facilities to confirm
provisional IDs with direct examination, while other suspect items can
be submitted to the laboratory for full forensic analysis [33].
From 2010–2019, NFWFL morphologists responded to more than
3300 photo-ID requests and provided provisional IDs on over 17,600
photographs submitted by enforcement officers. These requests excee­
ded the number of cases actually received at NFWFL during this time
period (Table 1).
Conclusions
Morphological descriptions define the diagnostic characters of every
taxonomic group, and are the basis for the recognition of new species.
Examination of morphological characters is the standard method for
identification of wildlife and wildlife remains in most areas of biology,
including biodiversity survey work, ecology, and paleontology. Wider
use of morphology would provide many benefits for wildlife forensics,
including increased speed and lowered costs of species identification in
the laboratory; front-line training of enforcement staff; and verification
of reference specimens. It would make possible productive collabora­
tions with DNA analysts, increasing the efficiency of their work. Finally,
it would allow provisional identifications to be made from photographs,
providing otherwise unavailable expertise to officers in the field. The
adoption of standards and guidelines for morphology by the Society for
Wildlife Forensic Science indicates the integration of this field in the
wider discipline, and its importance for the future of wildlife forensic
biology.
6
Forensic Science International: Animals and Environments 1 (2021) 100025
Disclaimer
The findings and conclusions in this article are those of the author
and do not necessarily represent the views of the U.S. Fish and Wildlife
Service.
Declaration of Competing Interest
The authors report no declarations of interest.
Acknowledgements
I thank Barry Baker and Mary Burnham-Curtis for thoughtful reviews
of the manuscript, and Brad Foster for tabulating casework data. My
colleagues at the National Fish and Wildlife Forensics Laboratory,
especially Edgard Espinoza, Mary Burnham-Curtis, and fellow mor­
phologists Barry Baker, Ariel Gaffney, Rachel Jacobs, Cookie Sims, and
Bonnie Yates, all provided invaluable insights and support.
References
[1] C. Gans, Vertebrate morphology: tale of a phoenix, Amer. Zool. 25 (1985)
689–694.
[2] R.L. Lyman, Assumptions and protocol of the taxonomic identification of faunal
remains in zooarchaeology: a North American perspective, J. Archaeological
Method and Theory 26 (2019) 1376–1438, https://doi.org/10.1007/s10816-019-
09414-0.
[3] F. Sornoza-Molina, J.F. Freile, J. Nilsson, N. Krabbe, E. Bonaccorso, A striking,
critically endangered, new species of hillstar (Trochilidae: Oreotrochilus) from the
southwestern Andes of Ecuador, Auk 135 (2018) 1146–1171.
[4] K.F. Liem, D.B. Wake, Morphology: current approaches and concepts, in:
M. Hildebrand, D.M. Bramble, K.F. Liem, D.B. Wake (Eds.), Functional Vertebrate
Morphology, Harvard University Press, Cambridge, Mass, 1985, pp. 336–377.
[5] G.V. Lauder, Functional morphology and systematics: studying functional patterns
in an historical context, Ann. Rev. Ecol. Syst. 21 (1990) 317–340.
[6] D. Gifford-Gonzalez, Bones are not enough: analogues, knowledge, and interpretive
strategies in zooarchaeology, J. Anthropological Archaeol. 10 (1991) 215–254,
https://doi.org/10.1016/0278-4165(91)90014-O.
[7] M. Hildebrand, G. Goslow, Analysis of Vertebrate Structure, 5th ed., Wiley, 1998.
[8] K.F. Liem, W.F. Walker Jr, W.E. Bemis, L. Grande, Functional Anatomy of the
Vertebrates: an Evolutionary Perspective, 3rd ed., Harcourt College Publishers,
2001.
[9] M.A. Ashley-Ross, G.B. Gillis, A brief history of vertebrate functional morphology,
Integr. Comp. Biol. 42 (2002) 183–189.
[10] R.W. Scotland, R.G. Olmstead, J.R. Bennett, Phylogeny reconstruction: the role of
morphology, Syst. Biol. 52 (2003) 539–548.
[11] F. Watson, Descriptive taxonomy: the foundation of biodiversity research, in: C.H.
C. Lyal, C.A. Pendry (Eds.), The Systematics Association Special Volume 84,
Cambridge University Press, 2015.
[12] K.V. Kardong, Vertebrates: Comparative Anatomy, Function, Evolution, 8th ed.,
McGraw-Hill Education, New York, NY, 2018.
[13] M.K. Burnham-Curtis, P.W. Trail, R. Kagan, M.K. Moore, Wildlife forensics: an
overview and update for the prosecutor, United States Attorneys’ Bull. 63 (2015)
53–69.
[14] SWGFAST, Scientific Working Group on Friction Ridge Analysis, Study, and
Technology, Document # 103: Individualization/Identification Position Statement,
2012. http://clpex.com/swgfast/Comments-Positions/130106-Individualizatio
n-ID-Position-Statement.pdf.
[15] NAS, Strengthening Forensic Science in the United States: A Path Forward, 2009.
https://www.nap.edu/catalog/12589/strengthening-forensic-science-in-the-unite
d-states-a-path-forward.
[16] C. Neumann, I.W. Evett, J. Skerrett, Quantifying the weight of evidence from a
forensic fingerprint comparison: a new paradigm, J. R. Stat. Soc. Ser. A 175 (2012)
371–415.
[17] B.T. Ulery, R.A. Hicklin, J. Buscaglia, M.A. Roberts, Repeatability and
reproducibility of decisions by latent fingerprint examiners, PLoS One 7 (2012)
1–12. https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.
0032800&type=printable.
[18] M.J. Kelly, Computer-aided photograph matching in studies using individual
identification: an example from Serengeti cheetahs, J. Mammal. 2001 (82) (2001)
440–449. https://academic.oup.com/jmammal/article/82/2/440/2373037.
[19] L. Hiby, P. Lovell, N. Patil, N.S. Kumar, A.M. Gopalaswamy, K.U. Karnath, A tiger
cannot change its stripes: using a three-dimensional model to match images of
living tigers and tiger skins, Biol. Lett. 5 (2009) 383–386, https://doi.org/10.1098/
rsbl.2009.0028.
[20] D.T. Bolger, T.A. Morrison, B. Vance, D. Lee, H. Farid, A computer-assisted system
for photographic mark–recapture analysis, Methods Ecol. Evol. 3 (2012) 813–822,
https://doi.org/10.1111/j.2041-210X.2012.00212.x.
[21] E. Katherine Moore, et al., The Society for Wildlife Forensic Science standards and
guidelines, Forensic Science International: Animals and Environments 1 (Advances
P.W. Trail
in Wildlife Forensics) (2021), https://doi.org/10.1016/j.fsiae.2021.100015. In this
issue.
[22] J.E. Winston, Describing Species: Practical Taxonomic Procedure for Biologists,
Columbia University Press, New York, 1999.
[23] Q.D. Wheeler, Taxonomic triage and the poverty of phylogeny, Phil. Trans. R. Soc.
Lond. (2004) B359571–359583, https://doi.org/10.1098/rstb.2003.1452.
[24] M.C. Ebach, C. Holdredge, More taxonomy, not DNA barcoding, Bio. Sci. 55 (2005)
822–824.
[25] Agnarsson, Kutner, Taxonomy in a changing world: seeking solutions for a science
in crisis, Syst. Biol. 56 (2007) (2007) 531–539, https://doi.org/10.1080/
10635150701424546.
[26] P. Audisio, Insect taxonomy, biodiversity research and the new taxonomic
impediments, Fragm. Entomol. 49 (2) (2017) 121–124. http://www.
fragmentaentomol.org/index.php/fragmenta/article/view/252/256).
[27] E.O. Espinoza, J.L. Espinoza, P.W. Trail, B.W. Baker, The future of wildlife forensic
science. Pp. 343-358, in: J.E. Huffman, J.R. Wallace (Eds.), Wildlife Forensics:
Methods and Applications, Wiley, 2012.
[28] B.L. Stuart, A.G.J. Rhodin, L.L. Grismer, T. Hansel, Scientific description can
imperil species, Science 312 (2006) 1137.
[29] W.C. Funk, M. Caminer, S.R. Ron, High levels of cryptic species diversity
uncovered in Amazonian frogs, Proc. Royal Soc. B 279 (2011) 1806–1814, https://
doi.org/10.1098/rspb.2011.1653.
[30] P. Pulido-Santacruz, A. Aleixo, J.T. Weir, Morphologically cryptic Amazonian bird
species pairs exhibit strong postzygotic reproductive isolation, Proc. R. Soc.B.
(2018), https://doi.org/10.1098/rspb.2017.2081.
[31] R.L. Jacobs, B.W. Baker, The species dilemma and its potential impact on enforcing
wildlife trade laws, Evol. Anthropol. 2018 (2018) 1–6.
[32] J.E. Winston, Twenty-first century biological nomenclature – the enduring power
of names, Integr. Comp. Biol. 58 (2018) 1122–1131.
[33] P.W. Trail, Morphological species identifications of wildlife forensic evidence
based on digital images. Forensic Science International, Forensic Science
International: Animals and Environments 1 (2021) 1, https://doi.org/10.1016/j.
fsiae.2021.100021.
[34] B.W. Baker, R.L. Jacobs, M.-J. Mann, E.O. Espinoza, G. Grein, in: C. Allan (Ed.),
CITES Identification Guide for Ivory and Ivory Substitutes, 4th ed., World Wildlife
Forensic Science International: Animals and Environments 1 (2021) 100025
Fund Inc., Washington, DC, 2020. Commissioned by CITES Secretariat, Geneva,
Switzerland.
[35] P.W. Trail, Identification Guide to the Decorative Feathers of Pheasants, Chickens,
Turkeys, and Related Birds. Identification Guides for Wildlife Law Enforcement No.
12, USFWS, National Fish and Wildlife Forensic Laboratory, Ashland, OR, 2012.
https://www.fws.gov/lab/idnotes/_PheasantFeathers_final.pdf.
[36] P.W. Trail, Identification of Bald and Golden Eagle Feathers. Identification Guides
for Wildlife Law Enforcement No. 15, USFWS, National Fish and Wildlife Forensic
Laboratory, Ashland, OR, 2014. https://www.fws.gov/lab/idnotes/EagleID_
fromScans_final_small.pdf.
[37] P.W. Trail, Identifying bald versus golden eagle bones: a primer for wildlife
biologists and law enforcement officers, J. Fish Wildl. Manag. 8 (2017) 596–610.
[38] P.W. Trail, A.M. Gaffney, Distinguishing Gamebird and Waterfowl Feathers from
Raptor Feathers. Forensic Science International: Animals and Environments Vol. 1,
Published online at:, 2021 https://www.sciencedirect.com/science/article/pii/
S2666937421000081.
[39] Peter. Pyle, Identification Guide to North American Birds, Part I. Bolinas, Slate
Creek Press, California, 1997.
[40] Peter. Pyle, Identification Guide to North American Birds, Part II. Point Reyes
Station, Slate Creek Press, CA, 2008.
[41] Clifford B. Frith, Bruce M. Beehler, William T. Cooper, Michael McGuire, The Birds
of Paradise, Paradisaeidae, Oxford University Press, Oxford, England, 1998.
[42] Tony Juniper, Mike Parr, Parrots: A Guide to Parrots of the World, Yale University
Press, New Haven, Connecticut and London, England, 1998.
[43] Paul A. Johnsgard, The Pheasants of the World: Biology and Natural History, 2nd
ed., Smithsonian Institution Press, Washington, D. C, 1999.
[44] James Ferguson-Lees, David A. Christie, Raptors of the World, Houghton Mifflin
Company, Boston, Massachusetts and New York, New York, 2001.
[45] Sebastien Reeber, Waterfowl of North America, Europe & Asia, an Identification
Guide, Princeton University Press, Princeton/Oxford, 2015.
[46] Jeff. Baker, Identification of European Non-Passerines; A BTO Guide, The British
Trust for Ornithology, Thetford, 2016.
[47] Lukas Jenni, Raffael Winkler, Moult and Ageing of European Passerines, Second,
Christopher Helm, Bloomsbury, London, 2020.