Whereas the postmortem examination of a human is referred to as an autopsy, the

postmortem examination of an animal is referred to as a necropsy. The necropsy is an

essential component of any quality veterinary medical practice and may be an essential
skill for field biologists and other scientists as well. I n captive animal populations, the
necropsy is an opportunity to learn from the death of an animal so that futures illnesses
in other animals may be prevented or more easily diagnosed and treated. I n wild animal
populations, the necropsy may provide vital data about population declines, habitat
changes, effects of human or other animal populations, and causes of catastrophic die-
offs. A necropsy may not provide all the answers, but it is an important first step (and
sometimes last step) in the investigation of animal and environmental health issues.
A basic rule of necropsy is that “fresh is best.” I deally, the necropsy should be
performed as soon as possible after death. Decomposition or autolysis is a common
problem in reptiles because many species are housed in heated enclosures or aquaria,
which accelerates decomposition. I f the necropsy is delayed, carcasses should be cooled
on ice or refrigerated. Smaller specimens (less than 10.0 kg) are better preserved by
refrigeration than larger animals. Under optimal refrigeration conditions, adequate
preservation for histopathological examination of most major organs is retained for
around 72 hours if the initial postmortem condition of the carcass is good. Freezing should
be avoided and is used only as a last resort to prevent severe decomposition. Freezing
renders the carcass useless for many diagnostic purposes, as histologic architecture is
distorted or destroyed and many bacterial and fungal pathogens do not survive the freeze
and thaw process.
Decomposition of fresh specimens or thawing of frozen specimens during
shipment is an important consideration. Commercially available cold packs should be
used for carcasses or samples that must remain cold but not frozen during shipment. I ce
(wet ice) should be avoided as it will melt and tends to leak, regardless of how well the
package is sealed. Cold packs should be placed along the inside of Styrofoamlined boxes
or coolers and below the carcass or sample. TO prevent freezing, do not place cold packs
directly on top of a sample and do not use dry ice. D ry ice, however, is ideal for samples
that should remain frozen during shipment. Alternatively, multiple cold packs can be used
and should be placed on top of a sample to maintain the lowest possible temperature.
 After completion of the necropsy, the carcass should be disposed of in a manner
compliant with federal, state, and local agencies. R egulations regarding carcass disposal
vary widely among states and even within states depending on location and county, thus
the appropriate authority should be consulted. Burial may or may not be an option in some
areas. Some states and counties allow carcasses to be dumped at local landfills. A nother
option is a commercial disposal service that disposes of carcasses for a fee. A lso, most
diagnostic laboratories provide incineration or disposal services as part of the necropsy
examination.

Dissection and Internal Examination

The class R eptilia includes a vast array of species with great diversity in individual
body types. Despite this diversity, the internal anatomy is fairly well conserved across all
species with the exception of some key differences. Dissection techniques will be
described for each of the major reptile body types. T he location of major organs presented
in this section can also be found in Chapter 1.
Reptiles may exhibit prolonged involuntary movement of skeletal muscle and
cardiac contractions following death, which can be unsettling to prosectors and observers.
O ther animals thought to be dead may in actuality still be alive. Determining when some
reptiles are truly dead is not as easy as it is with a mammal and bird. Large tortoises can
be especially challenging. Some prosectors choose to decapitate freshly dead or
euthanized reptiles to ensure that the animal is definitively dead prior to dissection.

Nekropsi Ular
Prior to examination of a snake, there are special precautions that must be
considered if working with a venomous species. The same precautions apply to venomous
lizard species. A s the first step, the head should be secured with the mouth closed by
taping the jaws shut or by inserting the head into a hard cylindrical container. An empty
syringe case works well to secure the head, depending on the size of the specimen being
examined. T he head should then be removed and placed into formalin to deactivate the
venom prior to collection of the brain or any other samples from the head. Most, if not
all, venom components are destroyed by formalin fixation; however, it is unclear if some
compounds may remain toxic in some instances, especially if fixation of deep tissues is
incomplete. T herefore, the heads of venomous species should always be handled with
extreme care by experienced personnel and eye protection should be used when incising
venom glands. A lso, be aware of the location of the fangs when handling both venomous
and nonvenomous species to avoid personal injury.
Dissection of the snake is begun by placing the animal in dorsal recumbency. A n
incision is made with a scalpel or scissors along the ventral midline from the cloaca
cranially to the intermandibular space. I n larger snakes, it may be necessary to make the
initial skin incision at the lateral edge of the thick ventral scales. O nce the skin incision
has been made, the skin is reflected laterally along the length of the incision to expose the
underlying subcutis and muscle. Muscle sections can be collected at this time. The
temporomandibular junction is then incised to facilitate detailed inspection of the oral
cavity. The tongue, glottis, proximal esophagus, and trachea can be examined and
collected.
The coelomic cavity is entered via a midline incision and the entire length of the
cavity is visualized at this time. Snakes in fair or good nutritional condition have
prominent fat bodies in the coelomic cavity that extend cranially to the middle region of
the body. A s in other species, the thymus and endocrine organs cranial to the heart are
identified and collected before dissection continues. The thymus in snakes is a paired
structure with cranial and caudal lobes located cranial to the heart. T he snake thyroid
gland is a single structure (Lynn, 1970). Snakes possess two pairs of parathyroid glands;
one pair is often located between the anterior and posterior lobes of the thymus, the
second pair located at the bifurcation of the carotid artery (Clark, 1970).
The coelomic viscera of the snake can often be removed in toto. T he esophagus
and trachea are transected caudal to the pharynx. With gentle caudal traction on the free
ends of the esophagus and trachea, the ceolomic viscera can be lifted caudally from the
carcass using blunt dissection techniques or occasional sharp dissection to sever
connective tissue attachments. The viscera are removed caudally to and including the
cloaca and are placed on the dissection table for examination and sampling. Be aware of
the scent glands in the cloacal region, which will emit a strong odor if punctured.
 Snakes have a three-chambered heart comprised of two atria and one ventricle. A
n incision is made through the apex and continued into the atria and major vessels to
visualize the endocardial surfaces and valves. N ext, the trachea, bronchi, and lung(s) are
examined. In many snakes, the left lung is significantly smaller than the right or
completely absent. Boids are among the species with a well-developed left lung. T he
trachea should be opened and the incision continued into the axial chamber to allow
complete examination. The snake lung contains a large axial chamber that terminates into
a long air sac that may extend into the most caudal aspects of the coelom. Multiple
samples should be collected from different areas of the lung.
The liver is an elongate brown organ. The gallbladder in most snakes is located
distal to the liver and is connected by a long common bile duct. The spleen is often located
distal to the stomach within the mesenteric connective tissues and is a small reddish round
structure. The pancreas is a smooth or multilobular, pale tan organ located caudal to the
spleen near the duodenum. Some snake species possess a fused spleen and pancreas (a.k.a.
splenopancreas). a useful technique for locating the spleen and pancreas is to first locate
the gallbladder and then examine the surrounding tissues for these organs. The
gastrointestinal tract of snakes is a simple structure comprised of the stomach, small
intestine, and large intestine. T he small and large intestines are difficult to distinguish
during postmortem exam and multiple representative sections should be collected.
The adrenal glands of snakes are thin, elongated structures located within the
connective tissues that support the gonads (mesorchium and mesovarium) in both males
and females (Gabe, 1970). The adrenal glands can often be recognized by their
yellowishtan color and are typically collected with the gonads. Snakes lack a urinary
bladder. The kidneys of snakes are multilobularand are located cranial to the cloaca. The
kidneys are usually dark brown, but will turn light tan in reproductively active males due
to sex segment formation.
At this stage of the necropsy, all that remains in the carcass is the spinal column,
associated musculature, skin, and head. If not previously sampled, various muscle groups
can be examined and collected at this time. It is a good practice to palpate the ventral
surfaces of the vertebrae of snakes for irregularities. Vertebral disease may be examined
by collecting cross sections of vertebrae whole into formalin (less than 1 cm in thickness)
for later decalcification and histologic examination. The head is removed at the
atlantooccipital junction. For small snakes (those with heads measuring less than 2 cm in
length), the head may be collected whole into formalin, later decalcified, and then serially
sectioned. For larger snakes or cases requiring detailed examination of the nervous
system, the brain should be removed with small bone-cutting shears or a Dremel tool.
There are two basic methods for removing the spinal cord. The first method
involves separating segments of the vertebral column and extracting the spinal cord from
each segment. This technique is useful for larger reptiles, including crocodilians, and is
an alternative if a Stryker saw or D remel tool is not available. The second method exposes
the spinal cord by performing a dorsal laminectomy using a Stryker saw or D remel tool
and rongeurs. This method requires some experience to perform without damaging the
spinal cord, but allows more careful examination of the spinal cord and potentially
produces less histologic artifact.
Removal of the eyes of snakes requires special consideration due to the presence
of the spectacle, which is a fused eyelid complete with a thin dermis and epidermis. It is
important that the orientation of the spectacle and cornea remain preserved, especially for
evaluation of ocular disease. The entire globe and associated spectacle are removed by
cutting a square in the periorbital skin and dissecting around the globe, severing
extraocular muscles and other attachments.

Light Microscopy
The most common preservative used for diagnostic samples collected at necropsy is 10%
neutral phosphate buffered formalin (NBF). T issues collected in formalin are used for
histopathology, special stains for infectious agents, and some molecular tests. T he two
key considerations for preserving tissues in formalin are: (1) samples must be of the
proper thickness and (2) an adequate amount of formalin must be used. T o achieve
adequate fixation, tissue samples generally must be around 1.0 cm or less in thickness.
Formalin can penetrate only 0.5 cm of tissue in 24 hours; therefore, sections that are too
thick will decompose (autolyze) despite being immersed in formalin preservative. Among
the most common reasons for improper sizes of tissue samples are inadequate cutting
instruments and cutting surfaces; therefore, be prepared with a sharp knife, scalpel, or
razor blade and have a cutting board available. The ratio of 10% N BF to tissue should be
10:1 (i.e., 10 times as much liquid as tissue present in the container). Failure to fix tissues
in the appropriate amount of formalin will similarly result in autolysis of the tissues after
collection. Very small tissues (< 5.0 mm in size) can be placed into plastic cassettes to
ensure they are not lost during submission (Figure 4.49). Hard tissues, such as bone,
should be fixed in a separate formalin container to allow adequate penetration and fixation
prior to decalcification. T he eyes from larger specimens will also require decalcification
due to the presence of scleral ossicles (bones). T he brain from cases with evidence of
neurologic disease can be specially fixed in high-concentration formalin. First, the brain
is placed whole into a separate container with enough 37% formaldehyde to just cover
the brain tissue. Next, 10% N BF is added to the container until the brain floats neutrally
buoyant. Fixation of the brain tissue is complete when the brain sinks to the bottom of the
container, usually after approximately 24 hours.

Clark, N B. 1970. T he parathyroid, in Biology of the Reptilia, Vol 3,Gans C and Parsons
T S (Eds.), A cademic Press, N ew Y ork, 235–261.
Lynn WG. 1970. T he thyroid, in Biology of the Reptilia, Vol 3, Gans C and Parsons TS
(Eds.), Academic Press, New York, 201–234.
Gabe, M. 1970. T he adrenal, in Biology of the Reptilia, Vol 3, Gans C and Parsons TS
(Eds.), Academic Press, New York, 236–318.
Jacobson, E.R,. 2007. Color Atlas and Text: Infectious Disease And Pathology Of
Reptiles. University of Florida College of Veterinary Medicine Gainesville,
Floridac ISBN 0‑8493‑2321‑5. Taylor & Francis Group, LLC.