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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.