Necropsy Manual

Zoetis Internal Training Manual

 Table of Contents

Necropsy Manual. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Six key steps to a consistent routine

1. Obtain the History. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

A good individual animal and flock history
should be obtained.

2. Examine the Bird Externally. . . . . . . . . . . . . . . . . . . . 5

Examine the bird for any signs of trauma
and evaluate the bird’s general body condition.

3. Open the Body. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

4. Remove the Organs and

Set Aside for Examination. . . . . . . . . . . . . . . . . . . 7 – 9

5. Examine and Sample the Organs. . . . . . . . . . . 10 – 14

Lymphoid system . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Brain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
Respiratory system . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
Liver. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
Intestinal tract. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Sampling of Organs. . . . . . . . . . . . . . . . . . . . . . . . . . . 14

6. Write your Findings . . . . . . . . . . . . . . . . . . . . . . . . . . 15

Additional Resources. . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Contact information
Suggested reading

2 NECROPSY MANUAL
Necropsy Manual

Recognizing poultry abnormalities is enhanced by developing a consistent necropsy routine.

There is often a tendency to move quickly to the suspected lesion or body system, which risks
missing important information. A good necropsy involves paying attention to ALL the clues
that can be provided, so the routine has to be followed, with attention to detail at every step.

Six key steps to a consistent routine

1. O
 btain the history

2.E
 xamine the animal externally

3. Open the body

4. Remove the organs — set aside for detailed

examination and sampling

5. Examine and sample the organs

6. Write your findings

Zoetis Training Manual 3

 1. Obtain the History

A good individual animal and flock

history should be obtained. This history
should include:

1. Bird’s age

2. Sex

3. Breed

4. C
 linical signs

5. History of trauma or disease

6. History of any treatments administered

7. Any other information that may be relevant

to the case such as type of feed and water

8. If the bird is a member of a flock, the following

is also required:
•N
 umber of birds in the flock

• Number of birds in the affected group

• Number of affected birds

• Clinical signs of the flock should also be noted

A proper history can help in determining

what samples should be taken and what tests
are necessary for making the correct diagnosis.

4 NECROPSY MANUAL
2. Examine the Bird Externally

Examine the bird for any signs of

trauma and evaluate the bird’s general
body condition.
• I f possible, the bird should be weighed. Prior to opening the body, you might want to swab
•T
 he area of the keel bone (sternum) should be felt
the oropharynx, trachea or cloaca. When swabbing
the oropharynx, insert the swab up in the choanal cleft,
to determine if there is any pectoral muscle atrophy.
as depicted in Figure 3.
•T
 he skin, feathers, eyes, ears and beak should be
examined for any abnormalities. Take a close look at
the comb and wattles — any swelling or discoloration?
•L
 ook at the back of the bird for evidence
of feather picking.

Figure 3 Figure 4

Figure 4 depicts swabbing the cloaca.

Figure 1

• I f any skin lesions are noted, they should be sampled.

•E
 xamine all mucous membranes (mouth, nares,
and conjunctiva) for any discoloration or other
abnormalities. How about the cloaca? Any diarrheal
staining? Urates present? Trauma?
•L
 ook at the joints — any swelling?

Figure 5

Dip the whole carcass into a bucket of soapy water

to thoroughly wet all the feathers — this will decrease
the dander that might aerosolize from the skin and
will also keep your instruments free of small feathers
as you do the dissection.

Figure 2

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 3. Open the Body

•T
 he body should be placed on its back with its feet •E
 xtend the cut up through the cervical area and
facing you. cut open the beak at the angle of the jaw. Now oral
•R
 eflect the wings back. cavity, esophagus, trachea and crop are all visible.
•T
 he keel bone and breast muscles are then removed
•C
 ut through the skin between the legs and the breast
so the legs can be fully abducted and lie flat against by incising the pectoral muscles on each side of the
the table. keel and cutting through the ribs. Use the heavy
poultry shears.

Figure 6 Figure 7 Figure 8 Figure 9

•R
 emove the skin from the ventral surface of the bird •R
 emove the keel and breast muscles entirely — you
by cutting across at the caudal edge of the keel and should now be able to see the internal organs from
then pulling skin cranially and caudally — peel away oral cavity to rectum.
from the muscle to expose the muscular body wall. •A
 t this point, all internal viscera should be examined
•M
 ake a small cut into the body cavity using scissors in situ for any abnormalities before removing any
or a scalpel blade — just behind the breastbone — organs. Note the color, position and size of all organs
and then pull the abdominal muscle caudally and look for any adhesions (Figure 10).
to expose some abdominal viscera.

Figure 10 Figure 11

•E
 xamine the air sacs for increased thickness
or cloudiness (caudal thoracic air sac is at the end
of the forceps) (Figure 11).

6 NECROPSY MANUAL
4. Remove the Organs and Set Aside for Examination

Located in the cloaca is the light cream-colored

saccular organ, the bursa of Fabricius. The bursa
of Fabricius contains lymphoid follicles and can be
easily found in young birds. Once the bird reaches
sexual maturity, it undergoes involution and therefore
becomes smaller as the bird ages.
Cut at the end of the large intestine, but leave the
bursa in the bird.
Here are two views of the bursa — undisturbed
(Figure 13) and incised (Figure 14).

Figure 12 Figure 13

It is probably easiest to remove the abdominal viscera

first and then go back and remove the thoracic organs.
Remove the liver — in birds, the liver takes up a big
portion of the abdomen.
The spleen (Figure 12) can be a difficult organ to find
once everything is removed, so it is a good idea to
locate it now and set it aside. It is spherical in shape
and located on the right side at the junction of the
proventriculus and ventriculus. Pull the proventriculus
aside, and it should pop into view. Take it out now
and set it in a clean dry spot.
Find the junction of the esophagus and stomach, cut
here and pull the digestive tract out, all the way to the
cloaca. The digestive, urinary and reproductive tracts Figure 14
come together at the cloaca.

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 4. Remove the Organs and Set Aside for Examination

The kidneys (Figures 15-18) are nestled up against the Figure 17 is an immature female. In females, only the
body wall. There are three portions: cranial, middle and left side of the tract persists; the right is vestigial and
caudal poles. The reproductive tract lies on top of and too small to be identifiable.
at the cranial end of the kidneys. Figure 18 is a mature female.
Figure 15 is an immature male.
Figures 16 is a mature male.

Figure 15

Figure 17

Figure 16

Figure 18

8 NECROPSY MANUAL
4. Remove the Organs and Set Aside for Examination

Pull the heart and lungs away. The lungs are closely Open some joints to observe the fluid and
adhered to the dorsal body wall and, therefore, synovial membranes (Figure 21).
careful teasing of the lungs away from the ribs may be
necessary to remove them.
Open up the nasal cavity to take a close look
at the sinuses (Figure 19).

Figure 21

The brain can now be removed. Using the smaller

scissors (not poultry shears), chip away at the skull,
beginning from the foramen magnum, and remove
the calvarium covering the cerebral hemispheres and
Figure 19 the cerebellum. Take the brain out — use care, this
organ is quite soft (Figure 22).

Observe the sciatic nerve, which is an important location

where Marek’s disease can be seen (Figure 20).

Figure 20 Figure 22

Zoetis Training Manual 9

 5. Examine and Sample the Organs

It is a good idea to go from the “cleanest” organs Lymphoid system

to the “dirtiest.” Usually this order is: lymphoid tissue, The spleen in birds is a small, round organ that should
brain, lungs, heart, kidneys, reproductive tract, liver, be a uniform mahogany color on capsular and cut
intestinal tract. surfaces (Figures 24 and 25).
Note any abnormalities for each (color, consistency,
distribution and size). Be sure to examine both capsular
and cut surface. Make several cuts in each organ.
Collect specimens for further diagnostic work.

Figure 24
Figure 23

Figure 25

The bursa changes dramatically with age. Birds older

than 10 weeks will have a bursa that may even be
difficult to locate. A normal bursa in a young bird has
an accordion-like structure and is a homogeneous
tan color (refer to Figures 13 and 14).

10 NECROPSY MANUAL
5. Examine and Sample the Organs

Brain Respiratory system

Often the brain is sliced down the middle to create Begin by cutting through the larynx, trachea and syrinx,
symmetrical halves for frozen and formalin specimens. making note of any mucus, froth or petechia (Figure 29).
Lungs should be pink, “spongy” and free of any fluid
(Figure 30).

Figure 29
Figure 26

Heart
The heart can now be examined. Make one incision into
each ventricle; examine muscles and valves.

Figure 27
Figure 28

Figure 30

Kidneys
These should be smooth and homogeneous. A reticular
pattern is an indication of dehydration (Figure 31).
Figure 27 Figure 28

Figure 31

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 5. Examine and Sample the Organs

Reproductive tract Liver

Testes are homogeneous on capsular and cut The liver’s surface should be examined for any
surface (Figure 32 and 33). Ovaries should be free of abnormalities. It should be palpated for any nodules,
inflammation. Sterile egg yolk peritonitis is a common friable areas or other abnormal changes. Several slices
finding in “spent” layers. are made into the liver in order to examine the deeper
structure of the liver (Figure 34).

Figure 32 Figure 34

Figure 33

12 NECROPSY MANUAL
5. Examine and Sample the Organs

Intestinal tract
Look in the mouth for any abnormalities. Cut down
the esophagus and see how much food is in the crop
(Figure 35).

Proventriculus

Ventriculus
Figure 38
Figure 35
The ventriculus, or gizzard, should be examined
next. Because the gizzard is responsible for grinding
ingested material, it has a thick external muscularis
layer and contains small stones or grit. The ventricular
glands secrete a thick protective gel, known as koilin,
which has a yellowish color. The gizzard thickness
should be examined and the surface examined
for erosions, ulcerations, discoloration or other
abnormalities. Peel the koilin back to look at the
mucosa (Figure 39).

Figure 36 Figure 37

Open the proventriculus and make note of the

lining, which is normally bumpy due to the presence
of digestive glands. Note any abnormalities. The
proventriculus-ventriculus junction (white arrow) is
an area with abundant lymphoid tissue and should be
examined carefully for lesions (Figure 38).

Figure 39

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 5. Examine and Sample the Organs

The small intestine of birds is typically arranged into Sampling of Organs

several loops before entering the colon. The first loop
is the duodenum. It is easily identified by the location •T
 issues could be collected in duplicate, with half
of the pancreas within the duodenal loop mesentery going into 10% formalin for histopathology and
(Figure 40). half as unfixed tissues that will be used for bacterial
culture, virus isolation, fluorescent antibody testing,
toxicology, etc.
•T
 issue collection should be based on necropsy
findings and differential diagnoses.
•L
 abel the tissues that are collected fresh so that the
laboratory performing the tests can determine what
each tissue is.
Crushing should be avoided when taking samples
Figure 40
since this may cause histological artifacts. This can
be avoided by using a sharp blade and a hard surface.
The remaining loops make up the jejunum and ileum. •W
 hen examining any organ with a mucosal surface
The colon is relatively short with two long ceca and (trachea, esophagus, intestine, etc.), care should be
connects to the cloaca via the colorectum. In the taken not to damage or destroy the mucosal surface
picture below, you see the two ceca and the large colon by rubbing the surface with fingers or instruments.
in between. There are large lymphoid patches in the In order to improve fixation of tissues, samples should
proximal portions of the ceca; these are often called not exceed 5 mm in thickness, and volume of fixative
“cecal tonsils” (Figure 41). should be at least 10 times the volume of tissue.
Fresh samples should be packaged so that they
remain cool and to minimize possibilities of leaking.
Be sure to submit the proper paperwork to
Figure 41
accompany the samples.

Figure 41

Figure 42

14 NECROPSY MANUAL
6. Write your Findings

No necropsy is complete until all findings have been

recorded in written form for your records. Note that
some customers may request a necropsy report. The
report should include at least the following information:
•S
 pecies, breed, age, sex

•H
 istory

•D
 ied or euthanized?

•N
 utritional, hydration status

•F
 indings from external examination

•F
 indings by organ system: lymphoid (spleen, bursa),
respiratory, digestive, urogenital, musculoskeletal,
nervous
•D
 ifferential diagnosis based on necropsy examination

Additional Resources
For more information on this Necropsy
Manual, please contact:
Jose A. Linares, DVM, ACPV
Technical Director, Global Marketing,
Zoetis
jose.a.linares@zoetis.com

Suggested reading:
Diseases of Poultry, 12th Edition. Chapter 1, Principles
of Disease Prevention: Diagnosis and Control. pp. 33-
40. Blackwell Publishing. 2008.

A Practical Guide for Managing Risk in Poultry

Production. Published by the American Association
of Avian Pathologists, Inc. 2011.

Zoetis Training Manual 15

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