Illustrated Guide to Poultry Necropsy and Diagnosis
1. Illustrated Guide to
Poultry Necropsy and Diagnosis
Authors:
Benjamin Lucio-Martinez, DVM, MS, PhD
Jodi A. Korich, DVM
Photography, Graphic Design, and Layout:
Scott A. Birch
Project Coordination:
Jose J. Bruzual DVM, MAM, MS, DACPV
Morella De Rosa DVM, MS, DACPV
Anatomy Illustrations:
Illustrated by Kip Carter, MS CMI
Cover Design and Shipping Illustrations:
Suzanne Kabat
Content Review:
Alfonso Torres, DVM, MS, PhD
Abraham J. Bezuidenhout, BVSc, DVSc
Cesar A. Sandoval, DVM, PhD
Belinda S. Thompson, DVM
Disease Images:
Cornell University College of Veterinary Medicine Collection
Benjamin Lucio-Martinez
Bension Perelman
Plum Island Animal Diagnostic Center
Copyediting:
Ithaca Content Architecture and Design
United States Department of Agriculture, Foreign Agricultural Service
Cornell University College of Veterinary Medicine
Ithaca, New York
Credits 1
VetBooks.ir
6. Right kidney Gizzard (ventriculus)
Fig. 5a Ventral thoracoabdominal cavity
(heart and liver removed), immature female or male
Pancreas
Duodenum
Tendon
Proventriculus
Gizzard
(ventriculus)
Fig. 5b Proventriculus and gizzard (section)
6 Clinical Anatomy
•
VetBooks.ir
7. Sacral plexus
(dorsal to kidney)
Remnant of right
oviduct
Vent
Costa! nerve
- ....,*-Al-+- Lung (cut)
,...____:;;.Jllp,....'-1-1---- Spleen
..,,,,;~i.-+:l+-- Left adrenal gland
g /di-:t-;1
0
Fig. 6a Ventral thoracoabdominal cavity ( /4,f',t,fz
(GI tract removed), immature female
Bursa of Fabricius
~ - - - - - Left adrenal gland
--.....- /
Ductus deferens
Fig. 6b Immature male
.______Coprodeum
Fig. 7 Bursa of Fabricius (sagittal section)
Clinical Anatomy 7
VetBooks.ir
8. Choana
Glottis
Primary bronchus
{
Costal impression
Crop
Gall bladder
8
lnfraorbital sinus
Lung
Cranial
renal artery--+---'--4..,,::...- ..u,.....1,
External
iliac artery ---1-- .....,.....- JIIW!Pl
Sacral plexus
External
ischiatic
artery
Lung
Anterior lobe
"""""'f'--- of kidney
Middle lobe
of kidney
Posterior lobe
of kidney
Ureter
Anterior thoracic
air sac opening
Fig. 8 Urinary system
Abdominal air sac
opening
Posterior thoracic
air sac opening
Fig. 9 Respiratory system
Ceca
Rectum
Bursa of
Fabricius
diverticulum
Cloaca ~
Vent
Fig. 10 Digestive system
Clinical Anatomy
VetBooks.ir
9. Qapter 2: Field Supplies and Preparation
INTRODUCTION
SELECTING A LOCATION FOR THE NECROPSY
When selecting a location to perform the necropsy
examination, it is very important to consider bio-
security. Avoid entering the farm unless absolutely
necessary, especially when an infectious problem is
suspected. Ideally, the necropsy should be performed
in a well-lighted location outside the farm.
BIOSECURITY PRINCIPLES
1. Wear appropriate attire
When it is necessary to enter the farm, take biosecu-
rity precautions. Wear appropriate protective attire
to prevent transferring infectious agents from one
farm to another.
2. Don't bring infectious agents into the farm
Always assume that the vehicle in which you arrive is
"dirty" with respect to the farm you are entering.
3. Don't bring infectious agents into your vehicle
Always assume that the farm is "dirty" with respect
to your vehicle.
Field Supplies and Preparation
TIPS FOR SELECTING ATTIRE
From a biosecurity standpoint, two very important
pieces of attire are coveralls and foot protection.
Plasticized coveralls (Tyvek™) provide excellent
protection because they are water-proof and pro-
tect from dust, dirt, blood, feces, water, and other
liquids. However, plasticized coveralls can be very
uncomfortable in hot and humid weather. Wearing a
long-sleeved shirt underneath the plasticized
coveralls is more comfortable and facilitates getting
in and out of the coveralls in hot weather.
Cloth coveralls provide an alternative to plasticized
coveralls; however, they must be disinfected be-
tween uses. To prevent the cross-contamination of
clothing in the washer, used coveralls must be
disinfected prior to washing.
Sturdy, over-shoe rubber boots should be worn over
your shoes. Disposable boots tear easily, leaving the
shoes underneath exposed to dirt and water.
Disposable plastic boot covers worn under the rub-
ber boots provide an extra layer of biosecurity while
walking around the vehicle, and facilitate putting on
and taking off the rubber boots.
9
VetBooks.ir
12. PREPARATIONS
The primary goal of personal protective equipment is not only to protect the examiner, but
also to ensure that no pathogens are introduced or removed from the poultry operation.
To protect your footwear from farm con-
tamination, put on disposable plastic boot
covers before you step out of the vehicle.
In addition to protecting your shoes,
disposable plastic boot covers also make it
easier to slip into your coveralls.
Step out of your vehicle and finish
getting into your coveralls.
Put over-shoe rubber boots on over the
plastic boot covers.
Put on a disposable head cover and
gloves, ensuring that the sleeves of the
coverall stay inside the glove.
You are now ready to perform the
necropsy examination of the birds.
See page 48 for disrobing protocol.
1?
The best quality histological
samples are obtained by fixing
tissues in 10%formalin, buffered
to a pH of 7.
Follow the formula (shown at
right) to create a formalin solu-
tion suitable for tissue fixation.
Formalin (40% formaldehyde solution) 100 milliliters (ml)
Sodium phosphate, monobasic, monohydrate 4 grams (g)
Sodium phosphate, dibasic, anhydrous 6.5 grams (g)
Add double distilled Hp to a total volume of 1000 milliliters (ml)
The terms formalin and formaldehyde are sometimes incorrectly used interchangeably. In fact, formalin is the
aqueous solution of formaldehyde, which is a gas in its natural state. The most concentrated commercial forma-
lin solutions contain 37 to 40%formaldehyde in water. Therefore, a 10%formalin solution contains 3.7 to 4.0%
formaldehyde.
~it:til rl C11nnl ioc ::a,nrl Dr.on::. r::titil"n
•
VetBooks.ir
13. Chapter 3: Necropsy and Diagnostic Sample Collection
PREPARATION
Wet the body to keep dander and feathers away from the examination field.
Add a surfactant (quaternary ammonium,
detergent, or soap) to the water at the
concentration stated on the label.
EXAMINING THE HEAD
Dip the body of the chicken from the
neck down in the solution.
•
•
•
microbiology
(fresh tissue)
histopathology
(10%formalin)
microbiology
(swab sample)
The icons used throughout this
guide indicate the types of
diagnostic samples that can be
collected during the necropsy
exam.
A good place to begin your examination is with the external structures of the head. Following a
systematic order each time you perform a necropsy exam will help to ensure that nothing is missed.
EYELIDS
Examine the eyelids for inflammation and exudate.
Necropsy and Diagnostic Sample Collection 13
VetBooks.ir
14. Some changes in the eye are characteristic of certain diseases
and will aid in the diagnosis.
The comb is a highly vascularized tissue, and it reflects the health and hormonal
status of the chicken. Look for edema, petechiae, cyanosis, necrosis,
papules, scabs, and ulcers.
14
The cornea, anterior chamber, and lens are
clear. The iris is uniform in shape and color.
The eyelids are free of crusts, inflamma-
tion, and without exudate under the third
eyelids.
The size and shape of the comb varies with
the breed, sex, and sexual development.
Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
15. .,
BEAK AND NOSTRILS
Examine the beak for proper trimming, scabs, and nostril exudate.
INFRAORBITAL SINUSES
The infraorbital sinuses are air-filled cavities on each side of the head. Sinusitis would
result in swelling and nasal discharge.
Necropsy and Diagnostic Sample Collection
Beak color varies by breed, diet, and stage
of egg production.
---~-- -- -
Beak is symmetrical with the upper and lower
halves forming a smooth line of articulation.
Beak may be trimmed to reduce cannibalism.
The nostrils should be slightly moist and clean.
When viewed from the front, each sinus
should be slightly depressed or flush with
the side of the head.
There should not be discharge from the
nostrils when pressure is applied to the
sinus.
15
VetBooks.ir
16. EXAMINING THE ORAL CAVITY
The oral cavity should be examined closely. Its mucosa might have ulcers, erosions, opacities, and
diphtheritic membranes associated with a variety of diseases. The cleft on the roof of the oropharynx
(choana) opens into the nasal cavity and is a good place to collect samples for respiratory disease
diagnosis.
Structures of the Head
16
The surface is pink and uniform.
A row of well-defined papillae are
located near the base.
Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
17. .,
Expert Tips: Swab samples
Swab samples might not contain as much infectious material as sections of fresh tissue. However, swabs
offer a very convenient method to ship samples to a diagnostic laboratory for microbiological testing. Keep
in mind that there are different types of swabs on the market, and those intended for use in virus isolation
contain antibiotics, which should not be used for samples destined for bacterial isolation.
Necropsy and Diagnostic Sample Collection
BU BB ·- Culureswab 'E
Collection and Transport System
17
VetBooks.ir
18. EXAMINING THE BRAIN
Exposing the brain for gross examination and diagnostic sample collection is important if nervous signs are
observed clinically.
Remove the comb.
Starting at the foramen magnum, cut top of
skull around the brain with shallow snips.
s-----....----
Examine the surface of the brain
and cerebellum in situ.
Peel the skin laterally
to fully expose the skull top.
Lift and remove the skull top.
I
Separate the brain from the cranial
nerves and spinal cord.
Place the brain on a sterile surface for
inspection.
*Keep brain separate from other tissues to prevent neutralization of virus by antibodies that may be present in other tissues.
18 Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
19. .,
* Bouin's solution is recommended over formalin for fixing the eyeball.
Expert Tips: Collecting high-quality diagnostic samples
• To avoid damaging tissues destined for histological examination, it is important to use sharp scissors or a knife
when collecting tissue samples.
• To ensure proper fixation, trim tissue sections to a size no greater than 5 mm thick.
e Immerse tissue sections in 10 times their volume of 10% buffered formalin for 24 hours (see page 12 for further
instructions on preparing 10% formalin).
• After 24 hours, formalin-fixed tissues can be transferred to a single screw top jar (containing just enough 10%
buffered formalin to keep them wet) and submitted to the laboratory.
• Bouin's solution is preferred over formalin for fixing soft and delicate structures, such as the eye.
• For bacteriological examination, swab samples should be obtained as aseptically as possible. Stab a sterile surface
of the organ with a swab or sear the surface with a hot spatula before stabbing it with a swab or bacteriological
loop.
• Fresh tissue samples destined for microbiological examination should be submitted in sterile containers.
• Depending on the tissues involved and the type of isolation desired, you may wish to keep fresh tissue samples
separated from one another by placing them in different containers. Keep in mind that, for bacteriological exami-
nation, it is preferable to keep septic tissues (i.e., respiratory and digestive tract) separated from aseptic samples
(i.e., liver, spleen, heart, and ovary).
• For virus isolation, most fresh tissue samples can be pooled and submitted in one container. Tissues that should
be submitted individually include the brain, as well as any tissue with heavy bacterial contamination. It is im-
portant to keep brain tissue out of contact with other tissues, because antibodies that might be present in other
organs can neutralize low numbers of viruses present in the brain.
Necropsy and Diagnostic Sample Collection 19
VetBooks.ir
20. REMOVING THE SKIN
For the next stage of the exam, you will begin by removing the skin from the breast region.
20
Expand the cut between the legs
and the abdomen.
Lift and cut the strip of skin over the
abdomen.
Dislocate the legs to maintain the body on
its back.
Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
21. .,
EXAMINING THE BREAST
Inspect the breast region for lesions.
Peel the skin toward the
entrance of the chest to
expose the breast.
OPENING THE NECK
Assess body condition, judging by muscle mass.
Assess the symmetry of the keel,
looking for deviations (depicted). Examine the
ribs and rib joints for collapse and enlargement.
Look for excess fat, ascites, and exudate before
cutting the abdominal wall.
Expose the neck tissues and examine the trachea, esophagus, nerves, thymus, and crop.
Separate the skin of the neck from the neck tissues,
alternating between the use of fingers and the use of scissors.
Necropsy and Diagnostic Sample Collection
Fully expose the underlying tissues
of the neck.
21
VetBooks.ir
22. EXAMINING STRUCTURES OF THE NECK
22
Trachea
The crop is a diverticulum of the distal esoph-
agus. Might be distended with gas, water, or
food (as depicted).
Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
23. .,
EXAMINING STRUCTURES OF THE NECK
Necropsy and Diagnostic Sample Collection
Nerves
Vessels
jugular
vein
The jugular vein runs along both
sides of the neck.
Thymus
The thymus consists of 4 to 7 lobes parallel
to the vagus nerve. It atrophies at sexual
maturit or due to disease.
23
VetBooks.ir
25. .,
OPENING THE THORACOABDOMINAL CAVITY
In the next stage of the exam, you will carefully lift the breast to examine the air sacs, then remove the
breast and examine the viscera in situ.
IIMake a small cut through the abdominal wall
at the tip of the keel.
Place thumb under the edge of the keel and lift carefully to
avoid ripping the air sacs.
Examine the thoracic and abdominal air sacs. These membranes are thin and transparent,
and should be free of foam, fibrin, veins, fluid, or exudate.
Make a longitudinal cut above the rib joints. Repeat on the other side and completely remove the breast.
Necropsy and Diagnostic Sample Collection 25
VetBooks.ir
26. 26
Expert Tips: Maintaining knives and scissors
• Keep your necropsy kit well stocked with sharp scissors and knives.
• During the necropsy, it is important to maintain sharp edges on your knives. Always
carry a knife sharpener in the field.
• Because it is difficult to sharpen scissors in the field, use your knife as much as
possible and only use your scissors for tissues such as the trachea, esophagus, and
intestine.
• Avoid cutting through feathers because they will quickly dull the blades of knives
and scissors.
• If a sterile knife or scissors are required, you can sterilize them in the field using
10% formalin.
a) First, remove any organic residue from the instruments.
b) Wearing gloves, moisten a gauze pad with 10%formalin and wipe the surface of the
knife or scissors. Remember to do this in a well-ventilated area.
c) Before using the instruments, allow the formalin to evaporate completely until the
instruments are dry.
Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
27. .,
IN SITU INSPECTION OF THE HEART
In the next stage of the exam, you will make a visual inspection of the organs of the thoracoabdominal
cavity in situ, beginning with the heart. The heart is positioned just below the thoracic inlet.
Look for lesions on the
epicardial surface or surrounding fat.
Necropsy and Diagnostic Sample Collection
Atria may be filled with blood a nd appear dis-
tended. This photo also shows petechiation, an
artifact resulting from CO2 euthanasia.
27
VetBooks.ir
28. IN SITU INSPECTION OF THE LIVER
The liver is located in the middle of the thoraco-abdominal cavity. It has a right and left lobe attached at
the base. The left lobe is smaller than the right. Keep in mind that its size is variable and the liver may
extend beyond the edge of the sternum.
28
/
Look for gross lesions, such as areas of hemorrhage or
necrosis, abnormal coloration, and hepatomegaly.
···-·"'
Normal color (shown here) ranges from reddish
brown t o slightly yellow, depending on amount of
fat infiltration.
Swelli ng or cirrhosis might be associated wi th
t hickening of the liver edges.
-------
Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
29. .,
IN SITU INSPECTION OF THE GIZZARD
The gizzard (ventriculus) is located below the liver. It is the grinding portion of the stomach and has four
distinct bands of smooth muscles, arranged in a circle and anchored centrally by tendons.
Assess the muscle walls for changes in color,
thinning or laxity, or other gross lesions.
Necropsy and Diagnostic Sample Collection
Size varies with diet, whole grains and grit
result in a larger gizzard than a milled-grains
diet.
29
VetBooks.ir
32. REMOVING AND EXAMINING THE HEART
32
Elevate the heart and cut the great
vessels to remove the organ.
Insert scissors into the left lumen
and open it.
Examine the relative size of the right and left
Cut the apex of the heart. ventricular lumens. The left lumen is larger
than the slit-like right lumen.
Inspect the surface of the myocardium Repeat on the right side and inspect
and valves. the myocardium and valves.
Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
33. .,
EXAMINING THE SYRINX AND BRONCHI
The distal end of the trachea narrows to form the syrinx,
then bifurcates into the primary bronchi.
Examine the lumen for exudate
or parasites. The narrow syrinx
is susceptible to obstructions.
Expert Tips: Handling of aseptic and septic tissues to ensure high-quality samples
• At this point in the exam, most of the tissues that you have collected for testing are asep-
tic. Aseptic samples have minimal bacterial contamination and are well suited for the isola-
tion of pathogenic bacteria and viruses. In most cases, it is best to collect aseptic tissues
before handling the septic tissues like the gastrointestinal tract. Once the gastrointestinal
tract is opened, large numbers of bacteria are released, which might interfere with testing.
• Keep in mind that autolysis causes the intestinal wall to deteriorate very quickly. When
histological evaluation of the intestinal wall is indicated, samples should be collected as
close to the time of death as possible, preferably in freshly euthanized birds.
• It is best to collect diagnostic samples from aseptic tissues in a timely manner. After
death, bacteria migrate from septic tissues (especially the intestine) and contaminate
tissues that are normally aseptic, confounding the diagnosis.
• When attempting to isolate viruses from septic tissues, collect the samples as soon as
possible to avoid gross contamination of these tissues with bacteria.
Necropsy and Diagnostic Sample Collection 33
VetBooks.ir
34. REMOVING THE GASTROINTESTINAL TRACT
To examine the gastrointestinal tract, begin by removing it from the abdominal cavity. In this step of the
exam, you will remove the proventriculus, gizzard, liver, and intestine as one unit, setting them aside for
later examination.
Sever the proventriculus from the
esophagus.
Cut the attachments anchoring the pro- Gently pull and linearize the intestine, being
ventriculus, gizzard, liver, and intestine. careful not to tear and leak its contents.
EXAMINING THE BURSA OF FABRICIUS
The bursa of Fabricius is found on the dorsal wall of the cloaca. Pull back the intestine to expose it for
examination. The bursa can also be exposed via a dorsal approach through the skin.
I
34
The integrity of the bursa is a good indicator of
immune competence in sexually immature birds.
It will completely regress in sexually mature chickens.
In the absence of immunosupressive infections,
the bursa reaches its maximum size around 8
weeks of age.
Necropsy and Diagnostic Sample Collection
a
VetBooks.ir
35. •
Separate the bursa from the cloaca. Open the bursa to expose the lumen.
Examine the pink folds of its mucosa. Look
for hemorrhage, edema, or inflammation.
Sever the posterior end of the intestine from the cloaca. Lay the GI tract aside for examination after the
remaining aseptic tissues have been processed.
Necropsy and Diagnostic Sample Collection 35
VetBooks.ir
36. EXAMINING THE LUNGS
Now that the GI tract is out of the way, perform a more thorough examination of the deeper tissues,
starting with the lungs.
Use the edge of the scissors to bluntly dissect the lungs from the rib cage.
36
Lungs are pinkish red and slightly spongy
on palpation. Look for focal or regional
abnormalities.
Post-mortem congestion (depicted)
results in reddening and should not be
mistaken for disease.
Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
37. •
EXAMINING THE BRACHIAL AND INTERCOSTAL NERVES
With the lungs removed from the rib cage, the brachia[ plexus and intercostal nerves may be examined.
lntercostal nerves are thin and white.
The intercostal nerves run parallel to the ribs.
Expert Tips: Necropsy and sampling recommendations
• During the necropsy, clean your scissors often to avoid transferring blood and other material between
tissues.
• When cutting through skin, place the pointed tip of the scissors up and the blunt tip down
(ie., between the skin and the subcutaneous tissue).
• To open organs with a small lumen, place the pointed tip inside the lumen.
• Samples accidentally contaminated with bile are not suitable for bacteriological diagnosis, but may
still be submitted for histopathological or virological examination.
Necropsy and Diagnostic Sample Collection 37
VetBooks.ir
38. EXAMINING THE GONAD AND ADRENAL GLANDS
The ovary should be examined more thoroughly at this time. In female chickens, only the left
reproductive tract develops. The ovary is located posterior to the lungs and anterior to the kidneys.
The size of the ovary varies greatly with
age and sexual status.
In sexually immature females, ovaries
are small structures with a fine nodular
appearance.
An active ovary has follicles in various
stages of development.
EXAMINING THE INFUNDIBULUM, TESTES, AND ADRENAL GLANDS
In hens in lay, the infundibulum is a large structure associated with the follicles. In males, the bilateral
testes are anterior to the kidneys.
Examine the infundibulum and serosal
surface of the oviduct.
38
Depending on the sexual development, the testes vary greatly in size. The adrenal
glands are small, yellowish tan glands located anterior to the kidneys. They are
partially obstructed from view by the testes (male) and ovary (female).
Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
39. •
EXAMINING THE KIDNEYS
At this stage, the kidneys are fully exposed, facilitating a more thorough examination.
The kidneys are composed of three distinct lobes on each side. The lobes are tightly adhered within the
bony synsacrum of the pelvis.
Look for hemorrhage, swelling, nodules,
and excessive amounts of white urates.
Necropsy and Diagnostic Sample Collection
the fossa of the synsacrum.
39
VetBooks.ir
40. EXAMINING THE SACRAL PLEXUS AND SCIATIC NERVE
Once the kidneys have been assessed, they are removed to expose the underlying sacral plexus and
vertebra for examination.
Using your fingers, gently scrape the kidneys out
of the synsacrum to expose the sacral plexus.
Nerves are pale yellow to white and smooth.
Follow the plexus through the
abdominal wall, where it emerges as
the sciatic nerve in the leg.
40
Remove a section of the sciatic
nerve from the leg.
Repeat on the other leg and compare sides.
Diameter of each nerve should be similar
and fine striations might be visible.
Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
41. •
* Submission methods for nerves vary, with some labs preferring nerve tissue submitted on a flat surface (like a tongue
depressor) to prevent contraction artifacts. Other labs prefer nerve samples to be submitted in a coil to facilitate
cutting and examination.
EXAMINING THE RIBS AND VERTEBRAE
The ribs, rib joints, and vertebrae are all now fully exposed for examination. Abnormalities in these bony
structures can be associated with nutritional imbalances or infections.
Look for collapsed ribs or enlargements of the
joints (costochondral junctions).
Necropsy and Diagnostic Sample Collection
Look for vertebral column deviations,
swellings, or fractures.
41
VetBooks.ir
42. EXPOSING THE ANTERIOR GASTROINTESTINAL TRACT
With the aseptic tissue examination and sampling completed, you can now focus your attention on the
septic tissues, beginning with the caudal oropharynx, esophagus, and crop.
Cut through the commissure
of the beak.
Inspect the oropharynx for diphtheritic Continue cutting down the length of the
membranes, hemorrhages, nodules, or ulcers. esophagus and crop.
EXAMINING THE ANTERIOR GASTROINTESTINAL TRACT
Inspect the mucosal surfaces of the esophagus and crop.
42
Wash away food particles
and evaluate the mucosa.
The mucosa[ surface of the esophagus is pink
and glistening, with glands and longitudinal
folds.
Look for proliferative lesions, ulcers,
and erosions of the crop.
Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
43. •
EXAMINING THE PROVENTRICULUS AND GIZZARD
In the last stage of the necropsy, you will focus on the gastrointestinal tract (GIT). The GIT is usually
reserved for last to avoid contaminating other organs with intestinal content. However, if GI disease is
suspected, examine the GIT as soon as possible to minimize the effects of autolysis on the intestine.
The GIT exam begins with an inspection of the serosal surfaces.
Necropsy and Diagnostic Sample Collection
Look for lesions involving the
honeycomb-shaped glands
(visible through serosal wall).
Palpate wall for flaccidity or dilation.
43
VetBooks.ir
44. EXAMINING THE INTESTINAL SEROSA
When examining the intestine, look for regional abnormalities like proliferation or
dilations, as well as any specific focal lesions such as hemorrhages or necrosis. Keep in
mind that the anatomical sections of intestine of the chicken are not well demarcated, and
are often referred to as the anterior, middle, and posterior thirds.
44 Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
45. .,
Anaerobic Intestinal Sample Collection
EXAMINING THE PROVENTRICULUS AND GIZZARD MUCOSA
Using scissors, open the gastrointestinal tract to expose the mucosal surfaces for inspection.
There are a variety of mucosal lesions to look for, including hemorrhages, necrosis, ulcers,
proliferations, and nodules.
Necropsy and Diagnostic Sample Collection 45
VetBooks.ir
46. EXAMINING THE GI TRACT MUCOSA
Inspect the intestinal mucosa. If necessary, gently remove the ingesta, but do not scrape the
mucosa because this will damage the tissue and interfere with histopathology.
Open the duodenum. Assess the mucosa. Open the jejunum. Assess the mucosa.
Open the ileum. Assess the mucosa. Open the cecum. Assess the mucosa.
At base of the ceca, pay close attention to the cecal tonsils.
Normal cecal tonsils are pale pink.
46
These lymphoid structures might become enlarged,
hemorrhagic or necrotic during infection (as depicted).
Necropsy and Diagnostic Sample Collection •
VetBooks.ir
47. .,
CLEAN-UP AND DISPOSAL
Proper disposal of the carcasses is important to maintain biosecurity. If the farmer has a
biosecure method for disposal, have the carcasses disposed of at the farm. If not,
remove them for sanitary disposal.
If there is a need to remove the
carcasses, place them and any dirty
supplies in a sturdy plastic bag.
Necropsy and Diagnostic Sample Collection
Place the bag in a second bag that has not touched the ground.
Seal the second bag and place it in your vehicle.
47
VetBooks.ir
48. PERSONAL PROTECTIVE EQUIPMENT (PPE) REMOVAL
It is important to remove and bag your personal protective equipment before leaving the
poultry operation to prevent the spread of disease.
Prepare a new plastic bag.
CONCLUSION
Remove your coveralls, headcover, rubber
boots, and lastly your gloves. Place them
in the plastic bag. Do not remove the dis-
posable plastic boot covers until you enter
your vehicle.
Enter your vehicle, remove your
plastic boot covers, and place them
in another plastic bag.
Prior to departing the poultry facility, fresh tissue and swab samples should be placed in a
cooler with ice packs for transport. Back in your facility, these samples will be prepared for
shipping to the diagnostic laboratory. Boots and necropsy tools should be scrubbed clean with
a detergent and soaked in a disinfectant solution (as shown below). Coveralls should be
sterilized and laundered, and your field necropsy kit re-packed for future use.
48 Necropsy and Diagnostic Sample Collection
•
VetBooks.ir
49. Chapter 4: Packaging and Shipping Diagnostic Sampl
INTRODUCTION
WHAT CONSTITUTES A PATIENT
SPECIMEN?Diagnostic patient specimens might contain poten-
tially infectious agents. To ensure public safety,
international regulations for the transport of infec-
tious materials, including dangerous goods, by any
mode of transport have been established. These
regulations include detailed protocols for the proper
packaging and shipping of diagnostic specimens to
diagnostic laboratories. To be qualified to ship dan-
gerous goods, shippers must enroll in an IATA (Inter-
national Air Transportation Association) Dangerous
Goods training course and pass a certification exam.
Regulations may change over time and shippers must
be re-certified every two years.
"Patient specimen means human or animal mate-
rial collected directly from humans or animals and
transported for research, diagnosis, investigational
activities, or disease treatment or prevention.
Patient specimen includes excreta, secreta, blood
and its components, tissue and tissue swabs, body
parts, and specimens in transport media (e.g.,
swabs, culture media, and blood culture bottles)."
WHAT CATEGORY OF SPECIMEN DO I
HAVE?
The information provided within this chapter is
intended for field reference only and is not a substi-
tute for the IATA training course. The following sum-
maries were prepared based on regulations in place
effective June, 2010, available from the
International Air Transportation Association.
The first step in preparing patient specimens for
shipment to a diagnostic laboratory is to determine
which category of specimen you need to ship. There
are three categories of specimens defined below.
Category A Substances
An infectious substance
which is transported in a
form that, when exposure to
it occurs, is capable of caus-
ing permanent disability,
life-threatening or fatal
disease in otherwise healthy
humans or animals. For a list
of infectious substances
included in Category A, see
page 50.
Category B Substances
A Category Bsubstance is an
infectious substance which
does not meet the criteria for
inclusion in Category A. Most
veterinary diagnostic speci-
mens fall within this category.
These samples do not meet
the definition of dangerous
goods.
Exempt Patient Specimens
Patient specimens for which
there is minimal likelihood that
pathogens are present are not
subject to these regulations if
the specimen is packed in a
packing which will prevent any
leakage and is marked with the
words "Exempt animal speci-
men."
Once you have identified the category of patient specimens you are shipping, refer to
the appropriate section for instructions on the proper packaging and shipping methods
for your specimens.
PackaQinQand Shiooing Diagnostic Samples 49
VetBooks.ir
50. PREPARING DIAGNOSTIC SPECIMENS FOR SHIPMENT
Category A Substances
UN2814, UN2900
Category A shipments require special-rated triple packaging, labeling, and documentation. Because these
samples are considered dangerous goods, the shipper must have received training and certification in an
IATA Dangerous Goods course. If you are suspicious of infection with a Category A pathogen in animals,
it is appropriate to notify either the local or state public health department, USDA Area Veterinarian in
Charge (AVIC), or state veterinarian who can provide assistance with sample packaging and shipping. Note
that many carriers, including the UPS and the US Post Office, will not accept these packages.
Most veterinarians will not have a need to routinely ship Category A substances.
Category A Substances - updated 2009
UN2814
Infectious substances affecting humans
and animals
Bacillus anthracis (cultures only)
Brucella abortus (cultures only)
Brucella melitensis (cultures only)
Brucella suis (cultures only)
Burkholderia mallei-Pseudomonas mallei-Glanders
(cultures only)
Burkholderia pseudomallei-Pseudomonas pseudomallei
(cultures only)
Chlamydia psittad-avian strains (cultures only)
C/ostridium botulinum (cultures only)
Coccidioides immitis (cultures only)
Coxiella burnetti (cultures only)
Crimean-Congo hemorrhagic fever virus
Dengue virus (cultures only)
Eastern equine encephalitis virus (cultures only)
Escherichia coli, verotoxigenic (cultures only)
Ebola viru
Flexal virus
Francisella tularensis (cultures only)
Guanarito virus
Hantaan virus
Hantaviruses causing hemorrhagic fever with renal syndrome
Hendra virus
Hepatitis B virus (cultures only)
Herpes B virus (cultures only)
Human immunodeficiency virus (cultures only)
Highly pathogenic avian influenza virus (cultures only)
Japanese Encephalitis virus (cultures only)
Junin virus
Kyasanur Forest disease virus
Lassa virus
Machupo virus
Marburg virus
Monkey pox virus
Mycobacterium tuberculosis (cultures only)
Nipah virus
Omsk hemorrhagic fever virus
Poliovirus (cultures only)
Rabies virus (cultures only)
Rickettsia prowazekii (cultures only)
Rickettsia rickettsii (cultures only)
Rift Valley fever virus (cultures only)
Russian spring-summer encephalitis virus (cultures only)
Sabia virus
Shigella dysenteriae type I (cultures only)
Tick-borne encephalitis virus (cultures only)
Variola virus
Venezuelan equine encephalitis virus (cultures only)
West Nile virus (cultures only)
Yellow fever virus (cultures only)
Yersinia pestis (cultures only)
UN2900
Infectious substances affecting animals only
African swine fever virus (cultures only)
Avian paramyxovirus Type 1-Velogenic Newcastle
disease virus (cultures only)
Classical swine fever virus (cultures only)
Foot and mouth disease virus (cultures only)
Lumpy skin disease virus (cultures only)
Mycop/asma mycoides- Contagious bovine pleura-
pneumonia (cultures only)
Peste des petits ruminants virus (cultures only)
Rinderpest virus (cultures only)
Sheep pox virus (cultures only)
Goat pox virus (cultures only)
Swine vesicular disease virus (cultures only)
Vesicular stomatitis virus (cultures only)
Infectious substances shown in blue are capable of causing disease in birds.
Pr1rkr1oino r1nrl <;h innino nir1onnc:tir <;;:imnli:>c: •
VetBooks.ir
51. .,
PREPARING DIAGNOSTIC SPECIMENS FOR SHIPMENT
Category B Substances
UN3373
•
0
Submission
Form
-------
f)
FORMALIN
•
0 e
.C.'i:..
e
8
0
0
f)
0
G
0
Assemble specimens.
When shipping histologic samples, fix tissues in a
10:1 ratio of 10% formalin for 24 hours. Then trans-
fer tissues to a leak-proof, screw-top jar with only
enough formalin to cover tissues. When shipping
microbiologic and histologic samples together in one
package, leave the tissue samples in the 10:1 ratio
of formalin to allow them to fix in transit.
Label all specimens with the tissue source, date,
and relevant farm/flock information.
Place jars containing 10% formalin in an approved
secondary container, separate from samples intend-
ed for microbiological analysis. Protect fragile items
(like glass) with padding. Add absorbent material
capable of absorbing the entire liquid contents.
Place all secondary containers in an approved leak-
proof package capable of protecting the contents.
If shipping by air transport, use containers rated to
an internal pressure of 95 kPa to withstand aircraft
cargohold pressure changes.
When shipping fresh tissues and/or swab samples,
place a frozen ice pack (NOT dry ice) in the shipping
container. Ice packs are unnecessary when shipping
histologic samples only.
Air cargo shipment limits:
· Primary containers cannot exceed 1 liter or 4 kg (solids).
- Entire package cannot exceed 4 liters or 4 kg total.
f) Put submission forms inside sealed plastic bag
and place inside shipping container.
e Seal shipping container and label with sender
and recipient's name, address, and phone number.
f) Affix package with a "Biological Substance, Category B.
UN3373" diamond label. Cover labels with clear tape.
4Ii) For international shipments, contact laboratory for a
copy of an import permit and instructions for properly
packaging to clear customs. Arrange all international
shipments in advance with the receiving laboratory.
Ship package, preferably by overnight delivery.
- To ship large body parts, organs, or whole bodies exceeding these limits, seek a Special Provision A82. On the waybill accompanying the
shipment, note: "Special Provision A82 (Title 49 CFR 172.102) or A81 (IATA) to exceed volume and weight limit. The quantity limits do
not apply to animal body parts, whole organs or whole bodies known to contain or suspected of containing an infectious substance, UN
3373, Biological Substance, Category B."
Packaging and Shipping Diagnostic Samples 51
VetBooks.ir
52. PREPARING DIAGNOSTIC SPECIMENS FOR SHIPMENT
Exempt Patient Specimens
•
0
Submission
Form
_*_____
f)
0 Assemble specimens.
f) When shipping histologic samples, fix tissues in a
10:1 ratio of 10% formalin for 24 hours. Then trans-
fer tissues to a leak-proof, screw-top jar with only
enough formalin to cover tissues. When shipping
microbiologic and histologic samples together in one
package, leave the tissue samples in the 10:1 ratio
of formalin to allow them to fix in transit.
0
0
Label all specimens with the tissue source, date,
and relevant farm/flock information.
Place jars containing 10% formalin in an approved
secondary container, separate from samples intend-
ed for microbiological analysis. Protect fragile items
(like glass) with padding. Add absorbent material
capable of absorbing the entire liquid contents.
Place all secondary containers in an approved leak-
proof package capable of protecting the contents.
If shipping by air transport, use containers rated to
an internal pressure of 95 kPa to withstand aircraft
cargohold pressure changes.
Air cargo shipment limits:
- Primary containers cannot exceed 1 liter or 4 kg (solids).
- Entire package cannot exceed 4 liters or 4 kg total.
0
e
0
•
0
0
... 4
~
1'!!8!MUN
,/
I
When shipping fresh tissues and/or swab samples, place
a frozen ice pack (NOT dry ice) in the shipping contain-
er. Ice packs are unnecessary when shipping histologic
samples only.
Put submission forms inside sealed plastic bag
and place inside shipping container.
Seal shipping container and label with sender
and recipient's name, address, and phone number.
Label package with "Exempt Animal Specimen."
Cover labels with clear tape.
CI!) Ship parcel, preferably by overnight delivery.
- To ship large body parts, organs, or whole bodies exceeding these limits, seek a Special Provision A82. On the waybill accompanying the
shipment, note: "Special Provision A82 (Title 49 CFR 172.102) or A81 (IATA) to exceed volume and weight limit. The quantity limits do
not apply to animal body parts, whole organs or whole bodies known to contain or suspected of containing an infectious substance, UN
3373, Biological Substance, Category B."
VetBooks.ir
53. Chapter 5: Selected Lesions and Associated Diseases
INTRODUCTION
A variety of gross lesions might be visually identified during a necropsy examination. This guide depicts a
selection of important lesions that can be seen in poultry and some of the diseases associated with them.
We have selected the most common types of lesions encountered in practice, as well as those lesions that
are associated with highly pathogenic diseases requiring rapid diagnosis for successful control. When ap-
propriate, it is important to seek expert advice from other colleagues or professionals who have expertise
in the diagnosis and management of poultry diseases.
For convenience, this guide is organized by tissue, lesion, and diseases that might be associated with
them. Because of their importance, highly pathogenic diseases are presented at the top of the differential
list, followed by the prevalent diseases. Keep in mind that disease prevalence varies by region and type of
poultry management practice (i.e., breeders, egg layers, broilers, backyard, or village poultry).
Skin erythema
• Velogenic viscerotropic Newcastle disease
• Highly pathogenic avian influenza
• Septicemia
• Alabama redleg syndrome (Marek's disease)
Plum Island Animal Diagnostic Center (PIADC)
Skin lesions
Cornell University
Swollen feather follicles
Cornell University
• Fowl pox - nodules, scabs, pustules • Marek's disease (also called skin leukosis)
• Cannibalism - scabs, blood crusts • Vesicular dermatitis
• Fowl pox
Selected Lesions and Associated Diseases 53
VetBooks.ir
65. USDA-
Jurisdictional areas
Physical boundaries
HPAI epidemiology
Infected Premises
characteristics
Contact Premises
characteristics
Environment
Climate
General area, region,
or agricultural sector
biosecurity
Number of backyard
or transitional
premises
Continuity of
business
May 2017
United States
Department of
Agriculture
Factors Used to Determine Control Area Size
• Effectiveness and efficiency of administration
• Multi-jurisdictional considerations: local, State, Tribal, and multistate
• Areas defined by geography
• Areas defined by distance between premises
• Reproductive rate
• Incubation period
• Ease of transmission
• Infectious dose
• Species susceptibility
• Number of contacts
• Transmission pathways and
transmission risk
o Extent of animal movement
oNumber of animals
oSpecies of animals
• Number and types of premises
• Susceptible animal populations
and population density
• Animal movements
• Types of premises in area or
region
• Land use in area or region
• Prevailing winds
• Modes of transmission (such as, fecal-oral, droplet,
aerosol, vectors)
• Survivability in the environment
• Ease of diagnosis (for example, no pathognomonic
signs; requires diagnostic laboratory testing)
oAge of animals
oMovement of traffic and personnel to and from
premises (fomite spread)
oBiosecurity measures in place at time of outbreak
• Movement of traffic (fomites) and personnel to and
from premises (fomite spread)
• Biosecurity measures in place prior to outbreak
• Susceptible wildlife and population density
• Wildlife as biological or mechanical vectors
• Biosecurity practices in place prior to outbreak
• Biosecurity practices implemented once outbreak detected
• Types of premises, animal movements, and network of animal and fomite movements
• Continuity of business plans and processes in place or activated at beginning of outbreak
(such as surveillance, negative diagnostic tests, premises biosecurity, and risk-
assessments)
• Permit processes, memorandums of understanding, and information management systems
in place or activated at beginning of outbreak
HPAI Response
Ready Reference Guide-Overview ofZones
Minimum Sizes of Zones and Areas
Infected Zone (IZ)
Buffer Zone (BZ)
Control Area (CA)
Surveillance Zone
(SZ)
Perimeter should be at least 3 km (-1 .86 miles)
beyond perimeters of presumptive or confirmed
Infected Premises. Will depend on disease
agent and epidemiological circumstances. This
zone may be redefined as the outbreak
continues.
Perimeter should be at least 7 km (- 4.35 miles)
beyond the perimeter of the Infected Zone.
Width is generally not less than the minimum
radius of the associated Infected Zone, but may
be much larger. This zone may be redefined as
the outbreak continues.
Perimeter should be at least 10 km (-6.21
miles) beyond the perimeter of the closest
Infected Premises. Please see the table to the
left for factors that influence the size of the
Control Area. This area may be redefined as
the outbreak continues.
Width should be at least 10 km (-6.21 miles),
but may be much larger.
For more information, please go to:
http://www.aphis.usda.gov/fadprep
For more details on zones and premises designations, please see
the APHIS FAD Framework: Response Strategies (Manual 2-0).
For the HPAI Response Plan: The Red Book, click here.
For the Secure Poultry Supply Plan, click here.
USDA APHIS Veterinary Services • National Preparedness and Incident Coordination (NPIC)
4700 River Road Unit 41 • Riverdale, MD 20737
3 of 3
VetBooks.ir
66. USDA
Infected
Premises (IP)
Contact
Premises (CP)
Suspect
Premises (SP)
At-Risk
Premises
(ARP)
Monitored
Premises
(MP)1
Free Premises
(FP)
United States
Department of
Agriculture
HPAI Response
Ready Reference Guide-Overview ofZones
Infected, Contact, and Suspect Premises are subject to individual premises quarantines.
At-Risk and Monitored Premises are subject to movement control restrictions.
Summary of Premises Designations
,@AS
Premises where a presumptive positive case or confirmed Infected Zone
positive case exists based on laboratory results,
compatible clinical signs, HPAI case definition, and
international standards. 1
Premises with susceptible animals that may have been Infected Zone,
exposed to HPAI, either directly or indirectly, including but Buffer Zone
not limited to exposure to animals, animal products,
fomites, or people from Infected Premises.
Premises under investigation due to the presence of Infected Zone,
susceptible animals reported to have clinical signs Buffer Zone,
compatible with HPAI. This is intended to be a short-term Surveillance Zone,
premises designation. Vaccination Zone
Premises that have susceptible animals, but none of Infected Zone,
those susceptible animals have clinical signs compatible Buffer Zone
with HPAI. Premises objectively demonstrates that it is
not an Infected Premises, Contact Premises, or Suspect
Premises. At-Risk Premises may seek to move
susceptible animals or products within the Control Area by
permit. Only At-Risk Premises are eligible to become
Monitored Premises.
Premises objectively demonstrates that it is not an Infected Zone,
Infected Premises, Contact Premises, or Suspect Buffer Zone
Premises. Only At-Risk Premises are eligible to become
Monitored Premises. Monitored Premises meet a set of
defined criteria in seeking to move susceptible animals or
products out of the Control Area by permit.
Premises outside of a Control Area and not a Contact or
Suspect Premises.
Surveillance Zone,
Free Area
Infected Zone (IZ)
Buffer Zone (BZ)
Control Area (CA)
Surveillance Zone
(SZ)
Free Area (FA)
Summary of Zone and Area Designations
•Zone that immediately surrounds an Infected Premises.
Zone that immediately surrounds an Infected Zone or a
Contact Premises.
Consists of an Infected Zone and a Buffer Zone.
Zone outside and along the border of a Control Area. The
Surveillance Zone is part of the Free Area.
Area not included in any Control Area. Includes the
Surveillance Zone.
Example Zones, Areas, and Premises
Premises Zones and Areas
Infected Zone • Buffer Zone • Surveillance Zone
+ • = Control Ared • + t = Free Area
Note: Figures are not to scale.
In an HPAI outbreak, the Incident Commander will work with the Operations
Section and Planning Section to determine the appropriate designations.
1 The Secure Poultry Supply sets out the "defined criteria" for Monitored Premises for this type of movement during an HPAI outbreak.
May 2017 USDA APHIS Veterinary Services • National Preparedness and Incident Coordination (NPIC)
4700 River Road Unit 41 • Riverdale, MD 20737
2 of 3
~...
~
~
t~
VetBooks.ir
67. USDA United States
Department of
Agriculture
Highly Pathogenic Avian Influenza (HPAI) Response
Ready Reference Guide-Overview ofZones
Buffer Zone Control Area
/
Unfected Zone+
Buffer Zone)
~.......-'"""'"1.........- -...- - ----
Scale
- 1.86 mil s (3 km)
- - - - - 6.2 miles {10km)
In the Buffer Zone (which 1s part
of the Control Area), there are
movement controls and
surveillance act1v1t1es
- Infected Zone • Buffer Zone • Surveillance Zone
- + • = Control Area • + f-.;-;;, = Free Area
Zone that immediately surrounds an Infected Zone or a Contact Premises.
Consists of an Infected Zone and a Buffer Zone.
Surveillance Zone (SZ) Zone outside and along the border of a Control Area. The Surveillance Zone is
part of the Free Area.
Free Area (FA) Area not included in any Control Area. Includes the Surveillance Zone.
May 2017
7'---~-
USDA APHISVeterinary Services • National Preparedness and Incident Coordination (NPIC)
4700 River Road Unit 41 • Riverdale, MD 20737
1 of 3
VetBooks.ir