This document provides guidelines for collecting and handling specimens for laboratory diagnosis of animal diseases. It discusses the importance of collecting sufficient samples before treatment for various diagnostic tests. Guidelines are provided for collecting samples from live animals and during necropsy, including types of samples, containers, labeling, and storage/transport. Proper personal protective equipment, supplies, and documentation are emphasized to ensure sample integrity and lab results.

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Collection and Handling of Specimens for Laboratory Diagnosis
Dr. Milagros R. Mananggit
Chief, Regional Animal Disease Diagnostic Laboratory
Department of Agriculture, Regional Field Unit 3
Purpose of collecting samples
1. Direct examination thru microscopy
 Blood parasite examination
 Fecalysis
 Impression smears for Fluorescent Antibody Test
2. Isolation of causative microorganisms
 Bacterial Isolation and identification
 Tissue culture
 Egg inoculation
 Mice Inoculation Test
3. Serological Investigation
 ELISA
 AGPT
 HA-HI Test
 Rapid plate test
4. Molecular Test
 LAMP
 Polymerase Chain Reaction (PCR)
 Conventional PCR
 Real Time PCR (quantitative PCR)
5. Diagnosis of the disease
Purpose of testing samples
1. Confirmatory diagnosis of suspect or clinical case
2. Demonstration of freedom from infection in a defined population
3. Eradication of disease or elimination of infection from defined populations
4. Estimation of prevalence of infection or exposure to facilitate risk analysis
5. Certification of freedom from infection or presence of the agent in individual animals
or their products for trade or shipment.
6. Determination of immune status of individual animals or populations
7. For specific therapeutic medication
Basics in collecting samples
1. Always protect yourself, other people and the environment
2. Use clean, preferably sterilized implements
3. Collect samples before treatment is initiated
4. Sample volume or quantity must be sufficient for all necessary tests with enough
residual specimen for archival purposes
5. Practice proper restraint of animals during sample collection
6. Packaging containers for specimens should be leak-proof

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7. Label all specimen containers and account for all of them in a sample information
sheet
8. Maintain cold chain during transport of samples
9. Provide sufficient epidemiological information on the case
10. Notify the receiving laboratory
Checklist for Field Sample Collection
1. Personnel Protective Equipment (PPE)
 Scrub suit
 Face mask
 Gloves
 Boots
 Goggles
 Apron
2. Necropsy Kit
 Knife
 Scissors
 Bone cutter
 Forceps
 Scalpel with blade
2. Sample Containers
a. Blood/Serum
1) Vacutainer tubes
 Violet or green top –whole blood
 Red top – serum samples
 Glass slides –blood smears
2) Disposable Syringes
3) Cryo-tubes for serum (2 ml)
4) Vacutainer needle and holder
b. Organs/Tissues – each organ must have separate container
 Screw cap sterile containers
 Zip lock plastic bags
c. Swabs - tracheal, oro-pharyngeal, cloacal with viral or bacterial transport
media
4. Labelling of Samples
 Permanent water proof pen
 Masking tape
5. Preserving samples
 Ice chest/Styropor box
 Gel coolant

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 10 % buffered formalin for histopathology
 Slide box
 Methanol to fix blood smears
6. Recording Forms
 Surveillance or disease investigation forms
 Sample submission forms
8. Restrainer – rope, nose grip, pig restrainer
9. Disinfectant (hand sprayer)
 70 % alcohol
 10 % lysol or bleaching agent
10. Documentation –camera, tablet, cell phone
10. Garbage bag
Sample Collection during Necropsy
Tissue samples should be collected aseptically using rat tooth forceps and scalpel blade
dip in a container with alcohol. Cut tissue samples with demarcation between normal and
abnormal with enough quantity to perform differential diagnosis. Each tissue must have a
separate sterile container. Intestines should be tied with at both ends.
The following table show the samples that should always be collected during necropsy:

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Sample Collection in Live Animals
1. Blood
 jugular vein - horses, and small ruminants
 tail vein - big ruminants,
 wing vein - avian species
 lateral saphenous or middle cephalic in dog/cat
Serum: Let stand at room temperature for 12 to 24 hours then transfer into
cryotubes
BPE: Use violet or green top cryotubes, blood smears can be prepared, air
Dry and fix in methanol for 3 minutes.
2. Swabs – collect exudates from abscess and lesions, oro-pharyngeal, tracheal and
cloacal swabs for avian influenza
3. Feces – take directly from rectum or just after defecation
4. Urine - collection thru catherization
5. External parasites
 large parasites can be picked off and place in container with 70% alcohol for
identification
 smaller parasites such as mites can be scraped with blade and placed in slide
with mineral oil
6. Fluid from joints
7. Milk – 5 to 10 ml in sterile container
Sample Collection for Toxicology
1. Body fluids –blood and urine
2. Tissues to collect at necropsy – stomach or rumen contents, feces, brain, liver,
kidney, body fat, skin, and urine
3. Suspected source – feeds and water, poisonous plants, soil
4. Test for nitrates – best sample is aqueous fluid from the eye of dead animal
Sample Collection for Specific Diseases
Abortion
 mid portion of kidney, placenta, adrenal gland, liver, spleen, tied-off stomach,
brain and serum
 Submit under refrigeration
Abscesses
 Purulent exudates on a sterile swab or in a sterile tube collected from the margin
of the abscesses

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 Submit under refrigeration
Arthritis
 Swab or fluid from affected joint or unopened joint
 Send in bacterial transport medium, fluid in syringe
Anthrax
 cotton swab soaked in exuded blood taken from superficial ear vein or cut
surface of hemorrhagic lymph node placed in tube media.
 Submit under refrigeration
Blackleg and Malignant Edema
 2 inches cube of affected muscle
 Packed in sterile container and submit under refrigeration
Brucellosis
 Blood and serum sample collected 10 to 20 days after abortion, aborted fetus,
tied off stomach
 Shipped in sealed container under refrigeration
Caseous Lymphadenitis
 affected nymph nodes
 place in sealed container and submit under refrigeration, preserve 2x2 cm in
buffered formalin
Colibacillosis
 package section of small intestine, mesenteric lymph node, spleen, liver, kidney
 ship under refrigeration
Haemophilus
 large portion of the lung or pleural fluid from untreated sick animal with
respiratory infection
 Keep under refrigeration
Leptospirosis
 blood collected at time of clinical signs then 10 to 20 days after,
 submit under refrigeration
Mastitis
 Milk, 5 to 10 ml in sterile container
 Keep under refrigeration
Pasteurellosis
 3 inches square of affected lung and mediastinal lymph node, deep nasal or
tracheal swab
 Keep under refrigeration

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Salmonellosis
 Sections of liver, spleen, kidney and tied off section of intestine
 Submit under refrigeration
Avian Pest
 Brain, lung, trachea, spleen
 serum collected on the 4th and 14th days following onset of the disease
 Freeze and submit under refrigeration
Pseudorabies
 Brain, tonsil, lung, lymph node and serum
 Freeze and submit under refrigeration
Rabies
 Unopened head of the animal
 Freeze and send under refrigeration, don’t preserve
Swine Flu
 Lung, trachea, mediastinal lymph node
 Freeze and submit under refrigeration
Aflatoxicosis
 Suspected feed sample
 Keep dry and cool
Tumor
 Portion of tumor mass and adjacent normal tissue
 Preserve in 10% buffered formalin
Write the Report
No necropsy is complete until all findings have been recorded in written form. The report
should include the following information:
1. Owner’s name, address and contact number
2. Description of the animal: species, breed, age, sex
3. Duration and condition of the outbreak
4. Mortality and morbidity rate
5. Animal population and nearby farms
6. Clinical signs observed
7. Treatment given and vaccination
8. Feeds and water given
9. Possible contact with other animals
10. Tentative diagnosis
11. Necropsy report
12. Samples collected and submitted
13. Veterinarian’s name, address and contact number

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Packing and Transport of Samples
1. Keep tissue samples cool to prevent decomposition and growth of nonspecific bacteria
2. Keep various tissues separated from one another using ziplock or whirlpack bags, or
sterile screw cap containers.
3. Swabs should be kept moist by using sterile saline/water or transport media and keep at
4 degrees Centigrade till reach the lab.
4. Blood samples should be kept at room temperature for 12 to 24 hours before separating
the serum to avoid hemolysis. Freeze if can’t reach the lab for a week.
5. Fecal samples should be kept cool but don’t freeze.
6. Samples for bacteriology should be kept under 2-4 degrees Centigrade. Those for
virology can be frozen.
Key Concepts in Sending Specimens to the Laboratory
1. Speed in getting the materials to the laboratory.
2. Keep the samples cool on the way to the laboratory.
3. Use packaging that will prevent leakage and crushing.
4. Be sure that all your samples are well-labeled.
5. Be sure that appropriate paperwork is included with all of the samples such as necropsy
report, sample submission form, disease investigation form.
6. Alert the laboratory on the arrival of your samples.
Storage and Archives
 Future research and development efforts
 Retrospective studies
 Epidemiological studies
 Providing critical reference materials used in assay standardization, validation and
proficiency testing programs
 System of documentation and storage conditions
 Biosecurity measures
 Back-up plan in case of compromises to the storage environment
References
− Brown, Corrie, et al. A Field Manual for Collection of Specimens to enhance Diagnosis of
Animal Diseases. University of Georgia Athens Diagnostic Laboratory.
− PCAARRD. Field Sample Collection: A Quick Reference for Swine Respiratory Disease
Investigation and Diagnosis.
− BAI. Animal Health Personnel Field Manual. Quezon City
− Manual of Diagnostic Tests and Vaccines for Terrestrial Animals,
http//www.oie.int/manual-of-diagnostic-test-and-vaccines-for-terrestial-animals/