CHAPTER 6.

Animal Specimen
INTRODUCTION
Diagnostics are important beyond the field level and the individual animal, herd, or flock. While awareness and
ability to diagnose disease helps the animals affected and the producer, it is also critical in establishment of a national
database. This database is used to make fact-based decisions for the best nation-wide control programs to finance and
support, leading to greater animal health for the national herd and flock.
Surveillance is the act of monitoring disease and taking action based on findings. It is at the core of all national
animal health programs. Central to surveillance is submission of samples to the diagnostic laboratory. Without diagnostic
activities, there is no surveillance. Without surveillance, there is no overall assessment of animal health.
Collection of animal specimens for diagnosing animal diseases is a crucial step in veterinary medicine. It allows
veterinarians to identify the causative agent of the disease and determine the best course of treatment. The specific
specimens collected will vary depending on the suspected disease, the animal species, and whether the animal is alive or
deceased.
OBJECTIVES
 Explain
Animal Specimen
Collection of animal specimen in diagnosing is done either the animals is alive or not. Following are some of the
specimen/samples that can be collected in an animal:
Blood
This is the most common sample collected from a live animal. Whole blood should be collected aseptically, typically
by venipuncture (is the process of obtaining intravenous access for the purpose of venous blood sampling or intravenous
therapy) of the live animal. The jugular vein is the preferred location for small ruminants and horses. The wing vein is the
site for birds. The tail vein is the easiest site for sample collection in cattle.. Care should be taken to collect and dispense
blood samples as gently as possible to prevent damage to red blood cells, which causes hemolysis or the destruction of red
blood cells.
Blood may be taken by syringe and needle or by needle and vacuum tube (not easy in delicate veins but convenient
in strong veins). Small quantities of blood are conveniently obtained by pricking with a triangular, solid-pointed needle.
Ideally the skin at the site of venipuncture should first be shaved (plucked) and swabbed with 70% alcohol and allowed to
dry. For samples that are collected with anticoagulant, thorough mixing, using gentle agitation only, is necessary as soon as
the sample has been taken. It may also be necessary to make a smear of fresh blood on a microscope slide; both thick and
thin smears may be prepared.
Feces
Feces can be collected freshly voided or preferably directly from the rectum/cloaca for tests such as culture for
microorganisms, parasite examination, or fecal occult blood determination; or can be collected for culture and
molecular-based diagnostics from the rectum/cloaca using cotton, dacron, or gauze-tipped swabs, dependent on
the volume of sample required by the specific test methodology. Samples collected on swabs should be kept moist
by placing them in the transport media recommended for use with the specific test to be performed, which may
range from sterile saline to culture media containing antimicrobials or stabilisers. Fecal specimens should be kept
chilled (e.g. refrigerated at 4°C or on ice)
Urine
Urinalysis provides information about the urinary system as well as other body systems. It is usually performed in-
clinic rather than being sent to an outside laboratory, which is why it is important for the veterinary team to be able to
effectively evaluate a complete urinalysis.
 In cattle, massaging the area under the escutcheon should result in a flow of urine within one minute. In sheep,
occluding the nares for a short period sometimes precipitates urination. In all other cases, catheterization is necessary
unless the animal can be confined and monitored constantly to catch the next instance of urination.
Epithelium
Epithelial tissue in the form of biopsies or skin-scrapings; swabs of oral, nasal, pharyngeal, and gastrointestinal
surfaces, as well as plucked hair or wool can be used variously for direct examinations or laboratory tests to identify
surface parasites such as mites and lice, fungal, bacterial or viral infections, allergic reactions, and neoplasia. The
specimens should be collected aseptically and preserved as specified for the intended test(s). Deep skin-scrapings
obtained using the edge of a scalpel blade are useful for burrowing mites. Feather tips have been validated for use
in the detection of viral antigen for Marek’s disease, and used as a sample for molecular detection of additional avian
diseases. Epithelial tissues, particularly those associated with vesicular lesions and collected into viral transport
media, can be critical in the laboratory diagnosis of specific viral infections such as foot and mouth disease.
Swabs are often used to collect exudates from lesions, for example taking a swab from an abscess. Additionally,
swabs are used to collect tracheal and oropharyngeal fluid samples from birds to test for avian influenza.
For smaller parasites, such as skin mites, scrape with a razor blade to be sure you go deep enough to get the
parasite. Scrape the skin with a razor blade and put the collected material onto a slide with some mineral oil. Then you can
put the slide under a microscope to see the mites.
For smaller parasites, such as skin mites, scrape with a razor blade to be sure you go deep enough to get the
parasite. Scrape the skin with a razor blade and put the collected material onto a slide with some mineral oil. Then you can
put the slide under a microscope to see the mites.
Ocular sampling
For smaller parasites, such as skin mites, scrape with a razor blade to be sure you go deep enough to get the
parasite. Scrape the skin with a razor blade and put the collected material onto a slide with some mineral oil. Then you can
put the slide under a microscope to see the mites. For smaller parasites, such as skin mites, scrape with a razor blade to be
sure you go deep enough to get the parasite. Scrape the skin with a razor blade and put the collected material onto a
slide with some mineral oil. Then you can put the slide under a microscope to see the mites.
Reproductive tract sampling
Preputial and vaginal wash fluids and swabs of the cervix and urethra can be used as specimens for investigation of
reproductive disease. The swabs should be kept moist following collection by placing in the recommended volume of
transport media required by the laboratory test, typically sterile saline or specified culture media. Semen
specimens are typically obtained using an artificial vagina or by extrusion of the penis and artificial stimulation.
Avoid contamination of the specimen with antiseptic or detergent solutions used to prepare the animal/site for
Sampling.
Milk
Milk can be collected from individual animals or from bulk milk in tanks pooled from multiple animals in a herd. The
teat(s) used for sample collection should be cleaned, and any detergent thoroughly rinsed off before collection of
the specimen. In collecting milk from individual teats, the initial stream must be discarded and only the subsequent
streams sampled. The method of preservation prior to testing varies with the requirements of the test; in some
cases it will be critical to avoid freezing or addition of chemical preservatives.
Environmental and Feed sampling
Feed - Collect a minimum of 1 liter of feed, making sure it is representative of what the animal has been consuming.
Dried hay and bales - Sample a minimum of 8 bales or piles, collecting a minimum of ten (200g each) samples. Combine
and mix samples and place a 300-350g subsample into an airtight plastic bag.
Dried grains - Collect "hand-grab" samples (100g each) from ten separate locations within the bags, piles or bins. Combine,
mix and place 300-350g subsample into an airtight plastic bag.
Fresh forages (plants) - Collect fresh plants (stems and leaves only) into a paper bag to prevent it from growing mold. Avoid
contamination by soil or animal manure.
PRESERVING SPECIMENS FOR LABORATORY USE
1. Keep the tissues cool
Bacteria are usually present in ALL collected samples. Unfortunately the nonspecific bacteria will replicate even faster
than any bacteria that might be causing a disease. So, it is important to keep the tissue cool because the bacteria grow at
warm temperatures and the nonspecific bacteria will quickly outgrow any specific bacteria we might be trying to find. Also,
the nonspecific bacteria will generate toxic products that might kill any viruses or fungus we are looking for in the sample.
2. Try to keep the various tissues separated from one another
As tissues get grouped together, bacteria from one can quickly overtake another. This is especially true if intestinal samples
or fecal materials are included. Use plastic bags that can be sealed - “ziplock” or “whirlpak” bags are the best.
3. Samples should be kept moist
If the sample dries out, any agents in there might dry out as well, making it difficult to isolate the infectious organism. For
swabs, immerse the swab in sterile saline or sterile water and keep at 4°C until it can be sent to the laboratory.
4. For serum samples – let the blood coagulate.
This is best done by taking the sample in a redtopped tube and leaving it at 37°C for 12-24 hours. The red top on
the tube is a universal symbol that the tube contains no anticoagulant; consequently the blood will coagulate, and after
coagulation, the serum can be removed and put into a clean tube. If you have a centrifuge, you can get more serum, but if
not, you should be able to get enough serum without having to spin the tube. If you don’t remove the serum, the red cells in
the clot will slowly break down, releasing their hemoglobin into the serum, causing the serum to take on a pink-red color and
making many serologic tests difficult to undertake.
Once the serum is removed and in a separate tube, keep it cool until you can get it to the laboratory. If it will be
more than one week before the sample can be shipped to the laboratory, freeze it, because antibody levels will slowly fall,
even at 4°C.
For external parasites, mites, fleas, and ticks can all be kept in 70% alcohol indefinitely.
For fecal samples, keep them cool until they can be sent to the laboratory. Nematode eggs usually survive well at
4°C but can be destroyed by freezing, so DO NOT FREEZE.
Storage and Packaging of Samples/Animal Specimen
A fundamental approach to packaging samples involves a 3-layer barrier to protect them:
 The sample is placed in an appropriate primary container (sealed jar, bag, or tube).
 The primary container is then enclosed within a secondary container, which also includes absorbent material.
 The secondary container is then placed in the shipping box (tertiary container), which often houses refrigerant or
cold packs as well as various cushioning materials (eg, polystyrene foam) to protect the sample. Ideally, the tertiary
container is a sturdy polystyrene refrigerator box or a cardboard box lined with a fitted polystyrene lining.
Items such as syringes, obstetrical gloves, and containers without sealable orifices are not suitable for shipment.
Under no circumstances should needles, scalpel blades, or any other potentially injurious item (eg, broken microscope
slides with sharp edges) be submitted to the lab.
Liquid samples should not ship in plastic bags; instead, a sealable jar should be used. Waterproof markers should
be used when labeling specimen bags and containers; at a minimum, patient identification and specimen type (eg, urine,
serum, plasma) are critical labeling information.
Coolant materials should be sealed in separate plastic bags to prevent condensation damage. Refrigerant or cold
packs should not be placed in direct contact with samples, such as tubes of whole blood, that could suffer adverse effects if
frozen in transit.
The suitably protected, completed submission form must be included.
If dry ice is used, this should be noted on the cardboard box label, and the lid should not be sealed with tape.
Otherwise, CO2 released from the dry ice could increase pressure and damage the package or contents.
Key concepts in Sending Specimen in a Laboratory
1. Speed in getting the materials to the laboratory. The longer the package is in transit, the greater probability that bacteria
will proliferate or the agent will degenerate, so that a diagnosis cannot be made. Package should be welllabeled to help
ensure that it is not delayed in transit.
2. Keep the samples cool on the way to the laboratory. Pack with cold packs to help ensure that the package does not get
warm and promote bacterial overgrowth. It is preferable not to use ice, because it will melt and may cause the package
to leak.
3. Use packaging that will prevent leakage and crushing. Leakage of contents will likely cause a package to be refused by
the shipper. Also, the package should be sturdy because if it is crushed, it is more likely that samples will be ruined or that
leakage will occur.
4. Be sure that all your samples are well-labeled. The laboratorian needs to understand what the samples are. Is the swab
from a trachea or the cloaca? Is the tissue in the bag liver or intestine? Are the serum samples from cattle or water buffalo
or chickens?
5. Be sure that appropriate paperwork is included with all of the samples. The diagnosis of disease is a group effort. Every
stop along the chain has some information to contribute. By giving the laboratorian a complete history of the problem and
any gross necropsy findings, he or she is likely to do a much better job with his/her part of solving the big puzzle. Be sure to
package the paperwork in a waterproof way so that even if some of the samples happen to leak, the paperwork will still be
readable.
6. Alert the laboratory. If you let the laboratory know that a submission is coming, testing procedures can be prepared in
time for when the sample arrives.