Printable Version Specimen submission guidelines.pdf

Serology Specimens

  Note: for serological testing, 10-14 days are required for the development of demonstrable antibodies. If a blood sample is drawn during the acute phase of a disease, a second sample should be drawn 10-14 days later. If testing for export, subclinical carriers, pre-sale, pre-breeding examination, clinically normal animals, etc, contact the laboratory for the best samples to collect.

General Histopathology Specimens

1. Specimens should represent typical lesions, including active margins and adjacent (normal) tissue, rather than lesion cores or curetted debris. Autolysis, freezing, mutilation (forceps crushing or tearing), or removal of small samples by electrocautery will make samples unsuitable for proper evaluation.
2. Multiple specimens (from different sites or types of lesions) should be identified individually by size, suture tags, or separate containers. Samples that are < 1cm should not be sliced, but fixed and submitted for testing. Brain and eyes should be fixed whole.
3. Nearly all diagnostic histopathology can begin with tissue fixed in 10% buffered formalin.
4. The volume of fixative should be 10-20 times the specimen volume. After 12-24 hours, specimens can be transferred to just enough formalin to keep them moist during shipment. There is no need to pay for transport of excess fixative.   The tissues should be submitted overnight to the lab.
5. Wide-mouth bottles are preferred containers. Narrow-mouth bottles often have to be broken or cut to release fixed tissues. Plastic bottles are better than glass; anticipate rough handling during shipment, and package accordingly. To prevent leakage tape the lid, or wrap a piece of parafilmTM around the cap.  Label the container with animal ID and source of tissue.
6. Place the bottle in a leak proof secondary container. Place container and accession form into a mailing container. Mail to the lab as soon as possible.

Bacteriological Specimen

Used correctly, microbiologic cultures can identify etiologic agent(s) and contribute key information towards a diagnosis and treatment.  However, specimens that are not collected properly or shipped properly can lead to erroneous diagnoses.  Etiologic agents may be missed because of improper transport medium, improper transport environment, or improper preservation techniques.  We offer the following guideline to optimize these procedures for sample collection and submission.

If you have any questions, please contact Bacteriology section -phone at: 859-257-8283.

1. Samples should be collected aseptically and placed in sterile plastic bags or heat sterilized containers. Seal tightly. Do not use chemically disinfected containers, or plastic gloves or sleeves.

2.  Label all submissions with the location (tissue) and species of origin. The same bacterial species may be highly significant or a meaningless contaminant, depending on the tissue and/or species from which the sample was obtained. Also, depending on the tissue/species of origin, different culture conditions may be necessary to isolate and identify specific pathogens.

3.  Always specify the tests you want done, and the pathogens you suspect, particularly in the case of specimens with normal bacterial flora (feces, intestinal contents, skin, or oral mucus membranes). If we don't know what you're looking for, we may not inoculate the proper media to find it.

4. It is best to collect other samples before opening the gastrointestinal tract. Tissue samples (lung, liver, spleen, kidney, etc.) should be 5 g or larger to allow surface searing in the laboratory to reduce contaminants. Fecal samples should not be submitted in stoppered tubes, as fermentation will dislodge the stoppers.

5. Place each sample in a separate container. If the intestine is to be cultured, tie off both ends of a segment and place it in a separate container.

6. Except in the case of abortions, please separate samples that are to be examined by different laboratory sections. If a specimen is to be examined by both the virology and bacteriology sections, the specimen should be divided, each piece placed in a separate container, and labeled with the source of the tissue and the desired laboratory service.

7. Fluids for culture (e.g. body cavity fluids, pericardial fluid, joint aspirates) should be submitted in a sealed sterile tube, in as large a volume as is available (up to ~10 ml), since the concentration of organisms may be very low in these samples. Fetal fluids (thoracic or peritoneal fluids, or heart blood) to be examined for Leptospira sp. by FA test should be submitted in a sealed sterile tube to which 10% buffered formalin is added at a rate of 1.5 ml per 20 ml fluid. Never submit fluid in syringes, because they tend to leak in transit and contaminate packaging. Never submit fluids or other specimens for bacteriologic culture in EDTA (purple top) evacuated tubes, as EDTA is highly toxic to many bacterial species.

8.  Milk samples should be submitted in screw top tubes frozen or placed on ice packs. Less than 1 ml is required, and larger volumes are undesirable, especially if the samples are frozen. 

 9. Specimens for isolation of anaerobic pathogens require special care. Anaerobic bacteria die in the presence of oxygen and should be shipped in an anaerobic container, such as anaerobic swab or Port-a-Cul system. Cultures for Clostridium sp. in parenchymatous organs and intestines ordinarily provide no significant information concerning the cause of death if the samples are taken more than one hour after death.

10. Some specimens, such as porcine nasal swabs for Bordetella sp. isolation must be delivered to the laboratory within 12 hours of collection. Fastidious organisms such as Campylobacter spp. require special media for transport to the laboratory. Where there is any doubt as to what samples to collect and how to transport them - - CALL THE LABORATORY FIRST at 859-257-8283!

11.  When collecting large numbers of samples (e.g. >30 milk samples for mastitis diagnosis permits the laboratory to have personnel and media available for prompt processing.

12.  Keep specimens cold from the time they are collected until they arrive at the laboratory. Specimens should be shipped in insulated containers with a sufficient number of ice packs to last 48 hours. Specimens arriving in the laboratory in a decomposed state will not be processed, since processing and culture of these tissues lead to meaningless or erroneous results.
 

Parasitology

Fecal Specimens

Specimens should be submitted in individual sealed containers labeled with owner's name, animal species, and animal number/name. Whirl Pak bags, Ziplock bags (double bagged), or plastic containers are recommended.

Feces must be fresh for accurate results. Refrigerate fecal sample immediately after collection but do not freeze. Submit sample to the laboratory as soon as possible in a container with ice packs via any of the 24 to 48 hour transport services.  Do not submit in gloves.

Molecular Diagnostics

All specimens collected for molecular testing must be refrigerated (2-8ᵒC) promptly after collection and shipped on cold packs within 72 hours. If Specimens need to be held more than 72 hours, they should be stored at -70ᵒC and shipped on dry ice.

Submission form should be filled completely, and clinical history should be provided. All samples must be labelled clearly. Care must be taken to avoid cross‐contamination when taking multiple samples.

Swab samples: Samples should be collected on a synthetic swab (Dacron or Rayon) with a plastic shaft. Calcium alginate swabs are unacceptable and swabs with cotton tips and wooden shafts are not recommended. Swabs can be submitted in a transport media such as universal transport media, sterile phosphate buffered saline (PBS), or saline solution.  If a virus isolation is also requested, a viral transport medium is highly recommended.  Swabs that include additives such as agar gel-type media, charcoal and heparin should not be submitted as these additives can inhibit PCR reaction. Swabs must be kept cold and transported in cold conditions with enough ice bags.

Blood samples: Please see the web site or call the lab to obtain the preferred blood sample. The preferred blood sample could be whole blood (purple top with EDTA), serum or plasma depending on the molecular test requested.

Fecal samples:  Fecal samples from animals should be submitted in non-leak, sterile plastic container and kept cold during transport. Do not submit the fecal samples in gloves.

Tissue samples: Tissue samples from the areas of lesion should be submitted as soon as possible after death. Intestinal tissue samples should not be mixed with other type of tissues. The samples should be placed in a non-leak, sterile container and kept cold during transport.  

Contact the laboratory at 859-257-8283  if you have any other questions.

Mycology

Sample collection for evaluation for mycotic diseases. (Note: Isolation and identification of mycotic agents may require 30 days.)

1. The affected area of skin should be washed with 70% alcohol, scrapings taken from the active border area and placed in a sterile container. The basal portion of the hair or hair stubble should be plucked out with forceps and submitted as well. Hair and skin scrapings for dermatophyte isolation should be shipped dry at room temperature, between glass slides or in a paper envelope. Specimen plates and slants for fungal identification should be submitted at room temperature 
2. Exudates or tissues for culture should be collected aseptically, refrigerated, and sent to the laboratory on ice packs. It is strongly recommended that tissues be submitted for histopathology rather than mycology in the initial evaluation of these cases. Please contact the laboratory at: 859-257-8283 for additional information.

Virology

If a viral disease is suspected, it is important to collect specimens as soon as possible following onset of clinical symptoms or death of the animal. If testing for export, regulatory, subclinical carriers, pre-sale, pre-breeding examinations, etc., the timing of sample collection is also important, and the state, sale, or export agency may need to be contacted prior to collection.

Remember to keep all samples moist and cool when transporting to the lab. If shipping samples, send them by overnight transport with ice packs. Samples for virus isolation must be received within 48 hours of collection. The following are important points to keep in mind:

• Serum samples should not have any hemolysis and have an adequate amount for the lab to perform all requested tests.
• Tissues should be fresh and collected aseptically. Submit generous portions of tissue and place in sterile sealed containers or plastic bags, (i.e., Whirl-Pak bags).
• Swab samples (conjunctival, nasal, oral, fecal, vaginal, etc.) are to be kept moist with a few drops of sterile saline solution or viral transport media. Virology will not accept a swab submitted in a gel tube such as Amies. Cotton, plastic, wood-handled, and Dacron and other synthetic swabs are all acceptable.
• Submit ear notch samples individually in separate bags or tubes with clear identification.