Chapter 5 - Sampling and Testing

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Annex E: Collecting and Shipping Tissues from Animals at High Risk for Tuberculosis
(Skin test reactors or those from tuberculosis infected or suspect herds)

Mycobacterium bovis can cause serious human disease. Take appropriate safety precautions when dealing with animals or tissues suspected of being infected with tuberculosis. Consult your supervisor or Safety Officer and refer to the MSDS available from the CFIA Office of Biohazard Containment and Safety, Tel: 613-225-2342 extension 4256.

1. Tissues to submit

• Perform a complete post-mortem with emphasis on incising and examining (both left and right where applicable): the following tissues and lymph nodes: Head - submandibular, parotid, medial retropharyngeal, lateral retropharyngeal, tonsils. Thorax - anterior and posterior mediastinal, bronchial, lung (incise every few cm). Abdomen - hepatic portal, mesenteric, ileocaecal, liver, spleen. Carcass- prescapular, superficial inguinal (supramammary or scrotal), internal and medial iliac, deep inguinal, prefemoral, popliteal.
• Collect all tissues listed in bold type and a sample of each lesion found and surrounding tissues, especially those that range from white, grey, or yellow nodules with caseous, necrotic centres, to purulent thin-walled abscesses (particularly in Cervidae and Camelidae), any lymphadenitis, granulomas or actino-like lesions.
• No prescreening of lesions is permitted (no use of impression smears, etc.).
• All tissues and lesions (with surrounding tissues) collected have to be submitted for both histopathology and culture. Do not send a lymph node from one side of the animal for histopathology and the corresponding one from the other for culture.
• Place a representative portion of each tissue and lesion in 10% buffered (pH 7.0) formalin. The proportion of tissues to formalin should not exceed 1 part tissue to 10 parts formalin. Use one jar per animal except if the volume of tissues exceeds the 1/10 ratio. Tape around seam of the lid (tape in the direction of the closure).
• If possible and practical, sort the tissues for culture in 4 groups; head, thorax, abdomen, carcass. Always pack lesions separately. Place the tissue in a high density polyethylene bag, such as a Whirl-Pak. Do not use Zip-lock bags. Remove any air and seal the bag (never staple). If using a Whirl-Pak, fold approximately 4 times before bending in the tabs. Package the tissues from different animals separately, in different bags.
• You can use the contents of one of the new TB Submission Kits but the outer packaging must be as in Section 3.
• Using permanent ink label all bags and jar(s) with the Health of Animals ear tag number, Canadian Cattle Identification Program tag number or other appropriate/traceable animal identification.
• Write a list and a description (location, appearance, size, consistency) of the tissue(s) and lesion(s) on the submission form. Use a separate submission form for each animal but the same form (reference number) for formalin and culture samples from the same animal.

2. Storage of samples after collection

If the tissues will be received at the laboratory within 48 hrs of collection, send them refrigerated with ice-packs. If the period will exceed 48 hrs, freeze the tissue for culture and when ready, ship frozen with ice-packs. Do not freeze the tissues in formalin.

3. Packaging

• The samples must be packaged by CFIA staff holding a valid Transportation of Dangerous Goods (TDG) Training Certificate. All conditions included in the Permit for Equivalent Level of Safety, TC-SU0286, for Diagnostic Specimens Risk Group 3 must be met as well as any other applicable TDG requirements.
• Place tissue bags and absorbent material into a second high density plastic bag, remove excess air and seal.
• Place each formalin jar and an absorbent packet into a high density plastic bag and seal.
• Pack ice packs, bags and jars into a rabies can (permanently marked TC-125-1B). Pad well to prevent shifting.
• If necessary, formalin samples may be shipped separately from culture tissues (there is too much to fit into the rabies can for example). A copy of the culture tissue submission form (with the same reference number) must accompany the formalin shipment. If the proportion of sample to formalin is 1:10 or less, ship as a diagnostic sample (See "Instructions for the use of the Tuberculosis Submission Kit"). If greater than 1:10 package and ship as dangerous goods.
• All documents, including a current copy of Permit TC-SU0286 and the submission form must be placed on the outside of the rabies can.

4. Shipping

All samples must be shipped so as to minimize transport time (overnight courier). Do not ship late in the week to avoid samples being held over the weekend. For urgent cases or more information contact the Diagnostic Coordinator or the Tuberculosis Laboratory at CFIA Ottawa Laboratory (Fallowfield) Tel: 613-228-6698, Fax: 613-228-6668.

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