Supplies needed:

1. AI Surveillance kit (swabs, BHI media tubes, mailer with postage paid mailing label). Refrigerate BHI media before and after use.
2. AI Lab Submission Form.
3. Cooler with ice packs.
4. Exam gloves, hand soap, disinfectant, water.
5. Sharpie or bar code labels.
6. Trash bag.

Biosecurity considerations:

While sampling for AI should cause no risk to human health, care should be taken by the collector to prevent the spread of infectious disease. Always wear clean coveralls, rubber boots, and gloves at each testing site. Take care not to touch your eyes or mouth until removing gloves and washing hands. These precautions will limit exposure to other infectious organisms as well. After collecting samples, remove coveralls and disinfect equipment (including boots) prior to approaching the vehicle. This will prevent contamination of the vehicle and other equipment. Dispose of gloves, swabs, and other trash at a veterinary clinic or home waste receptacle on the premises. Wash your hands immediately after removing gloves from sampling.

If collecting samples on farm at multiple premises, use a separate AI surveillance kit for each premise. Prior to entering the facilities to collect samples, ask the owner how many birds of each species are on the premises to determine the number of samples to be collected, up to a maximum of 30 samples per site. Unused tubes and swabs should be considered contaminated and disposed of properly or returned in the AI Surveillance Kit. Collect samples from all avian species, except pigeons and doves, on the premises.

For exhibits (including county fairs), swap meets, and live bird market sampling, each exhibitor's or consignor's birds should be sampled even if they are from the same family or premises. Contact the event organizer or superintendent to determine the best time to sample poultry. Collect samples from all avian species, except pigeons and doves. Do not collect samples from waterfowl at county fairs. Every attempt should be made to collect five (5) samples from each consigner/exhibitor, up to 30 birds maximum. It is acceptable to group five (5) samples per species and submit them in one tube. Indicate species and consignor/exhibitor number next to the bar code label or tube number on Lab Submission Form

Tracheal Sampling Procedures:

To collect tracheal swab samples, hold the bird's head up in a nearly vertical position with wings and feet contained. The bird should face the person swabbing the trachea. Remove swab from package and insert tip into tracheal orifice (1cm). Rotate swab tip against tracheal lining two or three times, and place directly into liquid transport media (BHI broth). With the swab in the media, swirl the stem end of the swab between fingers vigorously, and leave the swab in the tube. Repeat this procedure for up to five (5) swabs from the same exhibitor/owner per tube. When all swabs are in the media, slowly turn and pull all swabs out of the tube at the same time, causing the contents to be expressed into the tube. Dispose of swab properly (do not leave in tube).

Cloacal Sampling Procedures:

To collect cloacal swab samples, hold the bird's head down in a nearly vertical position with wings and feet contained. The bird's ventrum should face the person swabbing. Locate and grasp tail feathers at the base and reflect away from you to locate cloacal. Remove swab from package and insert tip into cloacal orifice (1cm). Rotate swab tip against cloacal lining two or three times. Remove swab, shake off excess fecal material, and place directly into liquid transport media (BHI broth). With the swab in the media, swirl the stem end of the swab between fingers vigorously, and leave the swab in the tube. Repeat this procedure for up to five (5) swabs from the same exhibitor/owner per tube. When all swabs are in the media, slowly turn and pull all swabs out of the tube at the same time, causing the contents to be expressed into the tube. Dispose of swab properly (do not leave in tube).

Blood Sampling Procedures:

Using a small syringe (with a 20 or 21 gauge needle), collect a 1.5 to 2.0 ml sample of blood from the large median wing vein. Inject the blood into a red-top, serum separater tube. Following collection, place the tubes in a slanted position or on their sides until the clot forms. After the clot forms, place the tubes in a centrifuge and spin for 10 minutes at 3,000 rpms. Take the tubes from the centrifuge and place in a brucellosis box for shipment to the lab.

Fecal Sampling Procedures:

Swabs of fecal material may be taken at exotic sales or auctions where access to individual birds is limited. Identify all birds/cages belonging to one consignor. Rotate the sterile swab in feces dropped below the bird or on the edge of the cage. Remove swab, shake off excess fecal material, and place directly into liquid transport media (BHI broth). With the swab in the media, swirl the stem end of the swab between fingers vigorously, and leave the swab in the tube. Repeat this procedure for up to five (5) swabs from the same exhibitor/owner per tube. When all swabs are in the media, slowly turn and pull all swabs out of the tube at the same time, causing the contents to be expressed into the tube. Dispose of swab properly (do not leave in tube).

Submission paperwork and shipping procedures:

1. Complete Laboratory Submission Form noting tube number (or bar codes where available), species, consignor/exhibitor name or number (for county fair and exotic sales), and number of birds sampled in each tube. Private practitioners should remember to check the Fee Basis Box (See Guidelines for Fee Basis Veterinarians - AI11).

2. Place tubes in brucellosis tube box (slotted cardboard box).

3. Place the completed and signed white copy of the AI Laboratory Submission Form in the brucellosis tube box.

4. Place box into plastic bag with an absorbent sheet and seal.

5. Place two ice packs inside shipping container and place the sealed plastic bag between the ice packs.

6. Close and seal the outer cardboard shipping container.

7. Attach shipping label to top of box.

8. Attach Diagnostic Specimen label (UN3373) to side of box.

9. Samples should reach UNL-VDC within four days of collection if possible. Do NOT mail samples on Friday or Saturday.

10. Forward the yellow copy of Laboratory Submission Form to BAI Office and retain the pink copy for your records.

11. UNL-VDC will mail results to BAI Office.

Fee basis payments will be processed once the Laboratory Submission Form and test results are received in the BAI office. For questions about sampling poultry for AI, please contact the BAI office at (402) 471-6880.

(Rev 04/08)