Guidance for Diagnostic Testing for Highly Pathogenic Avian Influenza H5N1 Virus in Cattle

Guidance for Diagnostic Testing for Highly Pathogenic Avian Influenza (HPAI) H5N1 Virus in Cattle

By Nebraska Veterinary Diagnostic Center

Recent USDA-APHIS regulations require that milk samples from lactating dairy cattle be submitted to a laboratory approved by the National Animal Health Laboratory Network (NAHLN) for Influenza A virus (IAV) PCR testing prior to interstate movement. The NVDC is approved by NAHLN to run this testing. If you have clinically ill or sick animals that may require testing, please contact the Nebraska Department of Agriculture (NDA) or USDA for instructions.

Samples are required to have a submission form, either a USDA VS 10-4 or the NVDC Ruminant Diagnostic Form.

  • Sample submissions are required to have premise ID number (PIN) for the location of the animals. Please contact the Nebraska Department of Agriculture if you do not have a PIN.
  • In the history or open text fields of the form indicate that you would like Influenza A testing for movement purposes of lactating dairy cows.
Sample collection

Per USDA and state requirements, samples are to be collected by an accredited veterinarian, someone working under the guidance of an accredited veterinarian, or a sample collector approved by the appropriate state animal health official.

Sample collection and testing must take place no more than seven (7) days prior to interstate movement. Samples to be collected are milk/udder secretion sampled from each quarter, collected in a leakproof tube containing 3-10 mL of total milk sample per cow. Attempts should be made to include approximately equal volumes of milk from each quarter in the tube to make up the 3-10 mL total volume. Recommended collection tubes are a 50 mL conical collection tube  or a 4.5-5 mL collection tube  which can be ordered from NVDC using the Supply Order Form, if needed.

Do not pool samples. Submit samples from individual animals only. Samples should be chilled and shipped overnight with frozen gel packs. Use an insulated shipping container with submission information in a leakproof container (Ziploc/whirlpack). Insulated shipping containers can be purchased from the NVDC using the Supply Order Form.

Sample submission

Samples can be shipped using UPS EZ-ship with a shipping label you can print immediately, or by mailing to:

Nebraska Veterinary Diagnostic Center
University of Nebraska-Lincoln
Room 230A NVDC
4040 East Campus Loop North
Lincoln, NE 68583-0907

Testing times

Results are often available the afternoon of the business day following receipt of the samples, but they could take up to three business days after receipt of the samples.

Testing fees

Fees for HPAI testing of milk from lactating dairy cows for the purpose of interstate movement are paid by the USDA. Collection, shipping, and/or transport costs are not covered by the USDA.

Number to test

For groups/lots of 30 or fewer animals moving interstate, all animals being moved must be tested. If more than 30 animals are moving interstate, then only 30 animals total must be tested.

Additional information

Please refer to the guidance document on APHIS Requirements and Recommendations for Highly Pathogenic Avian Influenza (HPAI) H5N1 Virus in Livestock for State Animal Health Officials, Accredited Veterinarians and Producers

Additional contact information

Nebraska Department of Agriculture (NDA): 402-471-2351