Sample Collection, Testing, and Surveillance for Highly Pathogenic Avian Influenza (HPAI)

Sampling for HPAI

The rRT-PCR test is commonly used to diagnose and monitor High Pathogenic Avian Influenza (HPAI). Sample collection must be performed by Category II Accredited Veterinarians, Agents of the State, or Maryland Authorized Poultry Testing Agents. When sampling gallinaceous poultry (chickens, turkeys, pheasants, quail), an oropharyngeal or tracheal swab should be taken. For domestic waterfowl (ducks and geese), a cloacal swab should be collected.

Polyester, rayon, or other inert swabs with plastic handles are recommended for sample collection. To order or pick up BHI from Salisbury Animal Health Laboratory or Frederick Animal Health Laboratory, call with request 7-10 days prior to need. BHI can be stored in a home refrigerator or freezer. It's advised to prioritize sampling fresh mortalities, followed by sick birds and then healthy birds, to achieve the minimum sample size.

Maryland Department of Agriculture Guidelines for Proper Sample Collection

• Unused BHI broth media taken into the field and not used must be disposed.
• Do not use the BHI if it appears cloudy or is less in volume than the recommended 3 ml.
• Only one species through one route should be pooled in a sample collection tube.
• No less than 2 ml of BHI should be remaining in each tube after pooling the swabs. Do not Top-off.
• The tube should be tightly capped and should be cleaned with Virkon-S or 10% bleach from the outside.
• The tubes should not be stored in a household freezer after swabbing.
• A separate submission form must be submitted for each species of bird.
• Sample arriving without fulfilling the requirements may be rejected and re-submission may be required.
• Samples should preferably be transported overnight in properly chilled condition.

Surveillance of HPAI in HPAI Control Area

Active Surveillance is conducted around the Control Area of HPAI outbreak to confirm the presence or absence of the Virus. This helps uninfected poultry premises to continue their farm operations and obtain movement permits for poultry and poultry products and other necessities. Active Surveillance of the commercial poultry premises has two components: Routine Active Surveillance and Pre-movement Surveillance.

Routine Active Surveillance

Pre-Movement Surveillance

To obtain a permit to move live broilers outside of the Control Area, the producers must produce negative rRT-PCR test results from at least two 11-bird pooled samples from flock. If there are fewer than 22 dead birds, all the dead and sick birds should be swabbed, and the swabs should be collected into two separate pools. The two sample pools can be collected in two different ways to obtain a permit for Broilers movement. Both pools can be collected 24 hours prior to the movement. In an alternative approach, the first pool can be collected 48 hours before the movement and the second pool can be collected 24 hours before movement. 

Laboratory Submission:

MDA Animal Health Diagnostic Labs has two laboratories offered for animal disease investigation - in Salisbury and Frederick. It is recommended that the laboratories be contacted before submitting the samples. The sample should be accompanied by detailed information about the sample in a HPAI sample submission form. The required information includes:

• Owner's name, complete address, and phone number
•  Premise ID and address
• Sample Collector’s name, address, and phone number.
• Species, breed, and age of the birds 
• Sample type: (Cloacal, Tracheal, or Oropharyngeal)

Download the required HPAI laboratory submission form 

Please contact the laboratories before submitting the samples.

Acknowledgment:

This work is supported by the Smith-Lever Special Needs Competitive Grant, project grant number 2022-41210-38062, from the USDA National Institute of Food and Agriculture and the University of Maryland, College Park. ( you can add logos for UMD and USDA NIFA as you see fit)

Project Title: Biosecurity Compliance Audits to Prevent Outbreaks of HPAI and Risk-Based Planning to Improve Outbreak Response.

Project Team: Dr. Nathaniel Tablante,  Dr. Mostafa Ghanem,Dr. Jonathan Moyle, Jennifer Rhodes, and Maegan Purdue, Madhusudan Timilsina