Location: Exotic & Emerging Avian Viral Diseases Research
Project Number: 6040-32000-081-037-I
Project Type: Interagency Reimbursable Agreement
Start Date: May 15, 2024
End Date: Sep 30, 2024
Objective:
To characterize highly pathogenic avian influenza in dairy cattle, including detection of virus, sequence analysis, and survival of virus in milk. This will include determining the thermal inactivation profile of highly pathogenic avian influenza virus in dairy products using different pasteurization methods, including continuous flow pasteurization. Evaluate commercially available lateral flow assays (LFA) to determine sensitivity, specificity and ease of use as a point of care tests of milk at the cow level and the bulk milk tank level. Development of methods to optimize detection and sequencing of HPAI from milk, and the analysis of the information gathered.
Approach:
Thermal inactivation of highly pathogenic avian influenza in whole milk will be evaluated by different methods. Laboratory methods will be evaluated on small milk samples using thermal heat blocks at different times and temperatures will be evaluated. More robust characterization will be conducted using a laboratory scale continuous flow pasteurizer that closely mimics commercial equipment will be used to evaluate the most common times and temperatures for the pasteurization in milk. Effectiveness of pasteurization will be evaluated by growing virus in milk in a quantitative manner in embryonating chicken eggs. Both retail milk and milk products and bulk milk samples will also be evaluated for viral RNA detection and the results compared with virus detection in embryonating chicken eggs. These samples will be collected by Food and Drug Administration and provided to ARS. Commercially available lateral flow assays (LFA) will be evaluated for sensitivity, specificity, and ease of use. These samples will be compared with spiked samples of different viruses with known virus quantities to allow comparisons of tests. A smaller subset of tests will be evaluated with fresh milk samples from cows. Finally, updated test procedures to extract RNA from milk and milk samples will be conducted to determine best methods. Additional sequencing and sequence analysis will be conducted to determine best methods to work with milk and milk samples.
