Prevalence and Distribution of African Swine Fever Virus in Swine Feed After Mixing and Feed Batch Sequencing

Authors

C. Grace Elijah, Kansas State UniversityFollow
Jessie D. Trujillo, Kansas State UniversityFollow
Cassandra K. Jones, Kansas State University, ManhattanFollow
Taeyong Kwon, Kansas State UniversityFollow
Charles R. Stark, Kansas State University, ManhattanFollow
Konner R. Cool, Kansas State UniversityFollow
Chad B. Paulk, Kansas State UniversityFollow
Natasha N. Gaudreault, Kansas State UniversityFollow
Jason C. Woodworth, Kansas State UniversityFollow
Igor Morozov, Kansas State UniversityFollow
Carmina Gallardo, Instituto Nacional de Investigación y Technología Agraria y Alimentaria, Animal Health Research Centre, Madrid, Spain
Jordan T. Gebhardt, Kansas State University, ManhattanFollow
Jurgen A. Richt, Kansas State UniversityFollow

Keywords

swine, African swine fever virus, feed safety

Abstract

As the United States maintains trade with countries where African swine fever virus (ASFV) is endemic, it is critical to have methods that can detect and mitigate the risk of ASFV in potentially contaminated feed or ingredients. Therefore, the objectives of this study were to 1) evaluate feed batch sequencing as a mitigation technique for ASFV contamination in a feed mill, and 2) determine if a feed sampling method could identify ASFV following experimental inoculation. Batches of feed were manufactured in a BSL-3Ag room at Kansas State University’s Biosafety Research Institute in Manhattan, KS. First, the pilot feed manufacturing system mixed, conveyed, and discharged an ASFV-free diet. Next, a diet was manufactured using the same equipment, but contained feed inoculated with ASFV for a final concentration of 5.6 × 10⁴TCID₅₀/g. Then, four subsequent ASFV-free batches of feed were manufactured. After discharging each batch into a biohazard tote, 10 samples were collected in a double ‘X’ pattern. Samples were analyzed using a qPCR assay specific for the ASFV p72 gene to determine the cycle threshold (Ct) and log₁₀genomic copy number (CN)/g of feed. Batch of feed affected the qPCR Ct values (P<0.0001) and the log₁₀genomic CN/g (P<0.0001) content of feed. Feed samples obtained after manufacturing the ASFV-contaminated diet contained the greatest (P<0.05) amounts of ASFV p72 DNA across all criteria. Quantity of ASFV p72 DNA decreased sequentially as additional batches of initially ASFV-free feed were manufactured, but it was still detectable after batch sequence 4, suggesting cross contamination between batches. This subsampling method was able to identify ASFV genetic material in feed samples using the PCR assay specific for the ASFV p72 gene. In summary, sequencing batches of feed decreases concentration of ASFV contamination in feed, but does not eliminate it. Bulk ingredients or feed can be accurately evaluated for ASFV contamination by collecting 10 evenly distributed subsamples, representing 0.05% of the volume of the container, using the sampling method described herein.

Recommended Citation

Elijah, C. Grace; Trujillo, Jessie D.; Jones, Cassandra K.; Kwon, Taeyong; Stark, Charles R.; Cool, Konner R.; Paulk, Chad B.; Gaudreault, Natasha N.; Woodworth, Jason C.; Morozov, Igor; Gallardo, Carmina; Gebhardt, Jordan T.; and Richt, Jurgen A. (2021) "Prevalence and Distribution of African Swine Fever Virus in Swine Feed After Mixing and Feed Batch Sequencing," Kansas Agricultural Experiment Station Research Reports: Vol. 7: Iss. 10. https://doi.org/10.4148/2378-5977.8150