Sampling and detection of African swine fever virus within a feed manufacturing and swine production system.

Transmission of biological hazards capable of causing disease in livestock can occur through a wide variety of direct and indirect routes. In swine production, there are a large number of possible routes of exposure of a pathogen into a susceptible population. African swine fever virus (ASFV) has been a significant challenge for Southeast Asia since first detected in China in 2018 and has spread through many countries within the region. In order to understand potential transmission pathways within an ASFV endemic region, a diagnostic investigation was performed to determine the level of contamination on a wide variety of surface types within a live animal production, feed manufacturing, and feed distribution system located in Vietnam. All diagnostic testing was performed locally by the production system's internal diagnostic laboratory using real-time polymerase chain reaction (rt-PCR) analysis. Early in the diagnostic investigation, it became clear that feed trucks were a common site of ASFV surface contamination detection. This information resulted in biosecurity-focused actions for feed trucks arriving back at the feed mill, including decontamination of interior truck cab surfaces and washing of exterior truck surfaces with high-pressure water prior to application of surface disinfectants. Additionally, a low number of rt-PCR positive samples were detected within the feed production system, with the greatest number coming from transient surfaces such as high traffic areas and worker clothing. This illustrates the importance of managing employee traffic through procedures such as zoning and separation between clean-dirty areas to reduce the likelihood of pathogen transmission. In conclusion, this report illustrates the importance of routine data capture regarding efficacy of biosecurity procedures which allows for real-time updates and improvement as biosecurity gaps are identified.

Evaluating the distribution of African swine fever virus within a feed mill environment following manufacture of inoculated feed.

It is critical to understand the role feed manufacturing may have regarding potential African swine fever virus (ASFV) transmission, especially given the evidence that feed and/or ingredients may be potential vectors. The objective of the study was to evaluate the distribution of ASFV in a feed mill following manufacture of contaminated feed. To accomplish this, a pilot-scale feed mill consisting of a mixer, bucket elevator, and spouting was constructed in a BSL-3Ag facility. First, a batch of ASFV-free feed was manufactured, followed by a batch of feed that had an ASFV-contaminated ingredient added to feed, which was then mixed and discharged from the equipment. Subsequently, four additional ASFV-free batches of feed were manufactured using the same equipment. Environmental swabs from 18 locations within the BSL-3Ag room were collected after each batch of feed was discharged. The locations of the swabs were categorized into four zones: 1) feed contact surface, 2) non-feed contact surface < 1 meter away from feed, 3) non-feed contact surface > 1 meter from feed, and 4) transient surfaces. Environmental swabs were analyzed using a qPCR specific for the ASFV p72 gene and reported as genomic copy number (CN)/mL of environmental swab processing buffer. Genomic copies were transformed with a log10 function for statistical analysis. There was no evidence of a zone × batch interaction for log10 genomic CN/mL (P = 0.625) or cycle threshold (Ct) value (P = 0.608). Sampling zone impacted the log10 p72 genomic CN/mL (P < 0.0001) and Ct values (P < 0.0001), with a greater amount of viral genome detected on transient surfaces compared to other surfaces (P < 0.05). This study illustrates that once ASFV enters the feed mill environment it becomes widespread and movement of people can significantly contribute to the spread of ASFV in a feed mill environment.