Analysis of acidified feed components containing African swine fever virus

Mitigation of African swine fever (ASF) virus in contaminated feed materials would assist control activities. Various finely-ground pig feed ingredients (5 cereals, 4 plant proteins, 2 animal proteins, 1 oil, 1 compound) were sprayed and mixed thoroughly with a buffered formic acid formulation (0, 1 or 2% vol/vol) to produce a consistent and durable level of formate (1% or 2%) with consistent acidification of cereal ingredients to less than pH 4. No such acidification was noted in other ingredients. Selected representative feed ingredients were further mixed with infectious ASF virus (106 TCID50) or media alone and incubated for 0, 6, 12, 24, 48, 72 or 168 h. The residual ASF virus at each timepoint was quantified using qPCR and a cell culture based TCID50 assay to determine survivability. Maize, rice bran and compound feed (with or without formate) all reduced infectious ASF virus to levels below the detection threshold of the cell culture assay (101.3 TCID50/mL). A consistent reduction in ASF virus DNA levels was observed by qPCR assay when maize containing ASF virus was mixed with 1% or 2% buffered formic acid. This reduction in viral DNA corresponded to the acidifying pH effect measured. No such reduction in ASF virus DNA levels was noted in non-cereal ingredients containing ASF virus, in which the pH had not been lowered below pH 4 following treatment. Interestingly, residual ASF virus levels in spiked meat/bone meal were greater than control levels, suggesting a buffering effect of that feed ingredient.