Detection of African swine fever virus and wild boar eDNA in soil and turbid water samples: towards environmental surveillance

Since 2007, an ongoing African swine fever (ASF) pandemic has significantly impacted Eurasia. Extensive field evidence and modeling confirm the central role of wild boar in ASF epidemiology. To effectively control and eradicate the infection, rapid detection of the ASF virus (ASFV) is crucial for prompt intervention in areas of recent viral introduction or ongoing outbreaks. Environmental DNA (eDNA) is a cost-effective and non-invasive technique that has shown promising results in monitoring animal species and their pathogens and has the potential to be used for wildlife disease surveillance. In this study, we designed and evaluated an eDNA sampling method for highly turbid water and soil samples to detect ASFV and wild boar (Sus scrofa) DNA as a control using qPCR while ensuring biosafety measures and evaluating ASF epidemiology. To validate our method, we obtained samples from La Mandria Regional Park (LMRP) in northwestern Italy, an area free of ASFV, and spiked them in a laboratory setting with an ASFV’s synthetic DNA template. Our findings highlight the potential of eDNA monitoring as a reliable, rapid, and safe method for early detection of ASFV from soil and turbid water samples.