Non-Invasive Surveillance of the Wildlife Virome in Central Italy: Insights from the Foreste Casentinesi National Park

Wildlife can act as both a reservoir and a sentinel for emerging pathogens, but surveil-lance is often constrained by difficulties in obtaining samples without disturbing animals. This study explored the viral diversity of wild mammals inhabiting the Foreste Casen-tinesi National Park (Central Italy) using non-invasive fecal sampling. From 2021 to 2022, 99 fecal samples from several species were collected and analyzed by PCR and meta-genomic next-generation sequencing. Of 26 pools examined, 10 (38.5%) tested positive for at least one viral target. Astroviruses were the most frequently detected, found in deer, foxes, wolves, small mustelids, and porcupines. Foxes carried the widest range of viruses, including astrovirus, parvovirus, bocavirus, kobuvirus, adenovirus, and coronavirus. Several sequences showed low similarity to known strains, suggesting divergent or novel viral lineages. Metagenomic analysis also identified members of Circoviridae, Anelloviridae, and Picobirnaviridae. These results provide new insights into the virome of European wild-life, including the first reports of some viruses in certain species. Overall, our study demonstrates that non-invasive surveillance is a valuable tool for monitoring ecosystem health and supports a One Health approach to early detection of viral threats.