Ecological Factors and Host Community Characteristics as Potential Drivers of Bat RNA Virus Spillover

Bats are reservoirs for many emerging viruses, yet broad-scale spatial patterns of bat viromes and their ecological determinants remain unclear, limiting spillover risk assessment. To address this, we conducted a standardized survey across central to southern China. During 2022-2023, fecal samples were collected from 527 bats representing 17 species at 21 caves in seven provinces. Using each cave as the analytical unit, samples from all species at a site were pooled to construct 21 fecal RNA virome libraries for metatranscriptomic sequencing; viral-like contigs were annotated, and viral alpha/beta diversity was quantified at the genus level. Sites were grouped by geographic distance and latitude to evaluate spatial differentiation and to relate virome patterns to host community attributes and environmental factors. We annotated 56 viral families, including 19 vertebrate-associated families, with multiple families and genera shared across geographic groupings. Vertebrate-associated viral diversity showed limited evidence of geographic isolation within the study region but suggested gradual turnover with latitude. Host species richness was the strongest correlate of virome diversity, exceeding the effects of evaluated environmental variables (e.g., climate and human activity). These results support prioritization of species-rich bat habitats for surveillance and risk assessment.