Viral metagenomics reveals parvovirus dark matter of herbivorous wildlife from the Qinghai-Tibet Plateau

As one of the birthplaces of modern biodiversity, the Qinghai-Tibet Plateau is home to a rich variety of unique animal groups. Herbivorous wildlife constitutes a vital component of the plateau's ecosystems, yet current research on parvoviruses in these species remains limited. With the development of viral metagenomics and next-generation sequencing technology, more and more novel viruses have been detected. In this study, we collected 741 fecal samples from herbivorous wildlife across three distinct habitats on the Qinghai-Tibet Plateau and employed metagenomics analysis technology to characterize the parvovirus dark matter. We identified 32 parvoviruses by analysis, of which 13 were closely related to the members of Bocapavovirus, Dependoparvovirus and Protoparvovirus in the subfamily of Parvovirinae, 5 belonging to the Densovirinae subfamily, and 5 were classified into the newly proposed subfamily of Hamaparvovirinae. Additionally, 9 parvoviruses remain unclassified and cannot be assigned to any existing subfamily. We conducted a phylogenetic analysis based on the non-structural proteins of the viruses to explore the evolutionary relationships among parvoviruses, and found that some viruses could not be clustered with known parvoviruses. These results enrich our understanding of parvovirus and viral dark matter, and reveal that these wild animals carry a large number of unknown new viruses that may have implications for the ecological balance of the Qinghai-Tibet Plateau.