Viral metagenome characterization reveals species-specific virome profiles in Triatominae populations from the southern United States

Kissing bugs (Triatominae) are hematophagous insects and the principal vectors of Trypanosoma cruzi , the causative agent of Chagas disease. While their bacterial microbiomes have received considerable attention, the diversity of viruses associated with these insects remains poorly understood. To address this gap, we investigated the metavirome of five Triatominae species from the southern United States ( Triatoma rubida, T. sanguisuga, T. gerstaeckeri, T. indictiva , and Hospesneotomae protracta ), sampled in Texas, New Mexico, and Arizona. We sequenced gut and reproductive tissues from 23 field-collected specimens and assembled 41 viral operational taxonomic units (vOTUs), 40 of which are novel and together constitute 13 viral families, including Chuviridae, Arenaviridae, Orthomyxoviridae, Partitiviridae, Solemoviridae, Circoviridae, Rhabdoviridae, Microviridae, Xinmoviridae, Astroviridae, Narnaviridae, Tombusviridae , and the order Elliovirales. The vOTUs composition and abundance analysis examined variables including host species, sex, tissue type, blood meal, and T. cruzi infection status, showing that metavirome diversity varied significantly among host species. Our findings demonstrate a species-specific metavirome and the presence of virus taxa linked to insects, plants, and vertebrates, highlighting the complex ecological interactions between viruses and triatomines. This study uncovers a diverse and largely novel set of metaviromes within North American Triatominae, providing a foundation for future research on virus-vector interactions.