Ticks within the species complex Rhipicephalus sanguineus sensu lato are carriers for various pathogens worldwide. Among them are some RNA viruses, who have gained increased attention due to their potential for interaction between pathogenic and non-pathogenic organism. Our study aimed to investigate the RNA virus composition in R. sanguineus s.l. (also known as R. linnaei) ticks collected from the Santander and Casanare regions in eastern Colombia. We employed Oxford Nanopore sequencing technology coupled with viral enrichment procedures. Thirty-nine adult ticks, removed from dogs, were sequenced with Oxford Nanopore sequencing. These reads underwent two distinct analyses: one focused on sequencing reads utilizing Centrifuge/BLAST for direct assignment, and one involved assembly of viral metagenomic assembled genomes (vMAGs) using the Genome Detective Tool program. Our investigation showed the presence of six distinct viruses within the tick virome: Trinbago virus, Mivirus sp., Bole tick virus 4, Brown dog tick phlebovirus 2, Totiviridae sp., Ixodes scapularis totilike virus, and Brown dog tick phlebovirus 1. Notably, we successfully assembled the genomes for the latter two species, enabling phylogenetic and comparative analyses with genomes from various global regions. Only the category of sex yield significant differences in alpha diversity. We described viruses with potential pathogenicity but also with endosymbiotic potential. The latter group holds promise for developing biotechnological tools to aid future tick vector control strategies. Furthermore, our research offered valuable insights into the predominant viruses found in R. sanguineus s.l. ticks in two previously unexplored regions within Colombia.
