Gut microbiota, blood feeding sources, and Trypanosoma cruzi infection in Rhodnius colombiensis from center-west of Colombia

Chagas disease, caused by Trypanosoma cruzi, remains a major public health concern in Latin America. In Tolima, Colombia, the vector R. colombiensis has acquired epidemiological relevance due to its ability to colonize domiciliary environments and its high rates of T. cruzi infection, which increase transmission in areas where primary vectors such as R. prolixus have been controlled. In this study, we characterized the gut microbiome of R. colombiensis using culture-dependent and culture-independent approaches, including next-generation sequencing (NGS) of the 16S rRNA gene, specific detection of bacterial endosymbionts, identification of blood meal source, and molecular screening for T. cruzi. DNA Analysis of 151 specimens revealed Didelphis marsupialis and Gallus gallus as primary blood meal sources. DNA of T. cruzi was detected in 92% of samples, along with endosymbionts such as Cardinium spp. and the intracellular parasite Microsporidia (2.2%-4,4%). The phyla Actinobacteria, Firmicutes, and Proteobacteria represented the dominant microbial community, with Gordonia spp., Lactococcus spp., and Enterobacter spp. as the predominant genera. Culture-based analyses revealed additional prevalent genera, including Staphylococcus sp. and Yokenella sp., which may play a role in microbial competition within the triatomine gut. Our findings provide the first comprehensive characterization of the gut microbiota, endosymbionts, blood meal sources, and T. cruzi infection in R. colombiensis, contributing to a better understanding of the vector-microbiota-parasite interactions relevant to the transmission of Chagas disease.