Abstract
The Triatoma rubrovaria subcomplex, comprising several triatomine species, plays a significant role in the transmission of Chagas disease in southern Brazil. Despite morphological distinctions among these species, their genetic differentiation remains poorly understood, particularly in sympatric regions. This study investigates the genetic diversity and phylogenetic relationships through DNA sequencing analysis of five sympatric species within the T. rubrovaria subcomplex (T. rubrovaria, T. carcavalloi, T. klugi, T. circummaculata, and T. pintodiasi), using a 542-bp fragment of the mitochondrial cytochrome b (mtCytb) gene. A total of 84 specimens were collected from six municipalities in Rio Grande do Sul, Brazil, and analyzed alongside laboratory-reared specimens and sequences from the GenBank. Bayesian phylogenetic reconstructions, haplotype networks, and population structure analyses revealed a lack of clear genetic differentiation among the five species, with overlapping intra- and interspecific divergences and shared haplotypes. These findings suggest either a single species exhibiting phenotypic plasticity or a group of incipient species with ongoing gene flow. This study highlights the need for a taxonomic revision and suggests that this group could serve as a valuable model for further genomic research to elucidate potential aspects of phenotypic plasticity and/or sympatric speciation in triatomines.