Chagas disease is considered a public health issue in Colombia, where many regions are endemic. Triatoma dimidiata is an important vector after Rhodnius prolixus , and it is gaining importance in Boyacá, eastern Colombia. Following the recent elimination of R . prolixus in the region, it is pivotal to understand the behavior of T . dimidiata and the transmission dynamics of T . cruzi . We used qPCR and Next Generation Sequencing (NGS) to evaluate T . cruzi infection, parasite load, feeding profiles, and T . cruzi genotyping for T . dimidiata specimens collected in nine municipalities in Boyacá and explored T . dimidiata population genetics. We found that T . dimidiata populations are composed by a single population with similar genetic characteristics that present infection rates up to 70%, high parasite loads up to 1.46 × 10 9 parasite-equivalents/mL, a feeding behavior that comprises at least 17 domestic, synanthropic and sylvatic species, and a wide diversity of TcI genotypes even within a single specimen. These results imply that T . dimidiata behavior is similar to other successful vectors, having a wide variety of blood sources and contributing to the circulation of different genotypes of the parasite, highlighting its importance for T . cruzi transmission and risk for humans. In the light of the elimination of R . prolixus in Boyacá and the results we found, we suggest that T . dimidiata should become a new target for vector control programs. We hope this study provides enough information to enhance surveillance programs and a future effective interruption of T . cruzi vector transmission in endemic regions.