One of the most daunting challenges for Chagas disease surveillance and control in Mexico is the lack of community level data on vector distributions. Although many states now have assembled representative domestic triatomine collections, only two triatomine specimens had been collected and reported previously from the state of Guanajuato. Field personnel from the state’s Secretaría de Salud conducted health promotion activities in 43 of the 46 counties in the state and received donations of a total of 2,522 triatomine specimens between 1998 and 2002. All specimens were identified, and live insects examined for Trypanosoma cruzi. In an effort to develop fine-scale distributional data for Guanajuato, collection localities were georeferenced and ecological niches were modeled for each species by using evolutionary-computing approaches. Five species were collected: Triatoma mexicana (Herrich-Schaeffer), Triatoma longipennis (Usinger), Triatoma pallidipennis (Stål), Triatoma barberi (Usinger), and Triatoma dimidiata (Latreille) from 201 communities located at elevations of 870–2,200 m. Based on collection success, T. mexicana had the broadest dispersion, although niche mapping indicates that T. barberi represents the greatest risk for transmission of Chagas disease in the state. T. dimidiata was represented in collections by a single adult collected from one village outside the predicted area for all species. For humans, an estimated 3,755,380 individuals are at risk for vector transmission in the state, with an incidence of 3,500 new cases per year; overall seroprevalences of 2.6% indicate that 97,640 individuals are infected with T. cruzi at present, including 29,300 chronic cases.