Characteristics of Trypanosoma cruzi infection were studied in a rural area of the eastern plains of Colombia. Using enzyme-linked immunosorbent assay and indirect fluorescent-antibody tests, the infection was determined in 11.6% of the inhabitants of 142 dwellings. During 6 months of community surveillance, in 42.3% dwellings, 609 triatomines were collected (597 Rhodnius prolixus and seven, three, one, and one of Panstrongylus geniculatus, Psammolestes arturi, Eratyrus mucronatus, and Triatoma maculata, respectively). Rhodnius prolixus was found in 80% peridomiciliary Attalea butyracea palms examined with baited traps, and its infection with T. cruzi was 30% and 38.5% in dwellings and palms, respectively. Trypanosoma cruzi was isolated in five of 35 triatomines and in one of 24 dogs. The blood of domestic and wild animals was identified in triatomines collected in the intradomicile and in palms. These results support the extension of the wild cycle of T. cruzi to human dwellings and the characterization of a new scenario for transmission in Colombia.