Entomological surveys were conducted in five rural communities (138 domiciliary units [DUs]) in the southern Andes of Ecuador. Adobe walls and ceramic tile roofs were predominant construction materials. A 35% house infestation rate with Panstrongylus chinai (Del Ponte, 1929) (0.7%), Panstrongylus rufotuberculatus (Champion, 1899) (0.7%), Rhodnius ecuadoriensis (Lent & León, 1958) (27%), and/or Triatoma carrioni (Larrousse, 1926) (7%) was found. Adults and nymphs of R. ecuadoriensis and T. carrioni were found in intradomiciliary and peridomiciliary areas. Breeding triatomine colonies were present in 85% of infested DUs, and the average insect crowding was 52 ± 113 triatomine bugs per infested house. T. cruzi-like organisms were found by microscopic examination in the feces or hindgut but not the salivary glands of 4% of examined R. ecuadoriensis and 12% T. carrioni. Serological tests detected a general anti-T. cruzi antibody seroprevalence of 3.9% (n = 1136). Only 2% of individuals had heard of Chagas disease, and although triatomines were reported as a major nuisance by the population they were not considered vectors of disease. Additional baseline field research is needed for the design and implementation of a Chagas disease control program in the region.