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This work was supported by The Pro-Amaz?nia Biodiversity and Sustainability project was financed by the Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior ? Brasil (CAPES), notice 047/2012, AUXPE 3286/2013 - Process 23,038.009430 / 2013?98. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Laborat?rio de Geoprocessamento. Ananindeua, PA, Brasil. Universidade Federal do Par?. Instituto de Ci?ncias Biol?gicas. Laborat?rio de Ecologia e Zoologia de Vertebrados. Bel?m PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Castanhal, PA, Brazil. Chagas disease is an anthropozoonosis, caused by a flagellated protozoan, Trypanosoma cruzi, in which the enzootic cycle occurs between mammals and triatomines. Two dogs with a history of sudden death were necropsied at the Federal University of Par? (UFPA). One dog had a pale area in the myocardium, which on histopathological examination showed a T. cruzi amastigote nest; immunohistochemistry (IHC) analysis characterized it as acute Chagas disease (ACD). The second dog showed no macroscopic changes. Microscopically, a few cardiomyocytes were replaced by adipocytes, and IHC result was negative for T. cruzi. However, results of polymerase chain reaction (PCR) of the cardiac tissue of both dogs was positive for T. cruzi DNA. After that, an epidemiological study was conducted in the region. For this study, we selected four areas in Castanhal. One of the four areas (Area 1) is where one of the dogs lived. The other three areas were chosen because they were recently deforested for housing. Blood samples were collected from dogs, cats, wild small mammals (marsupials and rodents), and the digestive tract of triatomines. Nested PCR was performed on all the blood samples and the triatomine digestive tracts. In Area 1, T. cruzi DNA was detected in 50% (12/24) of the tested dogs, in the only tested cat (1/1), 50% (1/2) of the tested marsupials (Didelphis marsupials), and 100% of the captured triatomines (Rhodnius pictipes) (2/2). In Area 2, T. cruzi DNA was not detected in any of the 11 (0/11) dogs and two marsupials tested (0/2), and no triatomines were found in this area. In Area 3, T. cruzi DNA was detected in 42.25% (30/71) of the dogs, in 66,6% (2/3) of the cats, the only captured marsupial (D. marsupialis) (1/1), and all three triatomines (3/3) (R. pictipes) tested. In Area 4, the two dogs tested were negative (0/2), 25% (1/4) of the captured marsupials (D. marsupialis) was positive, and no triatomine was captured in this area. The data demonstrate the importance of detecting T. cruzi in dogs, cats, small rodents, and marsupials in the Amazon metropolitan areas, where ecotopes carry reservoirs and vectors capable of participating in the Chagas disease cycle. The proximity between humans and T. cruzi vectors in these places might contribute to increased disease transmission risk and maintenance of agents. It was noted that high-standard condominiums, previously thought to reduce the risk for this disease, presented a new epidemiological risk. The presence of T. cruzi DNA in a dog who, a year earlier had tested negative, when another dog in the same house died of ACD, shows that the transmission cycle is present and active, with a high possibility of disease transmission to animals and humans.