In Colombia, dogs and opossum are the most important mammals in domestic and sylvatic T. cruzi transmission. However, the role of both species has not been evaluated in areas where both species converge in the peridomestic area. To evaluate the infection status of domestic and wild mammals in peridomestic habitats of Puerto Valdivia, Antioquia Department. The infection of domestic dogs and small wild mammals was performed by hemoculture, molecular and serological methods. Additionally, the infection in children under 15 years old and triatomine searches was carried out. We found that 16.07% and 34% dogs, and 59.1% and 61.1% Didelphis marsupialis were found positive by molecular and serological methods respectively. Moreover, in 25% and 75% of the infected dogs were detected TcI Dom and TcI sylvatic, respectively, while all the D. marsupialis were infected with TcI. Six Rattus rattus and three Proechimys semispinosus were captured but without T. cruzi infection. Finally, none of the 82 children were positive and no triatomine bugs were captured. D. marsupialis and domestics dogs have an important role in the transmission of T. cruzi suggesting a potential risk in T. cruzi transitions areas.