<p><em>Trypanosoma cruzi</em> the causative agent of Chagas disease, can be found in more than 150 different species of domestic and wild mammals in the American continent. Domestic mammals like dogs, and commensal rodents, have been frequently implicated as hosts and/or reservoirs in the domestic and peridomestic cycle of transmission, however, little attention have been paid to backyard mammals as potential maintaining hosts of <em>T. cruzi</em> in the peridomicile. In the present work, we reported the first data of prevalence of infection with <em>T. cruzi</em> in the backyard mammals of households in a rural community of Yucatan, Mexico. A total of 84 animals, including swine, sheep, horses, cattle and rabbits, were sampled. Blood samples were collected and processed for DNA extraction of parasite and analyzed by PCR technique. From all animals tested, 75% of the pigs (21/28), 85.71% of the sheep (6/7) and 100% of the horses (8/8), were positive for <em>T. cruzi</em>. Cattle (28) and rabbits (13) were negative. The results obtained constitute an important preliminary evidence to implicate backyard mammals as potential maintaining hosts of <em>T. cruzi </em>in the peridomestic habitat of rural communities in Yucatan.</p>