In this study, serum samples of 53 wild animals from two different states from the southeast region of Brazil were analyzed for the presence of anti–Toxoplasma gondii antibodies by the modified agglutination test (MAT), with a cut-off of 1 : 5 for birds and of 1 : 25 for mammals. Out of the sampled animals, 27 were birds and 26 were mammals, and from this total, 83% (n = 44) were free-living animals. Antibodies were found in 13 mammals, from which 11 were free-living animals, and in five birds, all of which were free-living. In this study, T. gondii antibodies were detected in four bird species (crested seriema, Cariama cristata; buff-necked ibis, Theristicus caudatus; picazuro pigeon, Patagioenas picazuro; and burrowing owl, Athene cunicularia) and in a giant anteater (Myrmecophaga tridactyla) for the first time.