Toxoplasmosis stands out as a global disease that has felines as definitive hosts. In the municipality of Natal, Rio Grande do Norte State, Brazil, two parks are notable for their ecological and social importance. This study aimed to investigate the presence of Toxoplasma gondii in short hair cats, bats and small non-volant mammals in these two ecological reserves. Altogether, biological samples were obtained from 154 mammals, 92 wild animals from both areas and 62 domestic cats of the Parque da Cidade. In total, 22 (53.7%) non-volant wild mammals, 11 (21.5%) bats and 28 (52.8%) cats were positive for IgG anti-T. gondii antibodies using the Modified Agglutination Test (≥ 25). It was possible to detect the presence of T. gondii DNA, by means of a molecular amplification of a B1 gene fragment (155bp), in 92 tissue samples from wild animals, including Didelphis albiventris, Monodelphis domestica, Artibeus lituratus, Carollia perspicillata and Glossophaga soricina. Of the 62 cats examined by the same molecular method, T. gondii DNA could be detected in 4 cats. In this study, it was observed the circulation of T. gondii in wild species and domestic cats, demonstrating the involvement of wild and domestic animals in the cycle of T. gondii.