The aim of this study was to assess the prevalence of Toxoplasma gondii infection in rodents that coexist with ocelots in north-eastern Mexico. Eighty rodents of five genera were captured and their serum samples tested for specific IgG antibodies to T. gondii by in-house indirect ELISA using three different conjugates. Prevalences of 7% (3/44) and 33% (4/12) were found in Sigmodon hispidus and Liomys irroratus, respectively, and were significantly different. All Baiomys taylori and Oligoryzomys fulvescens were negative for the presence of anti-T. gondii IgG antibodies. The samples from Peromyscus spp. could not be analyzed because none of the three conjugates tested recognized their immunoglobulins. Infection was confirmed in one single specimen of L. irroratus by qPCR, which generated an estimate of 146 parasites per mg of muscle tissue. The results strongly support the notion of active T. gondii transmission between rodents and felines in this zone of Mexico and an important role of some rodent species in the sylvatic cycle of T. gondii.