The aim of the present study was to investigate antibodies to Toxoplasma gondii in the serum of mules and donkeys bred in the northeast of Brazil. In total, 483 samples were used (395 mules and 88 donkeys) from 4 states (Pernambuco, Rio Grande do Norte, Paraíba, and Sergipe). The indirect immunofluorescence reaction (IFI) technique was used to investigate antibodies to T. gondii with a cut-off point of 64. Positive frequencies of 23.8% and 43.2% were recorded for mules and donkeys, respectively. The state of Pernambuco had the highest prevalence of positive samples (29%) with statistically significant differences for species (P < 0.001) and state (P = 0.048). This is the first study of antibodies to T. gondii in mules and donkeys in these 4 states of the northeastern region of Brazil and serves as a warning to health authorities regarding the risks of ingesting equine meat.