We studied the frequency of antibodies against Toxoplasma gondii in stray dogs in the city of Oaxaca, Mexico through the evaluation of 154 sera by indirect ELISA. A frequency of 61.7% was found; it was higher in males (45 of 65, 69.2%) than in females (49 of 89, 55.0%), although this difference was not statistically significant. An increase in frequency was observed with age, the lowest being among animals younger than 1 yr (4 of 20, 20.0%) and the highest in dogs older than 7 yr (21 of 25, 84.0%). This is the first study in dogs of this region of Mexico and revealed high T. gondii transmission and evidence of early exposure in animals that are in close contact with contaminated water or raw meat, or both. Further studies are needed in order to understand the role of T. gondii infection in public health.