The frequency of Toxoplasma gondii antibodies in stray and household dogs in Guangzhou, China was examined by ELISA on serum samples from 150 animals (36 strays and 114 from households) and the overall prevalence was 21.3%. The extent of infection in stray dogs (33.3%) was significantly higher (P < 0.05) than in household dogs (17.5%). Infection in male and female dogs of both groups was not significantly different (P ≥ 0.05), i.e., 31.8% versus 35.7% for male and female in stray dogs, and 14.5% versus 22.2% in household dogs. The results of the present investigation indicate that the seroprevalence of T. gondii infection in dogs was high in Guangzhou, especially in strays. Therefore, it is essential to implement integrated strategies to prevent and control T. gondii infection in both stray and household dogs.