The aim of the present study was to determine the seroprevalence of Toxoplasma gondii in horses used for sporting purposes in the Province of Riyadh, Saudi Arabia. In total, 266 serum samples from clinically healthy horses were analyzed for anti–T. gondii antibodies using the Sabin-Feldman dye test. Antibodies to T. gondii were found in 84 (31.6%) horses, with specific titers of 1∶16 (78 with a prevalence of 29.3%), 1∶64 (4 with a prevalence of 1.5%), and 1∶256 (2 with a prevalence of 0.8%). The number of seropositive horses in Shaqra (43.7%) was considerably higher than in other regions, with the lowest occurring in Al-Majmaah (11.0%). However, the differences in seroprevalence between the 6 locations were not statistically significant (P > 0.05). In contrast, the overall seroprevalence in Riyadh province was higher than that reported in other countries. Horses are not a direct source of infection for human toxoplasmosis in this region. Usually human infection is facilitated via cats, which are the reservoir hosts for this parasite.