Toxoplasmosis is a parasitic disease resulting from infection with the apicomplexan Toxoplasma gondii (Nicolle et Manceaux, 1908), one of the world's most common parasites in warm-blooded animals, including humans. Sources of infection can be exposed to infected cat faeces, mother-to-child transmission during pregnancy, and notably, the consumption of undercooked contaminated meat of intermediate hosts. In Thailand, water buffaloes are highly valued for rice cultivation, traditional culture and meat production. Like several other mammalian species, these animals play a role as reservoirs of T. gondii, thus representing a threat to human health. The seroprevalence of T. gondii infection in swamp buffaloes was examined in southern and northeastern Thailand. In total, serum samples of 721 water buffaloes were collected from seven provinces (Ubon Ratchathani, Roi Et, Si Sa Ket, Surin, Buri Ram, Sakon Nakhon, and Songkhla) and examined for the presence of T. gondii infection using commercial latex agglutination test kits (TOXOCHECK-MT, Eiken Chemical Co., Tokyo, Japan). Of the 721 animals analysed, 49 (6.8%) were positive for T. gondii. Songkhla province had the highest seroprevalence (14.7%) among the seven provinces covered in this survey. There was a potential risk to local citizens of T. gondii infection identified by the present study, notably in northeastern Thailand, where despite lower seroprevalence consuming raw buffalo meat salad should be restricted to avoid the risk of zoonotic infections.