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Neospora caninum is an intracellular parasite which causes abortion in buffalo worldwide. The aim of this study was to determine the seroprevalence of Neospora caninum in buffalo in the province of east-Azerbaijan in northwest Iran. Blood samples were collected from 236 buffalo in the province of east-Azerbaijan for determining the seroprevalence of Neospora caninum. A total of 236 serum samples were tested for anti-neospora antibodies. Serum samples were analyzed for antibodies against N. caninum antigen using a commercial N. caninum ISCOM ELISA kit. Antibodies to N. caninum were found in 42 of the 236 (17.7%) sera based on ELISA test results. This study is the first report of Neospora infection in this area. With regard to seropositivity, no significant difference was observed regarding origin, sex and age (P > 0.05).