Neospora caninum, an apicomplexan protozoan parasite, is recognized as a major cause of abortion in cattle. However, limited information is presently available on the seroprevalence of Neospora antibodies in horses worldwide. The aim of the present study is to determine serological prevalence of Neospora infection in horses in Iran. Blood samples were obtained from 200 horses and tested for serum antibodies against Neospora spp. by the Neospora modified direct agglutination test (N-MAT). Antibodies were found in 64 (32%) horses being tested with titers of 1:80. This is the first serological survey for Neospora antibodies performed on horses in Iran.