The aim of the present study was to determine and to compare through an indirect enzyme linked immunosorbent assay (ELISA) test, the presence of anti-Neospora caninum antibodies in city and farm dogs, as well as in farm cows, and the relationship among them. The correlation between anti-N. caninum antibodies in farm dogs and cattle was also assessed. The research was conducted in the dairy region of Tizayuca, Hidalgo, Mexico. The frequency of anti-N. caninum antibodies was significantly higher in farm dogs (n= 14)(51%) when compared to those from the city (n= 6)(20%)(P< 0.05), suggesting that farm dogs have a higher risk of exposure to the parasite. There was no significant difference in seropositivity between males (n= 11)(39%) and females (n= 9)(33%)(P> 0.05). The frequency of anti-N. caninum antibodies in farm cattle was significantly higher in farms with dogs (n= 158)(58%) when compared to those with no dogs (n= 43)(35%)(P< 0.05). These results suggest the possible transmission of the parasite from dogs to cattle.