Neosporosis is a parasitic disease caused by the protozoan Neospora caninum, which causes economic losses due to reproductive problems. In order to detection of antibodies against N. caninum in buffaloes from Colon Municipality in Zulia State, Venezuela, determinating the seroprevalence and analyzed the risk factors associated with this disease. One hundred seventy four samples of blood serum using ELISA test were processed. Detected antibodies against N. caninum showing a seroprevalence of 30.4%, 100% of farms were seropositive, animals and statistical analysis indicated that there had not association between seropositivity and the type of canine food, veterinary care, type of mounting and biological waste management on a farm. These results indicate a moderate exposure of water buffaloes to the protozoan in the studied area.