A cross-sectional study was carried out to determine the seroprevalence and to identify risk factors associated with Neospora caninum infection in 62 dairy herds (n = 671 cows) in Jordan between January and June 2007. Information regarding herd management was recorded through personal interviews with farmers. Antibodies against N. caninum were detected using an indirect ELISA test. Chi-square analysis and multivariable logistic regression model were used to identify risk factors associated with N. caninum seropositivity. The true prevalence of antibodies against N. caninum in individual cows and cattle herds was 35 and 66.5 %, respectively. There was no significant difference in the percentage of seropositive animals between different age groups. Multivariable logistic regression model revealed workers frequently visiting nearby farms as a risk factor for seropositivity to N. caninum, while presence of a calving pen was suggested as a protective factor. Amman, Balqa, and Mafrak governorates had significantly lower seroprevalence to N. caninum compared to other Jordanian governorates. Results of this study indicated that N. caninum infection may be widespread in Jordan.