A cross-sectional study was conducted to assess the effect of neosporosis on productive and reproductive parameters in dairy cows. Cows (n = 2743) from 94 farms located in the most important dairy areas in Costa Rica were used in the study. The size of the herds ranged from 32 to 379 females (mean = 110, median = 125). An indirect ELISA was used to determine the serostatus of the cows towards Neospora caninum. The effect of neosporosis on milk production was analysed by a mixed linear model. In addition, the effects on calving interval (days) and calving to conception interval (days) were analysed by survival analysis. The risk of abortion in relation to N. caninum serostatus was assessed by logistic regression, with herd as a random effect. Overall, 1185 of 2743 cows (43.3%) were seropositive for Neospora. Eighty-nine of 94 (94.7%) farms were classified as Neospora-seropositive. It was estimated that cows seronegative to Neospora produced an additional 84.7 L of milk (P = 0.6). Serostatus did not have a significant effect on the length of the calving interval in the Cox proportional hazard survival analysis (Hazard ratio = 1.2, 95% CI: 0.9, 1.4). The logistic regression model had a weak positive association between serostatus and abortion (OR 1.7, 95% CI: 0.8, 3.9), but did not show a strong association between serostatus and the number of services per conception (OR = 0.95, 95% CI: 0.7, 1.3). In conclusion, there were no significant effects of Neospora serostatus on production and reproductive performance in this study.