A seroepidemiological survey of Neospora caninum infection in dairy cattle was carried out in China's southern Guangdong Province between July 2009 and March 2010. A total of 370 serum samples of dairy cattle was collected from 5 farms and examined for antibodies to N. caninum by enzyme-linked immunosorbent assay. The overall seroprevalence of N. caninum in dairy cattle was 18.9% (70/370). The seroprevalence of N. caninum in aborting cows (22.7%) was higher than that in nonaborting cows (16.3%), but the difference was not statistically significant (P > 0.05). Five-yr-old dairy cattle had the highest seroprevalence (27.8%), followed by those that were 6-yr-old (20.4%). Dairy cattle with 4 pregnancies had the highest seroprevalence (29.2%). There was no apparent association of N. caninum seropositivity with age or number of pregnancies (P > 0.05). The results of the present survey indicated that the infection with N. caninum is prevalent in dairy cattle of all ages in southern China, which may be one of the causes of bovine abortion. This is the first report of seroprevalence of N. caninum in dairy cattle in southern China.