Dairy industry is growing in Nepal and western dairy pocket area is the main milk producing area in Chitwan district. This study was carried out to determine the seroprevalence and risk factors associated with N. caninum in western dairy pocket area in Chitwan district of Nepal. A cross-sectional study was conducted from April 2014 to July 2014 among the small holder dairy farms, and out of 2188 animals from different sampling villages, 186 (8.5%) animals were randomly selected. Individual blood samples (5-10 ml) were collected and screened for N. caninum antibodies by ELISA test kit. The study showed that apparent overall prevalence of N. caninum to be 4.84% and true prevalence, 4.07%. The prevalence varied from a low of 0% to a high of 13.16% in various VDCs. The prevalence rates among Holstein-Friesian cross and Jersey cross Cattle were 6.94% and 3.51% respectively. The seroprevalence of N. caninum was found to be 16.13% and 2.5% in animals with and without history of abortion respectively, showed statistically significant different (p value <0.05). Similarly, prevalence were 13.64% and 3.66% in animals with and without presence of dog respectively and 8.5%, 3.39% and 0% in the animals of age group of 3-5 years, 1-3 years and above 5 years respectively, showed statistically non-significant association (p value <0.05). This study shows that N. caninum is associated with abortion in dairy cattle of Chitwan region. The study may contribute the base line data of N. caninum in Nepal for future preventive strategy for stake-holders and government.