The seroprevalence and risk factors for exposure to Neospora caninum and Neospora hughesi in Ontario broodmares were investigated. The prevalence estimates for N. caninum and N. hughesi were 27.4% (60/219) and 29.7% (65/219), respectively. The IFAT cut-off titre for N. caninum and N. hughesi was 1:40 and 1:160, respectively. Risk factors for N. caninum included presence of farm dogs (OR = 6.70; 95% CI = 2.14 – 20.97; p = 0.001), and high stocking density (OR = 2.83; 95% CI = 1.27 – 6.30; p = 0.011). Presence of livestock, excluding cattle, was associated with reduced risk of exposure (OR = 0.17; 95% CI = 0.05 – 0.53; p = 0.002). The only risk factor for exposure to N. hughesi was feeding hay on the ground in the paddock (OR = 4.31; 95% CI = 1.65 – 11.22; p = 0.003). This study demonstrated significant exposure to Neospora spp. in Ontario broodmares.