A cross-sectional study was undertaken (October 2010 to August 2011) to estimate the prevalence of paratuberculosis in the small ruminant dairy industries in Ontario, Canada. Blood and feces were sampled from 580 goats and 397 sheep (lactating and 2 y of age or older) that were randomly selected from 29 randomly selected dairy goat herds and 21 convenience-selected dairy sheep flocks. Fecal samples were analyzed using bacterial culture (BD BACTEC MGIT 960) and polymerase chain reaction (Tetracore); serum samples were tested with the Prionics Parachek enzyme-linked immunosorbent assay (ELISA). Using 3-test latent class Bayesian models, true farm-level prevalence was estimated to be 83.0% [95% probability interval (PI): 62.6% to 98.1%] for dairy goats and 66.8% (95% PI: 41.6% to 91.4%) for dairy sheep. The within-farm true prevalence for dairy goats was 35.2% (95% PI: 23.0% to 49.8%) and for dairy sheep was 48.3% (95% PI: 27.6% to 74.3%). These data indicate that a paratuberculosis control program for small ruminants is needed in Ontario.