Following extirpation from Ontario, Canada in the early 1900s, Eastern Wild Turkeys (EWTs; Meleagris gallopavo silvestris) were successfully reintroduced to the province in 1984. Despite the subsequent establishment of robust populations and biannual hunting seasons, data on the circulation of potential pathogens in these birds are lacking. Similarly, the interface between EWTs and poultry is poorly understood and includes possible bidirectional pathogen transmission via direct or indirect contact. Mycoplasma and Eimeria spp. are potential pathogens in Galliformes, and our objective was to determine their prevalence and distribution in Ontario EWTs. During the 2015 spring hunting season (April and May), oropharyngeal swabs from 147 hunter-harvested and five opportunistically collected EWTs from southern Ontario were cultured for Mycoplasma spp. The intestinal or cloacal contents of 107 of these birds and an additional 24 opportunistically and biologistcollected EWTs were analyzed for Eimeria spp. using PCR or fecal flotation. At least one Mycoplasma spp. was isolated from 98.7% (150/152) of EWTs, with six species identified. Mycoplasma gallopavonis was identified most commonly in 96.7% (147/152), followed by Mycoplasma gallinaceum in 23.7% (36/152). Potential poultry pathogens (Mycoplasma meleagridis, Mycoplasma iowae, and Mycoplasma synoviae) were isolated from swabs of five (3.3%) EWTs. Coinfections with up to three Mycoplasma spp. were detected in 36.8% (56/152) of EWTs. Most EWTs tested positive for Eimeria spp. oocysts (75.6%; 99/131). A subset of positive samples (n=16) were characterized by PCR, which detected the following species: Eimeria meleagrimitis (93.8%), Eimeria adenoeides (93.8%), Eimeria gallopavonis (56.3%), and Eimeria meleagridis (12.5%). The majority (93.8%) of these samples were positive for more than one Eimeria spp. We showed that numerous, mostly nonpathogenic Mycoplasma and Eimeria spp. circulate in EWTs across southern Ontario, and this helped to establish baseline information for comparison with future surveillance and monitoring.