Since being successfully reintroduced into Ontario, Canada, wild turkey (Meleagris gallopavo) populations have undergone robust growth and range expansion. This, along with increases in land use changes from human population growth and subsequent developments in agriculture and livestock production, has heightened opportunities for interactions between wild turkeys, domestic poultry, and humans. As conspecifics, wild and domestic turkeys are susceptible to infection and disease from many of the same pathogens. Thus, transmission by direct or indirect contact is a potential health threat to both groups, particularly with the overlapping range of wild turkeys in Ontario with numerous commercial and backyard poultry operations. However, these threats are difficult to assess due to knowledge gaps in the prevalence and geographic distribution of potential pathogens circulating among wild turkeys. We assessed for potentially pathogenic bacteria in free-ranging, hunter-harvested wild turkeys in Ontario (n = 152) by cloacal swab culture for Campylobacter spp., Salmonella spp., and Escherichia coli and culture of lung and spleen for Pasteurella multocida, Ornithobacterium rhinotracheale, and Erysipelothrix rhusiopathiae. Antimicrobial resistance testing was also performed on E. coli isolates. Generic E. coli isolates were recovered from 69.1% (105/152) of wild turkeys tested, and two (1.9%) of these isolates exhibited resistance to azithromycin and one (1.0%) to ampicillin. Intermediate susceptibility to chloramphenicol was observed in one (1.0%) isolate. One (0.7%) wild turkey swab tested positive for C. jejuni, but no samples were positive for P. multocida, Salmonella spp., O. rhinotracheale, or E. rhusiopathiae. To our knowledge, this is the first survey of these bacteria and assessment for antimicrobial resistance among wild turkeys in Ontario.