The successful reintroduction of Wild Turkeys (Meleagris gallopavo) to Ontario, Canada, has led to established populations in southern portions of the province and currently allows for biannual hunting seasons. These populations geographically overlap Domestic Turkey farms, an important sector of the provincial agri-food industry. Potential pathogen transmission between Wild Turkeys and Domestic Turkeys (Meleagris gallopavo) is a concern, because they are susceptible to infection with many of the same pathogens and have direct and indirect contact in outdoor or open farm settings and contaminated environmental substrates. However, data concerning potential poultry pathogens in Wild Turkeys in Canada are scarce. Thus, we assessed the prevalence and geographic distribution of geographically relevant viruses in Ontario Wild Turkeys. Oropharyngeal and cloacal swabs were tested for avian influenza viruses (AIV) by real-time reverse transcriptase (RT)-PCR (n=207), pooled tissues for lymphoproliferative disease virus (LPDV; n=183) and reticuloendotheliosis virus (n=119) by PCR, and gross skin lesions by real-time RT-PCR for avian poxvirus (n=8). We sequenced a fragment of the gag polyprotein (p31) gene of LPDV on a subset (n=10) of LPDV-positive samples for phylogenetic analysis and tested additional upland game bird species (n=39) and domestic fowl for LPDV (n=17). To the best of our knowledge, we document the first detection of LPDV in Wild Turkeys in Canada, with a prevalence of 65% (119/183). Phylogenetic analysis revealed that LPDV sequences from Ontario were genetically similar to other North American strains and did not group into separate clades. Reticuloendotheliosis virus was detected in 4% (5/119) of LPDV-positive Wild Turkeys. Grossly evident skin lesions from five Wild Turkeys tested positive for poxvirus, and all turkeys tested negative for AIV. This study provides evidence of LPDV circulation in Canada and provides a baseline for comparison with future Wild Turkey pathogen surveillance and monitoring in Ontario and elsewhere.