the ojibway Prairie Complex in Windsor contains the largest protected tallgrass prairie ecosystem in ontario and supports numerous species at risk. Despite its ecological significance, it is crossed by multiple high-traffic roads. Road mortality is a major threat to endangered species in Canada, particularly reptiles. the main goal of this study was to describe the nature and extent of vertebrate road mortality, with a focus on reptiles, on roads bisecting the ojibway Prairie Complex, and the Greater Park ecosystem, in Windsor and Lasalle, ontario. a systematic road mortality survey was conducted by bicycle along seven roads (12.5 km) in 2010, 2012, and 2013. also, opportunistic observations (n = 103) spanning over 30 years were assembled from a variety of sources. in total, 2083 vertebrates (49 species), including 446 reptiles (11 species), were recorded "dead on road" during systematic surveys. the highest diversity of reptiles was recorded on Matchette Road, whereas the highest rate of reptile mortality was recorded on Malden Road. Reptile species at risk were killed on all roads surveyed. Combining systematic and opportunistic data, we found seven reptile species at risk: Butler's Gartersnake (Thamnophis butleri), eastern Foxsnake (Pantherophis vulpinus), eastern Massasauga (Sistrurus catenatus catenatus), Blanding's turtle (Emydoidea blandingii), eastern Musk turtle (Sternotherus odoratus), northern Map turtle (Graptemys geographica), and snapping turtle (Chelydra serpentina). Reptile road mortality "hotspots" occurred where each road is intersected by a naturalized utility right-of-way. our results can be used to focus mitigation efforts in space and time to reduce mortality rates and enhance connectivity in the ojibway Prairie Complex and Greater Park ecosystem.