Turtles face a variety of threats (e.g. habitat destruction, introduced predators) that are pushing many species towards extinction. Vehicle collisions are one of the main causes of mortality of adult freshwater turtles. To conceptualise the level of threat that roads pose to Australians turtles, we analysed data gathered through the citizen science project TurtleSAT along the Murray River. We recorded 124 occurrences of turtle road mortality, which included all three local species (Chelodina expansa, Chelodina longicollis, and Emydura macquarii). Chelodina longicollis was the most commonly reported species killed on roads. We found that rain and time of year affect the likelihood of C. longicollis being killed on roads: increased turtle mortality is associated with rain events and is highest during the month of November, which coincides with their nesting season. Chelodina longicollis was most likely to be killed on the Hume Highway and roads around major urban centres; therefore, we recommend that governing bodies focus management practices and increase awareness at these locations. The degree of road mortality that we detected in this study requires mitigation, as it may contribute to the decline of C. longicollis along the Murray River.