Roads have considerable ecological effects that threaten the survival of some species, including many terrestrial carnivores. The western polecat is a small-medium sized mustelid native to Asia and Europe, including Britain where its historical stronghold is in Wales. Polecats are frequently killed on roads and road casualties represent the most common source of data on the species in the UK. However, little is known about the factors that increase the risk of collision. We used Generalized Linear Mixed Models to explore seasonal patterns in collisions as well as using Principal Component Analysis and regression modelling to identify landscape characteristics associated with polecat road casualties in Wales. Polecat road casualties had a bimodal distribution, occurring most frequently in March and October. Casualties were more frequently associated with road density, traffic volume, presence of rabbits, habitat patchiness and the abundance of proximal improved grassland habitat. Casualties were negatively associated with elevation and the abundance of semi-natural grassland habitat. The results of this study provide a framework for understanding and mitigating the impacts of roads on polecats in their historic stronghold, hence has considerable value to polecat conservation as well as broader applicability to ecologically similar species.