Train-wildlife collisions can impact wildlife populations as well as create human and resource management challenges along railways. We identified locations and railroad design features associated with train-wildlife collisions (strikes) on a 134 km section of the Canadian Pacific Railroad (CPR) that travels through the Banff and Yoho National Parks. A 21-year dataset of train strikes with elk (Cervus elaphus), deer (Odocoileus spp.), American black bears (Ursus americanus) and grizzly bears (U. arctos) were compared to relative abundance estimates, and nine train and railroad variables. Train strikes and relative abundance varied spatially for elk, deer and bears. Hotspots and relative risk estimates were used to identify potential problem locations. Hotspots were defined as segments of the train line where strike counts were above the 95% confidence interval based on a Poisson distribution, and could be identified for elk and deer but not bears. Relative risk was estimated as the ratio of strike counts to that expected based on relative abundance. High relative risk locations, where more strikes occurred than were expected, were identified for elk, deer, and bears. Relative abundance was positively correlated with strikes for elk and deer but not bears. Train speed limit was positively associated with strikes for elk and deer. For bears, the number of structures (e.g., overpasses, tunnels, snow sheds and rock cuts) and bridges were positively correlated to strikes. To reduce the risk of train strikes on wildlife, our management recommendations include train speed reduction, habitat modifications and railroad design alterations.