Fluctuations in the distribution and abundance of prey resources are an important influence on the foraging ecology of carnivores. Spatio-temporal variation in the diet of river otters (Lontra canadensis), however, is not well understood. In addition, we have limited knowledge about seasonal changes in otter activity at latrine sites and how these changes may relate to changes in otter diet. We used a combination of scat content and stable-isotope analyses to assess the contributions of different prey items to otter diet. We investigated the spatio-temporal variation in the availability of prey groups as it influenced the composition of otter diet and the number of scat deposited at latrine sites. A combination of fish spawning period, water-body type, and lake best described the presence of salmonidae, minnows, and insects in otter scats. The number of scats was best described by a two-week calendar time measurement and geographic location. Scat deposition was positively influenced by a time period when no fish were spawning (early July) and the kokanee (Oncorhynchus nerka) spawning period (early September). In general, the stable-isotope analysis agreed with the results of the scat content analysis: fish dominated the diet, with lesser contributions from other prey items. The stable-isotope analysis, however, suggested that sockeye salmon, larger species of fish (burbot, lake trout), and birds contributed more than was revealed by scat content analysis. Management strategies require accurate and unbiased information on wildlife distribution and abundance that is often measured from surveys of sign; this study provides some of the critical information needed to interpret surveys for river otters. We also suggest implications for other wildlife species.