The Pyrenean desman, a threatened, semiaquatic mammal, is considered a specialist predator feeding on aquatic benthic invertebrates. This categorization comes from visual identification of prey in scat or gut contents, often based on a limited number of samples and locations. We combined diet analyses using next-generation sequencing methods with an extensive survey to explore the summer diet of Pyrenean desmans across the French Pyrenees. This study thus provides an unprecedented level of detail on the trophic ecology of Pyrenean desmans. Our results revealed a diverse diet containing a high proportion of rare prey and substantial consumption of terrestrial prey, which suggests a more generalist diet than previously understood. Three diet groups were identified, with significant differences in prey composition. These differences were not related to geographic location, but rather to local environmental variables. The spatial variation in diet was likely induced by local abiotic parameters that affect prey availability or use of foraging habitats.