Diet interactions between native and non-native fishes may influence the establishment of native species within their historical range (i.e., reintroduction). Therefore, we illustrated the food web structure of and followed the transition of the federally endangered humpback chub Gila cypha into a novel food web following translocation and determined the potential for a non-native species, rainbow trout Oncorhynchus mykiss, to influence translocation success. Humpback chub and rainbow trout used resources high in the food web and assimilated similar proportions of native fishes, suggesting non-native rainbow trout may occupy an ecological role similar to humpback chub. Subsequently, humpback chub may be well suited to colonise tributaries because of their ability to consume resources high in the food web. Additionally, diet partitioning may occur between all members of the fish community as indicated by separation in trophic niche space and little trophic overlap; although all species, particularly bluehead sucker Catostomus discobolus, used a broad range of food resources. Rainbow trout stomach content analysis corroborated stable isotope analysis and suggested rainbow trout diet consisted of aquatic and terrestrial macroinvertebrates, while larger rainbow trout (\u3e120 mm total length) consumed a greater proportion of fish (incidence of piscivory = 5.3%). Trophic interactions may reveal an underutilized niche space or biotic resistance to the establishment of translocated native fishes. Continued translocation of humpback chub into tributaries appears to be one option for conservation. However, successful establishment of humpback chub may depend on continued removal of non-native trout, increasing availability of diet sources at higher trophic levels
