This is the first report on the presence of acanthocephalan parasite Echinorhynchus truttae in brown trout (Salmo trutta) from Spain. A total of 343 fish were captured by local anglers from 19 rivers in the adjacent Tambre and Ulla watersheds in Galicia (north-western Spain). Macroscopic and microscopic analyses of the intestinal contents revealed the presence of adults and/or eggs of E. truttae in 53 of the 123 trout from Tambre river basin (43.1%). By contrast, parasitic forms of this acanthocephalan were only observed in eight of the 220 fish from Ulla basin (3.6%), showing significant differences between the prevalences obtained in two adjacent watersheds (P < 0.001, odds ratio 19.0). Prevalence was significantly higher in specimens >3 years (length >26.0 cm) than in younger specimens (P < 0.05). The absence of Gammarus pulex in the region suggests that native gammarid species in Galicia (Echinogammarus lusitanicus and Echinogammarus beriyoni) may act as intermediate host in the life cycle of E. truttae. Different prevalences of E. truttae indicate that the presence/abundance of the intermediate crustacean host may be different in the two river basins, probably as a consequence of various abiotic factors and anthropogenic activities.