The genus Eustrongylides includes zoonotic nematodes that infect fish species and fish-eating birds of freshwater ecosystems. This study aimed to evaluate the occurrence of Eustrongylides in the paratenic host Perca fluviatilis (European perch) and in the definitive host, Phalacrocorax carbo sinensis (great cormorant), in Lake Annone, a shallow eutrophic lake located in the pre-mountainous area of the Alps in northwest Italy where wintering cormorants coexist with new breeding colonies. A total of 114 European perch and 48 cormorants were examined for the occurrence of Eustrongylides. All parasites collected were identified with microscopic examination and molecular analysis. Overall, 11 specimens of European perch (9.6%) and 13 individuals of cormorants (27%) harbored nematodes identified as fourth-stage larvae and adults of Eustrongylides excisus. The observed prevalence of Eustrongylides spp. appears to be intermediate between the higher values in cormorant breeding areas in northern Europe and the lower prevalence observed in their wintering sites in southernmost Europe. Considering the eutrophication status of freshwater ecosystems and the increasing population of the cormorants, Eustrongylides has an increasing potential range of dispersion in Europe, including Italy; thus an extensive surveillance should be carried out, especially given the zoonotic potential of this nematode.