The increasing consumption of fish has raised public health concerns regarding fish-borne zoonotic helminths (FBZHs), which are recognized as significant food-borne parasites worldwide. In freshwater environments, Clinostomum complanatum, Opisthorchis felineus, Pseudamphistomum truncatum, Dibothriocephalus latus and Eustrongylides excisus are of particular concern in Italy and neighbouring countries. This study aimed to assess the prevalence of these FBZHs in five commercially and ecologically relevant freshwater fish species from Lake Varese, a heavily anthropized and understudied basin in northern Italy. A total of 59 fish were examined via necropsy and stereomicroscopic inspection of skeletal muscles. Only Eustrongylides spp. larvae were detected, with a prevalence of 16.9%. Molecular analysis (ITS region) identified them as E. excisus. This study reports, for the first time in Western Europe, E. excisus in Sander lucioperca (p = 12.5%) and Esox lucius (p = 8.3%). The highest prevalence occurred in Silurus glanis (p = 37.5%), followed by Perca fluviatilis (p = 25.0%), while Tinca tinca showed no infection. These findings confirm that among the FBZHs considered, E. excisus is currently present and expanding both in host range and geography in Italian lakes, underscoring the need for updated epidemiological data to support risk assessment, food safety and zoonotic parasite control in freshwater fisheries.