Population dynamics of aquatic parasites respond to factors like host availability, habitat age and quality. Amphipods are intermediate hosts for Acanthocephala, a widespread group of parasitic worms. Acanthocephalan infections of amphipods can easily be detected, and the widespread occurrence of amphipods makes their infection status an attractive potential proxy for the ecological status of their aquatic environment, including stressors introduced by urbanization. This study investigated the prevalence and the species-level and genetic diversity of Acanthocephala in the stream amphipod Gammarus fossarum. The study streams cross forested, agricultural and urban landscapes in the eastern foothills of the European Alps. Parasite prevalence ranged from 0% to 8.8% and increased towards downstream reaches independent of surrounding land use. Oxford Nanopore Technology was used to sequence the mitochondrial cytochrome oxidase I barcoding locus to identify parasite species and assess their genetic diversity. The majority of the parasites were Pomphorhynchus tereticollis, which use fish as definitive hosts. Despite their relative abundance in the studied streams, their genetic diversity was low and the most common haplotype was found at all sampling sites, which might indicate population expansion. Amphipods also hosted P. laevis and Polymorphus sp. type 1, the first evidence of this cryptic species within Polymorphus cf. minutus in Austria. Genetic diversity was high in Polymorphus sp. type 1, possibly reflecting a large effective population size due to gene flow maintained by the avian final hosts. The low and downstream-biased prevalence suggests that definitive hosts may be a limiting factor for Acanthocephala populations in small streams.