Parasites with complex life-cycles and trophic transmission are expected to show low specificity towards final hosts. However, testing this hypothesis may be hampered by low taxonomic resolution, particularly in helminths. We investigated this issue using two intestinal fish parasites with similar life-cycles and occurring in sympatry, Pomphorhynchus laevis and Pomphorhynchus tereticollis (Acanthocephala). We used species-specific ITS1 length polymorphism to discriminate parasite species from 910 adult acanthocephalans collected in 174 individual hosts from 12 fish species. Both P. laevis and P. tereticollis exhibited restricted host range within the community of available fish host species, and transmission bias compared to their relative abundance in intermediate hosts. The two parasites also exhibited low niche overlap, primarily due to their contrasting use of bentho-pelagic (P. laevis) and benthic (P. tereticollis) fish. Furthermore, parasite prevalence in intermediate hosts appeared to increase with taxonomic specificity in definitive host use. Comparison of P. laevis and P. tereticollis adult size in the two main definitive hosts, barbel and chub, suggested lower compatibility towards the fish species with the lowest parasite abundance, in particular in P. laevis. The determinants of low niche overlap between these two sympatric acanthocephalan species, and the contribution of definitive host range diversity to parasite transmission success, are discussed.