Complete sequences of ribosomal and mitochondrial genes of the giant liver fluke Fascioloides magna are presented. In particular, small subunit (18S) and internal transcribed spacers (ITS1 and ITS2) of the ribosomal gene (rDNA), as well as cytochrome c oxidase subunit I (cox1) and nicotinamide dehydrogenase subunit I (nad1) of the mitochondrial DNA (mtDNA), were analyzed. The 18S and ITS sequences were compared with previously published sequences of the liver fluke Fasciola hepatica. Fixed interspecific genetic differences were determined that allow molecular differentiation of F. magna and F. hepatica using either the PCR-RFLP method or PCR amplification of species-specific DNA regions. Additionally, intraspecific sequence polymorphism of the complete cox1 and nad1 mitochondrial genes in geographically distinct F. magna populations was determined. Based on the sequence divergences, short (< 500 bp) variable regions suitable for broader biogeographical studies of giant liver fluke were designed.

The species-specific ribosomal internal transcribed spacer 2 (ITS2) markers were designed for PCR-based molecular differentiation of Fasciola hepatica, Fascioloides magna, Dicrocoelium dendriticum and Paramphistomum cervi, liver and stomach flukes of domestic and free living ruminants. Complete ITS2 sequences were obtained for D. dendriticum and P. cervi, for the later species, ITS2 structure was determined for the first time. Intraspecific variation within geographically distant populations was found to be either very low (F. hepatica; D. dendriticum) or even absent (F. magna; P. cervi). ITS2 regions with the absence of intraspecific polymorphisms but with interspecific sequence heterogeneity were applied for design of speciesspecific primers. The specificity of developed primers was tested on genomic DNA isolated from adult individuals of studied fluke species. Application of the primers is of particular value for molecular differentiation of morphologically hardly distinguishable F. hepatica, F. magna and P. cervi eggs after coprological examinations.

Pomphorhynchus tereticollis (Rudolphi, 1809) is here redescribed on the basis of Rudolphi’s material, deposited in the Museum für Naturkunde Berlin, and on acanthocephalans recently collected from the type host Platichthys flessus (L.) and the region embodying the type locality. Out of the paratypes of P. tereticollis, the lectotype and paralectotypes have been designated. Their morphology fits well with that of newly collected material of P. tereticollis dissected from the type fish host from the Baltic coast near Stralsund. The resurrection of P. tereticollis, previously considered a synonym of Pomphorhynchus laevis (Zoega in Müller, 1779), is supported by several morphological features distinguishing the two Pomphorhynchus species: 1. The basal parts of the proboscis hooks located on the posterior proboscis half possess proximal projections in P. tereticollis but not in P. laevis. This shape of the hook bases is clearly visible only in unfixed fresh worms; 2. The last hooks are situated at the anterior part of the bulbus or rarely at the posterior-most end of the proboscis in P. tereticollis, while they lie anterior to the end of the proboscis in P. laevis; 3. The proboscis hooks No. 5 or 6 are markedly stout (robust) and clearly distinct in comparison with the surrounding hooks in P. tereticollis, while less robust and more similar to the hooks in P. laevis. In addition, genetic divergence between P. tereticollis and P. laevis based on ITS1, ITS2 and COI sequencing supports the existence of two distinct species and reveals that some isolates previously identified as P. laevis were actually P. tereticollis. Previous and present morphological and genetic data show that both Pomphorhynchus species occur in freshwaters throughout Europe and may infect the same fish hosts, such as chub and barbel, and also several species of isopods (Gammaridae). This study also provides morphological evidence that Pomphorhynchus intermedius Engelbrecht, 1957 is a synonym of P. tereticollis, because the only discrimination character of the former species, the “existing but small proximal projections of basal parts of the proboscis hooks located on the posterior proboscis half” are present also in P. tereticollis.