Cercariae (Trematoda, Digenea) in European freshwater snails - a checklist of records from over one hundred years

Freshwater snails of the family Lymnaeidae are of a great parasitological importance because of the very numerous helminth species they transmit, mainly trematodiases of large medical and veterinary impact. The present knowledge on the genetics of lymnaeids and on their parasite-host inter-relationships is far from being sufficient. The family is immersed in a systematic-taxonomic confusion. The necessity for a tool which enables species distinction and population characterization is evident. This paper aims to review the European Lymnaeidae basing on the second internal transcribed spacer ITS-2 of the nuclear ribosomal DNA. The ITS-2 sequences of 66 populations of 13 European and 1 North American lymnaeid species, including the five generic (or subgeneric) taxa Lymnaea sensu stricto, Stagnicola, Omphiscola, Radix and Galba, have been obtained. The ITS-2 proves to be a useful marker for resolving supraspecific, specific and population relationships in Lymnaeidae. Three different groupings according to their ITS-2 length could be distinguished: Radix and Galba may be considered the oldest taxa (370-406 bp lengths), and Lymnaea s. str., European Stagnicola and Omphiscola (468-491 bp lengths) the most recent, American Stagnicola and Hinkleyia being intermediate (434-450 bp lengths). This hypothesis agrees with the phylogeny of lymnaeids based on palaeontological data, chromosome numbers and radular dentition. ITS-2 sequences present a conserved central region flanked by two variable lateral regions corresponding to the 5' and 3' ends. The number of repeats of two microsatellites found in this conserved central region allows to differentiate Radix from all other lymnaeids. Phylogenetic trees showed four clades: (A) Lymnaea s. str., European Stagnicola and Omphiscola; (B) Radix species; (C) Galba truncatula; and (D) North American stagnicolines. ITS-2 results suggest that retaining Stagnicola as a subgenus of Lymnaea may be the most appropriate and that genus status for Omphiscola is justified. Radix shows a complexity suggesting different evolutionary lines, whereas G. truncatula appears to be very homogeneous. North American and European stagnicolines do not belong to the same supraspecific taxon; the genus Hinkleyia may be used for the American stagnicolines. Genetic distances and sequence differences allowed us to distinguish the upper limit to be expected within a single species and to how different sister species may be. S. palustris, S. fuscus and S. corvus proved to be valid species, but S. turricula may not be considered a species independent from S. palustris. Marked nucleotide divergences and genetic distances detected between different S. fuscus populations may be interpreted as a process of geographic differentiation developping in the present. Among Radix, six valid species could be distinguished: R. auricularia, R. ampla, R. peregra (=R. ovata;=R. balthica), R. labiata, R. lagotis and Radix sp. The information which the ITS-2 marker furnishes is of applied interest concerning the molluscan host specificity of the different trematode species. The phylogenetic trees inferred from the ITS-2 sequences are able to differentiate between lymnaeids transmitting and those non-transmitting fasciolids, as well as between those transmitting F. hepatica and those transmitting F. gigantica. The Fasciola specificity is linked to the two oldest genera which moreover cluster together in the phylogenetic trees, suggesting an origin of the Fasciola ancestors related to the origin of this branch. European Trichobilharzia species causing human dermatitis are transmitted only by lymnaeids of the Radix and Lymnaea s. str.-Stagnicola groups. Results suggest the convenience of reinvestigating compatibility differences after accurate lymnaeid species classification by ITS-2 sequencing. Similarly, ITS-2 sequencing would allow a step forward in the appropriate rearrangement of the actual systematic confusion among echinostomatids.

A survey of the larval stages (cercariae and metacercariae) of trematodes (Digenea) found in planorbid snails in Central Europe (Austria, Czech Republic, south-east Germany, Hungary, Poland and the Slovak Republic) is presented based on a study of 7,628 snails of 12 species examined between 1998-2006. A total of 34 trematode larval stages, comprising cercariae of 28 species and metacercariae of seven species (one species occurred both as cercaria and metacercaria) of nine families were found in 898 (11.5%) snails of eight species. The dominant cercariae were those belonging to the Rubenstrema exasperatum (Rudolphi, 1819)/Neoglyphe locellus (Kossack, 1910) species complex, Tylodelphys excavata (Rudolphi, 1803) and Echinostoma spiniferum (La Valette, 1855) sensu Nasincová (1992), all from Planorbarius corneus (Linnaeus). Almost the same spectrum of cercariae of the families Echinostomatidae, Plagiorchiidae and Omphalometridae was found in the present study as in previous reports; however, a considerably lower spectrum of cercariae of the families Diplostomidae and Strigeidae was recorded. The most frequent metacercariae were those of Echinoparyphium aconiatum Dietz, 1909, Neoglyphe locellus and Moliniella anceps (Molin, 1859), all occurring mainly in P. corneus. The most heavily infected snail species was P. corneus, followed by Planorbis planorbis (Linnaeus) and Segmentina nitida (Müller). The widest spectrum of trematode species was found in P. planorbis and P. corneus. Forty-two cercariae identified to the species level belonging to 15 families, plus an additional 43 taxa recorded under generic or provisional names, were reported from 11 species of planorbids in previous studies carried out in Central Europe. However, the actual number of trematode species occurring in the planorbid snails is probably much lower, because many, if not most, larval stages reported under provisional names or unidentified to the species level may be conspecific with identified adult forms. A key to the cercariae and metacercariae recorded from planorbids in Central Europe, together with illustrations of those species encountered most frequently in the field, is provided to facilitate identification.