Host specificity and geographical distribution of Eubothrium in European salmonid fish

The host specificity and distribution of Eubothrium crassum (Bloch, 1779) and Eubothrium salvelini (Schrank, 1790), morphologically fairly similar pseudophyllidean tapeworms parasitizing salmonid fish, were critically assessed on the basis of morphological and genetic evaluation of extensive material collected from different definitive hosts and geographical regions in Europe. Eubothrium crassum occurs in fish of the genera Salmo, i.e. salmon (S. salar - both freshwater and marine), sea trout (S. trutta trutta), brown trout (S. trutta fario), and lake trout (S. trutta lacustris), and also in Danubian salmon (Hucho hucho) and vendace (Coregonus albula). Eubothrium salvelini parasitizes Arctic char (Salvelinus alpinus) and brook trout (Salvelinus fontinalis) in Europe, and also whitefish (Coregonus wartmanni). Rainbow trout (Oncorhynchus mykiss), which is not a native European fish species, was found to be a suitable definitive host for both Eubothrium species, which may occur simultaneously in the same fish. Previous records of E. crassum in Arctic char and brook trout, and those of E. salvelini in fish of the genus Salmo were most probably misidentifications. Most studies of Eubothrium have involved salmonids from the northern part of Europe, with few records from southern and south-eastern Europe. This study also confirmed the reliability of the morphology of the apical disc for the discrimination of E. crassum and E. salvelini

New data on the morphology of Comephoronema oschmarini (Nematoda, Cystidicolidae), a little-known gastrointestinal parasite of Lota lota (Teleostei) in Palaearctic Eurasia

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Redescription of Eubothrium fragile (Rudolphi, 1802) and E. rugosum (Batsch, 1786) (Cestoda: Pseudophyllidea), parasites of fish in the Holarctic Region

Two fish cestodes, the little-known Eubothrium fragile (Rudolphi, 1802) and E. rugosum (Batsch, 1786), the type species of the genus Eubothrium Nybelin, 1922, are redescribed on the basis of new material from twaite shad, Alosa fallax (Lacépède, 1803), from England and burbot, Lota lota (Linnaeus, 1758), from Russia, respectively. The tapeworms are compared with two other species of the genus, E. crassum (Bloch, 1779) and E. salvelini (Schrank, 1790), common parasites of salmonid fish in the Holarctic. The most notable differential characters are the size and the shape of the scolex (smaller and oval in E. fragile), the shape of the apical disc (four or more indentations in E. crassum), the number and size of the testes (the largest and least numerous in E. rugosum), and the position and size of the vitelline follicles (almost entirely cortical in distribution in E. fragile and E. crassum versus largely medullary in E. rugosum and E. salvelini). A comparison of species has also shown the morphological similarity of the freshwater species (E. rugosum and E. salvelini) on one hand and those of marine origin, E. fragile and E. crassum, on the other, with the latter species occurring also in fresh waters. A key to the identification of the species studied is also provided

Morphometric analysis of four species of Eubothrium (Cestoda: Pseudophyllidea) parasites of salmonid fish: An interspecific and intraspecific comparison

Four species of the genus Eubothrium (E. crassum, E. fragile, E. rugosum and E. salvelini) were subjected to morphometric comparison. Discriminant analysis was conducted utilising 17 characters measured on the scolex and strobila of 101 specimens. Univariate statistics were first used to detect features that were useful for separating individual Eubothrium species and two different host populations of E. salvelini. Subsequent multivariate discriminant analysis, combining all the measured variables, made it possible to separate all four species. A comparison of the four taxa revealed that (1) E. fragile is the most distinct species, possessing a much smaller scolex than the other congeners, and its similarity with the other marine species E. crassum is not proven; (2) the two freshwater taxa, E. rugosum and E. salvelini are the most similar; (3) the characters most suitable for species differentiation are the length of the scolex, the width of the apical disc, the width of the neck and its area, the width of eggs and the number of testes; (4) the width of the apical disc was confirmed to be the most stable character at the intraspecific level (within E. salvelini host populations) and is therefore considered to be a trait of the highest discriminative power in the subset of four Eubothrium species

A molecular study of Eubothrium rugosum (Batsch, 1786) (Cestoda: Pseudophyllidea) using ITS rDNA sequences, with notes on the distribution and intraspecific sequence variation of Eubothrium crassum (Bloch, 1779)

The internal transcribed spacers (ITS-1 and ITS-2) of the ribosomal DNA (rDNA) of fish cestodes of the genus Eubothrium were sequenced. The ITS sequences of Eubothrium rugosum were determined and compared with previously analysed congeneric species, Eubothrium crassum and Eubothrium salvelini. The ITS-1 sequences of E. rugosum and E. crassum were 535 bp long, the length of E. salvelini ITS-1 was 536 bp. The ITS-2 region was found to be 403 bp in E. rugosum and E. crassum, and 401-402 bp in E. salvelini. The ITS-1 region of E. rugosum was closer to E. salvelini (identity 98.9-99.1%) than to E. crassum (97.9-98.1%), while the sequence similarity within the ITS-2 region was almost identical (97.5-98.0% for E. crassum; 97.7-98.3% for E. salvelini). Several restriction enzymes were found to be suitable for the differentiation of the three Eubothrium species by PCR-RFLP. The intraspecific sequence variation of E. crassum from different fish hosts (freshwater and marine) and European regions was very low, 0.2% for ITS-1 and 0.5% for ITS-2. Analysis of the ITS sequences of specimens from rainbow trout from three localities in Scotland revealed that both E. crassum and E. salvelini are present in this fish host

A comparative study of the egg morphology of four species of Eubothrium (Cestoda: Pseudophyllidea) with comments on their early development

Freshly released eggs from four species of the cestode Eubothrium (Eubothrium crassum, Eubothrium fragile, Eubothrium rugosum, and Eubothrium salvelini) were subjected to morphological and morphometric analysis. The eggs of the two freshwater species, E. rugosum and E. salvelini, were ovoid with a lobed embryophore whereas the eggs of the two marine species, E. crassum and E. fragile, were more circular with a smooth embryophore. However, the morphological differences between species were not readily evident to permit their clear distinction from one another. To discriminate species, a forward stepwise linear discriminant analysis, using six of the seven measured metric characters made on the eggs, was used, which gave 100% correct classification of two species, E. rugosum and E. salvelini, and a high proportion of correct classification for E. crassum (98%) and E. fragile (83%). Of the latter two species, one specimen of E. crassum and five specimens of E. fragile were misclassified between the respective groups. The principal characters used in the classification of the species were the width of the egg, the length of the mediolateral hooks, and the width of the oncosphere. To provide more information on the life cycle of each species, the eggs were used in a series of infection trials to identify appropriate intermediate hosts. Experimental infections with freshwater copepods were successful when exposed to the eggs of E. salvelini, partially successful when exposed to the eggs of marine E. crassum with 10% of the copepods becoming infected, but no infections were obtained when the eggs of E. fragile were used