Petkevi ci ut_ e, R., Stun z_ enas, V., Stanevi ci ut_ e, G., Zhokhov, A.E. (2015). European Phyllodistomum (Digenea, Gorgoderidae) and phylogenetic affinities of Cercaria duplicata based on rDNA and karyotypes. 44,[191][192][193][194][195][196][197][198][199][200][201][202] Genetic markers of some European Gorgoderidae species, including potential adults of Cercaria duplicata, were obtained and used to clarify phylogenetic affinities within the genus Phyllodistomum and to verify conflicting data existing on their life cycles. Molecular data and karyotype, 2n = 18, provide further support for ascription of C. duplicata to the Gorgoderinae. Sequences of C. duplicata form a robustly supported major clade, phylogenetically distinct from other known gorgoderid species in both ITS2-and 28S-based phylogenetic trees. The molecular data revealed no match between C. duplicata and any species of Phyllodistomum, including adults found in the experimental studies. One of them, P. elongatum, showed no differences from type species P. folium. Other, P. angulatum, forms a robustly supported clade, which is closely related to P. macrocotyle clade in all phylogenetic trees. This study supports the concept that only P. macrocotyle is a parasite of Dreissena polymorpha among the phyllodistomes and life cycle described for the type species P. folium by Sinitsin (1905) can apparently be discounted. Previously reported low host specificity of P. folium was justified. Adults of P. folium were detected in eight teleost species from five families and four orders. Cystocercous cercariae of P. folium were recorded in sphaeriid bivalves of the genus Sphaerium and Pisidium. According to our molecular data, P. simile, parasite of bullhead, must be regarded as synonym of P. folium. Phyllodistomum umblae is most closely related to P. folium in the all phylogenetic analyses. Molecular phylogenies support a presumption that Phyllodistomum species with cystocercous cercariae developing in sphaeriid bivalves should be regarded as Phyllodistomum sensu stricto. The results reveal a clear need for reconsideration of the knowledge on gorgoderid life cycles based on experimental studies and re-evaluation of the validity of some nominal Phyllodistomum species. Corresponding author: Romualda Petkevi ci ut_ e,
BackgroundHost-specificity patterns are not well-defined for trematodes of the genus Phyllodistomum Braun, 1899. The Eurasian ruffe, Gymnocephalus cernuus L., has been recorded as a definitive host for Phyllodistomum folium (Olfers, 1816), P. angulatum Linstow, 1907 and P. megalorchis Nybelin, 1926 and as the type-host for P. pseudofolium Nybelin (1926). A wide range of other host fishes have been recorded for these species as well. All present host records have been based on light microscopy and the life-cycles of P. pseudofolium, P. angulatum and P. megalorchis are unknown. The validity of P. pseudofolium and P. megalorchis require verification. In this study, rDNA sequences generated from adult Phyllodistomum spp., as well as from larval stages developing in Pisidium amnicum Müller, were analysed to establish the real number of Phyllodistomum species utilizing G. cernuus, and to associate larvae with the corresponding adult forms.ResultsPhylogenetic analyses of adult and larval stages of Phyllodistomum spp. based on ITS2 and partial 28S rDNA data allowed the confirmation of the validity of P. pseudofolium. A macrocercous cercaria, known as Phyllodistomum sp. from P. amnicum is genetically identical to adult P. pseudofolium. Phyllodistomum megalorchis obtained from its type-host, Lota lota L., showed no genetic differences from P. angulatum parasitizing Sander lucioperca L. In our analysis, P. pseudofolium, P. angulatum and P. macrocotyle formed a highly supported clade despite the fact that these species appear to be associated with distinct patterns of first intermediate host identity and cercarial morphology. Some morphological differences between gravid specimens of P. pseudofolium and P. angulatum were observed and their SEM tegumental surface topography is described.ConclusionsThe results lead us to the perception that macroevolutionary host switching in the genus Phyllodistomum is independent of host phylogeny. This study suggests strict host-specificity (oioxeny) for P. pseudofolium using one first intermediate host species (P. amnicum) and one definitive host species (G. cernuus). Phyllodistomum megalorchis is to be regarded as a synonym of P. angulatum. The close phylogenetic relatives, P. pseudofolium and P. angulatum, can be differentiated by morphological traits, the micromorphology and tegumental surface topography of these two species is intended to provide useful data for their identification and support the use of such features as a valuable taxonomic criterion. Molecular data showed that G. cernuus is a definitive host for two species: the oioxenous P. pseudofolium and the euryxenous P. folium.
BackgroundWithin the genus Crepidostomum Braun, 1900, identification of species and taxonomic decisions made only on the basis of adult morphology have resulted in great problems associated with evaluating actual diversity and validity of species. Life-cycle data, while equal in importance to adult characters, are scarce, controversial or incomplete for most Crepidostomum spp. In this study, rDNA sequences generated from adult and larval Crepidostomum spp. and some other allocreadiid species were analysed to reveal the diversity and phylogenetic relationships of the species and their host range. Detailed morphological description based on light microscopy, SEM tegumental surface topography and genetic data are provided for the poorly known trematode C. oschmarini Zhokhov & Pugacheva, 1998 found in the intestine of two teleost fish species, Barbatula barbatula (L.) and Cottus gobio L.ResultsWe characterized 27 isolates of adult and larval parasites. Based on newly obtained 28S and ITS1-5.8S-ITS2 rDNA sequences, new intermediate and final hosts were ascertained, and life-cycles clarified for some allocreadiids. New knowledge on the diversity and phylogenetic relationships of European Crepidostomum spp. was gained. The validity of C. oschmarini was verified based on comparative sequence analysis. Ophthalmoxiphidiocercariae of C. oschmarini were recorded in sphaeriid bivalves Pisidium (Euglesa) casertanum (Poli). Additionally, morphological differences between gravid specimens of C. oschmarini and other related species were observed.ConclusionsSpecies of the Allocreadiidae parasitizing fishes in Europe are distributed among two monophyletic genera, Allocreadium and Bunodera, and two paraphyletic Crepidostomum clades. A complex of Crepidostomum metoecus (syn. C. nemachilus), C. oschmarini and Crepidostomum sp. 2 clustered in one clade, and a complex of C. farionis, Crepidostomum sp. 1 and, probably, C. wikgreni in the other. Molecular data indicated that C. oschmarini and Crepidostomum sp. 2 presumably have a wide geographical distribution in Europe. The new data provided evidence that Crepidostomum is a more diverse genus than can be judged from morphological data and host switching in this genus may occur independently of fish-host phylogeny.
In the present study, phylogenetic relationships of European and Far Eastern representatives of the genus Aspidogaster Baer, 1827 were analysed: A. conchicola Baer, 1827, A. limacoides Diesing, 1834, A. ijimai Kawamura, 1915 and A. chongqingensis Wei, Huang & Dai, 2001. Based on ITS1-5.8S-ITS2 rDNA sequence data, an obvious differentiation was seen between specimens of A. limacoides sensu stricto from the European part of Russia and A. limacoides sensu Chen et al., 2010 from China (13.7%); the latter parasites were recognized as A. chongqingensis. Aspidogaster chongqingensis was more closely related to A. ijimai than to A. limacoides s. str. Specimens of A. ijimai from the Amur River, Khanka Lake (Russian Far East) and China were grouped into a single clade with low intra specific molecular differentiation (d = 0-0.3%). Specimens of A. conchicola from the European part of Russia, the Russian Far East and China also formed a single distinct clade. Genetic differentiation between European and Chinese samples of this species was two times lower (d = 0.45%) than between Russian Far East and European or Chinese samples (d = 0.96%), suggesting a long-term separate existence of A. conchicola in the Russian Far East.
Spermatological characters of the digenean Brandesia turgida (Brandes, 1888), an intestinal parasite of the frog Pelophylax ridibundus (Pallas, 1771), have been investigated by means of transmission electron microscopy for the first time. The process of the spermatozoon formation begins with the appearance of the differentiation zone bordered by cortical microtubules and containing two centrioles associated with striated rootlets and with an intercentriolar body. The intercentriolar body is made up of seven distinct electron-dense plates, two less electron-dense, and four electron-lucent zones. The orthogonal development of the two flagella is followed by a flagellar rotation and their proximodistal fusion with the median cytoplasmic process. This process is accompanied by an extension of both the mitochondrion and nucleus into the median cytoplasmic process. The mature spermatozoon of B. turgida contains two parallel axonemes of unequal lengths with the 9 + "1" trepaxonematan pattern, mitochondrion, nucleus, parallel cortical microtubules, four electron-dense attachment zones, an external ornamentation of the plasma membrane, and electron-dense glycogen granules. The anterior extremity of the male gamete contains one complete centriole, a small component of the central element of the second centriole, and peripheral cortical microtubules (up to 45). The posterior extremity of the mature spematozoon exhibits tubular elements of the disorganized axoneme. The present study provides the first data on spermiogenesis within the family Pleurogenidae. Variations of the spermatozoa ultrastructural characters within Digenea, in particular, between different families of the superfamily Microphalloidea, are discussed.
Data on trematode life cycles, fish host distribution, transmission, and fish and snail infection are reported in this review. European freshwater trematodes of the genus Sanguinicola (Aporocotylidae) remain an insufficiently studied group of trematodes. Five species of Aporocotylidae (Sanguinicola armata, S. inermis, S. intermedia, S. volgensis, and S. rutili) in freshwater fish of Europe are described. In addition, they have been found in the water bodies of Central Asia and West Siberia (Ob-Irtysh River basin). The life cycle allowing us to assign the cercariae and adults to a certain species is known only for S. armata, S. inermis, and S. rutili. Trematodes of the genus Sanguinicola are found in 26 fish species assigned to 7 families and 4 orders and 24 gastropod species assigned to 7 families. With few exceptions, the sanguinicolid infection of fish and snails is rather low in the natural water bodies.
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