Clinostomum complanatum (Rudolphi, 1814) is known as a food-borne zoonotic parasite using freshwater fish as intermediate host, although in its native distribution area in Europe, the Danube and Po basins and the northeast of Italy, no cases have been reported. This study reports a new geographic location outside the native distribution range of this parasite. We detected high infection levels of C. complanatum in wild perch (Perca fluviatilis Linnaeus) from the Doubs River in continental France. Clinostomum complanatum reached a prevalence of 97% and mean intensity of 5.8 cysts per fish in 33 fish examined, with most fish already infected by the age of two years. No correlations were observed between fish traits (standard length, weight, age and sex) and the number of parasites. The distribution of C. complanatum cysts in the fish body was not random. The tissue surrounding the opercula and branchial cavity, and muscles along the lateral line and near the pectoral, dorsal and caudal fins were the preferred sites of infection. Morphological identification was confirmed with molecular data. The French specimens shared identical mt cox1 haplotypes with specimens from Italy. The distribution of this exotic parasite in freshwaters in continental France deserves further research. Under the current French law, the parasite occurrence represents an economic loss for the local freshwater aquaculture industry, and a possible zoonotic risk for consumers if undetected, especially when raw or marinated fish is consumed. We provided a list of all reported hosts for C. complanatum from the western Palaearctic, discussed the knowledge gap regarding the zoonotic risk of C. complanatum in Europe, and highlighted the lack of policy that may increase the hazards posed by food-borne emerging parasites and their broader ecological impacts as alien species in freshwaters in Europe.

• Klíčová slova
• 28S rRNA gene, Halzoun syndrome, Platyhelminthes, alien species, cytochrome c oxidase subunit I mitochondrial gene, food-borne disease, freshwater ecosystems,
• MeSH
• infekce červy třídy Trematoda * veterinární   epidemiologie   parazitologie   MeSH
• nemoci ryb * parazitologie   epidemiologie   MeSH
• okounovití * parazitologie   MeSH
• řeky * MeSH
• Trematoda * izolace a purifikace   klasifikace   fyziologie   MeSH
• zoonózy parazitologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Francie epidemiologie   MeSH

Freshwater ecosystems in Lithuania are threatened by the introduction of invasive fish species including Neogobius fluviatilis, N. melanostomus, Perccottus glenii, and Pseudorasbora parva. Data on helminth parasites of these fishes have not been comprehensively studied, with only two reports on parasites of N. melanostomus from the Curonian Lagoon and Baltic Sea, Lithuania. We examined 278 fish individuals representing 4 invasive species from 13 waterbodies. Using morphological and molecular analyses, we identified 29 helminth taxa representing 15 digenean trematodes, 6 nematodes, 4 cestodes, 2 acanthocephalans, and 2 monogeneans. With 18 species, N. fluviatilis had the highest helminth diversity, followed by N. melanostomus (11 species) and Ps. parva (8 species). Perccottus glenii was found to be free from helminth infection. The availability of historical information on the native fish parasites in Lithuania allowed us to conclude that out of the 29 recorded species, invasive fish serve as hosts for 22 local fish helminth species, while 7 helminth species have been reported exclusively in invasive fish. Based on newly obtained and previously published data, a total of 34 helminth species parasitise invasive fish in Lithuania, of which 30 use these fish as intermediate or paratenic hosts.

• Klíčová slova
• Baltic freshwater ecosystems, Neogobius spp., Perccottus glenii, Pseudorasbora parva, aquatic invasion, morphological and molecular identification,
• Publikační typ
• časopisecké články MeSH

The genus Coitocaecum Nicoll, 1915 is part of the most speciose digenean family, the Opecoelidae Ozaki, 1925, which is found globally in both freshwater and marine fishes. Fifteen opecoelid species have been reported from marine fishes in South Africa, yet only one species of Coitocaecum has been described from this region: Coitocaecum capense Bray, 1987. During an explorative study of the digeneans of the endemic, intertidal fish Clinus superciliosus (Linnaeus) from the Saldanha Bay area, Cape Town harbour, Hermanus, the Tsitsikamma section of the Garden Route National Park and Chintsa East in South Africa, a total of three distinct species of Coitocaecum were identified based on morphological and molecular (28S rDNA, ITS1-5.8S-ITS2 rDNA and COI mtDNA) data: the previously mentioned C. capense, Coitocaecum brayi sp. n. and a third, unnamed species. We provide the first molecular characterisation of species of Coitocaecum from South Africa, accompanied by detailed morphological descriptions. This study illustrates the importance of an integrated taxonomic approach, especially when studying species with similar morphology. These findings further emphasise the lack of information on the true diversity and molecular data for trematodes of marine fishes in South Africa, creating a great capacity for future explorative taxonomic studies and highlighting the use of intertidal areas for conducting such research.

• Klíčová slova
• Afrotropical region, Trematoda, genetics, marine fish parasites, morphology, phylogeny,
• MeSH
• fylogeneze MeSH
• Perciformes * MeSH
• ribozomální DNA genetika   MeSH
• ryby MeSH
• Trematoda * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Jihoafrická republika MeSH
• Názvy látek
• ribozomální DNA MeSH

DNA sequence data became an integral part of species characterization and identification. Still, specimens associated with a particular DNA sequence must be identified by the use of traditional morphology-based analysis and correct linking of sequence and identification must be ensured. Only a small part of DNA sequences of the genus Diplostomum (Diplostomidae) is based on adult isolates which are essential for accurate identification. In this study, we provide species identification with an aid of morphological and molecular (cox1, ITS-5.8S-ITS2 and 28S) characterization of adults of Diplostomum baeri Dubois, 1937 from naturally infected Larus canus Linnaeus in Karelia, Russia. Furthermore, we reveal that the DNA sequences of our isolates of D. baeri are identical with those of the lineage Diplostomum sp. clade Q , while other sequences labelled as the ‘D. baeri’ complex do not represent lineages of D. baeri. Our new material of cercariae from Radix balthica (Linnaeus) in Ireland is also linked to Diplostomum sp. clade Q. We reveal that D. baeri is widely distributed in Europe; as first intermediate hosts lymnaeid snails (Radix auricularia (Linnaeus), R. balthica) are used; metacercariae occur in eye lens of cyprinid fishes. In light of the convoluted taxonomy of D. baeri and other Diplostomum spp., we extend the recommendations of Blasco-Costa et al. (2016, Systematic Parasitology 93, 295–306) for the ‘best practice’ in molecular approaches to trematode systematics. The current study is another step in elucidating the species spectrum of Diplostomum based on integrative taxonomy with well-described morphology of adults linked to sequences.

• Klíčová slova
• Ireland, Laridae, Lymnaeidae, Russia, nuclear and mitochondrial DNA, trematoda,
• MeSH
• Cyprinidae * MeSH
• fylogeneze MeSH
• infekce červy třídy Trematoda * parazitologie   veterinární   MeSH
• nemoci ryb * parazitologie   MeSH
• Trematoda * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH

A new species of Crassicutis Manter, 1936 (Digenea: Megaperidae) is described from the intestine of Satanoperca jurupari (Cichlidae) in the Amazon River basin, Brazil. The genus Crassicutis currently contains eight species. Crassicutis manteri n. sp. is morphologically very similar to Crassicutis cichlasomae Manter, 1936, a parasite of cichlids reported from Mexico, the Antilles, and Central and South America. Molecular data revealed that C. cichlasomae represents a species complex in Middle American cichlids. The new species can be readily distinguished from C. cichlasomae sensu lato, and the other congeners, by a combination of morphological traits such as a narrow, elongate mouth opening (versus spherical in other species), the tandem position of testes (symmetrical or oblique in most congeners), narrow body widening towards its posterior end (versus widely oval, leaf-like in other species), and short intestinal caeca ending close to the posterior end of the posterior testis (versus reaching more posteriorly in other species). Six novel sequences of 28S rDNA, ITS1, and cox1 were generated for two isolates of the new species. Sequences of the 28S rRNA gene were used to corroborate that Crassicutis is sister taxa of Homalometron Stafford, 1904. Mitochondrial DNA corroborated the distinction of the new species with previously sequenced congeners in Middle American cichlids; the interspecific divergence between the new species and the genetic lineages of C. cichlasomae was very high, varying between 23.7 and 27.2%. Biogeographical implications of our findings are briefly discussed including questionable validity of records of C. cichlasomae from South America.

• Klíčová slova
• Amazon River basin, Cichlidae, DNA, Megaperidae, Morphology, Phylogenetic analysis,
• MeSH
• cichlidy parazitologie   MeSH
• DNA helmintů genetika   MeSH
• druhová specificita MeSH
• fylogeneze MeSH
• infekce červy třídy Trematoda parazitologie   MeSH
• mitochondriální DNA genetika   MeSH
• nemoci ryb parazitologie   MeSH
• RNA ribozomální 28S genetika   MeSH
• Trematoda anatomie a histologie   klasifikace   genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Jižní Amerika MeSH
• Názvy látek
• DNA helmintů MeSH
• mitochondriální DNA MeSH
• RNA ribozomální 28S MeSH

BACKGROUND: Metacercariae of Diplostomum are important fish pathogens, but reliable data on their diversity in natural fish populations are virtually lacking. This study was conducted to explore the species diversity and host-parasite association patterns of Diplostomum spp. in a large riverine system in Europe, using molecular and morphological data. METHODS: Twenty-eight species of fish of nine families were sampled in the River Danube at Nyergesújfalu in Hungary in 2012 and Štúrovo in Slovakia in 2015. Isolates of Diplostomum spp. were characterised morphologically and molecularly. Partial sequences of the 'barcode' region of the cytochrome c oxidase subunit 1 (cox1) and complete sequences of the nicotinamide adenine dinucleotide dehydrogenase subunit 3 (nad3) mitochondrial genes were amplified for 76 and 30 isolates, respectively. The partial cox1 sequences were used for molecular identification of the isolates and an assessment of haplotype diversity and possible host-associated structuring of the most prevalent parasite species. New primers were designed for amplification of the mitochondrial nad3 gene. RESULTS: Only lens-infecting Diplostomum spp. were recovered in 16 fish species of five families. Barcoding of representative isolates provided molecular identification for three species/species-level genetic lineages, D. spathaceum, D. pseudospathaceum and 'D. mergi Lineage 2', and three single isolates potentially representing distinct species. Molecular data helped to elucidate partially the life-cycle of 'D. mergi Lineage 2'. Many of the haplotypes of D. spathaceum (16 in total), D. pseudospathaceum (15 in total) and 'D. mergi Lineage 2' (7 in total) were shared by a number of fish hosts and there was no indication of genetic structuring associated with the second intermediate host. The most frequent Diplostomum spp. exhibited a low host-specificity, predominantly infecting a wide range of cyprinid fishes, but also species of distant fish families such as the Acipenseridae, Lotidae, Percidae and Siluridae. The nad3 gene exhibited distinctly higher levels of interspecific divergence in comparison with the cox1 gene. CONCLUSIONS: This first exploration of the species diversity and host ranges of Diplostomum spp., in natural fish populations in the River Danube, provided novel molecular, morphological and host-use data which will advance further ecological studies on the distribution and host ranges of these important fish parasites in Europe. Our results also indicate that the nad3 gene is a good candidate marker for multi-gene approaches to systematic estimates within the genus.

• Klíčová slova
• Barcodes, Diplostomidae, Diplostomum, Europe, Freshwater fishes, Metacercariae, River Danube, cox1, nad3,
• MeSH
• fylogeneze MeSH
• genetická variace * MeSH
• mitochondriální DNA genetika   MeSH
• nemoci ryb parazitologie   MeSH
• oční čočka parazitologie   MeSH
• oční nemoci parazitologie   veterinární   MeSH
• řeky MeSH
• ryby parazitologie   MeSH
• taxonomické DNA čárové kódování * MeSH
• Trematoda genetika   izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Maďarsko epidemiologie   MeSH
• Slovenská republika epidemiologie   MeSH
• Názvy látek
• mitochondriální DNA MeSH

We report digeneans (Diplostomidae, Crassiphialinae) in the endangered freshwater fishes Valencia letourneuxi and Valencia robertae, endemics of Western Greece. Digenean metacercariae occurred in two forms in the abdominal cavity, excysted and encysted, the latter attached to the gonads, liver and alimentary tract. Parasites were, using morphological and molecular techniques, identified as two representatives of Crassiphialinae, specifically part of the Posthodiplostomum-Ornithodiplostomum clade. The spatial, seasonal, and age class variation in parasite prevalence was examined. Autumn parasite prevalence varied between the six populations sampled (18.2 to 100%). Seasonal prevalence at the two sites sampled quadannually peaked in autumn and reached its lowest value in spring; prevalence increased with size to 100% in young adult fish. We did not find a correlation between prevalence and host sex. Overall parasites' weight averaged 0.64% of the host's, while parasite weight increased with host weight. A comparison of relative condition and hepatosomatic and gonadosomatic indices of infected and metacercariae-free specimens showed that infection did not have a significant effect on host body condition and reproduction. Regarding the parasite's life cycle, planorbid gastropods are proposed as potential first intermediate hosts in view of the host's diet and occurrence data of molluscs in the ecosystem. This is the first record of a diplostomid digenean in valenciid fishes and of representatives of the Posthodiplostomum-Ornithodiplostomum clade in a native Greek freshwater fish. Our findings are discussed in conjunction to fish conservation interventions, since parasites may contribute to the decline of endangered species.

• Klíčová slova
• Digenea, Host body condition, Parasite prevalence, Planorbidae, Platyhelminthes, Valenciidae,
• MeSH
• Cyprinodontidae parazitologie   MeSH
• ekosystém MeSH
• infekce červy třídy Trematoda parazitologie   veterinární   MeSH
• měkkýši parazitologie   MeSH
• metacerkárie * MeSH
• nemoci ryb parazitologie   MeSH
• ohrožené druhy MeSH
• sladká voda MeSH
• stadia vývoje MeSH
• Trematoda klasifikace   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Řecko MeSH

Aporocotylid blood flukes conspecific with Aporocotyle margolisi Smith, 1967 were collected from the bulbus arteriosus of the North Pacific hake Merluccius productus (Ayres). This study revisits the morphology of A. margolisi, including drawings, measurements and scanning electron microscopy images, and provides for the first time molecular data for the large subunit of the ribosomal RNA (28S rDNA) and the mitochondrial cytochrome c oxidase subunit 1 (cox1) genes for this species. A 28S rDNA phylogenetic study of A. margolisi, and all available Aporocotyle spp., was also performed. The distribution range of A. margolisi is extended to the Pacific coast of the USA. We provide a morphological comparison of Aporocotyle spp. from the Pacific coast in North America as well as other Aporocotyle spp. infecting hake. Comparisons with the original description revealed that the new specimens of A. margolisi were considerably larger with respect to all morphological features, except for shorter spines. Molecular results showed a close relationship between A. margolisi and A. argentinensis Smith, 1969 from the Argentine hake Merluccius hubbsi Marini. The phylogenetic relationships of Aporocotyle spp. point to a possible co-speciation of hakes species and these blood fluke parasites.

• MeSH
• druhová specificita MeSH
• fylogeneze * MeSH
• Gadiformes parazitologie   MeSH
• respirační komplex IV genetika   MeSH
• RNA ribozomální 28S genetika   MeSH
• Trematoda anatomie a histologie   klasifikace   genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Oregon MeSH
• Tichý oceán MeSH
• Názvy látek
• respirační komplex IV MeSH
• RNA ribozomální 28S MeSH

Metacercariae of two species of Posthodiplostomum Dubois, 1936 (Digenea: Diplostomidae) were subjected to morphological and molecular studies: P. brevicaudatum (von Nordmann, 1832) from Gasterosteus aculeatus (L.) (Gasterosteiformes: Gasterosteidae), Bulgaria (morphology, cox1 and ITS1-5.8S-ITS2) and Perca fluviatilis L. (Perciformes: Percidae), Czech Republic (morphology, cox1, ITS1-5.8S-ITS2 and 28S); and P. centrarchi Hoffman, 1958 from Lepomis gibbosus (L.) (Perciformes: Centrarchidae), Bulgaria (morphology, cox1 and ITS1-5.8S-ITS2) and Slovakia (cox1 and ITS1-5.8S-ITS2). In addition, cercariae of P. cuticola (von Nordmann, 1832) from Planorbis planorbis (L.) (Mollusca: Planorbidae), Lithuania (morphology and cox1) and metacercariae of Ornithodiplostomum scardinii (Schulman in Dubinin, 1952) from Scardinius erythrophthalmus (L.) (Cypriniformes: Cyprinidae), Czech Republic, were examined (morphology, cox1, ITS1-5.8S-ITS2 and 28S). These represent the first molecular data for species of Posthodiplostomum and Ornithodiplostomum Dubois, 1936 from the Palaearctic. Phylogenetic analyses based on cox1 and ITS1-5.8S-ITS2, using O. scardinii as the outgroup and including the three newly-sequenced Posthodiplostomum spp. from Europe and eight published unidentified (presumably species-level) lineages of Posthodiplostomum from Canada confirmed the distinct status of the three European species (contrary to the generally accepted opinion that only P. brevicaudatum and P. cuticola occur in the Palaearctic). The subspecies Posthodiplostomum minimum centrarchi Hoffmann, 1958, originally described from North America, is elevated to the species level as Posthodiplostomum centrarchi Hoffman, 1958. The undescribed "Posthodiplostomum sp. 3" of Locke et al. (2010) from centrarchid fishes in Canada has identical sequences with the European isolates of P. centrarchi and is recognised as belonging to the same species. The latter parasite, occurring in the alien pumpkinseed sunfish Lepomis gibbosus in Europe, is also supposed to be alien for this continent. It is speculated that it colonised Europe long ago and is currently widespread (recorded in Bulgaria, Slovakia and Spain); based on the cox1 sequence of an adult digenean isolate from the Ebro Delta, Spain, only the grey heron (Ardea cinerea L.) (Ciconiiformes: Ardeidae) is known to be its definitive host in Europe.

• MeSH
• druhová specificita MeSH
• fylogeneze * MeSH
• Perciformes parazitologie   MeSH
• respirační komplex IV genetika   MeSH
• ribozomální DNA genetika   MeSH
• Smegmamorpha parazitologie   MeSH
• Trematoda anatomie a histologie   klasifikace   genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Evropa MeSH
• Názvy látek
• respirační komplex IV MeSH
• ribozomální DNA MeSH

BACKGROUND: The genus Diplostomum (Platyhelminthes: Trematoda: Diplostomidae) is a diverse group of freshwater parasites with complex life-cycles and global distribution. The larval stages are important pathogens causing eye fluke disease implicated in substantial impacts on natural fish populations and losses in aquaculture. However, the problematic species delimitation and difficulties in the identification of larval stages hamper the assessment of the distributional and host ranges of Diplostomum spp. and their transmission ecology. METHODS: Total genomic DNA was isolated from adult worms and shotgun sequenced using Illumina MiSeq technology. Mitochondrial (mt) genomes and nuclear ribosomal RNA (rRNA) operons were assembled using established bioinformatic tools and fully annotated. Mt protein-coding genes and nuclear rRNA genes were subjected to phylogenetic analysis by maximum likelihood and the resulting topologies compared. RESULTS: We characterised novel complete mt genomes and nuclear rRNA operons of two closely related species, Diplostomum spathaceum and D. pseudospathaceum. Comparative mt genome assessment revealed that the cox1 gene and its 'barcode' region used for molecular identification are the most conserved regions; instead, nad4 and nad5 genes were identified as most promising molecular diagnostic markers. Using the novel data, we provide the first genome wide estimation of the phylogenetic relationships of the order Diplostomida, one of the two fundamental lineages of the Digenea. Analyses of the mitogenomic data invariably recovered the Diplostomidae as a sister lineage of the order Plagiorchiida rather than as a basal lineage of the Diplostomida as inferred in rDNA phylogenies; this was concordant with the mt gene order of Diplostomum spp. exhibiting closer match to the conserved gene order of the Plagiorchiida. CONCLUSIONS: Complete sequences of the mt genome and rRNA operon of two species of Diplostomum provide a valuable resource for novel genetic markers for species delineation and large-scale molecular epidemiology and disease ecology studies based on the most accessible life-cycle stages of eye flukes.

• MeSH
• cestodózy epidemiologie   parazitologie   veterinární   MeSH
• fylogeneze MeSH
• genom mitochondriální * MeSH
• genom u helmintů * MeSH
• nemoci ryb epidemiologie   parazitologie   MeSH
• operon MeSH
• ploštěnci klasifikace   genetika   izolace a purifikace   MeSH
• RNA helmintů genetika   MeSH
• RNA ribozomální genetika   MeSH
• ryby MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• RNA helmintů MeSH
• RNA ribozomální MeSH