BACKGROUND: Monogenea (Platyhelminthes, Neodermata) are the most species-rich class within the Neodermata superclass of primarily fish parasites. Despite their economic and ecological importance, monogenean research tends to focus on their morphological, phylogenetic, and population characteristics, while comprehensive omics analyses aimed at describing functionally important molecules are few and far between. We present a molecular characterisation of monogenean representative Eudiplozoon nipponicum, an obligate haematophagous parasite infecting the gills of the common carp. We report its nuclear and mitochondrial genomes, present a functional annotation of protein molecules relevant to the molecular and biochemical aspect of physiological processes involved in interactions with the fish hosts, and re-examinate the taxonomic position of Eudiplozoon species within the Diplozoidae family. RESULTS: We have generated 50.81 Gbp of raw sequencing data (Illumina and Oxford Nanopore reads), bioinformatically processed, and de novo assembled them into a genome draft 0.94 Gbp long, consisting of 21,044 contigs (N50 = 87 kbp). The final assembly represents 57% of the estimated total genome size (~ 1.64 Gbp), whereby repetitive and low-complexity regions account for ~ 64% of the assembled length. In total, 36,626 predicted genes encode 33,031 proteins and homology-based annotation of protein-coding genes (PCGs) and proteins characterises 14,785 (44.76%) molecules. We have detected significant representation of functional proteins and known molecular functions. The numbers of peptidases and inhibitors (579 proteins), characterised GO terms (16,016 unique assigned GO terms), and identified KEGG Orthology (4,315 proteins) acting in 378 KEGG pathways demonstrate the variety of mechanisms by which the parasite interacts with hosts on a macromolecular level (immunomodulation, feeding, and development). Comparison between the newly assembled E. nipponicum mitochondrial genome (length of 17,038 bp) and other diplozoid monogeneans confirms the existence of two distinct Eudiplozoon species infecting different fish hosts: Cyprinus carpio and Carassius spp. CONCLUSIONS: Although the amount of sequencing data and characterised molecules of monogenean parasites has recently increased, a better insight into their molecular biology is needed. The E. nipponicum nuclear genome presented here, currently the largest described genome of any monogenean parasite, represents a milestone in the study of monogeneans and their molecules but further omics research is needed to understand these parasites' biological nature.

• Keywords
• Annotation, Assembly, Genome, Helminths, Host–parasite interaction, Illumina, Mitochondrial genome, Monogenea, Nanopore, Sequencing,
• MeSH
• Phylogeny MeSH
• Genomics MeSH
• Carps * genetics   MeSH
• Parasites * MeSH
• Trematoda * MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH

Gyrodactylid parasites were observed on non-native populations of North-American freshwater catfishes, Ameiurus nebulosus and Ameiurus melas (Siluriformes: Ictaluridae), at several sites in the Elbe River basin, Czech Republic, Europe. Using a combination of morphological and genetic analyses, the parasites infecting A. nebulosus were determined to be Gyrodactylus nebulosus, a North American parasite co-introduced to Europe along with its Ameiurus fish hosts. Subtle morphometrical differences, as well as seasonal variations, were observed among parasites collected from A. nebulosus and A. melas. The host-related variation was further supported through genetic analysis of the partial 18S rDNA, ITS1-5.8S-ITS2 and COI, showing 0.2, 3.0 and 4.8% divergence, respectively. Consistent genetic differences indicated there were two distinct genotypes. Subtle morphological differences associated with the shape of sickle toe, anchor root and ventral bar membrane, according to host species, also supported the description of a new cryptic species, Gyrodactylus melas n. sp., infecting A. melas. Multivariate morphometrical analysis of haptoral hard parts showed significant differences between the anchor lengths of G. nebulosus and G. melas n. sp. However, the measurements of the haptoral hard structures partially overlapped between species, limiting the usage of these parameters for species delineation.

• Keywords
• Bullhead catfish, Ictaluridae, North-America, gyrodactylids, non-native host,
• MeSH
• Biodiversity * MeSH
• Ictaluridae * MeSH
• Trematode Infections epidemiology   parasitology   veterinary   MeSH
• Fish Diseases epidemiology   parasitology   MeSH
• Prevalence MeSH
• Rivers MeSH
• Trematoda anatomy & histology   genetics   MeSH
• Introduced Species MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic epidemiology   MeSH

We describe a new species, Gyrodactylus ginestrae n. sp., a parasite of the big-scale sand smelt (Atherina boyeri) from the Black Sea. This is the third monogenean species known from this fish host, found at 70% prevalence, but at relatively low abundance (1.9), on fish gills and fins. The new species is, both morphologically and genetically, most similar to G. salinae, which parasitizes the killifish Aphanius fasciatus (Cyprinodontidae) in the Mediterranean region. These species differ in the size of the haptoral hard parts and the number of small spines of the male copulatory organ. For molecular characterization, the internal transcribed spacer 1 (ITS1), 5.8S rRNA gene, and the internal transcribed spacer 2 (ITS2) were sequenced, completed by a fragment of the COII gene, thereby representing the first molecularly characterized gyrodactylid species from the Black Sea. Phylogenetic reconstruction based on the ITS1-5.8S-ITS2 sequence data indicated the position of G. ginestrae n. sp. in the marine "rugiensis" group of G. (Paranephrotus) and G. (Neonephrotus) subgenera which is part of the monophyletic "long ITS1" group. Taking into account the similarity of G. ginestrae n. sp. to several monogeneans from the Atlantic and Mediterranean regions, we suggest the Boreal-Atlantic origin of this species.

• Keywords
• Brackish water, Gulf of Odessa, Gyrodactylidae, Molecular study, Phylogeny, Ponto-Caspian,
• MeSH
• DNA, Helminth genetics   MeSH
• Species Specificity MeSH
• Phylogeny * MeSH
• Osmeriformes parasitology   MeSH
• Animal Fins parasitology   MeSH
• DNA, Ribosomal genetics   MeSH
• Trematoda anatomy & histology   classification   genetics   MeSH
• Gills parasitology   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Black Sea MeSH
• Names of Substances
• DNA, Helminth MeSH
• DNA, Ribosomal MeSH

BACKGROUND: African tetras (Alestidae) belonging to Brycinus Valenciennes are known to be parasitized with monogeneans attributed to two genera, Annulotrema Paperna & Thurston, 1969 and Characidotrema Paperna & Thurston, 1968 (Dactylogyridae). During a survey of monogeneans parasitizing alestids, species of Characidotrema were collected in Cameroon, D. R. Congo, Senegal, South Africa, Sudan and Zimbabwe. This paper provides new morphological data and the first molecular analysis broadening our knowledge on the diversity of these parasites. RESULTS: Seven species (four known and three new) of Characidotrema are reported from two species of Brycinus: C. auritum n. sp. and C. vespertilio n. sp. from B. imberi (Peters); and C. brevipenis Paperna, 1969, C. nursei Ergens, 1973, C. pollex n. sp., C. spinivaginus (Paperna, 1973) and C. zelotes Kritsky, Kulo & Boeger, 1987 from B. nurse (Rüppell). Species identification was based on morphological analysis of the sclerotized structures supported by nuclear ribosomal DNA (partial 18S rDNA, ITS1, and 28S rDNA) sequence data. Morphological analysis confirmed that the most apparent character distinguishing species in the genus is the morphology of the male copulatory organ and vagina. Observations on the haptoral sclerotized elements of these parasites by means of phase contrast microscopy revealed the presence of a sheath-like structure relating to the ventral anchor, a feature that supplements the generic diagnosis of Characidotrema. Maximum Likelihood and Bayesian analyses of the large subunit (28S) rDNA sequences recovered Characidotrema species isolated from the two Brycinus hosts as monophyletic, and indicated a closer relationship of this group to monogeneans parasitizing African cyprinids (Dactylogyrus spp.) and cichlids (species of Cichlidogyrus Paperna, 1960, Scutogyrus Pariselle & Euzet, 1995, and Onchobdella Paperna, 1968) than to those from catfishes (species of Quadriacanthus Paperna, 1961, Schilbetrema Paperna & Thurston, 1968 and Synodontella Dossou & Euzet, 1993). The overall agreement between the morphological diversification of the MCOs and the molecular tree observed in this study indicates that significant phylogenetic signals for clarifying relationships among species of Characidotrema are present in the characteristics of the MCO. CONCLUSIONS: It seems that intra-host speciation is an important force shaping the present distribution and diversity of Characidotrema but further studies are necessary to confirm this hypothesis and assess questions related to the phylogeny of these parasites. To identify potential co-speciation events, co-phylogenetic analyses of these monogeneans and their alestid hosts are required.

• Keywords
• Africa, Alestidae, Brycinus, Characidotrema, DNA, Dactylogyridae, Diversity, Monogenea,
• MeSH
• Cichlids parasitology   MeSH
• Phylogeny * MeSH
• Host-Parasite Interactions * MeSH
• Lakes parasitology   MeSH
• Fish Diseases parasitology   MeSH
• DNA, Ribosomal genetics   MeSH
• Trematoda classification   isolation & purification   MeSH
• Gills parasitology   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Africa South of the Sahara MeSH
• Names of Substances
• DNA, Ribosomal MeSH

BACKGROUND: Monogenean flatworms are the main ectoparasites of fishes. Representatives of the species-rich families Gyrodactylidae and Dactylogyridae, especially those infecting cichlid fishes and clariid catfishes, are important parasites in African aquaculture, even more so due to the massive anthropogenic translocation of their hosts worldwide. Several questions on their evolution, such as the phylogenetic position of Macrogyrodactylus and the highly speciose Gyrodactylus, remain unresolved with available molecular markers. Also, diagnostics and population-level research would benefit from the development of higher-resolution genetic markers. We aim to offer genetic resources for work on African monogeneans by providing mitogenomic data of four species (two belonging to Gyrodactylidae, two to Dactylogyridae), and analysing their gene sequences and gene order from a phylogenetic perspective. RESULTS: Using Illumina technology, the first four mitochondrial genomes of African monogeneans were assembled and annotated for the cichlid parasites Gyrodactylus nyanzae, Cichlidogyrus halli, Cichlidogyrus mbirizei (near-complete mitogenome) and the catfish parasite Macrogyrodactylus karibae (near-complete mitogenome). Complete nuclear ribosomal operons were also retrieved, as molecular vouchers. The start codon TTG is new for Gyrodactylus and for Dactylogyridae, as is the incomplete stop codon TA for Dactylogyridae. Especially the nad2 gene is promising for primer development. Gene order was identical for protein-coding genes and differed between the African representatives of these families only in a tRNA gene transposition. A mitochondrial phylogeny based on an alignment of nearly 12,500 bp including 12 protein-coding and two ribosomal RNA genes confirms that the Neotropical oviparous Aglaiogyrodactylus forficulatus takes a sister group position with respect to the other gyrodactylids, instead of the supposedly 'primitive' African Macrogyrodactylus. Inclusion of the African Gyrodactylus nyanzae confirms the paraphyly of Gyrodactylus. The position of the African dactylogyrid Cichlidogyrus is unresolved, although gene order suggests it is closely related to marine ancyrocephalines. CONCLUSIONS: The amount of mitogenomic data available for gyrodactylids and dactylogyrids is increased by roughly one-third. Our study underscores the potential of mitochondrial genes and gene order in flatworm phylogenetics, and of next-generation sequencing for marker development for these non-model helminths for which few primers are available.

• Keywords
• Cichlidae, Cichlidogyrus, Clariidae, Gene order, Gyrodactylus, Macrogyrodactylus, Mitogenome, Monogenea, Monopisthocotylea, Phylogenomics,
• MeSH
• Cichlids parasitology   MeSH
• Phylogeny MeSH
• Genome, Mitochondrial MeSH
• Mitochondria classification   genetics   MeSH
• Platyhelminths genetics   MeSH
• Gene Order MeSH
• DNA, Protozoan chemistry   isolation & purification   metabolism   MeSH
• Protozoan Proteins classification   genetics   MeSH
• RNA, Ribosomal classification   genetics   MeSH
• Sequence Analysis, DNA MeSH
• High-Throughput Nucleotide Sequencing MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• DNA, Protozoan MeSH
• Protozoan Proteins MeSH
• RNA, Ribosomal 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.

• Keywords
• Digenea, Host body condition, Parasite prevalence, Planorbidae, Platyhelminthes, Valenciidae,
• MeSH
• Killifishes parasitology   MeSH
• Ecosystem MeSH
• Trematode Infections parasitology   veterinary   MeSH
• Mollusca parasitology   MeSH
• Metacercariae * MeSH
• Fish Diseases parasitology   MeSH
• Endangered Species MeSH
• Fresh Water MeSH
• Life Cycle Stages MeSH
• Trematoda classification   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Greece MeSH

Lake Tanganyika is well-known for its high species-richness and rapid radiation processes. Its assemblage of cichlid fishes recently gained momentum as a framework to study parasite ecology and evolution. It offers a rare chance to investigate the influence of a deepwater lifestyle in a freshwater fish-parasite system. Our study represents the first investigation of parasite intraspecific genetic structure related to host specificity in the lake. It focused on the monogenean flatworm Cichlidogyrus casuarinus infecting deepwater cichlids belonging to Bathybates and Hemibates. Morphological examination of C. casuarinus had previously suggested a broad host range, while the lake's other Cichlidogyrus species are usually host specific. However, ongoing speciation or cryptic diversity could not be excluded. To distinguish between these hypotheses, we analysed intraspecific diversity of C. casuarinus. Monogeneans from nearly all representatives of the host genera were examined using morphometrics, geomorphometrics and genetics. We confirmed the low host-specificity of C. casuarinus based on morphology and nuclear DNA. Yet, intraspecific variation of sclerotized structures was observed. Nevertheless, the highly variable mitochondrial DNA indicated recent population expansion, but no ongoing parasite speciation, confirming, for the first time in freshwater, reduced parasite host specificity in the deepwater realm, probably an adaptation to low host availability.

• MeSH
• Principal Component Analysis MeSH
• Bayes Theorem MeSH
• Biological Evolution MeSH
• Cichlids genetics   parasitology   MeSH
• Phylogeny MeSH
• Genetic Variation MeSH
• Haplotypes MeSH
• Host Specificity MeSH
• Lakes MeSH
• Parasites genetics   MeSH
• Platyhelminths genetics   MeSH
• Population Dynamics MeSH
• Gills parasitology   MeSH
• Geography MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Tanzania MeSH