Intraspecific variation in Gyrodactylus mediotorus and G. crysoleucas (Gyrodactylidae) from Nearctic shiners (Leuciscidae): evidence for ongoing speciation, host-switching, and parasite translocation.

A parasitological investigation of Cyprinella venusta and Notropis cf. stramineus sampled in Texas, USA, in the Guadalupe River, revealed the presence of Gyrodactylus crysoleucas Mizelle and Kritsky, 1967 on C. venusta, and Gyrodactylus mediotorus King, Marcogliese, Forest, McLaughlin & Bentzen, 2013 on both fish species. This represents new leuscicid fish hosts and locality records for these two gyrodactylids. Gyrodactylus crysoleucas previously identified from both non-native Californian Notemigonus crysoleucas and from farmed stocks in Minnesota demonstrated intraspecific variability in terms of morphology and genetics as a local adaptation associated with isolation by distance. Results further confirmed G. crysoleucas as alien in the western USA and suggested host-switching involving C. venusta and N. crysoleucas. Conservative morphology and genetics on the part of G. mediotorus from C. venusta and N. cf. stramineus (Guadalupe River) was observed, while higher genetic divergence in the ITS sequences associated with morphological discrepancy was found between the studied G. mediotorus specimens and those of Notropis hudsonius than when considering the parasites of Notropis texanus. The separation of G. mediotorus into geographical subgroups may indicate ongoing speciation linked to the Pleistocene glaciations in North America, and to hydrographic barriers that facilitated separate evolutionary paths leading to speciation. We suggest that deep investigations of Gyrodactylus populations will help to understand the speciation of these parasites and their adaptation to Nearctic fish hosts.

Title: Variation intraspécifique chez Gyrodactylus mediotorus et G. crysoleucas (Gyrodactylidae), parasites de ménés néarctiques (Leuciscidae) : preuves d'une spéciation en cours, d'un changement d'hôte et d'une translocation de parasites. Abstract: Une enquête parasitologique sur Cyprinella venusta et Notropis cf. stramineus échantillonnés au Texas, États-Unis, dans la rivière Guadalupe, a révélé la présence de Gyrodactylus crysoleucas Mizelle et Kritsky, 1967 sur C. venusta, et de Gyrodactylus mediotorus King, Marcogliese, Forest, McLaughlin & Bentzen, 2013 sur les deux espèces de poissons. Ceci représente de nouveaux poissons Leuciscidae hôtes et des nouvelles localités pour ces deux Gyrodactylidae. Gyrodactylus crysoleucas, identifié précédemment à partir de Notemigonus crysoleucas californiens non indigènes et de stocks d'élevage du Minnesota a démontré une variabilité intraspécifique en termes de morphologie et de génétique en tant qu'adaptation locale associée à l'isolement par la distance. Les résultats ont en outre confirmé que G. crysoleucas était exotique dans l'ouest des États-Unis et ont suggéré un changement d'hôte impliquant C. venusta et N. crysoleucas. Une morphologie et une génétique conservatrices pour G. mediotorus de C. venusta et N. cf. stramineus (rivière Guadalupe) ont été observées, tandis qu'une divergence génétique plus élevée dans les séquences ITS, associée à une divergence morphologique, a été trouvée chez les spécimens étudiés de G. mediotorus et de Notropis hudsonius lorsque l'on considère les parasites de Notropis texanus. La séparation de G. mediotorus en sous-groupes géographiques peut indiquer une spéciation continue liée aux glaciations du Pléistocène en Amérique du Nord et à des barrières hydrographiques qui ont facilité des chemins évolutifs séparés menant à la spéciation. Nous suggérons que des études approfondies sur les populations de Gyrodactylus aideront à comprendre la spéciation de ces parasites et leur adaptation aux poissons hôtes néarctiques.

Exploring the use of fatty acid ethyl esters as a potential natural solution for the treatment of fish parasitic diseases.

Alternatives to conventional chemical treatments for parasitic diseases in fish are needed. Microalgal-sourced fatty acid ethyl esters (FAEEs) have shown an antiparasitic effect against Gyrodactylus turnbulli infection in guppies. Here, we tested a range of commercial FAEEs of various carbon chain lengths and unsaturation levels against two fish parasites. Guppies and barramundi infected with G. turnbulli and Trichodina sp., respectively, were used. The most effective FAEE, after excluding those toxic to fish, was ethyl laurate (12:0). For both parasites, the LD50 was 18.75 µM within 250 min of incubation. Ethyl eicosapentaenoate (20:5n3) was the next most effective FAEE against G. turnbulli, and dihomo-γ-linolenic acid ethyl ester (20:3n6) and ethyl α-linolenate (18:3n3) were the next most effective against Trichodina sp. In addition, FAEEs prepared from the microalga Phaeodactylum tricornutum residue, after fucoxanthin extraction, were examined against Trichodina sp. infection in barramundi for the first time. LD85 and LD100 was achieved at 2.5 and 5 µL mL-1 of the FAEE preparation, respectively. In vivo, immersion of infected barramundi in 1.25 µL mL-1 of this preparation for 24 h reduced infection prevalence from 100% to 53% and was non-toxic to fish.

Parasites (Monogenea) of tilapias Oreochromis niloticus and Coptodon rendalli (Cichlidae) in a river spring in Brazil.

In the present study, we examined 30 individuals of introduced African cichlids, Oreochromis niloticus and Coptodon rendalli, collected in a river spring of the Pardo River, Paranapanema River basin, southeastern Brazil. Based on morphological and molecular analyses of the partial LSU rDNA gene, we identified four species of monogeneans, Cichlidogyrus tilapiae, C. thurstonae, C. mbirizei, and Scutogyrus longicornis on the gills of O. niloticus, whereas individuals of C. rendalli were infested only with C. papernastrema. This is the first record of C. mbirizei and C. papernastrema in tilapias from Brazil. The ecological consequences of the introduction of exotic species of tilapia such as O. niloticus and C. rendalli along with their monogenean parasites in a wild environment represented by a river spring are discussed. Our new molecular data on Cichlidogyrus and Scutogyrus contribute to the investigation of the phylogenetic interrelationships of these widely distributed genera of monogeneans since their species composition is still unsettled.

Title: Parasites (Monogenea) des tilapias Oreochromis niloticus et Coptodon rendalli (Cichlidae) dans une source au Brésil. Abstract: Dans la présente étude, nous avons examiné 30 individus de cichlidés africains introduits, Oreochromis niloticus et Coptodon rendalli, collectés dans une source fluviale du fleuve Pardo, bassin du fleuve Paranapanema, dans le sud-est du Brésil. Sur la base d'analyses morphologiques et moléculaires du gène partiel de l'ADNr LSU, nous avons identifié quatre espèces de monogènes, Cichlidogyrus tilapiae, C. thurstonae, C. mbirizei et Scutogyrus longicornis sur les branchies d'O. niloticus, alors que les individus de C. rendalli étaient infestés uniquement par C. papernastrema. Il s'agit du premier signalement de C. mbirizei et C. papernastrema chez des tilapias du Brésil. Les conséquences écologiques de l'introduction d'espèces exotiques de tilapia telles que O. niloticus et C. rendalli ainsi que leurs monogènes parasites dans un environnement sauvage représenté par une source fluviale sont discutées. Nos nouvelles données moléculaires sur Cichlidogyrus et Scutogyrus contribuent à l'étude des interrelations phylogénétiques de ces genres de monogènes largement distribués puisque leur composition spécifique est encore incertaine.

An inside out journey: biogenesis, ultrastructure and proteomic characterisation of the ectoparasitic flatworm Sparicotyle chrysophrii extracellular vesicles.

BACKGROUND: Helminth extracellular vesicles (EVs) are known to have a three-way communication function among parasitic helminths, their host and the host-associated microbiota. They are considered biological containers that may carry virulence factors, being therefore appealing as therapeutic and prophylactic target candidates. This study aims to describe and characterise EVs secreted by Sparicotyle chrysophrii (Polyopisthocotyla: Microcotylidae), a blood-feeding gill parasite of gilthead seabream (Sparus aurata), causing significant economic losses in Mediterranean aquaculture. METHODS: To identify proteins involved in extracellular vesicle biogenesis, genomic datasets from S. chrysophrii were mined in silico using known protein sequences from Clonorchis spp., Echinococcus spp., Fasciola spp., Fasciolopsis spp., Opisthorchis spp., Paragonimus spp. and Schistosoma spp. The location and ultrastructure of EVs were visualised by transmission electron microscopy after fixing adult S. chrysophrii specimens by high-pressure freezing and freeze substitution. EVs were isolated and purified from adult S. chrysophrii (n = 200) using a newly developed ultracentrifugation-size-exclusion chromatography protocol for Polyopisthocotyla, and EVs were characterised via nanoparticle tracking analysis and tandem mass spectrometry. RESULTS: Fifty-nine proteins involved in EV biogenesis were identified in S. chrysophrii, and EVs compatible with ectosomes were observed in the syncytial layer of the haptoral region lining the clamps. The isolated and purified nanoparticles had a mean size of 251.8 nm and yielded 1.71 × 108 particles · mL-1. The protein composition analysis identified proteins related to peptide hydrolases, GTPases, EF-hand domain proteins, aerobic energy metabolism, anticoagulant/lipid-binding, haem detoxification, iron transport, EV biogenesis-related, vesicle-trafficking and other cytoskeletal-related proteins. Several identified proteins, such as leucyl and alanyl aminopeptidases, calpain, ferritin, dynein light chain, 14-3-3, heat shock protein 70, annexin, tubulin, glutathione S-transferase, superoxide dismutase, enolase and fructose-bisphosphate aldolase, have already been proposed as target candidates for therapeutic or prophylactic purposes. CONCLUSIONS: We have unambiguously demonstrated for the first time to our knowledge the secretion of EVs by an ectoparasitic flatworm, inferring their biogenesis machinery at a genomic and transcriptomic level, and by identifying their location and protein composition. The identification of multiple therapeutic targets among EVs' protein repertoire provides opportunities for target-based drug discovery and vaccine development for the first time in Polyopisthocotyla (sensu Monogenea), and in a fish-ectoparasite model.

A new species of Urocleidoides Mizelle & Price, 1964 from the gills of Schizodon nasutus Kner, 1858 (Characiformes, Anostomidae) in southeastern Brazil.

Based on an integrative approach, this study describes a new species of Urocleidoides infesting Schizodon nasutus in the Paranapanema River basin, Brazil. The new species can be distinguished from its congeners by specific morphological features, including the shape of the male copulatory organ and accessory piece, the ventral bar shape, and the shape and size of the hooks. Molecular analyses, particularly of the 28S rDNA gene, suggest a close relationship between the new species and Urocleidoides paradoxus. The phylogenetic and taxonomic arrangement of Urocleidoides is discussed, as the analyses of the 28S rDNA and COI mtDNA resolved the genus as non-monophyletic, with Diaphorocleidus, Rhinoxenus, and Cacatuocotyle nested within it. Additional morphological and molecular data of other congeneric species are required to investigate the phylogenetic position and classification of Urocleidoides. This study underscores the significance of using integrative approaches in understanding host-parasite associations and phylogenetic relationships, contributing to the description of the freshwater fish parasite biodiversity in South America, particularly in the Paranapanema river basin.

Species of Dactylogyridae (Platyhelminthes: Monogenoidea) infecting marine fishes of Moreton Bay, Queensland, Australia, with proposals of Pleuronectitrema n. gen. and Ecnomotrema n. gen. and descriptions of 13 new species.

Twenty-nine dactylogyrid species were reported from teleosts collected during a survey of the parasites of fishes of Moreton Bay, Queensland, Australia, in 2016. Two new genera, Ecnomotrema n. gen. and Pleuronectitrema n. gen., were proposed, and 13 new species were illustrated and described: Atherinicus difficilis n. sp., Diversohamulus similis n. sp., and Ecnomotrema cetiosum n. sp. from Atherinomorus vaigiensis; Haliotrema apiculum n. sp. from Pempheris schwenkii; Haliotrema tugulduriforme n. sp. from Scarus ghobban; Lethrinitrema australiense n. sp., and Lethrinitrema lituus n. sp. from Lethrinus nebulosus; Tetrancistrum siganioides n. sp. from Siganus fuscescens; Ligophorus bostrychus n. sp. from Planiliza subviridis; Neohaliotrema gemmula n. sp. from Abudefduf vaigiensis; Neohaliotrema moretonense n. sp. from Ab. vaigiensis and Ab. bengalensis; and Pleuronectitrema spirula n. sp. from Pseudorhombus arsius (all from Moreton Bay); Pleuronectitrema kuwaitense Kritsky & Sey n. sp. was described from specimens collected from P. arsius in Kuwait during 1996. Ten new host records were recorded: Chauhanellus duriensis Lim, 1994 and Hamatopeduncularia thalassini Bychowsky & Nagibina, 1969 from Pararius proximus; Diplectanotrema sp. 1 and sp. 2 from Sillago maculata and Goniistius vestitus, respectively; Diversohamulus tricuspidatus Bychowsky & Nagibina, 1969 from At. vaigiensis; Hal. cf. dempsteri (Mizelle & Price, 1964) Young, 1968 from Prionurus microlepidotus; Hal. spirale Yamaguti, 1968 from Upeneus tragula; Ligophorus kaohsianghsieni (Gussev, 1962) Gussev, 1985 from Planiliza subviridis; and Neohaliotrema malayense Lim & Gibson, 2010 from Ab. bengalensis and Ab. whitleyi. Twenty-five new faunal records for Moreton Bay were recorded, including the new species listed above and C. duriensis, Diplectanotrema sp. 1 and sp. 2, Diversohamulus tricuspidatus Bychowsky & Nagibina, 1969, Glyphidohaptor phractophallus Kritsky, Galli, & Yang, 2007, Hal. cf. dempsteri, Hal. johnstoni Bychowsky & Nagibina, 1970, Hal. spirale, Yamaguti, 1968 Hamatopeduncularia thalassini Bychowsky & Nagibina, 1969, Lethrinitrema nebulosum Sun, Li, & Yang, 2014, Ligophorus kaohsianghsieni (Gussev, 1962) Gussev, 1985, Ligophorus parvicopulatrix Soo & Lim, 2012, and Neohaliotrema malayense Lim & Gibson, 2010. Three new combinations were proposed: Hal. spariense Roubal, 1981 as Euryhaliotrema spariense (Roubal, 1981) n. comb.; and Hal. arsiosa Venkatanarasaiah, 1984 and Hal. youngi Venkatanarasaiah, 1984 as Pleuronectitrema arsiosa (Venkatanarasaiah, 1984) n. comb. and Pleuronectitrema youngi (Venkatanarasaiah, 1984) n. comb., respectively; Haliotrema sp. of Zhang is transferred to Pleuronectitrema as Pleuronectitrema sp. Hal. ctenochaeti Yamaguti, 1968 was replaced with Hal. asymphylum n. nom. to remove it from homonymy with Hal. ctenochaeti Young, 1968. Pseudohaliotrematoides zancli Yamaguti, 1968 was transferred to Haliotrema as Haliotrema hawaiiense n. nom. Hal. zancli Yamaguti, 1968 was considered a junior subjective synonym of Hal. dempsteri.

Title: Espèces de Dactylogyridae (Platyhelminthes, Monogenoidea) infectant les poissons marins de Moreton Bay, Queensland, Australie, avec la proposition de Pleuronectitrema n. gen. et Ecnomotrema n. gen. et la description de 13 nouvelles espèces. Abstract: Vingt-neuf espèces de Dactylogyridae sont signalées chez des téléostéens collectés lors d'une étude des parasites des poissons de Moreton Bay, Queensland, Australie, en 2016. Deux nouveaux genres, Ecnomotrema n. gen. et Pleuronectitrema n. gen., sont proposés, et 13 nouvelles espèces sont illustrées et décrites : Atherinicus difficilis n. sp., Diversohamulus similis n. sp., et Ecnomotrema cetiosum n. sp. d'Atherinomorus vaigiensis ; Haliotrema apiculum n. sp. de Pempheris schwenkii ; Haliotrema tugulduriforme n. sp. de Scarus ghobban ; Lethrinitrema australiense n. sp. et Lethrinitrema lituus n. sp. de Lethrinus nebulosus, Tetrancistrum siganioides n. sp. de Siganus fuscescens ; Ligophorus bostrychus n. sp. de Planiliza subviridis ; Neohaliotrema gemmula n. sp. d'Abudefduf vaigiensis ; Neohaliotrema moretonense n. sp. d'Ab. vaigiensis et Ab. bengalensis ; et Pleuronectitrema spirula n. sp. de Pseudorhombus arsius (tous de Moreton Bay). Pleuronectitrema hawaiiense Kritsky & Sey n. sp. est décrit à partir de spécimens collectés de P. arsius au Koweït en 1996. Dix nouvelles combinaisons parasites-hôtes sont répertoriées : Chauhanellus duriensis Lim, 1994 et Hamatopeduncularia thalassini Bychowsky & Nagibina, 1969 de Pararius proximus ; Diplectanotrema sp. 1 et sp. 2, respectivement de Sillago maculata et Goniistius vestitus ; Diversohamulus tricuspidatus Bychowsky & Nagibina, 1969 d'At. vaigiensis ; Hal. cf. dempsteri (Mizelle & Price, 1964) Young, 1968 de Prionurus microlepidotus ; Hal. spirale Yamaguti, 1968 d'Upeneus tragula ; Ligophorus kaohsianghsieni (Gussev, 1962) Gussev, 1985 de Planiliza subviridis ; et Neohaliotrema malayense Lim & Gibson, 2010 d'Ab. bengalensis et Ab. whitleyi. Vingt-cinq nouveaux signalements fauniques pour la baie Moreton sont rapportés, y compris les nouvelles espèces énumérées ci-dessus et C. duriensis, Diplectanotrema sp. 1 et sp. 2, Diversohamulus tricuspidatus Bychowsky & Nagibina, 1969, Glyphidohaptor phractophallus Kritsky, Galli et Yang, 2007, Hal. cf. dempsteri, Hal. johnstoni Bychowsky et Nagibina, 1970, Hal. spirale, Hamatopeduncularia thalassini Bychowsky & Nagibina, 1969, Lethrinitrema nebulosum Sun, Li, & Yang, 2014, Ligophorus kaohsianghsieni (Gussev, 1962) Gussev, 1985, Ligophorus parvicopulatrix Soo & Lim, 2012, et Neohaliotrema malayense Lim et Gibson, 2010. Trois nouvelles combinaisons sont proposées : Hal. spariense Roubal, 1981 comme Euryhaliotrema spariense (Roubal, 1981) n. comb. ; Hal. arsiosa Venkatanarasaiah, 1984 et Hal. youngi Venkatanarasaiah, 1984, respectivement comme Pleuronectitrema arsiosa (Venkatanarasaiah, 1984) n. comb. et Pleuronectitrema youngi (Venkatanarasaiah, 1984) n. comb. ; Haliotrema sp. de Zhang est transféré à Pleuronectitrema sous le nom de Pleuronectitrema sp. Hal. ctenochaeti Yamaguti, 1968 est remplacé par Hal. asymphylum n. nom. pour supprimer l'homonymie avec Hal. ctenochaeti Young, 1968. Pseudohaliotrematoides zancli Yamaguti, 1968 est transféré à Haliotrema sous le nom de Haliotrema hawaiiense n. nom. Hal. zancli Yamaguti, 1968 est considéré comme un synonyme subjectif junior de Hal. dempsteri.

Morphological and molecular characterization of Encotyllabe vallei Monticelli, 1907 (Monopisthocotylea, Monogenea) from the gilthead seabream Sparus aurata Linnaeus (Teleostei, Sparidae) from the southwestern Mediterranean and notes on host specificity of the genus Encotyllabe Diesing, 1850.

Incomplete original descriptions, the unavailability or poor conditions of specimens and the lack of detailed redescriptions have caused the validity of several species of the genus Encotyllabe Diesing, 1850 to be questioned. To date, seven of the recognized species were described upon one or two specimens, hindering study of intraspecific variations. This was made worse by considering few morphoanatomical differences sufficient to erect new species. Among Encotyllabe spp. occurring in Mediterranean waters, E. vallei was first described from the gilt-head bream Sparus aurata (Sparidae) off Italy. Although beautifully illustrated for a paper from that century, morphometric data for E. vallei from the type-host S. aurata remain unavailable. Previous records of E. vallei provided either morphometrical or molecular data, and its validity was questioned. We provide a redescription of E. vallei based on newly collected specimens from the S. aurata from the southwestern Mediterranean (off Algeria) using integrative taxonomy. Analysis of cox1 sequences of E. vallei from S. aurata, compared to sequences from other sparid hosts, mainly Pagellus bogaraveo, revealed a divergence not exceeding 2%, suggesting a stenoxenic specificity for this monogenean. Given that P. bogaraveo is the type-host for Encotyllabe pagelli, we were tempted to suggest a synonymy between E. vallei and E. pagelli. We refrained from doing so because E. pagelli was first described from the Atlantic coast off Brest, France. Morphological data for Encotyllabe from P. bogaraveo are warranted assessing the host specificity of E. vallei and whether there might be a species complex within individual sparid fish species.

Six species of nonmazocraeid monogeneans including two new species from clupeiform fishes of Visakhapatnam coast, Bay of Bengal.

During a survey on monogenean parasites of clupeiform fishes of Visakhapatnam coast, Bay of Bengal, apart from typical mazocraeids, 6 species of nonmazocraeids, including two new species were encountered. These belonged to 5 genera distributed over 4 families: Family Gastrocotylidae: Engraulicola thrissocles from Thryssa mystax, T. setirostris, T. malabarica; Engraulicola longisomum n.sp. from Thryssa mystax, T. setirostris, T. malabarica; Pellonicola elongatus from Ilisha filigera; family Megamicrocotylidae: Megamicrocotyle chirocentrus from Chirocentrus dorab; family Microcotylidae: Microcotyle clupei n.sp. from Thryssa mystax and T. setirostris; family Diclidophoridae: Keralina opisthopterus from Opisthopterus tardoore. Engraulicola longisomum n. sp. differs from E. thrissocles in the larger body size, in the relatively smaller size of haptor and also in the number of clamps and alignment of reproductive organs. Microcotyle clupei n.sp. the first species of the genus to be reported from clupeiform fishes differs from other species of the genus, mainly in the presence of a circlet of spines on the oral sucker. Descriptions are provided for all the species collected. Megamicrocotyle johnstoni is synonymized with M. chirocentrus.

New species of Gyrodactylus von Nordmann, 1832 (Monogenoidea: Gyrodactylidae) from Gymnodiptychus dybowskii (Kessler, 1874) (Schizothoracinae) in the Kunes River (Yili River basin), China.

Yili River system hosts a diverse fauna of fishes and parasites. Gymnodiptychus dybowskii is a rare and endangered aboriginal cold-water fish inhabit in the Yili river system. Our research identified a new species Gyrodactylus gymnodiptychi n. sp. isolated from G. dybowskii in the Kunes River (Yili River, China). Morphological comparison revealed identifiable differences between the new species and other parasites, including Gyrodactylus aksuensis, and Gyrodactylus tokobaevi, which are two known parasites living in G. dybowskii inhabit in the Aksu River west of Frunze (Kyrgyzstan), as well as Gyrodactylus montanus living in Shizothorax intermedius inhabited in the Tadzhikistan or Uzbekistan. Especially, the dorsal bar of G. gymnodiptychi n. sp. was raised at both ends with a hollow, and its hamulus roots were curved inward. The BLASTN search of GenBank did not detect any other ITS1-5.8S-ITS2 rDNA sequences same as G. gymnodiptychi's. Using the Bayesian Information and Maximum Likelihood methods to analyze the ITS1-5.8S-ITS2 rDNA gene sequences, we constructed phylogenetic trees for G. gymnodiptychi n. sp. Accordingly, our morphological and molecular research indicated that G. gymnodiptychi n. sp. was not only a new species of parasites but also the first Gyrodactylus member identified in the Yili River in China.

Microcotyle justinei n. sp. (Monogenea: Microcotylidae) from the Gills of the Cardinal Fish Apogon imberbis (Teleostei: Apogonidae) off the Algerian Coast of the Western Mediterranean.

PURPOSE: A new monogenean Microcotyle justinei n. sp. (Monogenea: Microcotylidae) is described based on specimens found on the gill filaments of the cardinal fish Apogon imberbis (Apogonidae) off the Algerian coast of the Western Mediterranean. METHODS: Monogeneans were examined, measured and drawn for a comparative morphological study with other species of Microcotyle and characterised molecularly using a partial fragment of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The identification of fish was confirmed by molecular barcoding using the cox1 gene. RESULTS: The new species is distinguished from all other species of the genus by a combination of features, such as the number and size of the clamps, the shape and size of the genital atrium and the number of testes. The molecular analysis of the cox1 gene sequences showed that interspecific differences between Microcotyle justinei n. sp. and published sequences of Microcotyle spp. was greater than 8.8%, strongly suggesting that the new species is distinct from other congeners with sequences available on GenBank. CONCLUSION: The morphological and molecular analyses support the status of M. justinei as a new species. The present finding extends the list of Microcotyle spp. to 72.

The evolution of endoparasitism and complex life cycles in parasitic platyhelminths.

Within flatworms, the vast majority of parasitism is innate to Neodermata, the most derived and diversified group of the phylum Platyhelminthes.1,2 The four major lineages of Neodermata maintain various combinations of life strategies.3 They include both externally (ecto-) and internally feeding (endo-) parasites. Some lineages complete their life cycles directly by infecting a single host, whereas others succeed only through serial infections of multiple hosts of various vertebrate and invertebrate groups. Food sources and modes of digestion add further combinatorial layers to the often incompletely understood mosaic of neodermatan life histories. Their evolutionary trajectories have remained molecularly unresolved because of conflicting evolutionary inferences and a lack of genomic data.4 Here, we generated transcriptomes for nine early branching neodermatan representatives and performed detailed phylogenomic analyses to address these critical gaps. Polyopisthocotylea, mostly hematophagous ectoparasites, form a group with the mostly hematophagous but endoparasitic trematodes (Trematoda), rather than sharing a common ancestor with Monopisthocotylea, ectoparasitic epithelial feeders. Phylogenetic placement of the highly specialized endoparasitic Cestoda alters depending on the model. Regardless of this uncertainty, this study brings an unconventional perspective on the evolution of platyhelminth parasitism, rejecting a common origin for the endoparasitic lifestyle intrinsic to cestodes and trematodes. Instead, our data indicate that complex life cycles and invasion of vertebrates' gut lumen, the hallmark features of these parasites, evolved independently within Neodermata. We propose the demise of the traditionally recognized class Monogenea and the promotion of its two subclasses to the class level as Monopisthocotyla new class and Polyopisthocotyla new class.

l-fucoside localization in the gills of the genus Takifugu and its possible implication in the parasitism of Heterobothrium okamotoi (Monogenea: Diclidophoridae).

BACKGROUND: The monogenean parasite Heterobothrium okamotoi only parasitizes the gills of Takifugu rubripes. In this study, we hypothesized that the carbohydrates contribute to high host specificity of H. okamotoi. METHODS: T. rubripes, T. niphobles, T. snyderi, and T. pardalis were used for UEA I staining of the gills and an in vivo challenge test against H. okamotoi. To examine the effect of l-fucose, an in vitro detachment test was conducted using the host's gills. Additionally, fucosylated proteins were isolated from the membrane proteins of T. niphobles gills. RESULTS: The location of l-fucoside and the infection dynamics in four species were correlated to some extent; H. okamotoi detached relatively quickly from T. niphobles possessing l-fucoside both on the surface of the gills and in certain types of cells, including mucus cells, but detached slowly from T. snyderi possessing l-fucoside in only certain types of cells, including mucus cells. Under the conditions examined, H. okamotoi exhibited minimal detachment from T. rubripes and T. pardalis, and l-fucoside was not detected. The significantly higher detachment rate of H. okamotoi from the host's gills incubated in l-fucose-containing medium compared with the controls suggests that l-fucose in the non-host gills induced detachment of H. okamotoi. Four fucosylated proteins, including mucin5AC-like, were identified as potential factors for the detachment of H. okamotoi. CONCLUSIONS: Fucosylated proteins covering the surface of non-host gills might contribute to H. okamotoi detachment. GENERAL SIGNIFICANCE: This research shows the possible involvement of oligosaccharides in the host specificity of monogenean parasites.

Transcriptome Profile Analyses of Head Kidney in Roach (Rutilus rutilus), Common Bream (Abramis brama) and Their Hybrids: Does Infection by Monogenean Parasites in Freshwater Fish Reveal Differences in Fish Vigour among Parental Species and Their Hybrids?

Hybrid generations usually face either a heterosis advantage or a breakdown, that can be expressed by the level of parasite infection in hybrid hosts. Hybrids are less infected by parasites than parental species (especially F1 generations) or more infected than parental species (especially post-F1 generations). We performed the experiment with blood-feeding gill parasite Paradiplozoon homoion (Monogenea) infecting leuciscid species, Abramis brama and Rutilus rutilus, their F1 generation and two backcross generations. Backcross generations tended to be more parasitized than parental lines and the F1 generation. The number of differentially expressed genes (DEGs) was lower in F1 hybrids and higher in backcross hybrids when compared to each of the parental lines. The main groups of DEGs were shared among lines; however, A. brama and R. rutilus differed in some of the top gene ontology (GO) terms. DEG analyses revealed the role of heme binding and erythrocyte differentiation after infection by blood-feeding P. homoion. Two backcross generations shared some of the top GO terms, representing mostly downregulated genes associated with P. homoion infection. KEGG analysis revealed the importance of disease-associated pathways; the majority of them were shared by two backcross generations. Our study revealed the most pronounced DEGs associated with blood-feeding monogeneans in backcross hybrids, potentially (but not exclusively) explainable by hybrid breakdown. The lower DEGs reported in F1 hybrids being less parasitized than backcross hybrids is in line with the hybrid advantage.

Revealing the hidden diversity of Gyrodactylus communities (Monogenea, Gyrodactylidae) from Nearctic Catostomidae and Leuciscidae fish hosts (Teleostei, Cypriniformes), with descriptions of ten new species.

Despite the high diversity of freshwater fishes in the Nearctic region, little is known about the composition of their parasite communities. We addressed the diversity of viviparous monogeneans of Gyrodactylus parasitizing highly diversified cypriniform fish inhabiting Nearctic watersheds. Nowadays, a thorough assessment of Gyrodactylus spp. diversity requires both morphological traits and genetic data. A combination of taxonomically important haptoral features and sequences of the ITS regions and 18S rDNA revealed 25 Gyrodactylus spp. parasitizing two catostomid and 15 leuciscid species sampled in six distinct localities in the United States and Canada. These include ten Gyrodactylus species recognized as new to science and described herein (G. ellae n. sp., G. hamdii n. sp., G. hanseni n. sp., G. huyseae n. sp., G. kuchtai n. sp., G. lummei n. sp., G. mendeli n. sp., G. prikrylovae n. sp., G. scholzi n. sp., and G. steineri n. sp.), seven already known species, and finally eight undescribed species. Overall, Nearctic Gyrodactylus spp. exhibited haptoral morphotypes known from fish hosts worldwide and those apparently restricted to Nearctic Gyrodactylus lineages like the typical ventral bar with a median knob and a plate-like membrane, or the additional filament attached to the handles of marginal hooks. The integrative approach further evidenced possible ongoing gene flow, host-switching in generalist Gyrodactylus spp., and regional translocation of monogenean fauna through fish introductions. The study highlights the hitherto underexplored morphological and genetic diversity of viviparous monogeneans throughout the Nearctic region.

Title: Révélation de la diversité cachée des communautés de Gyrodactylus (Monogenea, Gyrodactylidae) des poissons hôtes néarctiques Catostomidae et Leuciscidae (Teleostei, Cypriniformes), avec la description de dix nouvelles espèces. Abstract: Malgré la grande diversité des poissons d'eau douce dans la région néarctique, on sait peu de choses sur la composition de leurs communautés de parasites. Nous avons abordé la diversité des monogènes vivipares du genre Gyrodactylus parasitant des poissons cypriniformes très diversifiés habitant les bassins versants néarctiques. De nos jours, une évaluation approfondie de la diversité de Gyrodactylus spp. nécessite à la fois des traits morphologiques et des données génétiques. Une combinaison de caractéristiques haptorales et de séquences taxonomiquement importantes des régions ITS et de l'ADNr 18S a révélé 25 espèces de Gyrodactylus sur deux espèces de catostomidés et 15 espèces de leuciscidés échantillonnées dans six localités distinctes aux États-Unis et au Canada. Celles-ci comprennent dix espèces de Gyrodactylus reconnues comme nouvelles pour la science et décrites dans cette étude (G. ellae n. sp., G. hamdii n. sp., G. hanseni n. sp., G. huyseae n. sp., G. kuchtai n. sp., G. lummei n. sp., G. mendeli n. sp., G. prikrylovae n. sp., G. scholzi n. sp. et G. steineri n. sp.), sept espèces déjà connues et enfin huit espèces non décrites. Dans l'ensemble, les espèces de Gyrodactylus néarctiques présentaient des morphotypes haptoraux connus chez les poissons hôtes du monde entier et ceux apparemment limités aux lignées néarctiques de Gyrodactylus comme la barre ventrale typique avec un bouton médian et une membrane en forme de plaque, ou le filament supplémentaire attaché aux poignées des crochets marginaux. L'approche intégrative a en outre mis en évidence le flux de gènes en cours, le changement d'hôte chez les espèces de Gyrodactylus généralistes et la translocation régionale de la faune monogène par l'introduction de poissons. L'étude met en évidence la diversité morphologique et génétique jusqu'ici sous-explorée des monogènes vivipares dans toute la région néarctique.

Dactylogyrus spp. (Dactylogyridae, Monogenea) from tinfoil barb, Barbonymus schwanenfeldii imported into South Africa: morphometric and molecular characterisation.

This study reports on three species of Dactylogyrus Diesing, 1850 (Dactylogyridae) collected from tinfoil barb, Barbonymus schwanenfeldii (Bleeker) which were imported into South Africa as ornamental fish from Sri Lanka and Thailand. Supplementary morphometric characterisation and molecular data (partial 18S and 28S rDNA, and ITS1 region sequences) are presented for Dactylogyrus lampam (Lim & Furtado, 1986), Dactylogyrus tapienensis Chinabut & Lim, 1993 and Dactylogyrus viticulus Chinabut & Lim, 1993. Prevalence of Dactylogyrus spp. infection was 87% and 80% for fish from Sri Lanka and Thailand, respectively. Composition of the parasites between the fish of each origin differed. All three species were found to infect fish from Thailand, but only D. lampam was present on the fish received from Sri Lanka. Phylogenetic analysis revealed the position of studied species, with D. lampam clustering within the lineages of varicorhini-type species, while D. tapienensis and D. viticulus form a sister lineage to Dactylogyrus spp. associated with Cyprinus carpio L. and Carassius spp., species parasitising central African large cyprinids (Labeo Cuvier), and species parasitising African and Middle Eastern Carasobarbus spp.

Title: Dactylogyrus spp. (Dactylogyridae, Monogenea) de Barbonymus schwanenfeldii importé en Afrique du Sud : caractérisation morphométrique et moléculaire. Abstract: Cette étude porte sur trois espèces de Dactylogyrus Diesing, 1850 (Dactylogyridae), prélevées sur des Barbonymus schwanenfeldii (Bleeker) qui ont été importés en Afrique du Sud comme poissons d'ornement depuis le Sri Lanka et la Thaïlande. Une caractérisation morphométrique et des données moléculaires supplémentaires (ADNr 18S et 28S partiels et séquences de la région ITS1) sont présentées pour Dactylogyrus lampam (Lim & Furtado, 1986), Dactylogyrus tapienensis Chinabut & Lim, 1993 et Dactylogyrus viticulus Chinabut & Lim, 1993. La prévalence de l'infection par les Dactylogyrus spp. était respectivement de 87 % et 80 % pour les poissons du Sri Lanka et de Thaïlande. La composition des parasites entre les poissons des deux origines différait. Les trois espèces infectaient les poissons de Thaïlande, mais seul D. lampam était présent sur les poissons du Sri Lanka. L'analyse phylogénétique a révélé la position des espèces étudiées, D. lampam se regroupant dans les lignées d'espèces de type varicorhini, tandis que D. tapienensis et D. viticulus forment une lignée sœur des Dactylogyrus spp. associés à Cyprinus carpio L. et Carassius spp., espèces parasitant les grands cyprinidés d'Afrique centrale (Labeo Cuvier), et espèces parasitant les Carasobarbus spp. d'Afrique et du Moyen-Orient.

Nearctic Dactylogyrus species (Platyhelminthes, Monogenea) parasitizing cypriniform fishes in the context of morphology and phylogeny, with descriptions of seven new species.

Ribosomal DNA sequences are currently available for 32 morphologically recognized species of Dactylogyrus parasitizing Nearctic cypriniforms, but only 16 of them are assigned to nominal species. Herein, morphological data on 28 of the 32 species are provided, together with comments on their phylogenetic relationships in the context of the morphology of taxonomically important structures. Seven new species of Dactylogyrus are described from five species of leuciscids and one species of catostomids, as follows: D. aduncus n. sp. from Campostoma spadiceum, D. cloutmani n. sp. from Luxilus chrysocephalus isolepis, D. cornifrons n. sp. from Cyprinella venusta, D. fimbratus n. sp. from Rhinichthys cataractae, D. mcallisteri n. sp. from Semotilus atromaculatus, and D. chieni n. sp. and D. haneki n. sp. from Hypentelium nigricans. Four previously described species of Dactylogyrus, D. atromaculatus from S. atromaculatus, D. eos from C. neogaeus, D. parvicirrus from Notemigonus crysoleucas and D. perlus from L. c. isolepis, are redescribed and/or figured. As for the remaining 17 species, only the male copulatory organs (MCOs) taken from the respective hologenophores are illustrated. On the basis of phylogenetic analyses, two main clades of Nearctic Dactylogyrus were recognized and supported by the different morphology of the MCO. The first one included 22 strictly Nearctic species sharing the same MCO type with Dactylogyrus spp. parasitizing cyprinids likely of Asian origin. The second clade encompassed Dactylogyrus spp. with diverse MCO morphology and was placed in the sister position to Dactylogyrus spp. parasitizing European leuciscids and North-West African cyprinids.

Title: Espèce néarctique de Dactylogyrus (Platyhelminthes, Monogenea) parasitant des poissons cypriniformes dans un contexte de morphologie et phylogénie, avec description de sept nouvelles espèces. Abstract: Des séquences d'ADN ribosomal sont actuellement disponibles pour 32 espèces morphologiquement reconnues de Dactylogyrus parasitant les cypriniformes néarctiques, mais seulement 16 d'entre elles sont attribuées à des espèces nominales. Ici, des données morphologiques sur 28 des 32 espèces sont fournies avec des commentaires sur leurs relations phylogénétiques dans le contexte de la morphologie des structures taxonomiquement importantes. Sept nouvelles espèces de Dactylogyrus sont décrites à partir de cinq espèces de leuciscidés et d'une espèce de catostomidés, comme suit : D. aduncus n. sp. de Campostoma spadiceum, D. cloutmani n. sp. de Luxilus chrysocephalus isolepis, D. cornifrons n. sp. de Cyprinella venusta, D. fimbratus n. sp. de Rhinichthys cataractae, D. mcallisteri n. sp. de Semotilus atromaculatus et D. chieni n. sp. et D. haneki n. sp. de Hypentelium nigricans. Quatre espèces de Dactylogyrus précédemment décrites, D. atromaculatus de S. atromaculatus, D. eos de C. neogaeus, D. parvicirrus de Notemigonus crysoleucas et D. perlus de L. c. isolepis, sont redécrites et/ou figurées. Comme pour les 17 espèces restantes, seuls les organes copulateurs mâles (OCM) prélevés sur les hologénophores respectifs sont illustrés. Sur la base d'analyses phylogénétiques, deux clades principaux de Dactylogyrus néarctiques ont été reconnus et soutenus par des morphologies différentes de l'OCM. Le premier comprenait 22 espèces strictement néarctiques partageant le même type d'OCM que les Dactylogyrus spp. parasitant des cyprinidés probablement d'origine asiatique. Le deuxième clade comprenait des Dactylogyrus spp. avec diverses morphologies d'OCM et était placé en position sœur des Dactylogyrus spp. parasitant les leuciscidés européens et les cyprinidés d'Afrique du Nord-Ouest.

Amended diagnosis, mitochondrial genome, and phylogenetic position of Sphyranura euryceae (Neodermata, Monogenea, Polystomatidae), a parasite of the Oklahoma salamander.

Polystomatidae is a monogenean family whose representatives infect mainly (semi)-aquatic tetrapods. Species of Sphyranura Wright, 1879 exhibit ectoparasitism on salamander hosts, with molecular work supporting their inclusion within Polystomatidae, at an early diverging, yet unresolved, position in the clade of otherwise endoparasitic polystomatid parasites of batrachian hosts. Records of representatives of Sphyranura are scarce with genetic data only available for S. oligorchis Alvey, 1933. Based on detailed morphological examination and comparison with type material, we identified worms belonging to Sphyranura infecting Oklahoma salamander (Eurycea tynerensis) as S. euryceae Hughes & Moore, 1943. Along with an amended diagnosis of Sphyranura, we provide the first molecular data for S. euryceae in the form of a mitochondrial genome and nuclear (18S, 28S rRNA) markers. Close morphological similarity between the two species of Sphyranura is reflected in low genetic divergence. Mitochondrial level comparison reveals instances of tRNA gene rearrangements in polystomatids. Although the phylogenetic reconstruction supports Sphyranura as early branching in the lineage of polystomatid monogeneans infecting batrachians, certain nodes remain unresolved.

Title: Diagnostic modifié, génome mitochondrial et position phylogénétique de Sphyranura euryceae (Neodermata, Monogenea, Polystomatidae), un parasite de la salamandre de l'Oklahoma. Abstract: Les Polystomatidae sont une famille de monogènes dont les représentants infectent principalement les tétrapodes (semi)-aquatiques. Les espèces de Sphyranura Wright, 1879 présentent un ectoparasitisme sur les hôtes salamandres, et des travaux moléculaires soutiennent leur inclusion dans les Polystomatidae, à une position divergente précoce mais non résolue dans le clade des Polystomatidae endoparasites d'hôtes batraciens. Les signalements des représentants de Sphyranura sont rares et les données génétiques ne sont disponibles que pour S. oligorchis Alvey, 1933. Sur la base d'un examen morphologique détaillé et d'une comparaison avec le matériel type, nous avons identifié les vers appartenant à Sphyranura infectant la salamandre de l'Oklahoma (Eurycea tynerensis) comme S. euryceae Hughes & Moore, 1943. Parallèlement à un diagnostic modifié de Sphyranura, nous fournissons les premières données moléculaires pour S. euryceae sous la forme d'un génome mitochondrial et de marqueurs nucléaires (ARNr 18S, 28S). La similitude morphologique étroite entre les deux espèces de Sphyranura se traduit par une faible divergence génétique. La comparaison au niveau mitochondrial révèle des cas de réarrangements des gènes des ARNt chez les Polystomatidae. Bien que la reconstruction phylogénétique soutienne Sphyranura comme un rameau précoce dans la lignée des monogènes Polystomatidae infectant les batraciens, certains nœuds restent non résolus.

Sharing is caring? Barcoding suggests co-introduction of dactylogyrid monogeneans with Nile tilapia and transfer towards native tilapias in sub-Saharan Africa.

Invasive Nile tilapias negatively impact native tilapia species through hybridisation and competition. However, the co-introduction of parasites with Nile tilapia, and subsequent changes in parasite communities, are scarcely documented. Monogeneans are known pathogens of cultured Nile tilapia, although little is known about their fate once Nile tilapias establish in new ecosystems. We investigate the parasitological consequences of Nile tilapia introduction on native tilapias in basins in Cameroon, the Democratic Republic of the Congo (DRC), and Zimbabwe, focusing on ectoparasitic dactylogyrids (Monogenea). Using the mitochondrial cytochrome oxidase c subunit I (COI) and nuclear 18S-internal transcribed spacer 1 (18S-ITS1) rDNA region of 128 and 166 worms, respectively, we evaluated transmission of several dactylogyrid species. Parasite spillover from Nile tilapia was detected for Cichlidogyrus tilapiae to Coptodon guineensis in Cameroon, Cichlidogyrus thurstonae to Oreochromis macrochir in the DRC, and Cichlidogyrus halli and C. tilapiae to Coptodon rendalli in Zimbabwe. Parasite spillback to Nile tilapia was detected for Cichlidogyrus papernastrema and Scutogyrus gravivaginus from Tilapia sparrmanii and Cichlidogyrus dossoui from C. rendalli or T. sparrmanii in the DRC, and Cichlidogyrus chloeae from Oreochromis cf. mortimeri and S. gravivaginus from O. macrochir in Zimbabwe. 'Hidden' transmissions (i.e. transmission of certain parasite lineages of species that are naturally present on both alien and native hosts) were detected for C. tilapiae and Scutogyrus longicornis between Nile tilapia and Oreochromis aureus and C. tilapiae between Nile tilapia and Oreochromis mweruensis in the DRC, and Cichlidogyrus sclerosus and C. tilapiae between Nile tilapia and O. cf. mortimeri in Zimbabwe. A high density of Nile tilapia occurring together with native tilapias, and the broad host range and/or environmental tolerance of the transmitted parasites, are proposed as factors behind parasite transmission through ecological fitting. However, continuous monitoring and the inclusion of environmental variables are necessary to understand the long-term consequences of these transmissions on native tilapias and to elucidate other underlying factors influencing these transmissions.

An insight into the functional genomics and species classification of Eudiplozoon nipponicum (Monogenea, Diplozoidae), a haematophagous parasite of the common carp Cyprinus carpio.

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.

Monogenoidea Communities Differ Between Male Morphotypes of Bluegill Sunfish (Lepomis macrochirus).

PURPOSE: Bluegill sunfish (Lepomis macrochirus) are a North American sport fish that exhibit two different male morphotypes. Alpha-males are large, colorful, territorial, and have significant parental investment, while ß-males are small, drab, and have two reproductive phenotypes, neither of which involves parental investment. Due to the two different reproductive strategies, the level of interaction between congenerics varies, which may impact parasites that are spread via close contact, such as Monogenoidea that infect the gills. Monogeneans are ectoparasites on the gills and skin of fish hosts that may cause significant pathology in high numbers and be an indicator of host behavior and interaction between hosts. METHODS: In this study, 328 L. macrochirus (106 α-males, 92 ß-males, and 130 females) were necropsied from 8 lakes and ponds in northwestern Virginia to identify and enumerate monogenean parasites from the gills. RESULTS: Alpha-males had a significantly greater parasite abundance and species richness compared to ß-males. This may be due to the increased size and surface area of gills in α-males, increased interaction with females during mating, and stationary behavior when guarding nests, which enhanced the chances of α-males contracting the parasites. This also led to significant differences in the monogenean communities infecting the two morphotypes, which were also significantly influenced by the size of the hosts. CONCLUSIONS: It is important in future studies regarding parasitism that behavioral morphotypes within a sex, such as α-male and ß-male L. macrochirus in this study, be treated separately as behavioral and morphometric differences between them can potentially lead to differences in parasitism.