Morphometric comparison of three members of the Gyrodactylus nemachili-like species group (Monogenea: Gyrodactylidae) on Barbatula barbatula L. in the Czech Republic, with a reinstatement of G. papernai Ergens & Bychowsky, 1967

The Gyrodactylus nemachili-like species complex, parasitising stone loach Barbatula barbatula L. in the Czech Republic, was investigated based on the detailed morphometric analysis of its haptoral hard parts. The existence of two species, G. jiroveci Ergens & Bychowsky, 1967 and G. pseudonemachili Ergens & Bychowsky, 1967 was confirmed. A standard discriminant analysis of the morphometric data showed a clear separation between specimens of G. jiroveci and another species or variant. Based on the morphology of the haptoral hard parts, this third species was more similar to G. jiroveci than to G. pseudonemachili. The differences between G. jiroveci and the third species were mainly in the morphology of the marginal hook sickle, but included the total length of the marginal hook, the length of the hamulus point, the length of the marginal hook sickle, the width of the ventral bar, and the width of the dorsal bar. Based on comparison with the original material, the third species was identified as G. papernai Ergens & Bychowsky, 1967, which has recently been considered a synonym of G. jiroveci. These analyses provided sufficient evidence for the restoration of G. papernai as a valid taxon. Partial sequences of the internal transcribed spacer rRNA genes (ITS rDNA) verified the validity of all three Gyrodactylus species studied.     

First report of Gyrodactylus spp. (Platyhelminthes: Monogenea) in the western Mediterranean Sea: molecular and morphological descriptions

The Gyrodactylus spp. fauna on species of gobies, Pomatoschistus, Gobiusculus, and Knipowitschia (Gobiidae: Teleostei), from the western Mediterranean and Adriatic seas is strikingly similar to that found in the Baltic Sea and eastern Atlantic Ocean, both in morphology and in internal transcribed spacer (ITS) rRNA. The fauna consisted of Gyrodactylus branchialis, G. ostendicus, G. gondae, G. rugiensis, G. rugiensoides, and G. arcuatus. No new species have been found. A morphometric comparison between G. branchialis from the Mediterranean and Adriatic seas and its type locality in Ostend (Belgium) showed significant differences in ventral bar and marginal hook features. The morphometric variation was lower in G. rugiensis, whereas no significant differences were found in G. ostendicus. Gyrodactylus branchialis and G. ostendicus collected on P. microps were slightly different in the ITS rDNA (-0.6%) compared with specimens on the closely related P. marmoratus, probably reflecting ongoing speciation. A hybrid zone was identified in the Vaccarès lagoon complex (France) where both host species are sympatric. There was no clear geographic or host-related pattern in the variation found in the ITS2 rDNA in G. arcuatus sampled from P. microps, G. flavescens, Pungitius pungitius, K. panizzae, and its original host Gasterosteus aculeatus (2 polymorphic sites). Of all studied species, only G. arcuatus, G. rugiensis, and G. rugiensoides showed minor intraspecific variation in the ITS rDNA. Hence, the physical separation by a shoreline of more than 10,000 km is hardly reflected in the parasite ITS rDNA.     

Gyrodactylus medaka n. sp. (Monogenea: Gyrodactylidae) parasitic on wild and laboratory-reared medaka Oryzias latipes (Beloniformes: Adrianichthyidae) in Japan

Gyrodactylus medaka n. sp. (Monogenea: Gyrodactylidae) is described from the skin, fins, and gills of medaka Oryzias latipes (Beloniformes: Adrianichthyidae) from Japan. This new species was collected from wild medaka in Hiroshima, Aichi, Saga, and Kumamoto prefectures, and laboratory-reared medaka in Chiba and Aichi prefectures. The small marginal hook sickle (≤4?μm) and the length of the marginal hook of the new species are the diagnostic morphological characters differentiated from other gyrodactylids reported from Asia. The pairwise sequence divergences for the interspecific variation in ITS regions and the phylogenetic analysis suggest that the populations of G. medaka n. sp. may have a similar genetic variation as the medaka populations in Japan. Gyrodactylus medaka n. sp. and Dactylogyrus oryziasi (Monogenea: Dactylogyridae) can maintain their populations in laboratory aquaria using medaka as their hosts, and these monogeneans and medaka have the potential as experimental model animals for clarifying various aspects of their host-parasite relationships. In addition, we report the composition of modified ammonium picrate-glycerin (APG) and show it is advantageous for monogenean taxonomy.     

Morphology and molecular taxonomy of Gyrodactylus jennyae n. sp. (Monogenea) from tadpoles of captive Rana catesbeiana Shaw (Anura), with a review of the species of Gyrodactylus Nordmann, 1832 parasitising amphibians

Gyrodactylus jennyae n. sp. is described from the body surface and mouthparts of tadpoles of the bullfrog Rana catesbeiana Shaw imported presumably from Missouri, USA, into a federal government facility in Moncton, New Brunswick, Canada. Its morphology resembles most closely that of G. chologastris Mizelle, Whittaker & McDougal, 1969 described from two amblyopsids (blind cave fishes) in Kentucky and North Carolina. Both species have long slender hamuli, a ventral bar with a relatively long membrane and small anterolateral processes, a cirrus with two rows of small spines and marginal hooks with a well-developed sickle heel and short handle. The two species differ morphologically; G. jennyae has a marginal hook sickle with a more pronounced heel than that found in G. chologastris. A BLAST search using a 945 base pair sequence that included the nuclear ribosomal DNA internal transcribed spacers 1 and 2 and the 5.8S rRNA gene from G. jennyae n. sp. showed that the overall similarity with other Gyrodactylus sequences on GenBank was relatively low. The ITS1 region was similar to that of G. misgurni Ling, 1962; however, no ITS2 and 5.8S rRNA sequences are available for that species. A separate search using 5.8S sequences revealed that G. markakulensis Gvosdev, 1950 and G. laevis Malmberg, 1957 were the closest to G. jennyae (1 and 2 bp differences, respectively). These species are parasites of cyprinids (or their predators) and are similar to G. jennyae and G. chologastris in having a double row of small hooks on the cirrus and overall similar morphologies of the haptoral hard parts. There are now five species of Gyrodactylus described exclusively from amphibians and this appears to have involved at least three separate host-switches from fishes.     

Gyrodactylus sommervillae n. sp. (Monogenea) from Abramis brama (L.) and Rutilus rutilus (L.) (Cyprinidae) in Oxfordshire,UK

Gyrodactylus sommervillae n. sp. (Monogenea) is described from the skin, fins and gills of Abramis brama (L.) and Rutilus rutilus (L.) from Blenheim Palace Lake, Oxfordshire, UK. G. sommervillae n. sp. closely resembles G. aphyae Malmberg, 1957, G. kearni Ergens, 1990 and G. lamberti Ergens, 1990 in the approximate shape of the marginal hook sickle and the anchors, but can be separated on the precise shape of the marginal hooks and the ventral bar.     

Gyrodactylus neili n. sp. (Monogenea: Gyrodactylidae), a parasite of chain pickerel Esox niger Lesueur (Esocidae) from freshwaters of New Brunswick, Canada

Gyrodactylus neili n. sp. (Monogenea: Gyrodactylidae) is described from the fins and body surface of Esox niger Lesueur (chain pickerel) (Esocidae) from the St. Croix River drainage, New Brunswick, Canada. G.␣neili n. sp. resembles most closely G. fryi Cone & Dechtiar, 1984, a parasite of E. masquinongy in North America, in having relatively large thin hamuli, well-developed marginal hook sickles with a relatively long, wide blade and short handle, a ventral bar with small antero-lateral processes and tongue-shaped membrane, and a cirrus with many small spines in two rows. The new species is easily separated from G. fryi by the length of the hamuli (70-76 m versus 92 m, respectively), by the distal width of the sickle (7–9 versus 14-16 m, respectively) and by subtle differences in the shape of the toe and heel of the marginal hook sickle. Sequence data (922 bp) of rDNA (internal transcribed spacers 1 and 2 and 5.8S) of G. neili n. sp. returned no identical matches in GenBank. The 5.8 sequence alone, however, was identical to morphologically similar gyrodactylids of the subgenus Gyrodactylus from cyprinid fishes in Eurasia. The discovery of G. neili n. sp. and features of its genetic makeup support the idea that this lineage parasitised ancestral cyprinids and that it radiated, possibly through predator/prey interactions, to an ancestor of contemporary Esox. It is concluded that DNA comparison of monogeneans on Holarctic freshwater hosts, such as E.␣lucius, may shed light on the nature of speciation of these parasites.     

Gyrodactylus anarhichatis n. sp. and G. microanchoratus n. sp. (Monogenea: Gyrodactylidae) from the Atlantic wolf-fish Anarhichas lupus (Anarhichadidae) off the coast of Norway

The gyrodactylid monogeneans Gyrodactylus anarhichatis n. sp. and G. microanchoratus n. sp. are reported from the skin and fins of wild Atlantic wolf-fish Anarhichas lupus L. caught off the coast of northern Norway. G. anarhichatis n. sp. is also reported from farmed Atlantic wolf-fish and spotted wolf-fish A. minor Olafsen. G. anarhichatis n. sp. most closely resembles G. corti Mizelle & Kritsky, 1967, but is also similar to G. perlucidus Bychowsky & Poljansky, 1953 and G. errabundus Malmberg, 1970, while G. microanchoratus n. sp. differs from most other Gyrodactylus spp. by having marginal hooks longer than anchors and a peculiar lump on the marginal hook sickle toe pointing towards the sickle point.     

Gyrodactylus pictae n. sp. (Monogenea: Gyrodactylidae) from the Trinidadian swamp guppy Poecilia picta Regan, with a discussion on species of Gyrodactylus von Nordmann, 1832 and their poeciliid hosts

Gyrodactylus pictae n. sp. is recorded from Poecilia picta in heterospecific shoals with the guppy P.reticulata in Northern Trinidad. G. pictae is morphologically similar to G. turnbulli Harris, 1986, but the hamuli and marginal hooks are slightly smaller and more gracile. The toe and the point of the marginal hook have a distinctly different shape, providing the best morphological characters for distinguishing the two species. The rDNA ITS1 and ITS2 sequences differ from those of G. turnbulli (the closest relative) by >5, suggesting that these two taxa are not sibling species. The origin of the two species on poeciliids of the subgenus Micropoecilia is discussed, and it is suggested that this may represent a case of host–parasite co-evolution.     

The description of Gyrodactylus mulli sp. n. (Monogenea: Gyrodactylidae) from the Black Sea blunt-snouted mullet Mullus barbatus ponticus

A new species Gyrodactylus mulli sp. n. is described from the thorax fins of blunt-snouted mullet Mullus barbatus ponticus by the collections of B. E. Bychowsky made in 1947 near Karadag on the Black Sea. Gyrodactylus mulli sp. n. differs from G. alviga Dmitrieva et Gerasev, 2000 (described from blunt-snouted mullet too) in having another morphotype of the marginal hooks. The new species differs from G. proterorhini Ergens, 1967, which has the identical type of the marginal hooks, in having longer membrane of the ventral connective bar, longer point of the anchors, shorter marginal hooks, and lesser size of the cirrus.     

Two new species of Gyrodactylus von Nordmann, 1832 from Profundulus oaxacae (Pisces: Profundulidae) from Oaxaca,Mexico, studied by morphology and molecular analyses

In the present study, two new species of Gyrodactylus are described from Profundulus oaxacae, a fish endemic to the Pacific slope of Oaxaca State, Mexico. Fishes were collected within their distribution range in 5 localities in the Atoyac-Verde River. Gyrodactylus montealbani n. sp. and G. zapoteco n. sp. were erected and characterized morphologically (sclerites of the attachment apparatus and the male copulatory organ) and molecularly (sequences of the Internal Transcribed Spacer region of rDNA). The haptoral sclerites of the new species are similar to those of Gyrodactylus iunuri and Gyrodactylus tepari, both recently described from the goodeid fish Goodea atripinnis, from the Mexican States of Jalisco and Querétaro, respectively; and to Gyrodactylus xtachuna described from the poeciliid Poeciliopsis gracilis in Veracruz State, Mexico – nonetheless, these species can all be discriminated based on their marginal hook morphology. Specimens of G. montealbani n. sp. and G. zapoteco n. sp. were sequenced, and were aligned with sequences of 25 other Gyrodactylus spp. Both Maximum likelihood and Bayesian inference analyses indicated that the two new species are members of independent, well-supported lineages – these are the first Gyrodactylus species described from Profundulus oaxacae.     

Gyrodactylus quadratidigitus n. sp. (Monogenea: Gyrodactylidae), a parasite of the leopard-spotted goby Thorogobius ephippiatus (Lowe) from the south-western coast of the UK

Gyrodactylus quadratidigitus n. sp. (Monogenea, Gyrodactylidae) is described from the gills, fins and skin of Thorogobius ephippiatus (Lowe) (leopard-spotted goby) from Portland Castle Bay, Dorset, UK. G. quadratidigitus n. sp. most closely resembles the species ofGyrodactyluspreviously described by Geets from Pomatoschistus microps(Krøyer) in the approximate shape and size of the attachment hooks but can be readily distinguished from other species of the genus by the distinctive square shape of the toe region of the marginal hook sickle, the anteriorly positioned cirrus bulb and the unusually short intestinal crura which extend only as far as the level of the testes.     

Molecular markers for gyrodactylids (Gyrodactylidae: Monogenea) from five fish families (Teleostei)

Thirty-one gyrodactylid species from five families of freshwater fish were examined and variable region V4 of the 18S small subunit ribosomal RNA gene and ribosomal RNA internal transcribed spacers ITS1 and ITS2 were sequenced. Both the V4 region and spacers ITS1 and ITS2 proved useful for gyrodactylid diagnosis. Sequences of these fragments exhibited interspecific variations and allowed clear determination at the species level. In some cases, the length of the ITS1 PCR fragment provided useful genetic markers. Species that yielded a short ITS1 fragment also showed distinct groupings in ITS2 and V4 sequences that were markedly different to sequences from species that contain a long ITS1. Repetitive sequences located in the ITS1 of Gyrodactylus gobii and Gyrodactylus vimbi accounted for some of the variations in length of PCR products. There was no evidence for intraspecific variation within these regions and short tandem repeats were not found in the other species studied. The number of polymorphic and intraspecific variations in nucleic acid sequences was low, therefore these variations did not affect species determination of gyrodactylids. Minor differences in the sequences between Western and Eastern European populations were detected for Gyrodactylus salaris/Gyrodactylus thymalli, Gyrodactylus teuchis and Gyrodactylus truttae, but these do not affect species diagnosis based on ribosomal DNA sequence. These results confirm the utility of both variable region V4 and the ITS as molecular markers for Gyrodactylus species.     

Molecular and morphological comparisons between Gyrodactylus ostendicus n. sp. (Monogenea: Gyrodactylidae) on Pomatoschistus microps (Krøyer) and G. harengi Malmberg, 1957 on Clupea harengus membras L.

Gyrodactylus ostendicus n. sp. was exclusively found on fins of the common goby Pomatoschistus microps (Krøyer). The haptoral hard parts are among the smallest described for species of Gyrodactylus. A presumed similarity between the new species and G. harengi Malmberg, 1957 (subgenus Metanephrotus Malmberg, 1964) encouraged a comparative approach. A morphological analysis showed the marginal hook sickles of G. ostendicus to be of quite a different type and similar to those of G. arcuatus Bychowsky sensu Bychowsky &; Poljansky (1953) (subgenus Mesonephrotus Malmberg, 1964). The new species has a pharynx with short pharyngeal processes. Its protonephridial system has small bladders, indicating an association with the subgenera Mesonephrotus or Metanephrotus. Molecular phylogenetic analyses, including all of the species of Mesonephrotus and Metanephrotus currently available on the GenBank database, suggested that the new species belongs to Mesonephrotus. Combined morphological and molecular studies of the new species show that G. ostendicus is more closely related to G. arcuatus than to G. harengi.     

Description of Gyrodactylus banmae n. sp. (Monogenea,Gyrodactylidae) parasitic on zebrafish,Danio rerio

A new species of Gyrodactylus was described on the body surface of zebrafish (Danio rerio) in China. Basing on morphological characteristics and ITS sequence, we identified the parasite as a new member of the Gyrodactylus-wageneri group. Morphologically, Gyrodactylus sp. nov. is greatly similar to “G. zebrae”, another species parasitic on zebrafish: both have moderately stout hamulus, marginal hook sickle with a prominent heel and toe, as well as a curved blade. However, distinct haptoral shape differences were detected between these two species. The dorsal bar is straight in Gyrodactylus sp. nov. but strongly curved in “G. zebrae”, and the sickle shaft in Gyrodactylus sp. nov. is approximately perpendicular to the base, but in “G. zebrae” it is slanted downwards. The ITS1-5.8S-ITS2 sequence also indicate that Gyrodactylus sp. nov. exhibits the highest similarity to “G. zebrae”: 95.7% sequence identity suggests interspecific differentiation. Phylogenetic analysis of the ITS1-ITS2 sequence showed that Gyrodactylus sp. nov. formed a sister clade with “G. zebrae”, and exhibited a relatively close phylogenetic relationship with G. kobayashii, G. gurleyi, and G. longoacuminatus, all of which parasitise on goldfish, Carassius auratus. To test the susceptibility of zebrafish and goldfish to the Gyrodactylus sp. nov., ten gyrodactylids were inoculated to the caudal fin of zebrafish and goldfish. The gyrodactylids exhibited the ability to attach themselves to the goldfish, and some gyrodactylids reproduced a few days after the inoculation. On day 9, however, the mean abundance sharply decreased to zero on goldfish and increased to more than 30 on zebrafish. The result suggested that golfish is an unsuitable host for Gyrodactylus sp. nov. Therefore, on the basis of morphology, molecular sequence similarity, and host susceptibility, we conclude that the gyrodactylid found on the zebrafish is a new species, which we named Gyrodactylus banmae.     

A new gyrodactylid species from Cobitis granoei (Rendahl) (Cobitidae) in central China

Gyrodactylus granoei n. sp. (Monogenea: Gyrodactylidae) is described from the fins and body surface of the spine loach, Cobitis granoei (Rendahl) (Cobitidae), in central China. It resembles a suite of species from cyprinid and cobitid fishes that have short, compact hamuli, a ventral bar with no obvious anterolateral projections, a linguiform posterior membrane, a male copulatory organ with small hooks in multiple rows, a simple cylindrical dorsal bar, and short marginal hooks, with an expanded sickle heel. Of these, the new species resembles most closely Gyrodactylus micracanthus Hukuda, 1940 from Misgurnus anguillicaudatus (Cantor) (Cobitidae), but it is identified by the length of the hamuli and the morphology of dorsal and ventral bars. When searched using BLAST, sequence data (829 bp) spanning the ITS1, 5.8s, and ITS2 region did not return an identical match; close similarity (82-88%) was found with sequenced members of the subgenus Gyrodactylus. It is suggested that the new species is part of a freshwater lineage that has radiated successfully among cyprinid fishes in North America, Europe, and Asia, and some of their predator and amphibian neighbors.     

Redescription of Gyrodactylus teuchis Lautraite,Blanc, Thiery,Daniel & Vigneulle, 1999 (Monogenea: Gyrodactylidae); a species identified by ribosomal RNA sequence

Molecular and morphological features of Gyrodactylus specimens from Oncorhynchus mykiss, Salmo trutta and Salmo salar were examined. Sequences from variable region V4 of the small subunit ribosomal RNA gene and the ribosomal RNA internal transcribed spacers, produced by the FRS Marine Laboratory, revealed that these were not the same as other well-characterised Gyrodactylus that are common on European salmonids and were in fact a distinct species. Initial morphological examination of the opisthaptor indicated that this species very closely resembles G. salaris Malmberg, 1957. More detailed analysis revealed differences in the shape of the marginal hook sickle of these two species and thus Gyrodactylus teuchis Lautraite, Blanc, Thiery, Daniel & Vigneulle, 1999 was erected. Analysis of the ribosomal RNA gene or spacer sequences remains the most reliable method of identifying this species. This is believed to be the first record of a Gyrodactylus species identified first from molecular data and confirmed by morphological examination; previous molecular analyses had provided alternative methods for identifying species that had already been described using morphological characters.     

Gyrodactylus longidactylus n. sp., a monogenean from Pomatoschistus lozanoi (de Buen) (Gobiidae) from the North Sea

Gyrodactylus longidactylus n. sp., a gyrodactylid monogenean parasitising the gills of Pomatoschistus lozanoi (de Buen, 1923) in the North Sea is described. It is a species with rather small anchors which are only connected to a ventral bar which lacks a ventral bar membrane. The marginal hooks have long handles, which are always longer than the total length of the anchor. The pharynx has long pharyngeal processes. Measurements, drawings of the opisthaptoral hard parts, penis and pharynx, and SEM micrographs of this monogenean are presented. G. longidactylus n. sp. is the first monogenean species described from P. lozanoi.