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.     

Identification of a host-associated species complex using molecular and morphometric analyses,with the description of Gyrodactylus rugiensoides n. sp. (Gyrodactylidae,Monogenea)

Gyrodactylus rugiensis was originally described as a parasite occurring on the marine gobies Pomatoschistus minutus and Pomatoschistus microps. In our preliminary survey this species was also frequently found on Pomatoschistus pictus and Pomatoschistus lozanoi. Subsequent molecular analysis of the internal transcribed spacers rDNA region revealed that this parasite actually represents a complex of two apparently cryptic species, one restricted to P. microps and the other shared by P. minutus, P. lozanoi and P. pictus. Morphometric analyses were conducted on 17 features of the opisthaptoral hard parts of specimens collected from all four host species. Standard discriminant analysis showed a clear separation of both genotypes by significant differences in marginal hook and ventral bar features. Statistical classifiers (linear discriminant analysis and nearest neighbours) resulted in an estimated misclassification rate of 4.7 and 3.1%, respectively. Based on molecular, morphological and statistical analyses a new species, Gyrodactylus rugiensoides is described. This species seems to display a lower host-specificity than generally observed for Gyrodactylus species as it infects three sympatric host species.However, seasonal and host-dependent morphometric variation is shown for G. rugiensoides collected on P. pictus. Host-switching and gene flow might be important factors preventing speciation on closely related and sympatric host species. The presence of host associated species complexes in this Gyrodactylus-Pomatoschistus system is also confirmed by the presence of two host-dependent genotypes within G. micropsi found on P. minutus and P. lozanoi, and P. microps, respectively. By comparing host and parasite phylogeny, phylogenetic and ecological factors influencing host-specificity are discussed.     

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.     

New Macroderoides (Digenea: Macroderoididae) from Florida gar, with molecular phylogeny of the genus

Macroderoides minutus n. sp. is described based on specimens collected from the Florida gar (Lepisosteus platyrhincus) from Orange Lake, Alachua County, Florida. The new species is the ninth recognized species of Macroderoides and is morphologically closest to Macroderoides flavus and Macroderoides typicus. It differs from both species in having a smaller body, ventral sucker larger than the oral sucker, substantially lower body length to width ratio, fewer and larger eggs, and other characters. A comparison of sequences of nuclear rDNA (partial 18S, complete ITS region, and partial 28S) of the new species, with sequences of 5 other Macroderoides, convincingly supports the status of M. minutus as a new species. Molecular phylogenetic analysis, including all North American Macroderoides species except for Macroderoides parvus, has demonstrated the presence of 2 distinct clades, 1 including Macroderoides spinifer + (Macroderoides minutus n. sp. + Macroderoides texanus) and the other including Macroderoides trilobatus + (M. typicus + M. flavus). The tree topology suggests that the similarity in body proportions among M. minutus n. sp., M. typicus + M. flavus is not a result of close phylogenetic relationship. It has also revealed for the first time the phylogenetic affinities of M. flavus, which is the only North American member of Macroderoides that switched from holostean fishes to teleosts.     

Bravohollisia Bychowsky & Nagibina, 1970 and Caballeria Bychowsky & Nagibina, 1970 (Monogenea: Ancyrocephalidae) from Pomadasys hasta (Bloch) (Pomadasyidae), with the description of a new attachment mechanism

Nine species of monogeneans were collected from the fish Pomadasys hasta (Bloch): Bravohollisia magna Bychowsky & Nagibina, 1970; B. rosetta n. sp.; B. reticulata n. sp.; B. gussevi n. sp.; B. kritskyi n. sp.; Caballeria pedunculata Bychowsky & Nagibina, 1970; C. robusta Bychowsky & Nagibina, 1970; C. liewi n. sp.; and C. intermedius n. sp. The anchors of these species contain a canal beginning in the shaft and ending near the point; ducts of haptoral glands enter the canal in the shaft and are associated with net-like structures near the anchor tip. The net-like structures of Bravohollisia spp. are extensive, while those of Caballeria spp. are smaller and more compact. These nets probably assist the monogeneans in attaching to the gill filaments. Caballeria spp. possess four pairs of extensible haptoral digits. Species of Bravohollisia and Caballeria posses two seminal vesicles.     

Paleoclimatic history and vicariant speciation in the "sand goby" group (Gobiidae, Teleostei)

Vicariant and climatic cycling speciation hypotheses of the 'sand gobies' belonging to the genera Pomatoschistus, Gobiusculus, Knipowitschia, and Economidichthys are tested using molecular phylogenies constructed of nuclear DNA (ITS1 locus) and mitochondrial DNA (12S and 16S fragments). These gobies are among the most abundant in the Eastern Atlantic-Mediterranean region, and play an important role in the ecosystem. Considerable ITS1 length differences, primarily due to the presence of several tandem repeats, were found between species and even within individuals. Therefore, phylogenetic analyses focused on fragments of the 12S and 16S mtDNA region that have been sequenced for 16 goby taxa. The 'sand gobies' clustered as a monophyletic group as proposed on morphological grounds. However, G. flavescens, E. pygmaeus, and K. punctatissima clustered within the Pomatoschistus species, pointing to a paraphyletic origin of these genera. Furthermore, the genetic divergence between P. minutus from the Adriatic Sea versus the Atlantic-Mediterranean region was as high as the divergence within the P. minutus complex, suggesting that P. minutus from the Adriatic Sea should be considered as a distinct species. The "star" phylogeny might suggest that these gobies evolved in a very short time period, possibly linked to the drastic alterations in the Mediterranean Sea during and immediately after the Messinian salinity crisis at the end of the Miocene. The freshwater life-style appeared monophyletic; equating its origin with the salinity crisis resulted in a molecular clock estimate of 1.4% divergence per million years. The last common ancestor probably occupied sandy bottoms and a coastal niche while several species subsequently adapted to new habitats (pelagic, freshwater or stenohaline). The origin of the shallowest clades dated back to the glacial cycling during the Pleistocene epoch.     

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 five new Loma (Microsporidia) species in pacific fishes with redesignation of the type species Loma morhua Morrison & Sprague, 1981, based on morphological and molecular species-boundaries tests

Five new species of Loma were described from five Pacific fishes using light-microscopic and ultrastructural features along with phylogenetic analysis of the gene sequences of ribosomal RNA (rRNA) and elongation factor 1-alpha. Morphological data revealed both qualitative and quantitative differences in developmental stages and timing, vesicles, xenoma features, and spore sizes with statistical support that differentiated Loma pacificodae n. sp. in Pacific cod, Loma wallae n. sp. in walleye pollock, Loma kenti n. sp. in Pacific tomcod, Loma lingcodae n. sp. in lingcod, and Loma richardi n. sp. in sablefish from each other and other species in the genus. Phylogenetic analyses combined with monophyly tests supported species designations, but with low resolution in two cases perhaps due to rRNA paralogs or recent speciation. Loma branchialis in haddock was shown to be separate from Loma morhua in Atlantic cod, thereby making L. morhua, and not L. branchialis, the type species. A species from brook trout was shown to be a separate species from Loma salmonae, not a variant strain selected in the laboratory. By comparison with gadid host phylogeny, these Loma species appear to have coevolved with their hosts, first colonizing the Pacific basin about 12 million years ago.     

Three new species of Protogyrodactylus Johnston & Tiegs, 1922 (Monogenoidea: Dactylogyridae) from the gills of the longtail silverbiddy Gerres longirostris (Teleostei: Gerreidae) in the Red Sea

Twenty-one specimens of the longtailed silverbiddy Gerres longirostris (Gerreidae) were examined for dactylogyrid parasites from the Nabq Managed Resource Protected Area, Ras Mohammed National Park (Red Sea) near Sharm El-Sheikh, South Sinai, Egypt. The diagnosis of Protogyrodactylus Johnston & Tiegs, 1922 was amended, and three new species, P. federicae n. sp., P. zullinii n. sp. and P. alatus n. sp., were recovered and described; the prevalence of each species was 100%. P. federicae most closely resembled P. alienus Bychowsky & Nagibina, 1974, but differed from it by possessing two anteromedial projections on the ventral bar, a claw-like ventral anchor sclerite and spatulate dorsal bars. P. zullini was most similar to P. quadratus Johnston & Tiegs, 1922, from which it differed by having a distal hook on the superficial root of the dorsal anchor, an evenly curved ventral anchor shaft and point, and a flange on the bulbous base of the male copulatory organ. P. alatus was closest to P. youngi Bychowsky & Nagibina, 1974, from which it differed by having delicate anchors and two prominent anteromedial processes on the ventral bar.     

The parasite community infecting flounders, Platichthys flesus, in the tidal Thames.

The composition of the parasite fauna of the flounder, Platichthys flesus, retrieved from two locations in the tidal Thames is described in detail for the first time. The combined parasite species list of the flounders from Lots Road in the upper tideway and West Thurrock in the middle tideway consisted of one protozoan (Glugea stephani), one monogenean (Gyrodactylus sp.), four larval digeneans (Cryptocotyle concava, Timoniella imbutiforme, T. praeterita, and Labratrema minimus), five adult digeneans (Derogenes varicus, Lecithaster gibbosus, Podocotyle sp., Plagioporus varius, and Zoogonoides viviparus), one larval cestode (unidentified tetraphyllidean), one or possibly more larval nematodes (unidentified) plus five adult nematodes (Capillaria sp., Cucullanus heterochrous, C. minutus, Contracaecum sp. and Goezia sp.), two acanthocephalans (Pomphorhynchus laevis and Acanthocephalus anguillae), three copepods (Lepeophtheirus pectoralis, Acanthochondria sp. and Lernaeocera branchialis), and one mollusc (unidentified glochidia). The overall parasite community of flounders from Lots Road and West Thurrock were compared in terms of species richness and diversity. The parasite community in flounders from the former location in the upper tideway was found to be less diverse than that of its counterpart at West Thurrock in the middle estuary. The component community of Lots Road flounders was dominated by the acanthocephalan Pomphorhynchus laevis.     

A new goby, Glossogobius minutus (Teleostei: Gobiidae) from the south-west coast of India

A new species of gobiid fish Glossogobius minutus collected from the south-west coast of India is described. The morphological, morphometric and meristic characters were studied with special emphasis on osteology. The various characters are compared with those of G. giuris (Harn.) and G. biocellatus (Val.), the only other species of the genus reported from this region. Glossogobius minutus is a benthic omnivore and shows distinct sexual dimorphism.     

Taxonomy and host-specificity of Gyrodactylus aideni n. sp. and G. pleuronecti (Monogenea: Gyrodactylidae) from Pseudopleuronectes americanus (Walbaum) in Passamaquoddy Bay, New Brunswick, Canada

Wild and cultured winter flounder Pseudopleuronectes americanus (Walbaum) from Passamaquoddy Bay were surveyed for species of Gyrodactylus Nordmann, 1832. Two species were found: G. pleuronecti Cone, 1981 and G. aideni n. sp, both members of Malmberg’s ‘groenlandicus group’. Although the hard parts in the haptor are very similar in the two species, hamuli of G. aideni are consistently shorter than those of G. pleuronecti. The two species differed by 35 base pairs in the ITS 1, 5.8 and ITS 2 region. A BLAST search identified a variety of species of Gyrodactylus from marine fishes in the Atlantic Ocean as closest matches, indicating the ‘groenlandicus group’ is part of a major marine lineage within Gyrodactylus (sensu lato) that has successfully radiated among coastal percid, pleuronectid, cottid and anarhichadid fishes. Exposure experiments suggested that winter flounder is the primary host of both species of parasites and that three other pleuronectid species in the bay may potentially serve only as occasional transport hosts.     

Description of two new species of Glypthelmins Stafford, 1905 (Digenea: Macroderoididae) in Rana spp. from Mexico, based on morphology and mtDNA and rDNA sequences

Glypthelmins Stafford, 1905 includes 29 putative species commonly found in the intestine and liver of anurans from all over the world but mainly in the Americas. Partial sequences of the cytochrome c oxidase subunit 1 ( cox 1), ribosomal internal transcribed spacer region 2 (ITS2) and the large subunit 28S rDNA gene were obtained and analysed using pairwise distance matrices and parsimony methods in order to characterise the interrelationships between 14 isolates of four nominal species of Glypthelmins recognised on morphological grounds. The highest intra-specific sequence divergence occurred in the cox 1 (18.53%) sequence, followed by that of the ITS2 (5.44%) and 28S (4.63%). Genetic variability was detected between the three isolates originally identified as G. facioi Brenes et al., 1959 from two localities in Mexico and one locality in Costa Rica. Sequence divergence exhibited among these isolates ranged from 10.70 to 11.22%, from 0.48 to 0.97% and from 1.33 to 1.88% for cox 1, ITS2 and 28S, respectively. Phylogenetic analysis combining all three data-sets generated a single most parsimonious tree. The three isolates of G. facioi form a clade, with an isolate collected from frogs in Veracruz State as the sister group to an isolate from Tabasco State + G. facioi from Costa Rica. The information derived from pairwise distance of independent data-sets plus the phylogenetic information indicate that each of the two isolates from Mexico, identified a priori as G. facioi, represent separate species. A re-examination of specimens was carried out and a re-evaluation made of the morphological characters to find reliable differences that had been overlooked. As a consequence, G. brownorumae n. sp. from Tabasco and G. tuxtlasensis n. sp. from Veracruz are described based on molecular and morphological differences.     

Molecular faunistics of accidental infections of <Emphasis Type="Italic">Gyrodactylus</Emphasis> Nordmann, 1832 (Monogenea) parasitic on salmon <Emphasis Type="Italic">Salmo salar</Emphasis> L. and brown trout <Emphasis Type="Italic">Salmo trutta</Emphasis> L. in NW Russia

Salmon Salmo salar L. and brown trout S. trutta L. juveniles were examined for the presence of accidental monogenean ectoparasitic species of Gyrodactylus Nordmann, 1832 in the Baltic and White Sea basins of Russian Karelia in order to estimate the frequency of host-switching attempts on an ecological timescale. To collect phylogeographical information and for exact species identification, the parasites were characterised by nuclear internal transcribed spacer sequences of rDNA (ITS) and, for some species, also by their mitochondrial DNA (CO1 gene) sequences. Four accidental Gyrodactylus species were observed on salmon and brown trout. A few specimens of G. aphyae Malmberg, 1957, the normal host of which is the Eurasian minnow Phoxinus phoxinus (L.), were observed on lake salmon from the Rivers Kurzhma (Lake Kuito, White Sea basin) and Vidlitsa (Lake Ladoga, Baltic basin). G. lucii Kulakovskaya, 1952, a parasite of the northern pike Esox lucius L., was observed on salmon in the Kurzhma. In the River Vidlitsa, two specimens of G. papernai Ergens & Bychowsky, 1967, normally on stone loach Barbatula barbatula (L.), were found on salmon. On anadromous White Sea salmon in the River Pulonga in Chupa Bay, a few salmon parr carried small colonies of G. arcuatus Bychowsky, 1933, which were shown to have originated from the local three-spined stickleback Gasterosteus aculeatus L. consumed as prey. No specimens of Gyrodactylus salaris Malmberg, 1957 were observed, although the Pulonga is the nearest salmon spawning river to the River Keret', which is heavily infected with introduced G. salaris. In the River Satulinoja, Lake Ladoga, three specimens of G. lotae Gusev, 1953, from burbot Lota lota (L.), were collected from a single brown trout S. trutta. All nonspecific gyrodactylid infections on salmonids were judged to be temporary, because only a few specimens were observed on each of the small number of infected fishes. The prevalence of endemic G. salaris was also low, only 1% (Nfish = 296) in Lake Onega and 0.7% (Nfish = 255) in Lake Ladoga, while brown trout specific Gyrodactylus species were not observed on any of the 429 trout examined from the Ladoga basin. The host-specific and unspecific burden of Gyrodactylus spp. on these 'glacial relict' populations of salmon and brown trout was very low, suggesting a generalised resistance against the co-evolved freshwater parasite community, or some kind of 'vaccination' effect. These hypotheses deserve further testing.     

Biochemical polymorphisms and phenetic relationships in rodents of the genusCtenomys from Southern Brazil

Protein variation at 20 loci was analyzed by starch gel electrophoresis in population samples of Ctenomys flamarioni, C. torquatus, C. sp., and C. minutus collected in 25 localities of Southern Brazil. Results show that these four species exhibit higher levels of genetic variability (He = 0.11-0.17) than those reported for most other fossorial rodents. Estimates of similarity coefficients indicate that C. minutus and C. sp. (S = 0.91) are the closest species, while C. flamarioni (S = 0.77) is the most distant from the others. The data presented here support the hypothesis of a relatively recent disjunction of C. minutus from C. sp. Within-species similarity indices are of the same order of magnitude as those found between species. The relatively high levels of heterozygosity observed are not in accordance with the view that subterranean taxa should be less variable than aboveground species due to the uniformity of their environment.     

Gyrodactylus longipes n. sp. (Monogenea: Gyrodactylidae) from farmed gilthead seabream (Sparus aurata L.) from the Mediterranean

Gyrodactylus longipes n. sp. (Monogenea, Gyrodactylidae) is described from the gills of farmed juvenile gilthead seabream (Sparus aurata L.) from two sites located in Italy and Bosnia-Herzegovina and represents the second species of Gyrodactylus to be described from S. aurata. Gyrodactylus orecchiae Paladini, Cable, Fioravanti, Faria, Di Cave et Shinn, 2009 was the first gyrodactylid to be described from S. aurata, from populations cultured in Albania and Croatia. In the current study, G. longipes was found in a mixed infection with G. orecchiae on fish maintained in Latina Province, Italy, thus extending the reported distribution of the latter throughout the Mediterranean. The morphology of the opisthaptoral hard parts of G. longipes is compared to those of G. orecchiae, using light and scanning electron microscopy. Gyrodactylus longipes is characterised by having larger, elongated ventral bar processes and long, triangular-shaped toe region to their marginal hook sickles which, by comparison, are rhomboid in G. orecchiae. The marginal hook sickles of G. longipes are almost double the size of G. orecchiae which allows for their rapid discrimination from each other in mixed infections. A comparison of the DNA sequence of the ribosomal internal transcribed spacer 1 and 2 regions (ITS1 and ITS2) of G. longipes with the corresponding sequence from G. orecchiae and with those available in GenBank, supports the separate species status of G. longipes. Part of this study necessitated an overview of the existing Gyrodactylus fauna from Italy and Bosnia-Herzegovina; a summary from each country is provided here to assist future investigations.     

Phylogenetic analysis of a free-living strain of Symbiodinium isolated from Jiaozhou Bay, P.R. China

This paper reported the isolation and the phylogenetic analysis of a free-living Gymnodinium-like dinoflagellates (G. sp. (15)) from the sea water collected at Jiaozhou bay, Qingdao, Shandong Province, P.R. China. Phylogenetic reconstruction analysis with Neighbor-Joining (NJ) method using sequences of variable regions (V1+V2+V3) of the small subunit (SSU) rDNA indicated that G. sp. (15) was a Symbiodinium sp., which was closely related to a symbiont of anemones, S. californium, and the free-living strain, Gymnodinium varians. These three strains formed a new clade (which had been designated as clade F) with 100% bootstrap support. Sequence comparison showed that sequences of the internal transcribed spacer (ITS) of these three strains were highly homologous, suggesting that they might belong to one species.