Dracunculoid nematodes (Spirurida: Dracunculoidea) of fishes from the Volga River delta

Faunistic and some morphological data, as well as nomenclature notes on dracunculoid nematodes parasitising fishes in the Volga River delta, are presented. The author replaced a preoccupied generic name Molnaria Moravec, 1968 (Skrjabillanidae) by the new name Kalmanmolnaria nom. nov. The validity of Philometroides lusii (Vismanis, 1962) comb. nov. as a senior objective synonym of Philometroides lusiana (Vismanis, 1967) Ivaschkin et al., 1971 is restored.     

Morphology of Philometroides barbi (Nematoda: Philometridae), a rare tissue parasite of the Mediterranean barbel Barbus meridionalis (Osteichthyes)

A female specimen of the nematode Philometroides barbi Moravec, Simková, Hanzelová, Spakulová et Caki?, 2005, a little-known histozoic parasite of the Mediterranean Barbus meridionalis, was recorded from the fin of its fish host in Bulgaria. Scanning electron microscopical examination, used for the first time in this species, made it possible to determine the character of cephalic papillae (14 papillae arranged in 2 circles) and amphids and confirmed the presence of 4 large caudal lobes. These features distinctly distinguish P. barbi from Philometroides cyprini and other congeners parasitizing European fishes.     

Three new species of philometrids (Nematoda: Philometridae) from centrarchid fishes in the USA

The following 3 new species of the Philometridae (Nematoda: Dracunculoidea) are described from freshwater centrarchid fishes (Centrarchidae: Perciformes) from eastern North America on the basis of museum and newly collected specimens: Philometra orbitalensis n. sp. and Philometroides aphanonaris n. sp. from the oculo-orbits and subcutaneous tissues of the head, respectively, of the largemouth bass Micropterus salmoides, and Philometroides wellborni n. sp. from the oculo-orbits of the bluegill Lepomis macrochirus (type host) and the redbreast sunfish Lepomis auritus. Whereas P. wellborni is described from both males and gravid females, the males of P. aphanonaris and P. orbitalensis remain unknown. The type locality of all 3 species is the West Point Reservoir, Alabama-Georgia; P. aphanonaris and P. wellborni have also been recorded from the Santee River in South Carolina. In contrast to other Philometra spp. parasitizing North American freshwater fishes, the gravid females of P. orbitalensis are characterized by large cephalic papillae of the external circle, yellowish body color, location (oculo-orbits) in the host, and by the host type (Centrarchidae). Philometroides aphanonaris and P. wellborni differ from North American congeners from freshwater fishes in the absence of esophageal teeth or a different embossment of the caudal end in gravid females; P. wellborni differs from P. aphanonaris in the absence of cuticular bosses from the caudal end of gravid females and in some other features (extent of embossment, body color, location, and host type).     

New observations on philometrid nematodes (Philometridae) in marine fishes from the Northern Gulf of Mexico and the Indian River Lagoon of Florida (Usa), with first description of the male of Caranginema americanum

The following 3 species of Philometra Costa, 1845 (Nematoda: Philometridae) were recorded from marine fishes off Florida: Caranginema americanum Moravec, Montoya-Mendoza and Salgado-Maldonado, 2008 from the subcutaneous tissue of the crevalle jack Caranx hippos (Linnaeus) (Carangidae); Philometra charlestonensis Moravec, de Buron, Baker and González-Solís, 2008 from the gonads (ovaries) of the scamp Mycteroperca phenax Jordan and Swain (Serranidae); and Philometra sp. (only subgravid females) from the gonads (ovaries) of the Atlantic needlefish Strongylura marina (Walbaum) (Belonidae). The male of C. americanum , the type species of Caranginema Moravec, Montoya-Mendoza, and Salgado-Maldonado, 2008 , is described for the first time. Its general morphology is similar to that of males of Philometra and Philometroides species. The males of C. americanum are mainly characterized by an elongate body, 3.13-3.28 mm long, a markedly elongate esophagus, and spicules and a gubernaculum 69-75 μm and 48-51 μm long, respectively. The present findings of C. americanum and P. charlestonensis represent new geographical records. The gonad-infecting Philometra sp. from S. marina probably belongs to an undescribed species.     

Observations on five species of philometrid nematodes from marine fishes in Japan

Five species of philometrid nematodes (subfamily Philometrinae) are redescribed on the basis of newly collected materials from marine fishes from Japan. The species Philometra sciaenae Yamaguti, 1941, considered by Rasheed (1963) a synonym of P. lateolabracis (Yamaguti, 1935), is re-established as a valid species and the male of this nematode, collected from the ovary of the type-host, Argyrosomus argentatus, is described for the first time; the latter is characterised mainly by the length of equally long spicules (0.108–0.126 mm) and the length ratio of the gubernaculum and spicules (1:2.25). Other species redescribed on females include Philometra inimici Yamaguti, 1941 and P. sebastisci Yamaguti, 1941 from the ovary of their type-hosts, Inimicus japonicus and Sebastiscus marmoratus, respectively, and also Philometroides seriolae (Ishii, 1931) from the musculature of Seriola quinqueradiata (type-host), and Clavinema mariae (Layman, 1930) from the subcutaneous tissue of Pleuronectes schrenki and Acanthogobius flavimanus (a new host record). For the first time, cephalic ends of some of these species were studied under the scanning electron microscope (SEM) and the character of their cephalic papillae was described. The importance of a knowledge of male morphology for the taxonomy of philometrids is stressed.     

Evolutionary relationships of Spirurina (Nematoda: Chromadorea: Rhabditida) with special emphasis on dracunculoid nematodes inferred from SSU rRNA gene sequences

The analysis of 26 new small subunit rRNA sequences obtained from helminths that primarily parasitize fishes sampled from five continents provided well-supported trees, allowing us to study the phylogenetic relationships among spirurid nematodes. The analyses have shown that Dracunculoidea is a paraphyletic taxon and Anguillicolidae and Gnathostomatidae constitute the basal branch of the suborder Spirurina. The genera Philometra and Philometroides appear to be paraphyletic, while on the higher taxonomic level, good correlation between the morphology-based system and molecular data was observed. Neither co-evolution of the studied helminths with their hosts, nor phylogeographic pattern, are apparent in our dataset.     

中国9种嗜子宫线虫系统发育的初步研究

为了探讨鱼类寄生嗜子宫线虫的系统发育关系,测定了8种嗜子宫线虫的ITS rDNA(核糖体转录内间隔区核 糖核酸)序列和9种嗜子宫线虫的18S rDNA(小亚基核糖体核糖核酸)部分序列,并构建了18S rDNA序列的系统发 育树。在比较和分析ITS rDNA和18S rDNA两种分子标记对嗜子宫科线虫系统发育适用性的基础上,分析了嗜子 宫线虫的系统发育关系。结果表明:中国嗜子宫线虫是单系起源;黄颡鱼似嗜子宫线虫、赣州似嗜子宫线虫和棍头 嗜子宫线虫亲缘关系非常接近,可能是较晚形成的种;似嗜子宫线虫属可能应该被细分为更多的属。       

Neophilometroides n. gen. (Nematoda: Philometridae) for Philometroides caudatus Moravec,Scholz and Vivas-Rodríguez, 1995, with erection of Neophilometroidinae n. subfam

A detailed examination (including scanning electron microscopy) of newly collected nematode specimens (subgravid females) referable to Philometroides caudatus Moravec, Scholz and Vivas-Rodríguez, 1995, from the swimbladder of the pimelodid catfish, Rhamdia guatemalensis, in the Papaloapan River in Tlacotalpan, State of Veracruz, Mexico, as well as the paratype specimens of this species, revealed the presence of a small buccal capsule armed with peribuccal teeth in the female. Because of this unique morphological feature within the Philometridae, and considering other taxonomically important characters such as the structure of the male tail and the shape of the anterior end of the esophagus in the gravid female, Neophilometroidinae n. subfam, and Neophilometroides n. gen. are proposed to accommodate this species. Philometra paraguayensis Petter, 1995, is transferred to Neophilometroides as N. paraguayensis (Petter, 1995) n. comb. Newly collected specimens are briefly described and illustrated.     

A hyperosmotic stress-induced mRNA of carp cell encodes Na(+)- and Cl(-)-dependent high affinity taurine transporter

A cDNA clone encoding a Na(+)- and Cl(-)-dependent high affinity taurine transporter was isolated from a common carp cell line, Epithelioma papulosum cyprini (EPC), as a hyperosmotic stress-inducible gene by RNA arbitrarily primed PCR. The clone contained a 2.5-kb cDNA fragment including an open reading frame of 1878 bp encoding a protein of 625 amino acids. The deduced amino acid sequence of carp taurine transporter shows 78-80% identity to those of cloned mammalian taurine transporters. The functional characteristics of the cloned transporter were analyzed by expression in COS-7 cells. Transfection with the cDNA induced Na(+)- and Cl(-)-dependent taurine transport activity with an apparent K(m) of 56 microM. The Na(+)/Cl(-)hepatopancreas. Taurine transporter mRNA level increased up to 7.5-fold on raising the ambient osmolality from 300 to 450 mosmol/kgH(2)O. These data suggest the significant role of taurine as an osmolyte in carp cells.     

Philometrids of the southern flounder Paralichthys lethostigma: a multidimensional approach to determine their diversity

Two species of philometrid nematode, Philometra overstreeti and Philometroides paralichthydis, infect the southern flounder, Paralichthys lethostigma. Individuals of P. overstreeti are located between the teeth and inside the bony part of the branchial arches of the fish. Individuals of P. paralichthydis are associated with the bones of the buccal cavity and among muscles that control the dorsal and anal fins. Sequencing of part of the cytochrome oxidase I gene revealed 4 distinct genetic clades, each corresponding exactly to the 4 respective locations of the parasites in the host, suggesting the need for taxonomic revision. We hypothesized that each clade represented a separate species and, because the worms are morphologically indistinguishable, compared population level parameters of the clades comprising each currently recognized species. For each currently recognized species, the presence of worms from 1 clade was negatively correlated with the presence of worms from the other. Results also indicated significant differences between the clades in prevalences relative to both biotic and abiotic factors. Results clearly indicated major differences in the ecology of the philometrids constituting each clade. Taken as a whole, molecular and ecological data support the contention that the 4 genetic clades are likely 4 distinct species.     

A permanent fish cell line (EPC) for genotoxicity testing of marine sediments with the comet assay.

Genotoxicity and cytotoxicity were evaluated in an in vitro system with a permanent cell line Epithelioma papulosum cyprini (EPC) derived from a skin tumour of carp (Cyprinus carpio L.). EPC cells were exposed to different concentrations of organic sediment extracts from the North Sea for 24h. After incubation the cells were analysed for viability and DNA strand breaks with the comet assay or single cell gel electrophoresis (SCGE). The results confirm the sensitivity of this assay. Out of 10 marine sediment samples from the North Sea, 9 showed a dose-dependent genotoxic effect. The EC(50) of sediment extracts ranged from 7 to 307 mg sediment dry weight/ml assay volume. Hepatic microsomal enzymes from dab (Limanda limanda L.) was proposed for enzymatic activation of benzo[a]pyrene (BAP) or sediment extracts, respectively. The suitability of this in vitro test system for assessing genotoxic and cytotoxic effects of marine sediment extracts on EPC cells could be demonstrated.     

丹江口水库鲤肠道寄生蠕虫群落结构与季节动态

2004年2月到2005年11月在丹江口水库库区206尾鲤(Cyprinus carpio)肠道中检获蠕虫11种,其中复殖吸虫3种,线虫5种,棘头虫2种,绦虫1种。总体感染率为45.63%,平均感染丰度为4.23±12.65,平均感染强度为9.29±17.48,其中饭岛盾腹吸虫(Aspidogaster ijimai)的总感染率(25.24%)和平均感染丰度(1.76±6.46)最大,瓣睾鲫吸虫(Carassatrema lamellorchis)的感染强度(25.00±46.68)最大。除部分平均感染丰度较低的线虫如鲤带巾线虫(Cucullanus cyprini)外,其他蠕虫的分布类型均为聚集分布,蠕虫群落多样性指数为4.63,均匀度指数为0.60,对群落多样性的季节动态分析表明,各季节群落多样性和均匀度波动较大,并无明显变化规律。每尾鲤感染蠕虫种数多在1—4种之间,所有感染的11种蠕虫中优势种为饭岛盾腹吸虫;次优势种为日本侧殖吸虫(Asymphylodora japonica)、中华许氏绦虫(Khawia sinensis)、瓣睾棘吸虫和鲤长棘吻虫(Rhadinarhynchus cyprini);非优势种为对盲囊线虫(Contracaecum sp.)、鲤带巾线虫、鲤杆咽吸虫(Rhabdochona cyprini)、黄颡刺盖线虫(Spinitectus gigi)、毛细线虫(Capillaria sp.)和木村小棘吻虫(Micracanthorhynchina motomurai)。在种间协调关系方面,鲤杆咽线虫和瓣睾鲫吸虫、鲤长棘吻虫和饭岛盾腹吸虫、对盲囊线虫和木村小棘吻虫、鲤长棘吻虫和木村小棘吻虫之间分别存在显著正关联。对优势种和次优势种蠕虫中种群的季节动态分析表明,鲤寄生蠕虫各组分的感染率和平均感染丰度存在显著的季节差异,在秋、冬季节的感染水平普遍比较高,而到春夏则急剧下降,但中华许氏绦虫无显著季节变化。     

Seasonal development and pathological changes associated with the parasitic nematode Philometroides sanguineus in wild crucian carp Carassius carassius (L.) in England

Pathological changes associated with the parasitic nematode Philometroides sanguineus (Rudolphi, 1819) are described for the first time from observations of infections in wild crucian carp Carassius carassius (L.) in England. The damage caused by P. sanguineus was influenced strongly by host size, parasite development and the seasonal migrations of female nematodes within host tissues. Male and unfertilized female nematodes located on the swim-bladder wall caused only minor, localized changes. In contrast, the migration of gravid female nematodes into the fins during autumn provoked an acute inflammatory response comprising neutrophils, eosinophils and lymphocytes. This was accompanied by fin distortion, swelling of the dorsal and caudal tissues, degenerative changes and localized oedema. The encapsulation of female nematodes in connective tissue throughout winter limited additional tissue damage. The emergence of gravid nematodes from the fins in spring to facilitate larval dispersal caused necrosis, hyperplasia and loss of skin integrity. This activity was again accompanied by acute inflammatory reactions. Pathological changes were more severe in crucian carp measuring less than 60 mm in length, with no signs of debilitation in larger fish. These observations suggest that any impact of P. sanguineus is strictly seasonal and may be limited to fry. Lesions caused by this parasite, only recently recorded in Britain, may represent an additional pressure upon wild crucian carp populations already threatened by hybridization, competition and habitat loss.     

Description of an as yet unclassified DNA virus from diseased Cyprinus carpio species

Numerous deaths of koi and common carp (Cyprinus carpio) were observed on many farms throughout Israel, resulting in severe financial losses. The lethal viral disease observed is highly contagious and extremely virulent, but morbidity and mortality are restricted to koi and common carp populations. Diseased fish exhibit fatigue and gasping movements in shallow water. Infected fish had interstitial nephritis and gill necrosis as well as petechial hemorrhages in the liver and other symptoms that were not consistent with viral disease, suggesting a secondary infection. Here we report the isolation of carp nephritis and gill necrosis virus (CNGV), which is the etiologic agent of this disease. The virus propagates and induces severe cytopathic effects by 5 days postinfection in fresh koi or carp fin cell cultures (KFC and CFC, respectively), but not in epithelioma papillosum cyprini cells. The virus harvested from KFC cultures induced the same clinical signs, with a mortality of 75 to 95%, upon inoculation into naive koi and common carp. Using PCR, we provide final proof that the isolated virus is indeed the etiologic agent of food and ornamental carp mortalities in fish husbandry. Electron microscopy revealed viral cores with icosahedral morphology of 100 to 110 nm that resembled herpesviruses. Electron micrographs of purified pelleted CNGV sections, together with viral sensitivities to ether and Triton X-100, suggested that it is an enveloped virus. However, the genome of the isolated virus is a double-stranded DNA (dsDNA) molecule of 270 to 290 kbp, which is larger than known herpesviruses. The viral DNA seems highly divergent and bears only small fragments (16 to 45 bp) that are similar to the genomes of several DNA viruses. Nevertheless, amino acid sequences encoded by CNGV DNA fragments bear similarities primarily to members of the Poxviridae and Herpesviridae and to other large dsDNA viruses. We suggest, therefore, that the etiologic agent of this disease may represent an as yet unclassified virus species that is endemic in C. carpio (carp).     

A permanent fish cell line (EPC) for genotoxicity testing of marine sediments with the comet assay

Genotoxicity and cytotoxicity were evaluated in an in vitro system with a permanent cell line Epithelioma papulosum cyprini (EPC) derived from a skin tumour of carp (Cyprinus carpio L.). EPC cells were exposed to different concentrations of organic sediment extracts from the North Sea for 24 h. After incubation the cells were analysed for viability and DNA strand breaks with the comet assay or single cell gel electrophoresis (SCGE). The results confirm the sensitivity of this assay. Out of 10 marine sediment samples from the North Sea, 9 showed a dose-dependent genotoxic effect. The EC₅₀ of sediment extracts ranged from 7 to 307 mg sediment dry weight/ml assay volume. Hepatic microsomal enzymes from dab (Limanda limanda L.) was proposed for enzymatic activation of benzo[a]pyrene (BAP) or sediment extracts, respectively. The suitability of this in vitro test system for assessing genotoxic and cytotoxic effects of marine sediment extracts on EPC cells could be demonstrated.     

Isolation of a rhabdovirus during outbreaks of disease in cyprinid fish species at fishery sites in England

A virus was isolated during disease outbreaks in bream Abramis brama, tench Tinca tinca, roach Rutilis rutilis and crucian carp Carassius carassius populations at 6 fishery sites in England in 1999. Mortalities at the sites were primarily among recently introduced fish and the predominant fish species affected was bream. The bream stocked at 5 of the 6 English fishery sites were found to have originated from the River Bann, Northern Ireland. Most fish presented few consistent external signs of disease but some exhibited clinical signs similar to those of spring viraemia of carp (SVC), with extensive skin haemorrhages, ulceration on the flanks and internal signs including ascites and petechial haemorrhages. The most prominent histopathological changes were hepatocellular necrosis, interstitial nephritis and splenitis. The virus induced a cytopathic effect in tissue cultures (Epithelioma papulosum cyprini [EPC] cells) at 20 degrees C and produced moderate signals in an enzyme immunoassay (EIA) for the detection of SVC virus. The virus showed a close serological relationship to pike fry rhabdovirus in both EIA and serum neutralisation assays and to a rhabdovirus isolated during a disease outbreak in a bream population in the River Bann in 1998. A high degree of sequence similarity (> or = 99.5% nucleotide identity) was observed between the English isolates and those from the River Bann. Experimental infection of juvenile bream, tench and carp with EPC cell-grown rhabdovirus by bath and intraperitoneal injection resulted in a 40% mortality of bream in the injection group only. The virus was re-isolated from pooled kidney, liver and spleen tissue samples from moribund bream. The field observations together with the experimental results indicate that this rhabdovirus is of low virulence but may have the potential to cause significant mortality in fishes under stress.     

Skrjabillanus cyprini n. sp. (Nematoda: Dracunculoidea) from the scales of common carp Cyprinus carpio (Pisces) from Hungary

A new species of the histozoic nematode, Skrjabillanus cyprini n. sp., is described from the subsquamal part of the scales of common carp Cyprinus carpio L. from Lake Balaton, Hungary. It differs from the other three congeneric species mainly in possessing an extremely small, oval-shaped buccal capsule with a large tooth at the bottom, in the shape of the cephalic end (somewhat bulbously inflated, truncated anteriorly), the extent of the oesophageal glands and body size (body length of male and female 1,800–3,000 µm and 3,200–4,100 µm, respectively). A key to the identification of Skrjabillanus spp. is provided.