Molecular Identification of Anisakis Larvae Extracted by Gastrointestinal Endoscopy from Health Check-up Patients in Korea.

Anisakiasis is a zoonotic disease induced by anisakid nematodes, and endoscopic inspection is used for a diagnosis or remedy for it. Anisakis simplex, Anisakis physeteris, and Pseudoterranova decipiens had been reported to be the major species causing human infections, particularly, in Japan. However, in Korea, recent studies strongly suggested that Anisakis pegreffii is the major species of human infections. To support this suggestion, we collected anisakid larvae (n=20) from 20 human patients who were undergone gastrointestinal endoscopy at a health check-up center in Korea, and molecular identification was performed on the larvae using PCR-RFLP analysis and gene sequencing of rDNA ITS regions and mtDNA cox2. In addition, anisakid larvae (n=53) collected from the sea eel (Astroconger myriaster) were also examined for comparison with those extracted from humans. The results showed that all human samples (100%) were identified as A. pegreffii, whereas 90.7% of the samples from the sea eel were A. pegreffii with the remaining 9.3% being Hysterothylacium aduncum. Our study confirmed that A. pegreffii is the predominant species causing human anisakiasis in Korea, and this seems to be due to the predominance of this larval type in the fish (sea eels) popularly consumed by the Korean people. The possibility of human infection with H. aduncum in Korea is also suggested.

Studies of Myxidium giardi Cépède, 1906 infections in Icelandic eels identifies a genetically diverse clade of myxosporeans that represents the Paramyxidium n. g. (Myxosporea: Myxidiidae).

BACKGROUND: The myxosporean Myxidium giardi Cépède, 1906 was described infecting the kidney of the European eel, Anguilla anguilla (L.), having spindle-shaped myxospores and terminal sub-spherical polar capsules. Since then, numerous anguillid eels globally have been documented to have similar Myxidium infections. Many of these have been identified using the morphological features of myxospores or by the location of infection in the host, and some have been subsequently synonymised with M. giardi. Therefore, it is not clear whether M. giardi is a widely distributed parasite, infecting numerous species of eels, in multiple organs, or whether some infections represent other, morphologically similar but different species of myxosporeans. The aim of the present study was to assess the status of M. giardi infections in Icelandic eels, and related fish hosts in Malaysia and to use spore morphology and molecular techniques to evaluate the diversity of myxosporeans present. RESULTS: The morphologies of the myxospores from Icelandic eels were very similar but the overall dimensions were significantly different from the various tissue locations. Myxospores from the kidney of the Malaysian tarpon, Megalops cyprinoides (Broussonet), were noticeably smaller. However, the SSU rDNA sequences from the different tissues locations in eels, were all very distinct, with percentage similarities ranging from 92.93% to as low as 89.8%, with the sequence from Malaysia being even more dissimilar. Molecular phylogenies consistently placed these sequences together in a clade that we refer to as the Paramyxidium clade that is strongly associated with the Myxidium clade (sensu stricto). We erect the genus Paramyxidium n. g. (Myxidiidae) to accommodate these histozoic taxa, and transfer Myxidium giardi as Paramyxidium giardi Cépède, 1906 n. comb. as the type-species. CONCLUSIONS: There is not a single species of Myxidium (M. giardi) causing systemic infections in eels in Iceland. There are three species, confirmed with a robust phylogeny, one of which represents Paramyxidium giardi n. comb. Additional species probably exist that infect different tissues in the eel and the site of infection in the host fish is an important diagnostic feature for this group (Paramyxidium n. g. clade). Myxospore morphology is generally conserved in the Paramyxidium clade, although actual spore dimensions can vary between some species. Paramyxidium spp. are currently only known to infect fishes from the Elopomorpha.

Three new species of Cucullanus (Nematoda: Cucullanidae) from marine fishes off New Caledonia, with a key to species of Cucullanus from Anguilliformes.

Based on light and scanning electron microscopical studies of nematode specimens from the digestive tract of some rarely collected anguilliform and perciform fishes off New Caledonia, three new species of Cucullanus Müller, 1777 (Cucullanidae) are described: C. austropacificus n. sp. from the longfin African conger Conger cinereus (Congridae), C. gymnothoracis n. sp. from the lipspot moray Gymnothorax chilospilus (Muraenidae), and C. incognitus n. sp. from the seabream Dentex fourmanoiri (Sparidae). Cucullanus austropacificus n. sp. is characterized by the presence of cervical alae, ventral sucker, alate spicules 1.30-1.65 mm long, conspicuous outgrowths of the anterior and posterior cloacal lips and by elongate-oval eggs measuring 89-108 × 48-57 µm; C. gymnothoracis n. sp. is similar to the foregoing species, but differs from it in the absence of cervical alae and the posterior cloacal outgrowth, in the shape and size of the anterior cloacal outgrowth and somewhat shorter spicules 1.12 mm long; C. incognitus n. sp. (based on female morphology) differs from other congeneric species parasitic in the Sparidae mainly in possessing cervical alae, the postequatorial vulva, phasmids situated at the mid-length of the tail and in the size of the eggs (75-84 × 45-66 µm). A key to species of Cucullanus parasitizing anguilliform fishes is provided.

TITLE: Trois nouvelles espèces de Cucullanus (Nematoda: Cucullanidae) de poissons marins au large de la Nouvelle-Calédonie, avec une clé des espèces de Cucullanus d'Anguilliformes. ABSTRACT: Sur la base d'une étude en microscopie photonique et électronique à balayage de spécimens de nématodes provenant du tube digestif de quelques poissons anguilliformes et perciformes rarement pêchés en Nouvelle-Calédonie, trois nouvelles espèces de Cucullanus Müller, 1777 (Cucullanidae) sont décrites : C. austropacificus n. sp. de Conger cinereus (Congridae), C. gymnothoracis n. sp. de Gymnothorax chilospilus (Muraenidae) et C. incognitus n. sp. de Dentex fourmanoiri (Sparidae). Cucullanus austropacificus n. sp. est caractérisé par la présence d'ailes cervicales, une ventouse ventrale, des spicules ailés de 1,30-1,65 mm de long, des excroissances remarquables des lèvres cloacales antérieure et postérieure et d'oeufs ovales et allongés mesurant 89-108 × 48-57 µm. C. gymnothoracis n. sp. est semblable à l'espèce précédente, mais en diffère en l'absence d'ailes cervicales et d'excroissance cloacale postérieure, par la forme et la taille de l'excroissance cloacale antérieure et par des spicules un peu plus courts de 1,12 mm. C. incognitus n. sp. (basé sur la morphologie de la femelle) diffère des autres espèces congénères parasites chez les Sparidae principalement par la possession d'ailes cervicales, une vulve postéquatoriale, des phasmides situés à mi-longueur de la queue et par la taille des oeufs (75­84 × 45­66 µm). Une clé des espèces de Cucullanus parasitant les poissons anguilliformes est fournie.

The Influence of Sex, Parasitism, and Ontogeny on the Physiological Response of European Eels (Anguilla anguilla) to an Abiotic Stressor.

Migration of adult European eels (Anguilla anguilla) from freshwater feeding grounds to oceanic spawning grounds is an energetically demanding process and is accompanied by dramatic physiological and behavioral changes. Humans have altered the aquatic environment (e.g., dams) and made an inherently challenging migration even more difficult; human activity is regarded as the primary driver of the collapse in eel populations. The neuroendocrine stress response is central in coping with these challenging conditions, yet little is known about how various biotic factors such as sex, parasites, and ontogeny influence (singly and via interactions) the stress response of eels. In this study, mixed-effects and linear models were used to quantify the influence of sex, parasitism (Anguillicola crassus), life stage (yellow and silver eels), and silvering stage on the stress response of eels when exposed to a standardized handling stressor. The physiological response of eels to a standardized abiotic stressor (netting confinement in air) was quantified through measurements of blood glucose and plasma cortisol. The relationships between biotic factors and the activity of gill Na+/K+-ATPase was also examined. Analyses revealed that in some instances a biotic factor acted alone while in other cases several factors interacted to influence the stress response. Blood glucose concentrations increased after exposure to the standardized stressor and remained elevated after 4 h. Variation in plasma cortisol concentrations after exposure to the stressor were found to be time dependent, which was exacerbated by life stage and parasitism condition. Males and nonparasitized silver eels had the highest Na+/K+-ATPase activity. Silvering stage was strongly positively correlated with Na+/K+-ATPase activity in female eels. Collectively, these findings confirm that the factors mediating stress responsiveness in fish are complicated and that aspects of inherent biotic variation cannot be ignored.

Detection of ascaridoid nematode parasites in the important marine food-fish Conger myriaster (Brevoort) (Anguilliformes: Congridae) from the Zhoushan Fishery, China.

BACKGROUND: The whitespotted conger Conger myriaster (Brevoort) (Anguilliformes: Congridae) is an extremely marketable food fish, commonly consumed as sashimi or sushi in some Asian countries (i.e. Japan, Korea and China). Conger myriaster is also suspected as being an extremely important source of human anisakidosis. However, there is currently very little information on the levels of infection with ascaridoid nematode parasites in this economically important marine fish. The aims of the present study are to determine the species composition, prevalence and mean intensity of ascaridoid parasites of C. myriaster caught in the Zhoushan Fishery. RESULTS: A total of 1142 third-stage ascaridoid larvae were isolated from 204 C. myriaster. The overall prevalence of infection was 100% (mean intensity 5.6). Nine species of such larvae were accurately identified using integrative taxonomic techniques involving both morphological and genetic data; these included Anisakis pegreffii, A. typica and A. simplex (sensu stricto) × A. pegreffii, Hysterothylacium fabri, H. aduncum, H. sinense, H. amoyense, H. zhoushanense and Raphidascaris lophii. Although high levels of infection and species richness were revealed in C. myriaster, most of the ascaridoid parasites (1135 individuals) were collected from the body cavity and visceral organs of the fish and only seven individuals of A. pegreffii were found in the musculature. CONCLUSIONS: This study represents the first report C. myriaster from the Zhoushan Fishery being heavily infected with third-stage ascaridoid larvae. Among the ascaridoid larvae parasitic in this fish, an important etiological agent of human anisakidosis, A. pegreffii (L3), represents the predominant species. The genus Hysterothylacium has the highest species richness, with H. fabri (L3) being the most prevalent species. This high level of infection of A. pegreffii (L3) in C. myriaster suggests a high risk of anisakidosis or associated allergies for people consuming raw or poorly cooked fish originating from this marine area. These findings provide important basic information on the occurrence and infection parameters of ascaridoid nematodes in this economically important marine fish. They also have significant implications for the prevention and control of human anisakidosis when conger eels from the Zhoushan Fishery are consumed.

Neoascarophis sinensis n. sp. (Nematoda: Cystidicolidae), a parasite of Conger myriaster (Brevoort) (Anguilliformes: Congridae) in Chinese waters.

Neoascarophis sinensis n. sp. collected from the whitespotted conger Conger myriaster (Brevoort) (Anguilliformes: Congridae) in the Yellow and East China Seas, is described using both light and scanning electron microscopy. The new species is characterised mainly by the body size (8.5-10.5 mm in the males, 9.5-14.0 mm in the females), the location of the vulva (near equatorial region of the body), the non-bifurcate deirids, the lengths of the vestibule (40-50 µm in the males, 30-60 µm in the females) and glandular oesophagus (2.5-3.1 mm in the males, 3.1-3.5 mm in the females) and the morphology and length of the spicules (left spicule 400-410 µm, right spicule 130-150 µm). Neoascarophis sinensis n. sp. is the first species of Neoascarophis Machida, 1976 reported from the anguilliform fish and is also the only species of this genus found in the Chinese waters.

Genetic diversity of infective larvae of Gnathostoma spinigerum (Nematoda: Gnathostomatidae) in freshwater swamp eels from Thailand.

Human gnathostomiasis is a food-borne zoonosis caused by a tissue nematode of the genus Gnathostoma. The disease is highly endemic in Asia, including Thailand. The freshwater swamp eel (Monopterus albus), the second intermediate host of the gnathostome nematode, has an important role in transmitting the infection in Thailand. Surveys on the infective larvae of Gnathostoma spinigerum based on morphological features in freshwater swamp eels have been performed continuously and reported in Thailand. However, there is still limited molecular data on intra-species variations of the parasite. In this study, a total of 19 third-stage larvae of morphologically identified G. spinigerum were collected from 437 liver samples of freshwater swamp eels purchased from a large wholesale market in Bangkok, Thailand. Molecular characterization based on mitochondrial cytochrome c oxidase subunit I (COI) sequences was performed to elucidate their genetic variations and phylogenetic relationship. Among the 19 infective larvae recovered from these eels, 16 were sequenced successfully. Phylogenetic analyses inferred from the partial COI gene showed the presence of three distinct COI haplotypes. Our findings confirm the presence of G. spinigerum as the main species in Thailand.

A new species of microphallid (Trematoda: Digenea) infecting a novel host family, the Muraenidae, on the northern Great Barrier Reef, Australia.

A new species of microphallid, Longiductotrema tethepae n. sp., is reported from a muraenid eel Gymnothorax pseudothyrsoideus (Bleeker) on the northern Great Barrier Reef. The new species is described based on adults from Gy. pseudothyrsoideus and metacercariae from a grapsid crab, Grapsus albolineatus Latreille in Milbert, collected from off Lizard Island, Queensland, Australia. The new species is assigned to Longiductotrema Deblock & Heard, 1969 based on morphological characters (presence of a cirrus-sac; a long metraterm, intensively ensheathed by gland-cells; an entirely postcaecal uterus; vitellarium composed of two lateral clusters each of about ten follicles, situated in the testicular and post-testicular areas). Longiductotrema tethepae n. sp. is the third species assigned to the genus, differing from its congeners in having a distinctly larger body dimensions, a smaller pharynx in relation to oral sucker, the anterior limits of the vitelline fields at the level of the testes (vs at the level of the ovary) and in its parasitism of a muraenid fish (vs birds). Additionally, the new species differs from L. floridense Deblock & Heard, 1969 in having a shorter metraterm and from L. scandolensis Deblock & Bartoli, 1988 in having a less elongate forebody in relation to body length, shorter caeca and prepharynx, and slightly larger eggs. Phylogenetic analyses, based on partial 28S rRNA gene sequences, showed that the present species is sister to all other microphallids for which sequence data are available. This is the fourth report of a microphallid from a marine eel, the first from the Muraenidae Rafinesque and the first from a marine fish in the Indo-west Pacific. A summary of all species of the Microphallidae parasitising fish is provided.

Cross-Reactivity Pattern of Asian and American Human Gnathostomiasis in Western Blot Assays Using Crude Antigens Prepared from Gnathostoma spinigerum and Gnathostoma binucleatum Third-Stage Larvae.

Gnathostomiasis is a zoonotic parasitosis endemic in many Asian and some Latin American countries. Most human infections are caused by Gnathostoma spinigerum in Asia and Gnathostoma binucleatum in the Americas, and recently, imported cases have been increasing among travelers returning from endemic regions. Confirmation of the clinical diagnosis relies largely on serologic tests, with a G. spinigerum-antigen-based immunoblot currently being the diagnostic method of choice. However, we repeatedly experienced that sera from patients with clinically suspected American gnathostomiasis gave negative results in this assay. Therefore, we used homologous methods to prepare G. spinigerum- and G. binucleatum-antigen-based immunoblot assays, and evaluated the cross-reactivity of the two assays. The results show incomplete cross-reactivity between the two assays: the G. spinigerum-antigen-based immunoblot apparently only detects Asian gnathostomiasis caused by G. spinigerum, whereas the G. binucleatum-antigen-based immunoblot is apparently capable of detecting American as well as Asian gnathostomiasis.

Inactivation of Anisakis simplex L3 in the flesh of white spotted conger (Conger myriaster) by high hydrostatic pressure and its effect on quality.

Koreans consume much seafood; the country is surrounded on the east, west and south by the sea. Koreans have eaten raw sashimi for a long time. However, a concern in the raw sea food industry is that the parasitic nematode Anisakis simplex L3 occurs naturally in marine fish. Thus, the fishery industry needs a non-thermal processing method. High hydrostatic pressure (HPP) has been demonstrated to be effective. White spotted conger flesh containing 20 live larvae was exposed to different pressures (150 and 200 MPa for 1 and 5 min; 250 and 300 MPa each for 1 min). The viability of A. simplex L3 was significantly (p < 0.05) reduced in the flesh of white spotted conger by the stepwise increase of high pressure and time. The conditions required to eliminate A. simplex L3 were as follows: 200 MPa for 5 min or 300 MPa for 1 min. The flesh of the white spotted conger treated at 300 MPa for 1 min was whiter and yellower than untreated controls or that treated at 200 MPa for 5 min. No significant changes (p > 0.05) in any of the Hunter colour ('L', 'a' and 'b') values were found after HPP at 200 MPa for 5 min. The fresh treated at 300 MPa for 1 min scored < 4.0 (the defect limit of quality) of flavour, texture and overall acceptability in untrained sensory evaluation using a seven-point hedonic scale. However, the flesh treated at 200 MPa for 5 min scored > 5.0 ('like') for all sensory parameters. This study suggested that HPP at 200 MPa for 5 min could potentially be used for the inactivation of A. simplex L3 in raw fishery food products without any concomitant changes in their colour or sensory qualities.

Dollfustrema durum n. sp. and Heterobucephalopsis perardua n. sp. (Digenea: Bucephalidae) from the giant moray eel, Gymnothorax javanicus (Bleeker) (Anguilliformes: Muraenidae), and proposal of the Heterobucephalopsinae n. subfam.

Two new species of bucephalid trematode (Platyhelminthes: Digenea) are described from the giant moray eel, Gymnothorax javanicus (Anguilliformes: Muraenidae), from off Lizard Island, Australia. We used a combined morphological and molecular-based approach targeting the internal transcribed spacer 2 (ITS2) of the ribosomal DNA (rDNA) and the D1-D3 region of the large subunit (28S) of rDNA to circumscribe the species. Dollfustrema durum n. sp. is distinguished from seven congeners in having 5-6 rows of enlarged body spines circling the anterior portion of the rhynchus. From the remaining 10 species, D. durum n. sp. differs in body length, and in having a caecum that terminates posteriorly to the confluent arc formed by the vitelline follicles, gonads predominantly anterior to the pharynx, testes in tandem, an anterior testis positioned posteriorly to the vitelline follicles, and the pre-vitelline field 23-40% of the body length. Heterobucephalopsis perardua n. sp. differs from Heterobucephalopsis gymnothoracis, the type- and only other reported species, in being two to three times smaller. Heterobucephalopsis, currently considered a genus inquirendum, is confirmed as valid and is rediagnosed. Bayesian inference analysis of 28S rDNA sequences representing 28 species from nine genera and four subfamilies of bucephalid, indicates that i) subfamily classifications previously based on morphological characters are broadly robust, ii) the sequence representing H. perardua n. sp. is resolved as distinct, and basal, to sequences representing the Bucephalinae, the Prosorhynchinae, the Paurorhynchinae, and the Dolichoenterinae, iii) the Dolichoenterinae and the Prosorhynchinae are monophyletic sister clades, basal to the Bucephalinae and the Paurorhynchinae, iv) sequences representing Grammatorcynicola, Prosorhynchus, and Dollfustrema are also monophyletic, v) the Bucephalinae is paraphyletic relative to the Paurorhynchinae, and vi) the bucephaline genera Prosorhynchoides, Rhipidocotyle, and Bucephalus are each polyphyletic. The morphological and molecular differences observed among the four previously recognised subfamilies in this study lead us to propose Heterobucephalopsinae n. subfam. to accommodate the genus Heterobucephalopsis.

Description of a new species of Podocotyle Dujardin, 1845 (Digenea: Opecoelidae: Plagioporinae) from the cusk-eel, Luciobrotula corethromycter Cohen, 1964 (Ophidiiformes: Ophidiidae), from the Gulf of Mexico and Caribbean Sea.

Podocotyle bathyhelminthos n. sp. (Opecoelidae: Plagioporinae) is described from the cusk-eel, Luciobrotula corethromycter Cohen, 1964 (Ophidiiformes: Ophidiidae), collected at depths of 622-1,280 m in the northern Gulf of Mexico and Caribbean Sea off Colombia. The new species is distinguished by possessing a combination of the following features: an elongate oval body shape, a sucker width ratio of 1:1.4-1.9, deeply lobed and irregularly-shaped testes, a cirrus-sac that extends just in to the hindbody, a trilobed ovary and vitellarium that extend to the ventral sucker level. Several unique features in P. bathyhelminthos n. sp. were not present in all, or almost all, recognized species of Podocotyle including a conspicuous deep cleft at the posterior end of the worm, a small transverse ridge on the ventral surface immediately anterior to the ventral sucker, uterine loops extending ventral to the caeca and, at times, lateral to the caeca, a thick-walled metraterm extending 1/3 to 1/2 the length of the cirrus-sac and P. bathyhelminthos n. sp. parasitizes a deep water piscine host. Podocotyle etheostomae Aliff, 1973 is declared a nomen nudum. A brief discussion of Podocotyle Dujardin, 1845 in deep waters is presented, and a gastropod, caridean shrimp and cusk-eel are hypothesized as hosts in the life cycle of P. bathyhelminthos n. sp. in the deep sea.

Experimental anisakid infections in mice.

Anisakidosis is a human parasitic disease caused by infections with members of the Anisakidae family. Accidental infection after fish intake affects the gastrointestinal tract as a consequence of mechanical damage caused by migrating larvae. Infections can also trigger allergies, hives, severe asthma or anaphylaxis with angioedema. Although mouse models of intraperitoneal antigenic stimulation exist, enabling immunological studies, few models using gastric introduction of live larvae are available for the study of immunological and gastrointestinal damage in mice. This study was designed to characterize serum reactivity against Anisakis spp. and Contracaecum spp. in Balb/c mice following orogastric inoculation and to assess gastrointestinal damage. These anisakid species were classified at the Universidade Federal Fluminense (UFF) School of Veterinary Medicine and materials for live larval inoculation were developed at the UFF Immunobiology laboratory. Live larvae were inoculated following injection with a NaCl solution. Blood samples were collected and sera screened for immunoglobulin (Ig)E and IgG anti-larva responses to both nematodes, specific for somatic and excretory/secretory antigens, by enzyme-linked immunosorbent assay (ELISA). The means of the optical densities were analysed using analysis of variance (ANOVA) with Tukey's post-hoc test and the general linear model. This analysis identified the presence of anti-IgG seroreactivity to both somatic and excretory/secretory Anisakis antigens in inoculated animals compared with controls (P< 0.001), and no gastric or intestinal damage was observed. These experiments demonstrated that introduction of live Contracaecum spp. into the gastrointestinal tract did not elicit serum sensitization in animals.

Richness and diversity of helminth species in eels from a hypersaline coastal lagoon, Mar Menor, south-east Spain.

The composition and diversity of parasite communities and intestinal components, as well as infra-community structure, were assessed in eels Anguilla anguilla, from Mar Menor, a permanent Mediterranean hypersaline coastal lagoon. Data were used to determine whether this helminth community differs in composition and structure from that of eels in lagoons with lower salinity regimes and higher freshwater inputs. A total prevalence of 93% was detected. Specifically, parasites were identified as Deropristis inflata, Bucephalus anguillae, Contracaecum sp., Anguillicoloides crassus and two plerocercoid larvae belonging to the order Proteocephalidae, the marine species representing 91% of the isolated helminths. In the total community, digenetic trematodes were the dominant group of helminths, and D. inflata, an eel specialist, dominated both the component community and the infra-community. Richness and diversity were low but similar to those reported in other saline lagoons, and maximum species per eel did not exceed four. At the infra-community level, higher abundance than in other brackish or marine Mediterranean environments was detected. The findings provide further evidence of the similarity in composition and structure of helminth communities in eels from various Mediterranean coastal lagoons. Moreover, salinity-dependent specificities are well supported and reflect the life history of individual eels.

Nematode eel parasite found inside acanthocephalan cysts--a "Trojan horse" strategy?

BACKGROUND: The invasive eel parasite Anguillicoloides crassus (syn. Anguillicola crassus) is considered one of the major causes for the decline of the European eel (Anguilla anguilla) panmictic population. It impairs the swim bladder function and reduces swimming performance of its host. The life cycle of this parasite involves different intermediate and paratenic hosts. Despite an efficient immune system of the paratenic fish hosts acting against infections with A. crassus, levels of parasitized eels remain high in European river systems. Recently, the round goby Neogobius melanostomus (Gobiidae) has become dominant in many rivers in Europe and is still spreading at a rapid pace. This highly invasive species might potentially act as an important, so far neglected paratenic fish host for A. crassus. METHODS: Based on own observations and earlier single sightings of A. crassus in N. melanostomus, 60 fresh individuals of N. melanostomus were caught in the Rhine River and examined to assess the infection levels with metazoan parasites, especially A. crassus. Glycerin preparations were used for parasite identification. RESULTS: The parasite most frequently found in N. melanostomus was the acanthocephalan Pomphorhynchus sp. (subadult stage) which occurred mainly encysted in the mesenteries and liver. Every third gobiid (P = 31.7%) was infected by A. crassus larvae (L3) which exclusively occurred inside the acanthocephalan cysts. No intact or degenerated larvae of A. crassus were detected elsewhere in the goby, neither in the body cavity and mesenteries nor in other organs. Affected cysts contained the acanthocephalan larvae and 1-12 (mI =3) living A. crassus larvae. Additionally, encysted larvae of the nematode Raphidascaris acus were detected in the gobies, but only in the body cavity and not inside the acanthocephalan cysts. CONCLUSIONS: Based on our observations, we suggest that A. crassus might actively bypass the immune response of N. melanostomus by invading the cysts of acanthocephalan parasites of the genus Pomphorhynchus using them as "Trojan horses". Providing that eels prey on the highly abundant round goby and that the latter transfers viable infective larvae of A. crassus, the new paratenic host might have a strong impact on the epidemiology of A. crassus.

Morphological and molecular characterization of Cucullanus hainanensis sp. nov. (Ascaridida: Cucullanidae) from Muraenichthys gymnopterus (Bleeker) (Anguilliformes: Ophichthidae) in the South China Sea.

Cucullanus hainanensis sp. nov., collected from Muraenichthys gymnopterus (Bleeker) (Anguilliformes: Ophichthidae) in the South China Sea, was described using both light and scanning electron microscopy. The new species can be readily distinguished from its congeners by the large pseudobuccal capsule, the position of excretory pore and deirids, the length of spicules (0.64-0.76 mm, 5.84-6.67% of body length) and gubernaculum (0.21-0.24 mm), the number and arrangement of caudal papillae and the particular morphology of cloacal region in male. The new species was also characterized using molecular methods by sequencing and analysing the small subunit (18S) and the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA (rDNA). In addition, Cucullanus muraenesocis (Yin et Zhang, 1983) was regarded a homonym of C. muraenesocis Yamaguti, 1961, and a new name, Cucullanus wangi nom. nov. was given to it.

Can differences in life cycle explain differences in invasiveness? - A study on Anguillicola novaezelandiae in the European eel.

Anguillicola crassus is the most invasive species of its genus and it is a successful colonizer of different eel species worldwide. It is so far the only species of the genus Anguillicola whose life cycle has been studied completely. To analyse whether differences in life cycle may explain differences in invasiveness, we infected European eels with Anguillicola novaezelandiae under laboratory conditions. Anguillicola novaezelandiae shows a synchronized development in the European eel. Eggs with second-stage larvae appeared 120 days after infection. No density-dependent effect in parasite development could be found for A. novaezelandiae. The present study shows that the life cycle of A. novaezelandiae differs on final host level compared with A. crassus in ways which result in a less successful invasion of new host species.

Peracreadium akenovae sp. nov. (Trematoda: Opecoelidae) parasitising the highfin moray eel Gymnothorax pseudothyrsoideus (Anguilliformes: Muraenidae) from Moreton Bay, Australia.

A new opecoelid trematode, Peracreadium akenovae sp. nov., is described from the highfin moray eel Gymnothorax pseudothyrsoideus (Bleeker) (Anguilliformes; Muraenidae), collected from Moreton Bay off southeast Queensland, Australia. The new species is distinctive in its body shape, transversely elongate irregular testes, vitelline follicles interrupted at the level of the ventral sucker, and diverticulate excretory vesicle. The Muraenidae is a new host family for Peracreadium Nicoll, 1909. Peracreadium is the seventh opecoelid genus reported from temperate eastern Australian marine fishes and this is its first report from Australian waters.

A new species of Podocotyloides (Digenea: Opecoelidae) from the grey conger eel, Conger esculentus, in the Caribbean Sea.

Podocotyloides brevis n. sp. (Digenea: Opecoelidae) is described from Conger esculentus Poey, 1861, from the Caribbean Sea, off the coast of Mayagüez, Puerto Rico. It can be separated from the other 13 accepted species in the genus by having a shorter cirrus sac, both in length and relative length. Podocotyloides brevis, with a cirrus sac length less than 10% of the body length, most closely resembles Podocotyloides dorabus Lokhande, 1990, which has a cirrus sac that is approximately 16% of the body length. It can be further differentiated from P. dorabus by having ovarian and testicular lengths greater than the oral sucker length. This is the first report of a member of this genus from the Caribbean Sea and from a host species collected as deep as 200 m. A key to the recognized members of Podocotyloides Yamaguti, 1934, is also presented.