Infection status of the sea eel (Astroconger myriaster) purchased from the Noryangjin fish market with anisakid larvae]

Although the sea eel (Astroconger myriaster) is suspected as one of the most important fish host for human anisakiasis in Korea, no report has been made on the infection status of the sea eel with anisakid larvae. In the present study, 26 sea eels (Astroconger myriaster) were purchased from the Noryangjin fish market in Seoul, and anisakid larvae were collected from their viscera, muscle, head and skin. The collected larvae were classified by their morphological types. A total of 1,351 anisakid larvae were collected from 15 of 26 fish examined. Among them, 1,269 were recovered from the viscera, 66 from the muscle, and 16 from the head and skin. Morphologically, most of the anisakids were classified into 6 known larval types, Anisakis type I (564 larvae) of Berland (1961), Contracaecum type A (409) and type D (5) of Koyama et al. (1969), Contracaecum type C' (83) and type D' (117) of Chai et al. (1986), and Contracaecum type V (1) of Yamaguti (1935). Remaining 172 specimens were new in the available literature, hence, designated as Contracaecum type A' (new type). The present results revealed that the sea eels caught in the Korean waters are heavily infected with anisakid larvae, not only in their viscera but also in the muscle, and Anisakis type I was the most common among the 7 larval types.     

Larval Anisakid Infections in Marine Fish from Three Sea Areas of the Republic of Korea

The present study was performed to determine the infection status of anisakid larvae in marine fish collected from 3 sea areas of the Republic of Korea. Total 86 marine fish (8 species) collected from the East Sea (Goseong-gun, Gangwon-do), 171 fish (10 species) from the South Sea (Sacheon-si, Gyeongsangnam-do), and 92 fish (7 species) from the Yellow Sea (Incheon Metropolitan City) were examined by both naked eyes and artificial digestion method. Among the total of 349 fish examined, 213 (61.0%) were infected with 8 species of anisakid larvae, i.e., Anisakis simplex, 6 types of Contracaecum spp., and Raphidascaris sp., and the mean larval density was 13.8 per infected fish. Anisakid larvae were detected in 45 fish (52.3%) from the East Sea, 131 fish (76.6%) from the South Sea, and 37 fish (40.2%) from the Yellow Sea. The average numbers of larvae detected were 4.0, 16.6, and 15.9, respectively. Anisakis simplex larvae were detected in 149 fish (42.7%), and the mean larval density was 9.0 per infected fish. They were found in 26 fish (30.2%) collected from the East Sea, 96 fish (56.1%) from the South Sea, and 27 fish (29.3%) from the Yellow Sea. The average numbers of larvae detected were 2.9, 10.3, and 10.5, respectively. Conclusively, the present study suggests that the infection rate and density of anisakid larvae are more or less higher in the fish from the South Sea than those from the East Sea or the Yellow Sea.     

Four cases of gastric submucosal mass suspected as anisakiasis

Anisakiasis is a parasitic disease caused by ingestion of raw fish infected with anisakid larvae. Endoscopic changing patterns of submucosal lesions in chronic gastric anisakiasis have not been known yet. Here we report 4 cases of suspected gastric anisakiasis which were improved during follow-up periods without surgical treatment. The patients presented with abdominal pain, nausea and vomiting after consuming raw marine fish, and visited our gastroenterology outpatient department. Their endoscopic findings showed firm and yellowish submucosal masses accompanied with eccentric erosions. Histologic findings showed severe eosinophilic infiltrations. In blood tests, peripheral eosinophil counts and total IgE levels were elevated. We believed that all cases were caused by larval anisakid infections. The submucosal mass lesions disappeared during the follow-up periods of 2 to 4 mo.     

'Cebiche'--a potential source of human anisakiasis in Mexico?

Five fish species used for preparation of a popular dish (cebiche) made with raw fish flesh in Mexico were obtained from five localities of the coast of Yucatan. Lutjanus synagris, Gerres cinereus, Sphyraena barracuda, Epinephelus morio and Haemulon plumieriwere examined for the presence of larvae of anisakid nematodes, causative agents of human anisakiasis. The nematode Pseudoterranova sp. was found in E. morio and S. barracuda with a total prevalence of 83% and 6.5 +/- 6.2 worms per fish for E. morio, and a prevalence of 33% and 10.2 +/- 30.0 worms per fish for S. barracuda. Contracaecumsp. was found to infect G. cinereus with a prevalence of 57% and 7.6 +/- 11.4 worms per fish. The relatively high prevalence of Pseudoterranova sp. indicates that this parasite is a potential causal agent of anisakiasis on the coast of Yucatan. Although all larvae were found only in the mesentery of the fish host, their importance as a potential source of human infection cannot be excluded as larval migration to the muscles in dead fish is possible.     

Anisakis pegreffii: a quantitative fluorescence PCR assay for detection in situ

To facilitate improved diagnosis and detection of the third stage larva (L3) of Anisakis pegreffii from the Minnan-Taiwan bank fishing ground in Taiwan Strait, a real-time PCR method for the detection in situ and differentiation was developed to amplify a region of the second internal transcribed spacer (ITS-2) of this parasite. The real-time PCR assay was capable of detecting 1/3 of a single L3 in 30 mg of marine fish tissue, and also exhibited a high level of specificity for A. pegreffii, no fluorescence signals were observed in other five major larval anisakid species found in commercial marine fishes caught in this fishing ground.     

Insight into the short-finned squid Illex coindetii (Cephalopoda: Ommastrephidae) feeding ecology: is there a link between helminth parasites and food composition?

Squids are especially frequent as paratenic hosts of helminth parasites, particularly to those that have elasmobranchs and mammals as final hosts. Among those parasite species, anisakid nematode larvae and cestode plerocercoids are most effectively transferred through the trophic chain by oegopsid squids. A total of 439 short-finned squids, Illex coindetii (245 males, 190 females and 4 unsexed) were sampled in the central part of the eastern Adriatic Sea in order to assess their helminth component community and parasite dynamics with respect to host sex, maturity, seasonality, and feeding behavior. Two larval helminths were isolated, i.e., larvae of Anisakis pegreffii, characterized by molecular tools at the species level, and plerocercoids of Phyllobothrium sp., with prevalences of 30.5% and 2.3%, respectively. Highly significant seasonal variation in diet consumption, congruent with seasonal variation in anisakid intensity, was observed, underlining the tight role of squid prey in the trophic transmission of parasite. Likewise, the highest helminth prevalence and intensity of infection was recorded in autumn, when the fish prey, mostly Maurolicus muelleri, comprised the greatest proportion of diet. This helped to assign the Adriatic broadtail shortfin squid not as a first, but as a second, paratenic host for the anisakid, unlike as suggested previously. The presence of larval A. pegreffii confirms its previously reported zoogeographical distribution in the Mediterranean and Adriatic Seas. The presence of 2 helminths in I. coindetii describes the feeding patterns of the squid, as well as clearly defined and coevolved predator-prey relationships.     

Parasites as biological tags in population studies of demersal and pelagic fish species

Among the different techniques applied in a holistic approach for fish stock identification, the use of parasites as "biological tags" is becoming increasingly important. In this presentation, our recent studies on the use of some parasite species, identified by genetic markers, and the parasite/fauna composition, in stock identification of demersal (Merluccius merluccius), small pelagic (Trachurus trachurus), and large pelagic fish species (Xiphias gladius) are reviewed. Different species of Anisakis and Hysterothylacium were genetically identified by the application of genetic (allozyme) markers. Statistically significant differences in the spatial distribution of distinct species of Anisakis were found in the fish considered. As to the species of Hysterothylacium genetically detected, different relative proportions were detected in several Mediterranean and Atlantic samples of swordfish (X. gladius). This study demonstrates the potential value of these anisakid nematodes, at both larval and adult stages, as "biological tags" for these fish species in European waters.     

Multiple primer PCR for the identification of anisakid nematodes from Taiwan Strait

There were six major larval anisakid species found in commercial marine fishes caught in the Minnan fishing ground in the Taiwan Strait: Anisakis physeteris, Anisakis pegreffii, Raphidascaris trichiuri, Contracaecum aduncum, Contracaecum muraenesoxi, Contracaecum sp. For rapid identification of the parasite species above, a single and a multiple primer PCR (multiplex PCR) method, using specific primers based on aligned sequences of the internal transcribed spacer ITS-1, 5.8S, and ITS-2 of nuclear ribosomal DNA, were jointly used for the rapid identification of these anisakid larvae. The primers yielded distinct PCR products for each of the anisakid nematodes, providing rapid and accurate tools for identifying anisakid nematodes with distinct geographical distribution.     

Anisakid nematodes from stranded pygmy sperm whales, Kogia breviceps (Kogiidae), in three localities of the Yucatan Peninsula, Mexico

The present paper reports the presence of 3 adult and juvenile anisakid nematode species: Anisakis simplex, A. brevispiculata, and Pseudoterranova ceticola, which were recovered from the digestive tract of stranded pygmy sperm whales (Kogia breviceps) from 3 localities along the coast of the Yucatan Peninsula. The presence of these anisakid adult nematodes suggests that larval stages may occur in cephalopods or fishes used for human consumption, which represents a potential danger to public health. The occurrence of the 3 anisakid species in coastal waters of the Yucatan Peninsula represents new geographical records for Mexico.     

Intestinal anisakiasis in Italy: case report

A case of intestinal anisakiasis caused by Anisakis sp. larva type I in a woman from Italy who consumed raw marinated anchovies, is reported. The diagnosis was based on the morphological features characteristic of anisakid larval stages, which were readily recognized in a large granuloma removed after emergency surgical treatment.     

The histopathology of Scomber japonicus infection by Nematobothrium scombri (Trematoda:Didymozoidae) and of larval anisakid nematode infections in the liver of Pagrus pagrus

The histopathology induced by Nematobothrium scombri (Trematoda) in Scomber japonicus and of larval anisakid nematodes in Pagrus pagrus is described. Nematodes larvae occurred within the liver capsule and N. scombri occurred within nodules in the opercula. The fishes were collected off the coast of Rio de Janeiro State.     

Larval anisakids parasitizing the blue whiting, Micromesistius poutassou, from Motril Bay in the Mediterranean region of southern Spain

A total of 301 blue whiting, Micromesistius poutassou Risso, 1826, ranging in length from 17 to 28 cm, from Motril Bay (Mediterranean coast, south Spain) were examined for anisakid nematodes, as these fish are common items in the Spanish Mediterranean diet. Three anisakid species were morphologically identified with a total prevalence of 10.63%. Anisakis simplex s.l. Rudolphi, 1809 had a prevalence value of 6.65%, compared with 2.66% for A. physeteris Baylis, 1923 and 2.33% for Hysterothylacium aduncum Rudolphi, 1802. Variations in prevalence values with season and host size are discussed. Allozyme markers (leucine aminopeptidase-1) were used to identify anisakid nematodes assigned to the A. simplex complex and all examined larvae were found to correspond genetically to A. pegreffii Nascetti et al., 1986.     

Infection status of larval anisakids in Astroconger myriaster collected from the Southern Sea near Pusan]

A study was presented on the anisakid larvae in Astroconger myriaster which were caught at the Southern Sea and saled at Pusan area. The nematodes were morphologically classified into Anisakis sp. and Contracaecum sp. after fixation in 70% ethanol and clearing in lactophenol. Total of 1,768 larval anisakids were collected from 259 (67.8% positive rate) out of 382 examined A. myriaster. Total 642 larvae of Anisakis sp. were obtained from 94 (24.6%) and 1,126 Contracaecum sp. were recovered from 165 (43.1%) fish. The average number of worms per infected fish was 4.6. The infection rate increased according to the length of fish, and all of the fish over 71 cm were found infected. The numbers (proportions) of recovered worms by the organs were 1,440 (80.5%) in the omentum, 166 (9.4%) in the intestine, 107 (6.0%) in the stomach, 32 (1.8%) in the skin, 18 (1.0%) in the muscle, 13 (0.7%) in the liver, and 2 (0.1%) in the head. The larvae in the muscle may infect the humans who are enjoying raw sliced meat of the fish.     

Body size,trophic level,and the use of fish as transmission routes by parasites

Within food webs, trophically transmitted helminth parasites use predator–prey links for their own transfer from intermediate prey hosts, in which they occur as larval or juvenile stages, to predatory definitive hosts, in which they reach maturity. In large taxa that can be used as intermediate and/or definitive hosts, such as fish, a host species’ position within a trophic network should determine whether its parasite fauna consists mostly of adult or larval helminths, since vulnerability to predation determines an animal’s role in predator–prey links. Using a large database on the helminth parasites of 303 fish species, we tested whether the proportion of parasite species in a host that occur as larval or juvenile stages is best explained by their trophic level or by their body size. Independent of fish phylogeny or habitat, only fish body length emerged as a significant predictor of the proportion of parasites in a host that occur as larval stages from our multivariate analyses. On average, the proportion of larval helminth taxa in fish shorter than 20 cm was twice as high as that for fish over 100 cm in length. This is consistent with the prediction that small fishes, being more vulnerable to predation, make better hosts for larval parasites. However, trophic level and body length are strongly correlated among fish species, and they may have separate though confounded effects on the parasite fauna exploiting a given species. Helminths show varying levels of host specificity toward their intermediate host when the latter is the downstream host involved in trophic transmission toward an upstream definitive host. Given this broad physiological compatibility of many helminths with fish hosts, our results indicate that fish body length, as a proxy for vulnerability to predators, is a better predictor of their use by helminth larvae than their trophic level based on diet content.     

The role of sexual and asexual reproduction in structuring high latitude populations of the reef coral Pocillopora damicornis

The genotypic composition of populations of the asexually viviparous coral Pocillopora damicornis varies in a manner that challenges classical models of the roles of sexual and asexual reproduction. On the geographically isolated Hawaiian reefs and high latitude reefs in Western Australia, P. damicornis populations are highly clonal although it has been argued that sexual reproduction via broadcast spawning generates widely dispersed colonists. In contrast, on eastern Australia's tropical Great Barrier Reef populations show little evidence of clonality. Here, we compare the genotypic diversity of adult and juvenile colonies of P. damicornis at seven sites on eastern Australia's high latitude Lord Howe Island reefs to determine if levels of clonality vary with habitat heterogeneity and age of colonies (as predicted by theory) or alternatively if clonality is again always high as for other isolated reef systems. We found 55-100% of the genotypic diversity expected for random mating at all seven sites and little evidence of asexual recruitment irrespective of habitat heterogeneity (sheltered versus wave exposed) or colony age. We found reduced levels of genetic diversity compared with tropical reefs (2.75 versus 4 alleles/locus), which supports earlier findings that Lord Howe Island is an isolated reef system. Furthermore, heterozygote deficits coupled with significant genetic subdivision among sites (FST=0.102+/-0.03) is typical of populations that have limited larval connections and are inbred. We conclude that the genetic structure of P. damicornis at Lord Howe Island reflects populations that are maintained through localised recruitment of sexually produced larvae.     

Conflict between antipredator and antiparasite behaviour in larval damselflies

 Larval damselflies resist infestation by parasitic larval mites by exhibiting behaviours such as grooming, crawling, swimming, and striking at host-seeking mites. Larval damselflies are known to increase time spent in these behaviours in the presence of mites but reduce time spent in these behaviours in the presence of fish predators. The presence of both fish and larval mites presents an obvious conflict: a larval damselfly may actively avoid parasitism by mites, thus increasing its risk of predation, or it may reduce its activity when fish are present, thus increasing its risk of parasitism. We analysed the behaviour of larval Ischnura verticalis in an experiment where we crossed presence and absence of fish with presence and absence of larval mites. Presence of mites induced a large increase in activity of larval I. verticalis but fish had no effect and there were no interpretable interactions between effects of mites and fish. Subsequent experiments indicated that larval I. verticalis in the presence of both mites and fish were more likely to be attacked and killed by fish than those exposed only to fish. The high activity level of I. verticalis larvae in the presence of both fish and mites may suggest that costs of parasitism are high, or that under field conditions it is rare for larvae to be in the immediate presence of both fish predators and potentially parasitic mites. Received: 28 March 1996 / Accepted: 6 September 1996     

Further investigation on nuclear transplantation in different orders of teleost: the combination of the nucleus of Tilapia (Oreochromis nilotica) and the cytoplasm of Loach (Paramisgurnus dabryanus).

The nucleus of a blastula cell from Tilapia (Oreochromis nilotica, family Cichlidae, order Perciformes) was transplanted into an enucleated egg of Loach (Paramisgurnus dabryanus, family Cobitidae, order Cypriniformes). From among 3747 nucleo-cytoplasmic hybrid (NCH) eggs two NCH larval fish (0.05%) were obtained; one died on the 6th day and the other died on the 12th day after the operation. Morphological examinations showed that both NCH larval fish had developed normally with an opened mouth except they could not take food after complete utilization of their egg yolk on the 5th day of development. The possible mechanisms for obtaining such inter-order NCH larval fish are discussed. This is the first report indicating that inter-order NCH larval fish can be obtained in spite of their evolutionary divergence.     

Relative larval loss among females during dispersal of Lake Sturgeon (<Emphasis Type="Italic">Acipenser fulvescens</Emphasis>)

Mortality that occurs during larval dispersal as a consequence of environmental, maternal, and genetic effects and their interactions can affect annual recruitment in fish populations. We studied larval lake sturgeon (Acipenser fulvescens) drift for two consecutive nights to examine whether larvae from different females exposed to the same environmental conditions during dispersal differed in relative levels of mortality. We estimated proportional contributions of females to larval collections and relative larval loss among females as larvae dispersed downstream between two sampling sites based on genetically determined parentage. Larval collections were composed of unequal proportions of offspring from different females that spawned at upstream and downstream locations (~0.8 km apart). Hourly dispersal patterns of larvae produced from females spawning at both locations were similar, with the largest number of larvae observed during 22:00–23:00 h. Estimated relative larval loss did not differ significantly among females as larvae were sampled at two sites approximately 0.15 and 1.5 km from the last section downstream of spawning locations. High inter- and intra-female variation in larval contributions and relative larval loss between nights may be a common feature of lake sturgeon and other migratory fish species, and likely is a source of inter-annual and intra-annual variation in fish recruitment.     

Ascorbate concentration in fish ontogeny

The ontogenetic trend of ascorbate has been quantified in three freshwater fishes: roach ( Rutilus rutilus ), whitefish ( Coregonus lavaretus ) and Arctic charr ( Salvelinus alpinus ). Total ascorbate (reduced and oxidized) declined from 150 to μg g⁻¹ as newly hatched larvae grew to become several-months-old juveniles. Declining total ascorbate with increasing size of metamorphosing fish could not be reversed by feeding on brine shrimp, Artemia salina nauplii, zooplanktonic food containing > 74μg g⁻¹ total ascorbate. The proportion of reduced ascorbate in total ascorbate increases with fish size/age. The physiological mechanism of the changes in transferable ascorbate forms remains unknown, but high dehydroascorbate concentrations suggest high vulnerability of larval fish to oxidation stress. This is the first report on quantity of vitamin C retained in actively-feeding larval and juvenile fish. The efficiency of ascorbate transfer from zooplankters to larval fish amounted to 5–20%. The ecological significance of larval fish feeding on various zooplankters and/or phytoplankton may reflect a trend toward maximum transfer of this vitamin in freshwater food webs.