Rapid fluctuation in infection levels of Neoheterobothrium hirame (Monogenea) in Japanese flounder Paralichthys olivaceus in the Joban area, Japan

Neoheterobothrium hirame a haematophagous monogenean parasite on Japanese flounder Paralichthys olivaceus occurred at high prevalences (>70%) from 1999 to 2002 but thereafter declined and remained low (<20%) from 2003 to 2005 in the Joban area, Japan. In 2006, N. hirame became abundant again reaching a prevalence of 63%. Generalized linear models explained these rapid fluctuations in infection in relation to inshore water temperature, which affected the reproductive success of the parasite, and also the encounter rate between host and parasite through changes in their respective population densities. Severely anaemic fish were few, 2·4% even in 1999–2002 and 2006, suggesting that the effect of N. hirame infection on the P. olivaceus population was small in this area.     

Impacts of diclidophorid monogenean infections on fisheries in Japan

Many monogeneans are pathogenic to economically important fish in Japan. However no other monogenean is comparable with the diclidophorids, Heterobothrium okamotoi and Neoheterobothrium hirame, on the scale of impacts they inflict on Japanese fisheries. The shared importance of the two monogenean infections lies in their pathogenicity, fecundity and tolerance to chemical treatment. Heterobothrium okamotoi infects the gills and wall of the branchial cavity of the tiger puffer, Takifugu rubripes (Tetraodontidae), which is widely cultured in western Japan. The main presenting signs of infected fish are anaemia and extensive necrosis caused by adult worms. This monogenean deposits long strings of eggs, which reach lengths of almost 3m. Egg entanglement with the mesh of culture nets increases the chance of hatched larvae encountering susceptible fish. The oncomiracidium maintains infectivity for up to 4 days after hatching. Hydrogen peroxide is the only commercially available chemical able to control the infection, but can only kill immature worms on the gills. Neoheterobothrium hirame infects the gills and wall of the buccal cavity of the Japanese flounder, Paralichthys olivaceus (Paralichthyidae). Since the first known occurrence of this monogenean in 1993, the species has been recorded from almost all areas where the host is distributed. Neoheterobothrium hirame has the potential to produce 781 eggs per day at 20 degreeC. In the western Sea of Japan, wild young-of-the-year flounder became infected in early summer, followed by a sharp increase in prevalence in late summer. By late summer, juvenile flounder have nearly disappeared from the area, strongly suggesting that N. hirame is responsible for mortality of young fish. This is in good agreement with the recent decline in the local flounder population. Neoheterobothrium hirame has also been considered the causative agent of anaemia among wild Japanese flounder since the late 1990s.     

Role of cellular response in elimination of the monogenean Neoheterobothrium hirame in Japanese flounder Paralichthys olivaceus

Adult worms of the blood-feeding monogenean parasite Neoheterobothrium hirame, which cause anemia in the Japanese flounder Paralichthys olivaceus, attach to the host fish by embedding their posterior part deeply into the host tissue. To investigate the possibility that cellular responses of the host fish can eliminate N. hirame, flounder were experimentally infected with N. hirame larvae and reared in either fed or starved conditions. Mature parasites were identified on the buccal cavity wall of the fish 33 d post-infection (Day 33). Monocytes/macrophages and granulocytes increased rapidly in the blood and infected sites after the appearance of mature parasites. These cells adhered to the tegument of the parasites. In addition, a few cells with large electron-dense granules (DGCs) were observed in the inflammatory foci. On Day 47, the tegument of some parasites collapsed partially and were phagocytosed by the infiltrated host cells. Some infiltrated cells adhered directly to the inner tissues of the parasites. On Day 54, in the fed fish group, the loss of the tegument led to damage of the parasites' inner tissue by a large number of infiltrated cells. In this group, the elimination of the parasites was noted from Day 47 to 54. These observations probably suggest that the cellular response of the host fish destructed the parasite's posterior part embedded in the tissue, thereby eliminating the parasites. On the other hand, a high mortality was observed in the starved group. The starved fish developed much more severe anemia than the fed fish, and the elimination of the parasites was not observed in this group. The results of the present study suggest that flounder can eliminate N. hirame if they are fed sufficiently.     

Spread of Neoheterobothrium hirame (Monogenea), a serious pest of olive flounder Paralichthys olivaceus, to Korea

Neoheterobothrium hirame is a large, blood-feeding gill-worm infecting the highly prized olive flounder Paralichthys olivaceus in Japan. There is strong evidence that this worm is the primary cause of anaemia, a common and serious condition causing losses among both wild and cultured olive flounders. N. hirame was first detected in Japanese waters less than a decade ago, and its population then proliferated and spread throughout most of Japan, except Hokkaido. In neighbouring Korea, olive flounder is the most important species of cultured marine fish, and production currently exceeds that in Japan. However, until now, there have been no reports of any monogeneans or anaemia among olive flounders in Korea. Our survey conducted in 2000 of 100 cultured individuals from 4 provinces revealed 2 immature specimens of N. hirame: 1 from a land-based pond-culture system in southern Cheju Island (off the SW coast of Korea) and the other from a floating net cage near Yosu (in the mid-S part of the peninsula). The geographic range of this pathogen may have been enlarged as a result of introduction(s) of infected broodstock from Japan, but this seems unlikely. (The raising of this species in hatcheries developed in Korea in 1985, 7 years before the earliest detection of the worm in Japan.) Low numbers of flounders were also clearly anaemic. This, and the current rarity of N. hirame in Korean farms, appears to favour the hypothesis of a more recent, natural dispersal of the worm, during migrations of infected flounder across the narrow and shallow Tsushima and Korea Straits. Regardless of route of entry, we expect this pathogen will have an impact on Korean flounder fisheries equally serious to that being experienced in Japan.     

Behavioral and ecological correlates of foureye butterflyfish, Chaetodon capistratus, (Perciformes: Chaetodontidae) infected with Anilocra chaetodontis (Isopoda: Cymothoidae)

We observed the behavior and ecology of Chaetodon capistratus infected and uninfected with the ectoparasitic isopod Anilocra chaetodontis to assess whether there may be parasite induced alterations in host biology, host defenses against infection, and/or pathology related to infection. We also examined habitat related differences in infection rates. Infected fish had higher rates of interaction with conspecifics and spent more time in low flow environments (which might improve transmission of juvenile parasites to new hosts). Butterfly fish without isopods were chased more frequently by damselfishes, fed more, and had larger territories. Time spent near conspecifics, and fish condition and gonadosomatic index did not vary between infected and uninfected fish. These results suggest that foureye butterfly fish behavior is altered by the isopod parasite in order for the isopods to more easily gain mates or transmit offspring to new hosts.     

Estimates of daily mortality from a neascus trematode in age-0 shortnose sucker (Chasmistes brevirostris) and the potential impact of avian predation

Parasites may be an important component of early life mortality in fishes, but assigning part of total mortality to parasites is difficult. The Chapman-Robson mortality estimator is a robust and potentially valuable way to quantify the added mortality of parasites when age data are available. We used daily age data and the Chapman-Robson catch-curve procedure to estimate daily mortality for 15 years in juvenile age-0 shortnose suckers (Chasmistes brevirostris), and for 6 years, the daily mortality of fish with and without black spot infection, a trematode whose final host is a piscivorous bird. Infected fish always had higher mortality rates than uninfected fish, and for 3 years when those differences were significant, the added daily mortality for infected fish was 3.6–3.7 %. Based on the proportion infected each year, and for durations of 15–50 d, juvenile populations were 18.3–38.6 % lower than they would have been without black spot infections. There were no significant differences in growth between infected and uninfected fish in most years and little indication of a direct metabolic impact of infections. Thus, this added mortality, primarily in July and August, seemed unlikely to be an indirect result of infection and was most likely due to predation. The source of that predation is unknown but the parasite’s final hosts, piscivorous birds, seemed the most obvious candidate for this added mortality.     

An outbreak of VHSV (viral hemorrhagic septicemia virus) infection in farmed Japanese flounder Paralichthys olivaceus in Japan.

A rhabdoviral disease occurred in farmed populations of market sized Japanese flounder (hirame) Paralichthys olivaceus in the Seto Inland Sea of Japan in 1996. The causative agent was identified as viral hemorrhagic septicemia virus (VHSV) based on morphological, immunological, and genetic analyses. Diseased fish that were artificially injected with a representative virus isolate showed the same pathological signs and high mortality as observed in the natural outbreak. This is the first report of an outbreak of VHSV infection in cultured fish in Japan. Clinical signs of diseased fish included dark body coloration, an expanded abdomen due to ascites, congested liver, splenomegaly, and a swollen kidney. Myocardial necrosis was most prominent and accompanied by inflammatory reactions. Necrotic lesions also occurred in the liver, spleen and hematopoietic tissue, and were accompanied by circulatory disturbances due to cardiac failure. Hemorrhagic lesions did not always appear in the lateral musculature. Transmission electron microscopy revealed many rhabdovirus particles and associated inclusion bodies containing nucleocapsids in the necrotized myocardium. The histopathological findings indicated that the necrotizing myocarditis could be considered a pathognomonic sign of VHSV infection in Japanese flounder.     

Experimental horizontal transmission of viral hemorrhagic septicemia virus (VHSV) in Japanese flounder Paralichthys olivaceus

Infection by viral hemorrhagic septicemia virus (VHSV) has recently occurred among wild and farmed Japanese flounder Paralichthys olivaceus in Japan. In the present study, horizontal transmission of VHSV among Japanese flounder was experimentally demonstrated by immersion challenge. Exposure to a flounder isolate (Obama25) of VHSV revealed a dose-response, with higher mortality (81 and 70%) at the 2 higher exposure levels (6.0 and 4.0 log10 TCID50 ml(-1)). In a second experiment, high titers of VHSV were expressed from moribund and dead flounder based on virus detection in holding-tank waters 2 to 3 d prior to death of the fish and 1 d after death. The virus could not be detected in tank waters 2 d after death. Finally, a third cohabitation experiment in small tanks demonstrated horizontal transmission of VHSV from experimentally infected to uninfected fish.     

Reduced growth in wild juvenile sockeye salmon Oncorhynchus nerka infected with sea lice

Daily growth rings were examined in the otoliths of wild juvenile sockeye salmon Oncorhynchus nerka to determine whether infection by ectoparasitic sea lice Caligus clemensi and Lepeophtheirus salmonis was associated with reduced host body growth, an important determinant of survival. Over 98% of the sea lice proved to be C. clemensi and the fish that were highly infected grew more slowly than uninfected individuals. Larger fish also grew faster than smaller fish. Finally, there was evidence of an interaction between body size and infection status, indicating the potential for parasite‐mediated growth divergence.     

Distribution and abundance of the salmonid parasite Parvicapsula minibicornis (Myxozoa) in the Klamath River basin (Oregon-California, U.S.A.)

The distribution and abundance of the myxosporean parasite Parvicapsula minibicornis in the Klamath River mirrored that of Ceratomyxa shasta, with which it shares both its vertebrate and invertebrate host. Assay of fish held at sentinel sites and water samples collected from those sites showed that parasite prevalence was highest below Iron Gate dam, which is the barrier to anadromous salmon passage. Above this barrier parasite levels fluctuated, with the parasite detected in the free-flowing river reaches between reservoirs. This was consistent with infection prevalence in the polychaete host, Manayunkia speciosa, which was greater than 1% only in populations tested below Iron Gate dam. Although a low prevalence of infection was detected in juvenile out-migrant fish in the Trinity River, the tributaries tested did not appear to be a significant source of the parasite to the mainstem despite the presence of large numbers of infected adult salmon that migrate and spawn there. Rainbow trout became infected during sentinel exposure, which expands the host range for P. minibicornis and suggests that wild rainbow trout populations are a reservoir for infection, especially above Iron Gate dam. High parasite prevalence in the lower Klamath River is likely a combined effect of high spore input from heavily infected, spawned adult salmon and the proximity to dense populations of polychaetes.     

Parasitic mites influence fitness components of their host, the land snail Arianta arbustorum

Abstract. Parasites can influence the population dynamics of their hosts by affecting life-history strategies and behavior. The hematophageous mite Riccardoella limacum lives in the lung cavity of terrestrial gastropods. We used correlational and experimental approaches to investigate the influence of parasite infection on the behavior and life-history traits of the simultaneously hermaphroditic land snail Arianta arbustorum , a common host of R. limacum. Naturally infected individuals of A. arbustorum , collected in the wild, showed a decreased activity compared with uninfected snails. The reproductive output, expressed as the number of eggs deposited in a reproductive season, was reduced in mite-infected hosts. However, the hatching success of the eggs laid by parasitized snails was slightly higher than that of uninfected individuals. We also examined winter survival in 361 adults of A. arbustorum collected from four natural populations. The prevalence of mite infection ranged from 44.8% to 70.1% in three populations (snails in the fourth population were not infected). Winter survival was reduced in infected snails in two out of three populations. Furthermore, experimentally infected snails from an uninfected population showed a reduced winter survival compared with control snails. Our results indicate that parasite pressure imposed by members of R. limacum may influence life history in A. arbustorum.     

Effects of host migration, diversity and aquaculture on sea lice threats to Pacific salmon populations

Animal migrations can affect disease dynamics. One consequence of migration common to marine fish and invertebrates is migratory allopatry-a period of spatial separation between adult and juvenile hosts, which is caused by host migration and which prevents parasite transmission from adult to juvenile hosts. We studied this characteristic for sea lice (Lepeophtheirus salmonis and Caligus clemensi) and pink salmon (Oncorhynchus gorbuscha) from one of the Canada's largest salmon stocks. Migratory allopatry protects juvenile salmon from L. salmonis for two to three months of early marine life (2-3% prevalence). In contrast, host diversity facilitates access for C. clemensi to juvenile salmon (8-20% prevalence) but infections appear ephemeral. Aquaculture can augment host abundance and diversity and increase parasite exposure of wild juvenile fish. An empirically parametrized model shows high sensitivity of salmon populations to increased L. salmonis exposure, predicting population collapse at one to five motile L. salmonis per juvenile pink salmon. These results characterize parasite threats of salmon aquaculture to wild salmon populations and show how host migration and diversity are important factors affecting parasite transmission in the oceans.     

Effects of a muscle‐infecting parasitic nematode on the locomotor performance of their fish host

The southern flounder Paralichthys lethostigma, host to the nematode Philometroides paralichthydis that is embedded in place of the inclinator muscles of the dorsal and anal fin elements, is hypothesized to impair two aspects of locomotor performance (swimming and burying capacity). Peak swimming acceleration and both measures of burying performance did not differ between infected and uninfected fish, whereas swimming velocity of infected fish was significantly lower than that of uninfected fish. Smaller infected fish swam at significantly slower speeds than smaller uninfected fish, whereas there was no difference among larger fish. Neither the location nor the number of worms affected either swimming or burying performance. The decrease in swimming velocity observed in smaller infected fish may be sufficient in rendering them more vulnerable to predation and environmental stressors.     

Density-dependent predation and implications for stock enhancement with Japanese flounder

In laboratory predation trials kinsen-gani crab Matuta lunaris rarely preyed on hatchery-reared (HR) Japanese flounder Paralichthys olivaceus , even at extremely elevated prey densities ( c . 90 fish m⁻²), but fed readily on sacrificed Japanese founder. This suggests that under similar conditions in the wild predator-prey interactions between kinsen-gani and juvenile Japanese flounder are likely to be weak. Larger Japanese flounder readily cannibalized smaller conspecifics, suggesting that within-release group cannibalism may contribute considerably to post-release mortality of HR Japanese flounder when sufficient size variation exists within groups of released fish. Kinsen-gani presence had no effect on within-release group cannibalism rates of large Japanese flounder on small Japanese flounder. Within-release group cannibalism rates were significantly affected by densities of smaller (prey) fish, implying that the number of fish released (and thus post-release densities) in stock enhancement programmes may significantly affect post-release mortality rates. The probability of within-release group cannibalism increased rapidly as the cannibal-to-prey size ratio exceeded 1.6, suggesting that minimizing size variation of released HR individuals will reduce post-release cannibalism rates. These results have implications for Japanese flounder stock enhancement release strategies, and may be applicable to other finfish stock enhancement programmes.     

Parasite-induced suppression of aggregation under predation risk in a freshwater amphipod: Sociality of infected amphipods

Recent findings suggest that grouping with conspecifics is part of the behavioural defences developed by amphipod crustaceans to face predation risk by fish. Amphipods commonly serve as intermediate hosts for trophically transmitted parasites. These parasites are known for their ability to alter intermediate host phenotype in a way that promotes predation by definitive hosts, where they reproduce. If aggregation in amphipods dilutes the risk to be preyed on by fish, then it may dilute the probability of transmission for the parasite using fish as definitive hosts. Using experimental infections, we tested whether infection with the fish acanthocephalan Pomphorhynchus laevis alters attraction to conspecifics in the amphipod intermediate host Gammarus pulex. We also measured G. pulex's activity and reaction to light to detect potential links between changes in aggregation and changes in other behaviours. The attraction to conspecifics in the presence of predator cue, a behaviour found in uninfected gammarids, was cancelled by the infection, while phototaxis was reversed and activity unchanged. We found no correlation between the three behaviours in infected amphipods, while activity and aggregation were negatively correlated in uninfected individuals after the detection of predation cue. The physiological causes and the adaptive value of aggregation suppression are discussed in the context of a multidimensional manipulation.     

Effects of diet and<Emphasis Type="Italic"> Echinostoma revolutum</Emphasis> infection on energy allocation patterns in juvenile<Emphasis Type="Italic"> Lymnaea elodes</Emphasis> snails

Resource allocation strategies may be influenced by both biotic and abiotic factors. The purpose of this study was to investigate the effects of both parasitism and diet quality on the growth, reproduction, and survival of the pond snail, Lymnaea elodes. In addition, we assessed parasite growth and reproduction. High-protein (high diet) or low-protein diets (low diet) were fed to juvenile L. elodes snails that were either exposed or sham-exposed to the castrating trematode, Echinostoma revolutum. Host growth was assessed weekly; reproduction and survival were recorded every 2-3 days. We estimated parasite development as the time to parasite release from the host (patency), and parasite reproduction as the number of larvae shed from infected snails at two time points. Diet and infection status had significant effects on snail growth. Infected snails produced few eggs and tended to grow to larger sizes than uninfected snails regardless of diet. In contrast, exposed-uninfected individuals displayed diet-dependent patterns of growth and reproduction. On the high-protein diet, uninfected and exposed-uninfected snails exhibited similar patterns of growth and reproduction, whereas in the low-diet treatment, exposed-uninfected snails exhibited reduced growth and delayed reproduction relative to uninfected individuals. Survival differed among treatments in the latter stages of the study with infected snails exhibiting reduced survival relative to snails from other treatments. Moreover, infected low-diet snails exhibited lower survival than infected high-diet snails. Parasite development and reproduction did not appear to be directly influenced by the quality of host diet. Results from this study suggest that energy allocation patterns are context-dependent in juvenile snails, influenced by parasite exposure and diet quality. Furthermore, parasite reproduction appears to depend more on host size than on the quality of host diet.     

Parasite-induced alteration of plastic response to predation threat: increased refuge use but lower food intake in Gammarus pulex infected with the acanothocephalan Pomphorhynchus laevis

Larvae of many trophically-transmitted parasites alter the behaviour of their intermediate host in ways that increase their probability of transmission to the next host in their life cycle. Before reaching a stage that is infective to the next host, parasite larvae may develop through several larval stages in the intermediate host that are not infective to the definitive host. Early predation at these stages results in parasite death, and it has recently been shown that non-infective larvae of some helminths decrease such risk by enhancing the anti-predator defences of the host, including decreased activity and increased sheltering. However, these behavioural changes may divert infected hosts from an optimal balance between survival and foraging (either seeking food or a mate). In this study, this hypothesis was tested using the intermediate host of the acanthocephalan parasite Pomphorhynchus laevis, the freshwater amphipod Gammarus pulex. We compared activity, refuge use, food foraging and food intake of hosts experimentally infected with the non-infective stage (acanthella), with that of uninfected gammarids. Behavioural assays were conducted in four situations varying in predation risk and in food accessibility. Acanthella-infected amphipods showed an increase in refuge use and a general reduction in activity and food intake. There was no effect of parasite intensity on these traits. Uninfected individuals showed plastic responses to water-borne cues from fish by adjusting refuge use, activity and food intake. They also foraged more when the food was placed outside the refuge. At the intra-individual level, refuge use and food intake were positively correlated in infected gammarids only. Overall, our findings suggest that uninfected gammarids exhibit risk-sensitive behaviour including increased food intake under predation risk, whereas gammarids infected with the non-infective larvae of P. laevis exhibit a lower motivation to feed, irrespective of predation risk and food accessibility.     

Spatiotemporal and gender‐specific parasitism in two species of gobiid fish

Parasitism is considered a major selective force in natural host populations. Infections can decrease host condition and vigour, and potentially influence, for example, host population dynamics and behavior such as mate choice. We studied parasite infections of two common marine fish species, the sand goby (Pomatoschistus minutus) and the common goby (Pomatoschistus microps), in the brackish water Northern Baltic Sea. We were particularly interested in the occurrence of parasite taxa located in central sensory organs, such as eyes, potentially affecting fish behavior and mate choice. We found that both fish species harbored parasite communities dominated by taxa transmitted to fish through aquatic invertebrates. Infections also showed significant spatiotemporal variation. Trematodes in the eyes were very few in some locations, but infection levels were higher among females than males, suggesting differences in exposure or resistance between the sexes. To test between these hypotheses, we experimentally exposed male and female sand gobies to infection with the eye fluke Diplostomum pseudospathaceum. These trials showed that the fish became readily infected and females had higher parasite numbers, supporting higher susceptibility of females. Eye fluke infections also caused high cataract intensities among the fish in the wild. Our results demonstrate the potential of these parasites to influence host condition and visual abilities, which may have significant implications for survival and mate choice in goby populations.     

Field evaluation by RNA/DNA ratios on post-release nutritional status of released and wild Japanese flounder Paralichthys olivaceus juveniles

A large-scale juvenile Japanese flounder (Paralichthys olivaceus) release-recapture experiment was undertaken to find the optimal release season by evaluating the nutritional status of released fish at different seasons during which food abundance was significantly different. Forty thousand fish were released at depths of 1.5 m for early-release (May 29, 1997) and 2 m for late-release (July 2, 1997) (42.1±3.5 and 40.9±4.2 mm body length, respectively) in an experimental field, Wakasa Bay, the Sea of Japan. Samples were taken, after the releases, at Wada beach at intervals of 1, 2, 3, 6, 10, 16 and 30 days after release (DAR), including pre-surveys before each release. Released fish recaptured from the two different release groups totaled 764; 467 from the early-release group (ER) and 297 from the late-release group (LR). A total of 1956 wild flounder juveniles were simultaneously collected (1041 ER, 915 LR). ER fish were subject to higher food availability and were exposed to less pressure from predation by smaller wild juvenile flounder. RNA/DNA ratios in ER juveniles were significantly higher than those of LR fish during all samples. Especially, RNA/DNA ratios in ER juveniles were higher than in wild juveniles from 3 to 50 DAR. In the LR group, the nutritional status of juveniles was relatively low in shallower water. These findings corresponded well with feeding incidence examined by coworkers. Mass release of hatchery-reared juveniles apparently reduced RNA/DNA ratio of the wild juveniles right after releasing. The present study showed that earlier release of hatchery-reared juvenile Japanese flounder with higher RNA/DNA ratio could increase the possibilities of survival right after release in the nursery ground, and that RNA/DNA ratio appeared to be a good tool in evaluating nutritional status of released juveniles as well as wild juveniles in Japanese flounder.     

Evolutional conservation of molecular structure and antiviral function of a viral RNA receptor, LGP2, in Japanese flounder, Paralichthys olivaceus

LGP2 is an important intracellular receptor that recognizes viral RNAs in innate immunity. To understand the mechanism of viral RNA recognition, we cloned an LGP2 cDNA and gene in Japanese flounder (Paralichthys olivaceus). Viral hemorrhagic septicemia virus-induced expressions of LGP2 mRNA were evaluated in vivo and in vitro by quantitative real-time PCR (Q-PCR) using primers based on the clone sequences. The expression of LGP2 mRNA in the kidney dramatically increased at 3 d postinfection. The expression of LGP2 mRNA also increased in the head kidney leukocytes stimulated with artificial dsRNA (polyinosin-polycytidylic acid) in vitro. To evaluate the antiviral activity of the flounder LGP2, three expression constructs containing pcDNA4-LGP2 (full-length), pcDNA4-LGP2ΔRD (regulatory domain deleted), and pcDNA4-Empty (as a negative control) were transfected into the hirame (flounder) natural embryo (hirame natural embryo) cell line. Forty-eight hours after transfection, the transfected cells were infected with ssRNA viruses, viral hemorrhagic septicemia virus, or hirame rhabdovirus. The cytopathic effects of the viruses were delayed by the overexpression of Japanese flounder LGP2. The Q-PCR demonstrated that mRNA expression levels of type I IFN and IFN-inducible genes (Mx and ISG15) in the hirame natural embryo cells overexpressing LGP2 were increased by polyinosin-polycytidylic acid and viral infections. These results suggest that Japanese flounder LGP2 plays an important role in the recognition of both viral ssRNA and dsRNA to induce the antiviral activity by the production of IFN-stimulated proteins.