• Fisheries and Aquatic Sciences
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• v.8 no.3
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• pp.185-187
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• 2005

We tested the in vivo ability of a DNA chip to detect virus-specific genes from virus-infected olive flounder Paralichthys olivaceus and rainbow trout Oncorhynchus mykiss. Target cDNA was obtained from total RNA of virus infected cell lines by reverse transcription (RT) and was labeled with fluorescent dye (Cy5-dUTP). The results show the successful detection of infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicaemia virus (VHSV) genes in the virus-infected fishes.

• Journal of fish pathology
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• v.5 no.1
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• pp.1-7
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• 1992

CHSE(Chinook Salmon Embryo)-214 fish cell lines was cultured in Eagle's minimal medium (MEM) supplemented with 10% fetal bovine serum and 2mM-glutamin. Optimum growth temperature of CHSE-214 cell line was $20^{\circ}C$. Infectious pancreatic necrosis virus(IPNV) was successfuly multiplied and showed the cytopathic effect in CHSE-214 cell line. Infection symptom of IPNV was observed with inverted phase contrast microscopy. At 6h-12hrs post-infection, the cells infected with IPNV were similer to normal cells. At 18-24hrs post-infection, the cells were somewhat round form and a little swollen form than normal cells. At 30hs post-infection, the cells were becoming more abnormal cells. At 48-68 post-infection, the infected cells were lysed and showed the severe cytopathic effect.

• Journal of fish pathology
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• v.6 no.2
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• pp.191-195
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• 1993

With the rapid progress in seed production techniques, aquaculture production of economically important species of marine fish has been accelerated in Japan. Howecer, mass mortalities due to viral infections as well as other microbial infections have often occurred during the seed production and grow-out stages. Among these diseases, four viral diseases have been known in cultured Japanese flounder (Paralichthys olivaceus) since around 1980. In this paper, viral diseases of cultured flounder in Japan are briefly reviewed, with special attention to two viral diseases. viral epidermal hyperplasia and rhabdovirus infection which are relatively important because of their frequent occurrence. Viral epidermal hyperplasia is characterized by fin opacity and associated with high mortality in larval flounder Electron microscopy of affected epidermal cells and transmission experiments with tissue filtrates demonstrated that the disease was caused by a herpesvirus but the agent has not been isolated in fish cell lines. On the other hand, rhabdovires infection occurrs in juvenile and production size fish with hemorrhage in the skeltal muscle and fins, congestion of the gonads, and ascites. A rhabdovirys was isolated in RTG-2 cells from the diseased flounder as a causative agent, which was designated hirame rhabdovirus (HRV) or Rahbdovirus olivaceus. HRV is serologically distinguishable from other known fish rhabdoviruses. Intensive researches on these viral diseases started in 1980th. but properties of the causative agents and infection mechanisms have not been fully investigated. This results in difficulty in controlling these diseases.

• Fisheries and Aquatic Sciences
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• v.12 no.2
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• pp.98-103
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• 2009

The potential utility of the rockbream (Oplegnathus fasciatus) $\beta$-actin 5'-upstream sequence as a regulatory element for DNA vaccination was evaluated based on in vitro and in vivo heterologous expression assays. In the in vitro transfection experiment, the efficacy of the rockbream $\beta$-actin promoter to drive the expression of a downstream lacZ gene was significantly higher (more than fourfold) than that of the human cytomegalovirus (hCMV) promoter in two fish cell lines (grunt Haemulon plumierii fin and bluegill Lepomis macrochirus fry cell lines). In contrast, the functional activity of the rockbream $\beta$-actin promoter was hardly detectable in a mammalian mouse embryonic fibroblast cell line. Rockbream skeletal muscles injected in vivo with a GFP reporter construct driven by the $\beta$-actin promoter displayed the significantly higher expression of a GFP protein (more than threefold) than did those injected with hCMV promoter driven construct. Data from this study suggest that the homologous rockbream $\beta$-actin promoter could be used as a potential regulator for DNA vaccination in this species.

• Journal of Marine Bioscience and Biotechnology
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• v.12 no.2
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• pp.108-114
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• 2020

A variety of shellfish species sold for human consumption are available for purchase in the domestic fish market. The microalgae families inhabit the ocean, where planktons supply the main nutritional resource for the growth of shellfish. Some phytoplanktons produce toxic compounds that are accumulated in shellfish and ultimately cause toxicity in humans. This article reports the cytotoxicity of commercially available shellfish species. Accordingly, hot water extract (HWE) and an aqueous fraction of 50% methanol extract (MEE-AF) showed no significant cytotoxicity on the two cell lines (i.e., HL-60 and Vero cell lines), but 50% methanol extract (MEE) in 3, 6 samples showed 50% cytotoxic effects on HL-60 cells, and 1, 4 samples showed 40%, 20% cytotoxic effects on Vero cells, respectively. In addition, their consequential dichloromethane fractions (MEE-DF) exhibited significant toxicities at the highest concentration (1,000 ㎍/ml) on HL-60 and Vero cells. Since the shellfish samples showed cytotoxicity in the dichloromethane fraction, it is possible that the dichloromethane fraction contains marine toxins. Further research will be needed to identify the toxic components from each sample.

• Journal of Genetic Medicine
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• v.5 no.1
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• pp.61-64
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• 2008

A 24-year-old female with primary amenorrhea was referred for a chromosome study. The karyotype of the patient was 46,X,der(X) under initial GTG-banding analysis. Fluorescence in situ hybridization (FISH) analysis with an LSI Kallmann (KAL) region probe [probes for Xp22.3(KAL) and CEP(X) for control] was carried out. The abnormal chromosome was KAL- and CEP(X)${\times}2$. In addition, interphase FISH analysis revealed the patient to be mosaic for two different cell lines: 90% of cells had three signals and 10% of the cells had only one signal for CEP(X). Based on these results, the karyotype of the patient was 45,X/46,X,psu idic(X)(p22.1), which is partial trisomy for Xqter${\rightarrow}$Xp22.1 and partial monosomy for Xpter${\rightarrow}$Xp22.1. This karyotype was considered a variant of Turner syndrome. In summary, Idic(X) and low-level mosaicism was successfully characterized by FISH analysis with a CEP(X) probe.

• Fisheries and Aquatic Sciences
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• v.3 no.1
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• pp.52-63
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• 2000

Non-specific reaction has been a problem in doing, especially, research and diagnosis for infectious agents. Avidin-biotin-peroxidase complex (ABC) techniques has widely been used to amplify a reaction. Photobacterium damse1a subsp. piscicdia (formerly Pasteurella piscicida) exhibited a capacity to bind with streptavidin non-specifically. The band, estimated 26 K Da in Western blotted paper, was blocked with biotin but incompletely. In an attempt to explore an involvement of the non-specific substance in attaching piscine cells, cell attachment test performed using anti- Ph. d. subsp piscicida sera raised mouse and rabbit exhibited slightly blocking effects for Mediterranean (1736) and significantly for Japanese (Sp 92144) isolate. Biotin decreased the attachment ability significantly for Sp92144 but it was not effective to 1736. Both isolates showed greatly enhanced attachment ability with poly-L-lysin. The non-specific binding substance was contained in bacterial extracellular products (ECPs). The substance was able to purified with 2-imminobiotin affinity column, the purified substance appeared to have 4 bands in silver staining, and had a carbohydrate branch. This purified substance showed cytotoxic effects selectively between 5 piscine cell lines. Moreover, it stimulated rainbow trout macrophage in terms of reduction of cytochrome cas well as yeast phagocytosis, significantly.

• Journal of fish pathology
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• v.4 no.2
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• pp.87-94
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• 1991

In the late summer of 1990 and 1991, mass mortality occured among cage-cultured grouper, Epinephelus septemfasciatus in south cost of Korea. The moribund fish didn't feed and became pale or dark chestnut colour and irregularly swimmed due to the loss of equilibrium, finally the diseased fish fell down side away on the bottom or the surface of cage showing the bent of body and died. The diseased fish showed the extensive hemorrahge in brain, the swelling of spleen and bile duct as the specific syptoms of internal organs. So the gill, skin and other organs of the diseased fish were examined for the presence of pathogenic parasites and bacteria. The parasitic Trichodina sp. were detected only from the gill lamella of the diseased fish, but these parasites seemed to be not a direct causative agents that induced the gross mortality of the cultured grouper. because these parasites were also observed in normal grouper, yellowtail, red seabream and rock bream co-cultured with the diseased grouper in same or near cages. In the viral examination, although isolation of the causative agent by the use of estabilshed cell Lines, RTG-2 and CHSE-214, was not succeed, the normal grouper inoculated intramuscularly with the filtered homogenate of the organs of the diseased fish showed the same external and internal signs with the naturally infected grouper. They died within a week. By using the naturally and the artificially infected fishes, electron microscopic observation revealed numerous hexagonal or polygonal particles in the cytoplasm of liver cells. Based on the these results, we suggest that the mass mortality of the cultured grouper would be occurred by the infection of a viral agent.

• Korean Journal of Microbiology
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• v.22 no.2
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• pp.85-90
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• 1984

Two viruses were isolated from kidney and spleen tissues of goldfish(Carasius auratus) and the ovarian fluid of chum salmon(Oncorhynchus keta). Both viruses replicated and produced cytopathic effect in EPC, CHSE-214, and CHH-1 cell lines at $15^{\circ}C$. The isolates were resistant to pH 3 and choloroform. Antiserum to infections pancreatic necrosis virus(IPNV) serotype VR 299 neutralized the infectivity of both of the isolates. Electron microscopy showed that the particles had typical IPNV particle morphology with average diameters of 55nm, This paper describes the first isolation of viruses infecting cultured fish in Korea.

• Korean Journal of Veterinary Research
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• v.46 no.1
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• pp.35-41
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• 2006

Vitellogenin (Vtg), a phospholipoglycoprotein precursor of egg yolk is synthesized and secreted from the liver in response to estrogens in female fish. Vtg is normally undetectable in the blood of male fish, but can be induced by exposure to chemicals possessing estrogenic activity. Thus, the presence of Vtg in blood of male fish can serve as a useful biomarker for assessing previous exposure to estrogenic compounds. In the present study, Vtg was abnormally expressed in Rhynchocypris oxycephalus using estradiol benzoate ($E_2$). As the result, it was found that the level of Vtg in blood from R. oxycephalus was increased by treated quantity of $E_2$ with dose-effect manner. Monoclonal antibodies were generated against Vtg of R. oxycephalus. The hybridoma were screened with an enzyme immunoassay for the production of specific anti-Vtg antibodies. Five positive cell lines with a high specificity were selected. Monoclonal antibodies against vtg of R. oxycephalus that was developed in this study, may be a useful bio-indicator for the detection of estrogenic contamination in the aquatic ecosystem.