• Fisheries and Aquatic Sciences
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• 제8권3호
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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.

• 한국어병학회지
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• 제5권1호
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• pp.1-7
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• 1992

CHSE-214 무지개송어 세포주는 10% fetal bovine serum과 2mM-glutamine이 첨가된 Eagle's 기본배지에서 성장하였다. CHSE-214 세포주의 최적온도는 $20^{\circ}C$였다. IPN바이러스는 CHSE-214 세포주에서 복제되었으며, 세포변성효과를 나타내었다. 또한 IPN 바이러스의 감염시간에 따른 감염증상을 역위상차 현미경으로 관찰하였다. 감염 6-12시간 후 세포는 정상세포와 비슷하였다. 감염 18-24시간 후 세포는 일부가 정상 세포보다 더 둥근형태로 되었고, 일부세포는 세포배양용 플라스크 바닥에서 떨어져 부유상태로 되었다. 감염 30시간 후 세포는 더더욱 비정상적인 형태로 되었다. 감염 48~68시간 후 감염된 세포는 파열되었고, 아주 높은 세포 병변 효과를 나타내었다.

• 한국어병학회지
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• 제6권2호
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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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• 제12권2호
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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.

• 한국해양바이오학회지
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• 제12권2호
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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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• 제5권1호
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• pp.61-64
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• 2008

24세의 무월경을 주소로 내원한 여성에게서 세포유전학적 검사를 시행하였다. GTG-분염법 결과, 환자의 핵형이 46,X,der(X)로 관찰되어 LSI Kallmann(KAL) 형광소식자[probes for Xp22.3 (KAL)과 CEP(X) for control]로 FISH 분석을 시행하였다. 비정상 X 염색체에서는 KAL (Xp22.3)에 대한 형광이 보이지 않았고, CEP(X)에 대한 형광이 두 개씩 관찰되었다. 간기세포 FISH 분석 결과, CEP(X) 형광소식자에 대해 분석한 세포의 90%에서 세 개의 형광이, 10%의 세포에서 하나의 형광이 관찰되어 두가지 cell line이 혼재되어 있는 모자이시즘을 확인하였다. 이들 결과들을 통해 환자의 핵형이 45,X/46,X,psu idic(X)(p22.1)이며, 이는 Xqter${\rightarrow}$Xp22.1 부분은 중복되어 있고, Xpter${\rightarrow}$Xp22.1 부분은 결실된 상태로 터너 증후군의 변형된 형태임을 판명하였다. Idic(X)와 낮은 비율의 모자이시즘을 확인하는데 CEP(X) 형광소식자로 FISH를 시행하는 것이 유용하였다.

• Fisheries and Aquatic Sciences
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• 제3권1호
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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.

• 한국어병학회지
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• 제4권2호
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• pp.87-94
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• 1991

1. 자연감염(自然感染) 능성어는 외관적으로 먹이를 먹지 않고 분색이 옅은 밤색이나 짙은 밤색으로 변하며 몸이 휘어져 힘없이 가두리 바닥이나 수면에 누워서 폐사(斃死)하였는데, 해부(解剖)를 해보면 뇌출혈(腦出血)과 비장(脾臟) 및 담관(膽管)이 팽대(膨大)되어 있었다. 2. 자연감염어(自然感染魚) 아가미에서 Trichodina sp. 기생충(寄生蟲)이 다수 검출(檢出)되지만 대량(大量) 폐사(斃死)의 직접(直接) 원인생물(原因生物)은 아니며, 또한 병어(病魚)의 장기(臟器)에서 우점적(優占的)으로 분리(分離)되는 Vibrio속(屬) 세균(細菌)은 병원성(病原性)이 없는 해수상존(海水常存) Vibrio 균(菌)이었다. 3. RTG-2 및 CHSE-214 어류(魚類) 주화세포(株化細胞)에서 바이러스는 분리(分離)되지 않았지만, 병어(病魚)의 장기마쇄여과액(臟器磨碎濾過液)으로 인위감염(人爲感染)시켰을 때 자연감염어(自然感染魚)와 유사(類似)한 증상(症狀)을 나타내면서 폐사(斃死)하였다. 4. 자연감염어(自然感染魚) 및 인위감염어(人爲感染魚)의 간(肝) 세포질내(細胞質內) 공포부위(空胞部位)에 크기가 45-60nm 정도되며 육각형(六角形) 내지 다각형모양(多角形模樣)을 한 virion들이 무수히 존재하였다. 5. 따라서 1990년(年)과 1991년(年) 고수온기(高水溫期)에 우리나라 남해안(南海岸) 일원 해상가두리 양식장에서 사육중이던 능성어가 대량(大量) 폐사(斃死)한 것은 바이러스성(性) 질병(疾病)에 의한 것으로 생각된다.

• 미생물학회지
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• 제22권2호
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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.

• 대한수의학회지
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• 제46권1호
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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.