• Journal of Microbiology and Biotechnology
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• 제20권2호
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• pp.271-280
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• 2010

Vibrio alginolyticus, a Gram-negative marine bacterium, is one of the causative agents of fish vibriosis. Its virulence factors and pathogenesis mechanism are barely known, except for some extracellular products (ECPs) that are known to be regulated by quorum sensing system. Therefore, the present study used a microarray to analyze the transcription profiles of the wild-type V. alginolyticus and a deletion mutant of luxO, the pivotal regulator in Vibrio quorum sensing systems, which resulted in the identification of a putative virulence factor, MviN. Quantitative real-time reverse transcription PCR confirmed that the transcription of mviN was upregulated in the luxO mutant when compared with wild-type, and down regulated in a luxO-con complemented strain. Furthermore, Western blotting indicated that MviN was greatly induced during the late-exponential and stationary phases of growth, indicating that the expression of MviN was cell-density dependent and quorum sensing regulated in V. alginolyticus. Meanwhile, the mviN null mutant displayed a much slower growth rate than the wild type, signifying the essential role of MviN in V. alginolyticus. Western blotting also revealed that MviN was present as an extracellular protein in V. alginolyticus. When epithelioma papulosum cyprini (EPC) cells were treated with the ECPs of the mviN mutant, no cytotoxicity was observed, whereas EPC cells treated with the wild type exhibited pathological changes, which increased with the ECPs concentration and treatment time. Therefore, the results demonstrated that MviN is a LuxO-regulated ECPs component and involved in the pathogenicity of V. alginolyticus.

• 한국어병학회지
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• 제18권1호
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• pp.67-80
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• 2005

미꾸라지 (mudloach, Misgunus mizolepis)의 간으로부터 클로닝한 phosphoinositide-specific phospholipase C$\delta$ (ML-PLC$\delta$)를 대장균 (E. coli)에서 과발현시켜 만든 재조합 ML-PLC$\delta$와 미꾸라지 간 조직으로부터 직접 정제한 ML-PLC$\delta$의 생화학적 특성을 비교분석하였다. 우선, pET28a vector (Novagen)를 이용하여 E. coli BL21(DE3)에서 과발현된 재조합 ML-PLC$\delta$은 $Ni^{2+}$-NTA affinity 크로마토그래피 및 gel filtration 칼럼에 의해서 정제되었다. 미꾸라지 간 조직으로 ML-PLC$\delta$는 open heparin 칼럼 및 분석용 heparin 칼럼등을 통하여 부분 정제하였다. 두개의 재조합 및 wild ML-PLC$\delta$는 phosphatidylinositol 4,5-bis-phosphate ($PIP_2$)에 대한 농도 의존적 PLC 활성을 보여주었고, 그 활성은 포유류 PLC$\delta$ 효소와 유사하게 칼슘 농도에 의존적인 활성을 나타내었다. 재조합 및 wild ML-PLC$\delta$는 각각 pH 7.0 및 7.5에서 가장 큰 PI-가수분해 활성을 나타낸다는 사실을 알 수 있었다. 게다가, 재조합 및 wild ML-PLC$\delta$는 sodium doecylcholate (SDC) 및 phosphatidylethanolamine (PE), phosphatidylcholine (PC)와 같은 지질류에 대하여 농도의존적인 활성을 나타내나, spermine과 같은 polyamine류의 존재하에서는 농도 의존적으로 PLC 활성이 감소됨을 알 수 있었다. 미꾸라지 각 기관들의 ML-PLC$\delta$의 발현양상 및 양등을 측정하여 보았을 때 ML-PLC$\delta$는 포유류 PLC$\delta$와 마찬가지로 다양한 형태의 PLC$\delta$가 존재함을 알 수 있었다. 이와 같은 결과들로 미루어서 미꾸라지로부터 얻은 ML-PLC$\delta$는 포유류의 PLC$\delta$ isozymes과 유사한 형태의 생화학적 특성을 가지나, 포유류 PLC$\delta$1과 PLC$\delta$3 isozyme의 생화학적 특성을 함께 가짐을 알 수 있었다.

• 한국어병학회지
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• 제26권1호
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• pp.39-44
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• 2013

자연산 다묵장어, Lethenterone reissneri에서 발생한 물곰팡이병의 원인체를 형태학적 및 유전학적 연구를 통하여 동정하였다. 분리균주는 격벽이 없는 균사를 형성하였고, 가늘고 긴 모양의 유주자낭 내에 활발한 운동성을 보이는 유주자가 여러 줄로 배열되었다가 유주자낭의 선단에서 휴면하지 않고 한꺼번에 방출되는 특징을 보였다. 한편 유성생식기관인 조란기와 조정기는 관찰되지 않았다. 또한 분리균주의 ITS sequence를 분석하고 Saprolegnia 속의 다른 분리균주와 유전학적 상관관계를 조사한 결과, S. parasitica가 속해있는 clade I에 분류되는 것을 확인할 수 있었다. 무성생식기관의 형태, 유주자 방출방식 및 ITS sequence의 phylogenetic analysis에 근거하여 자연산 다묵장어에서 관찰된 물곰팡이병의 원인체는 S. parasitica인 것으로 동정되었다.

• 한국어병학회지
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• 제22권1호
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• pp.15-21
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• 2009

White spot syndrome virus (WSSV) is a strong causative agent for high mortality in cultured and wild shrimps. From this study, the WSSV prevalence in marine organisms around shrimp farm as well as live feed-mediated transmission of WSSV to farmed shrimps were investigated. Based on nested-PCR method, WSSV was detected in wide array of marine organisms including Perinereis aibuhitensis (81.3% of prevalence rate, 13/16), Enedrias fangi (100%, 16/16), Ruditapes philippinarum (20%, 2/10), crab larvae (100%, 10/10), copepoda (30%, 3/10), Periophthalmus modestus (50%, 5/10), Pachygrapsus crassipes (10%, 1/10), Helice tridens (20%, 2/10) and Neomysis sp. (70%, 7/10). On the other hand, WSSV was not detected in Bullacta exarata, Uca arcuata, and Reishia clavigera. The percent prevalence of WSSV in wild shrimps, Fenneropenaeus chinensis was only 6%, but markedly increased up to 56% after a feeding trial using polychaete worms for one month, indicating that the live feed is one of significant carriers of WSSV to shrimps under practical farming conditions.

• Fisheries and Aquatic Sciences
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• 제22권6호
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• pp.12.1-12.8
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• 2019

Background: Wild fish populations stock is continuously diminishing in the Indo-Ganges river basin, and the population status of most fishes is unidentified. The identification of the population status and the conservation of commercially important and endemic wild fish populations in this region are crucial for the management. The aim of this paper was to identify the population status of Cirrhinus reba, a promising aquaculture but vulnerable species in the Indo-Ganges river basin in Bangladesh. Methods: C. reba samples were collected from four isolated populations of the Brahmaputra (n = 30), the Padma (33), the Karatoya (31), and the Jamuna Rivers (30) in Bangladesh, and the population status was evaluated using morphometric and landmark comparisons. Data were analyzed with the Kruskal-Wallis test, univariate analysis, discriminant function analysis, and the formation of a dendrogram. Results: Three meristic characters (Pectoral fin rays, caudal fin rays, scale in lateral lines), four morphometric characters (head length, pre-orbital length, post-orbital length, maximum body depth), and truss measurement (4-7) were significantly different among the stocks. The step-wise discriminant function analysis retained 15 variables from morphometric and landmark measurements that significantly differentiated the populations based on the constructed DFI and DFII. Discriminate function analysis also showed that 91.2% of the original groups were classified into their correct samples. The cluster analysis of Euclidean distances placed the Jamuna population in one cluster and the Brahmaputra, the Padma, and the Karatoya populations in the second one. Conclusion : Morphological differences among the stock were probably due to different ancestral origin. This is the first report about population status of C. reba in their natural habitat of the Indian subcontinent. Further genetic studies and the evaluation of environmental impact on C. reba populations in Bangladesh are suggested to support our findings.

• 한국어류학회지
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• 제32권3호
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• pp.130-135
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• 2020

양식업에서 인기 있는 어종 중 하나인 동자개를 자연산과 양식산을 구분하여 생식소 발달을 관찰하였다. 자연산과 양식산 동자개는 2018년 4월에서 10월에 채집하여 수컷의 생식소 지수, 비만도지수 및 생식소 발달을 연구하였다. 광학현미경으로 관찰된 월별 생식소 발달과 GSI를 기반으로 생식소 발달 단계는 자연산과 실외 사육장은 4월 성숙기, 5월 산란기, 6월에서 7월 퇴화기, 8월에서 10월 휴지기로 관찰되었다. 실내 사육장은 4월에서 6월까지 성숙기, 7월에서 8월 산란기, 9월 퇴화기, 10월 휴지기로 구분되었다. GSI 값은 자연산과 실외 사육장은 5월에 가장 높은 값을 보였고, 실내 사육장은 9월에 가장 높은 값을 나타내 차이를 보였다. CF는 자연산과 양식산 사이에 유의한 차이가 없었으나 자연산은 6월에 가장 높은 값을 보였으며, 실외 사육장은 11월에 가장 높은 값을 보였다. 본 실험 결과, 실외 사육장 동자개 수컷을 인공 수정에 사용하는 것이 적합하며, 자연산 생식주기와 같은 5월에 인공 수정을 하는 것이 효율적일 것으로 판단된다.

• 한국어류학회지
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• 제21권sup1호
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• pp.29-37
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• 2009

The tiny fragment of freshwater body is providing home for huge biodiversity and resources for the existence of human. The competing demand for freshwater have been increased rapidly and it caused the declination of biodiversity in recent decades. Unlike the natural process of extinction in gradual progress, the current species extinction is accelerated by human activity. As a result many fish species are already extinct or alive only in captivity in the world and about fifty eight animal species are in endangered in Korea including eighteen freshwater species. Conservation of biodiversity is the process by which the prevention of loss or damage is attained, and is often associated with management of the natural environment. The practical action is classified into in-situ, or ex-situ depending on the location of the conservation effort. Recovery means the process by which the status of endangerment is improved to persist in the wild by re-introduction of species from ex-situ conservation population into nature or translocation of some population. However there are a lot of restrictions to complete it and successful results are known very rare in case. In this article the authors explore some strategies for conservation and restoration of freshwater fish species conducted in Korea for few years. The major causes are discussed in relation with the decline of freshwater fish diversity during few decades and some strategies are evaluated to advance the process of conservation. A study on the Korean bullhead, Pseudobagrus brevicorpus, is introduced as a case for ex-situ conservation and restoration in freshwater ecosystem.

• Fisheries and Aquatic Sciences
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• 제14권4호
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• pp.403-410
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• 2011

Scartelaos gigas is an amphibious mudskipper species that inhabits mud flats in Korea, China, and Taiwan. This fish is at risk of extinction because of its very restricted habitat and overexploitation. Information about this fish's reproductive characteristics is needed for species conservation. The sexual maturity and early life history of S. gigas were investigated through histological methods and direct observation of eggs in the wild, respectively. In total, 560 individuals of S. gigas were collected with the aid of fishermen from March 2003 to October 2003 at Jung-do Island, southwest Korea. Through microscopic observations of gonadal development, it was determined that S. gigas of both sexes were immature in April, but began to reach maturity in May, and were then fully mature by June, which was maintained until July. In August, some female fish developed early oocytes, but by September oocytes were observed to have degenerated and had been absorbed. Spawned eggs were elliptical and had an average size of 1.37 mm (long axis) by 0.69 mm (short axis). The newly hatched larvae (3.03 mm total length, TL) had an open mouth and anus, two melanophores near the anus, and one large melanophore between the 18th and 19th myomeres. The larvae (3.18 mm TL) showed absorption of the yolk and oil globule within 5 days after hatching and became prelarvae. This species should be considered vulnerable or conservation-dependent, and thus parental fish need to be protected from fishermen during the main spawning season (June).

• 한국어병학회지
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• 제17권1호
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• pp.1-10
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• 2004

해수 환경 중의 VHSV 분포 조사를 위하여, 2003년 2월에서 5월까지 동해 및 남해안 인근 해역에 서식하는 자연산 해수 어류를 채집하였다. 바이러스 분리용 시료는 9종의 해수 어류로서 42개의 시료를 분석하였다. 각 시료의 조직여과액을 Epithelioma papulosum cyprini (EPC) cells에 접종한 결과, 고등어 Scomber japonicus와 숭어 Mugil cephalus에서 바이러스가 분리되었으며 이들 바이러스는 RT-PCR법을 이용하여 VHSV로 동정할 수 있었다. 고등어에서 VHSV가 검출되었다는 기보고는 있지만, 숭어에 대해서는 본 연구에서 처음으로 VHSV를 분리하였다. 자연산 어류에서 유래된 세 개의 VHSV isolates에 대한 glycoprotein gene을 분석한 결과, 이들 바이러스는 양식 넙치에서 질병을 유발하는 VHSV isolates와 유사하며 모두 genogroup I (American type)에 속하는 것으로 밝혀졌다.

• Fisheries and Aquatic Sciences
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• 제21권11호
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• pp.34.1-34.13
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• 2018

Sexual maturity ($L_{50}$), the length at which 50% of fish in a size class are mature, is a key aspect of domestication of new fish species because it guides the procedure for identification of appropriate broodstock size for artificial spawning. In this study, the $L_{50}$ was determined for 1083 Barbus altianalis samples obtained from Lake Edward and the Upper Victoria Nile. Gonads of freshly killed samples were examined macroscopically and verified with standard histological procedures for the maturation stages that were used to determine $L_{50}$. Oocytes and spermatogenic cell sizes were compared for fish obtained from both water bodies. Results indicated that there were no variations in macro gonad features observed for fish from Lake Edward and Upper Victoria Nile. Similarly, there were no significant differences in oocyte sizes (P > 0.05) between the two populations but significant differences in spermatogenic cell sizes were noted (P < 0.05) except for spermatozoa (P > 0.05). This however did not suggest peculiar differences between the two populations for staging the gonads. Consequently, no staging variations were suggested for both populations in determination of $L_{50}$. Sexual maturity was found in the same class size of fork length (FL) 20-24.9 cm and 35-39.9 cm for males and females from both water bodies, respectively. At this FL, however, males were too small, and for good selection of vigor broodstocks for spawning and conservation purposes, they are better picked from class size of 30-34.9 cm FL and above. These findings were crucial for integration of appropriate breeding size in spawning protocol by farmers and fisheries scientists conserving wild B. altianalis populations.