• 한국어병학회지
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• 제12권2호
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• pp.107-113
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• 1999

능성어의 바이러스성 신경괴사증 바이러스(fish nodavirus)의 병원성을 능성어 및 다른 해산어류를 대상으로 인위감염법에 의해 조사한 결과, 능성어 바이러스성 신경괴사증 바이러스는 능성어에서는 소형어(27 g)부터 대형어(104 g)까지 제사를 일으켜 강한 병원성을 나타냈지만, 실험에 사용한 크기의 방어(537 g), 참돔(207 g), 돌돔(43 g), 넙치(41 g), 자주복(27 g) 및 조피볼락(94 g)에는 폐사를 일으키지 않아 병원성을 나타내지 않았다. 그리고 사육수온별 능성어에 대한 바이러스의 병원성은 수온 $15^{\circ}C$에서는 병원성을 나타내지 않았고, 수온 $20\sim28^{\circ}C$에서는 수온이 높을 수록 병원성을 강하게 나타냈다. 그리고 바이러스는 근육주사, 복강 주사 및 경구감염뿐만 아니라 침지감염과 혼합사육감염에 의해서도 쉽게 능성어에 감염이 되지만, 감염 후 생존한 능성어에 대해서는 바이러스를 재감염시켜도 폐사가 일어나지 않았다.

• Journal of Microbiology and Biotechnology
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• 제25권10호
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• pp.1761-1767
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• 2015

Sevenband grouper, Epinephelus septemfasciatus, is becoming an important aquaculture species in Korea. However, viral nervous necrosis disease is a large problem causing mass mortality in sevenband grouper aquaculture. Recombinant protein vaccines are one of the best methods to reduce these economic losses. However, the cell-based expression method mainly produces inclusion bodies and requires additional procedures. In this study, we expressed a recombinant viral coat protein of sevenband grouper nervous necrosis virus (NNV) using a cell-free protein synthesis system. The purified recombinant NNV coat protein (rNNV-CP) was injected into sevenband grouper at different doses followed by a NNV challenge. Nonimmunized fish in the first trial (20 μg/fish) began to die 5 days post-challenge and reached 70% cumulative mortality. In contrast, immunized fish also starting dying 5 days postchallenge but lower cumulative mortality (10%) was observed. Cumulative morality in the second trial with different doses (20, 4, and 0.8 μg/fish) was 10%, 40%, and 50%, respectively. These results suggest that rNNV-CP can effectively immunize sevenband grouper depending on the dose administered. This study provides a new approach to develop a recombinant vaccine against NNV infection for sevenband grouper.

• 한국어류학회지
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• 제31권4호
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• pp.235-240
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• 2019

자바리와 대왕자바리 2종을 대상으로 MS-222 농도 및 수온에 따른 마취효과 실험을 수행하였다. 농도는 100, 150, 200 그리고 250 ppm을, 수온은 18, 22, 26 그리고 30℃에서 마취 및 회복시간을 조사하였다. MS-222 농도와 수온이 높아질수록 마취시간이 짧아지는 경향을 보였으며, 회복시간은 농도와는 비례하지만 수온과는 반비례하는 경향을 보였다. 회복시간은 22℃ 이하에서는 큰 차이가 없었다. 자바리의 최적 마취조건은 회복시간이 가장 짧은 30℃와 100 ppm, 대왕자바리의 경우 30℃ 150 ppm이 회복시간이 100 ppm과 유의적으로 차이가 없으면서 마취시간은 유의하게 짧아 가장 적합했다. 이원배치분산분석 결과 자바리와 대왕자바리의 종 간 유의한 차이가 있었으나, 종과 농도 그리고 종, 농도, 수온에 대한 상호작용 효과는 없었다.

• 한국어류학회지
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• 제31권1호
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• pp.23-29
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• 2019

이번 연구에서 바리과 어종의 새로운 교잡종을 생산하기 위한 연구의 일환으로 능성어 (Hyporthodus septemfasciatus) ♀${\times}$대왕바리 (Epinephelus lanceolatus) ♂ (CGGG)의 발생 양상, 부화율, 기형 발생률 그리고 부화 소요시간을 조사하고, 교잡종의 모계어종의 순종 (능성어 ♀${\times}$♂, CG)과 비교하였다. CGGG와 CG 수정란은 자외선 살균 해수가 채워진 5-L 용량의 비이커 9개에 3,000~5,000개/비이커의 밀도로 수용되었으며, 발생 기간 중 수온은 $23.5{\sim}24.8^{\circ}C$ 그리고 염분은 32.1~32.8 ppt 였다. CGGG의 수정률과 부화율은 각각 $69.4{\pm}1.5%$와 $59.0{\pm}5.1%$로서 CG에 비해 유의하게 낮았다 (p<0.05). CGGG는 비균등 난할, 배체 체형 기형 그리고 부화자어의 척추 기형이 높게 출현한 것을 제외하고, CG와 동일한 난발생 양상을 보였다. CGGG의 부화까지 소요 시간은 CG에 비해 약 2시간 느렸다(p<0.05). 이상의 결과에서 CGGG는 수정률, 부화율 그리고 기형률 등 발생능력이 CG에 비해 낮았지만 부화자어의 대량생산이 가능할 것으로 판단된다.

• 한국해양생명과학회지
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• 제2권1호
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• pp.1-6
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• 2017

본 연구의 목적은 능성어와 자바리의 정자동결을 위한 간편한 실험법개발이다. 희석제와 동해방지제가 정자동결에 미치는 효과를 파악하고자 운동성성과 생존율을 조사하였다. 동결실험에서 300 mM glucose와 15% dimethylsulfoxide (DMSO)를 희석제와 동해방지제로 각각 사용하였다. 동결실험결과, 능성어 정자는 동결 5개월 후 60% 이상의 운동성을 보였고, 자바리 정자는 동결 5개월후 90% 이상의 운동성을 보였다.

• 수산해양기술연구
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• 제35권4호
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• pp.391-396
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• 1999

It is well known that fish hear a wide frequency range of sound using both laterial line and inner ear, and the sound is probably the most effective channel for long-range communication underwater than any other stimulus.The auditory threshold of svenband grouper, Epinephelus septem fasciatus was determined by conditioning method using a sound coupled with electric shock in a tank. The audio-signals and electric shock used in the experiment were pure tones of 100 to 1000Hz and DC 6n.The response of fish to the sound was observed by ECG intervals from precordial leads method (V1·V2 method). The auditory threshold curve for sevenband grouper was V-shaped, with most sensitive threshold at 350Hz in the range of 100~1000Hz. The Sevenband grouper were more sensitive to sounds of 200~400Hz than to adjacent frequencies.

• 한국어류학회지
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• 제26권1호
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• pp.70-73
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• 2014

Interspecific hybrids between tiger grouper Epinephelus fuscoguttatus and giant grouper E. lanceolatus were genetically identified based on the partial sequence analysis of mitochondrial cytochrome c oxidase I (COX I) gene and nuclear recombination activating gene 2 (RAG 2) gene. Out of 585 base positions of RAG 2, a total of five nucleotide substitutions were detected between the two parental species (E. fuscoguttatus and E. lanceolatus). The hybrids had two distinct types of RAG 2 sequences corresponding to those of both parental species. Mitochondrial COX I gene sequencing showed that hybrids had sequences identical to E. fuscoguttatus. Molecular data clearly demonstrate that hybridization does occur between E. fuscoguttatus and E. lanceolatus, but with E. fuscoguttatus as the maternal parent.

• 한국어병학회지
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• 제34권1호
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• pp.111-115
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• 2021

Immunoglobulin M (IgM) of sevenband grouper (Epinephelus septemfasciatus) was purified by mannan-binding protein (MBP) affinity column. The purified IgM had an apparent molecular weights of 76 (heavy chain) and 28 (light chain) kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Eight hybridoma clones secreting monoclonal antibodies (mAbs) against sevenband grouper IgM were established. Antibody detection enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the 8 mAbs revealed that optical density (OD) values were clearly different between sera from BSA-immunization and non-immunization of sevenband grouper. These results suggest that the produced mAbs in this study are specifically reacted with IgM of sevenband grouper.

• 한국발생생물학회지:발생과생식
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• 제18권3호
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• pp.127-131
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• 2014

We investigated the androgenic effects of $17{\alpha}$-methyltestosterone (MT) on gonadal sex reversal in juvenile red spotted grouper Epinephelus akaara. The fish were immersed in $17{\alpha}$-MT at 1 and 5 mg/L. Treatment method of $17{\alpha}$-MT was once weekly for 4 and 8 weeks. Fish were sampled at 12 months after end of the treatment period in order to histological analysis. At the initiation of an experiment (70 day after hatching), juvenile red spotted grouper have the paired primordial gonads with somatic cells bellow kidney in the posterior portion of the body cavity. Formation of ovarian cavity indicates that the ovarian differentiation beginning at 70 DAH in red spotted grouper. At 12 months after end of the treatment period, control group, $17{\alpha}$-MT 1 mg/L treatment group for 4 and 8 weeks, and $17{\alpha}$-MT 5 mg/L treatment group for 4 weeks were all female. However, sex-changed males without ovarian cavity were observed in the $17{\alpha}$-MT 5 mg/L treatment group for 8 weeks. In grouper, we firstly reported that the red spotted grouper be able to induce the primary males by hormone treatment prior to gonadal sex differentiation.

• Journal of Microbiology and Biotechnology
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• 제31권8호
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• pp.1088-1097
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• 2021

Grouper nervous necrosis virus (GNNV) infection causes mass grouper mortality, leading to substantial economic loss in Taiwan. Traditional methods of controlling GNNV infections involve the challenge of controlling disinfectant doses; low doses are ineffective, whereas high doses may cause environmental damage. Identifying potential methods to safely control GNNV infection to prevent viral outbreaks is essential. We engineered a virus-binding bacterium expressing a myxovirus resistance (Mx) protein on its surface for GNNV removal from phosphate-buffered saline (PBS), thus increasing the survival of grouper fin (GF-1) cells. We fused the grouper Mx protein (which recognizes and binds to the coat protein of GNNV) to the C-terminus of outer membrane lipoprotein A (lpp-Mx) and to the N-terminus of a bacterial autotransporter adhesin (Mx-AIDA); these constructs were expressed on the surfaces of Escherichia coli BL21 (BL21/lpp-Mx and BL21/Mx-AIDA). We examined bacterial surface expression capacity and GNNV binding activity through enzyme-linked immunosorbent assay; we also evaluated the GNNV removal efficacy of the bacteria and viral cytotoxicity after bacterial adsorption treatment. Although both constructs were successfully expressed, only BL21/lpp-Mx exhibited GNNV binding activity; BL21/lpp-Mx cells removed GNNV and protected GF-1 cells from GNNV infection more efficiently. Moreover, salinity affected the GNNV removal efficacy of BL21/lpp-Mx. Thus, our GNNV-binding bacterium is an efficient microparticle for removing GNNV from 10‰ brackish water and for preventing GNNV infection in groupers.