• Fisheries and Aquatic Sciences
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• 제7권1호
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• pp.10-15
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• 2004

A total of 52 pathogenic Vibrio strains was isolated from the Gwangan Beach during summer in 2003. The isolated vibrios were composed of 6 different species: V. parahaemolyticus, V. cholerae non O1, V. fluvialis, V. vulnificus, V. alginolyticus, and V. mimicus. V. parahaemolyticus was most predominant as $46\%$ (24/52), V. cholerae non O1 was the second with $23\%$ (12/52), and V. fluvialis was the third with $17\%$ (9/52). Among the isolated strains, 22 strains showed hemolytic, proteolytic or ureolytic activity. Eight strains showed both hemolysin and protease activities, and either 6 strains showed only hemolysin activities and 7 strains only protease activities. Only one strain of V. parahaemolyticus isolates showed urease activity. The urease-positive V. parahaemolyticus strain (V. parahaemolyticus S25) showed the same biochemical characteristics as the reference strain, V. parahaemolyticus KCTC 2471 (urease­negative) except for urease production. To compare the degree of virulence of Vibrio strains having different pathogenic factors, hemolysin, protease, or urease-positive strains were injected into groups of 10 each of ICR mice (7- to l0-week-old male). The lethal rate of urease-positive V. parahaemolyticus S25 was significantly high, being $70\%$. Protease-positive strains showed $40-60\%$ of lethal rate. Hemolysin-positive strains showed no mortality, similar to non-pathogenic V. parahaemolyticus KCTC 2471 and V. parahaemolyticus FM12.

• 한국수산과학회지
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• 제34권6호
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• pp.611-616
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• 2001

통영항 내의 해수 및 저질의 물리 화학적 해양환경 특성과 해수의 위생지표세균을 비롯한 병원성 세균의 시험결과가 횟집의 수족관의 세균학적 수질에 적합한지에 대하여 검토하였다. 조사기간중 통영 항 해수는 수온 $6.8\sim25.2^{\circ}C$, 투명도 $1.0\sim2.5\;m$, 화학적산소요구량 $1.79\~2.41\;mg/L$, 용존산소 $5.7\~10.1\;mg/L$, 용존질소 $6.59\sim10.53{\mu}g-at/L$, 인산염 $0.56\sim1.01{\mu}g-at/L$, chlorophyll-a는 $1.21\sim9.54 mg/m^3$ 범위였으며 염분농도는 $30.11\sim31.41\%$였다. 통영항 해수의 생균수는 조사기간 중 mL당 log수가 $4.2\~6.8$($3.0\times10^4CFU/mL \sim6.9\times10^6CFU/mL$) 범위에 평균 5.3이었다. 월별로는 $1\~3$월이 적었고, $7\~9$월이 많았다. 통영항 내 조사해수의 $84.3\%$와 $93.1\%$가 패류수출해역 수질기준 중 total coliform group과 fecal coliform group의 기준치를 각각 초과하였다. 대장균군형은 Escherichia coli group이 $66.1\%$를 차지하였고, 그중 Type I 이 $56.5\%$, Citrobacter freundii group이 $11.0\%$, Enterobacter aerogenes group이 $9.69\%$, 기타 29균주로 $13.3\%$에 해당하였다. 그런데 전형적인 온혈동물의 장관유래균인 E. coli type I 이 전체의 $56.5\%$를 차지하였다. 통영항내 해수에서 Salmonella sp.나 Shigella sp. 및 Vibrio cholerae O1은 검출되지 않았으나 V. parahaemolyticus, V. vulnificus 및 V. cholerae non-O1 등의 검출율은 $10.4\sim35.4\%$에 달하였다.

• Fisheries and Aquatic Sciences
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• 제7권2호
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• pp.58-63
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• 2004

Five urease-positive Vibrio parahaemolyticus strains were isolated from Kamak Bay in Yeosu in 2002 and 2003. V. parahaemolyticus YKB4 and YKB14 were isolated from seawater, YFB20 from black rockfish (Sebastes schlegeli), and YFO2l and YFO22 from olive flounder (Paralichthys olivaceus). The five urease-positive strains (YKB4, YKB14, YFB20, YFO21, and YFO22) did not show hemolysin and protease activity, while they did alter in color (to red) as the bacteria grew in the urea broth medium. All samples showed identical biochemical characteristics as a reference strain, V. parahaemolyticus KCTC2471, except in urease production. The five urease-positive strains showed urease activities at a mid stationary phase, and their activity was maximal in the late stationary phase of their culture supernatant. The addition of urea to the Luria-Bertani (LB) broth medium significantly affected the initial production of urease of V. parahaemolyticus isolates. Mortality by urease-positive V. parahaemolyticus YKB4, YKB14, YFO2l, and YFO22 was significantly high, being$60-80\%$, while YFB20 only reflected a rate of $20\%$. Protease-positive V. parahaemolyticus FM39 and FM50 showed a $40\%$ and $60\%$ mortality rate, respectively. However, hemolysin-positive V. parahaemolyticus had no mortality, like the non-pathogenic V. parahaemolyticus KCTC2471, while V. vulnificus resulted in a $40\%$ mortality rate. Injection with urease-positive V. parahaemolyticus strains showed mortality within 12 hrs in mice, and the strains could be isolated from the dead mice.

• Preventive Nutrition and Food Science
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• 제4권1호
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• pp.18-22
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• 1999

Flavor characteristics of raw Cordyceps militaris significatntly different from those of dried one. In the case of raw Cordyceps militaris , major flavor components were composed of 5 alcohols, 3 ketones, 4 phenols, 9 alkanes , and 3 alkenes. The major alcohol was 1-octen-3-ol(22.56%, 1147.3% ng/ml), which contributed to the characteristic green flavor. Ketones (3-ocatone, inparticular )were present in the highest concentration in raw Cordyceps militaris . In contrast, major flavor components of dried Cordyceps militaris were composed of 4 alcohols, 4 ketones, 3 furans, 4 pyrizines, 2 dithiazines, 5 phenols , 8alkenes , 17 alkanes, and 8 fatty acids. Dried Cordyceps militaris had unique sweet aroma of sesame as wella s a milky flavor. Green or fruit flavor were rarely detected . In alkanes , 10 cosanes, component fo wax were present. Typical flavor components of alkanes such as $\beta$-caryophyllen and Δ-cadinene were also detected. Fatty acids of dried Cordyceps militaris ranged from myristic acid (14 :0) to linoleic acid (18 ; 2). The sweet aroma of dried Cordyceps militaris was mostly due to pryazines, dithaiazines, and furans. Two dithaizines were identified and characteristics of these flavor components was a roasted bacon flavor. Strong antibacterial acitivity was observed toward Vibrio spp. such as V. vulnificus, V.cholerae, V. parahaemlyticus. Relatively high antibacterial acitivity was shown toward Bacillus subtilis , B,cereus, Staphyllococcus aureus, and Corynebacterium xerosis.

• 한국수산과학회지
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• 제29권5호
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• pp.557-566
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• 1996

병원성 비브리오균은 생선회를 즐겨 먹는 우리나라의 식습관 때문에 여름철 식중독 원인 세균 중에서 제일 빈도가 높은 세균이다. 특히 매년 여름철이면 콜레라 파동으로 활선어 판매금지 조치 등으로 생산어민 및 생선 횟집 경영자들의 경제적 손실은 매우 크다. 더구나 콜레라균이 해수에서 동물성 플랑크톤에 부착하여 월동한다는 보고도 있어서 실제로 우리 나라 연안 해수에서 콜레라균이 플랑크톤에 부착하여 월동할 가능성이 있는지를 검토한 연구 결과를 보고하는 바이다. 1. 우리 나라 남해안의 해수, 어패류 및 동물성 플랑크톤에서 V. parahaemolyticus, V. vulnificus, V. cholerae non O1, V. mimicus는 검출되었으나 V. cholerae O1은 검출되지 않았다. 2. 동물성 플랑크톤은 해수에서 각종 병원성 비브리오균의 증식이나 부착, 생잔율에 긍정적 영향을 미치고 있었다. 3. 병원성 비브리오균의 동물성 플랑크톤에의 부착율은 대장균에 비하여 높은데 염분 농도 $5\%_{\circ}$ 일 경우에는 $60\%$이상으로 높았으나, 염분 농도가 증가할수록 비례적으로 흡착율은 감소하였으며 염분 농도 $20\%_{\circ}$이상, pH 8.0 이상에서는 급격히 감소하였다. 4. 병원성 비브리오균은 $25^{\circ}C$의 해수에서는 플랑크톤이 존재할 때가 그렇지 않을 때 보다 균의 생잔율이 크게 나타났으나, $0^{\circ}C$에서는 플랑크톤의 존재 유무에 관계없이 $2\~3$일 이내에 균수가 급격히 감소되었다. 5. 미세 조류나 사멸시킨 동물성 플랑크톤의 존재가 병원성 비브리오균의 증식이나 생존에는 영향을 미치지 못하였다.

• 한국미생물·생명공학회지
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• 제44권3호
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• pp.355-362
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• 2016

수해양성 병원성 미생물로 알려진 Vibrio anguillarum으로부터 mannose-1-phosphate를 mannose-6-phosphate, glucose-1-phosphate를 glucose-6-phosphate로 가역적으로 변환시키는 phosphomannomutase/phosphoglucomutase (pmm/pgm)의 유전자를 sequencing하여 1338 bp의 open reading frame (ORF)을 밝혔다. 이는 446개의 아미노산을 포함하며 47,625 Da을 가지고 있다. 보고된 다른 Vibrio sp.의 pmm/pgm 유전자와 상동성을 비교하였을 때 V. mimicus V. vulnificus, V. splendidus, V. harveyi와 92.3%, 91.4%, 89.9%, 89.9%에 해당하는 상동성을 지니고 있었다. 증폭된 목적 유전자를 pET-28a(+) vector에 연결하여 대장균에서 단백질의 대량발현을 유도하였으며 이는 주로 soluble한 상태로 나왔다. Soluble fraction을 Ni-NTA column chromatography로 정제하여 약 50 kDa의 단백질을 얻었고 이는 주로 mannose-1-phosphate를 이용하는 효소로 확인되었으며 Mg²⁺ 이온이 존재할 때 효소의 활성이 나타나는 것을 확인할 수 있었다. 본 연구의 유전자는 낮은 온도의 stress하에서 발현이 증가됨을 Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR)을 통해 확인하였고, 상동성 재조합 (homologous recombination)에 의한 돌연변이 균주 제작을 통해 PMM/PGM protein과 lipopolysaccharide (LPS)의 생합성과의 관계를 규명하였다. V. anguillarum wild type과 mutant로부터 LPS를 분리하였고 sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)후 silver staining을 통해 LPS의 high molecular weight (HMW) 부분인 O-antigen에서의 변화를 확인하였다. 또한 V. anguillarum wild type과 mutant의 growth와 viability를 확인한 결과 mutant가 wild type보다 정지기까지 더 낮은 생육을 보였으며 viability가 감소함을 확인하였다. 본 연구를 통하여 V. anguillarum의 pmm/pgm 유전자가 미생물의 생육과 LPS 생합성에 관여하고 있음을 알 수 있었다.

• Fisheries and Aquatic Sciences
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• 제5권3호
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• pp.178-182
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• 2002

The authors identified 68 Vibrio strains from Gwangan beach seawater from June to October in 2001. We identified them as 19 strains of Vibrio alginolyticus, 15 strains of V. vulnificus, 15 strains of V. parahaemolyticus, 11 strains of V. cholerae non O1, 7 strains of V. fluvialis and just one strain of V. hollisae. They showed their typical biochemical characteristics by API 20E kit (bioMerieux), respectively. It was examined whether their cultural supernatants had enzymatic activities such as hemolysin, protease or urease. The 46 strains showed hemolytic activities and/or protease activities. But we could not find any strain which had urease activity. All isolates of V. cholerae non O1 showed $\beta$ hemolysis. The others showed $\alpha$ hemolysis or did not show clear zones on sheep blood agar plates. These results of Kanagawa phenomenon were not always correspondant with hemolytic activities of cultural supernatants at late log phase. Some strains had higher hemolytic activities despite of showing protease activities on skim milk agar plates and in litmus milk media. On the other hand, some strains showed protease activities but did not show hemolytic activities. Therefore we could guess that there were the relationships between hemolysins and proteases produced by pathogenic vibrios.

• 한국동물위생학회지
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• 제38권2호
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• pp.65-70
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• 2015

The present study was to investigate in vitro antimicrobial effects of propolis against six different fish bacterial pathogens, Aeromonas hydrophila, Edwardsiella tarda, Vibrio vulnificus, V. parahaemolyticus, A. salmonicida subsp. masoucida, A. salmonicida subsp. salmonicida) using minimum inhibitory concentration (MIC) and minimum bacteriocidal concentration (MBC) tests. In the results, propolis exhibited antimicrobial activity against all bacteria used in the present study, but there was no marked difference in bacterial species except Vibrio species. Collectively, propolis was thought to be an usefulness antimicrobial substance for controlling bacterial diseases in the fish industry.

• 한국동물위생학회지
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• 제36권4호
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• pp.297-301
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• 2013

The present study was to investigate anti-microbial effects of curcumin on major bacterial pathogens for farmed fish, such as Aeromonas hydrophila, A. salmonicida subsp. masoucida, A. salmonicida subsp. salmonicida, Edwardsiella tarda, Vibrio vulnificus, V. paraheamolyticus using disk diffusion, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) tests. In disc diffusion test, curcumin exhibited concentration-dependent antimicrobial activities to all bacteria pathogens used in the study. Antimicrobial effects of curcumin was found differently depending on bacterial species when determined by MIC or MBC tests. For examples, E. tarda and A. hydrophila was respectively the most sensitive bacterium for bacteriostatic and bacteriocidal effect of curcumin. Collectively, curcumin could be a potential natural drug for controlling pathogenic bacteria in the aquaculture industry.

• 한국미생물·생명공학회지
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• 제46권2호
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• pp.127-134
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• 2018

본 연구에서는 해조류인 Umbraulva japonica의 표면에서 79개의 세균을 분리하였다. 16s rRNA 유전자 분석 결과, 주요 계통군은 Proteobacteria (74.69%), Actinobacteria (2.53%), Fimicute (2.53%), Bacteroidetes (20.25%)로 4개의 문(Phylum)이 관찰되었고, 7개의 강(Class), 13개의 목(Order), 17개의 과(Family), 31개의 속(Genus)을 확인하였다. 계통학적 분석 결과 3개의 균주가 표준균주와 97% 이하의 유사성을 보여 신속 또는 신종으로 보고될 가능성이 있다고 여겨지며, 향후 표준균주들과 함께 추가적인 신종 실험이 수행되어야 할 것으로 사료된다. 분리된 79 균주를 이용하여 인체 및 어류 병원균을 대상으로 항균 활성을 확인하였다. UJT7, UJT20, UJR17의 균체 현탁액이 Vibrio vulnificus에 대하여 항균 활성을 나타냈으며 UJR17의 균체 현탁액은 V. vulnificus와 Streptococcus parauberis에 항균 활성능이 있음을 확인하였다. UJT7은 Bacillus sp., UJT20과 UJR17은 Pseudomonas sp.로 확인되었으며 다양한 활용을 위한 추가적인 실험을 수행한 후 유익하게 이용 될 수 있을 것이라 사료된다.