• 대한미생물학회지
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• 제35권3호
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• pp.225-237
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• 2000

Vibrio vulnificus, a halophilic vibrio is an estuarine gram-negative bacteria that is associated with severe and frequently fatal wound infections and life-threatening septicemia. Bacteriocins are defined as antibacterial substance produced by various species of bacteria which are usually active against closely related organisms. Bacteriocins have found widespread application in epidemiological studies as specific markers of bacteria. It was proposed by Ha et al. (1990. J. Korean. Soc. Microbiol. 25: 586.) to give the bacteriocins produced by V. vulnificus the name "vulnificins". In the present study, a total of 72 strains of V. vulnificus isolated from patients and oysters were subjected to screen potential producers and indicators of vulnificin, applying ultraviolet induction method. Sensitivity of several strains of Serratia marcesans, Pseudomonas aeruginosa, Shigella flexneri, Salmonella typhi and Yersinia enterocolitica to vulnificins were also examined out. All the tested strains of V. vulnificus produced vulnificins active against indicator strains with various different inhibitory patterns. The spectrum of vulnificin activity and sensitive spectrum of indicator strains were considerably broad. Interestingly, almost all strains of S. marcescens, P. aeruginosa, Salmonella sp., Shigella sp. and Y. enterocolitica tested were sensitive to 1-7 vulnificin(s). Taken together, the present study demonstrated that all of the isolates of V. vulnificus produced vulnificins and that 8 good vulnificin producers and 10 good indicators were detected. These strains can be employed efficiently for establishing vulnificin typing scheme of V. vulnificus and for the detection of bacteriocinogeny and sensitivity in V. vulnificus. Biological role of vulnificin remains to be further elucidated.

• 대한미생물학회지
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• 제22권2호
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• pp.109-116
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• 1987

As a process to establish an effective preventive measure of V. vulnificus septicemia, bactericidal effect of distilled water against V. vulnificus was studied. When about $2.0{\times}10^7\;CFU/ml$ of V. vulnificus was inoculated in distilled water, a dramatic decrease in the number of viable bacteria by 5 to $6LOG_{10}$ was observed in 5 minutes. Bactericidal kinetic curves could be divided into the first rapid killing phase until 1 minute and the later slow killing phase after then, showing the heterogeneity of the bacterial population inoculated. When V. vulnificus was inoculated in saline solutions having various salinities, significant decrease in the number of viable bacteria was noted only at salinities under 0.2%. The higher was the concentration of NaCl, the greater was the degree of protection against osmotic shock. When glucose, NaCl, $MgCl_2$, and $CaCl_2$ were diluted with deionized water to give same osmolarities and V. vulnificus was inoculated in each of them to compare the bactericidal curves plotted during the first 5 minutes after inoculation, the protection efficiencies were in the order of $MgCl_2>CaCl_2{\gg}NaCl{\gg}glucose$. Above results indicate that treatment(or thorough washing) of contaminated sea animals or other products with distilled water can be used as a preventive measure of V. vulnificus septicemia, and divalent cations can protect V. vulnifcus to osmotic shock with high efficiency.

• Journal of Life Science
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• 제11권1호
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• pp.27-29
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• 2001

Vibrio vulnificus endotoxin was extracted, analyzed the chemical composition, tested its biological activity, and compared to those of Escherichia coli and Salmonella typhimurium. The major fatty acid of three endotoxins were different each other; V. vulnificus endotoxins were different each other; V. vulnificus endotoxin was myristric acid (C14:0), E. coli was lauric acid (C12:0), S. typhimurium was capric acid (C10:0). The biological activities of V. vulnificus endotoxin were similar to those of E. coli and S. typhimurium in terms of the gelation activity of the Limulus amebocyte lysate and the lethal toxicity. But the result of enzyme (AST, ALT, and LDH) analysis showed that the enzyme activity of V. vulnificus endotoxin was similar to that of E. coli, but smaller than that of S. typimurium.

• Journal of Microbiology and Biotechnology
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• 제11권2호
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• pp.302-309
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• 2001

Colony blot hybridization using a VVHP DNA probe derived from the sequence of the hemolysin gene, vvhA, was specific in identifying all V. vulnificus strains, thereby, eliminating the need for any additional phenotypic identification. The colony blot hybridization procedure revealed a sensitivity and broad applicability sufficient for the direct enumeration of V. vulnificus in various natural samples, without the use of enrichment or culturing on selective medis. V. vulnificus was detected in all natural samples collected during August and May at concentrations ranging from $2.1{\times}10^1\;to\;4.0{\times}10^3$ organisms per ml. However, during November and February, when the mean temperatures of the seawater were $12^{\circ}C$ and $5^{\circ}C$, respectively, V. vulnificus was not detected in any natural samples.

• Journal of Microbiology
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• 제35권2호
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• pp.87-91
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• 1997

Monoclonal antibodies were prepared in order to an assay method for Vibrio vulnificus. Sixteen mouse ybridoma cell lines were established by immunization of whole cell antigen to BALB/c mice, fusion with SP2/O myeloma cells, and cloning. Most of them secreted IgM.lambda. antibodies. A sandwich enzyme-linked immunosorbent assay was developed with rabbit anti-V. vulnificus polyclonal antibodies as capture antibody, an IgM monoclonal antibody as detector antibody, and goat anti-mouse IgM-alkaline phosphatase conjugate as developer antibody. The range of detection was 10$\^$4/ to 10 V. vulnificus cells per microplate well. When four related Vibrio species were tested for cross-reactions, V. parahaemolyticus showed 3.5% reactively and V. carchariae, V. fluvialis, and V. furnisii showed negligibal (<1%) cross-reactivity.

• 한국수산과학회지
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• 제19권2호
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• pp.118-126
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• 1986

1985년 7월에서 10월 사이에 부산연안지역에서 수집한 해수, 낙지, 피조개, 게, 우렁쉥이 등 70종의 시료에서 Vibrio vulnificus의 검출률과 시료에서 분리된 균의 세균학적 특성을 조사한 결과는 다음과 같다. 1. Vibrio vulnificus의 월별 검출률은 수온이 높은 8월에 $19.2\%$로 가장 높았으며 7월에 $7.7\%$ 10월에 $5.5\%$로 수온과 비례하여 나타났으며 7월과 10月 사이의 평균검출률은 $11.4\%$였다. 시료별 검출률은 서식해역의 오염도가 높고 오염된 해수와의 접촉부위가 많은 피조개와 게에서 $20\%$로 가장 높았으며 낙지 해수의 순으로 나타났다. 2. 시료에서 분리된 균과 환자에게 분리된 균의 생화학적인 차이점은 없었으며, 분리된 본균이 다른 Vibrio균과 구별될 수 있는 생화학적 특징은 ONPG 가수분해양성, lactose 분해양성, 식염내성이 $7\%$이하인 점이다. 3. 이균의 최적배양조건은 pH 8.0, 온도 $35^{\circ}C$부근이었으며, 배양온도별 비증식속도와 평균세대시간은 $35^{\circ}C$진탕배양에서 $1.21\;hr^{-1}$, 34min으로 가장 빨랐으며 $35^{\circ}C$ 정치배양, $40^{\circ}C$ 정치배양, $25^{\circ}C$ 진탕배양 $25^{\circ}C$ 정치배양의 순이었다. 4. 이 균은 장내세균분리용배지인 EMB, SS, DC 한천배지에서는 증식하지 못했으며 MC 한천배지상에서의 집낙의 특징은 적색 혹은 무색으로 나타났고 Endo 한천배지상에서는 적색, TCBS 한천배지상에서는 녹색집낙으로 나타났다. 5. 이 균을 $4^{\circ}C$에 냉장하였을 경우 매 16시간마다 약 1 log cycle씩 감소하였으며 $-18^{\circ}C$에 동결하였을 때는 거의 다 사멸하였다.

• 한국수산과학회지
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• 제56권5호
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• pp.626-633
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• 2023

Vibrio spp. are aquatic bacteria that are ubiquitous in warm estuarine and marine environments. Especially, V. vulnificus and V. cholerae are currently known to cause potentially fatal infections in humans. This study investigated the distribution and antibiotic resistance of V. vulnificus and V. cholerae isolated from coastal areas of Gyeongsangnam-do in 2022. A total of 252 samples of water, shellfish and coastal sediment were collected from 7 locations along the coast, and 124 samples of fishery products were collected from markets. Among the 252 samples, forty-four V. vulnificus (11.7%) and fourteen V. cholerae non-O1/non-O139 (3.7%), none of which carried the ctx gene, were isolated. Out of the 124 samples, 6 (4.8%) tested positive for V. vulnificus and V. cholerae was not detected. The isolation rates of V. vulnificus and V. cholerae showed a significant correlation with environmental factors such as seawater temperature and salinity. In an antibiotic resistance test, V. vulnificus was susceptible to amikacin, gentamicin, imipenem trimethoprim/sulfamethoxazole, and ciprofloxacin, but resistant to cefoxitin (100.0%), followed by tetracycline (9.1%). Multidrug resistance was also observed. Continuous monitoring of Vibrio pathogens with water temperature and salinity is expected to help reduce the outbreaks, and rational use of antibiotic agents is needed to prevent the accession of antibiotic-resistant microorganisms in aquatic ecosystems.

• 한국식품위생안전성학회지
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• 제33권5호
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• pp.412-415
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• 2018

V. vulnificus는 그람음성의 호염성균으로 감염 되었을 경우, 복통과 발열 등의 급성 위장염을 일으키며 만성질환자에게 급성 패혈증을 일으키는 매우 높은 치사율의 식중독균이다. 식품 중에서 V. vulnificus를 분석하는 방법으로는 TCBS agar와 같은 선택배지를 이용하는 방법과 PCR을 이용한 방법이 있으나 온도, 염 및 pH 등과 같은 환경 요인에 민감한 V. vulnificus의 특성을 고려하였을 때 정확한 균수 정량을 위한 정량분석법 확립의 필요성이 요구된다. 본 연구에서는 배지 및 염 차이에 따른 V. vulnificus 생육 특성 차이에 대한 연구를 진행하였다. 그 결과, V. vulnificus 균수 정량분석에 APW 증균 배양을 이용한 MPN-PCR 방법이 적합하였다. 본 연구에서 제시된 방법은 해수뿐만 아니라 어패류 등의 시료에서 V. vulnificus 균수 정량분석에 유용하게 사용될 것으로 기대된다.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.350-355
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• 2004

Sulforhodamine B (SRB) assay is a rapid, sensitive, and inexpensive method for measuring cell proliferation and chemosensitivity. However, the lactate dehydrogenase (LDH) release assay is generally used to measure cytototoxicity of infectious microorganisms against host cells. In this study, we investigated the possibility of applying the SRB assay to determine cytotoxicity for infectious microorganisms, and compared the results with those obtained by the LDH release assay. We used Vibrio vulnificus as a model of infectious microorganisms. The SRB assay showed that V vulnificus strongly induced cytotoxic activity against human intestinal cells, Caco-2 and INT-407 cells. The degree of cytotoxicity closely correlated with infection time and number ratios of V. vulnificus to intestinal cells (MOI, multiplicity of infection). Furthermore, cytotoxicity values obtained by SRB assay correlated well with results obtained by the LDH release assay, and both assays gave a linear response with respect to MOI Heat-inactivation of V. vulnificus for 35 min at $60^{\circ}C$ did not induce cytotoxic activity, indicating that viability of V. vulnificus is crucial for cytotoxic activity against intestinal cells. Although both assays are suitable as cytotoxicity endpoints, the SRB assay is recommended for measuring cytotoxicity of infectious microorganisms against host cells because of its significantly lower cost and more stable endpoint than the LDH release assay.

• Journal of Microbiology
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• 제43권spc1호
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• pp.118-131
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• 2005

Vibrio vulnificus is an opportunistic pathogen of humans that has the capability of causing rare, yet devastating disease. The bacteria are naturally present in estuarine environments and frequently contaminate seafoods. Within days of consuming uncooked, contaminated seafood, predisposed individuals can succumb to sepsis. Additionally, in otherwise healthy people, V. vulnificus causes wound infection that can require amputation or lead to sepsis. These diseases share the characteristics that the bacteria multiply extremely rapidly in host tissues and cause extensive damage. Despite the analysis of virulence for over 20 years using a combination of animal and cell culture models, surprisingly little is known about the mechanisms by which V. vulnificus causes disease. This is in part because of differences observed using animal models that involve infection with bacteria versus injection of toxins. However, the increasing use of genetic analysis coupled with detailed animal models is revealing new insight into the pathogenesis of V. vulnificus disease.