• 한국수산과학회지
• /
• 제27권2호
• /
• pp.107-113
• /
• 1994

우리나라 남해연안 어패류에서 V. vulnificus, V. cholerae non O1 균을 분리하여 이들 균이 생산한 용혈독소의 활성을 검토하고 특히 치사율이 높은 패혈증 원인균인 V. vulnificus균이 생산한 균체외 단백독소인 hemolysin을 분리정제하고 얻어진 독소를 이용하여 항혈청을 만들었다. 1. V. vulnificus hemolysin(VVH)은 HI broth에서 $37^{\circ}C,\;15{\sim}24hr$ 진탕배양으로 잘 생산되었으며 V. cholerae non O1 균의 경우는 배양 15시간까지는 hemolysin 생산이 증가되었으나 15시간 경과 후에는 균종에 따라 증가되는 것도 있었고 경과 시간에 따라 오히려 감소하는 균주도 있었다. 2. V. vulnificus가 생산한 hemolysin은 면양적혈구에 대한 용혈활성이 강하고 토끼적혈구에 대하여는 약하였으나 V. cholerae non O1 균주는 토끼적혈구에 대한 용혈활성이 면양이나 말 적혈구에 대한 활성보다 2배정도 강하였다. 3. VVH는 hydrophobic Phenyl-Sepharose HP column을 이용하여 washing buffer와 elution buffer의 성분과 pH를 조정하면서 $1\%$ CHAPS를 이용하여 2차에 걸쳐 column chromatography한 결과 정제도와 수율이 매우 좋아졌다. 본 방법으로 다섯 차례에 걸쳐 정제한 결과 정제된 VVH의 specific activity는 $16900{\sim}52300$배로 평균 27,000배 이상 증가하였으며 수율도 $18.2{\sim}33.0\%$로 평균 $23.4\%$나 되었다. 실제로, V. vulnificus 배양액 2400ml로 부터 정제된 hemolysin을 $250{\mu}g$정도 만들 수 있어서 패혈증 비브리오균 연구에 크게 이바지 할수 있을 것으로 사료된다. 4. 정제된 VVH를 SDS-PAGE한 결과 분자량은 50KDa이었으며 토끼를 이용해서 만든 항혈청의 항체가는 $2000{\sim}8500$이었다.

• 한국수산과학회지
• /
• 제52권1호
• /
• pp.27-34
• /
• 2019

The seasonal and spatial variation of pathogenic Vibrio species, such as V. parahaemolyticus, V. vulnificus, V. alginolyticus, and V. cholerae were investigated in seawater and in bivalves off the Gyeongnam coast of Korea, which is an important area for shellfish production, during the period 2013-2016. V. parahaemolyticus, V. vulnificus, V. alginolyticus, and V. cholerae were detected in 12.1%, 5.2%, 15.4%, and 0.9% of seawater samples, respectively. V. parahaemolyticus, V. vulnificus, V. alginolyticus, and V. cholera were detected in 21.9%, 7.1%, 12.2%, and 0.0% of shellfish samples, respectively. The Vibrio spp. in seawater and bivalve samples were detected at high levels during the summer to early autumn; however, the levels were low during the winter. Therefore, their occurrence was seasonally dependent and correlated with high water temperature, which is also the biggest factor contributing to foodborne outbreaks associated with Vibrio. Relatively high detection rates of the strains were also found in the sea area that was continually exposed to inland wastewater. Our findings show that continuous monitoring is needed to reveal the patterns of occurrence of these pathogens from marine samples collected off the Korean coast, to reduce seafood-borne outbreaks caused by Vibrio.

• Toxicological Research
• /
• 제4권2호
• /
• pp.143-149
• /
• 1988

Cytolysin produced by Vibrio vulnificus ATCC 27562 was partially purified by sequential ammonium sulfate precipitation, gel filtration with Sephadex G-200, and ion exchange chromatography with DEAE-Sephadex. The partially purified cytolysin was inactivated by cholesterol. More than one molecule of the cytolysin was required to lyse a single erythrocyts. The antiserum against cytolysin enhanced the survival ratio of mice infected with low dose of V. vulnificus.

• 한국어병학회지
• /
• 제25권3호
• /
• pp.159-164
• /
• 2012

Vibrio vulnificus ghosts (VVG) were generated using a mobilizable vector including a thermosensitive expression cassette by conjugation. The vaccine potential of VVG was investigated in mouse. Mice immunized with VVG showed significantly higher antibody titer than those with formalin-killed V. vulnificus. The present study supports the conceptive usefulness of bacterial ghosts as vaccine candidates.

• 미생물학회지
• /
• 제50권2호
• /
• pp.108-113
• /
• 2014

녹농균(Pseudomonas aeruginosa)과 비브리오 불니피쿠스균(Vibrio vulnificus)은 그람 음성의 병원균들로써, quorum sensing(QS) 기전을 통해 병원성을 발현하는 세균들이다. 이들 병원균의 감염은 많은 경우 생물막 형성에 의해 매개된다고 알려져 있는데, 이에 본 연구에서는 P. aeruginosa와 V. vulnificus를 대상으로 QS 기전의 유무에 따른 생물막 형성의 시간적 추이를 분석해 보았다. 그 결과 P. aeruginosa의 경우 QS 기전이 결핍된 균주가 야생형에 비해 초기 부착은 더 잘 하였으나, 이후 생물막 구조의 성숙 능력은 야생형에 비해 현저히 떨어짐을 알 수 있었다. 이러한 특성 때문에 야생형과 QS 결핍 균주의 생물막 형성을 시간의 추이에 따라 정량적으로 비교해 보면 초기 10시간 정도 까지는 QS 결핍 균주가 더 많은 생물막을 형성하다가, 이후 야생형이 더 많이 생물막을 형성하는 역전 현상이 관찰되었다. V. vulnificus는 P. aeruginosa와는 달리 QS 결핍 균주가 야생형보다 더 많은 생물막을 형성한다고 보고된 균주이다. 이 균주에서 같은 방식으로 생물막 형성을 조사해 본 결과, 108시간의 장시간 동안에도 항상 QS 결핍 균주가 야생형 보다 더 많은 생물막을 형성하여, 역전 현상은 관찰되지 않았다. 이 결과는 P. aeruginosa의 경우에는 QS 기전이 초기 부착은 저해하는 방향으로, 성숙과정은 촉진시키는 방향으로 작용하며, V. vulnificus에서는 일관되게 생물막 형성을 저해하는 방향으로 작용함을 보여주는 것이다. 따라서 생물막 제어를 위한 타겟으로 QS기전을 이용할 때에는 제어하고자 하는 생물막 형성 단계와 세균 종을 함께 고려하여야 한다고 제안한다.

• Fisheries and Aquatic Sciences
• /
• 제24권6호
• /
• pp.207-218
• /
• 2021

This study evaluated the risk of foodborne illness from highly pathogenic Vibrio spp. (Vibrio vulnificus and V. cholerae) by raw whip-arm octopus (Octopus minor) consumption. In total 180 samples of raw whip-arm octopus were collected from markets and examined for the prevalence of V. vulnificus and V. cholerae. Predictive models describing the kinetic behavior of Vibrio spp. in raw whip-arm octopus were developed, and the data on amounts and frequency of raw whip-arm octopus consumption were collected. Using the collected data, a risk assessment simulation was conducted to estimate the probability of foodborne illness raw whip-arm octopus consumption using @RISK. Initial contamination levels of Vibrio spp. in raw whip-arm octopus were -3.9 Log colony-forming unit/g, as estimated by beta distribution fitting. The developed predictive models were appropriate to describe Vibrio spp. in raw whip-arm octopus during distribution and storage with R² values of 0.946-0.964. The consumption frequency and daily consumption amounts of raw whip-arm octopus per person were 0.47% and 57.65 g, respectively. The probability of foodborne illness from raw whip-arm octopus consumption was estimated to be 8.71 × 10^-15 for V. vulnificus and 7.08 × 10^-13 for V. cholerae. These results suggest that the risk of Vibrio spp. infection from raw whip-arm octopus consumption is low in South Korea.

• Journal of Microbiology
• /
• 제45권2호
• /
• pp.146-152
• /
• 2007

A mutant exhibiting decreased cytotoxic activity toward INT-407 intestinal epithelial cells and carrying a mutation in the rtx gene cluster that consists of rtxCA and rtxBDE operons was screened from a library of V. vulnificus mutants. The functions of the rtxA gene, assessed by constructing an isogenic mutant and evaluating its phenotypic changes, demonstrated that RtxA is essential for the virulence of V. vulnificus in mice as well as in tissue cultures.

• Journal of Microbiology and Biotechnology
• /
• 제16권5호
• /
• pp.808-816
• /
• 2006

A wbpO gene encoding a putative UDP-N-acetylo-galactosamine dehydrogenase was identified and cloned from Vibrio vulnificus. LPS production was altered by disruption of the wbpO gene through allelic exchanges. The function of the wbpO gene in virulence assessed in vitro and in mice revealed that WbpO is important in both the pathogenesis of V. vulnificus and the biosynthesis of LPS.

• Journal of Life Science
• /
• 제9권1호
• /
• pp.35-39
• /
• 1999

Elecrophoreticl analysis of a 36 kDa protein was runned by SDS-PAGE, isoelectric focusing (IEF) and two dimensional electrophoresis pattern. Major 36 kDa and 25, 46, 48, 66 kDa protein were detected by Coomassie blue stain on SDS-PAGE. Major 36kDa protein was eluted for production of antiserum for serological analysis, IEF and two dimensional electrophoresis. Isoelectric point of 36kDa was aout pH 8.5. Two dimensional electrophoresis of eluted 36kDa showed one point on the gel. Anti-36 kDa serum made by newzilland rabbit for serological test. In ELISA, final titer of antibody was 100×{TEX}$2^5}${/TEX} : 1. Neutralize ability of serum was examined by slide agglutination test and colonization test in rat. Anti-36 kDa serum agglutinated whole cell of V. vulnificus were inhibited colonization on intestine in rat. Accordingly In this paper contain some electrophoretical analysis and serological test of a 36 kDa OMP of V. vulnificus.

• Journal of Microbiology and Biotechnology
• /
• 제18권5호
• /
• pp.968-974
• /
• 2008

Proteomic analysis led to identification of the proteins of Vibrio vulnificus that were induced upon exposure to INT-407 cells, and 7 of which belong to the functional categories such as amino acid transport/metabolism, nucleotide transport/metabolism, posttranslational modification/protein turnover/chaperones, and translation. Among the genes encoding the host-induced proteins, disruption of purH, trpD, tsaA, and groEL2 resulted in reduced cytotoxicity. The purH, trpD, and tsuA mutants showed impaired growth in the INT-407 lysate; however, the growth rate of the groEL2 mutant was not significantly changed, indicating that the possible roles of the host-induced proteins in the virulence of V. vulnificus are rather versatile.