2018~2022년 경기 해안지역에서 분리된 비브리오패혈증균의 월별 분포 특성

윤수정,편희수,이윤희,박소정,강경자,허은선,정일형,김범호,권순목 한국환경보건학회 2024 한국환경보건학회지 Vol.50 No.1

Background: Vibrio vulnificus is a serious opportunistic human pathogen that has a worldwide distribution in a variety of marine and estuarine environments. Objectives: For this reason, we investigated the distribution of Vibrio vulnificus in coastal areas of Gyeonggido Province from 2018 to 2022. Also, we analyzed the correlation between V. vulnificus leading to infection and two marine environmental factors (water temperature and salinity). Methods: We collected a total of 266 samples from six coastal area points (i.e., seawater, mudflats). Specimens were isolated using selective plating media and isolated strains were identified by a VITEK 2 system. To find the relevance of the isolation rates of V. vulnificus and number of cases of V. vulnificus infection, we summarized the data on 48 cases of V. vulnificus infection from the open data of the Korea Disease Control and Prevention Agency. Results: Among the 266 samples taken during the investigation period, 47 strains were isolated, and the separation rates of V. vulnificus were 17.7%. The monthly isolation rates of V. vulnificus were ranked in the order of August (53.8%), September (33.3%), June (28.6%), and July (21.1%). There was a positive correlation with the temperature of seawater, but salinity was not significant. The number of cases of V. vulnificus infection reported in Gyeonggi-do Province were 18 (37.5%) in September, 14 (29.2%) in August, and eight (16.7%) in October. The proportion was 83.3%. It was relevant to the isolation rates of V. vulnificus in the marine environmental sources. Conclusions: Our data showed that the number of V. vulnificus infection cases could be affected by changes in the distribution of V. vulnificus due to rise the temperature of seawater in the marine environment.

The In Vitro and In Vivo Efficacy of Hen IgY Against Vibrio parahaemolyticus and Vibrio vulnificus

( Kassim Neema ),( Adelard B Mtenga ),( Won Bo Shim ),( Duck Hwa Chung ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.10

The inhibitory effect of IgY against Vibrio parahaemolyticus and Vibrio vulnificus responsible for seafood-borne diseases was investigated in this study. Water-soluble fractions (WSF) of protein containing IgYs were isolated from the egg yolk of hens initially immunized with formalininactivated V. parahaemolyticus or V. vulnificus. Protein, total and specific IgY contents of the WSF were determined. The inhibitory and protective effects of IgYs on the growth of V. parahaemolyticus and V. vulnificus were assayed in liquid medium and in mice. IgYs showed high affinity to their corresponding antigens with high titer from day 28 onwards. Protein contents and total IgY concentrations remained stable throughout the immunization period, whereas specific IgY concentrations increased steadily and reached a plateau at day 49. Specific IgY powder (150 mg/ml) significantly inhibited further multiplication of both V. parahaemolyticus and V. vulnificus in liquid medium as compared with the control IgY. The bacteria count in mice feces was lower in mice pretreated with specific IgYs than in those pretreated with PBS or control IgY. Higher survival of mice was observed in the experimental groups pretreated with either anti-V. parahaemolyticus (75% survival) or anti-V. vulnificus (87% survival) IgYs, compared with those in the control groups pretreated with PBS or nonspecific IgY. All mice in the control groups died within three days after bacteria inoculation; hence, the protective effect of specific IgYs against infection caused by V. parahaemolyticus and V. vulnificus was demonstrated.

V. vulnificus의 cyolysin에 의해 형성된 Pored의 이온 투과성에 관한 연구

마천택 ( Tian Ze Ma ),홍병철 ( Bing Zhe Hong ),고경희 ( Hee Ko Kyeung ) 전북대학교 의과학연구소 2004 全北醫大論文集 Vol.28 No.1

V. vulnificus가 분비하는 cytolysin의 pore 형성 효과를 전기생리학적으로 확인하고 그 pore의 이온선택성을 밝히고자, V. vulnificus cytolysin의 표적세포로 예산되는 폐내피세포인 CPAE 세포에서 여러 bath용액에서의 cytolysin pore의 이온투과성을 비교 관찰하여 다음과 같은 결과를 얻었다. 1. Cytolysin 1 HU/ml을 세포내외에 투여시 2-7분의 lag time 후 sensitivity 차이없이 115.6 ± 5.1 pS의 단일 전도도를 갖는 pore를 시간 의존적으로 형성하였으며, 이런 pore의 이온투과성은 세척 후에도 지속적으로 유지되었다. 2. Bath용액의 140 mM KCI을 같은 농도의 NaC1 및 CsC1로 치환하여도 cytolysin pore 의 이온투과성에는 유의한 변동이 없었다. 3. Bath용액의 140 mM KCI을 같은 농도의 CaC1(2) 및 BaCl2로 치환하면 cytolysin pore의 이온투과성이 각각 1.8배와 2.3배로 현저하게 증가하였다. 4. Bath용액의 C1 농도를 동일하게 하고자, 각각 140 mM KCI과 70 mM MgC1(2)로 치환시 cytolysin pore의 이온투과성에는 유의한 차이가 없었다. 5. Bath용액의 KCI농도를 140 mM에서 10mM과 1 Mm로 감소시 cytolysin pore의 역전전압이 0 mV에서 각각 -31.1 ± 1.5mV와 -42.3 ± 1.8 mV로, C1 이온의 역전전압에 근접하게 이동하였다. 이상의 실험결과를 종합하변, V. vulnificus cytolysin은 세포막내외의 차이없이 시간의존 적인 pore를 형성하는데, 이 pore의 투과성은 막전위에 비의존적이며 주로 C1 이온에 선택성이 높음을 시사한다. Cytolysin produced by V. vulnificus has been incriminated as one of the important virulence determinants in V. vulnificus infection. Ion selectivity of cytolysin-induced pres has been examined in CPAE cells, a cell line of pulmonary endothelial cells, using inside-out patch clamp techniques. In symmetrical KCI concentration (140 mM). intracellular and extracellular cytolysin formed the ion-permeable pores with single channel conductance of 115.6±5.1 pS (n=6) in a time-dependent manner and a voltage-independent manner. The pore currents were consistently maintained after washout of cytolysin. Replacement of intracellular 140 mM KCI with 140 mM NaCl, 140 mM CsCl and 70 mM MgCl2 did not affect the pore currents, but replacement of it with equimolar CaCl2 and BaCl2 increased the pore current by 1.8 and 2.4 times of control current (140 mM KCL), respectively. When the intracellular KCI concentration was lowered from 140 to 10 and 1 mM, zero-current membrane potentials shifted from 0 mV to -31.1 ± 1.5 and -42.3 ± 1.8 mV, respectively. These results indicate that cytolysin produced by V. vulnificus formed the CI(-) -selective pores in CPAE cells.

Bacteriocin ("Vulnificin") Typing of Vibrio vulnificus

Ha, Tai-You,Lee, Jeong-Ho,Lee, Hern-Ku,Whang, Hee-Sung,Park, Jae-Seung The Korea Society for Microbiology 2000 大韓微生物學會誌 Vol.35 No.3

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.

Genistein Inhibits Vibrio vulnificus Adhesion and Cytotoxicity to HeLa Cells

오둘리,Jong Ro Kim,Young Ran Kim 대한약학회 2010 Archives of Pharmacal Research Vol.33 No.5

Vibrio vulnificus (V. vulnificus) is a useful model for bacterial septicemia as the bacterial infection generates a wide pathogenic spectrum in addition to a high mortality rate. This study was aimed at investigating the effects of genistein on the growth, cytotoxicity, adhesion, and the mouse mortality caused by V. vulnificus. The results of our study indicated that genistein (50 or 100 mg/L) effectively minimized the morphologic damages and inhibited acute death of HeLa cells by V. vulnificus. Strikingly, genistein significantly inhibited the adhesion of V. vulnificus to HeLa cells. This report confirmed that genistein showed bacteriostatic activity against V. vulnificus, but it did not exhibit any bactericidal activity. Nevertheless, genistein (20 mg/kg) effectively decreased CD-1 mice mortality caused by V. vulnificus infection.

Abundance, antimicrobial resistance, and virulence of pathogenic <i>Vibrio</i> strains from molluscan shellfish farms along the Korean coast

Mok, Jong Soo,Ryu, Ara,Kwon, Ji Young,Park, Kunbawui,Shim, Kil Bo Elsevier 2019 MARINE POLLUTION BULLETIN Vol.149 No.-

<P><B>Abstract</B></P> <P>To reduce the outbreaks caused by the major pathogenic <I>Vibrio</I> species, <I>V. parahaemolyticus</I>, <I>V. vulnificus</I>, and <I>V. cholerae</I>, the distribution, antibiotic resistance, and virulence of these <I>Vibrio</I> strains were monitored in shellfish and seawater along the Korean coast. Among the <I>Vibrio</I> strains, <I>V. parahaemolyticus</I> was the most abundant species; during summer, this strain showed a substantial increase that correlated with the water temperature. Although >99.0% of the <I>Vibrio</I> species isolates were sensitive to seven antimicrobials recommended by the Center for Disease Control and Prevention for the treatment of <I>Vibrio</I> infections, multiple-antibiotic resistance to at least three antimicrobials was found in 14.3% to 50.0% of each <I>Vibrio</I> species. Among <I>V. parahaemolyticus</I> isolates, 14.3% were positive for the <I>trh</I> gene, whereas only 1% was positive for the <I>tdh</I> gene. These results should aid in implementing proper precautions to avoid potential human health risks associated with exposure to pathogenic <I>Vibrio</I> species.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The major <I>Vibrio</I> species are <I>V. parahaemolyticus</I>, <I>V. vulnificus</I>, and <I>V. cholerae</I>. </LI> <LI> <I>V. parahaemolyticus</I> was the most abundant strain among the pathogenic <I>Vibrio</I> species. </LI> <LI> More than 99% of <I>Vibrio</I> isolates were sensitive to 7 antibiotics suggested by the CDC. </LI> <LI> The 14.3% to 50.0% of each <I>Vibrio</I> species was resistant to at least three antibiotics. </LI> <LI> Virulent <I>V. parahaemolyticus</I> accounted for 14.8% of the isolates </LI> </UL> </P>

금강 하구 해역의 해수에 병원성 비브리오균(Vibrio spp.)의 분포

박선아,박권삼 한국수산과학회 2022 한국수산과학회지 Vol.55 No.6

The pathogenic Vibrio genus denotes halophilic bacteria that are distributed in aquatic environments, including both sea and freshwater. V. cholerae, V. vulnificus, and V. parahaemolyticus are the main species that can be potent human pathogens and the leading cause of septicemia, wound infections, and seafood borne gastroenteritis. The aim of this study was to investigate the presence of pathogenic Vibrios in seawater. We obtained a total of 80 seawater samples from the Geum River estuary area in the west coast of Korea from April to December 2021. Pathogenic Vibrios was determined using a combination of the most probable number-polymerase chain reaction (MPN-PCR) methods. The detection levels of V. cholerae, V. parahaemolyticus, and V. vulnificus in the seawater samples were 7.5%, 68.8%, and 30.0%, respectively. The quantitative results were as follows: 3.6–3.6 MPN/100 mL in V. cholerae, 3.6–3,400 MPN/100 mL in V. parahaemolyticus, and 3.6–4,300 MPN/100 mL in V. vulnificus. Overall, these results provide novel insight into the necessity for seawater sanitation in the Geum River estuary area, and could help reduce the risk of seafood-borne outbreaks caused by pathogenic Vibrios.

인천지역에서 분리된 비브리오 패혈증균의 특성

오보영,김정희,공용우,제갈승,김혜영,이미연,황경화,고연자,이제만,고종명,김용희,Oh, Bo-Young,Kim, Jung-Hee,Gong, Young-Woo,JeGal, Seung,Kim, Hye-Yeung,Lee, Mi-Yeon,Hwang, Kyoung-Wha,Koh, Yeon-Ja,Lee, Jae-Mann,Go, Jong-Myoung,Kim, Yong-H 한국미생물학회 2007 미생물학회지 Vol.43 No.4

2006년도 인천지역 해양환경에서 분리된 Vibrio vulnificus에 대한 생화학적, 분자생물학적 특성 및 항생제 감수성 결과를 알아보았다. 본 실험에 사용된 균주는 총 233주로 해수, 갯벌, 어패류, 수족관수에서 분리되었다. API 20E kit 실험 결과 15개 profile로 분류되었으며, 모든 균주가 ONPG와 Amygdalin 양성이었다. 209주를 대상으로 vvhA와 viuB 유전자 부위에 대해 PCR 실험결과 vvhA는 206주(98.6%) 양성, viuB는 110주(52.6%)가 양성이었으며, 특히 viuB 유전자부위에 대한 PCR 결과는 해수, 어패류, 갯벌에서 분리한 균주의 48%, 48.5%, 61.1%가 양성인 것으로 나타났다. 시험균주 175주에 대해 항생제 감수성 실험 결과 Imipenem (100.0%), Sulfamethoxazole/trimethoprim (98.9%), Tetracycline, Ciprofloxacin (98.3%), Ampicillin/sulbactam (97.1%), Chloramphenicol (96.6%), Cefepime (94.9%), Ceftriaxone (94.8%)이 감수성을 나타내었고, 항생제 하나 또는 그 이상의 약제에 대해 내성을 나타낸 것은 56주(32%)로 해수 28주(31.5%), 갯벌 21주(34.4%), 어패류 7주(29.2%)였다. V. vulnificus 233주에 대해 PFGE를 실시하여 dendrogram으로 분석한 결과 90%이상의 상동성 기준으로 126개의 유형으로 분리되었고, 58%이상의 상동성을 기준으로 13개의 cluster로 분류되었다. Cluster I는 104주(44.6%)로 가장 많은 균주들이 분포하였고 채취시기 대부분 I에 가장 많은 균주들이 분포하였으나 10월과 6월에 채취한 검체는 J에 16주(69.6%)와 13주(36.1%)로 가장 많은 균주들이 분포하였다. 7월에 채취한 검체에서 분리된 균주는 9개의 cluster에 속하였고 8월은 8개, 6월은 7개, 9월은 6개, 10월은 5개, 5월은 3개, 3월은 1개를 나타냈다. We performed the biochemical characteristics, molecular epidemiologocal analysis, and drug susceptibility test on V. vulnificus isolated from environmental sources in Incheon. For this study, 233 strains were isolated from seawater, sediment, shellfish. V. vulnificus isolates were divided into 15 biochemical groups, which were positive for ONPG and Amygdalin test. Among the 209 strains, 206 (98.6%) strains and 110 (52.6%) strains revealed positive for vvhA and viuB gene, and the viuB gene detection rates of V. vulnificus from seawater, shellfish and sediment were 48%, 48.5% and 61.6%, respectively. From disc diffusion test on 175 isolates, most of strains were sensitive to Imipenem (100.0%), Sulfamethoxazole/trimethoprim (98.9%), Tetracycline, Ciprofloxacin (98.3%), Ampicillin/sulbactam (97.1%), Ohloramphenicol (96.6%), Cefepime (94.9%) and Ceftriaxone (94.8%), multi-drug resistance rates was 31.5% of seawater, 34.4% of sediment and 29.2% of shellfish. PFGE was performed on 233 V. vulnificus isolates with the objective of investigating the extent of genetic diversity of these isolates in our region. We could find that at least 126 different PFGE patterns were generated according by 90% of similarity and 13 clusters by 58% of similarity. The major cluster was type I (44.6%) during the most of the year, and type J was frequent pattern in June and October. There were 9 distinct PFGE types in July, 8 types in August, 7 types in June, 6 types in September, 5 types in October 3 types in May and 1 type in March.

Characterization of Prohibitin 1 as a Host Partner of <i>Vibrio vulnificus</i> RtxA1 Toxin

Kim, Bo A.,Lim, Ju Young,Rhee, Joon Haeng,Kim, Young Ran Oxford University Press 2016 The Journal of Infectious Diseases Vol.213 No.1

<P>RtxA1 toxin, which results in cytoskeletal rearrangement, contact cytotoxicity, hemolysis, tissue invasion, and lethality in mice, is the most potent cytotoxic virulence factor of Vibrio vulnificus. Bioinformatics analysis of rtxA1 predicted 4 functional domains that presumably performed discrete functions during host cell killing. V. vulnificus RtxA1 has a unique domain designated as RtxA1-D2, corresponding to amino acids 1951-2574, which is absent in Vibrio cholerae multifunctional-autoprocessing repeats-in-toxin, suggesting that this domain confers specific biological functions to V. vulnificus RtxA1. HeLa cells expressing green fluorescent protein-RtxA1-D2 became round and lost their viability. A yeast 2-hybrid system identified prohibitin (PHB) 1 as the host partner of RtxA1-D2. The specific interaction of RtxA1-D2 with PHB1 was confirmed by performing immunoprecipitation. Interestingly, V. vulnificus RtxA1 up-regulated PHB1 expression on the cytoplasmic membrane of host cells. Extracellular signal-regulated kinase and p38 mitogen-activated protein kinase pathways were confirmed as being important in the up-regulation of PHB1 by using inhibitors. Down-regulation of PHB1 by small interfering RNAs decreased the cytotoxicity of RtxA1-D2 against HeLa cells. The pretreatment of an anti-PHB1 antibody impaired the cytotoxicity of V. vulnificus RtxA1. These results suggest that the involvement PHB1 in the RtxA1 cytotoxicity has significant implications for the pathogenesis of V. vulnificus infections.</P>

Characterization of Prohibitin 1 as a Host Partner of Vibrio vulnificus RtxA1 Toxin

( Bo A. Kim ),( Ju Young Lim ),( Joon Haeng Rhee ),( Young Ran Kim ) 전남대학교 약품개발연구소 2015 약품개발연구지 Vol.24 No.-

RtxA1 toxin, which results in cytoskeletal rearrangement, contact cytotoxicity, hemolysis, tissue invasion, and lethality in mice, is the most potent cytotoxic virulence factor of Vibrio vulnificus. Bioinformatics analysis of rtxA1 predicted 4 functional domains that presumably performed discrete functions during host cell killing. V. vulnificus RtxA1 has a unique domain designated as RtxA1-D2, corresponding to amino acids 1951-2574, which is absent in Vibrio cholerae multifunctional-autoprocessing repeats-in-toxin, suggesting that this domain confers specific biological functions to V. vulnificus RtxA1. HeLa cells expressing green fluorescent protein-RtxA1-D2 became round and lost their viability. A yeast 2-hybrid system identified prohibitin (PHB) 1 as the host partner of RtxAI-D2. The specific interaction of RtxAI-D2 with PHBI was confirmed by performing immunoprecipitation. Interestingly, V. vulnificus RtxA1 up-regulated PHBI expression on the cytoplasmic membrane of host cells. Extracellular Signal-regulated kinase and p38 mitogen-activated protein kinase pathways were confirmed as being important in the up-regulation of PHBI by using inhibitors. Down-regulation of PHBI by small interfering RNAs decreased the cytotoxicity of RtxAI-D2 against HeLa cells. The pretreatment of an anti-PHBI antibody impaired the cytotoxicity of V. vulnificus RtxA1. These results suggest that the involvement PHBI in the RtxA1 cytotoxicity has Significant implications for the pathogenesis of V. vulnificus infections.