• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1122-1126
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• 2009

The exogenously added cadaverine is effective in protecting Vibrio vulnificus from methyl viologen (MV)-induced superoxide stress at pH 8.5. Such a protective effect by cadaverine was not observed at pH 7.5. Consistently, the accumulated level of intracellular cadaverine at pH 8.5 is approximately four times as much as that of the control cell at pH 7.5. Cadaverine accumulation is not affected by MV. The protection of V. vulnificus by cadaverine from superoxide stress was abolished when cadB coding for the lysine-cadaverine antiporter was interrupted. However, the cadaverine-mediated protection was complemented with cadB DNA. Therefore, CadB of V. vulnificus not only acts as a lysine-cadaverine antiporter at acid pH to neutralize the external medium, but also mediates cadaverine uptake at alkaline pH to result in cell protection from superoxide stress.

• Journal of Microbiology
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• v.42 no.2
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• pp.69-73
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• 2004

Vibrio vulnificus, a Gram-negative bacterium found in estuarine waters, is responsible for over 95％ of all seafood-related deaths in the United States. As a result of a temperature downshift to 5$^{\circ}C$, this organism enters the viable but nonculturable (VBNC) state. Changes in the membrane fatty acid (FA) composition of V. vulnificus may be a contributing factor to the ability of this organism to enter into and survive in the VBNC state. This hypothesis was tested by incubating the organism at 5$^{\circ}C$ in arti-ficial sea water and analyzing the cells' FAs during the initial hours of temperature and nutrient down-shift. Prior to downshift, the predominant FAs were 16:0, 16:1 and 18:0. During the first four hours of downshift, statistically significant changes occurred in 15:0, 16:1, 16:0, 17:0, and 18:0. These results indicate that changes in FA composition occur prior to entry of V. vulnificus into the VBNC state, suggesting that the ability to maintain membrane fluidity may be a factor in this physiological response. Cells in which fatty acid synthesis was inhibited did not survive, indicating that active fatty acid metab-olism is essential for entry of cells into the VBNC state.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.6
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• pp.912-917
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• 2021

We investigated the distribution, molecular characteristics, and antimicrobial resistance of Vibrio vulnificus isolated from seawater at Gadeok Island, the Republic of Korea between June to October. Interestingly no isolates were detected between December to February. The detection rate of V. vulnificus was high (80-100%) from July to September 2019 and from June to September 2020. This coincided with the relatively low salinity of the seawater, which ranged from 7.8-29.9 practical salinity units for that period. Additionally, V. vulnificus had a high detection rate at sampling stations near the Nakdong river. The detection rates of virulence genes, such as vvhA, viuB, and vcgC, among the isolates were 97.1%, 44.1%, and 57.4% in 2019 and 100%, 43.0%, and 50.0% in 2020, respectively. Notably, viuB and vcgC were detected in V. vulnificus isolated between June to October when water temperature was above 20℃. The antimicrobial susceptibility analysis of 80 isolates revealed that most of the strains were susceptible to most antimicrobial agents. However, some isolates showed intermediate resistance to cefepime (18.8%), cefoxitin (58.8%), and erythromycin (22.5%). Of note, 3.8% of the tested strains were resistant to cefoxitin. The minimum inhibitory concentration of highly cefoxitin-resistant strains was determined to be less than 32 ㎍/mL.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.430-434
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• 2009

Bacterial 16S rRNA gene sequences have been widely used for the studies on molecular phylogeny, evolutional history, and molecular detections. Bacterial genomes have multiple rRNA operons, of which gene sequences sometimes are variable. In the present study, heterogeneity of the Vibrio 16S rRNA gene sequences were investigated. Vibrio 16S rRNA sequences were obtained from GenBank databases, considering the completion of gene annotation of Vibrio genome sequences. These included V. cholerae, V. harveyi, V. parahaemolyticus, V. splendidus, and V. vulnificus. Chromosome 1 of the studied Vibrio had 7~10 copies of the 16S rRNA gene, and their intragenomic variations were less than 0.9% dissimilarity (more than 99.1% DNA similarity). Chromosome 2 had none or single 16S rRNA gene. Intragenomic 16S rRNA genotypes were detected at least 5 types (V. vulnificus #CMCP6) to 8 types (V. parahaemolyticus #RIMD 2210633, V. harveyi #ATCC BAA-1116). These suggest that Vibrio has high heterogeneity of the 16S rRNA gene sequences.

• The Korean Journal of Food And Nutrition
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• v.10 no.3
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• pp.320-324
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• 1997

The number of Vivrio vulnificus strain 29307 was assessed for oysters treated with acetic, lactic, citric, and alginic acids during storage at 15$^{\circ}C$. When oysters were dipped with 0.5% acetic, 0.5% lactic, or 0.5% citric acids for 3 min, V. vulnificus was not detected after 4 days of storage. V. vulnificus in the treatment of 3% alginic acid (AL) containing 2% acetic acid (AA) was not detected after 2 days of storage, while it was isolated in the controls for 4 days of storage. Based on these results, the combination of AL and AA was more effective for preventing the growth of V. vulnificus in oysters than the treatments of the acid alone.

• Journal of Microbiology and Biotechnology
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• v.7 no.6
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• pp.423-428
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• 1997

Vibrio vulnificus is a halophilic gram-negative human pathogen, which affects people with underlying liver diseases or a suppressed immune system, often leading to primary septicemia with a mortality rate of higher than 60%. In an effort to develop an oral vaccine against V. vulnificus infection, we prepared a whole cell killed vaccine of V. vulnificus on a large scale and compared the immunogenicity and protective efficacy of the vaccine administered in three formulation forms in rabbits. Since V. vulnificus O-antigen serotypes 1, 2, 3, 4, 5, and 7 account for more than 95% of clinical isolates, we prepared cell lysates from these six serotype strains and mixed in equal amounts for a vaccine. The vaccine was administered to rabbits intramuscularly (i.m.), orally as granules or as enteric-coated granules. In rabbits, all three formulation forms elicited a high level of serum IgG antibody reactive not only to the six strains but also to other O-antigen serotypes 6, 8 and 9, indicating cross-reactivities among the strains. Immunotherapeutic efficacy of the antisera was also evaluated by a passive immunization assay, which revealed that the orally immunized antisera as well as the i.m. immunized antisera was protective against a subsequent lethal challenge of V. vulnificus. These data demonstrate that oral immunization with a V. vulnificus whole cell lysate vaccine induced a systemic immune response and suggest the feasibility of development of this vaccine preparation as an oral vaccine.

• Journal of Food Hygiene and Safety
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• v.32 no.2
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• pp.163-169
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• 2017

The purpose of this study was to estimate the antimicrobial activity of Maesil (Japanese apricot, Prunus mume) extract against Vibrio vulnificus. The strains tested in the study were 28 V. vulnificus isolates originated from fish, seawater, mud flat and seawater in fish restaurant. The vvhA gene was detected using real-time PCR and biochemical identification expressed above good identification in 28 isolates of V. vulnificus. All of V. vulnificus used in this study was susceptible to tetracycline and chloramphenicol antibiotics. These two antibiotics were considered to be useful for the treatment of patients. Maesil extracts 2.5% and 5% showed antimicrobial activity against V. cholerae NCCP 13589 and V. parahemolyticus NCCP 11143. V. vulnificus isolate and V. vulnificus NCCP 11135 showed growth inhibition at 1.25%, 2.5% and 5% of Maesil extract, respectively. Compared with V. cholerae and V. parahemolyticus, the antibacterial activity of Maesil extract against V. vulnificus was high. The minimum bactericidal concentration of Maesil extract for V. vulnificus was 1.6%. These results revealed that Maesil extract was found to be very useful for inhibiting the growth of V. vulnificus and can be expected to prevent food poisoning caused by V. vulnificus.

• Journal of Food Hygiene and Safety
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• v.33 no.5
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• pp.412-415
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• 2018

The objective of this study was to establish a quantitative count method of Vibrio vulnificus cells. Plate count method is often used to count the number of V. vulnificus cells using thiosulfate citrate bile salts sucrose (TCBS) agar plate. However, this method is unsuitable for counting V. vulnificus cells due to growth inhibition and cell injuries in TCBS medium. In this study, we suggested a most probable number-polymerase chain reaction (MPN-PCR) method using alkaline peptone water medium for the quantification of V. vulnificus. This MPN-PCR method showed 2 log higher cell number than TCBS agar plate method. Similar results were also found in the control using, Luria-Bertani agar containing 2% NaCl. Thus, this MPN-PCR method can be used a sensitive method for quantitative count of viable V. vulnificus cells in fish and shellfish samples.

• Journal of Life Science
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• v.8 no.2
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• pp.145-157
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• 1998

We isolated 100 Vobrio sp. from marine products and sea from July to September, 1997. We attemped on purification of hemolysin produced by Vibrio sp. The growth, hemolysin production patterns by the 100 strains of Vibrio sp. showed identical, in general. V. unlnificus ys-1 produced hemolysis as the higtest titer. The optimal culture conditions for the hemolysin production by the V. vunificus ys-1 are followings; 1. Hemolysin production was optimal dering the late exponetial phage. 2. Maximal growth, hemolysin production were in heart infusion broth. 3. Maximal yields of hemolysin was obtained when the heart infusion broth had an intial pH of 8.0, 3$0^{\circ}C$, 3% NaCL. Hemolysin was purified from culture filtrate of the strain by ammonium sulfate recipitation, ion exchange and hydrophobic interaction chromatography. The results were as follows; 1. Hemogeneity of the purified hemolysin was demonstrated by revealing single band on SDS-PAGE. The molecular weight of purified hemolysin was 45KDa. 2. The absorbance rattern in ultraviolet wsa typical of those seen with most proteinb with 280nm. 3. Purified hemolysin was atable at 5$0^{\circ}C$ but 7$0^{\circ}C$ of the acivity was lost by heating for 30 min at 6$0^{\circ}C$/ Optimal temperature of purified hemolysin was 35$^{\circ}C$. 4. Purified hemolysin was stable at the pH range of 6~9, but in the less the pH5.0. above the pH 9.0, the hemolysin activity was lost completely.

• Proceedings of the Microbiological Society of Korea Conference
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• 2006.05a
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• pp.29-31
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• 2006

In a previous report, we showed that enzyme $IIA^{Glc}(EIIA^{Glc}$ of Escherichia coli phosphotransferase system (PTS) interacts with and regulates activity of FrsA (fermentation/respiration switch protein). A BLAST search revealed that orthologs of FrsA exist only in some Gram-negative bacteria such as E. coli, Salmonella typhimurium, Shigella flexneri, Yersinia pestis, Vibrio cholerae, Vibrio vulnificus, Vibrio parahemeolyticus, and Photorhabdus luminescens and all of these species are facultative anaerobes belonging to the ${\gamma}-proteobacterial$ group, and most of them are highly pathogenic. Ligand-fishing experiments using $EIIA^{Glc}$ of Vibrio vulnificus ($vEIIA^{Glc}$) as bait revealed that $vEIIA^{Glc}$ also interacts with vFrsA in a phosphorylation state-dependent manner. The frsA mutant of Vibrio vulnificus showed remarkably reduced cytotoxicity to HeLa cells and reduced lethality to mice compared to wild type. Comparison of extracellular proteomes between the mutant and wild type indicated that hemolysin was not produced in the frsA mutant. Characterization of another protein interacting with $vEIIA^{Glc}$ will be discussed.