• Title/Summary/Keyword: Vibrio Cholerae

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Intracellular Expression of CTB in Vibrio cholerae Strains in Laboratory Culture Conditions

  • Hunseok Choi;Seonghyeon Son;Donghyun Lee;Jonghyun Bae;Eunyoung Seo;Dong Wook Kim;Eun Jin Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.736-744
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    • 2023
  • The introduction of the toxT-139F allele triggers the expression of TCP (toxin co-regulated pilus) and CT (cholera toxin) under simple laboratory culture conditions in most Vibrio cholerae strains. Such V. cholerae strains, especially strains that have been used in OCVs (oral cholera vaccines), can induce antibody responses against TCP in animal models. However, CT produced in these V. cholerae strains is secreted into the culture medium. In this study, V. cholerae strains that can express intracellular CTB under the control of the toxT-139F allele have been constructed for potential application in OCVs. First, we constructed a recombinant plasmid directly linking the ctxAB promoter to ctxB without ctxA and confirmed CTB expression from the plasmid in V. cholerae containing the toxT-139F allele. We constructed another recombinant plasmid to express NtrCTB, from which 14 internal amino acids-from the 7th to the 20th amino acid-of the leader peptide of CTB have been omitted, and we found that NtrCTB remained in the cells. Based on those results, we constructed V. cholerae strains in which chromosomal ctxAB is replaced by ntrctxB or ntrctxB-dimer. Both NtrCTB and NtrCTB-dimer remained in the bacterial cells, and 60% of the NtrCTB-dimer in the bacterial cells was maintained in a soluble form. To develop improved OCVs, these strains could be tested to see whether they induce immune responses against CTB in animal models.

Variations in the Antivirulence Effects of Fatty Acids and Virstatin against Vibrio cholerae Strains

  • Donghyun Lee;Jayun Joo;Hunseok Choi;Seonghyeon Son;Jonghyun Bae;Dong Wook Kim;Eun Jin Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.9
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    • pp.1757-1768
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    • 2024
  • The expression of two major virulence factors of Vibrio cholerae, cholera toxin (CT) and toxin co-regulated pilus (TCP), is induced by environmental stimuli through a cascade of interactions among regulatory proteins known as the ToxR regulon when the bacteria reach the human small intestine. ToxT is produced via the ToxR regulon and acts as the direct transcriptional activator of CT (ctxAB), TCP (tcp gene cluster), and other virulence genes. Unsaturated fatty acids (UFAs) and several small-molecule inhibitors of ToxT have been developed as antivirulence agents against V. cholerae. This study reports the inhibitory effects of fatty acids and virstatin (a small-molecule inhibitor of ToxT) on the transcriptional activation functions of ToxT in isogenic derivatives of V. cholerae strains containing various toxT alleles. The fatty acids and virstatin had discrete effects depending on the ToxT allele (different by 2 amino acids), V. cholerae strain, and culture conditions, indicating that V. cholerae strains could overcome the effects of UFAs and small-molecule inhibitors by acquiring point mutations in toxT. Our results suggest that small-molecule inhibitors should be examined thoroughly against various V. cholerae strains and toxT alleles during development.

Quantitative Microbial Risk Assessment of Pathogenic Vibrio through Sea Squirt Consumption in Korea (우렁쉥이에 대한 병원성 비브리오균 정량적 미생물 위해평가)

  • Ha, Jimyeong;Lee, Jeeyeon;Oh, Hyemin;Shin, Il-Shik;Kim, Young-Mog;Park, Kwon-Sam;Yoon, Yohan
    • Journal of Food Hygiene and Safety
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    • v.35 no.1
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    • pp.51-59
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    • 2020
  • This study evalutated the risk of foodborne illness from Vibrio spp. (Vibrio vulnificus and Vibrio cholerae) through sea squirt consumption. The prevalence of V. vulnificus and V. cholerae in sea squirt was evaluated, and the predictive models to describe the kinetic behavior of the Vibrio in sea squirt were developed. Distribution temperatures and times were collected, and they were fitted to probabilistic distributions to determine the appropriate distributions. The raw data from the Korea National Health and Nutrition Examination Survey 2016 were used to estimate the consumption rates and amount of sea squirt. In the hazard characterization, the Beta-Poisson model for V. vulnificus and V. cholerae infection was used. With the collected data, a simulation model was prepared and it was run with @RISK to estimate probabilities of foodborne illness by pathogenic Vibrio spp. through sea squirt consumption. Among 101 sea squirt samples, there were no V. vulnificus positive samples, but V. cholerae was detected in one sample. The developed predictive models described the fates of Vibrio spp. in sea squirt during distribution and storage, appropriately shown as 0.815-0.907 of R2 and 0.28 of RMSE. The consumption rate of sea squirt was 0.26%, and the daily consumption amount was 68.84 g per person. The Beta-Poisson model [P=1-(1+Dose/β)] was selected as a dose-response model. With these data, a simulation model was developed, and the risks of V. vulnificus and V. cholerae foodborne illness from sea squirt consumption were 2.66×10-15, and 1.02×10-12, respectively. These results suggest that the risk of pathogenic Vibrio spp. in sea squirt could be considered low in Korea.

Pulsed-Field Gel Electrophoresis-Based Molecular Typing Reveals a Shift in the Major Type of Vibrio cholerae O1 Isolated in Korea

  • Kim, Seong-Han;Kim, Jun-Young;Kang, Yeon-Ho;Park, Yong-Keun;Lee, Bok-Kwon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1814-1818
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    • 2006
  • Vibrio cholerae O1 El Tor isolates (n=242), collected in Korea between 1991 and 2002, were classified by NotI-digested pulsed-field gel electrophoresis (PFGE). The major types were Al before 1998 and B after 1999, among both domestic and imported cases. The prevalent PFGE types among domestic cases were consistent with the prevalent types among imported cases in the same year. These results suggest a close relationship between the domestic and imported cases of cholera in Korea.

Identification of the Vibrios Isolated from a Shellfish, Sunset Shell, (Soletellina olivacea) (빛조개(Soletellina olivacea)로부터 분리된 비브리오의 생화학적 성상)

  • 이훈구
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.185-191
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    • 1999
  • This study was conducted to investigate the vibrio flora in an edible shellfish. sunset shelfish. Soletelliim olivacen. which were collected in the estuarine area. Dadaepo near Nakdong River in Korea lkoin January 1997 to November 1997. Including five pathogemc vibrios (Vibrio alginolyticus, Vibrio pamhaemol~~licz~s, Vibrio cholerae non-01. Vibrio vulnificus, and Vihrio jl~~vinlis), a lotal of eight species of vlbr~os (Vi61-io splendidrrs biovar I, Vibrio splendidus biovar 11, Vibrio snlrnonicida and Vibrio tr,~biasllii) were identified from the sunset shellfish by heir biochemical characters. The isolation of Vihrio pamhaemolyricns, which is known not to grow below $15^{\circ}C$, in winter season indicates that the sunset shelllish is one oT the natural owl.- wintering hosts for Vibrio parahuemolyticus.

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Susceptibility of Vibrio vulnificus to Human Serum Bactericidal Activity and Effect of Vibrio Infections on Hematocrit Value in Mice (Vibrio vulnificus의 인혈청살균력에 대한 감수성과 Vibrio 감염이 마우스의 Hematocrit치에 미치는 영향)

  • Ha, Tai-You;Im, Suhn-Young;Chun, Sang-Nam;Kim, Chul-Kee
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.3
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    • pp.355-361
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    • 1986
  • Vibrio vulnificus, a halophilic Vibrio has gained worldwide attention as a cause of severe and frequently fatal wound infections and life-threatening septicemia. For this reason V. vulnificus is thought to produce extreme hemoconcentration and rapid death after infection, and because V. vulnificus is thought to be less susceptible to bactericidal activity of normal human serum, the present study was undertaken to assess the susceptibility of V. vulnificus to human serum bactericidal activity with that of V. parahemolyticus and V. cholerae and to assess the effect of Vibrio species, Salmonella typhimurium and E. coli on hematocrit values in experimentally infected mice. Serum bactericidal activity against both V. vulnificus and V. cholerae was higher than against V. parahemoltyicus. Survival of the test strains in heat-inactivated human serum was greater than that in heat-uninactiveted serum. Both V. parahemolyticus and V. cholerae produced slight hemoconcentration within 2 to 6 hr after intraperitoneal injection of $10^7$ viable bacteria into mice. In contrast, V. vulnificus, S. typhimurium and E. coli produced hemodilution rather than hemoconcentration after 4 or 6 hr after infection. With these results the author can conclude that V. vulnificus is more susceptible to serum bactericidal activity than other Vibrio species, and V. vulnificus did not produce hemoconcentration.

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Effects of flaC Mutation on Stringent Response-Mediated Bacterial Growth, Toxin Production, and Motility in Vibrio cholerae

  • Kim, Hwa Young;Yu, Sang-Mi;Jeong, Sang Chul;Yoon, Sang Sun;Oh, Young Taek
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.816-820
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    • 2018
  • The stringent response (SR), which is activated by accumulation of (p)ppGpp under conditions of growth-inhibiting stresses, plays an important role on growth and virulence in Vibrio cholerae. Herein, we carried out a genome-wide screen using transposon random mutagenesis to identify genes controlled by SR in a (p)ppGpp-overproducing mutant strain. One of the identified SR target genes was flaC encoding flagellin. Genetic studies using flaC and SR mutants demonstrated that FlaC was involved in bacterial growth, toxin production, and normal flagellum function under conditions of high (p)ppGpp levels, suggesting FlaC plays an important role in SR-induced pathogenicity in V. cholerae.

pVC, a Small Cryptic Plasmid from the Environmental Isolate of Vibrio cholerae MP-1

  • Zhang, Ruifu;Wang, Yanling;Leung, Pak Chow;Gu, Ji-Dong
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.193-198
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    • 2007
  • A marine bacterium was isolated from Mai Po Nature Reserve of Hong Kong and identified as Vibrio cholerae MP-1. It contains a small plasmid designated as pVC of 3.8 kb. Four open reading frames (ORFs) are identified on the plasmid, but none of them shows homology to any known protein. Database search indicated that a 440 bp fragment is 96% identical to a fragment found in a small plasmid of another V. cholerae. Further experiments demonstrated that a 2.3 kb EcoRI fragment containing the complete ORF1, partial ORF4 and their intergenic region could self-replicate. Additional analyses revealed that sequence upstream of ORF1 showed the features characteristic of theta type replicons. Protein encoded by ORF1 has two characteristic motifs existed in most replication initiator proteins (Rep): the leucine zipper (LZ) motif located at the N-terminal region and the alpha helix-turn-alpha helix motif (HTH) located at the C-terminal end. The results suggest that pVC replicates via the theta type mechanism and is likely a novel type of theta replicon.

Distribution of Pathogenic Vibrio spp. in Seawater of the Geum River Estuary Area, West Coast of Korea (금강 하구 해역의 해수에 병원성 비브리오균(Vibrio spp.)의 분포)

  • Park, Seon-A;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.55 no.6
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    • pp.844-849
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    • 2022
  • 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.