• 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 7^th to the 20^th 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.

• 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)^-α] 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.

• 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.

• 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.

• 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.

• 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.

• 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.

• 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.

• Korean Journal of Microbiology
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• v.35 no.3
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• pp.205-210
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• 1999

16brio cholerne is an important pathogenic organism that causes dimhea in human beings. V ciaoleroe KNIH002 was isolated from patients suffering with dian.heal disease in Korea. From Southern hybridization using the amplified PCR product of 307 bp as a probe. which was obtained from PCR reaction using primer detecting cholera toxin gene, we have found that the c b gene located in 4.5-kb fragmenl double digested with Pstl and BgllI of the chromosome. Therefore, we made mini-libraries of the isolate using PstI and Bgm restriction endonuclease and pBluescript SKU(+) vector. As a result. we cloned 4.5-kb PstI-BglII fragment containing the c a gene encoding a cholera toxin from the constructed mini-libraries of V olzolerae KNlH002 by colony hybridization using the same probes. This recombinant plasmid was named pCTX75. E. coii XL1- Blue harboring pCTX75 showed the cytotoxicity on Chinese Hamster Ovary cells. From the sequencing of he cloned recombinant plasmid, we confinned that it has virulence gene cassette consisting of ace, zot, ctx.4 and cf"~B gene. The ace and zot genes were composed of 291 hp and 1.200 bp with ATG initiation codon and TGA lennination codon, respectively. Nucleotide sequence of the ace gene exhibited 100% identity with that of V cholera E7946 El Tor Ogawa strains. But, nucleolide and amino acid sequence comparison of the zot gene exhibited 99% and 98.8% identity with that of V cholerae 395 Classical Ogawa stram, respectively. Specially. the Ala-100, Ala-272 and Ala-281 sites of Zoi polypeptide presented in V choleme 395 Classical Ogawa strain are replaced by Val in V cholerae KNIH002.