• Journal of Microbiology and Biotechnology
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• v.23 no.4
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• pp.555-559
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• 2013

Vibrio cholerae O1 and O139 are the major serotypes associated with illness, and some V. cholera non-O1 and non-O139 isolates produce cholera toxin. The present study describes a quantitative polymerase chain reaction (qPCR) assay for the species-specific detection and quantitation of V. cholera using a primer pair based on an outer membrane lipoprotein lolB gene for the amplification of a 195 bp DNA fragment. The qPCR primer set for the accurate diagnosis of V. cholera was developed from publically available genome sequences. This quantitative PCR-based method will potentially simplify and facilitate the diagnosis of this pathogen and guide disease management.

• Journal of Food Hygiene and Safety
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• v.16 no.3
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• pp.241-246
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• 2001

Distribution of pathogenic vibrios in the seawater of live fish tank and effect of environmental factors on their existence were investigated by collecting samples from fish markets and restaurants in 6 different cities. Pathogenic vibrios and coliforms were determined by using the most probable number (MPN) procedure, and aerobic plate count was enumerated by the standard pour plate method. No Vibrio chulerae O1 was detected in all the samples tested. Detection rates of V. cholerae non-O1, V. parahaemolyticus and V. vulnificus in all the smaples tested were 7.7%, 69.2% and 23.1%, respectively. Water temperature and trubidity of the seawater measured were higher in the pathogenic vibrios positive samples than in those negative samples. However, higher salinity and pH were shown in the pathogenic vibrios negative samples than in positive samples. The aerobic plate counts and MPN or total and focal coliforms in the seawater were higher in the presence of pathogenic vibrios than in the absence of pathogenic vibrios. In this study, the presence of pathogenic vibrios in the seawater tested was closely related with other physiochemical parameters and populations of coliforms, indicators far food safety.

• Journal of Microbiology and Biotechnology
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• v.24 no.6
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• pp.725-731
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• 2014

The classical biotype strains of the Vibrio cholerae O1 serogroup harbor the biotype-specific cholera-toxin encoding phage (CTX) $CTX^{cla}$, and the El Tor biotype strains contain CTX-1. Although the classical biotype strains have become extinct, a remnant of classical CTX phage is transferred to the El Tor biotype strains. The prototype El Tor strains, which produce the biotype-specific cholera toxin, are now being replaced by atypical El Tor variant strains producing classical biotype cholera toxin. The genome sequences of the CTX phages in atypical El Tor strains indicate that the CTX phages in atypical El Tor strains are a mosaic of $CTX^{cla}$ and CTX-1. Before the emergence of atypical El Tor stains in the early 1990s, unusual pre-seventh pandemic strains were isolated in the US Gulf Coast between 1973 and 1986. These strains have characteristics of atypical El Tor strains since they are El Tor biotype strains containing $CTX^{cla}$, yet the genome sequence of this CTX phage indicates that it is different from $CTX^{cla}$ and is therefore classified separately as $CTX^{US\;Gulf}$.

• Journal of Microbiology and Biotechnology
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• v.26 no.3
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• pp.627-636
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• 2016

The causative agent of pandemic cholera, Vibrio cholerae, infects the anaerobic environment of the human intestine. Production of cholera toxin (CT), a major virulence factor of V. cholerae, is highly induced during anaerobic respiration with trimethylamine N-oxide (TMAO) as an alternative electron acceptor. However, the molecular mechanism of TMAO-stimulated CT production is not fully understood. Herein, we reveal that CT production during anaerobic TMAO respiration is affected by glucose fermentation. When the seventh pandemic V. cholerae O1 strain N16961 was grown with TMAO and additional glucose, CT production was markedly reduced. Furthermore, an N16961 Δcrp mutant, devoid of cyclic AMP receptor protein (CRP), was defective in CT production during growth by anaerobic TMAO respiration, further suggesting a role of glucose metabolism in regulating TMAO-mediated CT production. TMAO reductase activity was noticeably decreased when grown together with glucose or by mutation of the crp gene. A CRP binding region was identified in the promoter region of the torD gene, which encodes a structural subunit of the TMAO reductase. Gel shift assays further confirmed the binding of purified CRP to the torD promoter sequence. Together, our results suggest that the bacterial ability to respire using TMAO is controlled by CRP, whose activity is dependent on glucose availability. Our results reveal a novel mechanism for the regulation of major virulence factor production by V. cholerae under anaerobic growth conditions.

• Fisheries and Aquatic Sciences
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• v.6 no.3
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• pp.105-109
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• 2003

Fifty four strains of pathogenic vibrios were isolated from the Gwangan Beach from May to October, 2002. The isolated vibrios were composed of 7 different species: Vibrio parahaemolyticus, V. cholerae non-O1, V. alginolyticus, V. vulnificus, V. hollisae, V. fluvialis, ane V. mimicus. In the detection rate, V. parahaemolyticus was most predominant as $46\%$(25/54). From the isolated strains, only 25 strains have hemolytic activity or 25 strains only proteolytic activity on agar plates. Eleven strains showed both hemolytic and proteolytic activity. No strains showed urease activity. All strains of V parahaemolyticus did not show hemolytic activity, while V. cholerae non-O1 strains showed $\beta$ hemolytic activity. Kanagawa phenomena of pathogenic vibrios did not accord with hemolytic activity of the culture supernatant at the late log phase. Some strains showed high hemolytic activity despite having proteolytic activity, but some weak hemolytic activities despite having no proteolytic activity.

• Journal of Environmental Health Sciences
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• v.44 no.2
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• pp.133-142
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• 2018

Objectives: The pathogenic Vibrios genus denotes halophilic bacteria that are distributed in aquatic environments, including both sea and freshwater. Vibrio cholerae, Vibrio vulnificus, and Vibrio parahaemolyticus are the most important species since they can be potent human pathogens and leading causes of septicemia, wound infections, and seafood borne gastroenteritis. The recent emergence of a potential pandemic clone, V. cholera serotype O1 and the cholera outbreak in South Korea in 2016 indicates the importance of consistent surveillance of pathogenic Vibrio genus within coastal areas. Methods: The present study was undertaken to determine where and how vibrios live in the aquatic environment adjacent to coastal areas of South Korea. For this survey, a total of 838 samples were obtained at 35 different sites in South Korean coastal areas during the period from January 2016 to December 2016. Pathogenic vibrios was determined using the real-time PCR method, and its clones were isolated using three selective plating media. We also monitored changes in seawater and atmospheric temperature, salinity, turbidity, and hydrogen ion concentration at the collection points. Results: The total isolation rates of V. vulnificus, V. cholera (non-pathogenic, non-O1, non-O139 serogroups), and V. parahaemolyticus from seawater specimens in 2016 were 14.2, 13.48, and 67.06%, respectively. Conclusions: The isolation rates of pathogenic vibrios genus showed a positive correlation with temperature of seawater and atmosphere but were negatively correlated with salinity and turbidity.

• Proceedings of the Korean Environmental Sciences Society Conference
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• 2001.05a
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• pp.240-241
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• 2001

자연계로부터 Vibrio sp.에 항균활성을 갖는 균을 검색하여 분리하고 분리균의 동정결과 Pseudomonas aeruginoso로 동정되었으며, 분리한 균의 항균성 물질의 최적 생산조건을 검토하며, 항균성 물질을 정제하여 그 안정성과 항균작용기작에 대해 조사한 결과 pH와 열에 대한 광범위한 안정성을 보였으며, 항균물질이 V. cholerae non-O1 ATCC 25872 세포에 대해 뚜렷한 용균작용을 하는 것을 관찰하였다.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.5
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• pp.557-566
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• 1996

An ecological study on pathogenic vibrios was done in the aquatic environments of southern coast of Korea during summer in 1995, to investigate the distribution and relationship between pathogenic vibrio and zooplankton. Furthermore, special emphasis was given to study on the efforts of zooplankton existence on the wintering of Vibrio cholerae in the aquatic region in Korea. During the study period, pathogenic vibrios were isolated from the samples such as V. parahaemolyticus, V. vulnificus, V. mimicus, and V. cholerae non O1, but V. cholerae O1 was not detected in any sample submitted in this study. Adsorption ratio of V. parahaemolyticus onto zooplankton was higher than that of E. coli. The efficiency of adsorption was found to be on the concentration of NaCl and other ions found in sea water. For example, adsorption ratio of V. parahaemolyticus were $75\%\;at\;5\%_{\circ}$ of NaCl solution and $55\%$ at same salinity of diluted sea water, but those were decreased as $20\%\;and\;7\%\;at\;15\%_{\circ}$ salinity of NaCl solution and diluted sea water, respectively. In addition, survival period of pathogenic vibrio was extended in the presence of live copepods at $25^{\circ}C$, but zooplankton existence has no significant effect on the survival rate at $5^{\circ}C$ in closed microcosm and also microalgae and dead copepods do not affect on the survival of V. parahaemolyticus. According to these experimental results, zooplankton has positive effects on the growth and survival rate of pathogenic vibrios in sea water during the summer season, but copepods have no significant effects on the growth and survival rate of them in winter season in Korea. Finally, authors suggest that V. cholerae is not able to over winter with zooplankton in adjacent sea water in Korea.

• Proceedings of the Microbiological Society of Korea Conference
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• 2000.05a
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• pp.58-65
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• 2000

Diarrheal diseases are a major cause of both illness and death in developing countries and are caused by rotavirus, Shigella spp., Salmonella spp., enterotoxigenic Escherichia coli (ETEC), and Vibrio spp. In this study, for the development of vaccine against diarrheal diseases caused by Shigella sonei, Salmonella typhimurium, E. coli O157, and Vibrio cholerae, cloning and nucleotide sequence analysis of genes and characteristics of their gene products in E. coli were performed. For construction of attenuated strain of S. sonnei KNIH104 and Salmonella typhimurium KNIH100, the aroA genes were cloned, respectively. The recombinant plasmid $_pJP{\Delta}A45$ containing aroA deleted region and suicide vector $(_pJP5603)$ was constructed. The aroA gene deleted mutants were constructed using this recombinant plasmid. For cloning gene encoding antigenic region of E. coli O157 KNIH317, the O-antigen synthesis gene cluster and sit gene was cloned. The E. coli XL1-Blue cells harboring this recombinant plasmid showed cytotoxicity in Vero cells. The ctx gene was cloned for tile purpose of antigenic region against V. cholerae KNIH002. Sequence analysis confirmed that the virulence gene cassette was consisted of ace, zot, ctxA and ctxB genes.

• Fisheries and Aquatic Sciences
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• v.7 no.1
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• pp.10-15
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• 2004

A total of 52 pathogenic Vibrio strains was isolated from the Gwangan Beach during summer in 2003. The isolated vibrios were composed of 6 different species: V. parahaemolyticus, V. cholerae non O1, V. fluvialis, V. vulnificus, V. alginolyticus, and V. mimicus. V. parahaemolyticus was most predominant as $46\%$ (24/52), V. cholerae non O1 was the second with $23\%$ (12/52), and V. fluvialis was the third with $17\%$ (9/52). Among the isolated strains, 22 strains showed hemolytic, proteolytic or ureolytic activity. Eight strains showed both hemolysin and protease activities, and either 6 strains showed only hemolysin activities and 7 strains only protease activities. Only one strain of V. parahaemolyticus isolates showed urease activity. The urease-positive V. parahaemolyticus strain (V. parahaemolyticus S25) showed the same biochemical characteristics as the reference strain, V. parahaemolyticus KCTC 2471 (urease­negative) except for urease production. To compare the degree of virulence of Vibrio strains having different pathogenic factors, hemolysin, protease, or urease-positive strains were injected into groups of 10 each of ICR mice (7- to l0-week-old male). The lethal rate of urease-positive V. parahaemolyticus S25 was significantly high, being $70\%$. Protease-positive strains showed $40-60\%$ of lethal rate. Hemolysin-positive strains showed no mortality, similar to non-pathogenic V. parahaemolyticus KCTC 2471 and V. parahaemolyticus FM12.