• Microbiology and Biotechnology Letters
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• v.31 no.4
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• pp.334-341
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• 2003

This study was carried out to investigate the cause of cholera outbreak in Gyeongbuk province in 2001.90 strains of Vibrio cholerae O1 El Tor serotype Inaba were isolated from diarrheal patients. By multiplex-PCR, all of the isolated strains revealed positive for detection ctxA, hlyA and tcpA genes. There were DNA sequence difference of the cholera-toxin subunit A gene and subunit B gene between isolated V. cholerae O1 and the strain of GenBank. In analysis of PFGE patterns, all of the isolated strains were showed the same DNA fragments. We also collected plankton samples in the east coast of Gyeongbuk to isolate V. cholerae O1 and V. cholerae O139 from August to October 2002. The samples were examined to detect the rfb gene and cholera-toxin gene by multiplex-PCR. The cholera-toxin gene was detected and then we tried to isolate V. cholerae O1 and V. cholerae O139, but they were not isolated.

• Journal of Food Hygiene and Safety
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• v.13 no.2
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• pp.149-154
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• 1998

The authors attempted utilization of fatty acid composition of vibrios as a tool for identification of the strains. Fatty acid of 49 strains of Vibrio cholerae non-O1, V. cholerae O1, V mimicus, V vulinificus and V parahaemolyticus was analyzed by gas-liquid chromatography column. According to the statistical analysis of the fatty acid data, the relationship between the Vibrio species and serotypes of the strains was discussed. Forty one kinds of fatty acid were detected from the tested strains and 35 kinds of fatty acids among the detected fatty acids were significant factors to identify the vibrios. The predominant fatty acids were 16:0, 16:1 cis 9, 18:1 trans 9/6/cis 11 and 15:0 iso 2OH/16:1 cis 9 as above about 20% in total. Fatty acid compositions of the Vibrio species were an important factor in identifying their subspecies either predominant fatty acids or minor ones. According to the analysed results by a conventional statistical processing method (UPGMA) and prepared dendrogram, V cholerae non-01 had more closer relationship with V. mimicus compared with V. cholerae 01. Moreover, the distribution of hydroxy acid was a significant factor for identifying V cholerae subspecies. Comprising all the 10 serotypes detected from V. cholerae non-01 examined such as O2, O5, O8, O10, O14, O27, O37, O39, O45 and O69, we could group them into seven subspecies by cluster analysis with the similarity value of fatty acid composition as above 92%. It means that there is a significant relationship between serotypes and fatty acid composition of V. cholerae. These results indicated that numerical analysis of fatty acid composition data of V cholerae non-01 could classifY them into subspecies, and also which may provide a useful epidemiologic information or a basis for further analysis such as PCR and DNA probe analysis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.1
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• pp.15-22
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• 1995

To study ecological properties of Vibrio cholerae non-Ol and Vibrio mimicus which have been described as new food poisoning bacteria recently, the influence of factors such as temperature, salinity, pH and chemical oxygen demand (COD) on detection rate and density of these bacteria were evaluated. Fifty four seawater samples and 49 bottom deposit samples from estuary of Kum river from March 26th, 1993 to February 22nd, 1994 were used for this study. The detection rate of V cholerae non-O1 were $16.7\%$ for seawater and $10.2\%$ for bottom deposit, respertively. The total detertion rate of V. cholerae non-O1 $(11.7\%)$ was a little higher than V mimicus $(10.7\%)$. Both V choierae non-O1 and V. mimicus were mainly detected in estuary water of which showed temperature $24^{\circ}C$ above and salinity $10\%o$ below. These bacteria were also detected in bottom deposit on January when the water temperature was $3.5^{\circ}C$. From these results, we supposed that temperature, salinity and organic material were important factors to growth of V. cholerae non-O1 and V. mimicus. V cholerae non-O1 might be grown better than V. mimicus under the fluctuating aquatic environmental condition such as salinity.

• Microbiology and Biotechnology Letters
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• v.31 no.1
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• pp.13-17
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• 2003

The studies were carried out to identification of biochemical characteristics and serogroup of 55 Vibrio cholerae non-01 isolated from sea waters and shellfish from Apr., 1996 to Nov., 1996 in Korea coastal, and clinical sources from 1984 to 1996 in Korea hospitals. The results were as followed: V. cholerae non-O1 isolated 21 (10%) of 206 samples and the distribution in Kusan area was highest (16%) compared with those of other area, and its were not isolated during low water temperature of 13-18$^{\circ}C$. Of 55 strains, a strain was delayed sucrose utilization for 48hr in the biochemical characteristics. Fifty five V. cholerae non-O1 were differentiated 15 types by serogroup test and O14 of sea waters were 76% of highest followed by 14 clinical sources were O8.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.4
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• pp.550-555
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• 1997

Vibrio cholerae non-O1 (V. cholerae non-O1) was previously called nonagglutinable or noncholera vibrios, since it fails to react with polyvalent O1 antisera. This organism is biochemically and genetically indistinguishable from V. cholerae O1 except serological difference. V. cholerae non-O1 strains are often detected in the environment including bays, estuaries, and fresh water, and also found in food. Therefore it is designated food borne bacterium in Japan. However, research papers on V. cholerae non-O1 are very rare in Korea. In order to investigate bacteriological characteristics of V. cholerae non-O1, we isolated V. cholerae non-O1 from the environmental sea water. Among the isolated V. cholerae non-O1 strains, we selected the strain which had the most strong hemolytic activity, named as V. cholerae non-O1 FM-3. The optimum growth conditions of V. cholerae non-O1 FM-3 were $37^{\circ}C$ and pH 8.5 in BHI broth (containing $0.5\%$ sodium chloride), and it grew better than V. cholerae non-O1 ATCC 25872. But both were not able to grow in BHI broth added $5.0\%$ of sodium chloride or adjusted to pH 5.0. According to the experimental results on the susceptibility test against various antibiotics, there were no significant differences between the isolated strain and reference strain (V. cholerae non-O1 ATCC 25872). Most of the antibiotics examined had bacteriostatic action against V. cholerae non-O1 FM-3 while vancomycin, oxacillin, colistin, polymyxin B, and sulfadiazine had no bacteriostatic activity.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2001.05a
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• pp.217-219
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• 2001

비브리오균은 현재까지 약 30여종이 알려져 있는 그람음성의 종속영양세균으로, 이 중 12종이 사람에게 질병을 일으킨다고 보고되고 있으며, 우리 나라에서 문제가 종종 발생하는 비브리오균은 패혈증 비브리오, 장염 비브리오 그리고 V.cholerae non-O1을 들 수가 있다. V. cholerae O1과 형태학적, 생화학적, 생물학적 성상은 일치하지만, 콜레라균의 항혈청에는 응집하지 않는 일군의 세균을 V. cholerae non-O1이라고 부른다. (중략)

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.1
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• pp.60-66
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• 1995

Vibrio cholerae non-O1 and Vibrio mimicus, food poisoning bacteria, have detected frequently in fresh water and brackish water. To estabilsh prevention measures of food poisoning outbreak by these bacteria, the adaptability and population changes were examined in fresh water, brackish water $(10\%o\;NaCl)$ and seawater $(30\%o\;NaCl)$. Both species poorly survived as temperature increased regardless of water types employed. However, survival time was the shortest in fresh water and longest in seawater at $4^{\circ}C$. In case of brackish water, the bacteria survived best at $15^{\circ}C$ and population were varied only in small numbers. Any significant difference was not observed to both species with respect to water types and temperatures except V. mimicus survived about 5-6 days longer in brackish water. In conclusion, V. cholerae non-O1 and V. mimicus were not likely to be recovered In normal fresh water, brackish water and seawater, and both biological and physicochemical factors could affect survival of these species.

• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.914-920
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• 2011

Vibrio cholerae utilizes mannitol through an operon of the phosphoenolpyruvate-dependent phosphotransferase (PTS) type. A gene, mtlD, encoding mannitol-1-phosphate dehydrogenase was identified within the 3.9 kb mannitol operon of V. cholerae. The mtlD gene was cloned from V. cholerae O395, and the recombinant enzyme was functionally expressed in E. coli as a $6{\times}$His-tagged protein and purified to homogeneity. The recombinant protein is a monomer with a molecular mass of 42.35 kDa. The purified recombinant MtlD reduced fructose 6-phosphate (F6P) using NADH as a cofactor with a $K_m$ of $1.54{\pm}0.1$ mM and $V_{max}$ of $320.8{\pm}7.81\;{\mu}mol$/min/mg protein. The pH and temperature optima for F6P reduction were determined to be 7.5 and $37^{\circ}C$, respectively. Using quantitative real-time PCR analysis, mtlD was found to be constitutively expressed in V. cholerae, but the expression was up-regulated when grown in the presence of mannitol. The MtlD expression levels were not significantly different between V. cholerae O1 and non-O1 strains.

• Journal of Life Science
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• v.12 no.6
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• pp.669-675
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• 2002

Introduction of sliced raw fish meat(SRFM) to fast food business has been considered seriously. However bacteria causing food poisoning should be controlled. Organic acids such as vinegar and lactic acid used in the sauce for SRFM were evaluated for their antibacterial activities. At low concentration levels of vinegar and lactic acid exerted strong antibacterial activities toward Vibriu sp.. In contrast, in case of Salmonella typhimurium and Escherichia coli O157:H7 low anitbacterial activities were observed even at relatively high concentrations. Minimum inhibitory concentrations(MIC) of vinegar for V. vulnificus, V. cholerae non-O1, V. parahaemolyticus, V. mimicus, S. typhimurium and E. coli O157:H7 were 16, 18, 16, 12, 26, and $20{\mu}\ell /m\ell, respertively. MIC of lactic acid for V. vilnificus, V. cholerae non-O1, V. parahaemolyticus, V. mimicus, S. typhimurium and E. coli O157:H7 were 20, 25, 25, 25, 40, and $35{\mu}\ell /m\ell, respectively. In case of vinegar bactericidal concentration upon 10 second contact for V. vulnificus, V. cholerae non-O1, V. parahaenolyticus, V. mimicus and E. coli O157:H7 were 8, 14, 10, 4, and 48%, respectively; however, even at 50% colony of S. typhimurium was observed. In case of lactic acid any colony was observed for V. vulnificus, V. cholerae non-O1, V. parahaemolyticus, V. mimicus, S. typhimurium and E. coli O157:H7 at the concentration of 2, 3, 4, 3, 14, and 17%, respectively. Vinegar and lactic acid of low concentration inhibited the growth of Vibrio sp., food poisoning pathogen in SRFM; in contrast, at high concentration these organic acids inhibited Salmonella sp. and Escherichia sp., food poisoning pathogen in other than SRFM.

• Journal of Food Hygiene and Safety
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• v.15 no.2
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• pp.128-136
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• 2000

Twenty-four Vibrio strains were isolated from imported frozen seafoods and identified according to their physiological and biochemical properties. They included two V cholerae non-01 sp., two V. diazotrophicus sp., one V. hollisae sp., five V. natriegens sp., eight V. fluvialis sp., and four V. nereis sp.. Two of them were not identified as Vibrio species. When these strains were tested using API-2OE kit fur identification, however, only the results for two V. cholerae and five of the V. fluvialis strains matched the results obtained previously. Due to the importance of detecting V cholerae from foods, phylogenetic identification of the strains was attempted based on restriction fragment length polymorphism (RFLP) of the 16S rDNAs amplified by PCR. The results suggested that the two strains had identical RFLP patterns which were more closely related to that of V. proteolyticus than V. cholerae. The problems associated with identification of pathogens originated from seafoods demand development of accurate and rapid identification methods.