• Korean Journal of Fisheries and Aquatic Sciences
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• v.27 no.3
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• pp.292-298
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• 1994

An environmental study was done to examine the distribution of Vibrio mimicus in aquatic environments of Kwangan and Minrak beach, Pusan, Korea. Moreover, both bacteriological characteristics and lethal effects of isolated V. mimicus were observed. Sea water samples were collected monthly from January to September, 1993, and quantitatively analyzed for V. mimicus. This organism was isolated from April(water temperature was $16.3^{\circ}C$), whereas it was not isolated when the water temperature fell below $15^{\circ}C$. V. mimicus counts were not remarkably high, however this study at least describes the distribution and occurrence of the possible highest density in aquatic environments of this region. Among the confirmed V. mimicus strains, the author chose the strongest antibiotic resistant bacterium and named it V. mimicus K-1. This strain has antibiotic resistance to colistin, erythromycin, tetracycline, and penicillin, and most isolates had a higher level of antibiotic resistance than V. mimicus ATCC 33653. The optimum growth for V. mimicus K-1 was observed at $37^{\circ}C$, pH 7.5, and $1\%$ NaCl, respectively. This organism was mostly inactivated by Ultra Violet irradiation (30W, $50^{\circ}C$) for 70 seconds and death lethality increased in proportion to treatment temperature ($D_{50}=5.7min,\;D_{60}=\;2.1min,\;and\;D_{70}=0.7min$).

• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.2
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• pp.277-281
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• 1997

The cell density changes of Vibrio mimicus K-1 in sea water and arkshell feeding it were examined at various temperature. The strain was suspended in sterilized sea water and storaged at experimental temperature $(5,\;10,\;15,\;20,\;and\;28^{\circ}C)$). At intervals of up to 10 days, aliquots of each suspension were plated onto BHI agar. At 5 and $10^{\circ}C$, the plate counts of V. mimicus K-1 showed a rapid decline, which 3s known to be a reault of this bacterium's entering into the viable but non culturable state. At 20 and $28^{\circ}C$, however, V. mimicus K-1 are stable over the 10 days experimental periods. V. mimicus K-1 was fed to arkshell, which was subsequently stored at temperatures ranging from 5 to $20^{\circ}C$ for 10 days. The samples of arkshell were homogenized and plated at intervals to determine the cell density of V. mimicus K-1 and total aerobic population of bacteria present. At 5 and $10^{\circ}C$, the numbers of V. mimicus K-1 in sea water rapid decreased over the 10 days experimental periods. However, little change of V. mimicus K-1 density was observed in shellstock arkshell at 5 and $10^{\circ}C$. While, V. mimicus K-1 density was decreased more rapidly to level below limit of dectection in shucked arkshell at same temperature. Incubation at the higher temperature $(20^{\circ}C)$ resulted in large increase in total aerobic bacterial number of shellstock arkshell. These results suggest that even with proper storage, indigenous levels of V. mimicus may remain sufficiently high in shellstock arkshell to produce infection in compromise hosts.

• 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.10 no.5
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• pp.612-619
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• 2000

To investigate the biochemical properties of V. mimicus metalloprotease, whose gene was isolated previously from Vibrio mimicus ATCC33653, overexpression and purification were attempted. The 1.9 kb of open reading frame was amplified by PCR from pVMC193 plasmid which ligated the VMC gene with pUC19 and introduced into Escherichia coli BL21 (DE3) using the overexpression vector, pET22b (+). The overexpressed metalloprotease (VMC) was purified with Ni-NTA column chromatography and characterized with various protease inhibitors, pHs, temperatures, and substrates. The purified VMC showed the proteolytic activity against gelatin, soluble and insoluble collagens, and synthetic peptides. Unlike the observations made with all metalloproteases originated from other Vibrio sp., the VMC did not hydrolyze the casein. The proteolytic activity was critically decreased when the VMC was treated with metal chelating reagents, such as EDTA, 2,2-bipyridine, and 1, 10-phenanthroline. In particular, the 71 kDa VMC exhibited the hemagglutinating activity against human erythrocyte. As the purified VMC was treated with $CuCl_2$ and $NiCl_2$ for the chemical modification of metal binding, the proteolytic activity and hemagglutinating activity were profoundly influenced. The multialignment analysis made on the reported Vibrio metalloproteases showed the difference of amino acid sequence similarity between the two distinctive classes of Vibrio metalloproteases.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.6
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• pp.585-590
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• 2000

A hemolysin producing strain was isolated from Kum rivet estuary located in west part of Korea. In the process of identification the isolated strain was similar to V. mimicus but did not show characteristics of known Vibrio species; therefore, the strain was designated as Vibrio sp. D9 ( V. kumkang) tentatively and further identification study was carried out by comparing its bacteriological characteristics, Morphologically Vibrio sp. D9 was a typical straight roe with a polar flagellium. Among known Vibrio species no identical strains were found when using automatic bacteria identification system ($MicioLog^(TM)$system, release 4.0, Biolog Inc., USA) which evaluated the ability of metabolizing 95 kinds of carbon and nitrogen sources. Vibrio sp. D9 showed 18 and 13 different responses as compared to V. mimicus and V. cholerae, respectively. Clear hemolysis zones were observed with the strain against human and sheep blood agar plate, Hemolytic toxicity was confirmed by strong vascular permeability and fatal toxicity against mouse was also observed. Thus the strain was a pathogenic vibrio. Growth conditions for Vibrio sp. D9 were salinity of $0{\~}5.0{\%}$, pH of $6.4{\~}9.8$, temperature of $15{\~}41^{\circ}C$, respectively.

• Journal of Food Hygiene and Safety
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• v.30 no.3
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• pp.281-288
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• 2015

Vibrio is a genus of Gram-negative, curved, halophilic, and non-spore-forming bacteria. Some of the Vibrio species, such as V. cholerae and V. parahaemolyticus, often contaminate seafood products and occasionally cause human diseases when the seafood products are ingested. A total of 24 Vibrio strains were isolated from shrimp samples on Thiosulphate citrate bile salt sucrose (TCBS) media in this study. All of the 24 isolates were confirmed to belong to the genus Vibrio by using 16S rRNA gene sequence analyses. Vitek 2 system and species-specific polymerase chain reaction (PCR) methods were used to further identify a total of 29 Vibrio strains at the species level, including the 24 shrimp Vibrio isolates and five Vibrio reference strains. The specificities of the two methods to identify Vibrio strains at the species level were compared in this study. The species-specific PCR method was designed to detect five different Vibrio species, such as Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio alginolyticus, and Vibrio mimicus. From the 24 Vibrio shrimp isolates, the Vitek 2 system method could identify 15 (62.5%) strains as Vibrio species and 7 (29.2%) strains as non-Vibrio species, but could not identify the rest 2 (8.3%) strains. But species-specific PCR method could identify 16 (66.7%) strains as Vibrio species and could not identify the rest 8 (33.3%) strains. Among the 24 Vibrio shrimp strains, these two methods could unanimously identify 7 (7/24, 29.2%) strains (2 V. parahaemolyticus, 4 V. alginolyticus, and 1 V. mimicus). Considering that such different identification results were obtained using the two different methods in this study, identification method for Vibrio species must be carefully chosen.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.6
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• pp.545-550
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• 2002

A hemolysin producing bacterial strain which belong to Vibrio species was isolated from the Kum River estuary. In the process of identification, the strain did not show characteristics of known Vibrio species; thus, the strain was designated as Vibrio sp, E10 (V. kunsan) tentatively and further identification study was carried out by comparing its bacteriological characteristics. Morphologically Vibrio sp, E10 was comma shaped rod with a polar flagellium. Clear hemolysis zones were observed with the strain against human and sheep blood agar. Hemollytic toxicity was confirmed by strong vascular Permeability and fatal toxicity against mouse was also observed. Therefore the strain was a pathogenic vibrio. Growth conditions for Vibrio sp. E10 were ranged salinity of 0$\~$$4.5\%$, pH of 6.2$\~$9.2, temperature of 14$\~$42$^{\circ}C$, respectively, 16S rDNA partial sequence of Vibrio sp, E10 showed $99\%$ homology with dozens of V. cholerae species including V, cholerae El Tor N16961 and V, snmisnfus ATCC 33653T. This strain belonged to Proteobacteria; gamma subdivision; Vibrionacea: Vibrio. But, among knorn Vibrio species no identical styains were found when using automatic bacteria identification system ($MicroLog^{TM}$system, release 4.0, Biolog Inc., USA) which evaluated the ability of metabolizing 95 kinds of carbon and nitrogen sources. Vibrio sp, E10 showed 18 and 11 different responses as compared to V. mimicus and V, cholerae, respectively.

• The Journal of the Korean Society for Microbiology
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• v.34 no.6
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• pp.513-518
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• 1999

A total of 113 Vibrio sp. strains were examined for plasmid content which were subjected to digestion with restriction enzymes. About the 55% Vibrio spp. have the plasmid more than one. Most of these plasmid various derivatives ranged from $2.4\;kb{\sim}23\;kb$, especially two strains of V. mimicus and one strain of V. furnissii carried one high-molecular weight plasmid (molecular weight ranging between $70\;kb{\sim}100\;kb$). Results of restriction analysis for plasmid of this three strains were by no means the rule. For detection of tdh and ctx gene, the virulence factor involved in the pathogenesis, we carried out the TDH and CT assay, PCR amplification, and hybridization. A total 11 strains were produced TDH, involved in 9 strains of V. parahaemolyticus and 1 strain of V. alginolyticus from clinical isolates and 1 strains of V. mimicus from environmental isolates. In the experiments of tdh gene detection, in all, 3 strains of V. parahaemolyticus from clinical isolates and 2 strains from environmental isolates could be successfully amplified in 400 bp by PCR. The PCR results were consistent with DNA hybridization tests. In the experiments of CT assay, in all, 3 strains of V. cholerae from clinical isolate and 1 strain of V. cholerae from environmental isolates were observed CT-producing. These CT-producing strains amplified in 302 bp by PCR for the detection of ctx gene. All CT-producing strains hybridized with digoxigenin-labeled DNA probe, while CT-negative strains did not hybridize. Also hybridization tests results for detection of ctx gene consistent with PCR.

• 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.