• Journal of Environmental Science International
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• v.1 no.2
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• pp.93.2-97
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• 1992

The naturally occurring plasmids of Vibrio species have been isolated in part to investigate their genetic traits. Among six different Vibrio species tested, Vibrio anguillarum, Vibrio fluvialis, Vibrio vulnificus, Vibrio mimicus and Vibrio furnissi did not show any presence of plasmid. One environmental isolate of Vibrio pnrahemolyticus harboring plasmid was observed. The isolated plasmid was 8.7 kb by analysis with restriction endonuclease digestion. No common feature was shown relationships between the presence of plasmid and resistance against commonly used antibiotic compounds from the tested Vibrios. Key words . plasmid, Vibrio anguillarum, Vibrio fluvialis, Vibrio uulnincus, Vibrio mimicus, Vibrio furnissi, Vibrio parahemolyticus.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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• v.1 no.2
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• pp.93-97
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• 1997

The naturally occurring plasmids of Vibrio species have been isolated in part to investigate their genetic traits. Among six different Vibrio species tested, Vibrio anguillarum, Vibrio fluvialis, Vibrio vulnficus, Vibrio mimicus and Vibrio furnissi did not show any presence of plasmid. One environmental isolate of Vibrio parahaemolyticus harboring plasmid was observed. The isolated plasmid was 8.7 kb by analysis with restriction endonuclease digestion. No common feature was shown relationships between the presence of plasmid and resistance against commonly used antibiotic compounds from the tested Vibrios.

• Journal of Life Science
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• v.6 no.4
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• pp.241-249
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• 1996

A collagenase was isolated from the culture filtrate of Vibrio mimicus (ATCC 33658). The enzyme was purified to homogeneity by ammonium sulfate precipitation and DEAE-Sephadex A-50 chromatography, which an activity recovery of 22%. The molecular weight of the purified enzyme was estimated to be 42 kDa by SDS-polyacrylamide gel electrophoresis and gel filtration, indication a monomer structure. The optimum pH and temperature od the enzyme for insoluble collagen (Type I) were around 7.75 and 28$\circ$C, respectively. Some chelating agents and serine protease inhibitor inactivated the enzyme, but L-cysteine and histidine did not affect the activity. The amino acid composition indicated that the collagenase contained high amounts of amino acid residues of glycine and alanine. The K$_{m}$ and R$_{cat}$/K$_{m}$ values for the collagenase, using insoluble collagen (type I) as substrate, were 2.86 mg/ml and 972.28 U/mg-protein, respectively.

• 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 ($D_{50}=5.7min,\;D_{60}=\;2.1min,\;and\;D_{70}=0.7min$).

• Journal of Food Hygiene and Safety
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• v.12 no.1
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• pp.9-14
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• 1997

Vibrio mimicus is a closely related species with V. cholerae, and has been reported to be associated with gastrointestinal infections. Although extraintestinal infections of these vibrios have also been reported in Japan and Southeast Asia. But little research papers on V. mimicus was reported in Korea. Therefore, we tried to isolate V. mimicus from the environmental sea water from April to July in Pusan, Korea. Among the isolated strains, we selected the strongest hemolytic strain and then named V. mimicus SM-9. In this paper, we checked the antibiotic susceptibility and psychrotrophic characteristics of the isolated strain. Hemolytic activity of the hemolysin produced by the isolated strain was also measured. V. mimicus was not detected from the sea water samples in April and May, but its detection rate was relatively high in June and July in Pusan, Korea. The bacteriological characteristics of V. mimicus SM-9 were Gram-negative rods, motile, oxidase positive, Voges-Proskauer negative and sucrose negative. In 23 kinds of antibiotics susceptibility test, V. mimicus SM-9 showed susceptibility to the most of antibiotics submitted while it was resistive against lincomycin, oxacillin, rifampin and vancomycin. Hemolytic activity of the hemolysin produced by V. mimicus SM-9 was highest in stationary phase of the growth curve in BHI broth at 37$^{\circ}C$ and its activity was reached 18 HU per $m\ell$ of culture supernatant. For checking the psychrotrophic property of V. mimicus SM-9, the decreasing rate of the strain in phosphate buffer solution and yellowtail flesh homogenate was examined during the storage at 4, 0, -4 and -2$0^{\circ}C$. The decreasing rates of the selected strain stored in phosphate buffer solution were greater than those in fish homogenate. Decreasing rates of V. mimicus SM-9 stored in phosphate buffer solution were not significantly different by the storage temperatures. The viable cell counts of the strain were decreased as 5 log cycles after 120 hours at all the tested temperatures. While decreasing numbers of the strain in fish homogenates were 2*4 log cycles after 120 hours. The decreasing pattern of the strain numbers were very slow after 200 hours at all the stored temperatures.

• 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.

• The Journal of the Korean Society for Microbiology
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• v.34 no.5
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• pp.461-470
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• 1999

Growth under conditions of iron-restriction and the production of siderophore was examined in Vibrio mimicus ATCC 33653. This strain grew and multiplied in the presence of the high-affinity iron chelators ethylenediamine-di (o-hydroxyphenylacetic acid). Chrome azurol S (CAS) agar and solution were used to detect the production of siderophore under these condition. Siderophore could be detected in the iron-restricted culture supernatants. The siderophore was extracted from iron-restricted culture supernatants by phenol-chloroform-ether method and purified by Dowex ion-exchange and Sephadex G-25 gel filtracton chromatography. The purified siderophore was confirmed by paper chromatography and HPLC. The Purified siderophore enhanced the growth of V. mimicus when the bacterium was grown in iron limited medium. Injection of both the siderohore and the bacteria to mice resulted in more rapid death than that of the only bacteria. However, the siderophore did not show lethality to mice and any toxicity to cell line like HeLa and U937.

• 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 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.