• 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.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.2
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• pp.222-227
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• 2001

The members of the genus Vibrio include harmless aquatic strain as well as strains capable of causing infections in human and fish. Pathogenic mechanisms are only understood for Vibrio cholerae O1 and O139 and not for the majority of Vibrio species. Twelve clinical and nonclinical strains were examined by in vitro and in vivo experiments for the importance of extracellular enzymes as a virulence determinant of Vibrio species. In vivo cytotoxicity assay was performed by injecting approximately $10^{8}$ cells/mL into mice (BALB/c). V. harvyi and V. vulnificus showed 100% lethality within 3hr after bacterial injection. V. fluvialis and four strains of V. parahaemolyticus showed 50% lethality within 4hr. V. mimicus, V. alginolyticus and V. furnissii revealed 30% lethality within 9hr. Nonclinical strains, V. campbellii and V. ordalii, did not show any lethality. In vitro protease and hemolytic activities were also good indicators for clinical and nonclinical strains of Vibrio species. The clinical strains showed much higher activities than nonclinical strains. The activity of some clinical strains of re-isolates was evidently increased. Most clinical strains had $\beta$ hemolytic activity. The results demonstrate that the prevalent distribution of extracellular proteases in pathogenic Vibrio sp. implies their importance as a virulence determinant.

• Journal of Microbiology and Biotechnology
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• v.22 no.8
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• pp.1107-1112
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• 2012

Outbreaks of foodborne diseases associated with Vibrio species such as V. parahaemolyticus, V. vulnificus, and V. cholerae frequently occur in countries having a dietary habit of raw seafood consumption. For rapid identification of different Vibrio species involved in foodborne diseases, whole-cell protein pattern analysis for 13 type strains of 12 Vibrio species was performed using SDS-PAGE analysis. Pathogenic Vibrio species such as V. parahaemolyticus, V. vulnificus, V. cholerae, V. alginolyticus, V. fluvialis, and V. mimicus were included in the 12 Vibrio species used in this study. Each of the 12 Vibrio species showed clearly specific band patterns of its own. Two different strains of V. parahaemolyticus showed two different SDS-PAGE whole-cell protein patterns, giving the possibility of categorizing isolated strains in the same V. parahaemolyticus species into two subgroups. The 36 Vibrio isolates collected from sushi restaurants in Busan were all identified as V. parahaemolyticus by comparing their protein patterns with those of Vibrio type strains. The identified isolates were categorized into two different subgroups of V. parahaemolyticus. The whole-cell protein pattern analysis by SDS-PAGE can be used as a specific, rapid, and simple identification method for Vibrio spp. involved in foodborne diseases at the subspecies level.

• 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 Life Science
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• v.27 no.10
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• pp.1161-1167
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• 2017

In order to find a new antimicrobial bacterium, we performed screening for antimicrobial activity of bacteria isolated from the eggs of a sea hare. The newly identified strain was designated as Phaeobacter inhibens KJ-2, based on the biochemical characterization and 16S rRNA gene sequence analysis. A colony of P. inhibens KJ-2 showed a circular and ruler-like smooth form at the edge, and a brown color. However, when maintained with a longer incubation time, its coloring was transformed into dark brown. From the result of SEM, P. inhibens KJ-2 is a bacillus which has a length of $0.8{\sim}1.0{\mu}m$ and a width of $0.4{\sim}0.6{\mu}m$. The optimal growth and antimicrobial activity were observed by shaking the culture for 24 hr at $20^{\circ}C$, which showed potent activity against pathogenic bacteria including Vibrio logei, Vibrio campbellii, Vibrio mimicus, Vibrio vulnificus, and Vibrio salmonicida. The antimicrobial activity was proportional to the amount of produced acylated homoserine lactones (AHLs). Therefore, we suggest that production of antimicrobial materials from P. inhibens KJ-2 is regulated by Quorum sensing (QS).

• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.4
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• pp.335-343
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• 2009

Antimicrobial resistance patterns of Vibrio species isolated from brackish water in Geoje, Tongyeong and Goseong, Gyeongsangnamdo province into which streams, sewage and leachate all flowed. Only 19 strains (10.7%) of 177 V. parahaemolyticus were susceptible to 15 antimicrobials. 146 strains (69.5%) proved resistant against more than one antimicrobial and 12 strains (6.8%) were multi-drug resistant. The resistance rate of 152 strains were 85.9% against AM and 26.6% against RA, 16.4% against AN, 13.6% against Sand 13.0% against TMP. 86 strains of 129 V. cholerae non-O1 (66.7%) were susceptible to antimicrobials and 31 strains (24.0%) were resistant to more than one antimicrobial and 12 strains (9.3%) were multi-drug resistant. The antimicrobial resistance rate of 129 strains against 15 antimicrobials, with the exception of C, CIP, E and GM, i.e. 11 antimicrobials, was 0.7-16.2%, 16.2% of 129 strains proved resistant against RA and 13.9% against AM, 9.3% against TMP, 7.7% against SXT and 6.9% against TE. 19 of 49 strains of V. mimicus (38.8%) were susceptible to antimicrobials and 31 strains (61.2%) were resistant against more than one antimicrobial; none of the strains were multi-drug resistant. 15 strains of V. mimicus were resistant against only RA, AmC and TE. The resistance rate was 59.2% against RA (highest) 4.1% against AmC and 2.0% against TE.

• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.355-362
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• 2016

The phosphomannomutase/phosphoglucomutase gene (pmm/pgm) of Vibrio anguillarum (the causative agent of fish vibriosis) was cloned, and the open reading frame corresponded to a protein with 446 amino acids. The pmm/pgm gene showed a significant degree of sequence homology with the previously reported genes from V. mimicus, V. vulnificus, V. splendidus, and V. harveyi, with 92.3%, 91.4%, 89.9%, and 89.9% amino acid identity, respectively. By reverse transcriptase-polymerase chain reaction, we found that the pmm/pgm gene was upregulated under cold stress condition. The PMM/PGM protein is known to catalyze the interconversion between mannose-1-phosphate and mannose-6-phosphate or glucose-1-phosphate and glucose-6-phosphate, which are important intermediates for lipopolysaccharide (LPS) biosynthesis. To confirm the role of PMM/PGM in the LPS biosynthetic pathway, we constructed a knock out mutant by homologous recombination. The respective LPSs were isolated from the V. anguillarum wild-type and mutant strains, and changes were compared by subjecting them to sodium dodecyl sulfate polyacrylamide gel electrophoresis. Based on the different patterns of the LPSs, we expect the pmm/pgm gene to have an important role in LPS biosynthesis. The pmm/pgm-deficient mutant of V. anguillarum will contribute to further studies about the role of LPS in V. anguillarum pathogenesis.

• Journal of Food Hygiene and Safety
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• v.25 no.4
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• pp.367-375
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• 2010

The bacteriological and physiochemical analysis of sea water and sediments in Tongyeong harbor was conducted to evaluate sanitary conditions. The samples were collected at 8 stations established once a month from June, 2008 to May, 2009. During the study period, the range of temperature was from 6.7 to $25.2^{\circ}C$, transparency ranged from 1.2 to 2.6 m, chemical oxygen demand ranged from 1.90 to 2.92 mg/L, dissolved oxygen ranged from 6.2 to 10.5 mg/L, dissolved nitrogen ranged from 0.052 to 0.098 mg/L, phosphate ranged from 0.044 to 0.065 mg/L, respectively. Seafood, if eaten raw, carries the risk of food poisoning. Seafood poisoning is often cause by pathogenic microorganism originating from fecal contamination, such as Salmonella sp., Shigella sp. and norovirus. Fecal coliforms are an important indicator of fecal contamination. Therefore, data on fecal coliform are very important for evaluating the safety of fisheries in coastal areas. So, we investigated the sanitary indicate bacteria. The coliform group and fecal coliform MPN's of sea water in Tongyeong harbor were ranged from < 1.8~22,000/100 mL (GM 164.9 MPN/100 mL) and < 1.8~7,900 MPN/100 mL (GM 33.7 MPN/100 mL), respectively. Total coliform were detected 97.0% in 96 of samples and 68.9% of total coliforms were fecal coliforms. These results similar to another seawater detection ratio of total coloforms and fecal coliforms. The Vibrios was isolated and identified with VITEK system. Four hundred eighty strains that were obtained from sea water samples in Tongyeong harbor Detection ratio Vibrio alginolyticus, 34.2%, Vibrio parahaemolyticus, 13.8%, Vibrio vulnificus 10.0%, and V. mimicus 12.5% respectively. Vibrio cholerae O1, was not detected. During the study period, the ranges of water content, ignition loss, COD, and acid volatile sulfates in sediments in Tongyeoung harbor were 41.0~57.4%, 7.8~10.5%, 6.51~9.30 mg/g, 0.04~0.09 mg/g, respectively. Heavy metals in sediment of Tongyeoung harbor were Cd, $0.10{\pm}0.05$; Cu, $4.79{\pm}8.20$; As, $1.95{\pm}0.17$; Hg, $0.10{\pm}0.07$; $Cr^{6+}$, $0.34{\pm}0.12$; Zn, $125.33{\pm}16.40$; Ni, $16.43{\pm}1.93$ mg/kg.

• Journal of Microbiology and Biotechnology
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• v.25 no.1
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• pp.119-126
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• 2015

Among the various human growth factors, epidermal growth factor (hEGF, consisting of 53 amino acids) has various effects on cell regeneration, stimulation of proliferation, migration of keratinocytes, formation of granulation tissues, and stimulation of fibroblast motility, which are important for wound healing. Owing to their multiple activities, EGFs are used as pharmaceutical and cosmetic agents. However, their low productivity, limited target specificity, and short half-life inhibit their application as therapeutic agents. To overcome these obstacles, we fused the collagen-binding domain (CBD) of Vibrio mimicus metalloprotease to EGF protein. About 18 or 12 amino acids (aa) (of the 33 total amino acids), which were essential for collagen-binding activity, were combined with the N- and C-termini of EGF. We constructed, expressed, and purified EGF (53 aa)-CBD (18 aa), EGF (53 aa)-CBD (12 aa), CBD (18 aa)-EGF (53 aa), and CBD (12 aa)-EGF (53 aa). These purified recombinant proteins increased the numbers of cells in treated specimens compared with non-treated specimens and control hEGF samples. The collagen-binding activities were also evaluated. Furthermore, CBD-hybridized hEGF induced phosphorylation of the EGF receptor. These results suggested that these fusion proteins could be applicable as small therapeutic agents in wound tissue healing.