• 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 the Korea Academia-Industrial cooperation Society
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• v.20 no.5
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• pp.102-108
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• 2019

Melaleuca alternifolia contains terpineol-4, cineol, sesquiterpenes etc., and has a germicidal effect and skin moisturizing effect. It also has the characteristics of relieving acne inflammation, treating dandruff, relieving pain, and relieving depression. In this study, an antimicrobial substance extracted from tea tree using an organic solvent (methanol, dichloromethane, ethyl acetate) and hydrothermal extraction method. And confirmed the antimicrobial activity of each extract. In order to verify the antimicrobial activity, nine pathogenic bacteria (Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa and Vibrio parahaemolyticus) were used. The antimicrobial activity of each extracts were confirmed by the commonly used disc diffusion method. The results showed that the fraction extracts of ethyl acetate and methanol had antimicrobial effects against V. parahaemolyticus and S. aureus. Using these results, we confirmed the antimicrobial activity of each fraction extracts and hot water extracts against V. parahaemolyticus. After the treat of samples, we confirmed at over 99.9 % of antimicrobial activity. In case of antifungal activities, we confirmed of preservation effect during over 45 hours. Based on the results of this research, further studies will be conducted to confirm the possibility of development as a new antimicrobial agent.

• Journal of Food Hygiene and Safety
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• v.7 no.4
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• pp.137-142
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• 1992

The distribution of V. parahaemolyticus was surveyed from various kinds of shells in Kunsan Bay from July to September, 1987. 1be morphological, biochemical and serological characteristics of the isolated strains were studied. The results were as follows: 1. 41 strains were isolated from 1,350 specimens of shells (Crassostrea gigas, Tapes philippinarum, Meretrix Iusoria) 2. The isolation rates of V. parahaemolytieus were 3% in July, 3.8% in August, and 2.2% in September, respectively. 3. V. parahaemolytkus was more frequently isolated from Kunsan (20 strains) than Bideukgi (12 strains) and Gae Hwa-do (9 strains). 4. V. parahaemolyticus was more frequently isolated from C gigas (23 strains) than other shells. 5. Kanagawa hemolysis reactions were all negative. 6. 6 Strains positive to K pooled antiserum included K IV, K V, K VI and K vn type.

• Korean journal of food and cookery science
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• v.13 no.3
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• pp.330-337
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• 1997

The sensitivity of various pathogenic bacteria (Aeromonas hydrophila, Estherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus 196E, Salmonella typhimurium and Vibrio parahaemolyticus) to the spices, allspice, clove, oregano, and thyme, was tested. Tryptic soy broth (TSB) containing 0∼2% (w/v) of spices was inoculated with 10sup 5/∼10$\^$6/ cells/$m\ell$ of each bacterium and incubated at 35$^{\circ}C$ for 24 hr. The growth of pathogenic bacteria was inhibited with increasing concentrations of spices in the culture broth. At 2％ spice concentration, Gram positive bacteria were more sensitive than Gram negative bacteria with the exception of V. parahaemolyticus. Clove had the highest antibacterial activity, followed by allspice and oregano. At the concentration of 0.3%, clove inhibited the growth of all strains tested. Kanagawa-positive strain of V. parahaemolyticus displayed the highest sensitivity to clove and allspice. Thyme was the least effective for growth inhibition, while 1％ clove killed all pathogens tested.

• Journal of Microbiology and Biotechnology
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• v.33 no.9
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• pp.1162-1169
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• 2023

16S rRNA short amplicon sequencing-based microbiota profiling has been thought of and suggested as a feasible method to assess food safety. However, even if a comprehensive microbial information can be obtained by microbiota profiling, it would not be necessarily sufficient for all circumstances. To prove this, the feasibility of the most widely used V3-V4 amplicon sequencing method for food safety assessment was examined here. We designed a pathogen (Vibrio parahaemolyticus) contamination and/or V. parahaemolyticus-specific phage treatment model of raw oysters under improper storage temperature and monitored their microbial structure changes. The samples stored at refrigerator temperature (negative control, NC) and those that were stored at room temperature without any treatment (no treatment, NT) were included as control groups. The profiling results revealed that no statistical difference exists between the NT group and the pathogen spiked- and/or phage treated-groups even when the bacterial composition was compared at the possible lowest-rank taxa, family/genus level. In the beta-diversity analysis, all the samples except the NC group formed one distinct cluster. Notably, the samples with pathogen and/or phage addition did not form each cluster even though the enumerated number of V. parahaemolyticus in those samples were extremely different. These discrepant results indicate that the feasibility of 16S rRNA short amplicon sequencing should not be overgeneralized in microbiological safety assessment of food samples, such as raw oyster.

• Microbiology and Biotechnology Letters
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• v.44 no.3
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• pp.261-267
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• 2016

The present study examined the effect of the dietary administration of a halophilic bacterium Bacillus sp. Mk22 on the growth improvement of black tiger shrimp, Penaeus monodon, and reduced shrimp infection by Vibrio parahaemolyticus and white spot syndrome virus (WSSV). The shrimp were fed 45 days using three experimental diets: no addition (control), commercial probiotic, and Bacillus sp. Mk22. The shrimp treated with the halophilic bacterium Mk22 showed a significant improvement of growth (7.1 ± 0.21 g), survival (94.3 ± 0.58%), weight gain (178 ± 4.93 g), and reduced feed conversion rate (0.8 ± 0.03 g) compared with the shrimp in the other groups. The shrimp treated with Bacillus sp. Mk22 also showed a lower Vibrio count (0.02 ± 0.01 × 10² CFU/ml) in the shrimp culture water compared with the other groups. The shrimp in the three groups were challenged with either Vibrio or WSSV. For the Vibrio infection, no mortality was observed from water infection or oral feeding infection in the commercial probiotic group and Mk 22 group. For the WSSV infection, a 68% survival rate from water infection and 20% survival rate from oral feeding infection was observed on day 45 in the Mk22 group, while 100% mortalities were recorded at a much earlier time in both the control and commercial probiotic groups. The antioxidant enzyme activities, indicators of oxidative stress, such as catalase and superoxide dismutase, significantly decreased in both the Vibrio and WSSV-infected Mk22 groups compared with the other groups, indicating that Bacillus sp. Mk22 was effective in reducing oxidative stress, possibly due to the reduced infection.

• Journal of Microbiology
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• v.35 no.2
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• pp.87-91
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• 1997

Monoclonal antibodies were prepared in order to an assay method for Vibrio vulnificus. Sixteen mouse ybridoma cell lines were established by immunization of whole cell antigen to BALB/c mice, fusion with SP2/O myeloma cells, and cloning. Most of them secreted IgM.lambda. antibodies. A sandwich enzyme-linked immunosorbent assay was developed with rabbit anti-V. vulnificus polyclonal antibodies as capture antibody, an IgM monoclonal antibody as detector antibody, and goat anti-mouse IgM-alkaline phosphatase conjugate as developer antibody. The range of detection was 10$\^$4/ to 10 V. vulnificus cells per microplate well. When four related Vibrio species were tested for cross-reactions, V. parahaemolyticus showed 3.5% reactively and V. carchariae, V. fluvialis, and V. furnisii showed negligibal (<1%) cross-reactivity.

• 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.

• The Journal of the Korean Society for Microbiology
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• v.18 no.1
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• pp.1-9
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• 1983

The author studied on the distribution of V. parahaemolyticus among sea water, sea mud, and various marine products in Jeju, Keoje, Namhae, Yockji, Busan, and Masan on the southern seas of Korea from winter in 1981 to summer in 1982. The author studies for the isolated strains to bacteriological identification Kanagawa phenomenon(hemolytic activity) on Modified Wagatuma blood agar plates and serotypes with anti V. parahaemolyticus. The results obtained were as follows:. 1. V. parahaemolyticus was isolated in 713 strains(28.3%) among 2519 total specimens of sea water, sea mud, and various marine products. 2. The isolation rates of V. parahaemolyticus in summer season were higher than in spring and winter season. Above results were 304 strains(32.6%) among 932 specimens in summer, 160 strains (28.1%) among 570 specimens in spring, 149 strains(14.6%) among 1017 specimens in winter, respectively. 3. The hemolysis on Modified Wagatuma agar added human erythrocytes was 66.0% of positive Kanagawa phenomenon, and was 34.0% of negative Kanagawa phenomenon, respectively. 4. The distributions of serotypes of V. parahaemolyticus isolated were from KI to K VIII of K pooling antisera. The results were 6 strains(6.6%) on K 1 of K I, 14 strains(l5.5%) on K 17 of K II, 26 strains(28.8%) on K 28 of K IV. 10 strains(11.1%) on K 32 of KV, 4 strains(4.4%) on K 39 of KV, 8 strains(8.8%) on K 42 of K VI, 2 strains(2.2%) on K 48 of K VII, 1 strain(1.1%) on K 50 of K II and 7 strains(7.7%) on K 55 of K VII, respectively. 5. V. parahaemolyticus was more frequently isolated from sea mud than sea water and various marine prdoucts in winter season. 6. There was no great difference as far as the distribution of V. parahaemolyticus concerned in Jeju, Keoje, Namhae, Yockji, Busan and Masan of the southern seas of Korea.

• 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.