• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.903-913
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• 2011

Vibrio parahaemolyticus, which causes gastroenteritis, wound infection, and septicemia, has two sets of type III secretion systems (TTSS), TTSS1 and TTSS2. A TTSS1-deficient vcrD1 mutant of V. parahaemolyticus showed an attenuated cytotoxicity against HEp-2 cells, and a significant reduction in mouse lethality, which were both restored by complementation with the intact vcrD1 gene. V. parahaemolyticus also triggered phosphorylation of mitogen-activated protein kinases (MAPKs) including p38 and ERK1/2 in HEp-2 cells. The ability to activate p38 and ERK1/2 was significantly affected in a TTSS1-deficient vcrD1 mutant. Experiments using MAPK inhibitors showed that p38 and ERK1/2 MAPKs are involved in V. parahaemolyticus-induced death of HEp-2 cells. In addition, caspase-3 and caspase-9 were processed into active forms in V. parahaemolyticus-exposed HEp-2 cells, but activation of caspases was not essential for V. parahaemolyticus-induced death of HEp-2 cells, as shown by both annexin V staining and lactate dehydrogenase release assays. We conclude that secreted protein(s) of TTSS1 play an important role in activation of p38 and ERK1/2 in HEp-2 cells that eventually leads to cell death via a caspase-independent mechanism.

• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1423-1431
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• 2012

The inhibitory effect of IgY against Vibrio parahaemolyticus and Vibrio vulnificus responsible for seafood-borne diseases was investigated in this study. Water-soluble fractions (WSF) of protein containing IgYs were isolated from the egg yolk of hens initially immunized with formalin-inactivated V. parahaemolyticus or V. vulnificus. Protein, total and specific IgY contents of the WSF were determined. The inhibitory and protective effects of IgYs on the growth of V. parahaemolyticus and V. vulnificus were assayed in liquid medium and in mice. IgYs showed high affinity to their corresponding antigens with high titer from day 28 onwards. Protein contents and total IgY concentrations remained stable throughout the immunization period, whereas specific IgY concentrations increased steadily and reached a plateau at day 49. Specific IgY powder (150 mg/ml) significantly inhibited further multiplication of both V. parahaemolyticus and V. vulnificus in liquid medium as compared with the control IgY. The bacteria count in mice feces was lower in mice pretreated with specific IgYs than in those pretreated with PBS or control IgY. Higher survival of mice was observed in the experimental groups pretreated with either anti-V. parahaemolyticus (75% survival) or anti-V. vulnificus (87% survival) IgYs, compared with those in the control groups pretreated with PBS or nonspecific IgY. All mice in the control groups died within three days after bacteria inoculation; hence, the protective effect of specific IgYs against infection caused by V. parahaemolyticus and V. vulnificus was demonstrated.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.118-124
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• 2013

Vibrio parahaemolyticus in uncooked seafood causes acute gastroenteritis. The microorganism has two sets of type III secretion systems and two hemolysins. When it injects its effector proteins into a host cell via type III secretion system 1, one of the type III secretion systems induces secretion of interleukin (IL)-8, a proinflammatory chemokine, through the phosphorylation of ERK 1/2 and p38 MAPK. Although probiotics have beneficial effects on hosts and can help control some infectious diseases, there is little research on the efficacy of probiotics in V. parahaemolyticus infection. Here we pretreated V. parahaemolyticus-infected human intestinal epithelial cells with heat-killed Lactobacillus brevis KB290, a probiotic isolated from fermented vegetables (traditional Japanese pickles) and utilized as an ingredient of beverages and supplementary foods, and demonstrated its efficacy in enhancing IL-8 secretion from V. parahaemolyticus-infected cells. Among the three heat-killed lactic acid bacterial strains we tested, L. brevis KB290 induced the highest level of IL-8 secretions in the infected cells. Relative to control cells (Caco-2 cells pretreated with PBS), V. parahaemolyticus-infected Caco-2 cells pretreated with heat-killed L. brevis KB290 secreted IL-8 earlier, although concentrations were similar 450min after infection. Heat-killed L. brevis KB290 pretreatment also induced earlier ERK 1/2 phosphorylation, greater p38 MAPK phosphorylation, and enhanced IL-8 mRNA expression. Heat-killed L. brevis KB290 accelerated IL-8 secretion, a host cell immune response, in V. parahaemolyticus-infected cells. We consider this to be beneficial because IL-8 plays an important defensive role against infection, and would contribute to the repair of injured epithelial cells.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.52 no.2
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• pp.114-120
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• 2019

In the case of being contaminated with pathogenic Vibrio, shellfish pose a serious threat for public health. This study was conducted to investigate the presence of Vibrio parahaemolyticus and V. vulnificus in the seawater, bottom deposit and shellfish samples collected from the Gomso Bay, west coast of Korea. V. parahaemolyticus (30-80 CFU/g) was detected from seawater Jul. to Oct. and highest at Sep. V. parahaemolyticus was detected less than 10 CFU/g from seawater at Nov. with $14.6^{\circ}C$ of seawater temperature. V. parahaemolyticus (180-1,850 CFU/g) was detected more than 100 CFU/g from 3 kinds of shellfish, Short-necked clam Ruditapes philippinarum, Corb shell Cyclina sinensis, and Surf clam Mactra veneriformis, Jul. to Oct. and was highest in Surf clam at Oct. V. parahaemolyticus was detected less than 20 CFU/g from all tested shellfish at Nov. V. vulnificus (1.8-2.7 MPN/100 mL) was detected from seawater Jul. to Oct. and was not detected at Nov. V. vulnificus (18-236.7 MPN/100 g) was also detected from 3 kinds of shellfish Jul. to Oct. and was highest in Corb shell at Jul. V. vulnificus was not detected from all tested shellfish at Nov.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.5
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• pp.456-460
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• 2009

Biofilm is a structured community of microorganism encapsulated within a self-developed polymeric matrix and adherent to a living or a solid surface. In this study, we investigated the effects of various substrates on the formation of biofilm in Vibrio parahaemolyticus. We found that biofilm formation profoundly increased in a substrate, that consisted of calcium chloride, calcium nitrate, and calcium sulfate in 1% peptone water. On the other hand, a dramatic reduction in biofilm formation was observed in a substrate, that consisted of glucose and ferric chloride in LB broth. These results suggest that V. parahaemolyticus prefer to form a biofilm on the surface of a crustacean or a clam, where calcium ion is rich, and also where seawater temperature is relatively lower. In contrast, high levels of glucose in a crustacean or a clam body resulting from increased seawater temperature, can make V. parahaemolyticus detach from it and lead to free floating.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.43 no.6
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• pp.637-641
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• 2010

The antibiotics-resistance profiles of 28 strains of Vibrio parahaemolyticus isolated from seawater were investigated. All of the strains studied were resistant to ampicillin (100%), but susceptible to 12 other antibiotics. The minimum inhibitory concentration (MIC) of V. parahaemolyticus to ampicillin was as high as $1,024-2,048\;{\mu}g{\cdot}mL^{-1}$. The phenotype of strain 8 changed from ampicillin-resistant to susceptible with an in-frame deletion mutant of VPA0477, a putative ${\beta}$-lactamase gene, and the MIC for ampicillin of the mutant strain was $1{\mu}g{\cdot}mL^{-1}$. In conclusion, our findings suggest that the VPA0477 gene acts as a ${\beta}$-lactamase in ampicillin-resistant V. parahaemolyticus strains.

• Journal of Food Hygiene and Safety
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• v.9 no.3
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• pp.141-149
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• 1994

To investigate the distribution of Vibrio parahaemopyticus in the southern 4 coastal areas, Vibrio parahaemolyticus was isolated from seawater, shellfish and sediment from May to October in 1991, and antimicrobial effect of grapefruit seed extract (GFSE) on the growth of isolated strains were examined. In the 120 sample from 4 coastal areas, 16 strains of Vibrio parahaemolyticus were isolated and identified. The distribution serotype of isolated strains was 10 types of monovalent k-antiserum. Especially k-5 and k-28 were highly distribyted with 3 and 4 strains. 31.3% of isolated strains showed positive on Kanagawa phenomenon test. All isolated Vibrio parahaemolyticus were sensitive to chloramphenicol and gentamycin, 5 and 6 strains were resistant to streptomycin and colistin, respectively. Isolated strains were compared with geographical, month and sample. The distribution of 16 isolated Vibrio parahamolyticus was high at Hadong with 50%(8 strains), on July with 43.8%(7 strains) and from seawater with 37.5%(6 strains) respectively. Minimal inhibitory level of GFSE to Vibrio parahaemolyticus was 50 ppm. With 100 ppm treatment of GFSE, the destroy of cell membrane function, outflow of cell ingredients and ghost morphology of cell were investigated.

• Journal of Microbiology and Biotechnology
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• v.21 no.10
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• pp.1026-1035
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• 2011

Vibrio parahaemolyticus, which owes its origin to the marine environment, is considered as one of the most common causes of infectious diarrhea worldwide. The present study investigated the pathogenicity of V. parahaemolyticus against the model organism, Caenorhabditis elegans. Infection in the host was localized with GFP-tagged V. parahaemolyticus using confocal laser scanning microscopy. The times required for causing infection, bacterial load in intestine, chemotactic response, and alteration in pharyngeal pumping were analyzed in the host system. In addition, the regulation of innate immune-related genes, lys-7, clec- 60, and clec-87, was analyzed using real-time PCR. The role of immune-responsible pmk-1 was studied using mutant strains. The pathogenicity of environmental strain CM2 isolated from the Gulf of Mannar, India was compared with that of a reference strain obtained from ATCC. The pathogen infected animals appeared to ward off infection by up-regulating candidate antimicrobial genes for a few hours after the exposure, before succumbing to the pathogen. For the first time, the pathogenicity of V. parahaemolyticus at both the physiological and molecular levels has been studied in detail using the model organism C. elegans.

• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.6
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• pp.1163-1166
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• 2002

Antimicrobial activity of silver ion was tested against 3 kinds of food-borne microorganisms-Salmonella typhimurium, Staphylococcus aureus and Vibrio parahaemolyticus-using paper disk and broth medium methods. In paper disk method, silver ion showed antimicrobial activity against S. typhimurium and V. parahaemolyticus at the concentration above 2 ppm and 10 ppm, respectively, where as it was not detected in S. aureus with 20 ppm of silver ion concentration. In broth medium, the growth of S. typhimurium and V. Porahaemolyticus could be retarded at 0.3 ppm and 0.5 ppm of silver ion concentration respectively. In the presence of 1.0 ppm of silver ion, the growth of S. typhimurium was inhibited completely. In S. aureus, the growth was retarded at 5 ppm and was inhibited at l5 ppm completely.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.43 no.2
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• pp.125-130
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• 2010

A trimethoprim resistant Vibrio parahaemolyticus, which cause acute gastroenteritis in humans, was isolated from farmed fish and seawater. The resistance profiles of isolated V. parahaemolyticus and their correlation with mobile elements were investigated. All of the V. parahaemolyticus were resistance to both rifampin and trimethoprim. The presence of class I integron was confirmed by PCR. PCR-amplified inserted gene cassettes contained aminoglycoside aac6-II, rifampin arr-3 and trimethoprim dfrA27 resistance genes. This study indicated that class I integron mainly contributed to the circulation of trimethoprim resistance determinants in V. parahaemolyticus.