• 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.

• Journal of Microbiology and Biotechnology
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• v.30 no.6
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• pp.856-867
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• 2020

Vibrio parahaemolyticus is a major gastroenteritis-causing pathogen in many Asian countries. Antimicrobial resistance in V. parahaemolyticus has been recognized as a critical threat to food safety. In this study, we determined the prevalence and incidence of antimicrobial resistance in V. parahaemolyticus in the southern Fujian coast, China. A total of 62 isolates were confirmed in retail aquatic products from June to October of 2018. The serotype O3:K6 strains, the virulence genes tdh and trh, antibiotic susceptibility and molecular typing were investigated. Then plasmid profiling analysis and curing experiment were performed for multidrug-resistant strains. The results showed that the total occurrence of V. parahaemolyticus was 31% out of 200 samples. Five strains (8.1%) out of 62 isolates were identified as the V. parahaemolyticus O3:K6 pandemic clone. A large majority of isolates exhibited higher resistance to penicillin (77.4%), oxacillin (71%), ampicillin (66.1%) and vancomycin (59.7%). Seventy-one percent (44/62) of the isolates exhibited multiple antimicrobial resistance. All 62 isolates were grouped into 7 clusters by randomly amplified polymorphic DNA, and most of the isolates (80.6%) were distributed within cluster A. Plasmids were detected in approximately 75% of the isolates, and seven different profiles were observed. Seventy-six percent (25/33) of the isolates carrying the plasmids were eliminated by 0.006% SDS incubated at 42℃, a sublethal condition. The occurrence of multidrug-resistant strains could be an indication of the excessive use of antibiotics in aquaculture farming. The rational use of antimicrobial agents and the surveillance of antibiotic administration may reduce the acquisition of resistance by microorganisms in aquatic ecosystems.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.3
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• pp.220-226
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• 2014

Sixty-seven Vibrio parahaemolyticus isolates from surface seawater from the Wando area, on the southern coast of Korea, were analyzed for their susceptibility to 15 different antimicrobials and the presence of virulence genes. According to the disk diffusion susceptibility test, all of the strains studied were resistant to ampicillin and oxacillin, while decreasing percentages were resistant to vancomycin (64.2%), streptomycin (56.7%), amikacin (31.3%), kanamycin (22.3%), cephalothin (20.9%), erythromycin (10.4%), ciprofloxacin (4.5%), and tetracycline (3.0%). All of the strains were susceptible to five antimicrobials: chloramphenicol, gentamycin, nalidixic acid, sulfamethoxazole/trimethoprim, and trimethoprim. Fifty-nine isolates (88.1%) were resistant to three or more classes of antimicrobial and defined as multidrug resistant, and two strains were resistant to seven antimicrobial agents. The minimum inhibitory concentration (MIC) of the 67 V. parahaemolyticus isolates to ampicillin and oxacillin ranged from 512-2,048 and $64-512{\mu}g/mL$, respectively. All 67 isolates were also examined for the presence of the tdh and trh virulence genes using the polymerase chain reaction (PCR). However, no isolates possessed either tdh or trh. The VPA0477 (${\beta}$-lactamase) gene, present in all of the tested strains, was validated as a new specific marker gene in PCR assays for the accurate detection and identification of V. parahaemolyticus.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.6
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• pp.597-604
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• 2011

Using 108 strains of Vibrio parahaemolyticus isolated from seawater, we investigated ampicillin-resistance profiles and the genetic characterization of ${\beta}$-lactamase (VPA0477). All of the strains studied, except one strain, were resistant to ampicillin. However, the strain that was susceptible to ampicillin had the same ${\beta}$-lactamase gene as the ampicillin-resistant strains. We compared ${\beta}$-lactamase promoter region sequences among five strains, including both ampicillin-resistant and -susceptible strains. In the susceptible strain, a nucleotide at position -19 in the methionine initiation codon for ${\beta}$-lactamase was not present in the ampicillin-resistant strains. The genes in the region containing the gene VPA0477 were present in all of the tested strains, and LA-PCR analysis showed that the distance between VPA0474 and VPA0479 in all of the V. parahaemolyticus samples was precisely 5.7 kb. In V. parahaemolyticus ${\beta}$-lactamase, four important structural features that are conserved in Class A ${\beta}$-lactamases were present in the deduced amino acid sequences. Taken together, our study demonstrates that V. parahaemolyticus ${\beta}$-lactamase is included in the Class A ${\beta}$-lactamase group, and some nucleotides within the promoter region are of particular importance for ${\beta}$-lactamase activity.

• Journal of Life Science
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• v.13 no.3
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• pp.308-313
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• 2003

The collagenase gene from Vibrio parahaemolyticus 04 was subcloned into an expression vector pET-29b. The recombinant collagenase was expressed in Escherichia coli BL2l(DE3) and partially purified by Hi-Trap affinity and Sephacryl S-100 size exclusion chromatographies. The recombinant enzyme was purified by 43.7-fold and the yield was 73%. SDS-PAGE revealed that the molecular weight of the enzyme was approximately 35 kDa. Substrate specificity study of the enzyme displayed that the enzyme showed the highest activity with the type I collagen and the synthetic peptide, Z-GPGGPA, indicating that the enzyme was indeed a collagenase. The enzyme showed broad pH optimum around pH 6-12 and was stable between pH 5.5 and 11.5. The optimum temperature for the type I collagen degradation was $35^{\circ}C$. The thermostability measurement of the enzyme indicated that the enzyme was stable up to $55^{\circ}C$, but the activity was diminished quickly above $60^{\circ}C.$

• The Korea Journal of Herbology
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• v.29 no.4
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• pp.21-27
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• 2014

Objectives : This experimental study was performed in order to investigate the antibacterial effect of bio-fermented Galla Rhois extract. Methods : The Galla Rhois extract was fermented by Streptococcus thermophilus, Saccharomyces cerevisiae and Lactobacillus delbrueckii, and their products was tested for antibacterial activity against six pathogenic microorganisms namely, Bacillus cereus, Bacillus subtilis, Staphylococcus aureus, Vibrio parahaemolyticus, Escherichia coli and Salmonella typhimurium by paper disc diffusion method. Results : The Galla Rhois fermented extract by Lactobacillus delbrueckii and Saccharomyces cerevisiae showed more effective antibacterial activity than not fermented extract against Bacillus subtilis and Vibrio parahaemolyticus. Antibacterial activity of fermented extract using especially Lactobacillus delbrueckii and Saccharomyces cerevisiae was proved that it was good with even 2 percents concentration. Antibacterial activity of Galla Rhois extract within pH 3 to pH 7 had been safe regardless of pH but low over pH 9. The growth of Bacillus cereus, Staphylococcus aureus, and Vibrio parahaemolyticus had a tendency to decrease depend on the increasing concentration of the extract. EtOEt, EtOAc and n-BuOH fractions of the Galla Rhois extract had a high level of antibacterial activity against Bacillus cereus, Bacillus subtilis, Staphylococcus aureus and Vibrio parahaemolyticus, respectively. Surprisingly, EtOAc fractions of the Galla Rhois extract showed higher antibacterial activity against Vibrio parahaemolyticus alone. And antibacterial activity against six pathogenic microorganisms had a tendency to increase depend on the increasing concentration of the fractions of the Galla Rhois extract. Conclusions : Bio-fermented Galla Rhois extract, efficiently inhibited the growth of Bacillus cereus and Vibrio parahaemolyticus.

• Fisheries and Aquatic Sciences
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• v.27 no.7
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• pp.434-446
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• 2024

The efficacy of Chromolaena odorata leaf flour components (CO) to increase the growth performance, non-specific immune activity, and resistance against Vibrio parahaemolyticus of whiteleg shrimp (Litopenaeus vannamei) culture was evaluated. To this purpose, whiteleg shrimp post larvae were fed on diets supplemented with 0 and a 1.5 g CO/kg diet for 45 days in a pond. After feeding trials, the shrimps were identified for their growth parameters, collected, and injected with V. parahaemolyticus, then their non-specific immune activity and resistance to V. parahaemolyticus was observed for 14 days. Findings showed that CO increased the average body weight (7.71 g, 37%), weight gain (7.69 g, 40%), and specific growth rate (13.23%/day, 5.7%) as compared to the control. In addition, CO supplementation also increases shrimp's hematologic and immune activity (total hemocyte counts [6.8 × 10⁷ CFU/mL, 242.9%], differential hemocyte counts [27%, 142.1%], and prophenoloxidase activity [0.085%, 566.7%]). Finally, shrimp resistance to V. parahaemolyticus infection also increased after CO supplementation, with survival rates of 73.33% as compared to 23.33% for the control. It suggests that C. odorata leaf flour component supplementation at an optimal dose in the diet may be an effective strategy to increase growth and resistance to bacterial disease with reduced mortality in shrimp farms.

• Journal of fish pathology
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• v.32 no.2
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• pp.97-104
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• 2019

The purpose of this study was to investigate the antimicrobial resistance and resistance transfer of Vibrio parahaemolyticus and Morganella morganii isolated from fish products purchased from fish markets in Yeosu April - December 2017. These bacteria were identified by biochemical test and PCR results, and the transfer of antimicrobial resistance was confirmed by the broth mating method. To isolate the transconjugants formed during conjugation, TSA medium containing 50 ㎍/ml of ampicillin (AMP), and 150 ㎍/ml of streptomycin (SM) or 30 ㎍/ml of oxytetracycline (OT) was used. M. morganii isolates showed low susceptibility to AMP, amoxicillin (AML), and colistin (CT), erythromycin, OT, and tetracycline, compared to V. parahaemolyticus resistance to AMP, AML, and CT. The conjugation of V. parahaemolyticus or M. morganii with Escherichia coli resulted in the separation of V. parahaemolyticus and M. morganii showing SM resistance as transconjugants. Meanwhile, Edwardsiella tarda transconjugants showing AMP and AML resistance were obtained from the broth mating of V. parahaemolyticus and E. tarda. But the transfer of the VPA0477 which is a β-lactamase gene of V. parahaemolyticus was not confirmed. These results suggest that resistance transfer between pathogenic bacteria is bidirectional and progresses in a wide variety of patterns.

• Korean Journal of Food Science and Technology
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• v.47 no.6
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• pp.719-724
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• 2015

The bactericidal effects of strongly acidic hypochlorous acid water (StAHA) and slightly acidic hypochlorous acid water (SlAHA) against Vibrio parahaemolyticus contaminated on surface of raw fish and shellfish were examined. V. parahaemolyticus contaminated with about 7.0 log CFU/g on the meat chunk of olive flounder (Paralichthys olivaceus), and yellow tail (Seriola quinqueradiata), and 4.0 log CFU/g on the shucked scallop (Patinopecten yessoensis) were not detected after washing with StAHA and SlAHA at a ratio of 30:1 on a sample weight basis. However, 1.0 log CFU/g of V. parahaemolyticus was survived on shucked oyster (Crassostrea gigas) under same treatment conditions. The bactericidal effects of acidic hypochlorous acid water against V. parahaemolyticus contaminated on surface of shucked oyster were not as effective as those against V. parahaemolyticus contaminated on surface of meat chunk of olive flounder, yellow tail, and shucked scallop. Such differences can be attributed to the complicated surface conformation of oyster.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.19 no.2
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• pp.136-140
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• 1986

The change of cell counts of Vibrio parahaemolyticus in fish muscle by the storage time and temperature was examined to get basic informations for precautionary steps against food poisoning of slices of raw fish (sashimi). There fore, we inoculated fish homogenate of oceanic bonito (Katsuwonus pelamis), yellow tail (Seriola quinqueradiata) with Kanagawa positive Vibrio parahaemolyticus and stored it at $30^{\circ}C,\;18^{\circ}C,\;4^{\circ}C\;and\;-20^{\circ}C$ for 24 hours. The number of the Vibrio parahaemolyticus upon fish homogenate stored at $30^{\circ}C\;and\;18^{\circ}C$ decreased for the first two hours and increased thereafter. When the fish homogenates inoculated with Vibrio parahaemolyticus at about $10^3$ per gram were stored at $18^{\circ}C\;and\;30^{\circ}C$ for 10 hours, the cell numbers increased about 10 times and 1,000 times initial cell numbers, respectively. The survival rate of Vibrio parahaemolyticus was about $20\%$, when the inoculated fish homogenates were stored at $-20^{\circ}C$ for 24 hours. Vibrio parahaemolyticus inoculated in fish homogenates was decreased by about $10\%$ of initial cell numbers by the storage at $4^{\circ}C$ for 4 hours and it was decreased by about $50\%$ after 24 hours storage of the samples at the same temperature. The decreasing rate of inoculated Vibrio parahaemolyticus in fresh fish muscle homogenate was higher than that in frozen fish muscle homogenate during the storage time at a refrigerator.