• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.2
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• pp.185-190
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• 2003

The effects of pine needle extract and horseradish on the growth of Vibrio isolated from crab and flat fish were investigated. The isolated Vibrios were identified as Vibrio parahaemolyticus HY I and V. vulnificus FST I by Api 20E kit. The growth of V. parahaemolyticus HY 1, V. vulnificus FST I and V. perahaernolytich ATCC17802 were inhibited in tryptic soy broth (TSB) containing 1% pine needle ethanol extract. The growth of the Vibrios was more 2 log inhibited in TSB containing 1% pine needle extract and 1% horseradish than in TSB containing 1% horseradish alone. Viable cells of tile Vibrios were decreased more rapidly about 2~3 log in soysauce containing 1% pine needle extract and 1% of horseradish than in soysauce and in soysauce containing 1% horeseradish. Sensory quality of horseradish sauce containing 1% of pine needle extract was similar to that of horseradish sauce (p<0.05).

• Food Science and Biotechnology
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• v.15 no.6
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• pp.866-870
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• 2006

The maintenance of physiological activity during dilution is very critical for the accurate enumeration of Vibrio spp. in marine samples. We investigated the effect of various diluents on the recovery of Vibrio parahaemolyticus using the direct plate counting and most probable number (MPN) methods. The effects of NaCl (0.85 and 3%) and pH (from 6.6 to 7.4) in diluents based on distilled water or phosphate buffered saline (PBS) were evaluated with three V. parahaemolyticus strains. PBS-3% NaCl (pH 6.6), as opposed to PBS, was the most effective diluent at maintaining viable cell numbers up to 2 log CFU/g during dilution for direct plate counting using on thiosulfate citrate bile salts sucrose (TCBS) selective agar, as well as minimizing the difference in cell numbers between TCBS and non-selective nutrient agar. It also increased counts of V parahaemolyticus inoculated into oysters relative to PBS (p<0.01), suggesting that PBS-3% NaCl (PH 6.6) can reduce the problem of underestimating V. parhaemolyticus counts using PBS alone.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.55 no.6
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• pp.850-857
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• 2022

Seventy-five Vibrio parahaemolyticus isolates from the surface seawater of the Geum River Estuary area, on the west coast of Korea, were analyzed for the presence of virulence genes and susceptibility to 17 different antimicrobials. All 75 isolates were examined for the presence of two virulence genes (tdh or trh) using polymerase chain reaction; Only one of the isolates possessed the tdh or trh gene. According to the results of disk diffusion susceptibility tests, all of the strains were resistant to penicillin G, 92.0% were resistant to ampicillin, 82.7% were resistant to amoxicillin, 2.7% were resistant to ciprofloxacin, 2.7% were resistant to trimethoprim, 1.3% were resistant to cephalothin, and 1.3% were resistant to erythromycin. However, all of the strains were susceptible to amikacin, cefoxitin, chloramphenicol, gentamycin, kanamycin, nalidixic acid, nitrofurantoin, rifampin, streptomycin, and tetracycline. The average minimum inhibitory concentrations for ampicillin for V. parahaemolyticus was 557.6 ㎍/mL. These results not only provide novel insight into the necessity for seawater sanitation in Geum river estuary area, but they help reduce the risk of contamination of antimicrobial-resistant bacteria.

• Food Science and Biotechnology
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• v.15 no.4
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• pp.578-581
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• 2006

Fishery products were collected in seafood markets located on the southwestern coast of Korea between 2000 and 2004 and examined for the presence of Vibrio parahaemolyticus. This strain was detected in 138 of 843 samples (16.4%) that included dams, eels, crabs, octopuses, and cockles. The number of positive findings for V. parahaemolyticus among fishery products was the highest in dams at 23.6% followed by eels at 22.1%, crabs at 21.1%, octopuses at 18.0%, and cockles at 14.3%. V. parahaemolyticus was detected with overall frequencies of 15.3, 14.8, 13.8, 21.6, and 18.6% from 2000 to 2004, respectively. The monthly occurrence of the organism rapidly increased to over 20% between June and October. The monthly cases of food borne disease caused by V. parahaemolyticus in Korea over the last five years began to increase in August and reached its peak in September. However, the potential for outbreaks of food borne disease caused by V. parahaemolyticus was relatively minor between November and April. Consequently, this study shows that fishery products harvested from June to October must be handled sanitarily in Korea.

• Journal of Environmental Science International
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• v.5 no.5
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• pp.605-610
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• 1996

The bacteriophage from the fresh oyster, Crassostrea Virginica which is specific to the marine bacterium was isolated and characterized. Among the foci different vibrio species and the five different serotypes of Vibrio parahaemolyticus host strains tested, only two strains of the parahaemolyticus possessing K17, K52 antigens were highly sensitive to the phage. The size of the isolated plaque was 0.4mm and the electron microscopic head size of the isolated phage was about 67 nm long and 83 nm wide. PFU/ml was 1.25$\times$ $10^{11}$. The phase was sensitive to chloroform but resistant to acetone or methanol. The assay of the isolated phase nucleic acid was deoxyribonucleic acid. The restriction enzyme pattern showed 14 fragment from Hind III and 4 fragments from Eco R I. Two different antigenic groups showed-similar restriction enzyme patterns.

• Journal of Environmental Health Sciences
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• v.29 no.2
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• pp.94-102
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• 2003

This study was carried out to investigated the distribution and characteristics of Vibrio spp. isolated from fish and shellfish, seawater and brackish water samples collected from Pohang, Uljin, Yeongduk and Gyeongju in Gyeongbuk Province from April 2000 to October 2000. Total 155 strains of genus Vibrio were isolated from 439 collected samples, and numbers of isolated strains of V. parahaemolyticus and V. vulnificus were 140 and 15, respectively. The isolation rate from the samples collected in Pohang was the highest as 41.5% (76 strains), and wat the highest as 71.4% (30 strains) in brackish water, and was the highest as 55.7% (34 strains) in August. And the optimal pH, temperature, and NaCl concentration for growth of V. parahaemolyticus and V. cholerae were 8.0, 3$0^{\circ}C$ and 2.0％, respectively. In a resistance test for environmental factors, heat and cold resistants of V. parahaemolyticus were higher than those of V. vulnificus, withstanding for 15 minutes at 6$0^{\circ}C$ and 6 days at -18$^{\circ}C$. The pH range for existence of V. parahaemolyticus and V. vulnificus were 4.5~l1.0 and 4.5~10.0, showing the similar resistance to pH. V. parahaemolyticus and V. vulnificus could grow in media containing up to 10.0% and 7.0% NaCl, respectively, Salt-tolerance of V. parahaemolyticus was higher than that of V. vulnificus. In an antibiotics sensitivity test against 16 strains of V. parahaemolyticus, twelve strains were resistant to ampicillin, eight strains were resistant to cephalothin. one strain was resistant to streptomycin, and one strain was resistant to ticarcillin.

• Journal of Life Science
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• v.14 no.2
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• pp.362-367
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• 2004

For better understanding of the host infection mechanism of Vibrio, a Vibrio parahaemolyticus collagenase mutant was generated by insertional inactivation of a vppC gene encoding extracellular collagenase. A recombinant DNA containing vppC::nptII was cloned into a suicide plasmid pDMS197, resulted in pVCM03. The recombinant suicide plasmid pVCM03 contained in E. coli $\chi$7213 was transferred to a wild-type V. parahaemolyticus 04 through conjugation. The recombinant vppC::nptII DNA in pVCM03 was exchanged with wild-type allele by homologous recombination resulting vppC mutant, V. parahaemolyticus CM. The mutant was selected and screened on TCBS media containing 10% sucrose and kanamycin. The mutation by allele exchange was confirmed with the comparison of the size of DNAs amplified by PCR. V. parahaemolyticus CM showed at least 4-fold less collagen-degrading activity than those of wild-type, and the mutant exhibited less cytotoxicity than that of wild-type in MTT assay.

• 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.31 no.12
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• pp.1672-1683
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• 2021

Vibrio parahaemolyticus is recognized as one of the most important foodborne pathogens responsible for gastroenteritis in humans. The bla_CARB-17 gene is an intrinsic β-lactamase gene and a novel species-specific genetic marker of V. parahaemolyticus. In this study, a loop-mediated isothermal amplification (LAMP) assay combined with a lateral flow dipstick (LFD) was developed targeting this bla_CARB-17 gene. The specificity of LAMP-LFD was ascertained by detecting V. parahaemolyticus ATCC 17802 and seven other non-V. parahaemolyticus strains. Finally, the practicability of LAMP-LFD was confirmed by detection with V. parahaemolyticus-contaminated samples and natural food samples. The results showed that the optimized reaction parameters of LAMP are as follows: 2.4 mmol/l Mg²⁺, 0.96 mmol/l dNTPs, 4.8 U Bst DNA polymerase, and an 8:1 ratio of inner primer to outer primer, at 63℃ for 40 min. The optimized reaction time of the LFD assay is 60 min. Cross-reactivity analysis with the seven non-V. parahaemolyticus strains showed that LAMP-LFD was exclusively specific for V. parahaemolyticus. The detection limit of LAMP-LFD for V. parahaemolyticus genomic DNA was 2.1 × 10^-4 ng/μl, corresponding to 630 fg/reaction and displaying a sensitivity that is 100-fold higher than that of conventional PCR. LAMP-LFD in a spiking study revealed a detection limit of approximately 6 CFU/ml, which was similar with conventional PCR. The developed LAMP-LFD specifically identified the 10 V. parahaemolyticus isolates from 30 seafood samples, suggesting that this LAMP-LFD may be a suitable diagnostic method for detecting V. parahaemolyticus in aquatic foods.

• 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.