Korean Journal of Fisheries and Aquatic Sciences (한국수산과학회지)

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Quantification of Vibrio parahaemolyticus Using a Most Probable Number-Polymerase Chain Reaction Assay Targeting the H-NS gene

MPN 및 H-NS 유전자를 표적으로 하는 PCR assay를 병용한 장염비브리오(Vibrio parahaemolyticus)의 정량

  • Kim, Tae-Ok (Department of Food Science and Biotechnology, Kunsan National University) ;
  • Park, Kwon-Sam (Department of Food Science and Biotechnology, Kunsan National University)
  • 김태옥 (군산대학교 식품생명공학과) ;
  • 박권삼 (군산대학교 식품생명공학과)
  • Received : 2014.09.26
  • Accepted : 2014.10.08
  • Published : 2014.10.31
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Abstract

We applied a combination of most probable number-polymerase chain reaction (MPN-PCR) methods using a PCR procedure targeting the H-NS (VP1133) gene to detect Vibrio parahaemolyticus presence and density in seawater as well as within short-necked clam Ruditapes philippinarum tissues collected from Gomso Bay, Korea. In 30 seawater samples, V. parahaemolyticus levels ranged from less than 1.8 to $1.1{\times}10^3MPN/100mL$, and samples from August showed higher than those from other months. Furthermore, the levels of V. parahaemolyticus in six short-necked clam samples ranged from $7.8{\times}10^2$ to $2.1{\times}10^3MPN/100g$, approximately 2.5 times higher than in seawater samples from the corresponding month. Our results provide data on V. parahaemolyticus contamination in seawater and short-necked clam tissues, and help to improve quantitative methods of assessing V. parahaemolytcius levels.

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References

  1. A.P.H.A. 1970. Recommended procedures for the examination of seawater and shellfish. 4th Ed. American Public Health Association, Washington DC, U.S.A, 1-47.
  2. Bej AK, Patterson DP, Brasher CW, Vickery MC, Jones DD and Kaysner CA. 1999. Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh and trh. J Microbiol Meth 36, 215-225. https://doi.org/10.1016/S0167-7012(99)00037-8
  3. Blake PA, Weaver RE and Hollis DG. 1980. Diseases of humans (other than cholera) caused by vibrios. Annu Rev Microbiol 34, 341-367. https://doi.org/10.1146/annurev.mi.34.100180.002013
  4. Cantet F, Hervio-Heath D, Caro A, Le Mennec C, Monteil C, Quemere C, Jolivet-Gougeon A, Colwell RR and Monfort P. 2013. Quantification of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae in French Mediterranean coastal lagoons. Res Microbiol 164, 867-874. http://dx.doi.org/10.1016/j.resmic.2013.06.005.
  5. Chen M, Guo D, Wong HC, Zhang X, Liu F, Chen H, Chen M, Liu B, Wang L, Wu F and Feng L. 2012. Development of Oserogroup specific PCR assay for detection and identification of Vibrio parahaemolyticus. Int J Food Microbiol 159, 122-129. http://dx.doi.org/10.1016/j.ijfoodmicro.2012.08.012.
  6. Copin S, Robert-Pillot A, Malle P, Quilici ML and Gay M. 2012. Evaluation of most-probable-number-PCR method with internal amplification control for the counting of total and pathogenic Vibrio parahaemolyticus in frozen shrimps. J Food Prot 75, 150-153. http://dx.doi.org/10.4315/0362-028X.JFP-11-165.
  7. DePaola A, Hopkins LH, Peeler JT, Wentz B and McPhearson RM. 1990. Incidence of Vibrio parahaemolyticus in U.S. coastal waters and oysters. Appl Environ Microbiol 56, 2299-2302.
  8. Fang FC and Rimsky S. 2008. New insights into transcriptional regulation by H-NS. Curr Opin Microbiol 11, 113-120. http://dx.doi.org/10.1016/j.mib.2008.02.011.
  9. Honda T and Iida T. 1993. The pathogenicity of Vibrio parahaemolyticus and the role of the thermostable direct haemolysin and related haemolysins. Rev Med Microbiol 4, 106-113. https://doi.org/10.1097/00013542-199304000-00006
  10. Hossain MT, Kim EY, Kim YR, Kim DG and Kong IS. 2011. Application of groEL gene for the species-specific detec-tion of Vibrio parahaemolyticus by PCR. Lett Appl Microbiol 54, 67-72. http://dx.doi.org/10.1111/j.1472-765X.2011.03174.x.
  11. Kaneko T and Colwell RR. 1975. Adsorption of Vibrio parahaemolyticus onto chitin and copepods. Appl Microbiol 29, 269-274.
  12. Kim YB, Okuda J, Matsumoto C, Takahashi N, Hashimoto S and Nishibuchi M. 1999. Identification of Vibrio parahaemolyticus strains at the species level by PCR targeted to the toxR gene. J Clin Microbiol 37, 1173-1177.
  13. Makino K, Oshima K, Kurokawa K, et al., 2003. Genome sequence of Vibrio parahaemolyticus: a pathogenic mechanism distinct from that of V. cholerae. Lancet 361, 743-749. https://doi.org/10.1016/S0140-6736(03)12659-1
  14. Ministry of Food and Drug Safety (MFDS). 2014. Korea food code. Retrieved from http://www.mfds.go.kr/e-stat/index.do?nMenuCode=20 on September 15.
  15. No AR, Okada K, Kogure K and Park KS. 2011. Rapid detection of Vibrio parahaemolyticus by PCR targeted to the histone-like nucleoid structure (H-NS) gene and its genetic characterization. Lett Appl Microbiol 53, 127-133. http://dx.doi.org/10.1111/j.1472-765X.2011.03072.x.
  16. Park KS, Arita M, Iida T and Honda T. 2005. vpaH, a gene encoding a novel Histone-like nucleoid structure-like protein that was possibly horizontally acquired, regulates the biogenesis of lateral flagella in trh-positive Vibrio parahaemolyticus TH3996. Infect Immun 73, 5754-5761. https://doi.org/10.1128/IAI.73.9.5754-5761.2005
  17. Sakazaki R, Tamura K, Kato T, Obara Y and Yamai S. 1968. Studies on the enteropathogenic, facultatively halophilic bacterium, Vibrio parahaemolyticus. 3. Enteropathogenicity. Jpn J Med Sci Biol 21, 325-331. https://doi.org/10.7883/yoken1952.21.325
  18. Shirai H, Ito H, Hirayama T, Nakamoto Y, Nakabayashi N, Kumagai K, Takeda Y and Nishibuchi M. 1990. Molecular epidemiological evidence for association of thermostable direct hemolysin (TDH) and TDH-related hemolysin of Vibrio parahaemolyticus with gastroenteritis. Infect Immun 58, 3568-3573.
  19. Su YC, Duan J and Wu WH. 2005. Selectivity and specificity of a chromogenic medium for detecting Vibrio parahaemolyticus. J Food Prot 68, 1454-1456. https://doi.org/10.4315/0362-028X-68.7.1454
  20. Venkateswaran K, Dohmoto N and Harayama S. 1998. Cloning and nucleotide sequence of the gyrB gene of Vibrio parahaemolyticus and its application in detection of this pathogen in shrimp. Appl Environ Microbiol 64, 681-687.
  21. Wang R, Huang J, Zhang W, Lin G, Lian J, Jiang L, Lin H, Wang S and Wang S. 2011. Detection and identification of Vibrio parahaemolyticus by multiplex PCR and DNA-DNA hybridization on a microarray. J Genet Genomics 38, 129-135. http://dx.doi.org/10.1016/j.jgg.2011.02.002.
  22. Wei S, Zhao H, Xian Y, Hussain MA and Wu X. 2014. Multiplex PCR assays for the detection of Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae with an internal amplification control. Diagn Microbiol Infect Dis 79, 115-118. http://dx.doi.org/10.1016/j.diagmicrobio.2014.03.012.
  23. Yu S, Chen W, Wang D, He X, Zhu X and Shi X. 2010. Speciesspecific PCR detection of the food-borne pathogen Vibrio parahaemolyticus using the irgB gene identified by comparative genomic analysis. FEMS Microbiol Lett 307, 65-71. http://dx.doi.org/10.1111/j.1574-6968.2010.01952.x.