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Isolation, Detection and Genomic Differentiation of Vibrio cholerae and Vibrio parahaemolyticus in Bachok, Kelantan

Vimala, B., and Thong, Kwai Lin, and Chong, V. C., (2010) Isolation, Detection and Genomic Differentiation of Vibrio cholerae and Vibrio parahaemolyticus in Bachok, Kelantan. Malaysian Journal of Science, 29 (Special Issue). pp. 1-10. ISSN 13943065

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Affiliations

University of Malaya, Faculty of Science, Institute of Biological Sciences
University of Malaya, Faculty of Science, Institute of Biological Sciences
University of Malaya, Faculty of Science, Institute of Biological Sciences

Abstract

The objective of this study was to determine the genetic diversity and virulence factors of Vibrio cholerae and V. parahaemolyticus from selected aquatic environments in Bachok, Kelantan, Malaysia. Out of 50 water samples, 33 were confirmed with the presence of presumptive V. cholerae and V. parahaemolyticus based on biochemical tests. Primers targeting toxR gene specific for V. cholerae (800bp) and V. parahaemolyticus (368bp), tdh (251bp) and trh (250bp) virulence genes were used in the PCR. Results of PCR showed that only 25 isolates of V. parahaemolyticus and 3 of V. cholerae had the toxR gene. None of the V. parahaemolyticus isolates had the trh and tdh genes. Repetitive extragenic palindromic PCR (REP-PCR) was used to determine the relatedness of these strains at the genomic level. Cluster analysis for REP-PCR showed 2 major clusters A-B, with several isolates from different locations grouped together indicating the geographical relatedness due to interconnected waterways. REPPCR showed that the species of Vibrios are genetically diverse.

Objektif kajian ini adalah untuk menentukan diversiti gen and faktor-faktor virulen Vibrio cholerae dan Vibrio parahaemolyticus yang diperolehi di sekitar perairan Bachok, Kelantan. Sejumlah 33 daripada 50 sampel air yang diuji melalui kaedah biokimia didapati positif untuk V. cholerae dan V. parahaemolyticus. Bagi amplifikasi PCR, primer toxR yang spesifik kepada V. cholerae (800bp) dan V. parahaemolyticus (368bp) serta primer-primer untuk mengesan gen-gen virulen seperti tdh (251bp) and trh (250bp) juga digunakan. Hasil PCR menunjukkan hanya 25 strain V. parahaemolyticus dan 3 strain V. cholerae mempunyai gen toxR. Tiada gen virulen dikesan pada mana-mana strain V. parahaemolyticus. REP-PCR diaplikasi untuk menentukan hubungan antara strain-strain yang telah diasingkan dalam kajian ini. Analisis dendrogram bagi REP-PCR menunjukkan 2 kumpulan utama, A-B. Strain-strain yang berasal dari lokasi yang berbeza dikumpulkan bersama. Ini mungkin disebabkan oleh faktor geografi di mana sistem perairan yang berkaitan. Keputusan REP-PCR menunjukkan variasi genetik yang nyata di dalam spesies Vibrio.

Item Type:Journal
Additional Information:The authors thank University of Malaya for the short term research grant (FS318/2008A) for this work. The authors would like to thank Tim Chuan Seet, Rathi, and Lim Sin Yun for technical assistance during the field sampling in Bachok, Kelantan.
Keywords:Vibrios, toxR , tdh , trh , REP-PCR
Subjects:Q Science, Computer Science
ID Code:10890

1. Colwell, R.R. (2004). Infectious disease and environment: cholera as a paradigm for waterborne disease. Int Microbiol 7: 285–289.

2. Summer, J.D.A., Osaka, K., Karunasager, I., Walkderhaug, M. and Bowers, J. (2001). Hazard identification, exposure assessment, and hazard characterization of Vibrio sp. in seafood. In Joint FAO/WHO Activities on Risk Assessment of Microbiological Hazards in Foods, pp. 1–105.

3. Thompson, F.L., Iida, T. and Swings, J. (2004). Biodiversity of vibrios. Microbiol Mol Biol Rev 68:403–431

4. Yuherman, Son R., Rusul G., Lum K.Y. and Jamakhair (2000). Genetic relatedness of Vibrio Cholerae O1 and non-O1 by ERIC Polymerase Chain Reaction and Pulse Field Gel Electrophoresis. Pakistan J Biol Sci 3(4): 621-622.

5. Chen, C. H., Shimada, T., Elhadi, N., Radu, S. and Nishibuchi, M., (2004). Phenotypic and Genotypic Characteristics and Epidemiological Significance of ctx+ Strains of Vibrio cholerae Isolated from Seafood in Malaysia. Appl Environ Microbiol 70: 19641972.

6. Tanil G.B, Son R., Nishibuchi M., Raha AR., Napis S., Maurice L. and Gunsalam J.W. (2005) .Characterization of Vibrio parahaemolyticus isolated from coastal seawater in peninsular Malaysia. Southeast Asian J Trop Med Publ Health 36(4):940-5

7. Lesley M.B., Son R., Abdul R.B., Raha A.R., Suhaini N., Cheah Y.K., Chandrika M., Yousr A.H., Tunung R. and Nishibuchi M. (2005). Random Amplified Polymorphic DNA-PCR

typing of Vibrio parahaemolyticus isolated from local cockles (Anadara granosa). Am J Immunol 1(1): 31-36

8. Vadivelu, J., Iyer, L., Kshatriya, B.M. and Puthucheary, S.D. (2000). Molecular evidence of clonality amongst Vibrio cholerae O1 biotype El Tor during an outbreak in Malaysia. Epidemiol Infect 124: 25-30

9. Iyer, L. and Vadivelu, J. (2006). Molecular analysis of Vibrio cholerae strains isolated in Malaysia: Public health implications. Asia Pac J Publ Health 18(3): 33-41

10. Okuda, J., Ishibashi, M., Hayakawa, E., Nishino, T., Takeda, Y., Mukhopadhyay, A. K., et.al. (1997). Emergence of a unique O3:K6 clone of Vibrio parahaemolyticus in Calcutta, India, and isolation of strains from the same clonal group from Southeast Asian travelers arriving in Japan. J Clin Microbiol 35:3150–3155

11. Dileep, V., Kumar, H.S., Kumar, Y., Nishibuchi, M. and Karunasagar, I. (2003). Application of polymerase chain reaction for detection of Vibrio parahaemolyticus associated with tropical seafood and coastal environment. Lett Appl Microbiol 36: 423–427

12. Chiou, C.S., Hsu, S.Y., Chiu, S.I., Wang, T.K. and Chao, C.S. (2000). Vibrio parahaemolyticus serovar O3:K6 as cause of unusually high incidence of food-borne disease outbreaks in Taiwan from 1996 to 1999. J Clin Microbiol 38: 4621–4625

13. Robert-Pillot, A., Guénolé, A., Lesne, J., Delesmont, R., Fournier, J.M. and Quilici, M.L. (2004). Occurrence of the tdh and trh genes in Vibrio parahaemolyticus isolates from waters and raw shellfish collected in two French coastal areas and from seafood imported into France. International J Food Microbiol 91: 319–325

14. Khan, A.A., McCarthy S., Wang R.F. and Cerniglia C.E. (2002). Characterization of United States outbreak isolates of Vibrio parahaemolyticus using enterobacterial repetitive intergenic consensus (ERIC) PCR and development of a rapid PCR method for detection of O3:K6 isolates. FEMS Microbiol Lett 206: 209-214

15. Centers for Disease Control and Prevention (CDC), (1998). Outbreak of Vibrio parahaemolyticus infections association with eating raw oysters - Pacific Northwest, 1997. Morb Mortal Wkly Rep 47:457–462

16. Centers for Disease Control and Prevention (CDC), (1999). Outbreak of Vibrio parahaemolyticus infection associated with eating raw oysters and clams harvested from Long Island Sound—Connecticut, New Jersey and New York, 1998. Morb Mortal Wkly Rep 48:48–51

17. Elhadi, N., Radu, S., Chen C.H. and Nishibuchi, M. (2004). Prevalence of potentially pathogenic Vibrio species in the seafood marketed in Malaysia. J Food Prot 67: 1469-1475

18. Miyamoto, Y., Kato, T., Obara, Y., Akiyama, S., Takizawa, K. and Yamai, S. (1969). In vitro hemolytic characteristic of Vibrio parahaemolyticus: its close correlation with human pathogenicity. J Bacteriol 100:1147–

1149.

19. Honda, T. and Iida, T. (1993).The pathogenicity of Vibrio parahaemolyticus and the role of thermostable direct haemolysin and related haemolysins. Rev Med Microbiol 4: 106–113.

20. Hervio-Heath, D., Colwell, R.R., Derrien, A., Robert-Pillot, A., Fournier, J.M. and Pommepuy, M. (2002). Occurrence of pathogenic vibrios in coastal areas of France. J Appl Microbiol 92:1123–1135

21. Nishibuchi, M. and Kaper, J.B (1995) Thermostable direct haemolysin gene of Vibrio parahaemolyticus: a virulence gene acquired by a marine bacterium. Infect Immun 63:2093–2099

22. Joseph, S.W., Colwell, R.R. and Kaper, J.B. (1982). Vibrio parahaemolyticus and related halophilic vibrios. Crit Rev Microbiol 10:77–124

23. Bej, A.K., Patterson, D.P., Brasher, C.W., Vickery, M.C.L., Jones, D.D. and Kaysner, C.A. (1999). Detection of total and hemolysinproducing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh, and

trh. J Microbiol Methods 36: 215–225

24. Kim, Y.B., 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 tox R gene. J Clin Microbiol 37: 1173–1177

25. Wong, H. C., and Lin C.H. (2001). Evaluation of typing of Vibrio parahaemolyticus by three PCR methods using specific primers. J Clin Microbiol 39:4233–4240

26. Hara-Kudo Y., Nishina T., Nakagawa H., Konuma H., Hasegawa J. and Kumagai S. (2001). Improved method for detection of Vibrio parahaemolyticus in seafood. Appl Environ Microbiol 67:5819–5823

27. Tada, J., Ohashi, T., Nishimura, N., Shirasaki, Y., Ozaki, H., Fukushima, S., Takano, J., Nishibuchi, M. and Takeda, Y. (1992). Detection of the thermostable direct hemolysin gene (tdh) and the thermostable direct hemolysin-related hemolysin gene (trh) of Vibrio parahaemolyticus by polymerase chain reaction. Mol Cell Probes 6: 477-487

28. Rivera, I.N.G., Chun, J., Huq, A., Sack, R.B. and Colwell, R.R. (2001). Genotypes associated with virulence in environmental isolates of Vibrio cholerae. Appl Environ Microbiol 67: 2421–2429

29. Choopun, N., Louis, V. Huq, A. and Colwell R.R. (2002). Simple procedure for rapid identification of Vibrio cholerae from the aquatic environment. Appl Environ Microbiol 68:995-998

30. Colwell, R.R. and Huq, A. (1994). Vibrio cholerae and Cholera: Molecular to Global Perspectives. Washington, DC: ASM Press, pp.117–135.

31. Oliver, J. and Koper, J. (1997). Vibrio species In: Food Microbiology: Fundamentals and Frontiers. Washington DC: Editorial ASM Press, pp. 228-264

32. Tamrakar, A.K., Goel, A.K., Kamboj, D.V. and Singh, L. (2006). Surveillance methodology for Vibrio cholerae in environmental samples. Int J Environ Health Res 16(4):305-12

33. Cook, D. W., Bowers, J. C. and DePaola, A.(2002). Density of total and pathogenic (tdh+) Vibrio parahaemolyticus in Atlantic and Gulf coast molluscan shellfish at harvest. J Food Prot 65:1873–1880

34. Kiiyukia, C., Venkateswaran, K., Navarro, I. M., Nakano, H., Kawakami, H. and Hashimoto, H. (1989). Seasonal distribution of Vibrio parahaemolyticus serotypes along the oyster beds in Hiroshima coast. Journal of Faculty of Applied Biological Science, Hiroshima University, 28:49–61

35. Ogawa, H., Tokunou, H., Kishimoto, T., Fukuda, S., Umemura, K., and Takata, M. (1989). Ecology of Vibrio parahaemolyticus in Hiroshima Bay. Hiroshima. J Vet Med 4: 47–57

36. Faruque, S.M., Albert, M.J. and Mekalanos, J.J. (1998). Epidemiology,genetics and ecology of toxigenic Vibrio cholerae. Microbiol Mol Biol Rev 62: 1301–1314

37. Olive, D. M. and Bean, P. (1999). Principles and applications of methods for DNA based typing of microbial organisms. J Clin Microbiol 37:1661–1669

38. Navia, M.M, Capitano, L., Ruiz, J., Vargas M., Urassa H., Schellemberg D., Gascon J. and Vila, J. (1999). Typing and characterization of mechanisms of resistance of Shigella spp. isolated from faeces of children under 5 years of age from Ifakara, Tanzania. J Clin Microbiol 37:

3113–3117

39. Szczuka, E., and Kaznowski, A. (2004). Typing of clinical and environmental Aeromonas sp. strains by random amplified polymorphic DNA PCR, repetitive extragenic palindromic PCR, and enterobacterial repetitive intergenic consensus sequence PCR. J Clin Microbiol 42:220-228

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