Abstract
The pathogenic species of genusVibrio cause vibriosis, one of the most prevalent diseases of maricultured animals and seafood consumers. Monitoring their kinetics in the chain of seafood production, processing and consumption is of great importance for food and mariculture safety. In order to enrichVibrio-representing 16S ribosomal RNA gene (rDNA) fragments and identify these bacteria further real-timely and synchronously among bacterial flora in the chain, a pair of primers that selectively amplifyVibrio 16S rDNA fragments were designed with their specificities and coverage testified in the analysis of seawaterVibrio community. The specificities and coverage of two primers, VF169 and VR744, were determined theoretically among bacterial 16S rDNAs available in GenBank by using BLAST program and practically by amplifying,Vibrio 16S rDNA fragments from seawater DNA. More than 88.3% of sequences in GenBank, which showed identical matches with VR744, belong toVibrio genus. A total of 33 clones were randomly selected and sequenced. All of the sequences showed their highest similarities to and clustered around those of diverse knownVibrio species. The primers designed are capable of retrieving a wide range ofVibrio 16S rDNA fragments specifically among bacterial flora in seawater, the most important natural environment of seafood cultivation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Actis, I. A., M. E. Tolmasky, and J. H. Crosa, 1999. Vibriosis.Fish. Dis. Disorders.,3: 523–557.
Austin, B., L. F. Stuckey, P. A. W. Robertson, I. Effendi, and D. R. W. Griffith, 1995. A probiotic strain ofVibrio alginolyticus effective in reducing diseases caused byAeromonas salmonicida, Vibrio anguillarum andVibrio ordalii.J. Fish Dis.,18: 93–96.
Baumann, P., A. L. Furniss, and J. V. Lee, 1984.Vibrio. In:Bergey's Manual of Systematic Bacteriology. Krieg, N. R., and J. G. Holt, eds., Williams & Wilkins, Baltimore, 518–544.
Bej, A. K., W. Y. Ng, S. Morgan, D. D. Jones, and M. H. Mahbubani, 1996. Detection of viableVibrio cholerae by reverse-transcriptase polymerase chain reaction (RT-PCR).Mol. Biotech.,5: 1–10.
David, M., and P. Herve, 2000. Molecular phylogeny: pitfalls and progress.Int. Microbiol.,3: 9–16.
Giovannoni, S., and M. Rappe, 2000. Evolution, diversity, and molecular ecology of marine prokaryotes. In:Microbial Ecology of the Oceans. Kirchman, D. L., ed., John Wiley & Sons, New York, 47–85.
Guschin, D. Y., B. K. Mobarry, D. Proudnikov, D. A. Stahl, B. E. Rittmann,et al., 1997. Oligonucleotide microchips as genosensors for determinative and environmental studies in microbiology.Appl. Environ. Microbiol.,63: 2397–2402.
Kita-Tsukamoto, K., H. Oyaizu, K. Nanba, and U. Simidu, 1993. Phylogenetic relationships of marine bacteria, mainly members of the family Vibrionaceae, determined on the basis of 16S rRNA sequences.Int. J. Syst. Bacteriol.,43: 8–19.
Kwok, S., S.-Y. Chang, J. J. Sninsky, and A. Wang, 1994. A guide to the design and use of mismatched and degenuste primers.PCR Method. Appl.,3: S39-S47.
Liu, J. Y., 1992. Characteristics of physical environment. In:Ecology and Living Resources of Jiaozhou Bay. Liu, J. Y., ed., Science Press, Beijing, 2–3.
MacDonell, M. T., and R. R. Colwell, 1985. Phylogeny of the Vibrionaceae and recommendation for two genus,Listonella andShewanella.Syst. Appl. Microbiol.,6: 171–182.
Marta, C. C., J. Joan, and R. B. Anicet, 2000. A selective medium and a specific probe for detection ofVibrio vulnificus.Appl. Environ. Microbiol.,66: 855–859.
Martin-Kearley, J., and J. A. Gow, 1994a. Numerical taxonomy of Vibrionaccae from Newfoundland coastal waters.Can. J. Microbiol. 40: 355–361.
Martin-Kearlcy, J., and J. A. Gow, 1994b. Numerical analysis and the application of random amplified polymorphic DNA polymerase chain reaction to the differentiation ofVibrio strains from a seasonally cold ocean.Can. J. Microbiol. 40: 446–455.
Muroga, K., 1995. Viral and bacterial diseases in larval and juvenile marine fish and shellfish-A review.Fish Pathol.,30: 71–85.
Olsen, G. J., C. R. Woese, and R. Overbeek, 1994. The winds of (evolutionary) change: breathing new life into microbiology.J. Bacteriol.,176: 1–6.
Ortigosa, M., E. Garay, and M. Pujalte, 1994. Numerical taxonomy of Vibrionaceac isolated from oysters and seawater along an annual cycle.Syst. Appl. Microbiol.,17: 216–225.
Polen-Fuller, M., 1991. A two-hour method for extraction of DNA from scawceds.Phycol. Newslett.,23: 2.
Sacchi, C. T., A. M. Whitney, L. W. Mayer, R. Morey, A. Steigerwalt,et al., 2002. Sequencing of 16S rRNA gene: A rapid tool for identification ofBacillus anthracie.Emerging Infect. Dis.,8: 1117–1123.
Saitou, N., and M. Nei, 1987. The neighbor-joining method: a method for reconstructing phylogenetic trees.Mol. Biol. Evol.,4: 406–425.
Smith, S. K., D. C. Sutton, J. A. Fuerst, and J. L. Reichelt, 1991. Evaluation of the genusListonella and reassignment ofListonella damsela (Loveet al.) MacDonell and Colwell to the genusPhotobacterium asPhotobacterium damsela comb. nov. with an emended description.Int. J. Syst. Bacteriol.,41: 529–534.
Sommer, R., and D. Tautz, 1989. Minimal homology requirements for PCR primers.Nucleic Acids Res.,17: 6749.
Stackebrandt, E., and B. M. Goebel, 1994. Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species in bacteriology.Int. J. Syst. Bacteriol.,44: 846–849.
Thompson, F. L., C. C. Thompson, B. Hoste, K. Vandemeulebroecke, M. Gullian,et al., 2003a.Vibrio fortis sp. nov. andVibrio hepatarius sp. nov., isolated from aquatic animals and the marine environment.Int. J. Syst. Evol. Microbiol.,53: 1495–1501.
Thompson, F. L., C. C. Thompson, Y. Li, B. Gomez-Gil, J. Vandenberghe,et al., 2003b.Vibrio kanaloae sp. nov.,Vibrio pomeroyi sp. nov. andVibrio chagasii sp. nov., from sea water and marine animals.Int. J. Syst. Evol. Microbiol.,53: 753–759.
Torres, L., S. Escobar, A. I. López, M. L. Marco, and V. Pobo, 2002. Wound infection due toVibrio vulnificus in Spain.Eur. J. Clin. Microbiol. & Infect. Dis.,21: 537–538.
Walter, E. H., P. K. Stacye, W. T. Mary, F. Peter, A. K. Charles,et al., 1991. Polymerase chain reaction identification ofVibrio vulnificus in artificially contaminated oysters.Appl. Environ. Microbiol.,57: 707–711.
Woese, C. R., and G. E. Fox, 1977. Phylogenetic structure of the prokaryotic domain: the primary kingdoms.Proc. Natl. Acad. Sci. USA,74: 5088–5090.
Xu, H., 1999. Methods for microbiology examination of vibriosis in aquacultrual animals. In:Diagnosis and Control of Bacterial Diseases in Penaeid Shrimp Hatcheries. Xu, H.,et al., eds., China Ocean Press, Beijing, 166–190.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yong, L., Guanpin, Y., Hualei, W. et al. Design ofVibrio 16S rRNA gene specific primers and their application in the analysis of seawaterVibrio community. J. Ocean Univ. China 5, 157–164 (2006). https://doi.org/10.1007/BF02919216
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02919216