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. 1997 Jul;63(7):2600–2606. doi: 10.1128/aem.63.7.2600-2606.1997

Intraspecific Differentiation of Vibrio vulnificus Biotypes by Amplified Fragment Length Polymorphism and Ribotyping

C R Arias, L Verdonck, J Swings, E Garay, R Aznar
PMCID: PMC1389195  PMID: 16535640

Abstract

The intraspecific genomic relatedness of 80 Vibrio vulnificus isolates, 44 of biotype 1 and 36 of biotype 2, from different geographic origins and sources was evaluated by ribotyping and AFLP (amplified fragment length polymorphism) fingerprinting. Ribopatterns of DNAs digested with KpnI and hybridized with an oligonucleotide complementary to a highly conserved sequence in the 23S rRNA gene revealed up to 19 ribotypes in the species, which were different for the two biotypes. Sixteen different ribotypes were found within biotype 1 strains from clinical and environmental sources, and only three, recovered mainly from diseased eels, were found within biotype 2. Within this biotype, 96% of the strains showed the same ribopattern. The closest similarity was shown by the strains coming from the same eel farm, irrespectively of biotype. AFLP fingerprints obtained by selective PCR amplification of HindIII-TaqI double-restricted DNA fragments exhibited a strain-specific pattern which allowed the finest differentiation of subgroups within the eel-pathogenic isolates sharing the same ribopattern. Both techniques revealed good genetic markers for intraspecific differentiation of V. vulnificus. Ribotyping clearly separated the eel-pathogenic strains from the clinical and environmental isolates, whereas AFLP enabled the monitoring of individual strains and therefore constitutes one of the most discriminative tools for epidemiological and ecological studies.

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Selected References

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  1. Amaro C., Biosca E. G., Fouz B., Alcaide E., Esteve C. Evidence that water transmits Vibrio vulnificus biotype 2 infections to eels. Appl Environ Microbiol. 1995 Mar;61(3):1133–1137. doi: 10.1128/aem.61.3.1133-1137.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Amaro C., Biosca E. G., Fouz B., Garay E. Electrophoretic analysis of heterogeneous lipopolysaccharides from various strains of Vibrio vulnificus biotypes 1 and 2 by silver staining and immunoblotting. Curr Microbiol. 1992 Aug;25(2):99–104. doi: 10.1007/BF01570967. [DOI] [PubMed] [Google Scholar]
  3. Amaro C., Biosca E. G., Fouz B., Toranzo A. E., Garay E. Role of iron, capsule, and toxins in the pathogenicity of Vibrio vulnificus biotype 2 for mice. Infect Immun. 1994 Feb;62(2):759–763. doi: 10.1128/iai.62.2.759-763.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Amaro C., Biosca E. G. Vibrio vulnificus biotype 2, pathogenic for eels, is also an opportunistic pathogen for humans. Appl Environ Microbiol. 1996 Apr;62(4):1454–1457. doi: 10.1128/aem.62.4.1454-1457.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Biosca E. G., Fouz B., Alcaide E., Amaro C. Siderophore-mediated iron acquisition mechanisms in Vibrio vulnificus biotype 2. Appl Environ Microbiol. 1996 Mar;62(3):928–935. doi: 10.1128/aem.62.3.928-935.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Biosca E. G., Garay E., Toranzo A. E., Amaro C. Comparison of outer membrane protein profiles of Vibrio vulnificus biotypes 1 and 2. FEMS Microbiol Lett. 1993 Mar 1;107(2-3):217–222. doi: 10.1111/j.1574-6968.1993.tb06033.x. [DOI] [PubMed] [Google Scholar]
  7. Biosca E. G., Oliver J. D., Amaro C. Phenotypic characterization of Vibrio vulnificus biotype 2, a lipopolysaccharide-based homogeneous O serogroup within Vibrio vulnificus. Appl Environ Microbiol. 1996 Mar;62(3):918–927. doi: 10.1128/aem.62.3.918-927.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Buchrieser C., Gangar V. V., Murphree R. L., Tamplin M. L., Kaspar C. W. Multiple Vibrio vulnificus strains in oysters as demonstrated by clamped homogeneous electric field gel electrophoresis. Appl Environ Microbiol. 1995 Mar;61(3):1163–1168. doi: 10.1128/aem.61.3.1163-1168.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dalsgaard A., Echeverria P., Larsen J. L., Siebeling R., Serichantalergs O., Huss H. H. Application of ribotyping for differentiating Vibrio cholerae non-O1 isolated from shrimp farms in Thailand. Appl Environ Microbiol. 1995 Jan;61(1):245–251. doi: 10.1128/aem.61.1.245-251.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fouz B., Mazoy R., Lemos M. L., del Olmo M. J., Amaro C. Utilization of hemin and hemoglobin by Vibrio vulnificus biotype 2. Appl Environ Microbiol. 1996 Aug;62(8):2806–2810. doi: 10.1128/aem.62.8.2806-2810.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grattard F., Pozzetto B., Ros A., Gaudin O. G. Differentiation of Pseudomonas aeruginosa strains by ribotyping: high discriminatory power by using a single restriction endonuclease. J Med Microbiol. 1994 Apr;40(4):275–281. doi: 10.1099/00222615-40-4-275. [DOI] [PubMed] [Google Scholar]
  12. Hor L.-I., Gao C.-T., Wan L. Isolation and Characterization of Vibrio vulnificus Inhabiting the Marine Environment of the Southwestern Area of Taiwan. J Biomed Sci. 1995 Oct;2(4):384–389. doi: 10.1007/BF02255226. [DOI] [PubMed] [Google Scholar]
  13. Huys G., Coopman R., Janssen P., Kersters K. High-resolution genotypic analysis of the genus Aeromonas by AFLP fingerprinting. Int J Syst Bacteriol. 1996 Apr;46(2):572–580. doi: 10.1099/00207713-46-2-572. [DOI] [PubMed] [Google Scholar]
  14. Janssen P., Coopman R., Huys G., Swings J., Bleeker M., Vos P., Zabeau M., Kersters K. Evaluation of the DNA fingerprinting method AFLP as an new tool in bacterial taxonomy. Microbiology. 1996 Jul;142(Pt 7):1881–1893. doi: 10.1099/13500872-142-7-1881. [DOI] [PubMed] [Google Scholar]
  15. Kaysner C. A., Abeyta C., Jr, Wekell M. M., DePaola A., Jr, Stott R. F., Leitch J. M. Virulent strains of Vibrio vulnificus isolated from estuaries of the United States West Coast. Appl Environ Microbiol. 1987 Jun;53(6):1349–1351. doi: 10.1128/aem.53.6.1349-1351.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kreger A. S., Gray L. D., Testa J. Protection of mice against Vibrio vulnificus disease by vaccination with surface antigen preparations and anti-surface antigen antisera. Infect Immun. 1984 Sep;45(3):537–543. doi: 10.1128/iai.45.3.537-543.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kreger A., DeChatelet L., Shirley P. Interaction of Vibrio vulnificus with human polymorphonuclear leukocytes: association of virulence with resistance to phagocytosis. J Infect Dis. 1981 Sep;144(3):244–248. doi: 10.1093/infdis/144.3.244. [DOI] [PubMed] [Google Scholar]
  18. Oliver J. D., Warner R. A., Cleland D. R. Distribution and ecology of Vibrio vulnificus and other lactose-fermenting marine vibrios in coastal waters of the southeastern United States. Appl Environ Microbiol. 1982 Dec;44(6):1404–1414. doi: 10.1128/aem.44.6.1404-1414.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Oliver J. D., Warner R. A., Cleland D. R. Distribution of Vibrio vulnificus and other lactose-fermenting vibrios in the marine environment. Appl Environ Microbiol. 1983 Mar;45(3):985–998. doi: 10.1128/aem.45.3.985-998.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Olsen J. E., Larsen J. L. Ribotypes and plasmid contents of Vibrio anguillarum strains in relation to serovar. Appl Environ Microbiol. 1993 Nov;59(11):3863–3870. doi: 10.1128/aem.59.11.3863-3870.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pedersen K., Larsen J. L. Evidence for the existence of distinct populations of Vibrio anguillarum serogroup O1 based on plasmid contents and ribotypes. Appl Environ Microbiol. 1995 Jun;61(6):2292–2296. doi: 10.1128/aem.61.6.2292-2296.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Salvi R. J., Ahroon W., Saunders S. S., Arnold S. A. Evoked potentials: computer-automated threshold-tracking procedure using an objective detection criterion. Ear Hear. 1987 Jun;8(3):151–156. [PubMed] [Google Scholar]
  23. Skov M. N., Pedersen K., Larsen J. L. Comparison of Pulsed-Field Gel Electrophoresis, Ribotyping, and Plasmid Profiling for Typing of Vibrio anguillarum Serovar O1. Appl Environ Microbiol. 1995 Apr;61(4):1540–1545. doi: 10.1128/aem.61.4.1540-1545.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tamplin M. L., Jackson J. K., Buchrieser C., Murphree R. L., Portier K. M., Gangar V., Miller L. G., Kaspar C. W. Pulsed-field gel electrophoresis and ribotype profiles of clinical and environmental Vibrio vulnificus isolates. Appl Environ Microbiol. 1996 Oct;62(10):3572–3580. doi: 10.1128/aem.62.10.3572-3580.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tison D. L., Nishibuchi M., Greenwood J. D., Seidler R. J. Vibrio vulnificus biogroup 2: new biogroup pathogenic for eels. Appl Environ Microbiol. 1982 Sep;44(3):640–646. doi: 10.1128/aem.44.3.640-646.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Veenstra J., Rietra P. J., Stoutenbeek C. P., Coster J. M., de Gier H. H., Dirks-Go S. Infection by an indole-negative variant of Vibrio vulnificus transmitted by eels. J Infect Dis. 1992 Jul;166(1):209–210. doi: 10.1093/infdis/166.1.209. [DOI] [PubMed] [Google Scholar]
  27. Vos P., Hogers R., Bleeker M., Reijans M., van de Lee T., Hornes M., Frijters A., Pot J., Peleman J., Kuiper M. AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res. 1995 Nov 11;23(21):4407–4414. doi: 10.1093/nar/23.21.4407. [DOI] [PMC free article] [PubMed] [Google Scholar]

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