Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1997 Apr;63(4):1449–1452. doi: 10.1128/aem.63.4.1449-1452.1997

Intraspecific characterization of Vibrio tapetis strains by use of pulsed-field gel electrophoresis, ribotyping, and plasmid profiling.

D Castro 1, J L Romalde 1, J Vila 1, B Magariños 1, A Luque 1, J J Borrego 1
PMCID: PMC168440  PMID: 9097443

Abstract

A total of twenty-two strains of Vibrio tapetis, the causative agent of brown ring disease affecting cultured clams, were compared and evaluated in an investigation of strain heterogeneity using pulsed-field gel electrophoresis (PFGE), ribotyping, and plasmid profile analysis. A total of 90.9% of V. tapetis strains tested by using NotI showed the same PFGE pattern, consisting of 15 bands. In contrast, the V. tapetis strains showed a low degree of similarity with six reference Vibrio species tested. All V. tapetis strains harbored a large plasmid of 74.5 kb. This plasmid was not detected in any of the other Vibrio species. In addition, endonuclease restriction analysis of the plasmid content of the strains using EcoRI and HindIII clearly showed that all the strains of V. tapetis possessed the same cleavage pattern. The three enzymes used for ribotyping, PvuII, SmaI, and SalI, yielded patterns with 8 to 12 bands ranging in size from 2 to 23 kb. The application of the SalI and SmaI endonuclease rendered the separation of the strains tested in two ribotypes, while all the V. tapetis strains belonged to the same ribotype when the enzyme PvuII was used.

Full Text

The Full Text of this article is available as a PDF (566.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bingen E. H., Denamur E., Elion J. Use of ribotyping in epidemiological surveillance of nosocomial outbreaks. Clin Microbiol Rev. 1994 Jul;7(3):311–327. doi: 10.1128/cmr.7.3.311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Borrego J. J., Moriñigo M. A., Martinez-Manzanares E., Bosca M., Castro D., Barja J. L., Toranzo A. E. Plasmid associated virulence properties of environmental isolates of Aeromonas hydrophila. J Med Microbiol. 1991 Nov;35(5):264–269. doi: 10.1099/00222615-35-5-264. [DOI] [PubMed] [Google Scholar]
  4. Chachaty E., Saulnier P., Martin A., Mario N., Andremont A. Comparison of ribotyping, pulsed-field gel electrophoresis and random amplified polymorphic DNA for typing Clostridium difficile strains. FEMS Microbiol Lett. 1994 Sep 15;122(1-2):61–68. doi: 10.1111/j.1574-6968.1994.tb07144.x. [DOI] [PubMed] [Google Scholar]
  5. Crosa J. H. A plasmid associated with virulence in the marine fish pathogen Vibrio anguillarum specifies an iron-sequestering system. Nature. 1980 Apr 10;284(5756):566–568. doi: 10.1038/284566a0. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Gordillo M. E., Singh K. V., Murray B. E. Comparison of ribotyping and pulsed-field gel electrophoresis for subspecies differentiation of strains of Enterococcus faecalis. J Clin Microbiol. 1993 Jun;31(6):1570–1574. doi: 10.1128/jcm.31.6.1570-1574.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grimont F., Grimont P. A. Ribosomal ribonucleic acid gene restriction patterns as potential taxonomic tools. Ann Inst Pasteur Microbiol. 1986 Sep-Oct;137B(2):165–175. doi: 10.1016/s0769-2609(86)80105-3. [DOI] [PubMed] [Google Scholar]
  9. Haertl R., Bandlow G. Epidemiological fingerprinting of Enterobacter cloacae by small-fragment restriction endonuclease analysis and pulsed-field gel electrophoresis of genomic restriction fragments. J Clin Microbiol. 1993 Jan;31(1):128–133. doi: 10.1128/jcm.31.1.128-133.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Herrmann J. L., Bellenger E., Perolat P., Baranton G., Saint Girons I. Pulsed-field gel electrophoresis of NotI digests of leptospiral DNA: a new rapid method of serovar identification. J Clin Microbiol. 1992 Jul;30(7):1696–1702. doi: 10.1128/jcm.30.7.1696-1702.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Larsen J. L., Olsen J. E. Occurrence of plasmids in Danish isolates of Vibrio anguillarum serovars O1 and O2 and association of plasmids with phenotypic characteristics. Appl Environ Microbiol. 1991 Aug;57(8):2158–2163. doi: 10.1128/aem.57.8.2158-2163.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Olsen J. E., Larsen J. L. Restriction fragment length polymorphism of the Vibrio anguillarum serovar O1 virulence plasmid. Appl Environ Microbiol. 1990 Oct;56(10):3130–3132. doi: 10.1128/aem.56.10.3130-3132.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Olsen J. E., Skov M. N., Threlfall E. J., Brown D. J. Clonal lines of Salmonella enterica serotype Enteritidis documented by IS200-, ribo-, pulsed-field gel electrophoresis and RFLP typing. J Med Microbiol. 1994 Jan;40(1):15–22. doi: 10.1099/00222615-40-1-15. [DOI] [PubMed] [Google Scholar]
  15. Olsen J. E., Skov M. Genomic lineage of Salmonella enterica serovar Dublin. Vet Microbiol. 1994 Jun;40(3-4):271–282. doi: 10.1016/0378-1135(94)90116-3. [DOI] [PubMed] [Google Scholar]
  16. Platt D. J., Taggart J., Heraghty K. A. Molecular divergence of the serotype-specific plasmid (pSLT) among strains of Salmonella typhimurium of human and veterinary origin and comparison of pSLT with the serotype specific plasmids of S. enteritidis and S. dublin. J Med Microbiol. 1988 Dec;27(4):277–284. doi: 10.1099/00222615-27-4-277. [DOI] [PubMed] [Google Scholar]
  17. Prevost G., Jaulhac B., Piemont Y. DNA fingerprinting by pulsed-field gel electrophoresis is more effective than ribotyping in distinguishing among methicillin-resistant Staphylococcus aureus isolates. J Clin Microbiol. 1992 Apr;30(4):967–973. doi: 10.1128/jcm.30.4.967-973.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Tiainen T., Pedersen K., Larsen J. L. Ribotyping and plasmid profiling of Vibrio anguillarum serovar O2 and Vibrio ordalii. J Appl Bacteriol. 1995 Oct;79(4):384–392. doi: 10.1111/j.1365-2672.1995.tb03152.x. [DOI] [PubMed] [Google Scholar]
  20. Tolmasky M. E., Actis L. A., Toranzo A. E., Barja J. L., Crosa J. H. Plasmids mediating iron uptake in Vibrio anguillarum strains isolated from turbot in Spain. J Gen Microbiol. 1985 Aug;131(8):1989–1997. doi: 10.1099/00221287-131-8-1989. [DOI] [PubMed] [Google Scholar]
  21. Toranzo A. E., Barja J. L., Colwell R. R., Hetrick F. M. Characterization of plasmids in bacterial fish pathogen. Infect Immun. 1983 Jan;39(1):184–192. doi: 10.1128/iai.39.1.184-192.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES