Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1996 Mar;62(3):918–927. doi: 10.1128/aem.62.3.918-927.1996

Phenotypic characterization of Vibrio vulnificus biotype 2, a lipopolysaccharide-based homogeneous O serogroup within Vibrio vulnificus.

E G Biosca 1, J D Oliver 1, C Amaro 1
PMCID: PMC167856  PMID: 8975619

Abstract

In this study, we have reevaluated the taxonomic position of biotype 2 of Vibrio vulnificus. For this purpose, we have biochemically and serologically characterized 83 biotype 2 strains from diseased eels, comparing them with 17 biotype 1 strains from different sources. Selected strains were also molecularly analyzed and tested for eel and mouse pathogenicity. Results have shown that biotype 2 (i) is biochemically homogeneous, indole production being the main trait that distinguishes it from biotype 1, (ii) presents small variations in DNA restriction profiles and outer membrane protein patterns, some proteins being immunologically related to outer membrane proteins from biotype 1, (iii) expresses a common lipopolysaccharide (LPS) profile, which is immunologically identical among strains and distinct from that of LPS of tested biotype 1 strains, and (iv) contains at least two high-Mr plasmids. Regarding host range, we have confirmed that both biotypes are pathogenic for mice but only biotype 2 is pathogenic for eels. On the basis of these data, we propose that biotype 2 of V. vulnificus constitutes an LPS-based O serogroup which is phenotypically homogeneous and pathogenic for eels. In this article, the serogroup is designated serogroup E (for eels).

Full Text

The Full Text of this article is available as a PDF (581.0 KB).

Selected References

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

  1. Amaro C., Aznar R., Garay E., Alcaide E. R plasmids in environmental Vibrio cholerae non-O1 strains. Appl Environ Microbiol. 1988 Nov;54(11):2771–2776. doi: 10.1128/aem.54.11.2771-2776.1988. [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. Bernoth E. M. Marketing authorisation of fish drugs--current status and future intentions. Zentralbl Veterinarmed B. 1990 Aug;37(6):401–406. doi: 10.1111/j.1439-0450.1990.tb01075.x. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Biosca E. G., Llorens H., Garay E., Amaro C. Presence of a capsule in Vibrio vulnificus biotype 2 and its relationship to virulence for eels. Infect Immun. 1993 May;61(5):1611–1618. doi: 10.1128/iai.61.5.1611-1618.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bowdre J. H., Hull J. H., Cocchetto D. M. Antibiotic efficacy against Vibrio vulnificus in the mouse: superiority of tetracycline. J Pharmacol Exp Ther. 1983 Jun;225(3):595–598. [PubMed] [Google Scholar]
  8. Buck J. D. Nonstaining (KOH) method for determination of gram reactions of marine bacteria. Appl Environ Microbiol. 1982 Oct;44(4):992–993. doi: 10.1128/aem.44.4.992-993.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cipriano R. C., Pyle J. B., Starliper C. E., Pyle S. W. Detection of Vibrio anguillarum antigen by the dot blot assay. J Wildl Dis. 1985 Jul;21(3):211–218. doi: 10.7589/0090-3558-21.3.211. [DOI] [PubMed] [Google Scholar]
  10. Cullmann W., Stieglitz M., Baars B., Opferkuch W. Comparative evaluation of recently developed quinolone compounds--with a note on the frequency of resistant mutants. Chemotherapy. 1985;31(1):19–28. doi: 10.1159/000238309. [DOI] [PubMed] [Google Scholar]
  11. Davidson L. S., Oliver J. D. Plasmid carriage in Vibrio vulnificus and other lactose-fermenting marine vibrios. Appl Environ Microbiol. 1986 Jul;52(1):211–213. doi: 10.1128/aem.52.1.211-213.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Heimbrook M. E., Wang W. L., Campbell G. Staining bacterial flagella easily. J Clin Microbiol. 1989 Nov;27(11):2612–2615. doi: 10.1128/jcm.27.11.2612-2615.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Janda J. M., Bottone E. J. Pseudomonas aeruginosa enzyme profiling: predictor of potential invasiveness and use as an epidemiological tool. J Clin Microbiol. 1981 Jul;14(1):55–60. doi: 10.1128/jcm.14.1.55-60.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kitamikado M., Lee Y. Z. Chondroitinase-producing bacteria in natural habitats. Appl Microbiol. 1975 Mar;29(3):414–421. doi: 10.1128/am.29.3.414-421.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lee J. V., Bashford D. J., Donovan T. J., Furniss A. L., West P. A. The incidence of Vibrio cholerae in water, animals and birds in Kent, England. J Appl Bacteriol. 1982 Apr;52(2):281–291. doi: 10.1111/j.1365-2672.1982.tb04852.x. [DOI] [PubMed] [Google Scholar]
  18. Martin S. J., Siebeling R. J. Identification of Vibrio vulnificus O serovars with antilipopolysaccharide monoclonal antibody. J Clin Microbiol. 1991 Aug;29(8):1684–1688. doi: 10.1128/jcm.29.8.1684-1688.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Massad G., Oliver J. D. New selective and differential medium for Vibrio cholerae and Vibrio vulnificus. Appl Environ Microbiol. 1987 Sep;53(9):2262–2264. doi: 10.1128/aem.53.9.2262-2264.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Morris J. G., Jr, Tenney J. Antibiotic therapy for Vibrio vulnificus infection. JAMA. 1985 Feb 22;253(8):1121–1122. [PubMed] [Google Scholar]
  21. Oliver J. D., Guthrie K., Preyer J., Wright A., Simpson L. M., Siebeling R., Morris J. G., Jr Use of colistin-polymyxin B-cellobiose agar for isolation of Vibrio vulnificus from the environment. Appl Environ Microbiol. 1992 Feb;58(2):737–739. doi: 10.1128/aem.58.2.737-739.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Oliver J. D., Wear J. E., Thomas M. B., Warner M., Linder K. Production of extracellular enzymes and cytotoxicity by Vibrio vulnificus. Diagn Microbiol Infect Dis. 1986 Jul;5(2):99–111. doi: 10.1016/0732-8893(86)90112-4. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Schwyn B., Neilands J. B. Universal chemical assay for the detection and determination of siderophores. Anal Biochem. 1987 Jan;160(1):47–56. doi: 10.1016/0003-2697(87)90612-9. [DOI] [PubMed] [Google Scholar]
  25. Shimada T., Sakazaki R. On the serology of Vibrio vulnificus. Jpn J Med Sci Biol. 1984 Oct-Dec;37(5-6):241–246. doi: 10.7883/yoken1952.37.241. [DOI] [PubMed] [Google Scholar]
  26. Simpson L. M., Oliver J. D. Siderophore production by Vibrio vulnificus. Infect Immun. 1983 Aug;41(2):644–649. doi: 10.1128/iai.41.2.644-649.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stelma G. N., Jr, Reyes A. L., Peeler J. T., Johnson C. H., Spaulding P. L. Virulence characteristics of clinical and environmental isolates of Vibrio vulnificus. Appl Environ Microbiol. 1992 Sep;58(9):2776–2782. doi: 10.1128/aem.58.9.2776-2782.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Szu S. C., Schneerson R., Robbins J. B. Rabbit antibodies to the cell wall polysaccharide of Streptococcus pneumoniae fail to protect mice from lethal challenge with encapsulated pneumococci. Infect Immun. 1986 Nov;54(2):448–455. doi: 10.1128/iai.54.2.448-455.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Venkateswaran K., Nakano H., Okabe T., Takayama K., Matsuda O., Hashimoto H. Occurrence and distribution of Vibrio spp., Listonella spp., and Clostridium botulinum in the Seto Inland Sea of Japan. Appl Environ Microbiol. 1989 Mar;55(3):559–567. doi: 10.1128/aem.55.3.559-567.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wright A. C., Miceli G. A., Landry W. L., Christy J. B., Watkins W. D., Morris J. G., Jr Rapid identification of Vibrio vulnificus on nonselective media with an alkaline phosphatase-labeled oligonucleotide probe. Appl Environ Microbiol. 1993 Feb;59(2):541–546. doi: 10.1128/aem.59.2.541-546.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES