1887

Abstract

was shown to possess peritrichous fimbriae. Electron microscopy of strains FMV 84–01 and ATCC 33209 revealed short, flexible fimbriae less than 2 nm in diameter. These surface appendages were isolated from the bacteria by a procedure involving water extraction and urea solubilization. The fimbrin was purified to homogeneity by Fast Pressure Liquid Chromatography, and shown by SDS-PAGE to be a protein of 57 kDa. Isoelectric focusing under non-denaturing conditions indicated a pI of 4·8. The protein had an amino acid composition rich in glycine, Asx (aspartic acid and asparagine), valine and alanine; methionine was absent. Approximately 33% of the amino acid residues were hydrophobic. Immunoblotting using a polyclonal antiserum raised against whole cells showed that the 57 kDa protein was the immunodominant antigen on the cell surface. Immunogold labelling using polyclonal antibodies raised against the fimbrin revealed an alignment of gold particles along the fimbriae. Purified fimbriae caused agglutination of rabbit erythrocytes and antifimbrial serum inhibited this haemagglutination. Altogether the results indicate that the fimbriae on the surface of are responsible for the haemagglutinating activity.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-136-12-2443
1990-12-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/136/12/mic-136-12-2443.html?itemId=/content/journal/micro/10.1099/00221287-136-12-2443&mimeType=html&fmt=ahah

References

  1. Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. 1984; A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on western blots. Analytical Biochemistry 136:175–179
    [Google Scholar]
  2. Bruno D. W. 1986; Histopathology of bacterial kidney disease in laboratory infected rainbow trout, Salmo gairdneri Richardson, and Atlantic salmon, Salmo salar L., with reference to naturally infected fish. Journal of Fish Diseases 9:523–537
    [Google Scholar]
  3. Bruno D. W. 1988; The relationship between auto-agglutination, cell surface hydrophobicity and virulence of the fish pathogen Renibacterium salmoninarum . FEMS Microbiology Letters 51:135–140
    [Google Scholar]
  4. Bruno D. W. 1990; Presence of a saline extractable protein associated with virulent strains of the fish pathogen, Renibacterium salmoninarum . Bulletin of the European Association of Fish Pathologists 10:8–10
    [Google Scholar]
  5. Bruno D. W., Munro A. L. S. 1986; Uniformity in the biochemical properties of Renibacterium salmoninarum isolates obtained from several sources. FEMS Microbiology Letters 33:247–250
    [Google Scholar]
  6. Bullock G. L., Sutckey H. M., Chen P. K. 1974; Corynebacterial kidney disease of salmonids: growth and serological studies on the causative bacterium. Applied Microbiology 28:811–814
    [Google Scholar]
  7. Daly J. G., Stevenson R. M. W. 1985; Charcoal agar, a new growth medium for the fish disease bacterium Renibacterium salmoninarum . Applied and Environmental Microbiology 50:868–871
    [Google Scholar]
  8. Daly J. G., Stevenson R. M. W. 1987; Hydrophobic and haemagglutinating properties of Renibacterium salmoninarum . Journal of General Microbiology 133:3575–3580
    [Google Scholar]
  9. Daly J. G., Stevenson R. M. W. 1989; Agglutination of salmonid spermatozoa by Renibacterium salmoninarum . Journal of Aquatic Animal Health 1:163–164
    [Google Scholar]
  10. Daly J. G., Stevenson R. M. W. 1990; Characterization of the Renibacterium salmoninarum haemagglutinin. Journal of General Microbiology 136:949–953
    [Google Scholar]
  11. Dubreuil D., Lallier R., Jacques M. 1990; Immunoelectron microscopic demonstration that Renibacterium salmoninarum is encapsulated. FEMS Microbiology Letters 66:313–316
    [Google Scholar]
  12. Fairbanks G., Steck T. L., Wallach D. F. H. 1971; Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10:2606–2617
    [Google Scholar]
  13. Fiedler F., Draxl R. 1986; Biochemical and immunochemical properties of the cell surface of Renibacterium salmoninarum . Journal of Bacteriology 168:799–804
    [Google Scholar]
  14. Fortin M., Jacques M. 1987; Hemagglutination by Pasteurella multocida of porcine origin. Journal of Clinical Microbiology 25:938–939
    [Google Scholar]
  15. Fryer J. L., Sanders J. E. 1981; Bacterial kidney disease of salmonid fish. Annual Review of Microbiology 35:273–298
    [Google Scholar]
  16. Getchell R. G., Rohovec J. S., Fryer J. L. 1985; Comparison of Renibacterium salmoninarum isolates by antigenic analysis. Fish Pathology 20:149–159
    [Google Scholar]
  17. Gubish E. R., Chen K. D. S., Buchanan T. M. 1982; Attachment of gonococcal lectin-resistant clones of Chinese hamster ovary cells. Infection and Immunity 37P:189–194
    [Google Scholar]
  18. Isaacson R. E. 1985; Pilus adhesins. In Bacterial Adhesion pp. 307–336 Savage D. C., Fletcher M. Edited by New York: Plenum;
    [Google Scholar]
  19. Isaacson R. E., Richter P. 1981; Escherichia coli pilus: purification and partial characterisation. Journal of Bacteriology 146:784–789
    [Google Scholar]
  20. Kusser W., Fiedler F. 1983; Murein type and polysaccharide composition of cell walls from Renibacterium salmoninarum . FEMS Microbiology Letters 20:391–394
    [Google Scholar]
  21. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  22. Legendre N., Matsudaira P. 1988; Direct protein microsequencing from Immobilon-P transfer membrane. Biotechniques 6:154–159
    [Google Scholar]
  23. Masuda N., Ellen R. P., Grove D. A. 1981; Purification and characterisation of surface fibrils from taxonomically typical Actinomyces viscosus WYU627. Journal of Bacteriology 147:1095–1104
    [Google Scholar]
  24. Ottow J. C. G. 1975; Ecology, physiology and genetics of fimbriae and pili. Annual Review of Microbiology 29:79–108
    [Google Scholar]
  25. Robertson E. F., Dannelly H. K., Malloy P. J., Reeves H. C. 1987; Rapid isoelectrofocusing in a vertical polyacrylamide minigel system. Analytical Biochemistry 167:290–294
    [Google Scholar]
  26. Salinovich O., Montelaro R. 1986; Reversible staining and peptide mapping of proteins transferred to nitrocellulose after separation by sodium dodecylsulfate polyacrylamide gel electrophoresis. Analytical Biochemistry 156:341–347
    [Google Scholar]
  27. Sanders J. E., Fryer J. L. 1986; Genus Renibacterium Sanders and Fryer 1980, 501. In Bergey’s Manual of Systematic Bacteriology 2 pp. 1253–1254 Sneath P. H. A., Mair N. S., Sharpe M. E., Holt J. G. Edited by Baltimore: Williams & Wilkins;
    [Google Scholar]
  28. Smith I. W. 1964; The occurrence and pathology of Dee disease. Freshwater Salmon Research 34:1–12
    [Google Scholar]
  29. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America 76:4350–4354
    [Google Scholar]
  30. Turaga P. S. D., Weins G. D., Kaattari S. L. 1987; Analysis of Renibacterium salmoninarum antigen production in situ . Fish Pathology 22:209–214
    [Google Scholar]
  31. Young C. L., Chapman G. B. 1978; Ultrastructural aspects of the causative agent and renal histopathology of bacterial kidney disease in brook trout (Salvelinus fontinalis). Journal of the Fisheries Research Board of Canada 35:1234–1248
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-136-12-2443
Loading
/content/journal/micro/10.1099/00221287-136-12-2443
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error