1887

Abstract

isolates from cattle and sheep, including representatives of different serotypes and untypable strains, were examined for leukotoxin (Lkt) production at the end of the log phase of growth in brain heart infusion broth. There were marked differences in leukotoxic activity in culture supernate samples, as measured by chemiluminescence-inhibition assays with bovine and ovine neutrophils, even between strains of the same serotype. There was also some variation in the amount and mol. wt of the Lkt protein produced by different strains, as judged by SDS-PAGE, immunoblotting and ELISA. Some strains produced normal amounts of Lkt protein which had only low leukotoxic activity. Most strains produced Lkt of 105 kDa whereas four strains produced a higher mol. wt form of 108 kDa, including two of the five serotype A2 strains examined. Thus, the isolates showed considerable heterogeneity in terms of leukotoxin production, mol. wt and activity, even within a given serotype.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/00222615-46-4-276
1997-04-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jmm/46/4/medmicro-46-4-276.html?itemId=/content/journal/jmm/10.1099/00222615-46-4-276&mimeType=html&fmt=ahah

References

  1. Frank G. H. Pasteurellosis of cattle. In Adlam C., Rutter J. M. (eds) Pasteurella and pasteurellosis London: Academic Press; 1989197–222
    [Google Scholar]
  2. Gilmour N. J.L., Gilmour J. S. Pasteurellosis of sheep. In Adlam C., Rutter J. M. (eds) Pasteurella and pasteurellosis London: Academic Press; 1989223–262
    [Google Scholar]
  3. Sneath P. H., Stevens M. Actinobacillus rossii sp. nov., Actinobacillus seminis sp. nov., nom. rev., Pasteurella bettii sp. nov., Pasteurella lymphangitidis sp. nov., Pasteurella mairi sp. nov., and Pasteurella trehalosi sp. nov. Int J Syst Bacteriol 1990; 40:148–153
    [Google Scholar]
  4. Confer A. W. Immunogens of Pasteurella . Vet Microbiol 1993; 37:353–368
    [Google Scholar]
  5. Conlon J. A., Shewen P. E., Lo R. Y.C. Efficacy of recombinant leukotoxin in protection against pneumonia challenge with live Pasteurella haemolytica Al. Infect Immun 1991; 59:587–591
    [Google Scholar]
  6. Coote J. G. The RTX toxins of Gram-negative bacterial pathogens: modulators of the host immune system. Rev Med Microbiol 1996; 7:53–62
    [Google Scholar]
  7. Chang Y-F, Young R., Post D., Struck D. K. Identification and characterization of the Pasteurella haemolytica leukotoxin. Infect Immun 1987; 55:2348–2354
    [Google Scholar]
  8. Lo R. Y.C., Strathdee C. A., Shewen P. E. Nucleotide sequence of the leukotoxin genes of Pasteurella haemolytica Al. Infect Immun 1987; 55:1987–1996
    [Google Scholar]
  9. Shewen P. E., Wilkie B. N. Cytotoxin of Pasteurella haemolytica acting on bovine leukocytes. Infect Immun 1982; 35:91–94
    [Google Scholar]
  10. Maheswaran S. K., Kannan M. S., Weiss D. J. Enhancement of neutrophil-mediated injury to bovine pulmonary endothelial cells by Pasteurella haemolytica leukotoxin. Infect Immun 1993; 61:2618–2625
    [Google Scholar]
  11. Chang Y. F., Renshaw H. W. Pasteurella haemolytica leukotoxin: comparison of31 chromium-release, trypan blue dye exclusion, and luminol-dependent chemiluminescence-inhibition assays for sensitivity in detecting leukotoxin activity. Am J Vet Res 1986; 47:134–138
    [Google Scholar]
  12. Czuprynski C. J., Noel E. J. Influence of Pasteurella haemolytica Al crude leukotoxin on bovine neutrophil chemiluminescence. Infect Immun 1990; 58:1485–1487
    [Google Scholar]
  13. Ali Q., Davies R. L., Parton R., Coote J. G., Gibbs H. A. Lipopolysaccharide heterogeneity in Pasteurella haemolytica isolates from cattle and sheep. J Gen Microbiol 1992; 138:2185–2195
    [Google Scholar]
  14. Ali Q. Characterisation of virulence-related properties of Pasteurella haemolytica isolates. PhD Thesis 1992 Glasgow University;
    [Google Scholar]
  15. McCluskey J., Gibbs H. A., Davies R. L. Variation in outer-membrane protein and lipopolysaccharide profiles of Pasteurella haemolytica isolates of serotypes Al and A2 obtained from pneumonic and healthy cattle. Microbiology 1994; 140:807–814
    [Google Scholar]
  16. Azad A. K., Coote J. G., Parton R. Distinct plasmid profiles of Pasteurella haemolytica serotypes and the characterization and amplification in Escherichia coli of ampicillin-resistance plasmids encoding ROB-1 β-lactamase. J Gen Microbiol 1992; 138:1185–1196
    [Google Scholar]
  17. Davies R. L., Ali Q., Parton R., Coote J. G., Gibbs H. A., Freer J. H. Optimal conditions for the analysis of Pasteurella haemolytica lipopolysaccharide by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. FEMS Microbiol Lett 1991; 90:23–28
    [Google Scholar]
  18. Davies R. L., Parton R., Coote J. G., Gibbs H. A., Freer J. H. Outer-membrane protein and lipopolysaccharide variation in Pasteurella haemolytica serotype Al under different growth conditions. J Gen Microbiol 1992; 138:909–922
    [Google Scholar]
  19. Shreeve B. J., Biberstein E. L., Thompson D. A. Variation in carrier rates of Pasteurella haemolytica in sheep. II. Diseased flocks. J Comp Pathol 1972; 82:111–118
    [Google Scholar]
  20. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680–685
    [Google Scholar]
  21. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76:4350–4354
    [Google Scholar]
  22. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein, utilizing the principle of protein-dye binding. Anal Biochem 1976; 72:248–257
    [Google Scholar]
  23. Strathdee C. A., Lo R. Y.C. Regulation of expression of the Pasteurella haemolytica leukotoxin determinant. J Bacteriol 1989; 171:5955–5962
    [Google Scholar]
  24. Gatewood D. M., Fenwick B. W., Chengappa M. M. Growth-condition dependent expression of Pasteurella haemolytica Al outer membrane proteins, capsule, and leukotoxin. Vet Microbiol 1994; 41:221–233
    [Google Scholar]
  25. Shewen P. E., Wilkie B. N. Evidence for the Pasteurella haemolytica cytotoxin as a product of actively growing bacteria. Am J Vet Res 1985; 46:1212–1214
    [Google Scholar]
  26. Confer A. W., Durham J. A. Sequential development of antigens and toxins of Pasteurella haemolytica serotype 1 grown in cell culture medium. Am J Vet Res 1992; 53:646–652
    [Google Scholar]
  27. Shewen P. E., Wilkie B. N. Pasteurella haemolytica cytotoxin: production by recognized serotypes and neutralization by type-specific rabbit antisera. Am J Vet Res 1983; 44:715–718
    [Google Scholar]
  28. Chang Y. F., Renshaw H. W., Young R. Pneumonic pasteurellosis: examination of typable and untypable Pasteurella haemolytica strains for leukotoxin production, plasmid content, and antimicrobial susceptibility. Am J Vet Res 1987; 48:378–384
    [Google Scholar]
  29. Gentry M. J., Confer A. W., Holland S. G. Comparison of the toxic and antigenic properties of single bovine isolates of Pasteurella haemolytica representing five serotypes and an untypable strain. Vet Microbiol 1988; 16:351–367
    [Google Scholar]
  30. Gerbig D. G., Cameron M. R., Struck D. K., Moore R. N. Characterization of a neutralizing monoclonal antibody to Pasteurella haemolytica leukotoxin. Infect Immun 1992; 60:1734–1739
    [Google Scholar]
  31. Winfield E. O., Lo R. Y.C. Analysis of the Pasteurella haemolytica T3 leukotoxin determinant. Abstracts of the General Meeting of the American Society for Microbiology; Dallas, Texas: 199133
    [Google Scholar]
  32. Welch R. A. Holistic perspective on the Escherichia coli hemolysin. In Miller V. L., Kaper J. B., Portnoy D. A., Isberg R. R. (eds) Molecular genetics of bacterial pathogenesis. A tribute to Stanley Falkow Washington DC: ASM Press; 1994351–364
    [Google Scholar]
  33. Stanley P. L.D., Diaz P., Bailey M. J.A., Gygi D., Juarez A., Hughes C. Loss of activity in the secreted form of Escherichia coli haemolysin caused by an rfaP lesion in core lipopolysaccharide assembly. Mol Microbiol 1993; 10:781–787
    [Google Scholar]
  34. Lo R. Y.C. Molecular characterisation of the leukotoxin determinants of Pasteurella haemolytica serotypes 1-12. Annual Meeting of The American Society lor Microbiology; Miami, Florida: 1988 Abstract B-60
    [Google Scholar]
  35. Burrows L. L., Olah-Winfield E., Lo R. Y.C. Molecular analysis of the leukotoxin determinants from Pasteurella haemolytica serotypes 1 to 16. Infect Immun 1993; 61:5001–5007
    [Google Scholar]
  36. Frey J., Bosse J. T., Chang Y.-F. Actinobacillus pleuropneumoniae RTX toxins: uniform designation of haemolysins, cytolysins, pleurotoxin and their genes. J Gen Microbiol 1993; 139:1723–1728
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/00222615-46-4-276
Loading
/content/journal/jmm/10.1099/00222615-46-4-276
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