1887

Abstract

The outer-membrane protein (OMP) and lipopolysaccharide (LPS) profiles of 29 isolates of serotypes A1 (18 isolates) and A2 (11 isolates), obtained from pneumonic (13 isolates) or healthy (16 isolates) cattle, were compared by SDS-PAGE and Western blot analysis. Coomassie-blue-stained OMP profiles of serotype A1 and A2 isolates could be distinguished from each other by differences in both major and minor proteins. Whereas the OMP profiles of the serotype A1 isolates were extremely uniform in stained gels, there was variation in the mobilities of high-molecular-mass minor proteins and one of the major proteins of serotype A2 isolates. Differences in the OMP profiles of isolates within both the A1 and A2 serotypes were more clearly distinguished by Western blotting than by staining after SDS-PAGE. Thus, by Western blot analysis, four distinct OMP profiles were identified within the serotype A1 and A2 isolates, respectively. The profiles of the serotype A1 isolates were designated OMP types 1.1, 1.2, 1.3 and 1.4; those of the serotype A2 isolates were designated OMP types 2.1, 2.2, 2.3 and 2.4. Three distinct LPS profiles were recognized among the isolates which, by comparison with previously described LPS types, were identified as smooth LPS type 1 and rough LPS types 3 and 5. Isolates of serotype A1 consisted of LPS type 1 only, whereas isolates of serotype A2 consisted of LPS types 3 or 5. OMP and LPS analysis of has applications in epidemiological and virulence studies.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-140-4-807
1994-04-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/140/4/mic-140-4-807.html?itemId=/content/journal/micro/10.1099/00221287-140-4-807&mimeType=html&fmt=ahah

References

  1. Achtman M., Mercer A., Kusecek B., Heusenroeder M., Aaronson W.F., Sutton A., Silver R.P. Six widespread bacterial clones among’Escherichia coli K1 isolates. Infect Immun 1983; 39:315–335
    [Google Scholar]
  2. Achtman M., Heusenroeder M., Kusecek B., Ochman H., Caugant D., Selander R.K., Vaisanen-Rhen V., Stuart S., Orskov F., Orskov I. Clonal analysis of Escherichia coli 02: K1 isolated from diseased humans and animals. Infect Immun 1986; 51:268–276
    [Google Scholar]
  3. Adlam C., Knights J.M., Mugridge A., Lindon J.C., Baker P.R.W., Beesley J.E., Spacey B., Craig G.R., Nagy L.K. Purification, characterization and immunological properties of the serotype-specific capsular polysaccharide of Pasteurella haemolytica (serotype Al) organisms. J Gen Microbiol 1984; 130:2415–2426
    [Google Scholar]
  4. Adlam C., Knights J.M., Mugridge A., Williams I.M., Lindon J.C. Production of colominic acid by Pasteurella haemolytica serotype A2 organisms. FEMS Microbiol Lett 1987; 42:23–25
    [Google Scholar]
  5. Ali Q., Davies R.L., Parton R., Coote J.G., Gibbs H.A. Lipopolysaccharide heterogeneity in Pasteurella haemolytica isolates from cattle amd sheep. J Gen Microbiol 1992; 138:2185–2195
    [Google Scholar]
  6. Biberstein E.L. Biotyping and serotyping of Pasteurella haemolytica. Methods Microbiol 1978; 10:253–269
    [Google Scholar]
  7. Blaser M.J., Hopkins A.J., Berka R.M., Vasil M.L., Quang W.L.L. Identification and characterization of Campylobacter jejuni outer membrane proteins. Inject Immun 1983; 42:276–284
    [Google Scholar]
  8. Bolstad A.I., Kristoffersen T., Olsen I., Preus H.R., Jensen H.B., Vasstrand E.N., Bakken V. Outer membrane proteins of Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus studied by SDS-PAGE and immunoblotting. Oral Microbiol Immunol 1990; 5:155–161
    [Google Scholar]
  9. Davies R.L. Outer membrane protein profiles of Yersina ruckeri. Vet Microbiol 1991; 26:125–140
    [Google Scholar]
  10. Davies R.L., Wall R.A., Borriello S.P. Comparison of methods for the analysis of outer membrane antigens of Neisseria meningitidis. J Immunol Methods 1990; 134:215–225
    [Google Scholar]
  11. 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 Eett 1991; 90:3–28
    [Google Scholar]
  12. Davies R.L., Parton R., Coote J.G., Gibbs H.A., Freer J.H. Outer-membrane protein and lipopolysaccharide variation in Pasteurella haemolytica A1 under different growth conditions. J Gen Microbiol 1992; 138:909–922
    [Google Scholar]
  13. Deneer H.G., Potter A.A. Iron-repressible outer membrane proteins of Pasteurella haemolytica. J Gen Microbiol 1989; 135:435–443
    [Google Scholar]
  14. Filip G., Fletcher G., Wulff J.L., Earhart C.F. Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacterial 1973; 115:717–722
    [Google Scholar]
  15. Fodor L., Varga J., Hajtos J., Donachie W., Gilmour N.J.L. New serotypes of Pasteurella haemolytica isolates in Hungary. Vet Res 1988; 121:155
    [Google Scholar]
  16. Frank G.H. Pasteurellosis of cattle. In Pasteurella and Pasteurellosis 1989 Edited by Adlam C.F., Rutter J.M. London: Academic Press; pp 197–222
    [Google Scholar]
  17. Gilmour N.J.L., Gilmour J.S. Pasteurellosis of sheep. In Pasteurella and Pasteurellosis 1989 Edited by Adlam C.F., Rutter J.M. London: Academic Press; pp 223–261
    [Google Scholar]
  18. Hansman D., Lawrence A. Outer membrane protein and immunoblot analysis of Australian isolates of Haemophilus influenzae. J Med Microbiol 1993; 38:378–383
    [Google Scholar]
  19. Hitchcock P.J., Brown T.M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol 1983; 154:269–277
    [Google Scholar]
  20. Inzana T.J. Electrophoretic heterogeneity and interstrain variation of the lipopolysaccharide of Haemophilus influenzae. J Infect Dis 1983; 148:492–499
    [Google Scholar]
  21. Inzana T.J., Pichichero M.E. Lipopolysaccharide subtypes of Haemophilus influenzae type b from an outbreak of invasive disease. J Clin Microbiol 1984; 20:145–150
    [Google Scholar]
  22. Knights J.M., Adlam C., Owen P. Characterization of envelope proteins from Pasteurella haemolytica and Pasteurella multocida. J Gen Microbiol 1990; 136:495–505
    [Google Scholar]
  23. Lacroix R.P., Duncan J.R., Jenkins R.P., Leitch R.A., Perry J.A., Richards J.C. Structural and serological specificities of Pasteurella haemolytica lipopolysaccharides. Infect Immun 1993; 61:170–181
    [Google Scholar]
  24. Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680–685
    [Google Scholar]
  25. Loeb M.R., Smith D.A. Outer membrane protein composition in disease isolates of Haemophilus influenzae: pathogenic and epidemiological implications. Infect Immun 1980; 30:709–717
    [Google Scholar]
  26. Lugtenberg B., Boxtel R., Dejong M. Atrophic rhinitis in swine: correlation of Pasteurella multocida pathogenicity with membrane proteins and lipopolysaccharide patterns. Infect Immun 1984; 46:48–54
    [Google Scholar]
  27. Markwell M.A.K., Haas S.M., Bieber L.L., 8, Tolbert N.E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem 1978; 87:206–210
    [Google Scholar]
  28. Mocca L.F., Frasch C.E. Sodium dodecyl sulphate-polyacrylamide gel typing system for characterization of Neisseria meningitidis. J Clin Microbiol 1982; 16:240–244
    [Google Scholar]
  29. Mulligan M.E., Peterson L.R., Kwok R.Y.Y., Clabots C.R., Gerding D.N. Immunoblots and plasmid fingerprints compared with serotyping and polyacrylamide gel electrophoresis for typing Clostridium difficile. J Clin Microbiol 1988; 26:41–46
    [Google Scholar]
  30. Odumeru J.A., Ronald A.R., Albritton W.L. Characterization of cell proteins of Haemophilus ducreyi by polyacrylamide gel electrophoresis. J Infect Dis 1983; 148:710–714
    [Google Scholar]
  31. Poxton I.R., Bell G.T., Barclay G.R. The association on SDS-polyacrylamide gels of lipopolysaccharide and outer membrane proteins of Pseudomonas aeruginosa as revealed by monoclonal antibodies and Western blotting. FEMS Microbiol Eett 1985; 27:247–251
    [Google Scholar]
  32. Rapp V.J., Munson R.S., Ross R.F. Outer membrane protein profiles of Haemophilus pleuropneumoniae. Infect Immun 1986; 52:414–420
    [Google Scholar]
  33. 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]
  34. 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]
  35. Squire P.G., Smiley D.N., Croskell R.B. Identification and extraction of Pasteurella haemolytica membrane proteins. Infect Immun 1984; 45:667–673
    [Google Scholar]
  36. Tsai C.M., Frasch C.E. A sensitive silver stain for detecting lipopolysaccharide in polyacrylamide gels. Anal Biochem 1982; 119:115–119
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-140-4-807
Loading
/content/journal/micro/10.1099/00221287-140-4-807
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