Abstract
Outer membrane protein profiles of Haemophilus pleuropneumoniae were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cells were disrupted by sonication, and outer membrane-enriched fractions were prepared by differential centrifugation and selective solubilization of the inner membrane with sodium N-lauroyl sarcosinate. Colony type, growth medium, time of harvest, and in vitro or in vivo passage had no appreciable effect on the protein profiles of the strains examined. Seven patterns were distinguished among the reference strains of the nine capsular serotypes. These patterns were based on the mobility of the major outer membrane proteins migrating in the 39,000- to 44,000-molecular-weight region of the gel, a 16K to 16.5K protein, and a heat-modifiable 29K protein. Strains of serotypes 1 and 9 had identical outer membrane protein profiles, as did strains of serotypes 2 and 6. The reference strains of the remaining five serotypes each had a distinct pattern. The outer membrane protein profiles of 95 field isolates belonging to serotypes 1, 5, 7, and 9 from swine in the midwestern United States were determined and compared with the reference patterns. The results indicate that the population of H. pleuropneumoniae is clonal, with three predominant clones distinguished by both serotype and outer membrane protein profile responsible for the majority of H. pleuropneumoniae disease occurring in swine in the United States.
Full text
PDF






Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Achtman M., Mercer A., Kusecek B., Pohl A., Heuzenroeder M., Aaronson W., Sutton A., Silver R. P. Six widespread bacterial clones among Escherichia coli K1 isolates. Infect Immun. 1983 Jan;39(1):315–335. doi: 10.1128/iai.39.1.315-335.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ames G. F. Resolution of bacterial proteins by polyacrylamide gel electrophoresis on slabs. Membrane, soluble, and periplasmic fractions. J Biol Chem. 1974 Jan 25;249(2):634–644. [PubMed] [Google Scholar]
- Barenkamp S. J., Munson R. S., Jr, Granoff D. M. Subtyping isolates of Haemophilus influenzae type b by outer-membrane protein profiles. J Infect Dis. 1981 May;143(5):668–676. doi: 10.1093/infdis/143.5.668. [DOI] [PubMed] [Google Scholar]
- Beher M. G., Schnaitman C. A., Pugsley A. P. Major heat-modifiable outer membrane protein in gram-negative bacteria: comparison with the ompA protein of Escherichia coli. J Bacteriol. 1980 Aug;143(2):906–913. doi: 10.1128/jb.143.2.906-913.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chopra I., Shales S. W. Comparison of the polypeptide composition of Escherichia coli outer membranes prepared by two methods. J Bacteriol. 1980 Oct;144(1):425–427. doi: 10.1128/jb.144.1.425-427.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coulton J. W., Wan D. T. The outer membrane of haemophilus influenzae type b: cell envelope associations of major proteins. Can J Microbiol. 1983 Feb;29(2):280–287. doi: 10.1139/m83-046. [DOI] [PubMed] [Google Scholar]
- Ezzell J. W., Dobrogosz W. J., Kloos W. E., Manclark C. R. Phase-shift markers in Bordetella: alterations in envelope proteins. J Infect Dis. 1981 Apr;143(4):562–569. doi: 10.1093/infdis/143.4.562. [DOI] [PubMed] [Google Scholar]
- Filip C., Fletcher G., Wulff J. L., Earhart C. F. Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol. 1973 Sep;115(3):717–722. doi: 10.1128/jb.115.3.717-722.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gunnarsson A., Biberstein E. L., Hurvell B. Serologic studies on porcine strains of Haemophilus parahaemolyticus (pleuropneumoniae): agglutination reactions. Am J Vet Res. 1977 Aug;38(8):1111–1114. [PubMed] [Google Scholar]
- Hancock R. E., Carey A. M. Outer membrane of Pseudomonas aeruginosa: heat- 2-mercaptoethanol-modifiable proteins. J Bacteriol. 1979 Dec;140(3):902–910. doi: 10.1128/jb.140.3.902-910.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Lambden P. R., Heckels J. E., James L. T., Watt P. J. Variations in surface protein composition associated with virulence properties in opacity types of Neisseria gonorrhoeae. J Gen Microbiol. 1979 Oct;114(2):305–312. doi: 10.1099/00221287-114-2-305. [DOI] [PubMed] [Google Scholar]
- Loeb M. R., Smith D. H. Outer membrane protein composition in disease isolates of Haemophilus influenzae: pathogenic and epidemiological implications. Infect Immun. 1980 Dec;30(3):709–717. doi: 10.1128/iai.30.3.709-717.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Martin P. G., Lachance P., Niven D. F. Production of RNA-dependent haemolysin by Haemophilus pleuropneumoniae. Can J Microbiol. 1985 May;31(5):456–462. doi: 10.1139/m85-085. [DOI] [PubMed] [Google Scholar]
- Murphy T. F., Dudas K. C., Mylotte J. M., Apicella M. A. A subtyping system for nontypable Haemophilus influenzae based on outer-membrane proteins. J Infect Dis. 1983 May;147(5):838–846. doi: 10.1093/infdis/147.5.838. [DOI] [PubMed] [Google Scholar]
- Musser J. M., Granoff D. M., Pattison P. E., Selander R. K. A population genetic framework for the study of invasive diseases caused by serotype b strains of Haemophilus influenzae. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5078–5082. doi: 10.1073/pnas.82.15.5078. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nicolet J. Sur l'hémophilose du pore. 3. Différenciation sérologique de Haemophilus parahaemolyticus. Zentralbl Bakteriol Orig. 1971;216(4):487–495. [PubMed] [Google Scholar]
- Nielsen R., O'Connor P. J. Serological characterization of 8 Haemophilus pleuropneumoniae strains and proposal of a new serotype: serotype 8. Acta Vet Scand. 1984;25(1):96–106. doi: 10.1186/BF03547283. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ochman H., Selander R. K. Evidence for clonal population structure in Escherichia coli. Proc Natl Acad Sci U S A. 1984 Jan;81(1):198–201. doi: 10.1073/pnas.81.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Odumeru J. A., Ronald A. R., Albritton W. L. Characterization of cell proteins of Haemophilus ducreyi by polyacrylamide gel electrophoresis. J Infect Dis. 1983 Oct;148(4):710–714. doi: 10.1093/infdis/148.4.710. [DOI] [PubMed] [Google Scholar]
- Parton R., Wardlaw A. C. Cell-envelope proteins of Bordetella pertussis. J Med Microbiol. 1975 Feb;8(1):47–57. doi: 10.1099/00222615-8-1-47. [DOI] [PubMed] [Google Scholar]
- Pedersen K. B., Froholm L. O., Bovre K. Fimbriation and colony type of Moraxella bovis in relation to conjunctival colonization and development of keratoconjunctivitis in cattle. Acta Pathol Microbiol Scand B Microbiol Immunol. 1972;80(6):911–918. doi: 10.1111/j.0365-5563.1973.tb00019.x. [DOI] [PubMed] [Google Scholar]
- Peppler M. S., Schrumpf M. E. Phenotypic variation and modulation in Bordetella bronchiseptica. Infect Immun. 1984 Jun;44(3):681–687. doi: 10.1128/iai.44.3.681-687.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
- Rapp V. J., Ross R. F., Erickson B. Z. Serotyping of Haemophilus pleuropneumoniae by rapid slide agglutination and indirect fluorescent antibody tests in swine. Am J Vet Res. 1985 Jan;46(1):185–192. [PubMed] [Google Scholar]
- Rosendal S., Boyd D. A. Haemophilus pleuropneumoniae serotyping. J Clin Microbiol. 1982 Nov;16(5):840–843. doi: 10.1128/jcm.16.5.840-843.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SHOPE R. E. PORCINE CONTAGIOUS PLEUROPNEUMONIA. I. EXPERIMENTAL TRANSMISSION, ETIOLOGY, AND PATHOLOGY. J Exp Med. 1964 Mar 1;119:357–368. doi: 10.1084/jem.119.3.357. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SHOPE R. E., WHITE D. C., LEIDY G. PORCINE CONTAGIOUS PLEUROPNEUMONIA. II. STUDIES OF THE PATHOGENICITY OF THE ETIOLOGICAL AGENT, HEMOPHILUS PLEUROPNEUMONIAE. J Exp Med. 1964 Mar 1;119:369–375. doi: 10.1084/jem.119.3.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sebunya T. N., Saunders J. R. Haemophilus pleuropneumoniae infection in swine: a review. J Am Vet Med Assoc. 1983 Jun 15;182(12):1331–1337. [PubMed] [Google Scholar]
- Stenderup J., Orskov F. The clonal nature of enteropathogenic Escherichia coli strains. J Infect Dis. 1983 Dec;148(6):1019–1024. doi: 10.1093/infdis/148.6.1019. [DOI] [PubMed] [Google Scholar]
- Stephens D. S., McGee Z. A. Association of virulence of Neisseria meningitidis with transparent colony type and low-molecular-weight outer membrane proteins. J Infect Dis. 1983 Feb;147(2):282–292. doi: 10.1093/infdis/147.2.282. [DOI] [PubMed] [Google Scholar]
- Swanson J., Kraus S. J., Gotschlich E. C. Studies on gonococcus infection. I. Pili and zones of adhesion: their relation to gonococcal growth patterns. J Exp Med. 1971 Oct 1;134(4):886–906. doi: 10.1084/jem.134.4.886. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Swanson J. Studies on gonococcus infection. XIV. Cell wall protein differences among color/opacity colony variants of Neisseria gonorrhoeae. Infect Immun. 1978 Jul;21(1):292–302. doi: 10.1128/iai.21.1.292-302.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Walstad D. L., Guymon L. F., Sparling P. F. Altered outer membrane protein in different colonial types of Neisseria gonorrhoeae. J Bacteriol. 1977 Mar;129(3):1623–1627. doi: 10.1128/jb.129.3.1623-1627.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]