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. 1997 Jun;35(6):1550–1555. doi: 10.1128/jcm.35.6.1550-1555.1997

Molecular characterization of two Bordetella bronchiseptica strains isolated from children with coughs.

P Stefanelli 1, P Mastrantonio 1, S Z Hausman 1, M Giuliano 1, D L Burns 1
PMCID: PMC229785  PMID: 9163480

Abstract

During a surveillance program associated with the Italian clinical trial for the evaluation of new acellular pertussis vaccines, two bacterial isolates were obtained in cultures of samples from immunocompetent infants who had episodes of cough. Both clinical isolates were identified as Bordetella bronchiseptica by biochemical criteria, although both strains agglutinated with antisera specific for Bordetella parapertussis, suggesting that the strains exhibited some characteristics of both B. bronchiseptica and B. parapertussis. Both children from whom these strains were isolated exhibited an increase in serum antibody titer to pertussis toxin (PT), a protein that is produced by Bordetella pertussis but that is not thought to be produced by B. bronchiseptica. We therefore examined whether the clinical isolates were capable of producing PT. Neither strain produced PT under laboratory conditions, although both strains appeared to contain a portion of the ptx region that encodes the structural subunits of PT. In order to determine whether the ptx genes may encode functional proteins, we inserted an active promoter directly upstream of the ptx region of one of these strains. Biologically active PT was produced, suggesting that this strain contains the genetic information necessary to encode an active PT molecule. Sequence analysis of the ptx promoter region of both strains indicated that, while they shared homology with the B. bronchiseptica ATCC 4617 sequence, they contained certain sequence motifs that are characteristic of B. parapertussis and certain motifs that are characteristic of B. pertussis. Taken together, these findings suggest that variant strains of B. bronchiseptica exist and might be capable of causing significant illness in humans.

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Selected References

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  1. Aricò B., Rappuoli R. Bordetella parapertussis and Bordetella bronchiseptica contain transcriptionally silent pertussis toxin genes. J Bacteriol. 1987 Jun;169(6):2847–2853. doi: 10.1128/jb.169.6.2847-2853.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brennan M. J., Li Z. M., Cowell J. L., Bisher M. E., Steven A. C., Novotny P., Manclark C. R. Identification of a 69-kilodalton nonfimbrial protein as an agglutinogen of Bordetella pertussis. Infect Immun. 1988 Dec;56(12):3189–3195. doi: 10.1128/iai.56.12.3189-3195.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Camilli A., Mekalanos J. J. Use of recombinase gene fusions to identify Vibrio cholerae genes induced during infection. Mol Microbiol. 1995 Nov;18(4):671–683. doi: 10.1111/j.1365-2958.1995.mmi_18040671.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cotter P. A., Miller J. F. BvgAS-mediated signal transduction: analysis of phase-locked regulatory mutants of Bordetella bronchiseptica in a rabbit model. Infect Immun. 1994 Aug;62(8):3381–3390. doi: 10.1128/iai.62.8.3381-3390.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Frank D. W., Parker C. D. Interaction of monoclonal antibodies with pertussis toxin and its subunits. Infect Immun. 1984 Oct;46(1):195–201. doi: 10.1128/iai.46.1.195-201.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gentry-Weeks C. R., Cookson B. T., Goldman W. E., Rimler R. B., Porter S. B., Curtiss R., 3rd Dermonecrotic toxin and tracheal cytotoxin, putative virulence factors of Bordetella avium. Infect Immun. 1988 Jul;56(7):1698–1707. doi: 10.1128/iai.56.7.1698-1707.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gillenius P., Jätmaa E., Askelöf P., Granström M., Tiru M. The standardization of an assay for pertussis toxin and antitoxin in microplate culture of Chinese hamster ovary cells. J Biol Stand. 1985 Jan;13(1):61–66. doi: 10.1016/s0092-1157(85)80034-2. [DOI] [PubMed] [Google Scholar]
  8. Goodnow R. A. Biology of Bordetella bronchiseptica. Microbiol Rev. 1980 Dec;44(4):722–738. doi: 10.1128/mr.44.4.722-738.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Greco D., Salmaso S., Mastrantonio P., Giuliano M., Tozzi A. E., Anemona A., Ciofi degli Atti M. L., Giammanco A., Panei P., Blackwelder W. C. A controlled trial of two acellular vaccines and one whole-cell vaccine against pertussis. Progetto Pertosse Working Group. N Engl J Med. 1996 Feb 8;334(6):341–348. doi: 10.1056/NEJM199602083340601. [DOI] [PubMed] [Google Scholar]
  10. Hausman S. Z., Cherry J. D., Heininger U., Wirsing von König C. H., Burns D. L. Analysis of proteins encoded by the ptx and ptl genes of Bordetella bronchiseptica and Bordetella parapertussis. Infect Immun. 1996 Oct;64(10):4020–4026. doi: 10.1128/iai.64.10.4020-4026.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Heininger U., Stehr K., Schmitt-Grohé S., Lorenz C., Rost R., Christenson P. D., Uberall M., Cherry J. D. Clinical characteristics of illness caused by Bordetella parapertussis compared with illness caused by Bordetella pertussis. Pediatr Infect Dis J. 1994 Apr;13(4):306–309. doi: 10.1097/00006454-199404000-00011. [DOI] [PubMed] [Google Scholar]
  12. Hewlett E. L., Gordon V. M., McCaffery J. D., Sutherland W. M., Gray M. C. Adenylate cyclase toxin from Bordetella pertussis. Identification and purification of the holotoxin molecule. J Biol Chem. 1989 Nov 15;264(32):19379–19384. [PubMed] [Google Scholar]
  13. Hewlett E. L., Sauer K. T., Myers G. A., Cowell J. L., Guerrant R. L. Induction of a novel morphological response in Chinese hamster ovary cells by pertussis toxin. Infect Immun. 1983 Jun;40(3):1198–1203. doi: 10.1128/iai.40.3.1198-1203.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Idigbe E. O., Parton R., Wardlaw A. C. Rapidity of antigenic modulation of Bordetella pertussis in modified Hornibrook medium. J Med Microbiol. 1981 Nov;14(4):409–418. doi: 10.1099/00222615-14-4-409. [DOI] [PubMed] [Google Scholar]
  15. JONES M. Subacute bacterial endocarditis of nonstreptococcic etiology; a review of the literature of the thirteen-year period 1936-1948 inclusive. Am Heart J. 1950 Jul;40(1):106–116. doi: 10.1016/0002-8703(50)90154-2. [DOI] [PubMed] [Google Scholar]
  16. Johnson F. D., Burns D. L. Detection and subcellular localization of three Ptl proteins involved in the secretion of pertussis toxin from Bordetella pertussis. J Bacteriol. 1994 Sep;176(17):5350–5356. doi: 10.1128/jb.176.17.5350-5356.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kenimer J. G., Kim K. J., Probst P. G., Manclark C. R., Burstyn D. G., Cowell J. L. Monoclonal antibodies to pertussis toxin: utilization as probes of toxin function. Hybridoma. 1989 Feb;8(1):37–51. doi: 10.1089/hyb.1989.8.37. [DOI] [PubMed] [Google Scholar]
  18. Khattak M. N., Matthews R. C. Genetic relatedness of Bordetella species as determined by macrorestriction digests resolved by pulsed-field gel electrophoresis. Int J Syst Bacteriol. 1993 Oct;43(4):659–664. doi: 10.1099/00207713-43-4-659. [DOI] [PubMed] [Google Scholar]
  19. LACEY B. W. Antigenic modulation of Bordetella pertussis. J Hyg (Lond) 1960 Mar;58:57–93. doi: 10.1017/s0022172400038134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Leininger E., Probst P. G., Brennan M. J., Kenimer J. G. Inhibition of Bordetella pertussis filamentous hemagglutinin-mediated cell adherence with monoclonal antibodies. FEMS Microbiol Lett. 1993 Jan 1;106(1):31–38. doi: 10.1111/j.1574-6968.1993.tb05931.x. [DOI] [PubMed] [Google Scholar]
  22. Locht C., Bertin P., Menozzi F. D., Renauld G. The filamentous haemagglutinin, a multifaceted adhesion produced by virulent Bordetella spp. Mol Microbiol. 1993 Aug;9(4):653–660. doi: 10.1111/j.1365-2958.1993.tb01725.x. [DOI] [PubMed] [Google Scholar]
  23. Marchitto K. S., Munoz J. J., Keith J. M. Detection of subunits of pertussis toxin in Tn5-induced Bordetella mutants deficient in toxin biological activity. Infect Immun. 1987 May;55(5):1309–1313. doi: 10.1128/iai.55.5.1309-1313.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mastrantonio P., Stefanelli P., Giuliano M. Polymerase chain reaction for the detection of Bordetella pertussis in clinical nasopharyngeal aspirates. J Med Microbiol. 1996 Apr;44(4):261–266. doi: 10.1099/00222615-44-4-261. [DOI] [PubMed] [Google Scholar]
  25. Meade B. D., Deforest A., Edwards K. M., Romani T. A., Lynn F., O'Brien C. H., Swartz C. B., Reed G. F., Deloria M. A. Description and evaluation of serologic assays used in a multicenter trial of acellular pertussis vaccines. Pediatrics. 1995 Sep;96(3 Pt 2):570–575. [PubMed] [Google Scholar]
  26. Meis J. F., van Griethuijsen A. J., Muytjens H. L. Bordetella bronchiseptica bronchitis in an immunosuppressed patient. Eur J Clin Microbiol Infect Dis. 1990 May;9(5):366–367. doi: 10.1007/BF01973748. [DOI] [PubMed] [Google Scholar]
  27. Mesnard R., Guiso N., Michelet C., Sire J. M., Pouëdras P., Donnio P. Y., Avril J. L. Isolation of Bordetella bronchiseptica from a patient with AIDS. Eur J Clin Microbiol Infect Dis. 1993 Apr;12(4):304–306. doi: 10.1007/BF01967267. [DOI] [PubMed] [Google Scholar]
  28. Nicosia A., Perugini M., Franzini C., Casagli M. C., Borri M. G., Antoni G., Almoni M., Neri P., Ratti G., Rappuoli R. Cloning and sequencing of the pertussis toxin genes: operon structure and gene duplication. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4631–4635. doi: 10.1073/pnas.83.13.4631. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. SHIH MAN C. Pertussis due to Brucella bronchoseptica; case report. Pediatrics. 1950 Aug;6(2):227–228. [PubMed] [Google Scholar]
  30. Stefanelli P., Giuliano M., Bottone M., Spigaglia P., Mastrantonio P. Polymerase chain reaction for the identification of Bordetella pertussis and Bordetella parapertussis. Diagn Microbiol Infect Dis. 1996 Apr;24(4):197–200. doi: 10.1016/0732-8893(96)00064-8. [DOI] [PubMed] [Google Scholar]
  31. Weiss A. A., Melton A. R., Walker K. E., Andraos-Selim C., Meidl J. J. Use of the promoter fusion transposon Tn5 lac to identify mutations in Bordetella pertussis vir-regulated genes. Infect Immun. 1989 Sep;57(9):2674–2682. doi: 10.1128/iai.57.9.2674-2682.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Woolfrey B. F., Moody J. A. Human infections associated with Bordetella bronchiseptica. Clin Microbiol Rev. 1991 Jul;4(3):243–255. doi: 10.1128/cmr.4.3.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. de la Fuente J., Albo C., Rodríguez A., Sopeña B., Martínez C. Bordetella bronchiseptica pneumonia in a patient with AIDS. Thorax. 1994 Jul;49(7):719–720. doi: 10.1136/thx.49.7.719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. van der Zee A., Agterberg C., van Agterveld M., Peeters M., Mooi F. R. Characterization of IS1001, an insertion sequence element of Bordetella parapertussis. J Bacteriol. 1993 Jan;175(1):141–147. doi: 10.1128/jb.175.1.141-147.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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