1887

Abstract

The conjugative plasmid pOLA52, which confers resistance to olaquindox and other antimicrobial agents through a multidrug efflux pump, was investigated for its ability to promote biofilm formation in . Screening of a transposon-mutagenized pOLA52 clone library revealed several biofilm-deficient mutants, which all mapped within a putative operon with high homology to the operon of , where these genes are responsible for type 3 fimbriae expression, attachment to surfaces and biofilm formation. Biofilm formation in microtitre plates and in urinary catheters of clones containing pOLA52 with a disrupted putative operon was reduced by more than 100-fold and 2-fold, respectively, compared to mutants with an intact operon. The conjugative transfer rate of pOLA52 was also significantly lower when the operon was disrupted. Through reverse transcriptase analysis, it was demonstrated that the genes contained in the putative operon were linked and likely to be expressed as a single operon. Immunoblotting with type 3 fimbriae (MrkA)-specific antibodies further verified expression of type 3 fimbriae. When transferred to other, potentially pathogenic, members of the family , including , Typhimurium, sp. and , pOLA52 facilitated increased biofilm formation. pOLA52 is believed to represent the first example of a conjugative plasmid encoding type 3 fimbriae, resulting in enhanced conjugation frequencies and biofilm formation of the plasmid-harbouring strain.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.2007/010454-0
2008-01-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/154/1/187.html?itemId=/content/journal/micro/10.1099/mic.0.2007/010454-0&mimeType=html&fmt=ahah

References

  1. Adegbola R. A., Old D. C. 1983; Fimbrial haemagglutinins in Enterobacter species. J Gen Microbiol 129:2175–2180
    [Google Scholar]
  2. Allen B. L., Gerlach G. F., Clegg S. 1991; Nucleotide sequence and functions of mrk determinants necessary for expression of type 3 fimbriae in Klebsiella pneumoniae . J Bacteriol 173:916–920
    [Google Scholar]
  3. Brunder W., Khan A. S., Hacker J., Karch H. 2001; Novel type of fimbriae encoded by the large plasmid of sorbitol-fermenting enterohemorrhagic Escherichia coli O157 : H(–). Infect Immun 69:4447–4457
    [Google Scholar]
  4. Burmølle M., Hansen L. H., Sørensen S. J. 2005; Use of a whole-cell biosensor and flow cytometry to detect AHL production by an indigenous soil community during decomposition of litter. Microb Ecol 50:221–229
    [Google Scholar]
  5. Burmølle M., Webb J. S., Rao D., Hansen L. H., Sørensen S. J., Kjelleberg S. 2006; Enhanced biofilm formation and increased resistance towards antimicrobial agents and bacterial invasion are caused by synergistic interactions in multispecies biofilms. Appl Environ Microbiol 72:3916–3923
    [Google Scholar]
  6. Castonguay M. H., van der Schaaf S., Koester W., Krooneman J., van der Meer W., Harmsen H., Landini P. 2006; Biofilm formation by Escherichia coli is stimulated by synergistic interactions and co-adhesion mechanisms with adherence-proficient bacteria. Res Microbiol 157:471–478
    [Google Scholar]
  7. de Lipthay J. R., Barkay T., Sørensen S. J. 2001; Enhanced degradation of phenoxyacetic acid in soil by horizontal transfer of the tfdA gene encoding a 2,4-dichlorophenoxyacetic acid dioxygenase. FEMS Microbiol Ecol 35:75–84
    [Google Scholar]
  8. Di Martino P., Cafferini N., Joly B., Darfeuille-Michaud A. 2003; Klebsiella pneumoniae type 3 pili facilitate adherence and biofilm formation on abiotic surfaces. Res Microbiol 154:9–16
    [Google Scholar]
  9. Donlan R. M., Costerton J. W. 2002; Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 15:167–193
    [Google Scholar]
  10. Dudley E. G., Abe C., Ghigo J. M., Latour-Lambert P., Hormazabal J. C., Nataro J. P. 2006; An IncI1 plasmid contributes to the adherence of the atypical enteroaggregative Escherichia coli strain C1096 to cultured cells and abiotic surfaces. Infect Immun 74:2102–2114
    [Google Scholar]
  11. Fux C. A., Costerton J. W., Stewart P. S., Stoodley P. 2005; Survival strategies of infectious biofilms. Trends Microbiol 13:34–40
    [Google Scholar]
  12. Gerlach G. F., Allen B. L., Clegg S. 1988; Molecular characterization of the type 3 (MR/K) fimbriae of Klebsiella pneumoniae . J Bacteriol 170:3547–3553
    [Google Scholar]
  13. Ghigo J. M. 2001; Natural conjugative plasmids induce bacterial biofilm development. Nature 412:442–445
    [Google Scholar]
  14. Hansen L. H., Sørensen S. J., Jensen L. B. 1997; Chromosomal insertion of the entire Escherichia coli lactose operon, into two strains of Pseudomonas , using a modified mini-Tn5 delivery system. Gene 186:167–173
    [Google Scholar]
  15. Hansen L. H., Johannesen E., Burmølle M., Sørensen A. H., Sørensen S. J. 2004; Plasmid-encoded multidrug efflux pump conferring resistance to olaquindox in Escherichia coli . Antimicrob Agents Chemother 48:3332–3337
    [Google Scholar]
  16. Hansen L. H., Sørensen S. J., Jørgensen H. S., Jensen L. B. 2005; The prevalence of the OqxAB multidrug efflux pump amongst olaquindox-resistant Escherichia coli in pigs. Microb Drug Resist 11:378–382
    [Google Scholar]
  17. Hansen L. H., Jensen L. B., Sørensen H. I., Sørensen S. J. 2007; Substrate specificity of the OqxAB multidrug resistance pump in Escherichia coli and selected enteric bacteria. J Antimicrob Chemother 60:145–147
    [Google Scholar]
  18. Hornick D. B., Allen B. L., Horn M. A., Clegg S. 1991; Fimbrial types among respiratory isolates belonging to the family Enterobacteriaceae . J Clin Microbiol 29:1795–1800
    [Google Scholar]
  19. Jagnow J., Clegg S. 2003; Klebsiella pneumoniae MrkD-mediated biofilm formation on extracellular matrix- and collagen-coated surfaces. Microbiology 149:2397–2405
    [Google Scholar]
  20. Jefferson K. K. 2004; What drives bacteria to produce a biofilm?. FEMS Microbiol Lett 236:163–173
    [Google Scholar]
  21. Klausen M., Gjermansen M., Kreft J. U., Tolker-Nielsen T. 2006; Dynamics of development and dispersal in sessile microbial communities: examples from Pseudomonas aeruginosa and Pseudomonas putida model biofilms. FEMS Microbiol Lett 261:1–11
    [Google Scholar]
  22. Langstraat J., Bohse M., Clegg S. 2001; Type 3 fimbrial shaft (MrkA) of Klebsiella pneumoniae , but not the fimbrial adhesin (MrkD), facilitates biofilm formation. Infect Immun 69:5805–5812
    [Google Scholar]
  23. Levine M. M. 1987; Escherichia coli that cause diarrhea: enterotoxigenic, enteropathogenic, enteroinvasive, enterohemorrhagic, and enteroadherent. J Infect Dis 155:377–389
    [Google Scholar]
  24. Livrelli V., De Champs C., Di Martino P., Darfeuille-Michaud A., Forestier C., Joly B. 1996; Adhesive properties and antibiotic resistance of Klebsiella, Enterobacter , and Serratia clinical isolates involved in nosocomial infections. J Clin Microbiol 34:1963–1969
    [Google Scholar]
  25. Miller J. H. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  26. O'Toole G. A., Kolter R. 1998; Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. Mol Microbiol 28:449–461
    [Google Scholar]
  27. Old D. C., Adegbola R. A. 1985; Antigenic relationships among type-3 fimbriae of Enterobacteriaceae revealed by immunoelectronmicroscopy. J Med Microbiol 20:113–121
    [Google Scholar]
  28. Olsen J. E., Brown D. J., Thomsen L. E., Platt D. J., Chadfield M. S. 2004; Differences in the carriage and the ability to utilize the serotype associated virulence plasmid in strains of Salmonella enterica serotype Typhimurium investigated by use of a self-transferable virulence plasmid, pOG669. Microb Pathog 36:337–347
    [Google Scholar]
  29. Reisner A., Haagensen J. A., Schembri M. A., Zechner E. L., Molin S. 2003; Development and maturation of Escherichia coli K-12 biofilms. Mol Microbiol 48:933–946
    [Google Scholar]
  30. Reisner A., Holler B. M., Molin S., Zechner E. L. 2006; Synergistic effects in mixed Escherichia coli biofilms: conjugative plasmid transfer drives biofilm expansion. J Bacteriol 188:3582–3588
    [Google Scholar]
  31. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  32. Sauer F. G., Barnhart M., Choudhury D., Knight S. D., Waksman G., Hultgren S. J. 2000; Chaperone-assisted pilus assembly and bacterial attachment. Curr Opin Struct Biol 10:548–556
    [Google Scholar]
  33. Schurtz T. A., Hornick D. B., Korhonen T. K., Clegg S. 1994; The type 3 fimbrial adhesin gene ( mrkD ) of Klebsiella species is not conserved among all fimbriate strains. Infect Immun 62:4186–4191
    [Google Scholar]
  34. Silhavy T. J., Berman M. L., Enquist L. W. 1984 Experiments with Gene Fusion Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  35. Sørensen A. H., Hansen L. H., Johannesen E., Sørensen S. J. 2003; Conjugative plasmid conferring resistance to olaquindox. Antimicrob Agents Chemother 47:798–799
    [Google Scholar]
  36. Sutherland I. 2001; Biofilm exopolysaccharides: a strong and sticky framework. Microbiology 147:3–9
    [Google Scholar]
  37. Tarkkanen A. M., Virkola R., Clegg S., Korhonen T. K. 1997; Binding of the type 3 fimbriae of Klebsiella pneumoniae to human endothelial and urinary bladder cells. Infect Immun 65:1546–1549
    [Google Scholar]
  38. Vila J., Vargas M., Henderson I. R., Gascon J., Nataro J. P. 2000; Enteroaggregative Escherichia coli virulence factors in traveler's diarrhea strains. J Infect Dis 182:1780–1783
    [Google Scholar]
  39. Williams P., Tomas J. M. 1990; The pathogenicity of Klebsiella pneumoniae . Rev Med Microbiol 1:196–204
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.2007/010454-0
Loading
/content/journal/micro/10.1099/mic.0.2007/010454-0
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