1887

Abstract

Colonization by the commensal strain A0 34/86 (O83 : K24 : H31) has proved to be safe and efficient in the prophylaxis and treatment of nosocomial infections and diarrhoea of preterm and newborn infants in Czech paediatric clinics over the past three decades. In searching for traits contributing to this beneficial effect related to the gut colonization capacity of the strain, the authors have analysed its genome by DNA–DNA hybridization to K-12 (MG1655) genomic DNA arrays and to ‘Pathoarrays’, as well as by multiplex PCR, bacterial artificial chromosome (BAC) library cloning and shotgun sequencing. Four hundred and ten K-12 ORFs were absent from A0 34/86, while 72 out of 456 genes associated with pathogenicity islands of and were also detected in A0 34/86. Furthermore, extraintestinal pathogenic -related genes involved in iron uptake and adhesion were detected by multiplex PCR, and genes encoding the HlyA and cytotoxic necrotizing factor toxins, together with 21 genes of the uropathogenic 536 pathogenicity island II, were identified by analysis of 2304 shotgun and 1344 BAC clone sequences of A0 34/86 DNA. Multiple sequence comparisons identified 31 kb of DNA specific for A0 34/86; some of the genes carried by this DNA may prove to be implicated in the colonization capacity of the strain, enabling it to outcompete pathogens. Among 100 examined BAC clones roughly covering the A0 34/86 genome, one reproducibly conferred on the laboratory strain DH10B an enhanced capacity to persist in the intestine of newborn piglets. Sequencing revealed that this BAC clone carried gene clusters encoding gluconate and mannonate metabolism, adhesion (), invasion () and restriction/modification functions. Hence, the genome of this clinically safe and highly efficient colonizer strain appears to harbour many ‘virulence-associated’ genes. These results highlight the thin line between bacterial ‘virulence’ and ‘fitness' or ‘colonization’ factors, and question the definition of enterobacterial virulence factors.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.27469-0
2005-02-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/151/2/mic1510385.html?itemId=/content/journal/micro/10.1099/mic.0.27469-0&mimeType=html&fmt=ahah

References

  1. Altschul S. F., Madden T. L., Schaffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. 1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402 [CrossRef]
    [Google Scholar]
  2. Bach S., de Almeida A., Carniel E. 2000; The Yersinia high-pathogenicity island is present in different members of the family Enterobacteriaceae. FEMS Microbiol Lett 183:289–294 [CrossRef]
    [Google Scholar]
  3. Baga M., Goransson M., Normark S., Uhlin B. E. 1985; Transcriptional activation of a pap pilus virulence operon from uropathogenic Escherichia coli. EMBO J 4:3887–3893
    [Google Scholar]
  4. Blattner F. R., Plunkett G. R., Bloch C. A. & 14 other authors; 1997; The complete genome sequence of Escherichia coli K-12. Science 277:1453–1474 [CrossRef]
    [Google Scholar]
  5. Bonacorsi S., Clermont O., Houdouin V. & 7 other authors; 2003; Molecular analysis and experimental virulence of French and North American Escherichia coli neonatal meningitis isolates: identification of a new virulent clone. J Infect Dis 187:1895–1906 [CrossRef]
    [Google Scholar]
  6. Buchrieser C., Weagant S. D., Kaspar C. W. 1994; Molecular characterization of Yersinia enterocolitica by pulsed-field gel electrophoresis and hybridization of DNA fragments to ail and pYV probes. Appl Environ Microbiol 60:4371–4379
    [Google Scholar]
  7. Buchrieser C., Brosch R., Bach S., Guiyoule A., Carniel E. 1998; The high-pathogenicity island of Yersinia pseudotuberculosis can be inserted into any of the three chromosomal asn tRNA genes. Mol Microbiol 30:965–978 [CrossRef]
    [Google Scholar]
  8. Buchrieser C., Rusniok C., Frangeul L., Couve E., Billault A., Kunst F., Carniel E., Glaser P. 1999; The 102-kilobase pgm locus of Yersinia pestis: sequence analysis and comparison of selected regions among differentYersinia pestis and Yersinia pseudotuberculosis strains. Infect Immun 67:4851–4861
    [Google Scholar]
  9. Carniel E., Guilvout I., Prentice M. 1996; Characterization of a large chromosomal “high-pathogenicity island” in biotype 1B Yersinia enterocolitica . J Bacteriol 178:6743–6751
    [Google Scholar]
  10. Clermont O., Bonacorsi S., Bingen E. 2000; Rapid and simple determination of the Escherichia coli phylogenetic group. Appl Environ Microbiol 66:4555–4558 [CrossRef]
    [Google Scholar]
  11. De Rycke J., Nougayrede J. P., Oswald E., Mazars P. 1997; Interaction of Escherichia coli producing cytotoxic necrotizing factor with HeLa epithelial cells. Adv Exp Med Biol 412:363–366
    [Google Scholar]
  12. Dobrindt U., Blum-Oehler G., Nagy G., Schneider G., Johann A., Gottschalk G., Hacker J. 2002; Genetic structure and distribution of four pathogenicity islands (PAI I(536) to PAI IV(536)) of uropathogenicEscherichia coli strain 536. Infect Immun 70:6365–6372 [CrossRef]
    [Google Scholar]
  13. Dobrindt U., Agerer F., Michaelis K. & 7 other authors; 2003; Analysis of genome plasticity in pathogenic and commensal Escherichia coli isolates by use of DNA arrays. J Bacteriol 185:1831–1840 [CrossRef]
    [Google Scholar]
  14. Dozois C. M., Dho-Moulin M., Bree A., Fairbrother J. M., Desautels C., Curtiss R. 3rd (2000; Relationship between the Tsh autotransporter and pathogenicity of avian Escherichia coli and localization and analysis of the Tsh genetic region. Infect Immun 68:4145–4154 [CrossRef]
    [Google Scholar]
  15. Ewing B., Green P. 1998; Base-calling of automated sequencer traces using phred. II. Error probabilities. Genome Res 8:186–194
    [Google Scholar]
  16. Frangeul L., Glaser P., Rusniok C., Buchrieser C., Duchaud E., Dehoux P., Kunst F. 2004; CAAT-Box, contigs-Assembly and Annotation Tool-Box for genome sequencing projects. Bioinformatics 20:790–797 [CrossRef]
    [Google Scholar]
  17. Gilson L., Mahanty H. K., Kolter R. 1990; Genetic analysis of an MDR-like export system: the secretion of colicin V. EMBO J 9:3875–3894
    [Google Scholar]
  18. Gordon D., Abajian C., Green P. 1998; Consed: a graphical tool for sequence finishing. Genome Res 8:195–202 [CrossRef]
    [Google Scholar]
  19. Grozdanov L., Raasch C., Schulze J., Sonnenborn U., Gottschalk G., Hacker J., Dobrindt U. 2004; Analysis of the genome structure of the non-pathogenic probiotic Escherichia coli Strain NIssle 1917. J Bacteriol 186:5432–5441 [CrossRef]
    [Google Scholar]
  20. Hacker J., Carniel E. 2001; Ecological fitness, genomic islands and bacterial pathogenicity. A Darwinian view of the evolution of microbes. EMBO Rep 2:376–381 [CrossRef]
    [Google Scholar]
  21. Hayashi T., Makino K., Ohnishi M. & 19 other authors; 2001; Complete genome sequence of enterohemorrhagic Escherichia coli O157 : H7 and genomic comparison with a laboratory strain K-12. DNA Res 8:11–22 [CrossRef]
    [Google Scholar]
  22. Huang S. H., Wass C., Fu Q., Prasadarao N. V., Stins M., Kim K. S. 1995; Escherichia coli invasion of brain microvascular endothelial cells in vitro and in vivo: molecular cloning and characterization of invasion gene ibe10. Infect Immun 63:4470–4475
    [Google Scholar]
  23. Huang S. H., Chen Y. H., Kong G., Chen S. H., Besemer J., Borodovsky M., Jong A. 2001; A novel genetic island of meningitic Escherichia coli K1 containing the ibeA invasion gene (GimA): functional annotation and carbon-source-regulated invasion of human brain microvascular endothelial cells. Funct Integr Genomics 1:312–322 [CrossRef]
    [Google Scholar]
  24. Johnson J. R., Stell A. L. 2000; Extended virulence genotypes of Escherichia coli strains from patients with urosepsis in relation to phylogeny and host compromise. J Infect Dis 181:261–272 [CrossRef]
    [Google Scholar]
  25. Kaper J. B., Hacker J. 1999; Pathogenicity Islands and Other Mobile Genetic Elements of Diarrheagenic Escherichia coli. Chapter 3 in Pathogenicity Islands and Other Mobile Virulence Elements pp 33–58 Washington, DC: American Society for Microbiology;
    [Google Scholar]
  26. Lan R., Reeves P. R. 2000; Intraspecies variation in bacterial genomes: the need for a species genome concept. Trends Microbiol 9:396–401
    [Google Scholar]
  27. Lodinova R., Jouja V. 1977; Antibody production by the mammary gland in mothers after artificial oral colonisation of their infants with a non-pathogenic strain E. coli O83. Acta Paediatr Scand 66:705–708 [CrossRef]
    [Google Scholar]
  28. Lodinova R., Jouja V., Lanc A. 1967; Influence of the intestinal flora on the development of immune reactions in infants. J Bacteriol 93:797–800
    [Google Scholar]
  29. Lodinova R., Jouja V., Vinsova N., Vocel J., Melkova J. 1980; New attempts and possibilities in prevention and treatment of intestinal coli-infections in infants. Czech Med 3:47–58
    [Google Scholar]
  30. Lodinova-Zadnikova R., Slavikova M., Tlaskalova-Hogenova H., Adlerberth I., Hanson L. A., Wold A., Carlsson B., Svanborg C., Mellander L. 1991; The antibody response in breast-fed and non-breast-fed infants after artificial colonization of the intestine with Escherichia coli O83. Pediatr Res 29:396–399 [CrossRef]
    [Google Scholar]
  31. Lodinova-Zadnikova R., Tlaskalova H., Korych B., Bartakova Z. 1995; The antibody response in infants after oral administration of inactivated and living E. coli vaccines and their protective effect against nosocomial infections. Adv Exp Med Biol 371B:1431–1438
    [Google Scholar]
  32. Lodinova-Zadnikova R., Sonnenborn U., Tlaskalova H. 1998; Probiotics and E. coli infections in man. Vet Q 20:S78–81 [CrossRef]
    [Google Scholar]
  33. Lodinova-Zadnikova R., Cukrowska B., Tlaskalova-Hogenova H. 2003; Oral administration of probiotic Escherichia coli after birth reduces frequency of allergies and repeated infections later in life (after 10 and 20 years. Int Arch Allergy Immunol 131:209–211 [CrossRef]
    [Google Scholar]
  34. Nataro J. P., Kaper J. B. 1998; Diarrheagenic Escherichia coli. Clin Microbiol Rev 11:142–201
    [Google Scholar]
  35. Ochman H., Jones I. B. 2000; Evolutionary dynamics of full genome content in Escherichia coli. EMBO J 19:6637–6643 [CrossRef]
    [Google Scholar]
  36. Otto B. R., van Dooren S. J., Nuijens J. H., Luirink J., Oudega B. 1998; Characterization of a hemoglobin protease secreted by the pathogenic Escherichia coli strain EB1. J Exp Med 188:1091–1103 [CrossRef]
    [Google Scholar]
  37. Pearson W. R. 1990; Rapid and sensitive sequence comparison with fastp and fasta. Methods Enzymol 183:63–98
    [Google Scholar]
  38. Pearson W. R., Lipman D. J. 1988; Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A 85:2444–2448 [CrossRef]
    [Google Scholar]
  39. Perna N. T., Plunkett G., Burland V. 25 other authors 2001; Genome sequence of enterohaemorrhagic Escherichia coli O157 : H7. Nature 409:529–533 [CrossRef]
    [Google Scholar]
  40. Rodriguez E., Gaggero C., Lavina M. 1999; The structural gene for microcin H47 encodes a peptide precursor with antibiotic activity. Antimicrob Agents Chemother 43:2176–2182
    [Google Scholar]
  41. Roe A. J., Currie C., Smith D. G. E., Gally D. L. 2001; Analysis of type 1 fimbriae expression in verotoxigenic Escherichia coli: a comparison between serotypes O157 and O26. Microbiology 147:145–152
    [Google Scholar]
  42. Rutherford K., Parkhill J., Crook J., Horsnell T., Rice P., Rajandream M. A., Barrell B. 2000; Artemis: sequence visualization and annotation. Bioinformatics 16:944–955 [CrossRef]
    [Google Scholar]
  43. Welch R. A. 2001; RTX toxin structure and function: a story of numerous anomalies and few analogies in toxin biology. Curr Top Microbiol Immunol 257:85–111
    [Google Scholar]
  44. Welch R. A., Burland V., Plunkett G. 16 other authors 3rd & 2002; Extensive mosaic structure revealed by the complete genome sequence of uropathogenic Escherichia coli . Proc Natl Acad Sci U S A 99:17020–17024 [CrossRef]
    [Google Scholar]
  45. Weiserova M., Janscak P., Benada O., Hubacek J., Zinkevich V. E., Glover S. W., Firman K. 1993; Cloning, production and characterisation of wild type and mutant forms of the R.EcoK endonucleases. Nucleic Acids Res 21:373–379 [CrossRef]
    [Google Scholar]
  46. Wold A. E., Caugant D. A., Lidin-Janson G., de Man P., Svanborg C. 1992; Resident colonic Escherichia coli strains frequently display uropathogenic characteristics. J Infect Dis 165:46–52 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.27469-0
Loading
/content/journal/micro/10.1099/mic.0.27469-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Supplementary material 2

PDF

Supplementary material 3

PDF
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