A stable bioluminescent construct of Escherichia coli O157:H7 for hazard assessments of long-term survival in the environment.

J. M. Ritchie, G. R. Campbell, J. Shepherd, Y. Beaton, D. Jones, Kenneth Stuart Killham, Rebekka Artz

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Abstract

A chromosomally lux-marked (Tn5 luxCDABE) strain of nontoxigenic Escherichia coli O157:H7 was constructed by transposon mutagenesis and shown to have retained the O157, H7, and intimin phenotypes. The survival characteristics of this strain in the experiments performed (soil at -5, -100, and -1,500 kPa matric potential and artificial groundwater) were indistinguishable from the wild-type strain. Evaluation of potential luminescence was found to be a rapid, cheap, and quantitative measure of viable E. coli O157:H7 Tn5 luxCDABE populations in environmental samples. In the survival studies, bioluminescence of the starved populations of E. coli O157:H7 Tn5 luxCDABE could be reactivated to the original levels of light emission, suggesting that these populations remain viable and potentially infective to humans. The attributes of the construct offer a cheap and low-risk substitute to the use of verocytotoxin-producing E. coli O157:H7 in long-term survival studies.

Original languageEnglish
Pages (from-to)3359-3367
Number of pages8
JournalApplied and Environmental Microbiology
Volume69
DOIs
Publication statusPublished - 2003

Keywords

  • MODIFIED PSEUDOMONAS-FLUORESCENS
  • HEMOLYTIC-UREMIC SYNDROME
  • SHIGA TOXIN
  • NONCULTURABLE STATE
  • SOIL
  • LUMINESCENCE
  • BACTERIA
  • WATER
  • STARVATION
  • POPULATION

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