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Applied and Environmental Microbiology, June 2003, p. 3359-3367, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3359-3367.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

A Stable Bioluminescent Construct of Escherichia coli O157:H7 for Hazard Assessments of Long-Term Survival in the Environment

Jennifer M. Ritchie,1,2,{dagger} Graeme R. Campbell,1 Jill Shepherd,1 Yvonne Beaton,1 Davey Jones,3 Ken Killham,1 and Rebekka R. E. Artz1*

School of Biological Sciences, University of Aberdeen, Aberdeen AB24 3UU,1 Department of Molecular and Cell Biology, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB25 2ZD, Scotland, and,2 School of Agricultural and Forest Sciences, University of Wales, Bangor LL57 2UW, Wales, United Kingdom3

Received 26 August 2002/ Accepted 18 March 2003

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.

* Corresponding author. Present address: Macaulay Land Use Research Institute, Craigiebuckler, Aberdeen AB15 8QH, Scotland, United Kingdom. Phone: (44) (0)1224 498200. Fax: (44) (0)1224 311556. E-mail: r.artz{at}macaulay.ac.uk.

{dagger} Present address: Division of Geographic Medicine and Infectious Diseases, New England Medical Center, Boston, MA 02111.

Applied and Environmental Microbiology, June 2003, p. 3359-3367, Vol. 69, No. 6
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.6.3359-3367.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.





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