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Applied and Environmental Microbiology, November 2002, p. 5737-5740, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5737-5740.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Sensitive In Situ Monitoring of a Recombinant Bioluminescent Yersinia enterocolitica Reporter Mutant in Real Time on Camembert Cheese

Ariel Maoz,¹ Ralf Mayr,¹ Geraldine Bresolin,¹ Klaus Neuhaus,¹ Kevin P. Francis,² and Siegfried Scherer^1*

Institut für Mikrobiologie, FML-Weihenstephan, Technische Universität München, D-85350 Freising, Germany,¹ Xenogen Corporation, Alameda, California 94501²

Received 22 March 2002/ Accepted 5 August 2002

Bioluminescent mutants of Yersinia enterocolitica were generated by transposon mutagenesis using a promoterless, complete lux operon (luxCDABE) derived from Photorhabdus luminescens, and their production of light in the cheese environment was monitored. Mutant B94, which had the lux cassette inserted into an open reading frame of unknown function was used for direct monitoring of Y. enterocolitica cells on cheeses stored at 10°C by quantifying bioluminescence using a photon-counting, intensified charge-coupled device camera. The detection limit on cheese was 200 CFU/cm². Bioluminescence of the reporter mutant was significantly regulated by its environment (NaCl, temperature, and cheese), as well as by growth phase, via the promoter the lux operon had acquired upon transposition. At low temperatures, mutant B94 did not exhibit the often-reported decrease of photon emission in older cells. It was not necessary to include either antibiotics or aldehyde in the food matrix in order to gain quantitative, reproducible bioluminescence data. As far as we know, this is the first time a pathogen has been monitored in situ, in real time, in a "real-product" status, and at a low temperature.

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* Corresponding author. Mailing address: Institut für Mikrobiologie, FML-Weihenstephan, Technische Universität München, D-85350 Freising, Germany. Phone: 49 8161 713516. Fax: 49 8161 714512. E-mail: Siegfried.Scherer{at}lrz.tum.de.

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Applied and Environmental Microbiology, November 2002, p. 5737-5740, Vol. 68, No. 11
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.11.5737-5740.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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