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Applied and Environmental Microbiology, November 2003, p. 6393-6398, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6393-6398.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Assessment of Photodynamic Destruction of Escherichia coli O157:H7 and Listeria monocytogenes by Using ATP Bioluminescence

N. A. Romanova,1,2 L. Y. Brovko,1,2 L. Moore,2 E. Pometun,3 A. P. Savitsky,3,4 N. N. Ugarova,3 and M. W. Griffiths1,2*

Canadian Research Institute for Food Safety,1 Department of Food Science, University of Guelph, Guelph, Ontario, Canada,2 Faculty of Chemistry, Lomonosov Moscow State University,3 Institute of Biochemistry, Russian Academy of Sciences, Moscow, Russia4

Received 26 February 2003/ Accepted 7 August 2003

Antimicrobial photodynamic therapy was shown to be effective against a wide range of bacterial cells, as well as for fungi, yeasts, and viruses. It was shown previously that photodestruction of yeast cells treated with photosensitizers resulted in cell destruction and leakage of ATP. Three photosensitizers were used in this study: tetra(N-methyl-4-pyridyl)porphine tetratosylate salt (TMPyP), toluidine blue O (TBO), and methylene blue trihydrate (MB). A microdilution method was used to determine MICs of the photosensitizers against both Escherichia coli O157:H7 and Listeria monocytogenes. To evaluate the effects of photodestruction on E. coli and L. monocytogenes cells, a bioluminescence method for detection of ATP leakage and a colony-forming assay were used. All tested photosensitizers were effective for photodynamic destruction of both bacteria. The effectiveness of photosensitizers (in microgram-per-milliliter equivalents) decreased in the order TBO > MB > TMPyP for both organisms. The MICs were two- to fourfold higher for E. coli O157:H7 than for L. monocytogenes. The primary effects of all of the photosensitizers tested on live bacterial cells were a decrease in intracellular ATP and an increase in extracellular ATP, accompanied by elimination of viable cells from the sample. The time courses of photodestruction and intracellular ATP leakage were different for E. coli and L. monocytogenes. These results show that bioluminescent ATP-metry can be used for investigation of the first stages of bacterial photodestruction.

* Corresponding author. Mailing address: Canadian Research Institute for Food Safety, University of Guelph, Guelph, ON, Canada N1G 2W1. Phone: (519) 824-4120, ext. 52269. Fax: (519) 763-0952. E-mail: mgriffit{at}uoguelph.ca.

Applied and Environmental Microbiology, November 2003, p. 6393-6398, Vol. 69, No. 11
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.11.6393-6398.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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