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Antimicrobial Agents and Chemotherapy, August 2002, p. 2458-2461, Vol. 46, No. 8
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.8.2458-2461.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Tetracycline Rapidly Reaches All the Constituent Cells of Uropathogenic Escherichia coli Biofilms

G. Stone, P. Wood, L. Dixon, M. Keyhan, and A. Matin*

Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California 94305

Received 3 January 2002/ Returned for modification 8 April 2002/ Accepted 2 May 2002

We have developed a method for visualizing Escherichia coli cells that are exposed to tetracycline in a biofilm, based on a previous report that liposomes containing the E. coli TetR(B) protein fluoresce when exposed to this antibiotic. By our method, cells devoid of TetR(B) also exhibited tetracycline-dependent fluorescence. At 50 µg of tetracycline ml-1, planktonic cells of a uropathogenic E. coli (UPEC) strain developed maximal fluorescence after 7.5 to 10 min of exposure. A similar behavior was exhibited by cells in a 24- or 48-h UPEC biofilm, as examined by confocal laser microscopy, regardless of whether they lined empty spaces or occupied densely packed regions. Further, a comparison of phase-contrast and fluorescent images of corresponding biofilm zones showed that all the cells fluoresced. Thus, all the biofilm cells were exposed to tetracycline and there were no pockets within the biofilm where the antibiotic failed to reach. It also appeared unlikely that niches of reduced exposure to the antibiotic existed within the biofilms.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Stanford University School of Medicine, Sherman Fairchild D317, 299 West Campus Dr., Stanford, CA 94305. Phone: (650) 725-4745. Fax: (650) 725-6757. E-mail: a.matin{at}stanford.edu.

Antimicrobial Agents and Chemotherapy, August 2002, p. 2458-2461, Vol. 46, No. 8
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.8.2458-2461.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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