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Antimicrobial Agents and Chemotherapy, December 2002, p. 3926-3932, Vol. 46, No. 12
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.12.3926-3932.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Energy-Dependent Accumulation of Norfloxacin and Porin Expression in Clinical Isolates of Klebsiella pneumoniae and Relationship to Extended-Spectrum ß-Lactamase Production

Luis Martínez-Martínez,1,2* Alvaro Pascual,1,2 María del Carmen Conejo,1 Isabel García,1 Providencia Joyanes,1 Antonio Doménech-Sánchez,3 and Vicente Javier Benedí3

Department of Microbiology, School of Medicine, University of Seville,1 University Hospital V. Macarena, Seville,2 Laboratory of Microbiology, IMEDEA (CSIC-UIB), Universitat de les Illes Balears, Palma de Mallorca, Spain3

Received 28 January 2002/ Returned for modification 18 May 2002/ Accepted 8 September 2002

The relationships between porin deficiency, active efflux of fluoroquinolones, and extended-spectrum ß-lactamase (ESBL) production were determined for 53 clinical isolates of Klebsiella pneumoniae. Thirty-two ESBL-positive strains (including 22 strains expressing porins and 10 strains lacking porins) and 21 ESBL-negative strains were evaluated. Active efflux of norfloxacin was defined as a >=50% increase in the accumulation of norfloxacin in the presence of carbonyl cyanide m-chlorophenylhydrazone (CCCP) in comparison with the corresponding basal value in the absence of CCCP. The quinolone resistance-determining regions of both gyrA and parC from 13 strains, representing all isolates with different porin profiles and with or without active efflux, were determined. Porin loss was significantly more common among ESBL-positive strains (10 of 32 [31.2%]) than among ESBL-negative strains (0 of 2 [0%]) (P < 0.01). Active efflux was observed in 7 of 10 (70%) strains lacking porins and in 4 of 43 (9.3%) strains producing porins (P < 0.001). The 11 strains showing active efflux corresponded to 3 of 21 (14.3%) ESBL-negative strains and 8 of 32 (25.5%) ESBL-positive strains (P > 0.05). Basal values of norfloxacin accumulation were higher in strains lacking active efflux than in those that had this mechanism (P < 0.05). In the absence of topoisomerase changes, the contribution of either porin loss or active efflux to fluoroquinolone resistance in K. pneumoniae was negligible. It is concluded that among K. pneumoniae strains of clinical origin, porin loss was observed only in those producing ESBL, and that a significant number of porin-deficient strains also expressed active efflux of norfloxacin. In terms of fluoroquinolone resistance, both mechanisms are significant only in the presence of topoisomerase modifications.

* Corresponding author. Mailing address: Department of Microbiology, School of Medicine, University of Seville, Apdo. 914, 41080 Seville, Spain. Phone: 34 95 500 8287. Fax: 34 95 437 7413. E-mail: lmartin{at}us.es.

Antimicrobial Agents and Chemotherapy, December 2002, p. 3926-3932, Vol. 46, No. 12
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.12.3926-3932.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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