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A Cystic Fibrosis Epidemic Strain of Pseudomonas aeruginosa Displays Enhanced Virulence and Antimicrobial Resistance

Prabhakar Salunkhe,^1, Catherine H. M. Smart,² J. Alun W. Morgan,³ Stavroula Panagea,² Martin J. Walshaw,⁴ C. Anthony Hart,² Robert Geffers,⁵ Burkhard Tümmler,¹ and Craig Winstanley^2*

Clinical Research Group OE 6711, Medical Research School Hannover, 30625 Hannover, Germany,¹ Division of Medical Microbiology, University of Liverpool, Liverpool L69 3GA, United Kingdom,² Warwick-HRI, University of Warwick, Wellesbourne, Warwickshire CV35 9EF, United Kingdom,³ Regional Adult Cystic Fibrosis Unit, Cardiothoracic Center, Liverpool L14 3PE, United Kingdom,⁴ German Research Center for Biotechnology, Mascheroder Weg 1, 38124-Braunschweig, Germany⁵

Received 15 February 2005/ Accepted 14 April 2005

The Liverpool epidemic strain (LES) of Pseudomonas aeruginosa is a transmissible aggressive pathogen of cystic fibrosis (CF) patients. We compared transcriptome profiles of two LES isolates with each other and with a laboratory and genetic reference strain (PAO1) after growth to late exponential phase and following exposure to oxidative stress. Both LES isolates exhibited enhanced antimicrobial resistances linked to specific mutations in efflux pump genes. Although transcription of AmpC ß-lactamase was up-regulated in both, one LES isolate contained a specific mutation rendering the ampC gene untranslatable. The virulence-related quorum-sensing (QS) regulon of LES431, an isolate that caused pneumonia in the non-CF parent of a CF patient, was considerably up-regulated in comparison to either isolate LES400, associated with a chronic CF infection, or strain PAO1. Premature activation of QS genes was detected in isolates from both non-CF parents and the CF patient in a previously reported infection episode. LES isolates lacking the up-regulated QS phenotype contained different frameshift mutations in lasR. When fed to Drosophila melanogaster, isolate LES431 killed the fruit flies more readily than either isolate LES400 or strain PAO1, indicating that virulence varies intraclonally. The LES may represent a clone with enhanced virulence and antimicrobial resistance characteristics that can vary or are lost due to mutations during long-term colonization but have contributed to the successful spread of the lineage throughout the CF population of the United Kingdom.

Present address: Department of Microbial Pathogenesis and Vaccine Research, German Research Centre for Biotechnology, Mascheroder Weg 1, 38124 Braunschweig, Germany.

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