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JAC Advance Access originally published online on October 4, 2009
Journal of Antimicrobial Chemotherapy 2009 64(6):1212-1218; doi:10.1093/jac/dkp360
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© The Author 2009. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Original research

Restricting ciprofloxacin-induced resistant variant formation in biofilm of Klebsiella pneumoniae B5055 by complementary bacteriophage treatment

Vivek Verma, Kusum Harjai and Sanjay Chhibber*

Department of Microbiology, Basic Medical Sciences (BMS) Block, Panjab University, Chandigarh 160014, India

Received 9 July 2009; returned 12 August 2009; revised 8 September 2009; accepted 9 September 2009

* Corresponding author. Tel: +91-172-2534141/+91-977-9817768; Fax: +91-172-2541770; E-mail: sanjaychhibber8{at}gmail.com

Objectives: The in vitro efficacy of lytic bacteriophage as a therapeutic agent against Klebsiella pneumoniae biofilm, alone or in combination with ciprofloxacin, was studied. The pathogenic potential of the resistant variants formed during the treatment was evaluated.

Methods: A lytic bacteriophage in combination with ciprofloxacin was used for the treatment of K. pneumoniae B5055 biofilm. The efficacy and the frequency of resistant variant formation were estimated after respective treatments. The resistant variants were characterized for their virulence potential.

Results: Bacteriophage alone was able to eradicate the biofilm effectively and no significant difference was observed in its ability to eradicate biofilm in combination with ciprofloxacin. However, combination treatment using ciprofloxacin and bacteriophage significantly arrested the emergence of resistant variants. The small number of variants that developed had a lower propensity to form biofilms, produced small amounts of cell-associated capsular polysaccharide and demonstrated increased susceptibility to mouse peritoneal macrophages. Altered morphology and changed pattern of the outer membrane proteins of bacterial isolates were also observed.

Conclusions: The combination treatment not only killed the bacteria, but also restricted the formation of resistant variants significantly as compared with individual treatments. Hence, a combination of bacteriophage and ciprofloxacin offers an effective strategy to combat the emergence of treatment-associated resistance.

Keywords: K. pneumoniae , resistance , biofilm formation


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