Infect Chemother.  2015 Jun;47(2):98-104. 10.3947/ic.2015.47.2.98.

Spread of Efflux Pump Overexpressing-Mediated Fluoroquinolone Resistance and Multidrug Resistance in Pseudomonas aeruginosa by using an Efflux Pump Inhibitor

• Adabi M
• Talebi-Taher M
• Arbabi L
• Afshar M
• Fathizadeh S
• Minaeian S
• Moghadam-Maragheh N
• Majidpour

• KMID: 2068993
• DOI: http://doi.org/10.3947/ic.2015.47.2.98

Abstract

BACKGROUND
Fluoroquinolone resistance in Pseudomonas aeruginosa may be due to efflux pump overexpression and/or target mutations. We designed this study to investigate the efflux pump mediated fluoroquinolone resistance and check the increasing effectiveness of fluoroquinolones in combination with an efflux pumps inhibitor among P. aeruginosa isolates from burn wounds infections.
MATERIALS AND METHODS
A total of 154 consecutive strains of P. aeruginosa were recovered from separate patients hospitalized in a burn hospital, Tehran, Iran. The isolates first were studied by disk diffusion antibiogram for 11 antibiotics and then minimum inhibitory concentration (MIC) experiments were performed to detect synergy between ciprofloxacin and the efflux pump inhibitor, carbonyl cyanide-m-chlorophenyl hydrazone (CCCP). Then to elucidate the inducing of multi drug resistance due to different efflux pumps activation in Fluoroquinolone resistant isolates, synergy experiments were also performed in random ciprofloxacin resistant isolates which have overexpressed efflux pumps phenotypically, using CCCP and selected antibiotics as markers for Beta-lactams and Aminoglycosides. The isolates were also tested by polymerase chain reaction (PCR) for the presence of the MexA, MexC and MexE, which encode the efflux pumps MexAB-OprM, MexCD-OprJ and MexEF-OprN.
RESULTS
Most of the isolates were resistant to 3 or more antibiotics tested. More than half of the ciprofloxacin resistant isolates exhibited synergy between ciprofloxacin and CCCP, indicating the efflux pump activity contributed to the ciprofloxacin resistance. Also increased susceptibility of random ciprofloxacin resistant isolates of P. aeruginosa to other selected antibiotics, in presence of CCCP, implied multidrug extrusion by different active efflux pump in fluoroquinolones resistant strains. All of Ciprofloxacin resistant isolates were positive for MexA, MexC and MexE genes simultaneously.
CONCLUSION
In this burn hospital, where multidrug resistant P. aeruginosa isolates were prevalent, ciprofloxacin resistance and multidrug resistance due to the overexpression of fluoroquinolones mediated efflux pumps has also now emerged. Early recognition of this resistance mechanism should allow the use of alternative antibiotics and use an efflux pumps inhibitor in combination with antibiotic therapy.

Keyword

Pseudomonas aeruginosa; Antibiotic therapy; Efflux pump; Ciprofloxacin, Carbonyl cyanide-m-chlorophenyl hydrazone; Burned patients

MeSH Terms

Aminoglycosides
Anti-Bacterial Agents
beta-Lactams
Burns
Carbonyl Cyanide m-Chlorophenyl Hydrazone
Ciprofloxacin
Diffusion
Drug Resistance
Drug Resistance, Multiple*
Fluoroquinolones
Humans
Iran
Microbial Sensitivity Tests
Polymerase Chain Reaction
Pseudomonas aeruginosa*
Wounds and Injuries
Aminoglycosides
Anti-Bacterial Agents
Carbonyl Cyanide m-Chlorophenyl Hydrazone
Ciprofloxacin
Fluoroquinolones
beta-Lactams

Figure

• 

  Figure 1 The susceptibility status of the isolates against 11 studied antibiotics according to disk diffusion. IMI, imipenem; CEF, cefepime; TC, ticarcillin; AT, aztreonam; TOB, tobramycin; GN, gentamicin; CO, colistin; CIP, ciprofloxacin; AK, amikacin; PTZ, piperacillin-tazobactam; PIP, piperacillin.

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