Figen ERSOY, Esra BÜYÜKCANGAZ, Songül SONAL, Pınar ŞAHİNTÜRK, Erdem ARSLAN, Ayşin ŞEN, Murat CENGİZ, A. Mark WEBBER, J V. Laura PIDDOCK

High level fluoroquinolone resistance in Escherichia coli isolatedfrom animals in Turkey is due to multiple mechanisms

The aim of this study was to determine the molecular mechanisms of fluoroquinolone resistance in E. coli isolated from cattle, goats, sheep, cats, and dogs in Turkey. Twenty nonreplicate E. coli isolates (chosen on the basis of RAPD pattern) from food-producing animals were selected for the study. To identify phenotypic differences between isolates, the sum of the MIC values of 14 antimicrobials was calculated; values ranged from 565 to 2520 μg/mL, indicating the diversity in antimicrobial resistance present in the panel of isolates. PCR and qRT-PCR were used to characterize the presence and expression levels of known molecular mechanisms of fluoroquinolone resistance. The number of E. coli isolates having single, double, and triple topoisomerase mutations was 2, 10, and 5, respectively. Moreover, the number of qnrA - , qnrS - , oqxB -, and aac(6 )Ib-cr-containing E. coli isolates was 1, 4, 1, and 17, respectively. Increased expression of acrB and soxS was detected in 2 and 9 isolates, respectively. The results of this study show a wide range of different mechanisms of fluoroquinolone resistance in E. coli isolates in Turkey.

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1. Mavroidi A, Vivi M, Liakopoulos A, Tzelepi E, Stefos A, Dalekos, GN, Petinaki E. Ciprofloxacin-resistant Escherichia coli in Central Greece: mechanisms of resistance and molecular identification. BMC Infect Dis 2012; 12: 371.
2. Hopkins KL, Davies RH, Threlfall EJ. Mechanisms of quinolone resistance in Escherichia coli and Salmonella: recent developments. Int J Antimicrob Ag 2005; 25: 358373.
3. Hansen LH, Jensen LB, Sorensen HI, Sorensen SJ. Substrate specificity of the OqxAB multidrug resistance pump in Escherichia coli and selected enteric bacteria. J Antimicrob Chemother 2007; 60: 145147.
4. Yamane K, Wachino J, Suzuki S, Arakawa Y. Plasmid-mediated qepA gene among Escherichia coli clinical isolates from Japan. Antimicrob Agents Chemother 2008; 52: 15641566.
5. Wang M, Guo O, Xu X, Wang X, Ye X, Wu S, Hooper DC, Wang M. New plasmid-mediated quinolone resistance gene, qnrC, found in a clinical isolate of Proteus mirabilis. Antimicrob Agents Chemother 2009; 53: 18921897.
6. Bambeke FV, Pages JM, Lee VJ. Inhibitors of bacterial efflux pumps as adjuvants in antibiotic treatments and diagnostic tools for detection of resistance by efflux. Recent Pat Antiinfect Drug Discov 2006; 1: 157175.
7. Webber M, Piddock LJV. Quinolone resistance in Escherichia coli. Vet Res 2001; 32: 275284.
8. Nazik H, Ongen B, Kuvat N. Investigation of plasmid mediated quinolone resistance among isolates obtained in a Turkish intensive care unit. Jpn J Infect Dis 2008; 61: 310312.
9. Barbour EK, Ahmadieh D, Harakeh S, Kumosani T. Impact of antimicrobials use in chickens on emergence of drug-resistant Campylobacter organisms in humans. Int J Antimicrob Ag 2012; 2: 1.
10. Clinical Laboratory Standards Institute. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically. 6th ed. Approved Standard M7-A6. 2009; Wayne, PA, USA: CLSI; 2010.
11. The European Committee on Antimicrobial Susceptibility Testing (EUCAST) 2014. Breakpoint tables for interpretation of MICs and zone diameters. Vaxjo, Sweden: EUCAST; 2014.
12. Adelowo OO, Fagade OE, Adelowo AY. Antibiotic resistance and resistance genes in Escherichia coli from poultry farms, southwest Nigeria. J Infect Dev Ctries 2014; 8: 11031112.
13. Robicsek A, Strahilevitz J, Sahm DF, Jacoby GA, Hooper DC. qnr prevalence in ceftazidime-resistant Enterobacteriaceaeisolates from United States. Antimicrob Agents Chemother 2006; 50: 28722874.
14. Perichon B, Courvalin P, Galimand M. Transferable resistance to aminoglycosides by methylation of G1450 in 16S rRNA and to hydrophilic fluoroquinolones by QepA-mediated efflux in Escherichia coli. Antimicrob Agents Chemother 2007; 51: 24642469.
15. Park CH, Robicsek A, Jacoby GA, Sahm D, Hooper DC. Prevalence in the United States of aac(6)Ib-cr encoding a ciprofloxacin-modifying enzyme. Antimicrob Agents Chemother 2006; 50: 39533955.
16. Karczmarczyk M, Martins M, Quinn T, Leonard N, Fanning S. Mechanisms of fluoroquinolone resistance in Escherichia coliisolates from food-producing animals. App Environ Microbiol 2011; 7: 71137120.
17. Coldham NG, Webber MA, Woodward MJ, Piddock LJ. A 96-well plate fluorescence assay for assessment of cellular permeability and active efflux in Salmonella enterica serovar Typhimurium and Escherichia coli. J Antimicrob Chemother 2010; 65: 165563.
18. Afifi MM, Suelam IIA, Soliman MTA, El-Gohary MGS. Prevalence and antimicrobial susceptibility pattern of Pseudomonas aeruginosa isolated from environmental and clinical samples in upper Egypt. IJBCS 2013; 7: 4757.
19. Wong MH, Chan EW, Liu LZ, Chen S. PMQR genes oqxABand aac(6)Ib-cr accelerate the development of fluoroquinolone resistance in Salmonella typhimurium. Front Microbiol 2014; 5: 521.
20. Robicsek A, Jacoby G, Hooper DC. The worldwide emergence of plasmid-mediated quinolone resistance. Lancet Infect Dis 2006; 6: 629640.
21. Martinez-Martinez L, Pascual A, Jacoby G. Quinolone resistance from a transferable plasmid. Lancet 1998; 351: 797799.
22. ORegan E, Quinn T, Pages JM, Mccusker M, Piddock L, Fanning S. Multiple regulatory pathways associated with high-level ciprofloxacin and multidrug resistance in Salmonella enterica serovar enteritidis: involvement of ramA and other global regulators. Antimicrob Agents Chemother 2008; 53: 10801087.
23. Piddock LJV. Understanding the basis of antibiotic resistance: a platform for drug discovery. Microbiology 2014; 160: 23662373.
24. Everett MJ, Jin YF, Ricci V, Piddock LJV. Contributions of individual mechanisms to fluoroquinolone resistance in 36 Escherichia coli strains isolated from humans and animals. Antimicrob Agents Ch 1996; 40: 23802386.