Correlation of quinolone-resistance, qnr genes and integron carriage in multidrug-resistant community isolates of Klebsiella spp.

Document Type : Original Article

Authors

1 Department of Microbiology, Faculty of Life Sciences and Technology, Shahid Beheshti University, Tehran, Iran

2 Department of Microbiology, Research Center for Food Hygiene and Safety, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

Abstract

Objective(s): Plasmid-mediated quinolone resistance (PMQR) determinants and integrons have a considerable contribution to bacterial drug resistance in Gram-negative pathogens. We studied the prevalence of PMQR genes and integron carriage in multidrug-resistant community isolates of Klebsiella spp.
Materials and Methods: Two hundred and fifty Klebsiella spp. isolates were collected from outpatient specimens between August 2015 and October 2017 in Yazd central Laboratory, Iran. Antibiotic susceptibility was determined against 17 antibiotics and minimum inhibitory concentration (MIC) of ciprofloxacin was measured by E-test. Polymerase chain reaction (PCR) was employed for detection of qnrA, qnrB, qnrS, aac(6’)-Ib-cr, oqxAB and qepA genes.
Results: Disc diffusion results showed that 17 isolates (6.8%) were multidrug resistant (MDR), two of which were Klebsiella oxytoca and 15 were Klebsiella pneumoniae. MIC measurements revealed 11 ciprofloxacin-resistant isolates (including the two K. oxytoca), three intermediately-resistant and three ciprofloxacin-susceptible isolates. All ciprofloxacin-resistant and intermediately-resistant isolates carried at least one and up to four PMQR genes. The most prevalent PMQR gene was oqxAB (93.75%) followed by aac(6’)-ib-cr (50.0%), qnrB (25.0%) and qnrS (18.75%) but qnrA and qepA were not detected. Class 1 integron was observed in 14 (82.3%) isolates including nine ciprofloxacin-resistant, two intermediately-resistant, and three susceptible isolates. Class 2 and 3 integrons were not observed.
Conclusion: Presence of MDR, multiple PMQR determinants as well as class 1 integron in community isolates of Klebsiella spp. can be an important source of transmission of these opportunistic pathogens.

Keywords


1. Paczosa MK, Mecsasb J. Klebsiella pneumoniae: Going on the offense with a strong defense. Microbiol Mol Biol Rev 2016; 80:629-661.
2. Jadhav S, Misra R, Gandham N, Ujagare M, a Ghosh P, Angadiand K, et al. Increasing incidence of multidrug resistant Klebsiella pneumoniae infections in hospital and community settings. Inter J Microbiol Res 2012; 4:253-257.
3.  Seyedpour SM, Eftekhar F. Quinolone susceptibility and detection of qnr and aac(6’)-Ib-cr genes in community isolates of Klebsiella pneumoniae. Jundishapur J Microbiol 2014; 7:1-4.
4. Hooper DC. Mechanisms of quinolone resistance. Drug Resist updates 2003; 2:38-55.
5. Jacoby GA, Strahilevitz J,  Hooper DC. Plasmid-mediated quinolone resistance. Microbiol Spectrum 2014; 2:1-24.
6. Raei F, Eftekhar F, Feizabadi MM. Prevalence of quinolone resistance among extended-spectrum β-lactamase producing uropathogenic Klebsiella pneumoniae. Jundishapur J Microbiol 2014; 7:e10887.
7. Briales A, Rodríguez-Martínez JM, Velasco C, de Alba PD, Rodríguez-Baño J, Martínez-Martínez L, et al. Prevalence of plasmid-mediated quinolone resistance determinants qnr and aac(6’)-Ib-cr in Escherichia coli and Klebsiella pneumoniae producing extended-spectrum.  Int J Antimicrob Agents 2012; 39:431-434.
8. Rahman Z, Islam A, Rashid MU, Johura FT, Monira S, Watanabe H, et al. Existence of a novel qepA variant in quinolone resistant Escherichia coli from aquatic habitats of Bangladesh. Gut Pathol 2017; 9:58-61.
9. Pakzad I, Karin MZ, Taherikalani M, Boustanshenas M, Lari AR. Contribution of AcrAB efflux pump to ciprofloxacin resistance in Klebsiella pneumoniae isolated from burn patients. GMS Hyg Infec Cont 2013; 8:1-6.
10. Rodríguez-Martínez JM, Díaz de Alba P, Briales A, Machuca J, Lossa M, Fernández-Cuenca F, et al. Contribution of OqxAB efflux pumps to quinolone resistance in extended-spectrum-β-lactamase-producing Klebsiella pneumoniae. J Antimicrob Chemother 2013; 68:68-73.
11. Sorensen AH, Hansen LH, Johannesen E, Sorensen SJ. Conjugative plasmid conferring resistance to olaquindox. Antimicrob Agents Chemother 2003; 47:798-799.
12. Park KS, Kim MH, Park TS, Nam YS, Lee HJ, Suh JT. Prevalence of the plasmid-mediated quinolone resistance genes, aac(6’)-Ib-cr, qepA, and oqxAB in clinical isolates of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae in Korea. Ann Clin Lab Sci 2012; 42:191-197.  
13. Yuan J, Xu X, Guo Q, Zhao X, Ye X, Guo Y, et al. Prevalence of the oqxAB gene complex in Klebsiella pneumoniae and Escherichia coli clinical isolates. J Antimicrob Chemother 2012; 67:1655-1659.
14. Navon-Venezia S, Kondratyeva K, Carattoli A. Klebsiella pneumoniae: a major worldwide source and shuttle for antibiotic resistance. FEMS Microbiol Rev 2017; 41:252-275.
15. Wen Y, Pu X, Zheng W, Hu G. High prevalence of plasmid-mediated quinolone resistance and IncQ plasmids carrying qnrS2 gene in bacteria from rivers near hospitals and aquaculture in China. PLoS ONE 2016; 11:e0159418.
16. Akya A, Elahi A, Chegenelorestani R, Rezaee M. Dissemination of multidrug-resistant, class I and II integrons and molecular typing of CTX-M-producing Klebsiella pneumoniae. Int’l J Appl Basic Med Res 2018; 8:100-105.
17. Deng Y, Bao X, Ji L, Chen L, Liu J, Miao J, et al. Resistance integrons: class 1, 2 and 3 integrons. Ann Clin Microbiol Antimicrob 2015; 14:45-55.
18. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. 27th Informational Supplement. M100-S17, Clinical and Laboratory Standards Institute, Wayne, PA: 2017.
19. Chen X, Zhang W, Pan W, Yin J, Pan Z, Gao S, et al. Prevalence of qnr, aac(6’)-Ib-cr, qepA, and oqxAB in Escherichia coli isolates from humans, animals, and the environment. Antimicrob Agents Chemother 2012; 56:3423-3427.
20. Khoramrooz SS, Sharifi A, Yazdanpanah M, Malek Hosseini SA, Emaneini M, Gharibpour F, et al. High frequency of class 1 integrons in Escherichia coli isolated from patients with urinary tract infections in Yasuj, Iran. Iran Red Crescent Med J 2016; 18:e26399.
21. Hashemi SH, Esna-Ashari F, Tavakoli S, Mamani M. The prevalence of antibiotic resistance of Enterobacteriaceae strains isolated in community and hospital-acquired infections in teaching hospitals of Hamadan, West of Iran, J Res Health Sci 2013; 13:75-80.
22. Barguigua A, EI Otmani F, Talmi M, Bourjilat F, Haouzane F, Zerouali K, et al. Characterization of extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumoniae isolates from the community in Morocco. J Med Microbiol 2011; 60:1344-1352.
23. Barguigua A, EI Otmani F, Talmi M, Reguig A, Jamali L, Zerouali K, et al. Prevalence and genotypic analysis of plasmid-mediated beta-lactamases among urinary Klebsiella pneumoniae isolates in Moroccan community. J Antibiotics 2013; 66:11-16.
24. Le TM, Baker S, Le TP, Le TP, Cao TT, Tran TT, et al. High prevalence of plasmid-mediated quinolone resistance determinants in commensal members of the Enterobacteriaceae in Ho Chi Minh City, Vietnam. J Med Microbiol 2009; 58:1585-1592.
25. Dakic I, Lilakos D, Svabic-Vlahovic M, Christofilopoulou S, Poggas N, Charvalos E. Integron-associated antimicrobial resistance in isolates of members of Enterobacteriaceae causing community-acquired urinary infections. Microb Ecol Health Dis 2007; 19:201-206.
26. Araújo BF​,  de Campos PA​,  Royer S​,  Ferreira ML,  Gonçalves IR,  da Fonseca Batistão DW. High frequency of the combined presence of QRDR mutations and PMQR determinants in multidrug-resistant Klebsiella pneumoniae and Escherichia coli isolates from nosocomial and community-acquired infections. J Med Microbiol 2017; 66:1144-1150.
27. Leverstein-van Hall MA, Paauw A, Box AT, Blok HE, Verhoef J Fluit AC. Presence of integron associated resistance in the community is widespread and contributes to multidrug resistance in the hospital. J Clin Microbiol 2002; 40:3038-3040.
28. Kaplan E, Ofek M, Jurkevitch E, Cytryn E. Characterization of fluoroquinolone resistance and qnr diversity in Enterobacteriaceae from municipal biosolids. Front Microbiol 2013; 4:144-150.