CC BY-NC-ND 4.0 · J Lab Physicians 2022; 14(03): 271-277
DOI: 10.1055/s-0042-1742636
Original Article

Plasmid-Mediated Fluoroquinolone Resistance in Pseudomonas aeruginosa and Acinetobacter baumannii

Geetha P. Venkataramana
1   Department of Microbiology, Sri Ramachandra Institute of Higher Education and Research, Porur, Chennai, Tamil Nadu, India
,
Aishwarya K.V. Lalitha
1   Department of Microbiology, Sri Ramachandra Institute of Higher Education and Research, Porur, Chennai, Tamil Nadu, India
,
Shanthi Mariappan
1   Department of Microbiology, Sri Ramachandra Institute of Higher Education and Research, Porur, Chennai, Tamil Nadu, India
,
Uma Sekar
1   Department of Microbiology, Sri Ramachandra Institute of Higher Education and Research, Porur, Chennai, Tamil Nadu, India
› Author Affiliations
Funding This study is funded by Department of Science & Technology (DST) under Women Scientists Scheme (WOS-A).

Abstract

IntroductionPseudomonas aeruginosa and Acinetobacter baumannii are important pathogens in health care–associated infections. Fluoroquinolone resistance has emerged in these pathogens. In this study, we aimed to determine the occurrence of plasmid-mediated quinolone resistance (PMQR) determinants (qnrA, qnrB, qnrS, aac(6′)-Ib-cr, oqxAB, and qepA) by polymerase chain reaction (PCR) and the transmissibility of plasmid-borne resistance determinants in clinical isolates of P. aeruginosa and A. baumannii.

Materials and Methods The study included P. aeruginosa (85) and A. baumannii (45) which were nonduplicate, clinically significant, and ciprofloxacin resistant. Antibiotic susceptibility testing was done by disk diffusion method for other antimicrobial agents, namely amikacin, ceftazidime, piperacillin/tazobactam, ofloxacin, levofloxacin, and imipenem. Minimum inhibitory concentration of ciprofloxacin was determined. Efflux pump activity was evaluated using carbonyl-cyanide m-chlorophenylhydrazone (CCCP). The presence of PMQR genes was screened by PCR amplification. Transferability of PMQR genes was determined by conjugation experiment, and plasmid-based replicon typing was performed.

Results Resistance to other classes of antimicrobial agents was as follows: ceftazidime (86.9%), piperacillin/tazobactam (73.8%), imipenem (69.2%), and amikacin (63.8%). The minimal inhibitory concentration (MIC)50 and MIC90 for ciprofloxacin were 64 and greater than or equal to 256 µg/mL, respectively. There was a reduction in MIC for 37 (28.4%) isolates with CCCP. In P. aeruginosa, 12 (14.1%) isolates harbored qnrB, 12 (14.1%) qnrS, 9 (10.5%) both qnrB and qnrS, 66 (77.6%) aac(6′)-Ib-cr, and 3 (3.5%) oqxAB gene. In A. baumannii, qnrB was detected in 2 (4.4%), 1 (2.2%) harbored both the qnrA and qnrS, 1 isolate harbored qnrB and qnrS, 21 (46.6%) aac(6′)-Ib-cr, and 1 (2.2%) isolate harbored oqxAB gene. Notably, qepA gene was not detected in any of the study isolates. Conjugation experiments revealed that 12 (9.2%) were transferable. Of the transconjugants, seven (58.3%) belonged to IncFII type plasmid replicon, followed by four (33.3%) IncA/C and one (8.3%) IncFIC type.

Conclusion The plasmid-mediated resistance aac(6′)-Ib-cr gene is primarily responsible for mediating fluoroquinolone resistance in clinical isolates of P. aeruginosa and A. baumannii. The predominant plasmid type is IncFII.



Publication History

Article published online:
09 February 2022

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  • References

  • 1 Wang M, Tran JH, Jacoby GA, Zhang Y, Wang F, Hooper DC. Plasmid-mediated quinolone resistance in clinical isolates of Escherichia coli from Shanghai, China. Antimicrob Agents Chemother 2003; 47 (07) 2242-2248
  • 2 Bonomo RA, Szabo D. Mechanisms of multidrug resistance in Acinetobacter species and Pseudomonas aeruginosa . Clin Infect Dis 2006; 43 (2, Suppl 2): S49-S56
  • 3 Hooper DC, Jacoby GA. Mechanisms of drug resistance: quinolone resistance. Ann N Y Acad Sci 2015; 1354 (01) 12-31
  • 4 Martínez-Martínez L, Pascual A, Jacoby GA. Quinolone resistance from a transferable plasmid. Lancet 1998; 351 (9105): 797-799
  • 5 Robicsek A, Jacoby GA, Hooper DC. The worldwide emergence of plasmid-mediated quinolone resistance. Lancet Infect Dis 2006; 6 (10) 629-640
  • 6 Yamane K, Wachino J, Suzuki S. et al. New plasmid-mediated fluoroquinolone efflux pump, QepA, found in an Escherichia coli clinical isolate. Antimicrob Agents Chemother 2007; 51 (09) 3354-3360
  • 7 Jacoby GA, Strahilevitz J, Hooper DC. Plasmid-mediated quinolone resistance. Microbiol Spectr 2014; 2 (05) 10
  • 8 Rodríguez-Martínez JM, Díaz de Alba P, Briales A. et al. Contribution of OqxAB efflux pumps to quinolone resistance in extended-spectrum-β-lactamase-producing Klebsiella pneumoniae . J Antimicrob Chemother 2013; 68 (01) 68-73
  • 9 Rodríguez-Martínez JM, Cano ME, Velasco C, Martínez-Martínez L, Pascual A. Plasmid-mediated quinolone resistance: an update. J Infect Chemother 2011; 17 (02) 149-182
  • 10 Yang H, Chen H, Yang Q, Chen M, Wang H. High prevalence of plasmid-mediated quinolone resistance genes qnr and aac(6′)-Ib-cr in clinical isolates of Enterobacteriaceae from nine teaching hospitals in China. Antimicrob Agents Chemother 2008; 52 (12) 4268-4273
  • 11 Zhu YL, Yang H-F, Liu YY. et al. Detection of plasmid-mediated quinolone resistance determinants and the emergence of multidrug resistance in clinical isolates of Shigella in SiXian area, China. Diagn Microbiol Infect Dis 2013; 75 (03) 327-329
  • 12 Yang H, Hu L, Liu Y, Ye Y, Li J. Detection of the plasmid-mediated quinolone resistance determinants in clinical isolates of Acinetobacter baumannii in China. J Chemother 2016; 28 (05) 443-445
  • 13 Carattoli A, Bertini A, Villa L, Falbo V, Hopkins KL, Threlfall EJ. Identification of plasmids by PCR-based replicon typing. J Microbiol Methods 2005; 63 (03) 219-228
  • 14 Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Susceptibility Testing. M100. 27th ed.. Wanye (PA): Clinical and Laboratory Standards Institute; 2017
  • 15 Ardebili A, Talebi M, Azimi L, Rastegar Lari A. Effect of efflux pump inhibitor carbonyl cyanide 3-chlorophenylhydrazone on the minimum inhibitory concentration of ciprofloxacin in Acinetobacter baumannii clinical isolates. Jundishapur J Microbiol 2014; 7 (01) e8691
  • 16 Robicsek A, Strahilevitz J, Sahm DF, Jacoby GA, Hooper DC. qnr prevalence in ceftazidime-resistant Enterobacteriaceae isolates from the United States. Antimicrob Agents Chemother 2006; 50 (08) 2872-2874
  • 17 Wareham DW, Umoren I, Khanna P, Gordon NC. Allele-specific polymerase chain reaction (PCR) for rapid detection of the aac(6′)-Ib-cr quinolone resistance gene. Int J Antimicrob Agents 2010; 36 (05) 476-477
  • 18 Saleh MA, Balboula MM. Plasmid mediated quinolone resistance determinants among nosocomial clinical Pseudomonas aeruginosa isolates. Int J Curr Microbiol Appl Sci 2017; 6 (01) 42-50
  • 19 Wang M, Tran JH, Jacoby GA, Zhang Y, Wang F, Hooper DC. Plasmid-mediated quinolone resistance in clinical isolates of Escherichia coli from Shanghai, China. Antimicrob Agents Chemother 2003; 47 (07) 2242-2248
  • 20 Pham TDM, Ziora ZM, Blaskovich MAT. Quinolone antibiotics. MedChemComm 2019; 10 (10) 1719-1739
  • 21 Navon-Venezia S, Ben-Ami R, Carmeli Y. Update on Pseudomonas aeruginosa and Acinetobacter baumannii infections in the healthcare setting. Curr Opin Infect Dis 2005; 18 (04) 306-313
  • 22 Zaki MES, Abou ElKheir N, Mofreh M. Molecular study of quinolone resistance determining regions of gyrA gene and parC genes in clinical isolates of Acintobacter baumannii resistant to fluoroquinolone. Open Microbiol J 2018; 12: 116-122
  • 23 Chenia HY, Pillay B, Pillay D. Analysis of the mechanisms of fluoroquinolone resistance in urinary tract pathogens. J Antimicrob Chemother 2006; 58 (06) 1274-1278
  • 24 Osei Sekyere J, Amoako DG. Genomic and phenotypic characterisation of fluoroquinolone resistance mechanisms in Enterobacteriaceae in Durban, South Africa. PLoS One 2017; 12 (06) e0178888
  • 25 Nikaido H, Pagès JM. Broad-specificity efflux pumps and their role in multidrug resistance of Gram-negative bacteria. FEMS Microbiol Rev 2012; 36 (02) 340-363
  • 26 Talebi-Taher M, Majidpour A, Gholami A, Rasouli-Kouhi S, Adabi M. Role of efflux pump inhibitor in decreasing antibiotic cross-resistance of Pseudomonas aeruginosa in a burn hospital in Iran. J Infect Dev Ctries 2016; 10 (06) 600-604
  • 27 Al Rashed N, Joji RM, Saeed NK, Bindayna KM. Detection of overexpression of efflux pump expression in fluoroquinolone-resistant Pseudomonas aeruginosa isolates. Int J Appl Basic Med Res 2020; 10 (01) 37-42
  • 28 Helmy OM, Kashef MT. Different phenotypic and molecular mechanisms associated with multidrug resistance in Gram-negative clinical isolates from Egypt. Infect Drug Resist 2017; 10: 479-498
  • 29 Gomaa FM, Tawakol WM, El-Azm FI. Phenotypic and genotypic detection of some antimicrobial resistance mechanisms among multidrug-resistant Acinetobacter baumannii isolated from immunocompromised patients in Egypt. Egypt J Med Microbiol 2014; 23 (04) 99-111
  • 30 El-Badawy MF, Alrobaian MM, Shohayeb MM, Abdelwahab SF. Investigation of six plasmid-mediated quinolone resistance genes among clinical isolates of pseudomonas: a genotypic study in Saudi Arabia. Infect Drug Resist 2019; 12: 915-923
  • 31 Rafiq K, Ahmad K, Ahmad N, Gohar M, Shehzad MA, Saeed MQ. Determination of Qnr allele frequencies in fluoroquinolone resistant Pseudomonas aeruginosa isolated from burn wounds. J Pak Med Assoc 2019; 69 (02) 250-252
  • 32 Yang X, Xing B, Liang C, Ye Z, Zhang Y. Prevalence and fluoroquinolone resistance of Pseudomonas aeruginosa in a hospital of South China. Int J Clin Exp Med 2015; 8 (01) 1386-1390
  • 33 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 (04) 310-312
  • 34 Coban AY, Tanrıverdi Çaycı Y, Yıldırım T, Erturan Z, Durupınar B, Bozdoğan B. Investigation of plasmid-mediated quinolone resistance in Pseudomonas aeruginosa strains isolated from cystic fibrosis patients [in Turkish]. Mikrobiyol Bul 2011; 45 (04) 602-608
  • 35 Hamed SM, Elkhatib WF, El-Mahallawy HA, Helmy MM, Ashour MS, Aboshanab KMA. Multiple mechanisms contributing to ciprofloxacin resistance among Gram negative bacteria causing infections to cancer patients. Sci Rep 2018; 8 (01) 12268
  • 36 Touati A, Brasme L, Benallaoua S, Gharout A, Madoux J, De Champs C. First report of qnrB-producing Enterobacter cloacae and qnrA-producing Acinetobacter baumannii recovered from Algerian hospitals. Diagn Microbiol Infect Dis 2008; 60 (03) 287-290
  • 37 Araujo BF, Ferreira ML, Campos PA. et al. Clinical and molecular epidemiology of multidrug-resistant P. aeruginosa carrying aac(6′)-Ib-cr, qnrS1 and blaSPM genes in Brazil. PLoS One 2016; 11 (05) e0155914
  • 38 Cayci YT, Coban AY, Gunaydin M. Investigation of plasmid-mediated quinolone resistance in Pseudomonas aeruginosa clinical isolates. Indian J Med Microbiol 2014; 32 (03) 285-289
  • 39 Jafari M, Fallah F, Borhan RS. et al. The first report of CMY, aac (6′)-Ib and 16S rRNA methylase genes among Pseudomonas aeruginosa isolates from Iran. Arch Pediatr Infect Dis 2013; 1 (03) 109-112
  • 40 Xue-qing Z, Dan-ping L, Chun-quan X. et al. Detection of plasmid-mediated quinolone resistance determinants in clinical non-fermentative bacteria and ciprofloxacin sensitive Enterobacteriaceae strains. Dis Surveill 2014; 29 (02) 130-135
  • 41 Michalska AD, Sacha PT, Ojdana D, Wieczorek A, Tryniszewska E. Prevalence of resistance to aminoglycosides and fluoroquinolones among Pseudomonas aeruginosa strains in a University Hospital in Northeastern Poland. Braz J Microbiol 2015; 45 (04) 1455-1458
  • 42 Jiang X, Yu T, Jiang X, Zhang W, Zhang L, Ma J. Emergence of plasmid-mediated quinolone resistance genes in clinical isolates of Acinetobacter baumannii and Pseudomonas aeruginosa in Henan, China. Diagn Microbiol Infect Dis 2014; 79 (03) 381-383
  • 43 Ogbolu DO, Daini OA, Ogunledun A, Terry Alli OA, Webber MA. Dissemination of IncF plasmids carrying beta-lactamase genes in Gram-negative bacteria from Nigerian hospitals. J Infect Dev Ctries 2013; 7 (05) 382-390
  • 44 Elena A, Quinteros M, Di Conza J, Gutkind G, Cejas D, Radice MA. Full characterization of an IncR plasmid harboring qnrS1 recovered from a VIM-11-producing Pseudomonas aeruginosa . Rev Argent Microbiol 2020; 52 (04) 298-304
  • 45 Ogbolu DO, Alli AO, Anorue MC, Daini OA, Oluwadun A. Distribution of plasmid-mediated quinolone resistance in Gram-negative bacteria from a tertiary hospital in Nigeria. Indian J Pathol Microbiol 2016; 59 (03) 322-326
  • 46 Castanheira M, Mendes RE, Jones RN. Update on Acinetobacter species: mechanisms of antimicrobial resistance and contemporary in vitro activity of minocycline and other treatment options. Clin Infect Dis 2014; 59 (6, suppl 6): S367-S373