Evaluation of antimicrobial resistance, biofilm forming potential, and the presence of biofilm-related genes among clinical isolates of Pseudomonas aeruginosa

Esmat Kamali, Ailar Jamali, Abdollah Ardebili, Freshteh Ezadi, Alireza Mohebbi, Esmat Kamali, Ailar Jamali, Abdollah Ardebili, Freshteh Ezadi, Alireza Mohebbi

Abstract

Objectives: Pseudomonas aeruginosa is known as a leading cause of nosocomial infections worldwide. Antimicrobial resistance and biofilm production, as two main virulence factors of P. aeruginosa, are responsible for the persistence of prolonged infections. In this study, antimicrobial susceptibility pattern and phenotypic and genotypic characteristics of biofilm of P. aeruginosa were investigated.

Results: A total of 80 clinical P. aeruginosa isolates were obtained. Isolates showed resistance to all antibiotics with a rate from 12.5% (n = 10) against amikacin and piperacillin/tazobactam to 23.75% (n = 19) to levofloxacin. Multidrug-resistant P. aeruginosa accounted for 20% (n = 16). 83.75% (n = 67) of isolates showed biofilm phenotype. All three biofilm-related genes were found simultaneously in 87.5% (n = 70) of P. aeruginosa and 13.5% (n = 10) of the isolates had none of the genes tested. From the results of the present study, combination therapy including an anti-pseudomonal beta-lactam (piperacillin/tazobactam or ceftazidime) and an aminoglycoside or carbapenems (imipenem, meropenem) with fluoroquinolones in conjunction with an aminoglycoside can be used against Pseudomonas infections. However, reasonable antimicrobial use and high standards of infection prevention and control are essential to prevent further development of antimicrobial resistance. Combination strategies based on the proper anti-pseudomonal antibiotics along with anti-biofilm agents can also be selected to eradicate biofilm-associated infections.

Keywords: Antimicrobial resistance; Biofilm formation; Biofilm genes; Pseudomonas aeruginosa.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Antibiotic susceptibility patterns of P. aeruginosa clinical isolates. IMI imipenem, MEM meropenem, GM gentamicin, TOB tobramycin, AK amikacin, CIP ciprofloxacin, LEV levofloxacin, CAZ ceftazidime, PTZ piperacillin/tazobactam
Fig. 2
Fig. 2
PCR amplification of biofilm-encoding genes in one selected clinical isolate of P. aeruginosa as representative. a Lane 1–3: PCR products of the pelF, algD, and pslD genes, respectively. M: 50 bp DNA ladder. Lane 4: PCR mixture without DNA template as control negative. Lane 5–7: PCR products of the corresponding genes in P. aeruginosa PAO1 reference strain as control positive. b A 50 bp DNA ladder containing seventeen discrete fragments ranging from 50 to 1500 bp with double intensity reference bands at 200 bp, 500 bp, and 1200 bp

References

    1. Pachori P, Gothalwal R, Gandhi P. Emergence of antibiotic resistance Pseudomonas aeruginosa in intensive care unit; a critical review. Genes Dis. 2019;6(2):109–119.
    1. Dogonchi AA, Ghaemi EA, Ardebili A, Yazdansetad S, Pournajaf A. Metallo-β-lactamase-mediated resistance among clinical carbapenem-resistant Pseudomonas aeruginosa isolates in northern Iran: a potential threat to clinical therapeutics. Tzu Chi Med J. 2018;30(2):90–96.
    1. Bijari A, Azimi L, Fallah F, Ardebili A, Rastegar Lari E, Rastegar Lari A. Involvement of the multidrug efflux pumps in betalactams resistant Pseudomonas aerugionsa clinical isolates collected from burn patients in Iran. Infect Disord Drug Targets. 2016;16(3):172–177.
    1. Moreira MR, Guimarães MP, Rodrigues AA, Gontijo Filho PP. Antimicrobial use, incidence, etiology and resistance patterns in bacteria causing ventilator-associated pneumonia in a clinical-surgical intensive care unit. Rev Soc Bras Med Trop. 2013;46(1):39–44.
    1. Baker P, Hill PJ, Snarr BD, Alnabelseya N, Pestrak MJ, Lee MJ, et al. Exopolysaccharide biosynthetic glycoside hydrolases can be utilized to disrupt and prevent Pseudomonas aeruginosa biofilms. Sci Adv. 2016;2(5):e1501632.
    1. Franklin MJ, Nivens DE, Weadge JT, Howell PL. Biosynthesis of the Pseudomonas aeruginosa extracellular polysaccharides, alginate, Pel, and Psl. Front Microbiol. 2011;2:167.
    1. Govan JR, Deretic V. Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia. Microbiol Mol Biol Rev. 1996;60(3):539–574.
    1. Banar M, Emaneini M, Satarzadeh M, Abdellahi N, Beigverdi R, van Leeuwen WB, et al. Evaluation of mannosidase and trypsin enzymes effects on biofilm production of Pseudomonas aeruginosa isolated from burn wound infections. PLoS ONE. 2016;11(10):e0164622.
    1. Laverty G, Gorman SP, Gilmore BF. Biomolecular mechanisms of Pseudomonas aeruginosa and Escherichia coli biofilm formation. Pathogens. 2014;3(3):596–632.
    1. Ma L, Conover M, Lu H, Parsek MR, Bayles K, Wozniak DJ. Assembly and development of the Pseudomonas aeruginosa biofilm matrix. PLoS Pathog. 2009;5(3):e1000354.
    1. Campisano A, Schroeder C, Schemionek M, Overhage J, Rehm BH. PslD is a secreted protein required for biofilm formation by Pseudomonas aeruginosa. Appl Environ Microbiol. 2006;72(4):3066–3068.
    1. Ghafoor A, Jordens Z, Rehm BH. Role of PelF in Pel polysaccharide biosynthesis in Pseudomonas aeruginosa. Appl Environ Microbiol. 2013;79(9):2968–2978.
    1. Jennings LK, Storek KM, Ledvina HE, Coulon C, Marmont LS, Sadovskaya I, et al. Pel is a cationic exopolysaccharide that cross-links extracellular DNA in the Pseudomonas aeruginosa biofilm matrix. Proc Natl Acad Sci U S A. 2015;112(36):11353–11358.
    1. Friedman L, Kolter R. Genes involved in matrix formation in Pseudomonas aeruginosa PA14 biofilms. Mol Microbiol. 2004;51(3):675–690.
    1. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. 28th ed. M100. Wayne: Clinical and Laboratory Standards Institute (CLSI); 2018. p. 38.
    1. Magiorakos AP, Srinivasan A, Carey R, Carmeli Y, Falagas M, Giske C, et al. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect. 2012;18(3):268–281.
    1. Stepanović S, Vuković D, Hola V, Bonaventura GD, Djukić S, Ćirković I, et al. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS. 2007;115(8):891–899.
    1. Gill J, Arora S, Khanna S, Kumar KH. Prevalence of multidrug-resistant, extensively drug-resistant, and pandrug-resistant Pseudomonas aeruginosa from a tertiary level intensive care unit. J Glob Infect Dis. 2016;8(4):155–159.
    1. Tümmler B. Emerging therapies against infections with Pseudomonas aeruginosa. F1000Res. 2019
    1. European Centre for Disease Prevention and Control. Surveillance of Antimicrobial Resistance in Europe 2017. . Accessed 15 Nov 2018.
    1. Vaez H, Salehi-Abargouei A, Ghalehnoo ZR, Khademi F. Multidrug resistant Pseudomonas aeruginosa in Iran: a systematic review and metaanalysis. J Glob Infect Dis. 2018;10(4):212–217.
    1. Bavasheh N, Karmostaji A. Antibiotic resistance pattern and evaluation of blaOXA-10, blaPER-1, blaVEB, blaSHV genes in clinical isolates of Pseudomonas aeruginosa isolated from hospital in south of Iran in 2014–2015. Infect Epidemiol Microbiol. 2017;3(1):1–5.
    1. Nikokar I, Tishayar A, Flakiyan Z, Alijani K, Rehana-Banisaeed S, Hossinpour M, et al. Antibiotic resistance and frequency of class 1 integrons among Pseudomonas aeruginosa, isolated from burn patients in Guilan, Iran. Iran J Microbiol. 2013;5(1):36–41.
    1. Faghri J, Nouri S, Jalalifar S, Zalipoor M, Halaji M. Investigation of antimicrobial susceptibility, class I and II integrons among Pseudomonas aeruginosa isolates from hospitalized patients in Isfahan, Iran. BMC Res Notes. 2018;11(1):806.
    1. Pournajaf A, Razavi S, Irajian G, Ardebili A, Erfani Y, Solgi S, et al. Integron types, antimicrobial resistance genes, virulence gene profile, alginate production and biofilm formation in Iranian cystic fibrosis Pseudomonas aeruginosa isolates. Infez Med. 2018;26(3):226–236.
    1. Vasiljević Z, Jovčić B, Ćirković I, Đukić S. An examination of potential differences in biofilm production among different genotypes of Pseudomonas aeruginosa. Arch Biol Sci. 2014;66(1):117–121.
    1. Ghadaksaz A, Fooladi AAI, Hosseini HM, Amin M. The prevalence of some Pseudomonas virulence genes related to biofilm formation and alginate production among clinical isolates. J App Biomed. 2015;13(1):61–68.
    1. Müsken M, Di Fiore S, Dötsch A, Fischer R, Häussler S. Genetic determinants of Pseudomonas aeruginosa biofilm establishment. Microbiol. 2010;156(2):431–441.
    1. Moradali MF, Ghods S, Rehm BH. Pseudomonas aeruginosa lifestyle: a paradigm for adaptation, survival, and persistence. Front Cell Infect Microbiol. 2017;7:39.
    1. Hou W, Sun X, Wang Z, Zhang Y. Biofilm-forming capacity of Staphylococcus epidermidis, Staphylococcus aureus, and Pseudomonas aeruginosa from ocular infections. Invest Ophthalmol Vis Sci. 2012;53(9):5624–5631.
    1. Lima JLdC, Alves LR, Jacomé PRLA, Neto B, Pacífico J, Maciel MAV, et al. Biofilm production by clinical isolates of Pseudomonas aeruginosa and structural changes in LasR protein of isolates non biofilm-producing. Braz J Infect Dis. 2018;22(2):129–136.
    1. Bjarnsholt T, Jensen PØ, Jakobsen TH, Phipps R, Nielsen AK, Rybtke MT, et al. Quorum sensing and virulence of Pseudomonas aeruginosa during lung infection of cystic fibrosis patients. PLoS One. 2010;5(4):e10115.
    1. Senturk S, Ulusoy S, Bosgelmez-Tinaz G, Yagci A. Quorum sensing and virulence of Pseudomonas aeruginosa during urinary tract infections. J Infect Dev Ctries. 2012;6(6):501–507.
    1. Singh S, Singh SK, Chowdhury I, Singh R. Understanding the mechanism of bacterial biofilms resistance to antimicrobial agents. Open Microbiol. J. 2017;11:53–62.
    1. Abidi SH, Sherwani SK, Siddiqui TR, Bashir A, Kazmi SU. Drug resistance profile and biofilm forming potential of Pseudomonas aeruginosa isolated from contact lenses in Karachi-Pakistan. BMC Ophthalmol. 2013;13:57.

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