Relationship between antibiotic resistance, biofilm formation, genes coding virulence factors and source of origin of Pseudomonas aeruginosa clinical strains
More details
Hide details
Chair and Department of Genetics and Pharmaceutical Microbiology, Poznan University of Medical Sciences, Poland
Microbiology Clinical Laboratory, University Hospital of the Lord’s Transfiguration, Poznań, Poland
Corresponding author
Magdalena Ratajczak   

Poznan University of Medical Science, Poland
Ann Agric Environ Med. 2021;28(2):306-313
Introduction and objective:
Pseudomonas aeruginosa is an opportunistic pathogen that causes difficult with treating infections, especially in the immunocompromised and patients with some underlying disease. The aim of the study is to assess the antibiotic resistance, biofilm formation, and the presence of genes encoding various virulence factors in clinical isolates of P. aeruginosa.

Material and methods:
Seventy-three clinical isolates of Pseudomonas aeruginosa were tested. Antimicrobial Susceptibility Testing (AST) and carbapenemases production was performed in accordance with the EUCAST guidelines. The ability to form biofilm was assessed by crystal violet assay. Genes encoding selected virulence factors were detected using standard polymerase chain reaction (PCR).

Among the 73 clinical isolates of P. aeruginosa, 41.1% were resistant to imipenem, 61.6% to meropenem, 30.1% to ciprofloxacin and 15.1% to tobramycin. Over 20% of isolates were producers of MBL. Antibiotic resistance profiling revealed that 23.3% of strains were sensitive to all antibiotics, 60.3% were LDR phenotype, and 16.4% were MDR phenotype. The majority of strains (73.6%) were strong-biofilm producers, 17.0% were moderate and 9.4% were weak biofilm producers. PCR analysis showed the presence of lasB, aprE and prpL genes in most of the tested strains (93.1%, 87.7% and 74.0%, respectively). Among strong biofilm producers, 22.2% were MDR, 63.0% of strains represented LDR phenotype, and 14.8% were sensitive to all antibiotics. Moderate and weak biofilm producers were LDR and sensitive phenotypes only (respectively, 58.3% and 42.9 – LDR, 41.7 and 51.7% – sensitive).

High frequency of MDR strains and their ability of biofilm formation and virulence factors may be a threat to effective therapy, and can increase morbidity and mortality of infected patients.

Pachori P, Gothalwal R, Gandhi P. Emergence of antibiotic resistance Pseudomonas aeruginosa in intensive care unit; a critical review. Genes Dis. 2019; 6: 109–19.
Tümmler B. Emerging therapies against infections with Pseudomonas aeruginosa. F1000Res. 2019.
Urbanowicz P, Gniadkowski M. „Ciężkozbrojny” Pseudomonas aeruginosa: mechanizmy lekooporności i ich tło genetyczne. Kosmos. 2017; 66: 11–29.
Yoon E-J, Kim D, Lee H, Lee HS, Shin JH, Park YS, Kim YA, Shin JH, Shin KS, Uh Y, Jeong SH. Mortality dynamics of Pseudomonas aeruginosa bloodstream infections and the influence of defective OprD on mortality: prospective observational study. J Antimicrobial Chemotherapy. 2019; 74: 2774–2783.
Bosaeed M, Ahmad A, Alali A, Mahmoud E, Alswidan L, Alsaedy A, Alalwan B, Alshamrani M, Alothman M. Experience With Ceftolozane-Tazobactam for the Treatment of Serious Pseudomonas aeruginosa Infections in Saudi Tertiary Care Center. Infect Dis: Res Treatment. 2020;
Hattemer A, Hauser A, Diaz M, Scheetz M, Shah N, Allen JP, Porhomayon J, El-Solh AA. Bacterial and Clinical Characteristics of Health Care- and Community-Acquired Bloodstream Infections Due to Pseudomonas aeruginosa. Antimicrobial Agents and Chemotherapy. 2013; 57: 3969–3975.
Kim YJ, Jun YH, Kim YR, Park KG, Park YJ, Kang JY, Kim SI. Risk factors for mortality in patients with Pseudomonas aeruginosa bacteremia; retrospective study of impact of combination antimicrobial therapy. BMC Infect Dis. 2014; 14–161. doi:10.1186/1471-2334-14-161.
Alhazmi A. Pseudomonas aeruginosa – pathogenesis and pathogenic mechanisms. Int J Biol. 2015; 7: 44–67.
Sabharwal N, Dhall S, Chhibber S, Harjai K. Molecular detection of virulence genes as markers in Pseudomonas aeruginosa isolated from urinary tract infections. Int Mol Epidemiol Genet. 2014; 5: 125–134.
Ghanbari A, Dehghany J, Schwebs T, Müsken M, Häussler S, Meyer-Hermann M. Inoculation density and nutrient level determine the formation of mushroom-shaped structures in Pseudomonas aeruginosa biofilms. Sci Rep. 2016; 9(6): 32097. doi: 10.1038/srep32097.
Mulcahy LR, Isabella VM, Lewis K. Pseudomonas aeruginosa biofilms in disease. Microb Ecol. 2014; 68: 1–12. doi:10.1007/s00248-013-0297-x.
Wang Y, Gao L, Rao X, et al. Characterization of lasR-deficient clinical isolates of Pseudomonas aeruginosa. Sci Rep. 2018; 8: 13344. doi: 10.1038/s41598-018-30813-y.
Hendiani S, Pornour M, Kashef N. Quorum-sensing-regulated virulence factors in Pseudomonas aeruginosa are affected by sub-lethal photodynamic inactivation. Pathodiagnosis and Photodynamic Terapy. 2019; 26: 8–12.
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: 501–507.
Vandeplassche E, Sass A, Lemarcq A, Dandekar AA, Coenye T, Crabbé A. In vitro evolution of Pseudomonas aeruginosa AA2 biofilms in the presence of cystic fibrosis lung microbiome members. Sci Rep. 2019; 9: 12859. doi: 10.1038/s41598-019-49371-y.
Yekani M, Memar MY, Alizadeh N, Safaei N, Ghotaslou R. Antibiotic Resistance Patterns of Biofilm-Forming Pseudomonas Aeruginosa Isolates from Mechanically Ventilated Patients. Int J Sci Study. 2017; 5: 84–88.
Kamali E, Jamali A, Ardebili A, Ezadi F, Mohebb A. Evaluation of antimicrobial resistance, biofilm forming potential, and the presence of biofilm-related genes among clinical isolates of Pseudomonas aeruginosa. BMC Res Notes. 2020; 13: 27.
Stefani S, Campana S, Cariani L, Carnovale V, Colombo C, Lleo MM, Iula VD, Minicucci L, Morelli P, Pizzamiglio G, Taccetti G. Relevance of multidrug-resistant Pseudomonas aeruginosa infections in cystic fibrosis. Int J Med Microbiol. 2017; 307: 353–362.
Potron A, Poirel L, Nordmann P. Emerging broad-spectrum resistance in Pseudomonas aeruginosa and Acinetobacter baumannii: Mechanisms and epidemiology. Int J Antimicrob Agents. 2015; 45: 568–85.
EUCAST, 2017. EUCAST guideline for the detection of resistance mechanisms and specific resistances of clinical and/or epidemiological importance. Version 9.0. files/Breakpoint tables/v9.0Breakpoint Tables.pdf.
Stepanovic S, Vukovic D, Hola V, Di Bonaventura G, Djukić S, Cirković I, Ruzicka F. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci, APMIS. 2007; 115: 891–899.
Cassini A, Högberg LD, Plachouras D, Quattrocchi A, Hoxha A, Simonsen GS, Colomb-Cotinat M, Kretzschmar ME, Devleesschauwer B, Cecchini M, Ouakrim DA, Oliveira TC, Struelens MJ, Suetens C, Monnet DL; Burden of AMR Collaborative Group. Attributable deaths and disability-adjusted life-years caused by infections with antibiotic-resistant bacteria in the EU and the European Economic Area in 2015: a population-level modelling analysis. Lancet Infect Dis. 2019; 19: 56–66.
Rosenthal VD, Rodrigues C, Álvarez-Moreno C, Madani N, Mitrev Z, Ye G, Salomao R, Ulger F, Guanche-Garcell H, Kanj SS, Cuéllar LE, Higuera F, Mapp T, Fernández-Hidalgo R, INICC members. Effectiveness of a multidimensional approach for prevention of ventilator-associated pneumonia in adult intensive care units from 14 developing countries of four continents: findings of the International Nosocomial Infection Control Consortium. Crit Care Med. 2012; 40: 3121–8.
Hong DJ, Bae K, Jang I-H, Jeong SH, Kang H-K, Lee K. Epidemiology and Characteristics of Metallo-ß-Lactamase-Producing Pseudomonas aeruginosa. Infect Chemother. 2015; 47: 81–97.
Marra AR, Pereira CA, Gales AC, Menezes LC, Cal RG, de Souza JM, Edmond MB, Faro C, Wey SB. Bloodstream infections with metallo-beta-lactamase-producing Pseudomonas aeruginosa: epidemiology, microbiology, and clinical outcomes. Antimicrob Agents Chemother. 2006; 50: 388–90.
Iwańska A, Nowak J, Skorupa W, Augustynowicz-Kopeć E. Analysis of the frequency of isolation and drug resistance of microorganisms isolated from the airways of adult CF patients treated in the Institute of Tuberculosis and Lung Disease during 2008–2011. Pneumonol Alergol Pol. 2013; 81: 105–13.
Cornaglia G, Giamarellou H, Rossolini GM. Metallo-ß-lactamases: a last frontier for ß-lactams? Lancet Infect Dis. 2011; 11: 381–93.
Silva LV, Galdino AC, Nunes AP, dos Santos KR, Moreira BM, Cacci LC, Sodré CL, Ziccardi M, Branquinha MH, Santos AL. Virulence attributes in Brazilian clinical isolates of Pseudomonas aeruginosa. Int J Med Microbiol. 2014, 304: 990–1000.
Jovcic B, Lepsanovic Z, Suljagic V, Rackov G, Begovic J, Topisirovic L, Kojic M. Emergence of NDM-1 Metallo-ß-Lactamase in Pseudomonas aeruginosa Clinical Isolates from Serbia. Antimicrobial Agents and Chemotherapy. 2011; 55: 3929–3931.
Pollini S, Mugnaioli C, Dolce D, Campana S, Neri AS, Taccetti G, Rossolini GM. Chronic infection sustained by a Pseudomonas aeruginosa High-Risk clone producing the VIM-1 metallo-ß-lactamase in a cystic fibrosis patient after lung transplantation. J Cyst Fibros. 2018; 17: 470–474.
Olejnízková K, Holá V. The comparison of selected virulence factors in Pseudomonas aeruginosa catheter isolates. Epidemiol Mikrobiol Imunol. 2012; 61: 21–8.
Lima JLDC, Alves LR, Jacomé PRLA, Bezerra Neto JP, Maciel MAV, Morais MMC. 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: 129–136.
da Silva Carvalho T, Rodrigues Perez LR. Impact of biofilm production on polymyxin B susceptibility among Pseudomonas aeruginosa clinical isolates. Infect Control Hosp Epidemiol. 2019; 40: 739–740.
Sharma G, Rao S, Bansal A, Dang S, Gupta S, Gabrani R. Pseudomonas aeruginosa biofilm: potential therapeutic targets. Biologicals. 2014; 42: 1–7.
Karatuna O, Yagci A. Analysis of quorum sensing-dependent virulence factor production and its relationship with antimicrobial susceptibility in Pseudomonas aeruginosa respiratory isolates. Clin Microbiol Infect. 2010; 16: 1770–5.
Kadhim D, Ali MR. Prevalence study of quorum sensing groups among clinical isolates of Pseudomonas aeruginosa. Int Curr Microbiol App Sci. 2014; 3: 204–215.
Pérez-Ibarreche M, Castellano P, Leclercq A, Vignolo G. Control of Listeria monocytogenes biofilms on industrial surfaces by the bacteriocin-producing Lactobacillus sakei CRL1862. FEMS Microbiol Lett. 2016; 363(12). doi: 10.1093/femsle/fnw118.
Mittal R, Sharma S, Chhibber S, Harjai K. Contribution of quorum-sensing systems to virulence of Pseudomonas aeruginosa in an experimental pyelonephritis model. J Microbiol Immunol Infect. 2006; 39: 302–9.
Vaněrková M, Mališová B, Kotásková I, Holá V, Růžička F, Freiberger T. Biofilm formation, antibiotic susceptibility and RAPD genotypes in Pseudomonas aeruginosa clinical strains isolated from single centre intensive care unit patients. Folia Microbiol. 2017; 62: 531–538.
Journals System - logo
Scroll to top