Molecular characterization of strong biofilm producing MDR, XDR and PDR Pseudomonas aeruginosa isolated from a tertiary care hospital in South India
Abstract Background : Pseudomonas aeruginosa is an opportunistic, gram negative bacterium that causes serious infections, especially among immunocompromised patients. An unusually high incidence of nosocomial P. aeruginosa infections was observed among patients hospitalized at Sri Sathya Sai Institute of Higher Medical Sciences, Puttaparthi, India, between 2014 November and 2015 February. Some of the patients being treated for a variety of cardiac and urological disorders in the Cardiothoracic and Vascular Surgery - Intensive Care Unit (CTVS-ICU) and Urology ward were found infected with P. aeruginosa . Active surveillance and environmental sampling revealed the presence of two additional MDR P. aeruginosa in the tap waters of CTVS-ICU. Based on the Antibiotic sensitivity pattern, fourteen P. aeruginosa MDR (n=7), XDR (n=6) and PDR (n=1) isolates (inclusive of the two tap water isolates) were shortlisted for additional investigations. These isolates were characterized for antibiotic sensitivity, biofilm production, minimum biofilm inhibitory concentration (MBIC), presence of antibiotic resistance genes, efflux pumps, and integrons. Results : Mutations in gyr A, gyr B, par C, mex R, nfx B, mex B and mex F genes were correlated to enhanced antibiotic resistance. Notably, the isolates were also found to harbor integrons and blaNDM-1. Pulsed Field Gel Electrophoresis (PFGE) and Random Amplified Polymorphic DNA (RAPD) based phylogenetic analyses grouped the clinical isolates into three distinct ward specific clusters, while the tap water isolates were grouped into a separate cluster. Detection of nosocomial P. aeruginosa in the CTVS-ICU and urology ward triggered the activation of enhanced surveillance and infection control measures to contain and eliminate P. aeruginosa infections. Conclusions : Multiple clones of P. aeruginosa are prevalent in the study center. Hence, continuous screening and identification of potential reservoirs is absolutely essential to control the spread of drug resistant P. aeruginosa infections.