Synergistic Effects of Gentamicin, Cefepime, and Ciprofloxacin on Biofilm of Pseudomonas aeruginosa

Muhammad Usman,1 Arooj Marcus,1 Aimen Fatima,1 Bushra Aslam,1 Maryam Zaid,1 Muska Khattak,1 Sidra Bashir,1 Safia Masood,1 Zara Rafaque,2 Javid Iqbal Dasti1 1Lab of Microbial Genomics and Epidemiology, Department of Microbiology, Quaid-I-Azam University, Islamabad, 45320, Pakistan; 2Department of Microbiology Hazara University Mansehra, Mansehra 21120, PakistanCorrespondence: Javid Iqbal Dasti, Lab of Microbial Genomics and Epidemiology, Department of Microbiology, Quaid-I-Azam University, Islamabad, 45320, Pakistan, Tel +92-5190644175, Email iqbal.78@hushmail.comBackground: Pseudomonas aeruginosa is an opportunistic pathogen involved in number of hospital-acquired infections such as catheter-associated urinary tract infections, bacteremia, septicemia, skin infections, and ventilator-associated pneumoniae. Biofilm formation is an important trait implicated in chronic infections, such as cystic fibrosis and chronic pulmonary obstruction. We evaluated effects of gentamicin, cefepime, and ciprofloxacin on biofilm of P. aeruginosa.Materials and Methods: A total of 266 isolates were collected from the Armed Forces Institute of Pathology (AFIP). Antibiotic susceptibility was assessed by double disk synergy testing. ESBL and carbapenemase detection was performed by phenotypic testing. Molecular screening of the genes was done by PCR. Micro-dilution broth method was used to determine minimum inhibitory concentrations of antibiotics. Biofilm formation was done by micro-titer plate assay.Results: Overall, 20% of the P. aeruginosa isolates were extensively drug-resistant (XDR-PA), and 25% were multi-drug-resistant (MDR-PA). Likewise, 43% of the isolates were ESBL producers, and carbapenemase production was detected in 40% of the isolates. Molecular analysis confirmed occurrence of different resistant factors in ESBL-positive isolates; 67% carried blaTEM, 62% blaCTXM-15, 41% blaSHV, 34% blaCTXM-14, and 33% blaOXA-1. In addition, 68% of the carbapenem-resistant isolates were positive for blaNDM-1, 25% for blaOXA-48, and 22% for blaKPC-2. Biofilm formation was assessed for 234 isolates, out of which 28% were strong biofilm formers. Moderate and weak biofilm formers constituted 46% and 23%, respectively. Overall, ciprofloxacin, levofloxacin, and cefepime showed inhibitory effects on P. aeruginosa biofilms. Antibiotics in combination showed strong synergistic effects (ciprofloxacin and cefepime), while gentamicin and cefepime resulted in complete eradication of P. aeruginosa biofilm.Conclusion: We confirm strong synergistic effects of gentamicin and cefepime that completely eradicated P. aeruginosa biofilm. We further confirm inhibitory effects of ciprofloxacin, levofloxacin, and cefepime on P. aeruginosa biofilms. Hence, combination therapy can be more effective against biofilm-associated infections.Keywords: synergistic effects, gentamicin, cefepime, biofilm, Pseudomonas aeruginosa