WITHDRAWN: SYNERGISM OF BACTERIOPHAGE AND ANTIBIOTICS IN THE REDUCTION OFPseudomonas aeruginosaBIOFILM

Abstract

IntroductionPseudomonas aeruginosa is resistant to most of the antibiotics. This makes treatment of Pseudomonas aeruginosa difficult. The problem is further compounded by its ability to form biofilm. The aim of this study was to isolate lytic phage against antibiotic resistant Pseudomonas aeruginosa and to use it in the reduction of Pseudomonas aeruginosa biofilm.MethodologyBacteriophage isolation was done by Double Layer Agar Assay method. Phage cocktail was used to disrupt biofilm. The synergistic effect of phage and antibiotic in reducing biofilm was analyzed. Effect of various external factors in phage stability was examined. Whole genome sequencing of phage DNA was done.ResultLytic bacteriophage against Pseudomonas aeruginosa was isolated. Three distinct bands of phage proteins of size 35, 40 and 100KDa were observed after performing SDS PAGE. Phage DNA size was determined to be larger than 10 Kb from agarose gel electrophoresis. Whole genome sequencing of phage revealed its size to be 43,428 nucleotides (43 Kb) in length and the GC content of 62.16%. There was no presence of virulence gene in the phage genome and it contained holin and endolysin gene.Calcium ion increased the phage adsorption. Phage showed stability against SDS and osmotic shock whereas it was susceptible to ethanol, acetone and CTAB. Phage TU_pse1B reduced the biofilm by 60.99% whereas Phage TU_pse1Bi reduced the biofilm by 60.37%. Synergism of phage and antibiotic was observed in reducing biofilm. Phage plus antibiotic reduced biofilm by further 26.67% than phage alone.ConclusionPhage TU_pse1B showed good stability to various physiochemical factors as well as it was efficient in reducing Pseudomonas aeruginosa biofilm. So, this phage can be a good candidate for controlling antibiotic resistant Pseudomonas aeruginosa.