Physiochemical and Genomic Characterization of Bacteriophage against Urinary tract Inficting Pseudomonas Aeruginosa to assess its Possibity for Phagr Therapy

Abstract

Introduction: Emerging antibiotic resistance against widespread Uropathogen has grown to be a significant therapeutic concern for UTI in recent years particularly in low- and middle-income nations by the reckless uses of antibiotics. In present context, resistance is seen in almost all antibiotics even in the last line Carbapenems and colistin. Thus, using phages in therapeutics to fight against antimicrobial resistance can be a solution to the global threat caused by these resistant bacteria. This study aims to isolate and characterize the phage both physiochemically and genetically to combat this global issue. Methodology: Host bacterial species was identified by biochemical test and 16SrRNA sequencing. Antibiotic susceptibility test was performed by disc diffusion method and confirmed by Vitek Compact System 2 analyzer. Bacteriophage isolation was done by Double Layer Agar Assay (DLAA). pH and temperature stability of phage was analyzed. Latent period, burst size and host range was determined. Whole genomic sequencing of phage was done. Bioinformatic analysis to assess for any kind of toxin gene or the virulence gene of the bacterial origin using different tools such as Ugene, RAST, PHASTER and Proksee. Results: Lytic bacteriophage against Pseudomonas aeruginosa was isolated. The phage was maximally stable at temperature 37˚c and pH 7. The latent period was 30 min and burst size was 96 virions per bacterium. Isolated phage 6661 showed intraspecific host range with Pseudomonas (P1) whereas no any activities with other Pseudomonas. Protein profiling of the phage through SDS- PAGE shows the four distinct band of protein in the gel. The size of DNA was found to be greater than 10kb. Phage length and GC content was determined by whole genome sequencing which was 43212 kb and 53.79% respectively. No any toxin or virulence gene was determined on bioinformatic analysis. The isolated phage significantly reduces the biofilm by 50.73%. Conclusion: Pseudomonas phage 6661 showed good stability to various physiochemical factors and on genomic characterization, presence of endolysin absence of toxin or virulence gene and integrase enzyme favors the therapeutic potential of the virus. Keywords: Antibiotic resistance, Bacteriophage, Burst size, Colistin, Phage Therapy