Isolation and characterization of a bacteriophage and its utilization against multi-drug resistantPseudomonas aeruginosa-2995

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Aims:To identify, isolate, and characterize a lytic bacteriophage against the multiple-drug resistant clinical strain of Pseudomonas aeruginosa-2995 and to determine the phage efficacy against the bacterial planktonic cells and the biofilm.Main methods:Wastewater was used to isolate a bacteriophage. The phage was characterized with Transmission electron microscopy (TEM). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) was used to identify the expressed proteins. Bacteria were cultured in both suspension and biofilm to check and compare their susceptibility to phage lytic action. The activity of the phage (determined as AZ1) was determined against P. aeruginosa-2995 in both planktonic cells and the biofilm.Key findings:A bacteriophage, designated as AZ1, was isolated from waste water showing a narrow host range. AZ1 was characterized by TEM and could be identified as an isolate in the family Siphoviridae [order Caudovirals]. Seventeen structural proteins ranging from about 12 to 110 kDa were found through SDS-PAGE analysis. Its genome was confirmed as dsDNA with a length of approx. 50 kb. The log-phase growth of P. aeruginosa-2995 was significantly reduced after treatment with AZ1 (4.50 × 108 to 2.1 × 103 CFU/ml) as compared to control. Furthermore, phage AZ1 significantly reduced 48 h old biofilm biomass about 3-fold as compared to control.Significance:Pseudomonas aeruginosa is a ubiquitous free-living opportunistic human pathogen characterized by high antibiotic tolerance and tendency for biofilm formation. The phage, identified in this study, AZ1, showed promising activity in the destruction of both planktonic cells and biofilm of P. aeruginosa-2995. However, complete eradication may require a combination of phages.

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