Isolation and Characterization of Bacteriophage from Sewage and its Therapeutic Potential for Multidrug Resistant Escherichia Coli on Mice

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The increasing incidence of antibiotic resistance, the emergence of virulent bacterial pathogens, and lack of new effective antibiotics, has increased interest in the use of lytic bacteriophage therapy. The aim of this study was to characterize colliphage isolated from sewage to determine their in vitro and in vivo application. This experimental study was conducted at National Veterinary Institute, Bishoftu, Ethiopia from October 2019 to May 2020. Five different locations in Bishoftu were selected for the collection of fecal and sewage samples for the isolation of E.coli and detection of probable phages. After isolation of seven different E.coli isolates antibiotic susceptibility tests were conducted and majority of the isolates were resistant to more than five out of eight drugs. Bacteriophage were isolated from sewage samples and the isolated phage were purified by diluting in the buffer and filtered through 0.22 ��m filter. Purified lysate was further processed for analyzing its host range by using spot method. From these all seven isolates of E.coli only one isolate (E.coli 7) was susceptible to isolated. The Plaque forming unit of was maximum at pH 7, while the reduction was observed at both pH less than 5 and greater than 9. No plaques were observed at pH 3 and 12. The Plaque forming unit (PFU) was highest after treatment at 37°C while at 4oC there is no plaque formation, when the temperature increase from 4oC to 37oC the PFU increase and then decrease from 37oC to 42oC however no lytic activity was observed after treatment above 42°C. The effect of chloroform on phage activity was cheeked then the phage produce plaque before and after the addition of chloroform. 71.4% mice have survived receiving phage therapy. However 85.7% of the mice were died in the group that only E. coli got. Isolation, characterization and therapeutic application of potential phages lytic against E.coli bacteria commonly involved in infections is a major finding of this study. Notably, we found phage lytic against multidrug-resistant E.coli. This promising effect against MDR pathogens has raised the probable utility of these phages for biological control of bacterial infection. Further characterization of specific phages is needed to explore the potential use of these phages for their clinical application



Bacteriophage, in vitro, in vivo, MDR E. coli, mice, therapy