Isolation and Characterization of Bacteriophage Against Wastewater Isolates of Methicillin-resistant Staphylococcus aureus

Background: Methicillin-resistant Staphylococcus aureus (MRSA) not only is a notorious pathogen in clinical settings but also is an environmental issue that its presence in environmental wastewater is highlighted by several reports. Due to the negative impacts of antibiotics, alternatives like bacteriophages, as biocontrol, are considered safe. However, not all bacteriophages are safe. Thus, the characterization of bacteriophages is necessary. Objectives: This study aimed to, firstly isolate MRSA from wastewater and, secondly to perform bacteriophage isolation from the water samples to investigate its physical and genomic characteristics. Methods: Water samples were collected from seven locations across Nagpur city, India, bacteria were isolated on the S. aureus specific agar. For detecting MRSA, we followed the disc diffusion method. Isolation of bacteriophage against MRSA was performed by a modified enrichment method. We investigated its physical characteristics by the one-step growth rate, adsorption rate, host range, survivability, electron microscopy, and genomic sequencing for bioinformatics analysis. Results: Four MRSA were isolated from wastewater samples. We got a bacteriophage against an MRSA from the river Ganga. The bacteriophage belongs to the Podoviridae family, subfamily Autographivirinae. It was stable till 40°C and could survive at a highly alkaline pH. It is specific to its host. The bacteriophage DNA encodes 52 ORF, and all predicted genes are on the same strand; it also encodes a phage RNA polymerase. Conclusions: It is the first report of an S. aureus bacteriophage that belongs to the sub-family Autographivirinae. Our study and literature survey conclude that S. aureus bacteriophages of the Podoviridae family are safe for various downstream applications.