Isolation and Genomic Characterization of a Lytic Bacteriophage Against Multidrug-Resistant E. coli

Antimicrobial resistance (AMR) has become a major concern in the treatment of bacterial infections, and bacteriophage therapy has emerged as a promising alternative to antibiotics. Bacteriophages are highly specific to their bacterial hosts; hence, isolating phages indigenous to a specific region offers a significant advantage against various pathogen strains. We have isolated a cocktail of bacteriophages against pathogenic E. coli from sewage water at a primary healthcare centre. Characterisation of the isolated phages demonstrated their stability across a broad pH and temperature range, strong lytic activity, and effective biofilm degradation, with no cross-reactivity with Staphylococcus aureus (S. aureus). Genomic analysis and phylogenetic studies indicated that the largest phage (by genome size) in the cocktail belongs to the genus Vequintavirus (myoviruses, rV5-like phages), and its genome sequence has been deposited in NCBI (Accession ID: PX741096). The phage genome was linear, with headful (PAC) packaging, encoded lysis proteins, and lacked antibiotic-resistant or major lysogeny-associated genes, collectively suggesting a lytic lifestyle. These findings emphasize the therapeutic potential of rV5-like phages and underscore the critical need to establish phage banks in India to improve disease management.