Characterization of Newly Isolated Rosenblumvirus Phage Infecting Staphylococcus aureus from Different Sources

Staphylococcus aureus is a globally significant pathogen associated with severe infections, foodborne illnesses, and animal diseases. Its control has become increasingly challenging due to the spread of antibiotic-resistant strains, highlighting the urgent need for effective alternatives. In this context, bacteriophages have emerged as promising biocontrol agents. This study aimed to characterize the newly isolated Staphylococcus phage CapO46 and evaluate its efficacy in reducing S. aureus in milk. Identified as a new species within the Rosenblumvirus genus, CapO46 exhibited a podovirus-like structure and a small linear dsDNA genome (17,107 bp), with no lysogeny-related, antimicrobial resistance, or virulence genes. Host range assays demonstrated its ability to infect all 31 S. aureus isolates from two different countries and in diverse environmental contexts, achieving high efficiency of plating (EOP > 0.5) in 64.5% of cases. Kinetic analyses revealed rapid adsorption and a short latent period, with a burst size of approximately 30 PFU/cell. In UHT whole-fat milk, CapO46 achieved a maximum reduction of 7.2 log10 CFU/mL in bacterial load after 12 h, maintaining significant suppression (1.6 log10 CFU/mL) after 48 h. Due to its genetic safety, high infectivity across multiple isolates, and antimicrobial activity in milk, CapO46 can be considered a promising candidate for S. aureus biocontrol applications.