Isolation and characterization of a novel lytic phage Artin targeting typhoidal and nontyphoidal Salmonella and evaluation of its antibacterial efficacy in food

Phages are increasingly viewed as powerful natural tools for controlling Salmonella and other foodborne pathogens, offering an alternative to conventional antimicrobials. However, the extensive serotype diversity of Salmonella and the rise of phage-resistant strains highlight the need for isolating phages with broader host ranges and strictly lytic activity. Advancing such phage resources is essential for developing reliable biocontrol strategies. In this study, a novel lytic phage with significant potential to combat various Salmonella strains was isolated from dog feces. Morphological and biological characteristics of phage Artin were assessed using transmission electron microscopy, stability assays under different temperature and pH conditions, and adsorption assays. Structural proteins were analyzed by SDS-PAGE. The host range was determined against four Salmonella serovars and six non-Salmonella species. Whole-genome sequencing followed by bioinformatics analysis was performed to elucidate the genomic features of Artin. The ability of phage Artin to control Salmonella Enteritidis in chicken breast meat and milk was further assessed at 4 °C. Statistical analysis was carried out using GraphPad Prism 8. 0. Genomic analysis classified SalmonellaₚhageArtin within the Caudoviricetes class, Markadamsvirinae subfamily, and Epseptimavirus genus, with a dsDNA genome size of 118, 477 bp and a GC content of 39. 78%, featuring 192 coding sequences (CDS) devoid of antimicrobial resistance, virulence factors, and toxins. Phage Artin showed strong stability over a wide range of temperatures (4–70 °C) and pH levels (2–12). Approximately 53% of phage particles adsorb to host cells within 5 min. In addition to S. Enteritidis, phage Artin was also able to infect S. Typhimurium, S. Typhi, S. Choleraesuis, and Escherichia coli. At 4 °C, Artin reduced recoverable Salmonella in chicken breast by 1. 3 log10 CFU/mL and 2. 7 log10 CFU/mL at MOIs of 10 and 1000, respectively, relative to the phage-free control. Also, the recoverable Salmonella in milk decreased by 1. 6 log10 CFU/mL at MOI of 10 and 3. 1 log10 CFU/mL at MOI of 1000 compared to the control. Phage Artin is a strictly lytic phage with broad host range and high stability, effectively reducing S. Enteritidis in chicken and milk, highlighting its potential as a safe and efficient biocontrol agent.