(Background): Salmonella spp. are significant foodborne pathogens that pose a serious risk to public health. These bacteria are commonly associated with contaminated food, particularly raw or undercooked poultry, eggs, and dairy products. Salmonella infections can lead to gastroenteritis, fever, and in severe cases, hospitalization and even death. Bacteriophages, which are viruses that specifically target and kill bacteria, have emerged as a promising strategy to control foodborne contaminants. They offer a natural and environmentally friendly alternative to traditional antimicrobial agents. By employing bacteriophages as biocontrol agents, the risk of Salmonella transmission through food can be effectively reduced, enhancing food safety and protecting public health.

(Methods): Salmonella host strain ZC_S-30 was isolated and characterized from food samples, and its antibiotic profile and virulence genes were detected. A lytic phage vB_Sal_ZC-10 was isolated from environmental water sewage samples on Salmonella host strain ZC_S-30. The isolated phage underwent purification and amplification, and its molecular DNA was assessed using Pulsed Field Gel Electrophoresis (PFGE). Additionally, Transmission Electron Microscopy (TEM) was employed to visualize its structure and morphology, while antibacterial activity against various Salmonella spp. hosts was evaluated. Stability tests were conducted, and the entire genome of the phage was sequenced. The efficiency of phage suspension was evaluated in vitro and in milk inoculated with Salmonella spp. host strain.

(Results): Phage vB_Sal_ZC10 was found to be belonging to siphoviruses as indicated by the TEM. The PFGE and the phage sequencing estimated the phage genome size of 48 kbp. In addition, the results indicated that phage vB_Sal_ZC10 withstands high stability at different temperatures (-80 up to 80 °C) and pH ranges (pH 3 - 9). For the antibacterial activity, phage vB_Sal_ZC10 exhibited clear zones on its ZC_S-30 bacterial host along with other host strains. In addition, the in vitro results of the time killing tests showed a significant bacterial eradication over different periods of time at varying MOIs (0.001, 0.01, 0.1, 1, 10, 100). Additionally, phage-treated milk inoculated with Salmonella host strain showed a biocontrol efficiency by reducing the bacterial titer by a minimum of 2 Log₁₀ CFU/ml at MOI of 100 and below the detection limit with MOI of 100,000 over the time of experiment.