Food security is critical in the foodstuffs industry since most bacterial pathogens may cause foodborne illnesses and harm general health. In food, E. coli infection was recognized as the major cause of many cases of foodborne diseases. Moreover, the hazards have increased remarkably as most bacteria have acquired resistance to many antibiotics. Bacteriophages (phages) are a natural alternative agent that can be used for food preservation; they are viruses that can target bacteria specifically without causing harm to the human, plant, or animal cells. This research aimed to evaluate the efficacy of phage isolates in suppressing E. coli strains isolated from food samples. The phage was collected from sewage water and tested against E. coli isolate (EC/20) in vitro. Phage ZCECO 5 genome size was assessed to be ~339.5 kbp, and electron microscopy examination revealed that it related to the Myohoviridae family. In addition, it was found to be tolerant to a wide range of temperatures and pH. At different Multiplicity of Infections (MOIs), it showed a decrease in bacterial counts, with more bacterial elimination at stronger MOIs. Additionally, at MOI 10, the phage inhibited E. coli optical density from 1.0 OD600 to 0.7 OD600 after 230 min, and following a four-hour incubation period at 37°C, the bacterial titer decreased. The results indicated that ZCECO 5 could lyse E. coli and inhibit its activity. As a result, phage ZCECO 5 is suggested to be a bio-control agent in food for E. coli control.