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Lactoferrin has amajor effects on enteropathogenes as it inhibits growth. so, This work adopted to study the antibacterial activity of lactoferrin on growth of E. coli non O157 in broth and pasteurized milk. Two strains of E. coli (O1 carry stx2 and hly gene and O26 carry hly gene) were used. Different concentrations of lactoferrin (zero, 0.5, 1.0, 5.0, 10 and 20 mg/ml) were used in Lauria broth. Based on our results, LF showed various inhibition activity on E. coli non O157 growth in Lauria broth. Significant decrease observed on growth of E. coli O1. Higher concentrations of lactoferrin caused a significant decrease in E. coli O26 growth. Regarding its effect in pasteurized milk, a significant decrease in the count of E. coli O1and E. coli O26 at the concentrations of 10 and 20 mg/ml lactoferrin was noticed. So, lactoferrin could become a promising method to decrease growth of E. coli non O157 in pasteurized milk consequently decrease E.coli non O157 associated illness in humans.