The Moringa oleifera can be used as a medium for the prevention and treatment of infectious diseases such as diarrhea. Diarrhea can be caused by Escherichia coli, which house flies can transmit. Moringa oleifera leaves have antimicrobial substances such as flavonoids, triterpenoids, steroids, saponins and tannins. This aim of the study was to determine the minimum inhibitory concentration of M. oleifera infusion on the growth of E. coli. This research is a laboratory experiment with Well Diffusion Agar method and dilution technique. The materials used in the study were M. oleifera leaf infusion with concentrations of 50%, 40%, 30%, 20%, and 10%, while 0% was a negative control (Aquadest). E. coli isolates were obtained from M. domestica. The results showed that the inhibition zone formed at concentrations of 50%, 40%, 30%, 20%, 10%, and 0% was 15.55 mm, 13.80 mm, 13.10 mm, 10.10 mm, and 0 mm, respectively. The minimum inhibitory concentration of leaf infusion against E. coli isolates was 25%, and the minimum bactericidal concentration could not be determined because all concentrations tested showed bacterial growth. Instead, the lowest inhibitory concentration was 25%. In concentrations of 50%, 40%, and 30%, M. oleifera leaf infusion has potent antibacterial action; at concentrations of 20%, it has moderate antimicrobial activity; and at concentrations of 10%, it is unable to inhibit the growth of E. coli. Further research is needed to determine the value of the minimum bactericidal concentration at concentrations above 25%, and it is necessary to test it on other digestive tract pathogenic bacteria.