Escherichia coli is a Gram negative bacillus belonging to Enterobacteriaceae family, causing a plethora of human infections like Urinary tract infection (UTI), meningitis, diarrhoea and others. It is often resistant to multiple antibiotics. High cost of antibiotics, cost of prolonged hospital stay and refractoriness to drugs necessitates the invention of new, natural and low-cost compounds that can be effective against it. We here report the efficacy of extract of Neem leaves in inhibiting growth, bioflm formation and phospholipase (lecithinase), an important virulence determinant of E. coli. Furthermore, the extract was completely non-toxic to humnan RBCs and WBCs.