In our study, we examined the effect of kutajarishta and its constituents on five strains of E.coli, one of the causative organisms of colitis. Colitis is an inflammation of colon caused due to micro-organisms like E.coli, Shigella Campylobacter, Salmonella and Yersinia sp. Our study deals with the self-generated herbal fermentation of traditional ayurvedic formulations for colitis the Arishtas and Asavas. They are alcoholic medications prepared by allowing herbal juices or their decoctions to undergo fermentation along with dhataki flowers (Woodfordia fruticosa) and sugars. Dhataki flowers are fermentation initiators. Kutajarishtas also an ayurvedic formulation of Holarrhena sp (kudasaal), Gmelina arborea (Shivan), Woodfordia fruticosa (dhataki flowers), water, raisins and jaggery and can be used to treat colitis. To test the antimicrobial activity of kutajarishta, five strains of E.coli were isolated and confirmed by Indole, Methyl red, Voges-Proskauer and Citrate utilization (IMViC) tests. Alcoholic and aqueous extract of kudasaal and shivan were prepared and the effect of alcoholic and aqueous extract of kutajarishta, kudasaal and shivan were seen against five strains of E.coli by using agar well diffusion method. Further, compound purification from aqueous and alcoholic extract was done by TLC. Kutajarishta is most effective against all the five strains of E. coli as compared to kudasaal and shivan plant extracts. From the study, it can be concluded that the Kudasaal plant extract is more effective than shivan plant extract. Aqueous extracts of kudasaal and shivan are more effective than alcoholic kudasaal and shivan extracts. From the initial screening of all the three extracts we concluded that kutajarishta has maximum antibacterial activity due to the interaction of toxins with the phyto-constituents of the plant.