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Abstract The purpose of this study was to undergo a bio-activity guided fractionation of four medicinal plants; namelyNeem(AzadirachtaindicaA.Juss), Ginger (Zingiberofficinale Roscoe), Miswak(Salvadorapersica L.) and Lemon(Citrus limonumRisso). The four most active fractions where then used as root canal irrigants, against single species of Enterococcus faecalis, using NaOCl as a positive control. The antimicrobial sensitivity tests in this study included; the Agar well- diffusion method (through the observation of the zone of microbial growth inhibition diameters), the Minimum Inhibitory Concentration (MIC) test which was determined by micro broth dilution technique to determine the most effective concentration to be used, and the Direct Contact Test, through testing the in-vitro intracanal microbial reduction, using the most potent medicinal plant fractions. The plant extract fractions that exhibited antimicrobial activity against Enterococcus faecalis as revealed by the agar well diffusion test were; Ethyl acetate fraction of Neem, ethyl acetate, butanol fractions and total ethanolic extract of Ginger, ethyl acetate fraction of Miswak and fresh Lemon solution. The direct contact test results revealed that (5.25% NaOCl) was associated with less CFU/ml count when compared to the tested plant extracts. Fresh Lemon solution was associated with less CFU/ml count when compared with all of the tested plant extracts followed by butanol fraction of Ginger, ethyl acetate fraction of Miswak and Neem |