AimThe present study sought to compare the antimicrobial efficacy of 3% NaOCl, Biopure MTAD (Tulsa Dentsply, Tulsa, OK) and Brazilian ethanolic extract of propolis (EEP) against Enterococcus faecalis (E. faecalis).MethodologyThe study utilized 55 extracted human permanent teeth with single root canal. The samples were decoronated, instrumented and sterilized. The teeth were infected with E faecalis for 48 hours. The teeth were divided randomly into 3 groups according to the irrigants used and kept in contact with the respective irrigant for 5 minutes. All the samples were incubated in brain heart infusion (BHI) broth for 96 hours. Disinfection of the samples was determined based on presence or absence of turbidity in the BHI broth 96 hours later. Statistical analysis was done using Chi-square test.ResultsAll the samples treated with MTAD showed complete absence of turbidity, while all the 15 teeth treated with propolis showed presence of turbidity, 8 out of 15 teeth treated with NaOCl showed presence of turbidity. Statistical analysis of the data using chi-square test showed significant difference between the groups (P < 0.05).ConclusionThe study concluded that MTAD was more effective than 3% NaOCl and propolis against E. faecalis.