Aim Assessment of the antimicrobial efficacy of zinc oxide eugenol root canal sealer and the value of incorporating amoxicillin in the sealer against Enterococcus Feacalis in extracted teeth. Methodology Seventy eight extracted human single rooted teeth were infected for with E. faecalis. Some specimens were shaped by Stainless steel (St. St.) instruments and others shaped by ProTaper instruments. Following mechanical root canal preparation, each group was divided according to sealer composition into subgroups as follows; Sub group 1 obturated using zinc oxide-eugenol root canal sealer and gutta percha. Sub group 2, amoxicillin was incorporated in the sealer composition. The obturated extracted teeth were incubated in Brain-Heart Infusion (BHI) broth up to 6 months. During incubation, specimens were submitted for bacterial monitoring at various obturation periods. Specimens were sampled before mechanical preparation, after mechanical preparation and after obturation. Bacterial monitoring was performed via serial dilution then culturing onto BHI agar plates. Colony forming units (cfu) were counted and percent reduction was calculated. Results sealers reduced significantly E. faecalis counts. Incorporating amoxicillin in sealing the Stainless steel shaped specimens, relatively enhanced the antibacterial sealing outcomes. Conclusions Root canal obturation using all tested sealers were profound antibacterial measures to eradicate E. faecalis. However, it cannot totally eliminate E. faecalis from infected root canals.