The complexity of the canal system makes it impossible to fully shape and clean with the endodontic instruments alone should be combined with disinfection protocol. As a result, Root canal disinfection is an exceedingly tough process that results in vast exposed regions and residual biofilms of bacterial species in the root canal. AIM: Using microbial collection and culture methods, the study investigated the effect of different irrigation systems on the reduction of Enterococcus faecalis (E. faecalis) Materials and Methods: E. faecalis was inoculated into sterilized human teeth. To irrigate the samples 30-gauge double-vented needle, an Endoactivator, and a conventional needle were used. After chemomechanical treatment, dentin fragments from the apical third of the canal were collected by Gates Glidden 1. Estimated colony forming unit (cfus) and incubation of E. faecalis per mg dentin after addition of 1 mL of sterile Brain infusion (BHI) suspension for the sample. RESULTS: The decrease in Enterococcus faecalis was seen in the experimental group compared to other groups (p < 0.05). Group I and Group III displayed a significant difference (P<0.05). CFU/ml counts between all three groups showed statistically significant differences CONCLUSION: The Endoactivator and double-side vented needle were better in bacterial reduction compared to conventional needle irrigation