OBJECTIVE: The objective of this study was to evaluate the antibacterial efficacy of two doses of vetiver and chamomile essential oils compared with chlorhexidine and calcium hydroxide against Enterococcus faecalis. MATERIALS AND METHODS: The growth inhibition and minimum inhibitory concentration of all tested materials were determined ex vivo following agar diffusion and broth dilution assay procedure. Human maxillary anterior teeth were prepared with protaper rotary files, followed by incubation with standard broth of E. faecalis. A total of 140 teeth were included in the study. These teeth are randomized and equally divided into seven groups and were treated with low (1.25 µl) and high doses (2.5 µl) of essential oils of vetiver and chamomile and calcium hydroxide (0.1/1.0 mL), 2% chlorhexidine (2.5 µl). Microbial sampling of six teeth from each group was done with paper points and Gates-Glidden burs at 1, 7, and 14 days and colony-forming unit (CFU)/mL was determined. RESULTS: There was a significant reduction in mean ± standard deviation of CFU (log10) in vetiver oil high dose (3.32 ± 0.036) and chlorhexidine (3.34 ± 0.030), followed by calcium hydroxide (3.46 ± 0.015) and chamomile oil high dose (3.48 ± 0.20) on day 1. On 7th day, a significant reduction was seen in chlorhexidine (2.74 ± 0.212), chamomile oil (2.81 ± 0.035, low dose and 2.97 ± 0.119, high dose), followed by calcium hydroxide (3.25 ± 0.028). However, on 14th day, it was 2.32 ± 0.088 for chlorhexidine, 2.91 ± 0.029 for chamomile oil high dose, 3.15 ± 0.010 for vetiver oil high dose, and 3.09 ± 0.068 for calcium hydroxide. CONCLUSION: The study showed a good effectiveness of chamomile oils in root canal infection of E. faecalis at different time intervals compared to chlorhexidine and calcium hydroxide vetiver oil did not sustain their activity for a longer duration.