Background: Virulence and resistance ability of Enterococcus faecalis caused a failure in endodontic treatment. Virulence of bacterias are supported by energy metabolic activities. One of energy metabolic source is arginine. Propolis as an herbal medicine has an antibacterial action with complex mechanisms. Objective: This study aims to determine the effect of ethanolic extract of propolis (EEP) from Apis Trigona on arginine metabolic activity of Enterococcus faecalis. Method: The design of this study is an in vitro experimental laboratory that begins with a preliminary. The method used was the arginine metabolism test in Arginine Dehydrolase broth media which was preceded by pretreatment in clinical bacterial samples and ATCC 29212, followed by color media quantification using imageJ and media pH measurements. EEP tested on bacterial cultures consisted of various concentrations: 0.00125%, 0.4%, 10% with positive control of chlorhexidine digluconate 2%. Results: The color observation results in all EEP concentrations had a color changed in the media, while positive controls had no color changed in media. Color changed indicate the arginine metabolic activity. Data analyze of imageJ quantification by OneWayANOVA test and Krusskal-Wallis test, both showed that all EEP concentrations had a significant difference with positive controls (p<0.05) which indicates that all EEP concentrations cannot inhibit arginine metabolic activity. Conclusion: Ethanolic extract of propolis (Apis Trigona) has no effect on arginine metabolic activity of Enterococcus facealis.