This study investigated the antibacterial efficacy against Streptococcus mutans and fluoride release of a conventional glass ionomer (GI) contained natural and chemical agents. Two hundred and ten GI specimens were divided into ten groups (n=21) according to the concentrations of the additives as; Propolis extract containing GI (Groups 1, 2, 3) with concentrations of 0.25%, 0.75% and 1.25% respectively, Miswak extract containing GI (Groups 4, 5, 6) and Chlorhexidine containing GI (Groups 7, 8, 9) with the same concentrations. The prepared specimens were subjected to antimicrobial activity by well diffusion, bacterial adherence, and fluoride release (from 2 to 72 hours) assessments. A higher statistically significant antibacterial activity was found in (Groups 2, 3) compared to (Groups 8, 9), while (Groups 1, 4, 5, 6, 7, 10) no antibacterial efficacy was reported. For (Groups 2, 3) had a higher statistically significant anti-adherence effect compared to the other tested groups. Enhanced ascending increase in fluoride release was observed for (Groups 3, 4) compared to (GI). The increased concentration of propolis extract had a synergistic effect on the antimicrobial activity of the tested GI. Additive concentrations of 0.25% Miswak and 1.25% propolis could enhance the fluoride-releasing ability of the tested GI.