Background and objectives: This study evaluated and compared the antimicrobial efficacy and minimal inhibitory concentration (MIC) of chlorhexidine (CHX) 0.12% and extracts of aloe vera and mushroom against Streptococcus mutans and Lactobacillus. Materials and methods: The agar disk diffusion and the broth microdilution method were used to check the antimicrobial activity of 0.12% CHX and aqueous extracts of two medicinal plants. The test samples were divided as follow: S. mutans, group I: CHX, group II: aloe vera, and group III: mushroom. Lactobacillus, group I: CHX, group II: aloe vera, and group III: mushroom. The zone of inhibition and MIC values were tabulated and the data were statistically analyzed using ANOVA and Bonferroni post hoc tests. Results: Chlorhexidine shows maximum antibacterial action against S. mutans and Lactobacillus followed by the mushroom extract and then aloe vera extracts with the zones of inhibition of (20.3 mm/24.13), (17.03/16.05), and (14.09/14.26), respectively. Both the extracts had MIC value of 80 μg/mL and CHX was 40 μg/mL. Conclusion and inference: Within the limitations of this in vitro study, it can be concluded that all the herbal extracts tested in this study demonstrated antibacterial activity against mutans streptococci (MS) and Lactobacillus. Chlorhexidine shows maximum antibacterial action against S. mutans and Lactobacillus followed by the mushroom extract and then aloe vera extracts with the zones of inhibition of (20.3 mm/24.13), (17.03/16.05), and (14.09/14.26), respectively. Herbal products have potent antimicrobial activity that can be looked at as an alternative to CHX. However, further in vitro and long-term in vivo studies are recommended to confirm and correlate the findings of this study to a clinical situation.