Background: Dental caries is the most widespread disease affecting human race where Streptococcus mutans and Lactobacillus acidophilus are prime microorganisms involved. Chlorhexidine is used as antiplaque agent, but it has some side effects such as unpleasant taste and staining on long-term use, leading to search of a suitable alternative. Chicory, an ayurvedic plant, has antiplaque, anti-inflammatory, and healing properties along with aromatic cooling property. Aim: This study aims to evaluate the antimicrobial efficacy of chicory extract mouthwash and chlorhexidine mouthwash (0.12%) against S. mutans and L. acidophilus. Methods: Strains of S. mutans and Lactobacillus were grown and subjected to test solutions (chicory and chlorhexidine) and zone of inhibition measured by agar well diffusion method. Results obtained were subjected to statistical analysis. Results: Zone of inhibition of chicory extract and chlorhexidine against S. mutans and Lactobacillus shows highly significant P < 0.001 with +value 19.982 and 21.731, respectively. Conclusion: Chicory extract showed statistically significant inhibition against S. mutans as compared to chlorhexidine rinse mouthwash. While chlorhexidine mouthwash has shown statistically significant inhibition against S. mutans as compared to chicory extract. Clinical Significance: Chicory has a promising antibacterial activity. Thus, it can be incorporated as one of the ingredients in mouthwash.