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Spot assay was conducted to evaluate the antibacterial effect at different pH values [22]. S. mutans (106 CFU/mL) in buffered BHI broth with or without LH12 was incubated at 37°C anaerobically for 5 min, 15 min and 30 min. Aliquots (15 μL) were cultured on agar plates at 37°C anaerobically for 24 h. The experiment was performed in triplicate and repeated three times.
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