Streptococcus mutans was cultured in BHI broth for 24 h at 37°C with 5% CO2. After overnight growth, 1 mL of a standardized suspension containing 107 cells/mL was inoculated into 6 mL of BHI broth and incubated for 4 h at 37°C (5% CO2) (Barbosa et al., 2016 (link)). The culture was centrifuged at 5000 rpm for 10 min and the supernatant was filtered with a 0.22 μm diameter pore membrane using a vacuum filtration system (Stericup® and Steritop® Filter Unit, Millipore, MA, United States).
Free full text: Click here
