Cultivation and Maintenance of Oral Bacteria

The Enterococcus faecalis (CCARM 5511), Enterococcus faecium (KACC 11954), Porphyromonas gingivalis (KCTC 5352), Streptococcus mutans (KACC16833), Streptococcus sobrinus (KCTC5809), Staphylococcus aureus (CCARM3506), Staphylococcus epidermidis (KACC 13234), Fusobacterium nucleatum (KCTC 2640) and Actinomyces viscosus (KCTC 9146) strains used in this study are listed in Table 1. E. faecalis, E. faecium, S. epidermidis, S. aureus and A. viscosus were maintained in tryptic soy broth (TSB). S. mutans and S. sobrinus were maintained in brain heart infusion broth (BHI). P. gingivalis was maintained in tryptic soy broth supplemented with 10% defibrinated horse blood. F. nucleatum was maintained in reinforced clostridial agar and incubated at 37 °C. A. viscosus, S. sobrinus, F. nucleatum and P. gingivalis bacteria were cultured in brain heart infusion agar plate (BHI), reinforced clostridial agar plate, tryptic soy agar plated (TSB) and blood agar plates (Blood Agar Oxoid No₂; Oxoid, Basingstoke, UK), supplemented with 5% (v/v) sterile horse blood (Oxoid) at 37 °C for 24–72 h in anaerobic conditions (10% H₂, 10% CO₂, and balanced N₂) [21 (link),22 (link),23 (link)].