Objective: Microorganisms reside in polymer-coated communities known as biofilms, which represent a major virulence factor by conferring resistance to treatment in associated infections. Biofilms are therefore a frequent subject of health-related research. However, standard laboratory strains have been cultured under inconsistent conditions, with limited data available on optimal biofilm production parameters. This study aimed to determine the optimal in vitro biofilm formation conditions for Streptococcus mutans ATCC 25175 and Staphylococcus epidermidis ATCC 35984 by systematically evaluating growth media, sugar types/concentrations, atmospheric conditions, and incubation periods.Material and Method: S. mutans and S. epidermidis strains were cultured under various conditions, including media (Brain Heart Infusion and Mueller Hinton Broth), sugar types/concentrations (sucrose and glucose), atmospheres (aerobic vs. 5% CO₂), and incubation times (24 and 48 hours). Biofilm production was quantified by standard crystal violet staining in microtiter plates.Result and Discussion: Highest biofilm production for S. mutans occurred in BHI with 2% sucrose, under 5% CO₂, after 24-48 hours. For S. epidermidis, peak biofilm was in MHB with 1% glucose, under aerobic conditions, after 24 hours. These findings highlight the need for species-specific optimization of biofilm-inducing conditions when using standard strains.
Şavluk et al. (2026) studied this question.