Optimization of Biofilm Formation: Influence of Temperature, pH, and Incubation Time in a Factorial Experimental Framework

This study systematically evaluated the influence of temperature, pH, and incubation time on biofilm formation using a full-factorial experimental design and response surface methodology (RSM). The results demonstrated that all three independent variables significantly impacted biofilm growth (p < 0.0001, ANOVA), with optimal biofilm formation occurring at 37°C, pH 7.0, and 36 hours of incubation. Deviations from these optimal conditions led to substantial reductions in biofilm biomass, with acidic (pH 5.0) and alkaline (pH 9.0) conditions reducing biofilm formation by 40% and 35%, respectively, and extreme temperatures (25°C and 45°C) inhibiting biofilm formation by more than 30% and 50%, respectively. The predictive quadratic regression model developed in this study exhibited high accuracy (R² = 98.3%), with a prediction error of less than 2% when validated experimentally. This confirms the robustness and reliability of the optimization framework in biofilm-related studies. The findings offer valuable insights into microbial adhesion dynamics, providing a foundation for biofilm management strategies in healthcare, biotechnology, and industrial applications. The insights gained from this research provide a strong foundation for biofilm management and optimization strategies across various fields, including medical microbiology, industrial biotechnology, and environmental sciences. By integrating advanced experimental approaches, computational modeling, and interdisciplinary collaboration, future research can pave the way for innovative biofilm control strategies and biofilm-enhanced biotechnological applications.

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