Influence of Environmental Factors on Biofilm Formation by Heat-resistant Spore-forming Bacillus cereus TGS11.1

Introduction/Objective This study evaluates the synergistic influence of environmental factors on the biofilm formation capacity of the heat-resistant, spore-forming pathogen Bacillus cereus TGS11.1 within milk-processing contexts. Methods Response Surface Methodology with a Central Composite Design (RSM-CCD) was implemented to quantify the multifactorial effects of temperature (30–70°C), pH (4–8), lactose concentration (2–6%), and incubation time (12–60 h) on biofilm development using optical density measurements and phenotypic motility assays. Results Temperature emerged as the dominant determinant of biofilm formation, with peak productivity and OD570 values occurring at 40°C, correlating with maximum swarming motility and the establishment of a dense three-dimensional (3D) architectural network. Discussion The observed sensitivity of B. cereus to thermal gradients and nutrient availability underscores the need for multifactorial intervention strategies to disrupt the structural resilience of the biofilm matrix during dairy production. Conclusion Mitigating recontamination risks in dairy workflows necessitates the strict avoidance of thermal niches near 40°C to inhibit the optimal physiological conditions for robust biofilm formation by B. cereus TGS11.1.