Dynamic Changes in Biofilm Structures under Dynamic Flow Conditions

Quantitative assessment of the responses of biofilm structure to external hydrodynamics is critical for understanding biofilm detachment mechanisms. This study used multidimensional imaging and numerical simulation approaches to elucidate the complex relationships between biofilm detachment and hydrodynamics with Shewanella oneidensis MR-1. By integrating real-time confocal laser scanning microscopy (CLSM) images with image analysis tools, the three-dimensional structural changes occurring in thin MR-1 biofilms (<10 μm) under hydrodynamic treatment at a flow velocity of 0.42 × 10^-3 to 3.3 × 10^-3 m/s in the laminar flow regime were visualized in situ and quantified with single-cell resolution. Analyses of the imaging results revealed high spatial heterogeneity in the degree and intensity of biofilm detachment. Spots with thick and rough biofilm surfaces or high flow rates had high detachment rates, indicating that local biofilm morphology, including thickness and roughness, and hydrodynamic flow conditions collectively controlled the detachment rate. Numerical simulations revealed a significant correlation between local detachment events and the shear stress induced by hydraulic flow at the three-dimensional level. Compared to the even or thin biofilm, a thick or rough structure might induce a 2-fold increase in shear stress over local biofilm surfaces at a microscale dimension. The results provide quantitative and microscopic insights into biofilm detachment processes in subsurface environments, especially in domains under dynamic flow conditions, such as those in hyporheic zones. The relationship between biofilm detachment and hydrodynamics and biofilm structural factors can be integrated into reactive transport models used to describe microbial growth and transport in porous media. IMPORTANCE Detachment is an important process determining the structure and function of bacterial biofilm, which has significant implications for biogeochemical cycling of elements, biofilm application, and infection control in clinical settings. Quantifying the responses of biofilm structure to hydrodynamics is crucial for understanding biofilm detachment mechanisms in aquatic environments. In this work, the spatial and temporal changes occurring in biofilm structures in response to different hydrodynamic conditions were studied by using flow cell reactors. We established the quantitative relationships among detachment, biofilm morphology, and shear stress induced by changes in hydrodynamic conditions. This work provides a quantitative understanding of the complex relationship between biofilm detachment and hydrodynamics in natural environments.