Four hypothetical mechanisms for protection of biofilms against antimicrobials were incorporated into a three-dimensional model of biofilm growth and development. The model integrated processes of substrate utilization, diffusion, growth, cell migration, death, and detachment in a cellular automaton framework. Compared to simulations of unprotected biofilms, each of the protective mechanisms provided some tolerance to antimicrobial action. When the mechanisms were compared to each other, the behaviors of the four protective mechanisms produced distinct shapes of killing curves, nonuniform spatial patterns of survival and cell type distribution, and anticipated susceptibility patterns for dispersed biofilm cells. The differences between the protective mechanisms predicted in these simulations could guide the design of experiments to discriminate antimicrobial tolerance mechanisms in biofilms. Each of the mechanisms could be a plausible avenue of biofilm protection.

Three hypothetical mechanisms of detachment were incorporated into a three-dimensional computer model of biofilm development. The model integrated processes of substrate utilization, substrate diffusion, growth, cell advection, and detachment in a cellular automata framework. The purpose of this investigation was to characterize each of the mechanisms with respect to four criteria: the resulting biofilm structure, the existence of a steady state, the propensity for sloughing events, and the dynamics during starvation. The three detachment mechanisms analyzed represented various physical and biological influences hypothesized to affect biofilm detachment. The first invoked the concept of fluid shear removing biomass that protrudes far above the surface and is therefore subjected to relatively large drag forces. The second pathway linked detachment to changes in the local availability of a nutrient. The third pathway simulated an erosive process in which individual cells are lost from the surface of a biofilm cell cluster. The detachment mechanisms demonstrated diverse behaviors with respect to the four analysis criteria. The height-dependant mechanism produced flat, steady state biofilms that lacked sloughing events. Detachment based on substrate limitation produced significant sloughing events. The resulting biofilm structures included distinct, hollow clusters separated by channels. The erosion mechanism produced neither a non-zero steady state nor sloughing events. A mechanism combining all three-detachment mechanisms produced mushroom-like structures. The dynamics of biofilm decay during starvation were distinct for each detachment mechanism. These results show that detachment is a critical determinant of biofilm structure and of the dynamics of biofilm accumulation and loss.