Movement, Replication, and Emigration Rates of Individual Bacteria in a Biofilm

Single-cell behavior within a biofilm was observed over a period of several hours. The observations were converted into quantitative stochastic rules governing the behavior of individual cells within a biofilm. Such a quantitative summary provides not only a concise description of the results but also information helpful when constructing computer models of dynamic biofilm systems. The time to division, emigration, and rate of motility of individual green fluorescent protein labeled Pseudomonas aeruginosa PAO1 cells in a 3-10 microm thick biofilm containing predominantly non-GFP labeled cells were calculated based on images of individual cells collected at 15-min time intervals. The biofilms were grown in flow cells and the images captured with a confocal laser microscope. Cells destined to emigrate are more active than those that remain; the geometric means for velocities in the biofilm are 1.0 microm/h for remaining cells and 1.5 microm/h for emigrating cells. The median time to emigration was 2.0 h. During the experimental observation period, the estimated probability for emigration is 0.44, illustrating that a substantial number of bacteria leave the field of view. Cells emigrate at a median time one-third that of the median time to replication. Specifically, the median time for cells to divide was 6.9 h, and it was estimated that 10% of the cells had a time to division greater than 10 h.