An expanding bacterial colony forms a depletion zone with growing droplets

Abstract

Many species of bacteria have developed means to spread on solid surfaces. This study focuses on the expansion of Pseudomonas aeruginosa on an agar gel surface. We report the occurrence and spread of a depletion zone, where the layer of bacteria on the agar becoming thinner. The depletion zone occurs within an expanded colony under conditions of minimal water evaporation. It is colocalized with a higher concentration of rhamnolipids, the biosurfactants that are produced by the bacteria and accumulate in the older region of the colony. With continued growth in bacterial population, dense droplets occur and coalesce in the depletion zone, displaying remarkable fluid dynamic behavior. Whereas expansion of a central depletion zone requires activities of live bacteria, new zones can be seeded by adding rhamnolipids. These depletion zones due to the added surfactants expand quickly, even on plates covered by bacteria that have been killed by ultraviolet light. We propose a model to account for the observed properties, taking into consideration bacterial growth and secretion, osmotic swelling, fluid volume expansion, cell-cell interaction, and interfacial fluid dynamics involving Marangoni flow. SignificanceBacterial growth and pattern formation have strong bearing on their biological functions, such as their spread and accumulation, biofilm growth & its effects on infection and antibiotic resistance. The bacterial species of this study, Pseudomonas aeruginosa, is a human pathogen responsible for frequent infections in wounds, airways, and urinary tract, particularly when involving the use of catheters. The findings of this study and the mechanisms we propose offer new insights on the important behaviors of bacterial collective motility, pattern dynamics, and biofilm growth.