H24A-05
Bacterial Response to Antibiotic Gradients in a Porous Microfluidic Device

Tuesday, 15 December 2015: 17:00
3016 (Moscone West)
Jinzi Deng1, Lang Zhou2, Rajveer Singh2, Yiran Dong2, Charles J Werth3 and Bruce W Fouke2, (1)Institute for Genomic Biology, Urbana, IL, United States, (2)University of Illinois at Urbana Champaign, Urbana, IL, United States, (3)University of Texas at Austin, Department of Civil, Architectural and Environmental Engineering, Austin, TX, United States
Abstract:
Microorganisms in nature have evolved survival strategies to cope with a wide variety of environmental stresses, including gradients in temperature, pH, substrate availability and aqueous chemistry. Microfluidic devices provide a consistently reliable real-time means to quantitatively measure, control and reproduce the dynamic nature of these stresses. As an example, accelerated adaptation from genetic mutations have been observed in E. coli as it responds to gradients of Ciprofloxacin (Zhang et. al. 2011). However, the mechanisms by which bacteria respond to antibiotic gradients, as well as the effect of changes in how the stressor is applied, have not been systematically studied. In this study, newly designed and fabricated microfluidic devices with porous media have been utilized to determine the chemical stress fields that enhance adaptation and thus to test how E. coli bacterial communities adapt to antibiotic stresses. By applying antibiotic and nutrient into inlet channels adjacent to either side of the porous media inoculated with E. coli, a gradient of antibiotic was formed. Hydrogel barriers were selectively photo-polymerized in between of the inlet channels and the porous media to prevent any undesired convection. Hence, chemical solute can only be transported by diffusion, creating a reproducible antibiotic gradient over the porous media. The bacteria were also constrained by the hydrogel boundary barriers from escaping the porous media. Preliminary results suggest that E. coli moves freely with respect to Ciprofloxacin concentrations. In addition, and unexpectedly, the E. coli colonies exhibit a concentric pulsed growth front radiating away from the point of inoculation within the micromodel ecosystem and pulse over the porous media containing antibiotic. The bacteria at the growth front grow into long filaments (up to 100µm) while the bacteria in the inner concentric area are normal size. We hypothesize that the frontier bacteria, which are first exposed to the unfavorable antibiotic concentrations, may initially collapse the gradient and thus reduce the concentration, leaving more favorable growth conditions for the migration of newly replicated bacteria. This approach will also be used to screen other environmental stresses, including nutrient limitation and temperature change.