A-A. Jones
Duke University,
United States
Keywords: biofilms, P. aeruginosa, standards
Summary:
The Jones Systems for Engaging the Environment Lab builds novel tools to study biofilm dynamics. In this presentation we will discuss a mechanical tool for visualizing Pseudomonas aeruginosa PAO1 interaction with antibiotics. Biofilms are the predominate form of life for bacteria, not the planktonic, or free floating/swimming organisms most people envision when thinking of bacteria. However, most of our tools for studying bacteria and their response to antibiotics are largely planktonic methods. Biofilm responses to antibiotics are quantified and observed using multiple methods, though confocal laser scanning microscopy (CLSM) has many advantages. CLSM can resolve biological and spatial heterogeneity of biofilms in 3D however with limited throughput. We developed a modified microplate peg-lid-based assay optimized for in situ confocal microscopy of all pegs. This modifies the Calgary Biofilm Device, described in ASTM E2799-22, which is a medium-throughput method for testing biofilms but does not permit in situ imaging. We show similar antibiotic challenge recovery via colony formation following the ASTM E2799-22 protocol and in situ imaging. We also show novel quantifiable effects of antibiotics on P. aeruginosa biofilm streamers. We describe preliminary efforts to mathematically model the efficiency of growing biofilms on this device and efforts to integrate this tool with models of antibiotic penetration.