Spatio-temporal evaluation of anti-biofilm cavitation activity by passive acoustic mapping

Abstract

Bacterial biofilms present a major challenge to achieving effective antibiotic therapy, as these sessile communities of microbes confer protection to bacteria by decreasing antibiotic efficacy. Focused ultrasound can mechanically disrupt biofilms, offering a new ‘drug-free’ antibiotic paradigm. The goal of this work is to validate, quantify, and optimize the role of acoustic cavitation in the biofilm disruption process through spatiotemporal monitoring of cavitation activity. A clinical isolate strain of Staphylococcus aureus from native valve endocarditis was cultured for 72 hours within a flow channel to form a biofilm. A 1.1 MHz spherically focused transducer was used to expose the biofilm from below at a 45° angle. The in situ acoustic field was characterized with a fibre-optic hydrophone. A calibrated 5–11 MHz linear array was placed 25 mm above the biofilm in order to record acoustic emissions during biofilm disruption from which passive acoustic maps of cavitation could be derived. Biofilms were exposed to 4.5 MPa peak rarefactional pressure (derated), 10,000 cycles, at a 1 Hz PRF. Qualitative reduction of biofilms was assessed by live/dead staining with Syto 9/propidium iodide, which was correlated with cavitation activity observed in the passive acoustic maps.