Layered viscoelastic properties of granular biofilms

Granular biofilms are dense spherical complex biological systems composed mainly of multi-microbial cells, water, and extracellular polymeric substances (EPS). They facilitate efficient purification and settling of activated sludge in wastewater treatment processes. The viscoelastic properties of these complex biofilm systems are important characteristics that control their growth and dictate how they respond to hydrodynamic forces and chemical stimuli. However, the viscoelastic properties of granular biofilms are poorly understood. In this paper, we study granular biofilms' viscoelastic properties using optical coherence elastography (OCE), a nondestructive method that integrates optical coherence tomography (OCT) with elastic wave propagation. While quantitative viscoelastic characterization of granular biofilms is challenging due to their heterogeneous properties, we show that elastic waves are suitable for this purpose. First, we employ guided elastic waves in a thin section of a granular biofilm to reveal a two-layered profile for the viscoelastic properties. Next, we utilize circumferential elastic waves that propagate near the surface of a non-sectioned spherical biofilm to quantify the layered system's viscoelastic properties. To the best of our knowledge, this work is the first quantitative study that characterizes the layered viscoelastic properties of granular biofilms. The measurement approach may provide a platform to study the interplay between the viscoelastic properties and other characteristics of granular biofilms such as the complex microbial system, morphology, and oxygen distribution.