The effects of extrinsic factors on the structural and mechanical properties of Pseudomonas fluorescens biofilms: A combined study of nutrient concentrations and shear conditions

Abstract

The growth of biofilms on surfaces is a complicated process influenced by several environmental factors such as nutrient availability and fluid shear. In this study, combinations of growth conditions were selected for the study of Pseudomonas fluorescens biofilms including as cultivation time (24- or 48 h), nutrient levels (1:1 or 1:10 King B medium), and shear conditions (75 RPM shaking, 0.4 mL min ^-1 or 0.7 mL min ^-1). The use of Confocal Laser Scanning Microscopy (CLSM) determined biofilm structure, while liquid-phase Atomic Force Microscopy (AFM) techniques resolved the mechanical properties of biofilms. Under semi-static conditions, high nutrient environments led to more abundant biofilms with three times higher EPS content compared to biofilms grown under low nutrient conditions. AFM results revealed that biofilms formed under these conditions were less stiff, as shown by their Young's modulus values of 2.35 ± 0.08 kPa, compared to 4.98 ± 0.02 kPa for that of biofilms formed under low nutrient conditions. Under dynamic conditions, however, biofilms exposed to low nutrient conditions and high shear rates led to more developed biofilms compared to other tested dynamic conditions. These biofilms were also found to be significantly more adhesive compared to their counterparts grown at higher nutrient conditions.

Keywords: Atomic force microscopy; Biofilm; Biofilm viscoelastic properties; Confocal laser scanning microscopy; Nutrient concentration; Pseudomonas fluorescens; Shear.