Novel analysis of bacterium-substratum bond maturation measured using a quartz crystal microbalance

Studies in flow displacement systems have shown that the reversibility of bacterial adhesion decreases within seconds to minutes after initial contact of a bacterium with a substratum surface. Atomic force microscopy (AFM) has confirmed that the forces mediating bacterial adhesion increase over a similar time span. The interfacial rearrangements between adhering bacteria and substratum surfaces responsible for this bond maturation have never been studied. Quartz crystal microbalance with dissipation (QCM-D) senses the interfacial region in real time and nondisruptively up to 250 nm from the sensor surface. In this paper, QCM-D is combined with real-time observation of bacterial adhesion in a flow displacement system, in order to analyze resident-time-dependent changes in dissipation. Three different Streptococcus salivarius strains showed a nonlinear relation between total dissipation shift (DeltaD) and number of adhering bacteria, whereas inert and rigid silica particles demonstrated a linear relation between DeltaD and the number of adhering particles. This suggests removal of interfacial water due to residence time dependent deformation of the nonrigid bacterium-substratum interface during bond maturation. Dissipation could be described by an exponentially decaying function, which combined with adhesion data allowed extraction of the dissipation shifts per bacterium upon initial contact (DeltaD(0)), after bond maturation (DeltaD(infinity)), as well as a characteristic time constant (tau(bm)). All bacterial strains showed significant bond maturation within one minute after their arrival at the substratum surface, which was not observed for silica particles. Dissipation analysis at the level of individually adhering bacteria would have been impossible without the simultaneous real-time analysis of bacterial adhesion numbers.