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We took benefit from Atomic Force Microscopy (AFM) in the force spectroscopymode to describe the time evolution-over 24h-of the surface nanotopography andmechanical properties of the strain Staphylococcus aureus 27217 from bacterialadhesion to the first stage of biofilm genesis. In addition, TransmissionElectron Microscopy (TEM) and Scanning Electron Microscopy (SEM) experimentsallowed identifying two types of self-adhering subpopulations (the so-called'bald' and 'hairy' cells) and revealed changes in their relative populationswith the bacterial culture age and the protocol of preparation. We indeedobserved a dramatic evanescing of the 'hairy' subpopulation for samples thatunderwent centrifugation and resuspension processes. When examined by AFM, the'hairy' cell surface resembled to a herringbone structure characterized byupper structural units with lateral dimensions of ~70 nm and a high Young'smodulus value (~ 2.3 MPa), a mean depth of the trough between them of ~15nm anda resulting roughness of ~5nm. By contrast, the 'bald' cells appeared muchsofter (~ 0.35 MPa) with a roughness one order of magnitude lower. We observedtoo the gradual detachment of the herringbone patterns from the 'hairy'bacterial envelope of cell harvested from a 16h old culture and theirprogressive accumulation between the bacteria in the form of globular clusters.The secretion of a soft extracellular polymeric substance was also identifiedthat, in addition to the globular clusters, may contribute to the initiation ofthe biofilm spatial organization.

Direct observation of the cell-wall remodeling in adhering Staphylococcus aureus 27217: an AFM study supported by SEM and TEM