Characterization of activated sludge flocs by confocal laser scanning microscopy and image analysis

The purpose of this study was to characterize microbial floc structure and properties under phosphorus (P) limiting and non-limiting regimes. The P-limitation applied to the biomass did not significantly impact on reactor performance in terms of COD removal and MLSS. The composition of EPS was affected by the P-limitation with significantly increased accumulation of carbohydrates, uronic acids and proteins. CLSM and glycoconjugate mapping revealed that the relative abundance of α and β- N -acetylgalactosaminyl/galactopyranosyl and N -acetylglucosaminyl residues was affected by P-limitation, suggesting changes in microbial populations within the floc structure, which in turn could cause the compositional changes of EPS. The image analysis performed on CLSM images indicated that under non-limiting conditions the cell clumps within the floc were significantly smaller as compared to P-limiting conditions. The fractal dimension and porosity under limiting conditions were either significantly higher or lower than under P-rich conditions.