The variation in activated sludge floc composition, flocculation and settling properties was studied at a full scale plant over a 2-year period. A comprehensive set of process parameters was analysed and related to the floc properties to increase the understanding of the factors affecting floc formation. The composition of the activated sludge showed a seasonal change with higher concentrations of extractable extracellular polymeric substances (EPS) during the winter months. The protein content of the total sludge and EPS increased significantly during the winter. This coincided with higher effluent suspended solids concentrations and increased shear sensitivity of the sludge flocs. Only poor correlations between EPS contents and stirred sludge volume index (SSVI) could be observed. High iron concentrations in the sludge due to dosage of iron salt to precipitate phosphorus were found to have a negative impact on the settling and compaction properties of the sludge, whereas it had a positive impact on floc stability. Higher organic loading due to by-passed primary settlers leads to improved settling and compaction properties.