Although the limitation or deficiency of nutrients, such as nitrogen (N) and phosphorus (P), has been one of the frequently reported factors causing filamentous or non-filamentous bulking of activated sludge, the mechanisms are still unclear. In this work, the long-term effects of N and P limitation or deficiency on sludge settleability and bioflocculation characteristics were investigated in six sequencing batch reactors (SBRs) fed with wastewater with different nutrient availability. The sludge volume index (SVI), microbial community structures, intracellular poly-beta-hydroxyalkanoates (PHAs) and extracellular polymeric substances (EPS) were characterised over time. Bulking was not observed in SBRs with N limitation or deficiency, in which SVI remained below 150 mL/g. In contrast, bulking was encountered in those reactors with P deficiency. The occurrence of non-filamentous bulking was associated with a higher carbohydrates fraction and a lower proteins fraction in EPS. In the case of filamentous bulking, SVI correlated negatively with the amount of PHAs. Our experimental data support the hypothesis that the occurrence and/or the type of bulking in activated sludge could be affected by the combination of kinetic selection, microbial storage, as well as the EPS composition. (C) 2014 Elsevier B.V. All rights reserved.