In this study, the production of polyhydroxyalkanoates (PHAs) from waste activated sludge (WAS) was evaluated. PHAs were produced from fermented WAS pretreated via high-pressure thermal hydrolysis, a stream characterised by high levels of nutrients (approximately 3.5 g N L(-1) and 0.5 g P L(-1)) and soluble organics. PHA-storing organisms were successfully enriched at high organic loading rates (6 g COD(sol) L(-1) d(-1)) under aerobic dynamic feeding in sequencing batch reactors at a sludge retention time of 6 d with a short feast length less than 20% of the cycle, and a maximum substrate concentration during feast of 1 g COD(VFA) L(-1). The biomass enrichment, characterised by a decrease in species evenness based on Lorenz curves, provided a biomass that accumulated 25% PHA on a dry-biomass basis with yields on VFA of 0.4 Cmol Cmol(-1) in batch tests. The PHA consisted of ∼70 mol% 3-hydroxybutyrate and ∼30 mol% 3-hydroxyvalerate, and presented high thermal stability (T(d) = 283-287 °C) and a molecular mass ranging from 0.7 to 1.0 × 10(6) g mol(-1). Overall PHA storage was comparable to that achieved with other complex substrates; however, lower PHA storage rates (0.04-0.05 Cmol PHA(-1) Cmol X(-1) h(-1)) and productivities (3-4 Cmol PHA L(-1) h(-1)) were probably associated with a biomass-growth and high-respiration response induced by high levels of non-VFA organics (40-50% of COD(sol) in feed) and nutrients. PHA production is feasible from pretreated WAS, but the enrichment and accumulation process require further optimisation. A milder WAS pretreatment yielding lower levels of non-VFA organics and readily available nutrients may be more amenable for improved performance.