A study of substrate removal by real activated sludge with several synthetic substrates (acetate, ethanol, glutamic acid) and wastewater (raw and filtered) was carried out. Substrate, stored compounds (polyhydroxyalkanoates, PHA and internal carbohydrates), ammonia and oxygen uptake rate (OUR) were analytically determined. Polyhydroxybutyrate (PHB) was stored when the substrate was acetate or ethanol, while no appreciable formation of storage compound was detected using glutamic acid. A low amount of PHB was also formed in tests with raw and filtered wastewater which was probably mainly due to its acetate content. As far as the sum of storage and growth (indirectly estimated through ammonia consumption) did not match the overall solids formation, other unidentified mechanisms of substrate removal were likely to occur (biosorption, accumulation and/or storage of unidentified compounds). ASM3 and two derived models were used in the interpretation of experimental data with reference to synthetic substrates. With reference to synthetic substrates ASM3 can well describe the experimental data only assuming a stored product formation much higher than the analytically detected one, whereas the model that assumes a parallel growth and storage on the substrate can well describe the observed stored product profile only assuming a direct contribution of growth much higher than estimated from ammonia consumption. The model that assumes an accumulation/biosorption stage as first step of substrate removal can better describe the whole experimentally observed behaviour. However as well as in ASM3 this implies that some fraction of removed COD is still unidentified. With reference to real wastewater where the different phenomena were mixed up due to the presence of several substrates, the different models gave similar results.

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