Understanding the microbial ecology and ecophysiology of enhanced biological phosphorus removal processes through metabolic modelling and experimental studies

Dissertation to obtain the degree of Master in Chemical and Biochemical Engineering

The enhanced biological phosphorus removal (EBPR) process in activated sludge systems has become a widely applied wastewater treatment technology to control eutrophication. The success of this process relies on the sludge enrichment with polyphosphate accumulating organisms (PAOs), while one of the main causes for its failure is due to microbial competition between PAOs and another group of organisms known as the glycogen accumulating organisms (GAOs). The microbial ecology and ecophysiology of these two groups have been investigated through metabolic modelling and experimental studies in order to provide a better understanding of EBPR systems. This thesis focuses on researching the P removal efficiency and metabolic behaviour of an enriched culture containing two PAOs: Tetrasphaera-related organisms and Accumulibacter, which were acclimatized with casamino acids as sole carbon source in a sequencing batch reactor (SBR). Both organisms were identified through fluorescence in situ hybridization (FISH), and this culture demonstrated anaerobic P release, glycogen hydrolysis, a very low poly--hydroxyalkanoates (PHA) synthesis and high casamino acids uptake; followed by aerobic P uptake, glycogen formation and a very low PHA oxidation. Different carbon sources (glucose, acetate, propionate, glutamate, aspartate, glycine and casamino acids) were studied through batch tests inoculated with sludge from the main SBR. Through experimental data, it was suggested that Accumulibacter were responsible for the uptake of volatile fatty acids (VFAs), and Tetrasphaera-related organisms were likely responsible for both glucose and amino acids uptake. This thesis also focuses on the development of a model that combines a PAO-GAO metabolic model with activated sludge model no. 2d (ASM2d) in collaboration with Hydromantis Environmental Software Solutions, Inc.. The combined model was implemented in the GPS-X software and will provide a new and advanced platform for wastewater treatment modelling, which will be available to practitioners.