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GLOMICAVE aims to exploit the information hidden in the existing scientific literature and large-scale -omics dataset for a better understanding of genotype-phenotype relationships. In this deliverable, we will focus on identifying potential polyphosphate accumulating organisms (PAOs) in activated sludge systems, and the prediction of differences in their activity level based on the presence of different carbon sources from metagenomic and metatranscriptomics data. Phosphate is both, a scarce resource and pollutant, which can be found in high volumes in wastewater. To protect the environment, phosphate needs to be removed from wastewater before it goes into the environment after the treatment. Additionally, as an important resource used in fertilizants and not having a natural resource in Europe, it is important to recover phosphate as much as possible. PAOs are present in high abundance in enhanced phosphorus removal (EBPR) wastewater treatment plants (WWTPs), where they play a key role in phosphorus removal and recovery from wastewater. PAOs have the ability to accumulate and store large amounts of inorganic phosphate in their cells as polyphosphate (poly-P). Currently, there are three described PAOs in activated sludge: Candidatus Accumulibacter, Dechloromonas (now Azonexus) and Candidatus Phosphoribacter (former Tetrasphaera) [PSKD22, PSPP21, SPWN22]. In addition to that, it is suggested that there are other unidentified PAOs who also contribute to phosphorus removal and recovery. Recent advantages in -omics enable more in-depth study of PAOs under in situ conditions to better understand their involvement in poly-P accumulation. Generally, under anaerobic conditions, PAOs are thought to
assimilate organic substrates, such as acetate, glucose, amino acids and others to synthesize the storage polymer polyhydroxyalkanoates (PHA), using stored poly-P as an energy source. Under aerobic conditions in the absence of any external organic compounds, the PAOs use PHA for carbon and energy to grow and to re-assimilate phosphate to synthesize poly-P. Captured poly-P is then removed from sewage through surplus sludge discharge. Even though some PAOs and their metabolism are fairly well understood, their in-situ response to different carbon sources is still poorly analyzed and described. Recent advances in -omics enable more in-depth study of PAOs under in situ conditions to better understand their involvement in poly-P accumulation. In this project, we have analyzed time series of microbial communities in full-scale EBPR plants and selected one plant with many PAOs for deeper analysis. A metagenomic analysis was carried out and metagenome assembled genomes (MAGs) were recovered. In total 214 high quality MAGs were recovered representing many abundant species in the systems including the dominant PAOs. Several short-term anaerobic-aerobic lab-scale experiments were carried out with 5 different carbon sources for gene expression studies. In total 60 samples were analyzed to predict PAOs activity within different conditions. The analysis of this data will help further to understand the metabolism of PAOs under different conditions and help with process optimization for optimal phosphate removal and recovery from wastewater.
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