The amount of sewage sludge generated from wastewater treatment plants globally is unavoidably increasing. In recent years, significant attention has been paid to the biorefinery concept based on the conversion of waste streams to high-value products, material, and energy by microorganisms. However, one of the most significant challenges in the field is the possibility of controlling the microorganisms' pathways in the anaerobic environment. This study investigated two different anaerobic fermentation tests carried out with real waste activated sludge at high organic loading rate (10 g COD L-1d-1) and short hydraulic retention time (HRT) to comprehensively understand whether this configuration enhances extracellular polymeric substance (EPS) and metal solubilisation. The quantity of EPS recovered increased over time, while the chemical oxygen demand to EPS ratio remained in the range 1.31-1.45. Slightly acidic conditions and sludge floc disintegration promoted EPS matrix disruption and release, combined with the solubilisation of organically bound toxic metals, such as As, Be, Cu, Ni, V, and Zn, thereby increasing the overall metal removal efficiency due to the action of hydrolytic microorganisms. Bacteroidetes, Firmicutes, and Chloroflexi were the most abundant phyla observed, indicating that the short HRT imposed on the systems favoured the hydrolytic and acidogenic activity of these taxa.
Keywords: anaerobic hydrolysis; extracellular polymeric substances; high throughput 16S rRNA gene sequencing; microbial communities; toxic metals; waste activated sludge.