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The present study evaluated different sludge-reduction mechanisms in the oxic-settling-anaerobic (OSA) process in terms of their effects on methane productivity by anaerobic digestion of sewage sludge. Two different layouts were investigated for the sludge return from an anaerobic side-stream reactor (ASSR) to the anoxic (scheme A) or the aerobic (scheme B) reactor of a pre-denitrification plant. Biochemical methane-potential (BMP) assays performed on the excess sludge revealed that scheme A promoted an overall increase of methane production in the OSA (20 mLCH4 gVSS−1d−1, +19%), although compared with a control CAS plant a significant decrease in the excess sludge production (31%) was obtained. Operating conditions in scheme A caused the occurrence of cell lysis and EPS hydrolysis, thereby increasing the biodegradability of sludge. In contrast, scheme B favoured the occurrence of uncoupling and a maintenance metabolism that did not involve sludge hydrolysis. Consequently, despite a higher reduction of excess sludge (82%), a significant decrease in methane productivity in the OSA (4 mLCH4 gVSS−1d−1, −41%) was observed. Based on the results, implementing the OSA process may allow high levels of methane production by anaerobic digestion to be maintained if specific sludge-reduction mechanisms are triggered in the waterline, also raising the possibility of co-digestion with other feedstocks.

The recovery of biopolymers from sewage sludge could be a crucial step in implementing circular economy principles in wastewater treatment plants (WWTP). In this frame, the present study was aimed at evaluating the simultaneous production of polyhydroxyalkanoates (PHA) and extracellular polymeric substances (EPS) obtainable from the treatment of agro-industrial wastewater. Two biological enrichment systems, aerobic granular sludge (AGS) and a conventional activated sludge operating as a sequencing batch reactor (SBR), were monitored for 204 and 186 days, respectively. The maximum biopolymers accumulation capacity was close to 0.60 mgPHA-EPS gVSS−1 in the AGS when operating at 3 kgCODm−3d−1, whereas in the SBR, it was about half (0.35 mgPHA-EPS gVSS−1). Biopolymers extracted from the AGS were mainly constituted by EPS (>70%), whose percentage increased up to 95% with the OLR applied in the enrichment reactor. In contrast, SBR enabled obtaining a higher PHA production (50% of the biopolymers). Results suggested that organic carbon was mainly channeled toward metabolic pathways for extracellular storing in AGS, likely due to metabolic stressors (e.g., hydraulic selection pressure, shear forces) applied for promoting aerobic granulation.

Fouling is considered one of the main drawbacks of membrane bioreactor (MBR) technology. Among the main fouling agents, extracellular polymeric substances (EPS) are considered one of the most impactful since they cause the decrease of sludge filterability and decline of membrane flux in the long term. The present study investigated a biological strategy to reduce the membrane-fouling tendency in MBR systems. This consisted of seeding the reactor with activated sludge enriched in microorganisms with polyhydroxyalkanoate (PHA) storage ability and by imposing proper operating conditions to drive the carbon toward intracellular (PHA) rather than extracellular (EPS) accumulation. For that purpose, an MBR lab-scale plant was operated for 175 days, divided into four periods (1–4) according to different food to microorganisms’ ratios (F/M) (0.80 kg COD kg TSS−1 d−1 (Period 1), 0.13 kg COD kg TSS−1 d−1 (Period 2), 0.28 kg COD kg TSS−1 d−1 (Period 3), and 0.38 kg COD kg TSS−1 d−1 (Period 4)). The application of the feast/famine strategy favored the accumulation of intracellular polymers by bacteria. The increase of the PHA accumulation inside the cells corresponded to the decrease of EPS and an F/M of 0.40–0.50 kg COD kg TSS−1 d−1 was found as optimum to maximize the PHA production, while minimizing EPS. The lowest EPS content in the sludge (18% of total suspended solids) that corresponded to the maximum content of PHA (9.3%) was found in Period 4 and determined significant mitigation of the fouling rate, whose value was close to 0.10 × 1011 m−1 h−1. Thus, by imposing proper operating conditions, it was possible to drive the organic matter toward PHA accumulation. Moreover, a lower EPS content corresponded to a decrease in the irreversible fouling mechanism, which would imply a lower frequency of the extraordinary cleaning operations. This study highlighted the possibility of obtaining a double benefit by applying an MBR system in the frame of wastewater valorization: minimizing the fouling tendency of the membrane and recovery precursors of bioplastics from wastewater in line with the circular economy model.

The production of polyhydroxyalkanoates (PHA) using industrial wastewaters as feedstocks is a current and challenging topic. This study investigated the production of biopolymers by a mixed microbial culture under different OLRs equal to 1 kgCOD m-3d-1 (Period 1), 2 kgCOD m-3d-1 (Period 2) and 3 kgCOD m-3d-1 (Period 3). The maximum PHA content was achieved in Period 2 (0.38 gPHA gTSS-1), whereas lower values were obtained in Period 1 (0.13 gPHA gTSS-1) and Period 3 (0.26 gPHA gTSS-1). Overall, the maximum PHA productivity resulted equal to 0.08 gPHA L-1h-1 (P2), 0.05 gPHA L-1h-1 (P1) and 0.04 gPHA L-1h-1 (P3), respectively. The molecular weight of the PHA increased from Period 1 (250 kDa) to Period 2 (417 KDa) and Period 3 (463 KDa), although resulting in a slight decrease of crystallinity degree. Microbial community analysis, revealed a reduction in bacterial diversity and a progressive shift of the microbial community with the increasing OLR. Alpha-diversity indexes based on Operational Taxonomic Units (OTUs) at 99% identity revealed higher species richness (Taxa (S) 280) and diversity (Shannon (H) 4,06) in Period 1, whereas Period 3 was characterized by reduced richness and diversity and higher dominance (Taxa (S) 133, Shannon (H) 2,40). Based on the results obtained, it was pointed out that the OLR variation determined significant effects on the process performances, as well as on the productivity and quality of the biopolymers. This means that OLR is a key control parameter to maximize the PHA production and control the physical-chemical characteristics of the polymers.