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Insufficient influent carbon source affects the performance and stability of the biological nutrient removal (BNR) process in wastewater treatment plants (WWTPs). To solve this problem, a side-stream activated sludge hydrolysis (SSH) process that utilizes the internal carbon source through hydrolysis and fermentation of return activated sludge (RAS) was proposed as an alternative. In this study, two laboratory-scale pilot reactors were run to compare the pollutant removal performance in SSH and conventional anaerobic/anoxic/aerobic (A2/O) processes at various influent food-to-microorganism (F/M) ratios (0.07–0.71). When the average F/M ratio increased to > 0.35, the chemical oxygen demand (COD), ammonia (NH4+-N), total nitrogen (TN), and total phosphorus (TP) removal efficiencies in SSH reactor reached ~ 94%, > 95%, > 90%, and > 85%, respectively. Under the same influent conditions, the TN removal in SSH reactor was 23% higher than that in A2/O reactor, which is related to the additional carbon generated via RAS fermentation and mitigated substrate competition among various heterotrophic organisms. A relatively high polyphosphate accumulating organism (PAO) activity was also observed in SSH reactor. Meanwhile, the relative abundances of denitrifiers (e.g., Thiothrix, Azospira, and Dechloromonas) and PAOs (i.e., Accumulibacter, Tetrasphaera, and Dechloromonas) were higher in SSH reactor than those in A2/O reactor. These results will provide a better understanding of the mechanisms involved in SSH process, leading to more effective nutrient removal for treating wastewater with limited carbon sources. [ABSTRACT FROM AUTHOR]

Dissolved oxygen (DO) plays an important role in the performance of biological wastewater treatment systems. This study investigated the effect of the DO concentration on nutrient removal performance and microbial community structure in side-stream activated sludge hydrolysis (SSH) and conventional anaerobic/anoxic/aerobic (A2O) processes. The results showed that the change in DO had little effect on the removal performance of chemical oxygen demand (COD), and the removal efficiencies were about 90% for both reactors. Compared with the high DO level (4.1–6.9 mg/L), the A2O and SSH reactors had better nitrogen removal performance at low (0.5–2.2 mg/L) and moderate (2.2–3.9 mg/L) DO levels, with ammonia (NH4+-N) removal efficiencies of 88–89% and 89–91%, respectively, and total nitrogen (TN) removal efficiencies of 74–76% and 75–81%, respectively. Directly reducing the DO concentration from high to low reduced the phosphate removal efficiencies of the A2O and SSH reactors from 80.2% and 86.2% to 63.1% and 70.6%, respectively, while re-elevating the DO concentration to moderate levels significantly improved the phosphate removal efficiencies to 94.6% and 96.0%, respectively. Compared to the A2O reactor, the SSH reactor had more stable and better nutrient removal performance under different DO conditions, partly due to the additional carbon sources produced through the sludge fermentation in the side-stream reactor. The decrease in the DO concentration resulted in a decrease in the relative abundance of Acinetobacter but an increase in the relative abundance of Competibacter, potentially leading to the deterioration in phosphorus removal. [ABSTRACT FROM AUTHOR]

Polyphosphate accumulating organisms (PAOs) belong mostly to Proteobacteria and Actinobacteria and are quite divergent. Under aerobic conditions, they accumulate intracellular polyphosphate (polyP), while they typically synthesize polyhydroxyalkanoates (PHAs) under anaerobic conditions. Many ecological, physiological, and genomic analyses have been performed with proteobacterial PAOs, but few with actinobacterial PAOs. In this study, the whole genome sequence of an actinobacterial PAO, Microlunatus phosphovorus NM-1T (NBRC 101784T), was determined. The number of genes for polyP metabolism was greater in M. phosphovorus than in other actinobacteria; it possesses genes for four polyP kinases (ppks), two polyP-dependent glucokinases (ppgks), and three phosphate transporters (pits). In contrast, it harbours only a single ppx gene for exopolyphosphatase, although two copies of ppx are generally present in other actinobacteria. Furthermore, M. phosphovorus lacks the phaABC genes for PHA synthesis and the actP gene encoding an acetate/H+ symporter, both of which play crucial roles in anaerobic PHA accumulation in proteobacterial PAOs. Thus, while the general features of M. phosphovorus regarding aerobic polyP accumulation are similar to those of proteobacterial PAOs, its anaerobic polyP use and PHA synthesis appear to be different. [ABSTRACT FROM PUBLISHER]

Protein expression is a direct reflection of specific microbial activities in any ecosystem. In order to assess protein expression in mixed microbial communities, the feasibility of applying proteomic techniques to activated sludge samples has recently been demonstrated. We report the application of metaproteomics to two activated sludges from a laboratory-scale sequencing batch reactor with dissimilar phosphorus removal performances. Fluorescence in situ hybridization (FISH) revealed that the sludge with good enhanced biological phosphorus removal performance (EBPR) was dominated by Betaproteobacteria (65% of EUBMIX binding cells) and gave positive signals for the Rhodocyclus-type PAO specific probe (59%). The non-EBPR sludge was dominated by tetrad-forming Alphaprofeobacferia (75%). With regard to the proteomic investigation, 630 individual protein spots were matched across the replicate groups of the anaerobic and aerobic phases of the EBPR sludge with 9.4% of all spots being statistically different between the two phases. The non-EBPR metaproteomic maps exhibited 590 matched spots with 14.7% statistical differences between the two phases. Overall, the non-EBPR sludge expressed around 30% more significant differences than the EBPR sludge. The comparison of protein expression in the two sludges showed that their metaproteomes were substantially different and this was reflected in their microbial community structures and metabolic transformations. [ABSTRACT FROM AUTHOR]

The effectiveness of enhanced biological phosphorus removal (EBPR) systems is directly affected by the competition of polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs). This study investigated the short-term effects of carbon source on PAO and GAO performance. The tests were designed to clearly determine the impact of volatile fatty acid (VFA) composition on the performance of two types of biomass, one enriched for PAOs and the other for GAOs. The two populations were enriched in separate reactors using identical operating conditions and very similar influent compositions with acetate as the sole carbon source. The only difference was that a very low level of phosphorus was present in the influent to the GAO reactor. The abundance of PAOs and GAOs was quantified using fluorescence in-situ hybridisation. The results clearly show that there are some very distinctive differences between PAOs and GAOs in their ability to utilise different carbon substrates. While both are able to take up acetate rapidly and completely, the GAOs are far slower at consuming propionate than the PAOs during short-term substrate changes. This provides a potentially highly valuable avenue to influence the competition between PAOs and GAOs. Other VFAs studied seem to be less usable in the short term by both PAOs and GAOs, as indicated by their much lower uptake rates. [ABSTRACT FROM AUTHOR]

This study investigated the effects of different carbon sources on enriched Accumulibacter PAO cultures at high temperature (30 °C) and compared the carbon transformation with low temperature (20 °C) cases reported in literature, revealing several key metabolic differences. While PAOs seemed to prefer propionate anaerobically as compared to other VFAs at high temperature, high aerobic glycogen replenishment was realized with propionate as the anaerobic carbon source, a trait not previously observed at low temperatures. Therefore, it was found that propionate is not correlated with high P removal by Accumulibacter PAO at high temperatures. A combined substrate of acetate, propionate and perhaps butyrate seemed to be a better carbon source combination, since the total VFA uptake rate increased by up to 46%, and this increased the aerobic P-removal efficiency by up to 38.4% and reduced the glycogen recovery by more than 63% compared to the use of only propionate as substrate. This study improves our understanding of how to stimulate successful EBPR operation in warm climates by augmenting the P removal performance of PAOs. • High temperature affects PAO's kinetics but not its anaerobic stoichiometry. • PAO could utilize butyrate and iso-butyrate, hardly used valerate and iso-valerate. • Propionate as sole carbon source causes high aerobic glycogen replenishment. • A combination of acetate, propionate and perhaps butyrate may be a better for EBPR. • Multi-cycle operation may be crucial for PAOs outcompete GAOs at high temperature. [ABSTRACT FROM AUTHOR]