Effect of extraction method on the structure and bioactivity of polysaccharides from activated sludge.

Resource recovery is a pivotal facet of waste activated sludge treatment, particularly within the framework of carbon neutrality and the circular economy. Polysaccharides are emerging as a valuable resource from waste activated sludge, and the choice of extraction method affects the properties of the polysaccharides, which is of utmost importance for subsequent application. This investigation examined the effects of six extraction methods (i.e., acidic, alkaline, ultrasonication, hot-water, microwave, and electric treatments) on the yield, chemical composition, structural characteristics, and bioactivities of polysaccharides extracted from sludge. For each extraction method, two operational parameters, namely the treatment time and strength (e.g., the acid and alkali concentration), were initially optimized in terms of the polysaccharide yield. The polysaccharide yield varied from 1.03 ± 0.12 % to 5.34 ± 0.10 % adopting the extraction methods under optimized conditions, and the alkaline extraction method had the highest yield of polysaccharides with a treatment time of 120 min and NaOH concentration of 1 %. At least one polysaccharide fraction was successfully purified from the crude polysaccharide of each extraction method. The compositions and structures of these fractions, including carbohydrate, protein, sulfate, uronic acid contents, and monosaccharide compositions, were determined. Carbohydrate was the dominant component, with the hot-water-2 fraction having the highest carbohydrate content (77.90 % ± 2.02 %). Monosaccharides in the polysaccharides were measured, with mannose, rhamnose, glucose, and xylose being found in all fractions, whereas ribose was exclusively found in the acid-1 fraction. The molecular weights of these fractions ranged between 1.60 × 104 Da and 7.11 × 106 Da. Furthermore, the bioactivities of the polysaccharides, encompassing five anti-oxidant and three anti-coagulant properties, were assessed, with the ultrasonication-1 fraction having superior performance in seven of the assays. Finally, the association among the fractions in terms of composition and bioactivity was assessed adopting cluster analysis and regression methods. The findings underscore the effect of the extraction method on the properties of polysaccharides extracted from sludge, thereby providing valuable insights for the prospective applications of polysaccharides.

Sulphate reduction, mixed sulphide- and thiosulphate-driven Autotrophic denitrification, NItrification, and Anammox (SANIA) integrated process for sustainable wastewater treatment.

This study proposes the Sulphate reduction, mixed sulphide- and thiosulphate-driven Autotrophic denitrification, Nitrification, and Anammox integrated (SANIA) process for sustainable treatment of mainstream wastewater after organics capture. Three moving-bed biofilm reactors (MBBRs) were applied for developing sulphate reduction (SR), mixed sulphide- and thiosulphate-driven partial denitrification and Anammox (MSPDA), and NItrification (N), respectively. Typical mainstream wastewater after organics capture (e.g., chemically enhanced primary treatment, CEPT) was synthesized with chemical oxygen demand (COD) of 110 mg/L, sulphate of 50 mg S/L, ammonium of 30 mgN/L. The feasibility of SANIA was investigated with mimic nitrifying effluent supplied in MSPDA-MBBR (Period I), followed by the examination of the applicability of SANIA process with N-MBBR integrated (Period II), under moderate temperatures (25-27 â„ƒ). In Period I, SANIA process was established with both SR- and MSPDA-MBBR continuously operated for over 300 days (no Anammox biomass inoculation). Specifically, in MSPDA-MBBR, high rates of denitratation (2.7 gN/(m2·d)) and Anammox (2.8 gN/(m2·d)) were achieved with Anammox contributing to 81 % of the total inorganic nitrogen removal. In Period II, the integrated SANIA system was continuously operated for over 130 days, achieving up to 90 % of COD, 93 % of ammonium, and 61 % of total inorganic nitrogen (TIN) removal, with effluent concentrations lower than 10 mg COD/L, 3 mg NH4+-N/L, and 13 mg TIN-N/L. The implementation of SANIA can ultimately reduce 75 % and 40 % of organics and aeration energy for biological nitrogen removal. Considering the combination of SANIA with CEPT for carbon capture and sludge digestion/incineration for energy recovery, the new integrated wastewater technology can be a promising strategy for sustainable wastewater treatment.

Coupling methane and bioactive polysaccharide recovery from wasted activated sludge: A sustainable strategy for sludge treatment.

Bioactive polysaccharides (PSs) are valuable resources that can be extracted from waste activated sludge (WAS). The PS extraction process causes cell lysis that may enhance hydrolytic processes during anaerobic digestion (AD) and thus increase the methane production. Thus, coupling PSs and methane recovery from WAS could be an efficient and sustainable sludge treatment. In present study, we comprehensively evaluated this novel process from the efficiencies of different coupling strategies, properties of the extracted PSs, and environmental impacts. The results showed that when the PS extraction was before AD, it produced 76.03 ± 2.00 mL of methane per gram of volatile solids (VS) and afforded a PS yield of 6.3 ± 0.09% (w:w), with a PS sulfate content of 13.15% ± 0.06%. In contrast, when PS extraction was after AD, the methane production decreased to 58.14 ± 0.99 mL of methane per gram of VS and afforded a PS yield of 5.67% ± 0.18% (w:w) in VS, with a PS sulfate content of 2.60% ± 0.04%. When there were two PS extractions before and after AD, the methane production, PS yield and sulfate content were 76.03 ± 2.00 mL of methane per gram of VS, 11.54 ± 0.62% and 8.35 ± 0.12%, respectively. Then, the bioactivity of the extracted PSs was assessed by one anti-inflammation assay and three anti-oxidation assays, and statistical analysis revealed that these four bioactivities of PSs were influenced by their sulfate content, protein content and monosaccharide composition, especially the ratios of arabinose and rhamnose. Furthermore, the environmental impact analysis shows that S1 was the best in five environmental indicators compared with other three non-coupled processes. These findings suggest that the coupling PSs and methane recovery process should be further explored to determine its potential for large-scale sludge treatment.

Achieving superior nitrogen removal in an air-lifting internal circulating reactor for municipal wastewater treatment: Performance, kinetic analysis, and microbial pathways.

Anticipated growth in living standards has accentuated higher requirements for effluent quality from municipal wastewater treatment. In this study, an air-lifting internal circulating reactor with a high internal circulation ratio (36:1) was established to treat municipal wastewater with a long-term operation. In the bioreactor, the average effluent chemical oxygen demand, total nitrogen, and ammonium nitrogen could be 13.1, 5.7, and lower than 1 mg/L, respectively. Further analysis of nitrogen removal showed that traditional nitrification and denitrification, simultaneous nitrification and denitrification (SND), and nitrogen assimilation accounted for 27.4 %, 68.7 %, and 3.9 % respectively. The proportion of aerobic bacteria (Saprospiraceae) and facultative bacteria (Comamonadaceae) were significantly increased, indicating a higher capacity for organic degradation in the reactor. The relative abundance of denitrifying bacteria and bacterial groups with SND (Comamonadaceae) increased. These results suggested the air-lifting internal circulating reactor could be a viable and efficient option for superior nitrogen removal in wastewater treatment.

Coupling sulfur-based denitrification with anammox for effective and stable nitrogen removal: A review.

Anoxic ammonium oxidation (anammox) is an energy-efficient nitrogen removal process for wastewater treatment. However, the unstable nitrite supply and residual nitrate in the anammox process have limited its wide application. Recent studies have proven coupling of sulfur-based denitrification with anammox (SDA) can achieve an effective nitrogen removal, owing to stable provision of substrate nitrite from the sulfur-based denitrification, thus making its process control more efficient in comparison with that of partial nitrification and anammox process. Meanwhile, the anammox-produced nitrate can be eliminated through sulfur-based denitrification, thereby enhancing SDA's overall nitrogen removal efficiency. Nonetheless, this process is governed by a complex microbial system that involves both complicated sulfur and nitrogen metabolisms as well as multiple interactions among sulfur-oxidising bacteria and anammox bacteria. A comprehensive understanding of the principles of the SDA process is the key to facilitating the development and application of this novel process. Hence, this review is conducted to systematically summarise various findings on the SDA process, including its associated biochemistry, biokinetic reactions, reactor performance, and application. The dominant functional bacteria and microbial interactions in the SDA process are further discussed. Finally, the advantages, challenges, and future research perspectives of SDA are outlined. Overall, this work gives an in-depth insight into the coupling mechanism of SDA and its potential application in biological nitrogen removal.

Effects of the aeration mode on nitrogen removal in a compact constructed rapid infiltration system for advanced wastewater treatment.

The configuration and the effective operation of constructed rapid infiltration (CRI) systems are of significance for advanced wastewater treatment. In this study, a novel CRI system was developed with a compact structure consisting of two stages, i.e., oxic and anoxic stages. The CRI system was continuously operated for about 140 days under different aeration modes, i.e., tidal flow, continuous aeration, and intermittent aeration. Nitrogen removal was not desirable with tidal flow due to the insufficient oxygen supply in the oxic stage for nitrification, while continuous aeration could achieve good performance for chemical oxygen demand (COD), ammonium, total nitrogen (TN), and total phosphorus (TP) removal. By comparison, the CRI system operated with intermittent aeration was more favorable due to the effective removal ability for pollutants and relatively lower energy demand. The microbial community analysis revealed that Proteobacteria was the dominant phylum in both oxic and anoxic stages of the developed CRI system. Functional microbial groups (Plasticicumulans, Pseudomonas, and Nitrospira in the oxic stage; Thauera, Candidatus_Competibacter, and Dechloromonas in the anoxic stage) were identified for the mediation of carbon, nitrogen, and phosphorus in the system. This study evaluated the feasibility and the optimal aeration mode of the developed CRI system for advanced wastewater treatment, which could satisfy the requirement for the high standard of effluent quality.

Development of a kinetic model to evaluate thiosulfate-driven denitrification and anammox (TDDA) process.

Recently, the integration of sulfur-driven denitrification and anammox process has been extensively studied as a promising alternative nitrogen removal technology. Most of these studies investigated the process feasibility and monitored the community dynamics. However, an in-depth understanding of this new sulfur-nitrogen cycle bioprocess based on mathematical modeling and elucidation of complex interactions among different microorganisms has not yet been achieved. To fill this gap, we developed a kinetic model (with 7 bioprocesses, 12 variables, and 19 parameters) to assess the sulfur(thiosulfate)-driven denitrification and anammox (TDDA) process in a single reactor. The parameters used in this process were separately estimated by fitting the data obtained from the experiments. Then, the model was further validated under different conditions, and the results demonstrated that the developed model could describe the dynamic behaviors of nitrogen and sulfur conversions in the TDDA system. The newly developed branched thiosulfate oxidation model was also verified by conducting a metagenomics analysis. Using the developed model, we i) examined the interactions between sulfur-oxidizing bacteria and anammox bacteria at steady-state conditions with varying substrates to demonstrate the reliability of TDDA, and ii) evaluated the feasibility and operation of the TDDA process in terms of practical implementation. Our results will benefit further exploration of the significance of this novel S-N cycle bioprocess and guide its future applications.

Exploration and verification of the feasibility of sulfide-driven partial denitrification coupled with anammox for wastewater treatment.

Anaerobic ammonia oxidation (anammox) is a well-developed biotechnology for treating high-strength ammonium wastewaters. Recently, partial denitrification has been considered as an alternative to supply anammox with the required nitrite. In this study, a process of sulfide-driven partial denitrification and anammox (SPDA) was developed and operated continuously in an upflow anaerobic sludge blanket (UASB) reactor for 392 days. This reactor was fed with synthetic wastewater containing 100 mgN/L nitrate, 80 mgN/L ammonium and 20-80 mgS/L sulfide. After 160 days of operation, the reactor reached stable performance, and the nitrogen removal efficiency and rate were maintained at 80% and 0.29 kgN/(m³â€¢d), respectively. The estimated nitrogen removal via anammox and sulfide-driven denitrification were 87.2% and 12.8%. Additional batch experiments were conducted to investigate the effects of sulfide on anammox and the mechanisms of nitrogen removal in the SPDA system. The following results were obtained: (1) sulfide had an inhibitory effect on the specific anammox activity with IC50 of 9.7 mgS-H2S/L. (2) The rapid oxidation of sulfide by sulfur-oxidizing bacteria (SOB) could relieve the toxic effects of sulfide on the anammox in the SPDA system. (3) Sulfide bio-oxidation was a two-step reaction with biologically produced elemental sulfur (BPS0) as the intermediate, and the second step using BPS0 as the electron donor, can efficiently produce nitrite via partial denitrification (NO3- → NO2-) as a supply for anammox. Finally, a high-throughput sequencing analysis identified Thiobacillus and Sulfurimonas as the dominant genera of SOB in the SPDA system, and Candidatus Kuenenia as the dominant anammox bacteria. Overall, this research gives the foundation for the practical application of sulfide-driven partial denitrification and anammox process in the future.

Identifying the mechanisms of sludge reduction in the sulfidogenic oxic-settling anaerobic (SOSA) process: Side-stream sulfidogenesis-intensified sludge decay and mainstream extended aeration.

An energy-/cost-efficient and environment-friendly in-situ sludge reduction process, called the sulfidogenic oxic-settling anaerobic (SOSA) was developed recently. However, the underpinning mechanism of sludge reduction by the SOSA process remains elusive. This paper investigated the possible mechanisms of sludge reduction through biomass cultivation in three lab-scale experimental systems: one anoxic-oxic CAS process with a long sludge retention time (SRT) and extended aeration (EAO) process, and two EAO-based in-situ sludge reduction processes, i.e., the conventional oxic-settling anaerobic (COSA) process and the new SOSA process. These three comparative biosystems were operated with identical influent and reactor configurations as well as the same biomass concentrations and SRTs (approximately 5 g/L and 46 days, respectively), and the sludge interchange ratios (between the CAS and side-stream reactors) in COSA and SOSA were both 10% per day. Three systems all achieved high organic (>93%) and total nitrogen (TN) (>74%) removal efficiencies. SOSA produced 29% and 20% less sludge than EAO and COSA, respectively, simultaneously consumed 14% and 8% more oxygen than EAO and COSA, indicating that the sludge reduction in SOSA was not only caused by EAO-based aerobic digestion in the mainstream and conventional anaerobic reactions in the side-stream, but more importantly due to the bioaugmentation of sulfidogenesis. The roles of sulfidogenesis were further studied in batch tests, and the key findings were as follows: i) the SOSA biomass had a faster endogenous decay rate (0.097 d-1) than that of the COSA biomass (0.045 d-1), and ii) sulfidogenesis accelerated anaerobic solubilization, hydrolysis, acidogenesis and acetogenesis by 2.3 - 3.1 times, 6 - 22 %, 22 - 60% and 6 - 22%, respectively. Overall, the mechanisms of sludge reduction in SOSA were unraveled in this study which will help promote its full-scale application in future.

Coupling of sulfur(thiosulfate)-driven denitratation and anammox process to treat nitrate and ammonium contained wastewater.

This study investigated the feasibility of a new biological nitrogen removal process that integrates sulfur-driven autotrophic denitratation (NO3-→NO2-) and anaerobic ammonium oxidation (Anammox) for simultaneous removal of nitrate and ammonium from industrial wastewater. The proposed sulfur(thiosulfate)-driven denitratation and Anammox process was developed in two phases: First, the thiosulfate-driven denitratation was established in the UASB inoculated with activated sludge and fed with ammonium, nitrate and thiosulfate for 52 days until the nitrite level in the effluent reached 32.1 mg N/L. Second, enriched Anammox biomass was introduced to the UASB to develop the integrated thiosulfate-driven denitratation and Anammox (TDDA) bioprocess (53-212 d). Results showed that nitrate and ammonium could be efficiently removed from synthetic wastewater by the integrated TDDA system at a total nitrogen (TN) removal efficiency of 82.5 ±â€¯1.8% with an influent NH4+-N of 101.2 ±â€¯2.2 mgN/L, NO3--N of 101.1 ±â€¯1.5 mgN/L and thiosulfate of 202.5 ±â€¯3.2 mg S/L. It was estimated that Anammox and autotrophic denitritation (NO2-→N2) contributed to about 90% and 10% of the TN removal respectively at stable operation. The established TDDA system was further supported by high-throughput sequencing analysis that sulfur-oxidizing bacteria (e.g., Thiobacillus and Sulfurimonas) coexisted with Anammox bacteria (e.g., Ca. Kuenenia and Ca. Anammoxoglobus) in this syntrophic biocenosis. Additionally, batch experiments were conducted to reveal the kinetic rates and to reconcile the stoichiometry of the electron donor/acceptor couples of the TDDA process. The results unraveled the mechanisms in the new bioprocess: i) sulfite and elemental sulfur (S0) were initially generated from branched thiosulfate; ii) oxidation of sulfite and elemental sulfur coupled with fast and slow denitratation; iii) nitrite produced from denitratation together with ammonium were effectively converted to dinitrogen gas via Anammox.

Elucidating the biofilm properties and biokinetics of a sulfur-oxidizing moving-bed biofilm for mainstream nitrogen removal.

The sulfide-oxidizing autotrophic denitrification (SOAD) process offers a feasible alternative to mainstream heterotrophic denitrification in treating domestic sewage with insufficient organics. Previously SOAD has been successfully applied in a moving-bed biofilm reactor (MBBR). However, the biofilm properties and biokinetics are still not thoroughly understood. The present study was therefore designed to investigate these features of sulfur-oxidizing biofilms (SOBfs) cultivated in a lab-scale MBBR under stable operation for over a year. The biofilms developed were 160 µm thick, had an uneven and porous surface on which elemental sulfur (S0) accumulated, and the SOB biomass was highly diverse. The bioprocess kinetics were evaluated through 12 batch experiments. The results were interpreted by adopting a two-step sulfide oxidation model (sulfide→S0 and S0→ sulfate) with all specific rates having a linear regression coefficient of R2 > 0.9. Moreover, the inhibitory kinetic analysis revealed that 1) the maximum treatment capacity (about 480 mg S/(m2·h) and 80 mg N/(m2·h)) was observed at low sulfide level (40 mg S/L), while higher sulfide level (60-150 mg S/L) showed increasing inhibition on the oxidation of both sulfide and sulfur and denitrification. 2) The denitritation activity decreased by up to 43% when free nitrous acid reached a maximum of 8.6 µg N/L, whereas the oxidation of sulfide and sulfur did not have any significant effect. Interestingly, two physiologically diverse SOB groups were found in this special biofilm. The mechanisms of the cooperation and competition for electron donors and acceptors between these two SOB clades are proposed. The results of this study greatly enhance our understanding of the design and optimization of SOAD-MBBR for mainstream nitrogen removal.

Biological nitrogen removal from wastewater using sulphur-driven autotrophic denitrification.

Biological denitrification process in mainstream wastewater treatment often needs dosing supplemental electrons, consequently adding a remarkable operating cost. Organic carbon compounds are nowadays the most intensively used electron sources in full-scale wastewater treatment, corresponding with the well-understood carbon-nitrogen biogeochemistry for heterotrophic denitrification process. In the twenty-first century, the low-carbon technology is on calling to reduce the carbon footprint and relieve climate changing threatens. Autotrophic denitrification is highly recommended for mainstream wastewater treatment. The reduced-sulphur compounds (such as sulphide, elemental sulphur, and thiosulphate) could be utilised as electron donors, to drive sulphur cycle reactions to reduce nitrate and nitrite to dinitrogen gas. Based on the literature review and our own research experiences, this paper presents our perspectives on sulphur-driven autotrophic denitrification. It particularly focuses on the functional enzymes, sulphur bioreactors, and influential operating factors. Overall, this paper provides new insights on sulphur-nitrogen biogeochemistry and application as a low-carbon technology for nitrogen removal during municipal wastewater treatment.

Exploration of rapid start-up of the CANON process from activated sludge inoculum in a sequencing biofilm batch reactor (SBBR).

In this study, a laboratory-scale sequencing biofilm batch reactor (SBBR) was employed to explore a fast start-up of completely autotrophic nitrogen removal over nitrite (CANON) process. Partial nitrification was achieved by controlling free ammonia concentration and operating at above 30 °C; then the reactor was immediately operated with alternating periods of aerobiosis and anaerobiosis to start the anammox process. The CANON process was successfully achieved in less than 50 d, and the total-nitrogen removal efficiency and the nitrogen removal rate were 81% and 0.14 kg-N m(-3) d(-1) respectively. Afterwards, with the increasing of ammonium loading rate a maximum nitrogen removal rate of 0.39 kg-N m(-3) d(-1) was achieved on day 94. DNA analysis showed that 'Candidatus Brocadia' was the dominant anammox species and Nitrosomonas was the dominant aerobic ammonium-oxidizing bacteria in the CANON reactor. This study revealed that due to shortening the persistent and stable nitrite accumulation period the long start-up time of the CANON process can be significantly reduced.

Comparison of the MBBR denitrification carriers for advanced nitrogen removal of wastewater treatment plant effluent.

The moving bed biofilm reactors (MBBRs) were used to remove the residual NO3(-)-N of wastewater treatment plant (WWTP) effluent, and the MBBR carriers for denitrification were compared. The results showed that high denitrification efficiency can be achieved with polyethylene, polypropylene, polyurethane foam, and haydite carriers under following conditions: 7.2 to 8.0 pH, 24 to 26 °C temperature, 12 h hydraulic retention time (HRT), and 25.5 mg L(-1) external methanol dosage, while the WWTP effluent total nitrogen (TN) was between 2.6 and 15.4 mg L(-1) and NO3(-)-N was between 0.2 and 12.6 mg L(-1). The MBBR filled with polyethylene carriers had higher TN and NO3(-)-N removal rate (44.9 ± 19.1 and 83.4 ± 13.0%, respectively) than those with other carriers. The minimum effluent TN and NO3(-)-N of polyethylene MBBR were 1.6 and 0.1 mg L(-1), respectively, and the maximum denitrification rate reached 23.0 g m(-2) day(-1). When chemical oxygen demand (COD)/TN ratio dropped from 6 to 4, the NO3(-)- N and TN removal efficiency decreased significantly in all reactors except for that filled with polyethylene, which indicated that the polyethylene MBBR can resist influent fluctuation much better. The three-dimensional excitation-emission matrix analysis showed that all the influent and effluent of MBBRs contain soluble microbial products (SMPs)-like organics and biochemical oxygen demand (BOD), which can be removed better by MBBRs filled with haydite and polyethylene carriers. The nitrous oxide reductase (nosZ)-based terminal restriction fragment length polymorphism (T-RFLP) analysis suggested that the dominant bacteria in polyethylene MBBR are the key denitrificans.