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The coupling between anammox and nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) has been considered a sustainable technology for nitrogen removal from sidestream wastewater and can be implemented in both membrane biofilm reactor (MBfR) and granular bioreactor. However, the potential influence of the accompanying hydrogen sulfide (H 2 S) in the anaerobic digestion (AD)-related methane-containing mixture on anammox/n-DAMO remains unknown. To fill this gap, this work first constructed a model incorporating the C/N/S-related bioprocesses and evaluated/calibrated/validated the model using experimental data. The model was then used to explore the impact of H 2 S on the MBfR and granular bioreactor designed to perform anammox/n-DAMO at practical levels (i.e., 0∼5% (v/v) and 0∼40 g/S m 3 , respectively). The simulation results indicated that H 2 S in inflow gas did not significantly affect the total nitrogen (TN) removal of the MBfR under all operational conditions studied in this work, thus lifting the concern about applying AD-produced biogas to power up anammox/n-DAMO in the MBfR. However, the presence of H 2 S in the influent would either compromise the treatment performance of the granular bioreactor at a relatively high influent NH 4 + -N/NO 2 - -N ratio (e.g., >1.0) or lead to increased energy demand associated with TN removal at a relatively low influent NH 4 + -N/NO 2 - -N ratio (e.g., <0.7). Such a negative effect of the influent H 2 S could not be attenuated by regulating the hydraulic residence time and should therefore be avoided when applying the granular bioreactor to perform anammox/n-DAMO in practice., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Anaerobic digestion (AD) is a promising technique for waste management, which can achieve sludge stabilization and energy recovery. This study successfully prepared Fe 3 O 4 @ceramsite from WAS and applied it as an additive in sludge digestion, aiming to improve the conversion of organics to biomethane efficiency. Results showed that after adding the Fe 3 O 4 @ceramsite, the methane production was enhanced by 34.7% compared with the control group (88.0 ± 0.1 mL/g VS). Further mechanisms investigation revealed that Fe 3 O 4 @ceramsite enhanced digesta stability by strong buffering capacity, improved sludge conductivity, and promoted Fe (III) reduction. Moreover, Fe 3 O 4 @ceramsite has a larger surface area and better porous structure, which also facilitated AD performance. Microbial community analysis showed that some functional anaerobes related to AD such as Spirochaeta and Smithella were enriched with Fe 3 O 4 @ceramsite treatment. Potential syntrophic metabolisms between syntrophic bacteria (Syntrophomonas, associated with DIET) and methanogens were also detected in the Fe 3 O 4 @ceramsite treatment AD system., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Recently, the nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) processes have become a research hotspot in the field of wastewater treatment. The n-DAMO processes could not only mitigate direct and indirect carbon emissions from wastewater treatment plants but also strengthen biological nitrogen removal. However, the applications of n-DAMO-based biotechnologies face practical difficulties mainly caused by the distinctive properties of n-DAMO microorganisms and the limited/availability of methane with poor solubility. In this sense, the choice of bioreactors will play important roles that influence the growth and functioning of n-DAMO microorganisms, thus enabling dedicated development of the n-DAMO processes and efficient applications of n-DAMO-based biotechnologies. Therefore, this paper aims to discuss the three commonly-applied types of bioreactors, covering the individual working principle and state-of-the-art removal performance of nitrogen as well as dissolved methane observed when adopted for n-DAMO-based biotechnologies. With noted limitations for each bioreactor type, several key perspectives were proposed which hopefully would inspire future investigation and practical applications of the n-DAMO processes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Microbial denitrification is a crucial biological process for the treatment of nitrogen-polluted water. Traditional denitrification process consumes external organic carbon leading to an increase in treatment costs. We developed a novel sulfide-driven denitrification methane oxidation (SDMO) system that integrates autotrophic denitrification (AD) and denitrification anaerobic methane oxidation (DAMO) for cost-effective denitrification and biogas utilization in situ. Two SDMO systems were operated for 735 days, with nitrate and nitrite serving as electron acceptors, to explore the performance of sewage denitrification and characterize metabolic mechanisms. Results showed SDMO system could reach as high as 100% efficiency of nitrogen removal and biogas desulfurization without an external carbon source when HRT was 10 days and inflow nitrogen concentrations were 50-100 mgN·L -1 . Besides, nitrate was a preferable electron acceptor for SDMO system. Biogas not only enhanced nitrogen removal but also intensified the DAMO, nitrogen removed through DAMO contribution doubled as original period from 2.9 mgN·(L·d) -1 to 6.2 mgN·(L·d) -1 , and the ratio of nitrate removal through AD to DAMO was 1.2:1 with nitrate as electron acceptor. While nitrogen removed almost all through AD contribution and DAMO was weaken as before, the ratio of nitrate removal through AD to DAMO was 21.2:1 with nitrite as electron acceptor. Biogas introduced into SDMO system with nitrate inspired the growth of DAMO bacteria Candidatus Methylomirabilis from 0.3% to 19.6% and motivated its potentiality to remove nitrate without ANME archaea participation accompanying with gene mfnE upregulating ∼100 times. According to the reconstructed genome from binning analysis, the dramatically upregulated gene mfnE was derived from Candidatus Methylomirabilis, which may represent a novel metabolism pathway for DAMO bacteria to replace the role of archaea for nitrate reduction., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Integrating anammox with denitrifying anaerobic methane oxidation (DAMO) in the membrane biofilm reactor (MBfR) is a promising technology capable of achieving complete nitrogen removal from wastewater. However, it remains unknown whether reactor configurations featuring longitudinal gradients parallel to the membrane surface would affect the performance of the CH 4 -driven MBfR. To this end, this work aims to study the impacts of longitudinal heterogeneity potentially present in the gas and liquid phases on a representative CH 4 -driven MBfR performing anammox/DAMO by applying the reported modified compartmental modeling approach. Through comparing the modeling results of different reactor configurations, this work not only offered important guidance for better design, operation and monitoring of the CH 4 -driven MBfR, but also revealed important implications for prospective related modeling research. The total nitrogen removal efficiency of the MBfR at non-excessive CH 4 supply (e.g., surface loading of ≤0.064 g-COD m -2 d -1 in this work) was found to be insensitive to both longitudinal gradients in the liquid and gas phases. Comparatively, the longitudinal gradient in the liquid phase led to distinct longitudinal biomass stratification and therefore played an influential role in the effective CH 4 utilization efficiency, which was also related to the extent of reactor compartmentation considered in modeling. When supplied with non-excessive CH 4 , the MBfR is recommended to be designed/operated with both the biofilm reactor and the membrane lumen as plug flow reactors (PFRs) with co-current flow of wastewater and CH 4 , which could mitigate dissolved CH 4 discharge in the effluent. For the reactor configurations with the biofilm reactor designed/operated as a PFR, multi-spot sampling in the longitudinal direction is needed to obtain a correct representation of the microbial composition of the MBfR., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Denitrifying anaerobic methane oxidation (DAMO) coupled to anaerobic ammonium oxidation (anammox) is a promising technology for complete nitrogen removal with economic and environmental benefit. In this work, a model framework integrating DAMO and anammox process was constructed based on suspended-growth systems. The proposed model was calibrated and validated using experimental data from a sequencing batch reactor and a membrane aerated membrane bioreactor (MAMBR). The model managed to describe removal rates of ammonium (NH 4 + ), nitrite (NO 2 - ), and total nitrogen, as well as biomass changes of DAMO archaea, DAMO bacteria, and anaerobic ammonium oxidizing bacteria (AnAOB) in both reactors. The estimated parameter values revealed that DAMO archaea possessed properties of faster growth and higher biomass yield in suspended-growth systems compared to those in attached-growth systems (e.g., biofilm). Model simulation demonstrated that solid retention time (SRT) was effective in washing out DAMO bacteria, but retaining DAMO archaea and AnAOB in the MAMBR. The optimal SRT and nitritation efficiency (the ratio of the NO 2 - to the sum of NH 4 + and NO 2 - in the MAMBR influent) were simulated so that 99% of total nitrogen was removed to meet the discharge standard. MAMBR further suggested to be operated with SRT between 15 and 30 days so that the optimal nitritation efficiency could be minimized to 49% for cost savings.

Triclocarban (TCC), as the most typical antibacterial agent, is widely discovered in many ecological environment, especially in sludge. However, so far, no studies have reported the effect of TCC exposure on the properties of excess sludge. Therefore, in this study, TCC's toxicities to waste activated sludge (WAS) were analyzed by investigating the variation of physicochemical properties of sludge. It was found that TCC exposure has no effect on sludge pH, while it facilitated organic substances release from sludge, e.g. dissolved organic matter (DOM), protein and polysaccharide, which caused an increase of sludge reduction and changed the structure of functional groups and surface morphology of sludge. Moreover, we explored the effect of TCC on anaerobic digestion of WAS and found methane production was seriously inhibited by TCC. The related mechanism tests had illustrated that TCC exposure did not affect the hydrolysis process, but promoted the acidification and acetogenesis, and importantly inhibited the methanogenesis process. Methanogenic community was further evaluated and observed that the presence of TCC could vary the microbial community of methanogens with the abundance of aceticlastic methanogens increasing and hydrogenotrophic methanogens decreasing. These findings reached in this study would widen the understanding scope for TCC's toxicity to WAS., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Recently it has been shown that Candidatus 'Methanoperedens nitroreducens', an anaerobic methanotrophic archaea (ANME), can reduce nitrate to nitrite using electrons derived from anaerobic oxidation of methane. In this study, the growth kinetics of 'M. nitroreducens' enriched in a laboratory reactor were studied. In the experimental concentration range (up to 16 mg CH 4  L -1 ), anaerobic oxidation of methane by 'M. nitroreducens' was found to comply with first order kinetic model with a rate constant of 0.019 ± 0.006 h -1 and a biomass-specific rate constant of 0.04-0.14 L h -1  g -1 VSS. Meanwhile, the nitrate reduction to nitrite was well described by the Monod-type kinetic model with an affinity constant for nitrate of 2.1 ± 0.4 mg N L -1 , which is slightly higher than, but comparable to, that of most known denitrifying bacteria. This is the first time that the growth kinetics of 'M. nitroreducens' have been experimentally studied. The applicability of the kinetic model reported herein to this organism or similar organisms in natural or engineering systems requires further investigation., (Copyright © 2018. Published by Elsevier B.V.)

Anaerobic digestion of waste activated sludge (WAS) is often limited by low hydrolysis efficiencies and poor methane potentials. This work presents a novel pretreatment technology for WAS anaerobic digestion, i.e., combining free ammonia with heat pretreatment (CFHP). Experimental results showed that compared with control, solo free ammonia (135.4 mg NH 3 -N/L) and solo heat (70 °C) pretreatment, the combined free ammonia and heat (135.4 mg NH 3 -N/L with 70 °C) obtained 52.2%, 25.5% and 30.2% faster in hydrolysis rate and 25.2%, 17.9% and 16.5% higher in biochemical methane potential, respectively. Mechanism investigations showed that the combined pretreatment not only largely facilitated the disintegration of WAS but also increased the proportion of biodegradable organic matters, thereby providing better contract between biodegradable organics and the anaerobic microbes for methane production. Considering its effectiveness and renewability, the combined pretreatment is an attractive technology for the application in real-world situations., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Complete nitrogen removal has recently been demonstrated by integrating anaerobic ammonium oxidation (anammox) and denitrifying anaerobic methane oxidation (DAMO) processes. In this work, the effect of methane partial pressure on the performance of a membrane biofilm reactor (MBfR) consisting of DAMO and anammox microorganisms was evaluated. The activities of DAMO archaea and DAMO bacteria in the biofilm increased significantly with increased methane partial pressure, from 367 ± 9 and 58 ± 22 mg-N L -1 d -1 to 580 ± 12 and 222 ± 22 mg-N L -1 d -1 , respectively, while the activity of anammox bacteria only increased slightly, when the methane partial pressure was elevated from 0.24 to 1.39 atm in the short-term batch tests. The results were supported by a long-term (seven weeks) continuous test, when the methane partial pressure was dropped from 1.39 to 0.78 atm. The methane utilization efficiency was always above 96% during both short-term and long-term tests. Taken together, nitrogen removal rate (especially the nitrate reduction rate by DAMO archaea) and methane utilization efficiency could be maintained at high levels in a broad range of methane partial pressure (0.24-1.39 atm in this study). In addition, a previously established DAMO/anammox biofilm model was used to analyze the experimental data. The observed impacts of methane partial pressure on biofilm activity were well explained by the modeling results. These results suggest that methane partial pressure can potentially be used as a manipulated variable to control reaction rates, ultimately to maintain high nitrogen removal efficiency, according to nitrogen loading rate., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Reliable modelling of sulfide and methane production in sewer systems is required for efficient sewer emission management. Wastewater compositions affect sulfide and methane production kinetics through both its short-term variation influencing the substrate availability to sewer biofilms, and its long-term variation affecting the sewer biofilm structure. While the short-term effect is well considered in existing sewer models with the use of Monod or half-order equations, the long-term effect has not been explicitly considered in current sewer models suitable for network modelling. In this study, the long-term effect of wastewater compositions on sulfide and methane production activities in rising main sewers was investigated. A detailed biofilm model was firstly developed, and then calibrated and validated using experimental data measured during the entire biofilm development period of a laboratory sewer reactor. Based on scenario simulations using the detailed biofilm model, empirical equations describing the long-term effect of sulfate and sCOD (soluble chemical oxygen demand) concentrations on k H2S (the maximum sulfide production rate of sewer biofilm) and k CH4 (the maximum methane production rate of sewer biofilm) were proposed. These equations require further verification in future studies before their potential integration into network-wide sewer models., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Recent studies have shown that direct interspecies electron transfer (DIET) plays an important part in contributing to methane production from anaerobic digestion. However, so far anaerobic digestion models that have been proposed only consider two pathways for methane production, namely, acetoclastic methanogenesis and hydrogenotrophic methanogenesis, via indirect interspecies hydrogen transfer, which lacks an effective way for incorporating DIET into this paradigm. In this work, a new mathematical model is specifically developed to describe DIET process in anaerobic digestion through introducing extracellular electron transfer as a new pathway for methane production, taking anaerobic transformation of ethanol to methane as an example. The developed model was able to successfully predict experimental data on methane dynamics under different experimental conditions, supporting the validity of the developed model. Modeling predictions clearly demonstrated that DIET plays an important role in contributing to overall methane production (up to 33 %) and conductive material (i.e., carbon cloth) addition would significantly promote DIET through increasing ethanol conversion rate and methane production rate. The model developed in this work will potentially enhance our current understanding on syntrophic metabolism via DIET.

The discovery of denitrifying anaerobic methane oxidation (DAMO) processes enables the complete nitrogen removal from wastewater by utilizing the methane produced on site from anaerobic digesters. This model-based study investigated the mechanisms and operational window for efficient nitrogen removal by coupling nitritation-anaerobic ammonium oxidation (Anammox) and methane-dependent denitrification in membrane biofilm reactors (MBfRs). A mathematical model was applied to describe the microbial interactions among Anammox bacteria, DAMO archaea, and DAMO bacteria. The model sufficiently described the batch experimental data from an MBfR containing an Anammox-DAMO biofilm with different feeding nitrogen compositions, which confirmed the validity of the model. The effects of process parameters on the system performance and microbial community structure could therefore be reliably evaluated. The impacts of nitritation produced NO2(-)/NH4(+) ratio, methane supply, biofilm thickness and total nitrogen (TN) surface loading were comprehensively investigated with the model. Results showed that the optimum NO2(-)/NH4(+) ratio produced from nitritation for the Anammox-DAMO biofilm system was around 1.0 in order to achieve the maximum TN removal (over 99.0%), independent on TN surface loading. The corresponding optimal methane supply increased while the associated methane utilization efficiency decreased with the increase of TN surface loading. The cooperation between DAMO organisms and Anammox bacteria played the key role in the TN removal. Based on these results, the proof-of-concept feasibility of a single-stage MBfR coupling nitritation-Anammox-DAMO for complete nitrogen removal was also tested through integrating the model with ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) processes whilst controlling the dissolved oxygen (DO) concentration in the simulated system. The maximum TN removal was found to be achieved at the bulk DO concentration of around 0.17 g m(-3) under the simulation conditions, with the AOB, Anammox bacteria and DAMO organisms coexisting in the biofilm., (© 2015 Wiley Periodicals, Inc.)

In this study, the membrane biofilm reactor (MBfR) is proposed to achieve simultaneous removal of ammonium, dissolved methane, and sulfide from main-stream and side-stream anaerobic digestion liquors. To avoid dissolved methane stripping, oxygen is introduced through gas-permeable membranes, which also from the substratum for the growth of a biofilm likely comprising ammonium oxidizing bacteria (AOB), anaerobic ammonium oxidation (Anammox) bacteria, denitrifying anaerobic methane oxidation (DAMO) microorganisms, aerobic methane oxidizing bacteria (MOB), and sulfur oxidizing bacteria (SOB). A mathematical model is developed and applied to assess the feasibility of such a system and the associated microbial community structure under different operational conditions. The simulation studies demonstrate the feasibility of achieving high-level (>97.0%), simultaneous removal of ammonium, dissolved methane, and sulfide in the MBfRs from both main-stream and side-stream anaerobic digestion liquors through adjusting the influent surface loading (or hydraulic retention time (HRT)) and the oxygen surface loading. The optimal HRT was found to be inversely proportional to the corresponding oxygen surface loading. Under the optimal operational conditions, AOB, DAMO bacteria, MOB, and SOB dominate the biofilm of the main-stream MBfR, while AOB, Anammox bacteria, DAMO bacteria, and SOB coexist in the side-stream MBfR to remove ammonium, dissolved methane, and sulfide simultaneously.

Sewer sediment processes have been reported to significantly contribute to overall sulfide and methane production in sewers, at a scale comparable to that of sewer biofilms. The physiochemical and biological characteristics of sewer sediments are heterogeneous; however, the variability of in-sediments sulfide and methane production rates among sewers has not been assessed to date. In this study, five sewer sediment samples were collected from two cities in Australia with different climatic conditions. Batch assays were conducted to determine the rates of sulfate reduction and methane production under different flow velocity (shear stress) conditions as well as under completely mixed conditions. The tests showed substantial and variable sulfate reduction and methane production activities among different sediments. Sulfate reduction and methane production from sewer sediments were confirmed to be areal processes, and were dependent on flow velocity/shear stress. Despite of the varying characteristics and reactions kinetics, the sulfate reduction and methane production processes in all sediments could be well described by a one-dimensional sewer sediment model recently developed based on results obtained from a laboratory sewer sediment reactor. Model simulations indicated that the in-situ contribution of sewer sediment emissions could be estimated without the requirement of measuring the specific sediment characteristics or the sediment depths., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The presence of a high level of dissolved methane (e.g., 20-26 g m(-3)) in the anaerobic sludge digestion liquor represents a major challenge to the treatment of this stream, as its emission to the atmosphere contributes significantly to the carbon footprint of wastewater treatment. Here we propose a new approach to simultaneous ammonium and dissolved methane removal from the anaerobic digestion liquor through integrating partial nitritation-Anammox and denitrifying anaerobic methane oxidation (DAMO) processes in a single-stage membrane biofilm reactor (MBfR). In such an MBfR, the anaerobic digestion liquor is provided in the bulk liquid, while oxygen is supplied through gas-permeable membranes to avoid dissolved methane stripping. A previously developed model with appropriate extensions was applied to assess the system performance under different operational conditions and the corresponding microbial interactions. Both influent surface loading (or hydraulic retention time) and oxygen surface loading are found to significantly influence the total nitrogen (TN) and dissolved methane removal, which jointly determine the overall system performance. The counter diffusion and concentration gradients of substrates cause microbial stratification in the biofilm, where ammonia-oxidizing bacteria (AOB) attach close to the membrane surface (biofilm base) where oxygen and ammonium are available, while Anammox and DAMO microorganisms jointly grow in the biofilm layer close to the bulk liquid where methane, ammonium, and nitrite are available with the latter produced by AOB. These results provide first insights and useful information for the design and operation of this new technology for simultaneous ammonium and dissolved methane removal in its potential future applications., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Recent studies have shown that sewer systems produce and emit a significant amount of methane. Methanogens produce methane under anaerobic conditions in sewer biofilms and sediments, and the stratification of methanogens and sulfate-reducing bacteria may explain the simultaneous production of methane and sulfide in sewers. No significant methane sinks or methanotrophic activities have been identified in sewers to date. Therefore, most of the methane would be emitted at the interface between sewage and atmosphere in gravity sewers, pumping stations, and inlets of wastewater treatment plants, although oxidation of methane in the aeration basin of a wastewater treatment plant has been reported recently. Online measurements have also revealed highly dynamic temporal and spatial variations in methane production caused by factors such as hydraulic retention time, area-to-volume ratio, temperature, and concentration of organic matter in sewage. Both mechanistic and empirical models have been proposed to predict methane production in sewers. Due to the sensitivity of methanogens to environmental conditions, most of the chemicals effective in controlling sulfide in sewers also suppress or diminish methane production. In this paper, we review the recent studies on methane emission from sewers, including the production mechanisms, quantification, modeling, and mitigation., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Zero valent iron (ZVI) packed anaerobic granular sludge reactors have been developed for improved anaerobic wastewater treatment. In this work, a mathematical model is developed to describe the enhanced methane production and sulfate reduction in anaerobic granular sludge reactors with the addition of ZVI. The model is successfully calibrated and validated using long-term experimental data sets from two independent ZVI-enhanced anaerobic granular sludge reactors with different operational conditions. The model satisfactorily describes the chemical oxygen demand (COD) removal, sulfate reduction and methane production data from both systems. Results show ZVI directly promotes propionate degradation and methanogenesis to enhance methane production. Simultaneously, ZVI alleviates the inhibition of un-dissociated H2S on acetogens, methanogens and sulfate reducing bacteria (SRB) through buffering pH (Fe(0) + 2H(+) = Fe(2+) + H2) and iron sulfide precipitation, which improve the sulfate reduction capacity, especially under deterioration conditions. In addition, the enhancement of ZVI on methane production and sulfate reduction occurs mainly at relatively low COD/ [Formula: see text] ratio (e.g., 2-4.5) rather than high COD/ [Formula: see text] ratio (e.g., 16.7) compared to the reactor without ZVI addition. The model proposed in this work is expected to provide support for further development of a more efficient ZVI-based anaerobic granular system., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Nitrate dosing is widely used to control sulfide and methane formation in sewers. The impact of nitrate on sulfide and methane production by sewer biofilms in rising mains has been elucidated recently. However, little is known about the effect of nitrate on biologically active sewer sediment, which is substantially thicker than rising main biofilms (centimeters vs. hundreds of micrometers, respectively). In this study, we investigated the effect of nitrate addition to sewer sediment cultivated in lab-scale sewer sediment reactors. Batch test results showed that nitrate addition does not suppress sulfide production in sewer sediment, but it reduces sulfide accumulation through anoxic sulfide oxidation in the sediment and hence, also reduces sulfide accumulation in the bulk water. Microsensor measurement of sediment sulfide revealed the presence of sulfide oxidation and sulfide production zones with the interface dynamically regulated by the depth of nitrate penetration. In contrast, the methane production activity of sewer sediment was substantially reduced, likely due to the long-term inhibitory effects of nitrate on methanogens. Pore water measurements showed that methane production activity in the sediment zone with frequent nitrate exposure was completely suppressed, and consequently, the methane production zone re-established deeper in the sediment where nitrate penetration was infrequent., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Background: Anaerobic digestion has been widely applied to treat the waste activated sludge from biological wastewater treatment and produce methane for biofuel, which has been one of the most efficient solutions to both energy crisis and environmental pollution challenges. Anaerobic digestion sludge contains highly complex microbial communities, which play crucial roles in sludge treatment. However, traditional approaches based on 16S rRNA amplification or fluorescent in situ hybridization cannot completely reveal the whole microbial community structure due to the extremely high complexity of the involved communities. In this sense, the next-generation high-throughput sequencing provides a powerful tool for dissecting microbial community structure and methane-producing pathways in anaerobic digestion., Results: In this work, the metagenomic sequencing was used to characterize microbial community structure of the anaerobic digestion sludge from a full-scale municipal wastewater treatment plant. Over 3.0 gigabases of metagenomic sequence data were generated with the Illumina HiSeq 2000 platform. Taxonomic analysis by MG-RAST server indicated that overall bacteria were dominant (~93%) whereas a considerable abundance of archaea (~6%) were also detected in the anaerobic digestion sludge. The most abundant bacterial populations were found to be Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria. Key microorganisms and related pathways involved in methanogenesis were further revealed. The dominant proliferation of Methanosaeta and Methanosarcina, together with the functional affiliation of enzymes-encoding genes (acetate kinase (AckA), phosphate acetyltransferase (PTA), and acetyl-CoA synthetase (ACSS)), suggested that the acetoclastic methanogenesis is the dominant methanogenesis pathway in the full-scale anaerobic digester., Conclusions: In short, the metagenomic sequencing study of this work successfully dissected the detail microbial community structure and the dominated methane-producing pathways of a full-scale anaerobic digester. The knowledge garnered would facilitate to develop more efficient full-scale anaerobic digestion systems to achieve high-rate waste sludge treatment and methane production.

Recent studies have demonstrated significant sulfide and methane production by sewer biofilms, particularly in rising mains. Sewer sediments in gravity sewers are also biologically active; however, their contribution to biological transformations in sewers is poorly understood at present. In this study, sediments collected from a gravity sewer were cultivated in a laboratory reactor fed with real wastewater for more than one year to obtain intact sediments. Batch test results show significant sulfide production with an average rate of 9.20 ± 0.39 g S/m(2)·d from the sediments, which is significantly higher than the areal rate of sewer biofilms. In contrast, the average methane production rate is 1.56 ± 0.14 g CH4/m(2)·d at 20 °C, which is comparable to the areal rate of sewer biofilms. These results clearly show that the contributions of sewer sediments to sulfide and methane production cannot be ignored when evaluating sewer emissions. Microsensor and pore water measurements of sulfide, sulfate and methane in the sediments, microbial profiling along the depth of the sediments and mathematical modelling reveal that sulfide production takes place near the sediment surface due to the limited penetration of sulfate. In comparison, methane production occurs in a much deeper zone below the surface likely due to the better penetration of soluble organic carbon. Modelling results illustrate the dependency of sulfide and methane productions on the bulk sulfate and soluble organic carbon concentrations can be well described with half-order kinetics., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Anaerobic digestion has been widely applied for waste activated sludge (WAS) treatment. However, methane production from anaerobic digestion of WAS is usually limited by the slow hydrolysis rate and/or poor biochemical methane potential of WAS. This work systematically studied the effects of three different types of zero valent iron (i.e., iron powder, clean scrap and rusty scrap) on methane production from WAS in anaerobic digestion, by using both experimental and mathematical approaches. The results demonstrated that both the clean and the rusty iron scrap were more effective than the iron powder for improving methane production from WAS. Model-based analysis showed that ZVI addition significantly enhanced methane production from WAS through improving the biochemical methane potential of WAS rather than its hydrolysis rate. Economic analysis indicated that the ZVI-based technology for enhancing methane production from WAS is economically attractive, particularly considering that iron scrap can be freely acquired from industrial waste. Based on these results, the ZVI-based anaerobic digestion process of this work could be easily integrated with the conventional chemical phosphorus removal process in wastewater treatment plant to form a cost-effective and environment-friendly approach, enabling maximum resource recovery/reuse while achieving enhanced methane production in wastewater treatment system.

Methanethiol (MT) is considered one of the predominant odorants in sewer systems. Therefore, understanding MT transformation in sewers is essential to sewer odor assessment and abatement. In this study, we investigated the degradation of MT in laboratory anaerobic sewers. Experiments were carried out in seven anaerobic sewer reactors with biofilms at different stages of development. MT degradation was found to be strongly dependent on the methanogenic activity of sewer biofilms. The MT degradation rate accelerated with the increase of methanogenic activity of sewer biofilms, resulting in MT accumulation (i.e. net production) in sewer reactors with relatively low methanogenic activities, and MT removal in reactors with higher methanogenic activities. A Monod-type kinetic expression was developed to describe MT degradation kinetics in anaerobic sewers, in which the maximum degradation rate was modeled as a function of the maximum methane production rate through a power function. It was also found that MT concentration had a linear relationship with acetate concentration, which may be used for preliminary assessment of MT presence in anaerobic sewers., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Simultaneous production of sulfide and methane by anaerobic sewer biofilms has recently been observed, suggesting that sulfate-reducing bacteria (SRB) and methanogenic archaea (MA), microorganisms known to compete for the same substrates, can coexist in this environment. This study investigated the community structures and activities of SRB and MA in anaerobic sewer biofilms (average thickness of 800 μm) using a combination of microelectrode measurements, molecular techniques, and mathematical modeling. It was seen that sulfide was mainly produced in the outer layer of the biofilm, between the depths of 0 and 300 μm, which is in good agreement with the distribution of SRB population as revealed by cryosection-fluorescence in situ hybridization (FISH). SRB had a higher relative abundance of 20% on the surface layer, which decreased gradually to below 3% at a depth of 400 μm. In contrast, MA mainly inhabited the inner layer of the biofilm. Their relative abundances increased from 10% to 75% at depths of 200 μm and 700 μm, respectively, from the biofilm surface layer. High-throughput pyrosequencing of 16S rRNA amplicons showed that SRB in the biofilm were mainly affiliated with five genera, Desulfobulbus, Desulfomicrobium, Desulfovibrio, Desulfatiferula, and Desulforegula, while about 90% of the MA population belonged to the genus Methanosaeta. The spatial organizations of SRB and MA revealed by pyrosequencing were consistent with the FISH results. A biofilm model was constructed to simulate the SRB and MA distributions in the anaerobic sewer biofilm. The good fit between model predictions and the experimental data indicate that the coexistence and spatial structure of SRB and MA in the biofilm resulted from the microbial types and their metabolic transformations and interactions with substrates., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

Nitrogen removal by using the synergy of denitrifying anaerobic methane oxidation (DAMO) and anaerobic ammonium oxidation (Anammox) microorganisms in a membrane biofilm reactor (MBfR) has previously been demonstrated experimentally. In this work, a mathematical model is developed to describe the simultaneous anaerobic methane and ammonium oxidation by DAMO and Anammox microorganisms in an MBfR for the first time. In this model, DAMO archaea convert nitrate, both externally fed and/or produced by Anammox, to nitrite, with methane as the electron donor. Anammox and DAMO bacteria jointly remove the nitrite fed/produced, with ammonium and methane as the electron donor, respectively. The model is successfully calibrated and validated using the long-term (over 400 days) dynamic experimental data from the MBfR, as well as two independent batch tests at different operational stages of the MBfR. The model satisfactorily describes the methane oxidation and nitrogen conversion data from the system. Modeling results show the concentration gradients of methane and nitrogen would cause stratification of the biofilm, where Anammox bacteria mainly grow in the biofilm layer close to the bulk liquid and DAMO organisms attach close to the membrane surface. The low surface methane loadings result in a low fraction of DAMO microorganisms, but the high surface methane loadings would lead to overgrowth of DAMO bacteria, which would compete with Anammox for nitrite and decrease the fraction of Anammox bacteria. The results suggest an optimal methane supply under the given condition should be applied not only to benefit the nitrogen removal but also to avoid potential methane emissions.

Nitrate (NO 3 − ) and arsenite (As(III)) are two major contaminants in groundwater, which could cause significant risks to human wellbeing and ecological system. In this work, a single-stage membrane biofilm reactor (MBfR) coupling denitrifying anaerobic methane (CH 4 ) oxidation (DAMO) and autotrophic As(III) oxidation processes was proposed for the first time to achieve the in-situ or ex-situ simultaneous removal of NO 3 − and As(III) from groundwater. CH 4 is supplied to the MBfR through gas-permeable membranes while NO 3 − and As(III) are provided in the bulk liquid. A mathematical model was developed by integrating the well-established biokinetics of DAMO microorganisms with the kinetics of As(III)-oxidizing bacteria (AsOB). The key parameter values of AsOB were specifically estimated using the batch experimental data of an enriched pure AsOB culture in conjunction with thermodynamic state calculations. The maximum specific growth rate of AsOB ( μ AsOB ) and the yield coefficient for AsOB ( Y AsOB ) were determined to be 0.00161 h −1 and 0.016 g COD g −1 As, respectively. The modeling results demonstrated that both influent surface loading (or hydraulic retention time (HRT)) and CH 4 surface loading played important roles in controlling the steady-state microbial community structure and thus significantly affected the system performance. The As(III)/NO 3 − ratio between 0.1 and 2 g As g −1 NO 3 − -N in the influent would have no significant impact on the overall system performance despite the varying microbial composition in the biofilm. Through properly adjusting the influent surface loading (or HRT) and CH 4 surface loading whilst maintaining a sufficient biofilm thickness at a suitable influent As(III)/NO 3 − ratio, the maximum removal efficiencies of total nitrogen and As(III) could both reach above 95.0%, accompanied by a high CH 4 utilization efficiency of up to 99.0%. [ABSTRACT FROM AUTHOR]

• BMPs aggravate N 2 O and CH 4 emissions from sediments more severely than NBMPs. • BMPs facilitate denitrification, while NBMPs inhibit nitrogen metabolism. • Nitrifier denitrification dominates over NH 2 OH oxidation after BMPs exposure. • Microplastics promote hydrogen-dependent methanogenic pathway. Freshwater ecosystems, such as urban lake sediments, have been identified as important sources of greenhouse gases (GHGs) to the atmosphere, as well as persistent sinks for ubiquitous microplastics due to the high population density and frequent anthropogenic activity. The potential impacts of microplastics on GHG production, however, remain underexplored. In this study, four types of common biodegradable microplastics (BMPs) versus four conventional non-biodegradable microplastics (NBMPs) were artificially exposed to urban lake sediments to investigate the responses of nitrous oxide (N 2 O) and methane (CH 4) production, and make a comparison regarding how the biodegradability of microplastics affected GHG emissions. Importantly, results suggested that BMPs aggravated N 2 O and CH 4 production in urban lake sediments more severely than conventional NBMPs. The production rates of N 2 O and CH 4 increased by 48.78–71.88 % and 30.87–69.12 %, respectively, in BMPs groups, while those increased by only 0–25.69 % and 6.46–10.46 % with NBMPs exposure. Moreover, BMPs insignificantly affected nitrification but facilitated denitrification, while NBMPs inhibited both processes. BMPs not only created more oxygen-limited microenvironment, greatly promoting N 2 O production via nitrifier denitrification pathway, but also provided dissolved organic carbon favoring heterotrophic denitrification, which was primarily supported by the enriched denitrifiers and functional genes. In contrast, NBMPs slightly upregulated nitrifier denitrification pathway to generate N 2 O, and showed a toxic inhibition on both nitrifiers and denitrifiers. In addition, both BMPs and NBMPs promoted hydrogen-dependent methanogenic pathway but suppressed acetate-dependent pathway. The greater enhancement of CH 4 production with BMPs exposure was attributed to the additional organic carbon substrates derived from BMPs and the stimulated microbial methane metabolism activities. [Display omitted] [ABSTRACT FROM AUTHOR]

Methane recovery from waste activated sludge (WAS) through anaerobic digestion is generally restricted by the poor degradability of WAS. Herein, a novel sludge pretreatment technology by using the calcium hypochlorite (Ca(ClO) 2) in enhancing the methane production from WAS anaerobic digestion was reported. The solubilization of WAS was significantly increased after 10–240 mg Ca(ClO) 2 /g VS (VS: volatile solids) pretreatment for 48 h, under which the solubilization was 1.7–3.4 folds (i.e., 0.17–0.34 mg SCOD/mg VS; SCOD: soluble chemical oxygen demand) higher than that without Ca(ClO) 2 pretreatment (i.e., 0.1 mg SCOD/mg VS). Correspondingly, the methane production was increased from 250.0 ± 5.3 mL/g VS to 385.1 ± 3.3 mL/g VS with the doses of Ca(ClO) 2 increasing from 10 mg/g VS to 240 mg/g VS, resulted in an increasing methane production of 3.6%–59.7% than that without Ca(ClO) 2 pretreatment. The microbial community composition results exhibited that the populations of key acidogens (e.g., Longilinea sp.) and methanogens (e.g., Methanosaeta sp.) were both reduced significantly. Moreover, Ca(ClO) 2 decreased the cells viability, leading to a 76.2% reduction of living cells fraction. Accordingly, it was further confirmed that high dosage of Ca(ClO) 2 could inhibit three microbial-related processes relevant to methane production, i.e., acidification, hydrolysis and methanogenesis. [Display omitted] • Ca(ClO) 2 pretreatment enhanced methane production from WAS digestion. • Methane yield is positively related to pretreatment level of Ca(ClO) 2. • High dosage of Ca(ClO) 2 decreased the abundance of acidogens and methanogens. • High dosage of Ca(ClO) 2 inhibited hydrolysis, acidification, and methanogenesis. [ABSTRACT FROM AUTHOR]

Previous studies reported that free ammonia (FA) pretreatment could improve methane production from anaerobic digestion of waste activated sludge (WAS) effectively. However, details of how FA pretreatment improves methane production are poorly understood. This study therefore aims to reveal the underlying mechanisms of FA pretreatment affecting anaerobic digestion of WAS through a series of batch tests using either real sludge or synthetic media as the digestion substrates at different pH values. At pH 8.5 level, with an increase of FA level from 18.5 to 92.5 mg/L (i.e., NH+ 4-N: 100–500 mg/L; pH 8.5) the maximum methane yield varied between 194.0 ± 3.9 and 196.9 ± 7.7 mL/g of VSS (25 °C, 1 atm). At pH 9.5 or 10 level, however, with an increase of initial FA level from 103.2 to 516.2 mg/L, the maximal methane yield increased linearly. The mechanism studies revealed that FA pretreatment at high levels not only accelerated the disintegration of WAS but also enhanced the biodegradability of WAS. Although pH in the digesters was adjusted to 7.0 ± 0.1, the high levels of NH+ 4-N added or released led to substantial levels of residual FA ranging from 4.4 to 11.6 mg/L. It was found that this level of FA inhibited homoacetogenesis and methanogenesis significantly, though hydrolysis, acidogenesis, and acetogenesis processes were unaffected largely. Further analyses showed that the inhibition constant of FA to substrate degradation was in the sequence of dextran > glucose > hydrogen > acetate, indicating the methanogenesis process was more sensitive to FA. [ABSTRACT FROM AUTHOR]

[Display omitted] • Ideal influent substrates conditions for n-DAMO/anammox-based granular bioreactor. • Smaller granules rich in n-DAMO archaea/AnAOB accelerated bioreactor start-up. • Low diffusivity of solutes promoted bioreactor start-up and CH 4 utilization. In this work, a bioprocess model was applied to first determine the impacts of influent substrates conditions on the granular bioreactor performing nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) and anammox integrated processes and then investigate the roles of granular sludge properties in regulating the bioreactor performance and start-up process. The ideal influent substrates conditions were identified at NO 2 −-N/NH 4 +-N of 1:1 and dissolved CH 4 concentration of 85 g COD m−3, which achieved 98.6% total nitrogen removal and 87.7% dissolved CH 4 utilization. Under such ideal influent conditions, the initial properties of granular sludge didn't significantly affect the granular bioreactor performance. However, inoculation of granular sludge with a relatively small granular sludge size and a high abundance of n-DAMO archaea or/and anammox bacteria could effectively shorten the bioreactor start-up. Meanwhile, reducing the diffusivity of solutes within granular sludge was also beneficial for expediting the start-up process and promoting dissolved CH 4 utilization. [ABSTRACT FROM AUTHOR]

Perchlorate (ClO 4 − ) is recognized as an important contaminant in surface water and groundwater, which would pose health risks at very low concentrations. A methane-based membrane biofilm reactor (MBfR) has been successfully demonstrated for perchlorate reduction, which provided an alternative solution for perchlorate remediation with low cost. In this work, a multispecies biofilm model was developed to evaluate perchlorate reduction in the methane-based MBfR under different operational conditions. The model was calibrated and validated using the experimental data from the long-term operation of the MBfR at seven distinct stages. The results suggested that the developed model could satisfactorily describe perchlorate reduction and denitrification performances in the MBfR (R 2 > 0.9). The modeling results provided insight into the microbial community distribution in the biofilm, with aerobic methanotrophs and perchlorate reduction bacteria being mainly located at the membrane side (∼60%) and heterotrophic bacteria being situated near the liquid side (∼50%). The model simulations indicated that over 80% of perchlorate removal efficiency could be achieved through controlling the optimal combinations of methane pressure (P CH4 ) and perchlorate loading (L ClO4 ) (e.g., applying a P CH4 of 30 kPa at a L ClO4 of 0.08 g Cl/m 2 /d). In addition, the perchlorate reduction would be inhibited by the presence of nitrate and nitrite in the MBfR, which should be appropriately controlled during the future practical application of the promising process. [ABSTRACT FROM AUTHOR]

Recent studies have investigated the potential of enhanced groundwater Vinyl Chloride (VC) remediation in the presence of methane and ethene through the interactions of VC-assimilating bacteria, methanotrophs and ethenotrophs. In this study, a mathematical model was developed to describe aerobic biotransformation of VC in the presence of methane and ethene for the first time. It examines the metabolism of VC by VC-assimilating bacteria as well as cometabolism of VC by both methanotrophs and ethenotrophs, using methane and ethene respectively, under aerobic conditions. The developed model was successfully calibrated and validated using experimental data from microcosms with different experimental conditions. The model satisfactorily describes VC, methane and ethene dynamics in all microcosms tested. Modeling results describe that methanotrophic cometabolism of ethene promotes ethenotrophic VC cometabolism, which significantly enhances aerobic VC degradation in the presence of methane and ethene. This model is expected to be a useful tool to support effective and efficient processes for groundwater VC remediation. [ABSTRACT FROM AUTHOR]

Hydrogen sulfide (H 2 S) produced by biofilms and sediments in sewer systems causes severe odor and corrosion problems, resulting in billions of dollars loss annually in the world. Meanwhile, methane (CH 4) production by anaerobic process in sewers may incur sewer explosion and greenhouse gas emission. Among all current sewer management measures, iron compounds have been widely used due to their effective impacts, non-toxicity, and relatively low cost. Owing to unique bio-chemical properties, the Fe (Ⅱ)-Fe (Ⅲ) iron cycle is also involved with the conversion of C, N, P and some other bioprocesses in sewer systems. In this review, the possible redox reactions in sewer systems related to bio-chemical iron cycle are comprehensively summarized, including sulfate oxidation, organic carbon oxidation, anaerobic ammonium oxidation and denitrification. Afterwards, the effects of iron compounds addition in sewer systems on nitrogen, phosphorus and organic pollutants removal are reviewed. Critical perspectives regarding future directions of iron assisted sewer management have been proposed to guide further research. This review is expected to provide instructive insights into the function of bio-chemical iron cycle in sewer systems. [Display omitted] • The mechanism of bio-chemical iron cycle in sewer systems is clarified. • The iron cycle inhibits H 2 S and CH 4 production and emissions. • Iron addition in sewers enhances the removal of C, N, and P. • Ferric ions improve the key sludge properties in sewer systems. [ABSTRACT FROM AUTHOR]

• Prepared novel conductive polypyrrole to improve the anaerobic sludge digestion performance. • The accumulative methane production was increased by 27.83% by adding Ppy. • Metatranscriptomic analysis illustrated Ppy promoted acidogenesis and methanogenesis. • Ppy enhances the utilization of acids and electrons by methanogens through DIET. Anaerobic digestion (AD) is a promising method for treating waste activated sludge (WAS), but the low methane yield limits its large-scale application. The addition of conductive nanomaterials has been demonstrated to enhance the activity of AD via promoting the direct interspecies electron transfer (DIET). In this study, novel conductive polypyrrole (Ppy) was prepared to effectively improve the AD performance of WAS. The results showed that the accumulative methane production was enhanced by 27.83% by Ppy, with both acidogenesis and methanogenesis being efficiently accelerated. The microbial community analysis indicated that the abundance of bacteria associated with acidogenesis process was significantly elevated by Ppy. Further investigation by metatranscriptomics revealed that fadE and fadN genes (to express the key enzymes in fatty acid metabolism) were highly expressed in the Ppy-driven AD, suggesting that Ppy promoted electron generation during acid production. For methanogenesis metabolism, genes related to acetate utilization and CO 2 utilization methanogenesis were also up-regulated by Ppy, illustrating that Ppy facilitates the utilization of acetate and electrons by methanogenic archaea, thus potentially promoting the methanogenesis through DIET. [Display omitted] [ABSTRACT FROM AUTHOR]

• Methane production was predicted by AutoML algorithms based on microplastic dataset. • GBM-based models showed better prediction performance than neural networks. • The order of variable importance was determined by integrated explainable methods. • The variable of microplastic types dominated the prediction of methane production. • Increasing microplastic diameter and concentration inhibited methane production. Microplastics as emerging pollutants have been heavily accumulated in the waste activated sludge (WAS) during biological wastewater treatment, which showed significantly diverse impacts on the subsequent anaerobic sludge digestion for methane production. However, a robust modeling approach for predicting and unveiling the complex effects of accumulated microplastics within WAS on methane production is still missing. In this study, four automated machine learning (AutoML) approach was applied to model the effects of microplastics on anaerobic digestion processes, and integrated explainable analysis was explored to reveal the relationships between key variables (e.g. , concentration, type, and size of microplastics) and methane production. The results showed that the gradient boosting machine had better prediction performance (mean squared error (MSE) = 17.0) than common neural networks models (MSE = 58.0), demonstrating that the AutoML algorithms succeeded in predicting the methane production and could select the best machine learning model without human intervention. Explainable analysis results indicated that the variable of microplastic types was more important than the variable of microplastic diameter and concentration. The existence of polystyrene was associated with higher methane production, whereas increasing microplastic diameter and concentration both inhibited methane production. This work also provided a novel modeling approach for comprehensively understanding the complex effects of microplastics on methane production, which revealed the dependence relationships between methane production and key variables and may be served as a reference for optimizing operational adjustments in anaerobic digestion processes. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] • Alkaline pre-fermentation (AF) enhanced polyacrylamide (PAM) degradation to 80.1%. • AF accelerated the hydrolysis and acidification of PAM-flocculated sludge (PFS). • Biochemical methane potential of PFS was improved by ~230% through AF. • AF broke the large firm "PAM-sludge" floccules and decreased PAM molecular weight. • Petrimonas and Sedimentibacter and Methanosaeta were enriched in AF-digester. The residual Polyacrylamide (PAM) in sewage sludge might cause severe disturbance in anaerobic digestion, and appropriate solutions to alleviate such situation are urgently required. In present study, alkaline pre-fermentation was proposed for PAM-flocculated sewage sludge (PFS) pretreatment, by which both PFS methane production and PAM degradation were remarkably enhanced. Under the optimal alkaline pre-fermentation condition (pH 10 for 12 d), the biochemical methane potential of PFS (12 g PAM/kg TS) increased from 107.2 to 246.6 mL/g VS, the hydrolysis rate increased from 0.109 to 0.197 d−1, and the degradation efficiency of PAM increased from 30.6% to 80.1%. Mechanism analysis indicated that the alkaline pre-fermentation broke the large "PAM-sludge" floccules, decreased the molecular weight of PAM, which alleviated the disturbance situation of PAM-present digester and made PAM more available for microbes to be biodegraded. Moreover, PFS hydrolysis and acidification were simultaneously accelerated by alkaline pre-fermentation, thereby providing more bioavailable carbon substrates for subsequent methane producing and PAM co-metabolism. Microbial community analysis demonstrated syntrophic bacteria such as Petrimonas and Sedimentibacter , which had ability to degrade an extensive range of various types of organics including carbohydrates and PAM, were enriched in alkaline pre-fermenter, and the acetotrophic methanogen Methanosaeta , were elevated in anaerobic digester. This work provides an effective microbial based strategy to improve the efficiency of anaerobic digestion of PFS. [ABSTRACT FROM AUTHOR]

An innovative technology using diatomite to enhance methane production from anaerobic algae digestion was proposed in this study. The results of biochemical methane production (BMP) tests showed that 1, 2, 5 g/L of diatomite addition increased methane production by 22.5 ± 7.1%, 30.6 ± 5.1% and 32.8 ± 7.2% (p = 0.001, 0.0002, and 0.0002) compared to the control. In continuous operation experiments over 136 days, the parameters of digester containing volatile solid (VS) destruction and daily methane yield were increased by 31.2% and 32.1% with diatomite (5 g/L) addition. The addition of diatomite improved the solubilization, hydrolysis, acidification, and methanation processes in the anaerobic algae digestion, and the abundances of most key microorganisms (e.g., Methanosaeta sp. and Syntrophomonas sp.) involved in anaerobic digestion also increased correspondingly. The mechanism studies revealed that diatomite with strong conductivity could act as an electron shuttle, contributing to the increasing electron transfer efficiency for methane production. The novel strategy proposed in this study might bring significant economic and environmental benefits to algae treatment for energy recovery. [Display omitted] • Diatomite enhanced methane production from anaerobic algae digestion. • Diatomite promoted VS destruction and daily methane yield in continuous test. • Diatomite improved the solubilization, hydrolysis, acidification and methanation. • Diatomite enhanced the richness of key microbial community. • Diatomite increased electron transfer efficiency for methane production. [ABSTRACT FROM AUTHOR]

• The reflux ratio of 20% digestion liquid increased the methane production from WAS by 1.6 times. • The digestion liquid promoted the decomposition of WAS and improved the biochemical methane potential. • Ammonium and metal ions increase methane production by promoting WAS solubilization or activating enzyme expression. • The refractory organics reduced methane yield through suppressing the bio-processes. This work proved an efficient method to significantly increase methane production from anaerobic digestion of WAS. This method is to reflux proper of digestion liquid into waste activated sludge pretreatment unit (pH 9.5 for 24 h). The yield of maximum methane improved between 174.2 ± 7.3 and 282.5 ± 14.1 mL/g VSS with the reflux ratio of digestion liquid increasing from 0% to 20%. It was observed that the biodegradable organics in the digestion liquid did not affect the biological processes related to anaerobic digestion but increased methane production through reutilization. The ammonium in the digestion liquid was the main contributor to the increase in methane production via promoting sludge solubilization, but refractory organics were the major inhibitors to anaerobic digestion. It should be emphasized that the metal ions present in the digestion liquid were beneficial rather than harmful to the biological processes in the anaerobic digestion, which may be connected with the fact that certain metal ions were involved in the expression and activation of key enzymes. In addition, it was found that anaerobes in digestion liquid were another potential contributor to the enhanced anaerobic digestion. [Display omitted] [ABSTRACT FROM AUTHOR]

l RL promoted sludge solubilization and SCFA production in the acidogenic phase. l RL increases biochemical methane potential of sludge in the methanogenic phase. l The enhancement of the acidogenic phase by RL is a contributor to the promotion of methane yield. l RL could be applied to anaerobic digestion of waste activated sludge. In this work, a rhamnolipid (RL) pretreatment technology was proposed to promote methane production from two-phase anaerobic digestion of waste activated sludge. In the first phase (i.e., acidogenic phase), the WAS hydrolysis and acidogenesis were significantly enhanced after RL pretreatment for 4 day, under which the concentration of soluble protein and the short-chain fatty acids (SCFA) in the presence of RL at 0.04 g/g TSS was respectively 2.50 and 5.02 times higher than that without RL pretreatment. However, methane production was inhibited in the presence of RL. In the second phase (i.e., methanogenic phase), batch biochemical methane potential tests suggested that the addition of RL is effective in promoting anaerobic methane production. With an increase of RL dosage from 0 to 0.04 g/g TSS, the cumulative methane yield increased from 100.42 ± 3.01 to 168.90 ± 5.42 mL. Although the added RL could be utilized to produce methane, it was not the major contributor to the enhancement of methane yield. Further analysis revealed that total cumulative yield from the entire two-phase anaerobic digestion (sum of the yield of the acidogenic phase and methanogenic phase) increased from 113.42 ± 3.56 to 164.18 ± 5.20 mL when RL dosage increased from 0 to 0.03 g/g TSS, indicating that the addition of RL induced positive effect on the methane production of the entire two-phase anaerobic digestion. The enzyme activity analysis showed that although higher dosages of RL still inhibited the microorganisms related to methanogenesis to some extends in the methanogenic phase, the inhibitory effect was significantly weakened compared to the acidogenic phase. Microbial analysis revealed that RL reduced the abundance of Candidatus_Methanofastidiosum sp. while increased the abundance of Methanosaeta sp., causing the major methanogenesis pathway to change from hydrogenotrophic to aceticlastic. Moreover, the community of hydrolytic microbes and acidogens was shifted in the direction that is conducive to hydrolysis-acidogenesis. The findings reported not only expand the application field of RL, but also may provide supports for sustainable operation of wastewater treatment plants (WWTPs). [Display omitted] [ABSTRACT FROM AUTHOR]

• A granule model embedding metabolisms of DAMO microbes and AnAOB was proposed. • The model predictions matched experimental data from two granular sludge reactors. • The estimated parameter values indicated a stimulated growth of DAMO archaea. • The model managed to simulate the microbial abundance and stratified distribution. • Short HRT and small granule size increased DAMO archaea but reduced DAMO bacteria. The granular bioreactor, characterized by excellent settling velocity, high rate and low cost is an ideal choice for achieving coupled nitrate/nitrite dependent denitrifying anaerobic methane oxidation (n-DAMO) and anaerobic ammonium oxidation (Anammox) process. To fundamentally understand its underlining mechanisms and provide suggestions for process optimization, a granule-based model framework was developed to describe simultaneous anaerobic methane and ammonium oxidation by functional microbes. The proposed model was evaluated based on long-term experimental data from two membrane granular sludge reactors (MGSRs) with different operational conditions. The model possessed of good predictive ability to reproduce removal rates and effluent concentrations of nitrogen species. The predicted biomass abundance in two MGSRs and stratified microbial distribution along granule depth were consistent with experimental observations. The estimated parameter values, with good identifiability and reliability indicated a stimulated growth of DAMO archaea in MGSRs. Both hydraulic retention times (HRTs) and granule sizes have influences on microbial abundance in the MGSR and community distribution inside granules. Within the investigated HRTs from 1.4 h to 20 h and granule sizes from 500 μm to 2900 μm, it was revealed that a proper control of relatively short HRTs and small granule sizes resulted in an increased fraction of DAMO archaea and a reduced DAMO bacteria abundance with AnAOB less impacted, which would lower the required nitrite nitrogen to ammonium nitrogen ratio in the nitritation reactor (prior unit) and thus minimize operational cost in sidestream treatment lines. [ABSTRACT FROM AUTHOR]

Wastewater has been identified as an important carrier for nanoplastics, which could elicit unintended impacts on critical microbial processes. However, the long-term impacts of nanoplastics on anaerobic granular sludge (AGS) for methane recovery from wastewater and the mechanisms involved remains unclear. In this study, we investigated the long term exposure-response relationship between polystyrene nanoplastics (Nano-PS) and AGS. In continuous test over 120 days with 86 days' Nano-PS exposure, feeding wastewater with 10 μg/L of Nano-PS had no significant impacts on the AGS performance. In comparison, higher levels (i.e., 20 and 50 μg/L) of Nano-PS decreased methane production and chemical oxygen demand (COD) removal by 19.0–28.6% and 19.3–30.0%, respectively, along with volatile fatty acids (VFA) accumulation. More extracellular polymeric substance (EPS) was induced by 10 μg/L of Nano-PS as a response to protect microbes, but higher levels (i.e., 20 and 50 μg/L) of Nano-PS decreased EPS generation, causing a decline in granule size and cell viability. Fluorescence tagging found that a large number of Nano-PS agglomerated/accumulated on the outer layer of AGS and even transferred into deeper layers of AGS over exposure time, producing toxic effects to adherent microorganisms, e.g., Longilinea sp., Paludibacter sp. and Methanosaeta sp.. The oxidative stress induced by Nano-PS was revealed to be a key factor for reshaping the AGS, reflected by the increased reactive oxygen species (ROS) generation and lactate dehydrogenase (LDH) release. The sodium dodecyl sulfate (SDS) leached from Nano-PS was also demonstrated to restrain the activities of antioxidant enzymes, thereby further lessening resistance to oxidative stress induced by Nano-PS. This work improves our ability to predict the risks associated with this ubiquitous contaminant in the environment. Image 1 • Long term exposure-response relationship between Nano-PS and AGS is revealed by continuous test. • Nano-PS restrained EPS secretion and AGS activity, resulting in granule breakage. • Nano-PS reduced the abundance of key acidogens and methanogens in AGS. • Nano-PS agglomerated/accumulated on the outer layer of AGS and even transferred into deeper layers of AGS. • Toxic mechanism of Nano-PS to AGS is disclosed. [ABSTRACT FROM AUTHOR]

• Biochar enhanced high-quality biomethane production from PS. • Biochar accelerated PS hydrolysis and enhanced methane production potential of PS. • Biochar incresed buffering capacity, conductivity and alleviated NH 3 inhibition. • Biochar enhanced sludge reduction and reduced sludge disposal costs. • Biochar could be applied to anaerobic digestion of the mixture of WAS and PS. This study conducted batch and continuous tests to reveal the feasibility of corn stover biochar on improving anaerobic digestion of primary sludge (PS). Dosing biochar (1.82, 2.55 and 3.06 g/g Total Solids (TS)) in digester improved methane content increasing from 67.5% to 81.3–87.3% and enhanced methane production by 8.6–17.8%. Model analysis indicated that biochar accelerated PS hydrolysis and enhanced methane potential of PS. The mechanistic studies showed that biochar enhanced process stability provided by strong buffering capacity and alleviated NH 3 inhibition. In continuous test over 116 days, the volatile solids (VS) destruction in the biochar-dosed digester increased by 14.9%, resulting in a 14% reduction in the volume of digestate for disposal. Biochar changed microbial community in an expected direction for anaerobic digestion. This work suggests that biochar technology would apply to co-digestion of WAS and PS to maximize the energy recovery and sludge reduction from the two sludge streams. [ABSTRACT FROM AUTHOR]

• Freezing activated nitrite pretreatment (FN) enhanced anaerobic digestion from WAS. • FN benefited WAS disintegration and the biodegradable of organics released. • FN benefited the dewaterability or pathogens inactivation of digested sludge. • Freezing induced reactive derivatives from nitrite were the major contributors. • FN was economically and environmentally attractive. Nitrite-based pretreatment was demonstrated to effectively improve anaerobic digestion of waste activated sludge. It was found in this work that the freezing activated nitrite pretreatment could further enhance the performances. With the increase of nitrite addition from 0 to 600 mg NO 2 −-N/L during freezing process, the biochemical methane potential of pretreated-sludge gradually increased from 191.3 ± 8.0 to 233.2 ± 10.6 mL per gram volatile solid (VS), while only 178.6 ± 7.3 mL/g VS was obtained in the raw sludge. Mechanism explorations revealed that the freezing activated nitrite pretreatment remarkably facilitated the disintegration of sludge. Excitation emission matrix and fluorescence regional integration analyses further revealed that nitrite addition during freezing process promoted the proportion of biodegradable organics released, thereby providing more bio-available substrates for subsequent anaerobic digestion. Freezing condition induced reactive derivatives from nitrite (e.g., free nitrite acid, NO 2 , N 2 O 3) were assumed to be the major contributors to the enhanced sludge disintegration and recalcitrant organics (e.g., humic acid-like substances) degradation. It was also found that 600 mg NO 2 −-N/L addition activated by freezing pretreatment produced an anaerobically digested sludge with an improved dewaterability, as indicated by the decrease of the specific resistance to filterability and moisture content of dewatered cake. Moreover, 600 mg NO 2 −-N/L addition activated by freezing pretreatment and subsequent anaerobic digestion largely inactivated the pathogens to the levels below Class A biosolids requirements. Considering that nitrite can be in-situ produced in wastewater treatment plants through nitritation of the digestion liquid, this nitrite-based freezing process for sludge pretreatment was environmental-friendly and economically attractive. [ABSTRACT FROM AUTHOR]

Recent investigations verified that calcium peroxide (CaO 2) could be used to pretreat waste activated sludge to promote methane yield from anaerobic digestion. However, the underlying mechanism of how CaO 2 pretreatment promotes methane production is unclear. This work therefore aims to provide insights into such systems. Experimental results showed that with an increase of CaO 2 dosage from 0 to 0.14 g/g VSS (volatile suspended solids) the methane yield increased linearly from 146.3 to 215.9 mL/g VSS. Further increases of CaO 2 resulted in decreases in methane yield. CaO 2 pretreatment promoted the disintegration of sludge and the degradation of sludge recalcitrant organics (especially humus and lignocellulose), thereby providing more substrates for subsequent methane production. Ultraviolet absorption spectroscopy indicated that CaO 2 enhanced the cleavage of unsaturated conjugated bonds and reduced the aromaticity of humus and lignocellulose. Fourier transform infrared spectroscopy showed that CaO 2 changed the structures and functional groups of humus and lignocellulose, making them transform to be biodegradable. GC/MS analyses exhibited that the degradation products of humus and lignocellulose included several types of small molecular organics such as ester-like, acid-like, and alcohol-like substances. Further investigation demonstrated that substantial methane could be produced from these degradation products. It was also found that the presence of recalcitrant organics was detrimental to anaerobes relevant to anaerobic digestion, and the degradation of such recalcitrant organics mitigated their inhibitions to the anaerobes. Model-based analysis suggested that CaO 2 pretreatment increased the maximum methane yield and methane production rate, which were consistent with the analysis above. Image 1 • Pretreatment with 0.14 g/g VSS CaO 2 increased methane yield to 1.3 times from WAS. • CaO 2 pretreatment promoted the degradation of humus and lignocellulose. • CaO 2 could mitigate inhibitions of recalcitrant organics to the anaerobes. • The CaO2 reaction residues inhibited all the microbes relevant to anaerobic digestion. [ABSTRACT FROM AUTHOR]

• Microwave pretreatment (MP) benefited the degradation of PAM. • MP accelerated the release and hydrolysis of organics from PF-WAS. • MP facilitated the breaking of large firm "PAM-sludge" floccules. • Hydrolysis rate of PF-WAS was accelerated through MP. • Biochemical methane potential of PF-WAS was improved through MP. Deterioration of anaerobic digestion can occur with the presence of polyacrylamide (PAM) in waste activated sludge, but the information on alleviating this deterioration is still limited. In this study, the simultaneous alleviation of negative effect of PAM and improvement of methane production during anaerobic digestion was accomplished by microwave pretreatment. Experimental results showed that with the microwave pretreatment times increased from 0 to 12 min, the biochemical methane potential of PAM-flocculated sludge (12 g PAM/kg total solids) asymptotically increased from 123.1 to 242.5 mL/g volatile solids, hydrolysis rate increased from 0.06 to 0.13 d−1. Mechanism analysis indicated that the microwave pretreatment accelerated the release and hydrolysis of organic substrates from PAM-flocculated sludge, facilitated the breaking of large firm "PAM-sludge" floccules, and benefited the degradation of PAM, which alleviated the PAM inhibitory impacts on digestion and meanwhile provided better contact between the released organic substrates and anaerobic bacteria for methane production. [ABSTRACT FROM AUTHOR]