Significant in situ sludge yield reduction in an acidic activated sludge system.

Minimizing sludge generation in activated sludge systems is critical to reducing the operational cost of wastewater treatment plants (WWTPs), particularly for small plants where bioenergy is not recovered. This study introduces a novel acidic activated sludge technology for in situ sludge yield reduction, leveraging acid-tolerant ammonia-oxidizing bacteria (Candidatus Nitrosoglobus). The observed sludge yield (Yobs) was calculated based on the cumulative sludge generation and COD removal during 400 d long-term operation. The acidic process achieved a low Yobs of 0.106 ± 0.004 gMLSS/gCOD at pH 4.6 to 4.8 and in situ free nitrous acid (FNA) of 1 to 3 mg/L, reducing sludge production by 58 % compared to the conventional neutral-pH system (Yobs of 0.250 ± 0.003 gMLSS/gCOD). The acidic system also maintained effective sludge settling and organic matter removal over long-term operation. Mechanism studies revealed that the acidic sludge displayed higher endogenous respiration, sludge hydrolysis rates, and higher soluble microbial products and loosely-bounded extracellular polymer substances, compared to the neutral sludge. It also selectively enriched several hydrolytic genera (e.g., Chryseobacterium, Acidovorax, and Ottowia). Those results indicate that the acidic pH and in situ FNA enhanced sludge disintegration, hydrolysis, and cryptic growth. Besides, a lower intracellular ATP content was observed for acidic sludge than neutral sludge, suggesting potential decoupling of catabolism and anabolism in the acidic sludge. These findings collectively demonstrate that the acidic activated sludge technology could significantly reduce sludge yield, contributing to more cost- and space-effective wastewater management.

Characterization of the redox-active extracellular polymeric substances in an anaerobic methanotrophic consortium.

Anaerobic oxidation of methane (AOM) is a microbial process of importance in the global carbon cycle. AOM is predominantly mediated by anaerobic methanotrophic archaea (ANME), the physiology of which is still poorly understood. Here we present a new addition to the current physiological understanding of ANME by examining, for the first time, the biochemical and redox-active properties of the extracellular polymeric substances (EPS) of an ANME enrichment culture. Using a 'Candidatus Methanoperedens nitroreducens'-dominated methanotrophic consortium as the representative, we found it can produce an EPS matrix featuring a high protein-to-polysaccharide ratio of ∼8. Characterization of EPS using FTIR revealed the dominance of protein-associated amide I and amide II bands in the EPS. XPS characterization revealed the functional group of C-(O/N) from proteins accounted for 63.7% of total carbon. Heme-reactive staining and spectroscopic characterization confirmed the distribution of c-type cytochromes in this protein-dominated EPS, which potentially enabled its electroactive characteristic. Redox-active c-type cytochromes in EPS mediated the EET of 'Ca. M. nitroreducens' for the reduction of Ag+ to metallic Ag, which was confirmed by both ex-situ experiments with extracted soluble EPS and in-situ experiments with pristine EPS matrix surrounding cells. The formation of nanoparticles in the EPS matrix during in-situ extracellular Ag + reduction resulted in a relatively lower intracellular Ag distribution fraction, beneficial for alleviating the Ag toxicity to cells. The results of this study provide the first biochemical information on EPS of anaerobic methanotrophic consortia and a new insight into its physiological role in AOM process.

Wastewater tiling amplicon sequencing in sentinel sites reveals longitudinal dynamics of SARS-CoV-2 variants prevalence.

The ongoing evolution of SARS-CoV-2 is a significant concern, especially with the decrease in clinical sequencing efforts, which impedes the ability of public health sectors to prepare for the emergence of new variants and potential COVID-19 outbreaks. Wastewater-based epidemiology (WBE) has been proposed as a surveillance program to detect and monitor the SARS-CoV-2 variants being transmitted in communities. However, research is limited in evaluating the effectiveness of wastewater collection at sentinel sites for monitoring disease prevalence and variant dynamics, especially in terms of inferring the epidemic patterns on a broader scale, such as at the state/province level. This study utilized a multiplexed tiling amplicon-based sequencing (ATOPlex) to track the longitudinal dynamics of variant of concern (VOC) in wastewater collected from municipalities in Queensland, Australia, spanning from 2020 to 2022. We demonstrated that wastewater epidemiology measured by ATOPlex exhibited a strong and consistent correlation with the number of daily confirmed cases. The VOC dynamics observed in wastewater closely aligned with the dynamic profile reported by clinical sequencing. Wastewater sequencing has the potential to provide early warning information for emerging variants. These findings suggest that WBE at sentinel sites, coupled with sensitive sequencing methods, provides a reliable and long-term disease surveillance strategy.

Superior mainstream partial nitritation in an acidic membrane-aerated biofilm reactor.

Shortcut nitrogen removal holds significant economic appeal for mainstream wastewater treatment. Nevertheless, it is too difficult to achieve the stable suppression of nitrite-oxidizing bacteria (NOB), and simultaneously maintain the activity of ammonia-oxidizing bacteria (AOB). This study proposes to overcome this challenge by employing the novel acid-tolerant AOB, namely "Candidatus Nitrosoglobus", in a membrane-aerated biofilm reactor (MABR). Superior partial nitritation was demonstrated in low-strength wastewater from two aspects. First, the long-term operation (256 days) under the acidic pH range of 5.0 to 5.2 showed the successful NOB washout by the in situ free nitrous acid (FNA) of approximately 1 mg N/L. This was evidenced by the stable nitrite accumulation ratio (NAR) close to 100 % and the disappearance of NOB shown by 16S rRNA gene amplicon sequencing and fluorescence in situ hybridization. Second, oxygen was sufficiently supplied in the MABR, leading to an unprecedentedly high ammonia oxidation rate (AOR) at 2.4 ± 0.1 kg N/(m3 d) at a short hydraulic retention time (HRT) of a mere 30 min. Due to the counter diffusion of substrates, the present acidic MABR displayed a significantly higher apparent oxygen affinity (0.36 ± 0.03 mg O2/L), a marginally lower apparent ammonia affinity (14.9 ± 1.9 mg N/L), and a heightened sensitivity to FNA and pH variations, compared with counterparts determined by flocculant acid-tolerant AOB. Beyond supporting the potential application of shortcut nitrogen removal in mainstream wastewater, this study also offers the attractive prospect of intensifying wastewater treatment by markedly reducing the HRT of the aerobic unit.

Bioreduction of chromate in a syngas-based membrane biofilm reactor.

This study leveraged synthesis gas (syngas), a renewable resource attainable through the gasification of biowaste, to achieve efficient chromate removal from water. To enhance syngas transfer efficiency, a membrane biofilm reactor (MBfR) was employed. Long-term reactor operation showed a stable and high-level chromate removal efficiency > 95%, yielding harmless Cr(III) precipitates, as visualised by scanning electron microscopy and energy dispersive X-ray analysis. Corresponding to the short hydraulic retention time of 0.25 days, a high chromate removal rate of 80 µmol/L/d was attained. In addition to chromate reduction, in situ production of volatile fatty acids (VFAs) by gas fermentation was observed. Three sets of in situ batch tests and two groups of ex situ batch tests jointly unravelled the mechanisms, showing that biological chromate reduction was primarily driven by VFAs produced from in situ syngas fermentation, whereas hydrogen originally present in the syngas played a minor role. 16 S rRNA gene amplicon sequencing has confirmed the enrichment of syngas-fermenting bacteria (such as Sporomusa), who performed in situ gas fermentation leading to the synthesis of VFAs, and organics-utilising bacteria (such as Aquitalea), who utilised VFAs to drive chromate reduction. These findings, combined with batch assays, elucidate the pathways orchestrating synergistic interactions between fermentative microbial cohorts and chromate-reducing microorganisms. The findings facilitate the development of cost-effective strategies for groundwater and drinking water remediation and present an alternative application scenario for syngas.

Ammonium-based bioleaching of toxic metals from sewage sludge in a continuous bioreactor.

The broader reuse of sewage sludge as a soil fertilizer or conditioner is impeded by the presence of toxic metals. Bioleaching, a process that leverages microbial metabolisms and metabolites for metal extraction, is viewed as an economically and environmentally feasible approach for metal removal. This study presents an innovative bioleaching process based on microbial oxidation of ammonia released from sludge hydrolysis, mediated by a novel acid tolerant ammonia-oxidizing bacteria (AOB), Ca. Nitrosoglobus. Over a span of 1024 days, a laboratory-scale bioleaching reactor processing anaerobically digested (AD) sludge achieved an in-situ pH of 2.5 ± 0.3. This acidic environment facilitated efficient leaching of toxic metals from AD sludge, upgrading its quality from Grade C to Grade A (qualified for unrestricted use), according to both stabilization and contaminants criteria. The improved quality of AD sludge could potentially reduce sludge disposal expenses and enable a broader reuse of biosolids. Furthermore, this study revealed a pH-dependent total ammonia affinity of Ca. Nitrosoglobus, with a higher affinity constant at pH 3.5 (67.3 ± 20.7 mg N/L) compared to pH 4.5-7.5 (7.6 - 9.6 mg N/L). This finding indicates that by optimizing ammonium concentrations, the efficiency of this novel ammonium-based bioleaching process could be significantly increased.

Biological bromate reduction coupled with in situ gas fermentation in H2/CO2-based membrane biofilm reactor.

Bromate, a carcinogenic contaminant generated in water disinfection, presents a pressing environmental concern. While biological bromate reduction is an effective remediation approach, its implementation often necessitates the addition of organics, incurring high operational costs. This study demonstrated the efficient biological bromate reduction using H2/CO2 mixture as the feedstock. A membrane biofilm reactor (MBfR) was used for the efficient delivery of gases. Long-term reactor operation showed a high-level bromate removal efficiency of above 95 %, yielding harmless bromide as the final product. Corresponding to the short hydraulic retention time of 0.25 d, a high bromate removal rate of 4 mg Br/L/d was achieved. During the long-term operation, in situ production of volatile fatty acids (VFAs) by gas fermentation was observed, which can be regulated by controlling the gas flow. Three sets of in situ batch tests and two groups of ex situ batch tests jointly unravelled the mechanisms underpinning the efficient bromate removal, showing that the microbial bromate reduction was primarily driven by the VFAs produced from in situ gas fermentation. Microbial community analysis showed an increased abundance of Bacteroidota group from 4.0 % to 18.5 %, which is capable of performing syngas fermentation, and the presence of heterotrophic denitrifiers (e.g., Thauera and Brachymonas), which are known to perform bromate reduction. Together these results for the first time demonstrated the feasibility of using H2/CO2 mixture for bromate removal coupled with in situ VFAs production. The findings can facilitate the development of cost-effective strategies for groundwater and drinking water remediation.

Coupling Partial Nitritation, Anammox and n-DAMO in a membrane aerated biofilm reactor for simultaneous dissolved methane and nitrogen removal.

Anaerobic technologies with downstream autotrophic nitrogen removal have been proposed to enhance bioenergy recovery and transform a wastewater treatment plant from an energy consumer to an energy exporter. However, approximately 20-50 % of the produced methane is dissolved in the anaerobically treated effluent and is easily stripped into the atmosphere in the downstream aerobic process, contributing to the release of greenhouse gas emissions. This study aims to develop a solution to beneficially utilize dissolved methane to support high-level nitrogen removal from anaerobically treated mainstream wastewater. A novel technology, integrating Partial Nitritation, Anammox and Methane-dependent nitrite/nitrate reduction (i.e. PNAM) was demonstrated in a membrane-aerated biofilm reactor (MABR). With the feeding of ∼50 mg NH4+-N/L and ∼20 mg/L dissolved methane at a hydraulic retention time of 15 h, around 90 % of nitrogen and ∼100 % of dissolved methane can be removed together in the MABR. Microbial community characterization revealed that ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB), anammox bacteria, nitrite/nitrate-dependent anaerobic methane oxidation microorganisms (n-DAMO bacteria and archaea) and aerobic methanotrophs co-existed in the established biofilm. Batch tests confirmed the active microbial pathways and showed that AOB, anammox bacteria and n-DAMO microbes were jointly responsible for the nitrogen removal, and dissolved methane was mainly removed by the n-DAMO process, with aerobic methane oxidation making a minor contribution. In addition, the established system was robust against dynamic changes in influent composition. The study provides a promising technology for the simultaneous removal of dissolved methane and nitrogen from domestic wastewater, which can support the transformation of wastewater treatment from an energy- and carbon-intensive process, to one that is energy- and carbon-neutral.

Multi-media interaction improves the efficiency and stability of the bioretention system for stormwater runoff treatment.

Bioretention systems are one of the most widely used stormwater control measures for urban runoff treatment. However, stable and effective dissolved nutrient treatment by bioretention systems is often challenged by complicated stormwater conditions. In this study, pyrite-only (PO), pyrite-biochar (PB), pyrite-woodchip (PW), and pyrite-woodchip-biochar mixed (M) bioretention systems were established to study the feasibility of improving both stability and efficiency in bioretention system via multi-media interaction. PB, PW, and M all showed enhanced dissolved nitrogen and/or phosphorus removal compared to PO, with M demonstrating the highest efficiency and stability under different antecedent drying durations (ADD), pollutant levels, and prolonged precipitation depth. The total dissolved nitrogen and dissolved phosphorus removal in M ranged between 64%-86% and 80%-95%, respectively, with limited organic matter and iron leaching. Pore water, microbial community, and material analysis collectively indicate that pyrite, woodchip, and biochar synergistically facilitated multiple nutrient treatment processes and protected each other against by-product leaching. Pyrite-woodchip interaction greatly increased nitrate removal by facilitating mixotrophic denitrification, while biochar further enhanced ammonium adsorption and expanded the denitrification area. The Fe3+ generated by pyrite aerobic oxidation was adsorbed on the biochar surface and potentially formed a Fe-biochar composite layer, which not only reduced Fe3+-induced pyrite excessive oxidation but also potentially increased organic matter adsorption. Fe (oxyhydr)oxides intermediate product formed by pyrite oxidation, in return, controlled the phosphorus and organic matter leaching from biochar and woodchip. Overall, this study demonstrates that multi-media interaction may enable bioretention systems to achieve stable and effective urban runoff treatment.

Humic substances as electron acceptor for anaerobic oxidation of methane (AOM) and electron shuttle in Mn (IV)-dependent AOM.

Anaerobic methanotrophic archaea (ANME) belonging to the family Methanoperedenaceae are crucial for the global carbon cycle and different biogeochemical processes, owing to their metabolic versatility to couple anaerobic oxidation of methane (AOM) with different electron acceptors. A universal feature of Methanoperedenaceae is the abundant genes encoded in their genomes associated with extracellular electron transfer (EET) pathways. Candidatus. 'Methanoperedens manganicus', an archaeon belonging to the family Methanoperedenaceae, was recently enriched in a bioreactor performing AOM coupled with Mn (IV) reduction. Using this EET-capable ANME, we tested the hypothesis in this study that ANME can catalyse the humic-dependent AOM for growth. A two-year incubation showed that AOM activity can be sustained by Ca. 'M. manganicus' consortium in a bioreactor fed only with humic acids and methane. An isotopic mass balance batch test confirmed that the observed AOM was coupled to the reduction of humic acids. The increase of relative abundance of Ca. 'M. manganicus', and the total archaea population in the microbial community suggested that Ca. 'M. manganicus' can grow on methane and humic acids. The observation of humic-dependent AOM led to a subsequent hypothesis that humic acids could be used as the electron shuttle to mediate the EET in dissimilatory Mn (IV) reduction by Ca. 'M. manganicus'. We tested this hypothesis by adding humic acids to a Ca. 'M. manganicus' dominated-culture, which showed that the AOM rate was doubled by the addition of humic acids. X-ray photoelectron spectroscopy (XPS) showed that quinone moieties were consumed when humic acids worked as electron acceptors while remaining stable when functioning as a shuttle for electron transfer. The results of our study suggest that humic acids may serve as electron shuttles to allow ANME to access more electron acceptors through long-range EET.

Pyrogenic Carbon Promotes Anaerobic Oxidation of Methane Coupled with Iron Reduction via the Redox-Cycling Mechanism.

Pyrogenic carbon (PC) can mediate electron transfer and thus catalyze biogeochemical processes to impact greenhouse gas (GHG) emissions. Here, we demonstrate that PC can contribute to mitigating GHG emissions by promoting the Fe(III)-dependent anaerobic oxidation of methane (AOM). It was found that the amendment PCs in microcosms dominated by Methanoperedenaceae performing Fe(III)-dependent AOM simultaneously promoted the rate of AOM and Fe(III) reduction with a consistent ratio close to the theoretical stoichiometry of 1:8. Further correlation analysis showed that the AOM rate was linearly correlated with the electron exchange capacity, but not the conductivity, of added PC materials, indicating the redox-cycling electron transfer mechanism to promote the Fe(III)-dependent AOM. The mass content of the C═O moiety from differentially treated PCs was well correlated with the AOM rate, suggesting that surface redox-active quinone groups on PCs contribute to facilitating Fe(III)-dependent AOM. Further microbial analyses indicate that PC likely shuttles direct electron transfer from Methanoperedenaceae to Fe(III) reduction. This study provides new insight into the climate-cooling impact of PCs, and our evaluation indicates that the PC-facilitated Fe(III)-dependent AOM could have a significant contribution to suppressing methane emissions from the world's reservoirs.

Challenges of suppressing nitrite-oxidizing bacteria in membrane aerated biofilm reactors by low dissolved oxygen control.

Membrane aerated biofilm reactor (MABR) and shortcut nitrogen removal are two types of solutions to reduce energy consumption in wastewater treatment, with the former improving the aeration efficiency and the latter reducing the oxygen demand. However, integrating these two solutions, i.e., achieving shortcut nitrogen removal in MABR, is challenging due to the difficulty in suppressing nitrite-oxidizing bacteria (NOB). In this study, four MABRs were established to demonstrate the feasibility of initiating, maintaining, and restoring NOB suppression using low dissolved oxygen (DO) control, in the presence and absence of anammox bacteria, respectively. Long-term results revealed that the strict low DO (< 0.1 mg/L) in MABR could initiate and maintain stable NOB suppression for more than five months with nitrite accumulation ratio above 90 %, but it was unable to re-suppress NOB once they prevailed. Moreover, the presence of anammox bacteria increased the threshold of DO level to maintain NOB suppression in MABRs, but it was still incapable to restore the deteriorated NOB suppression in conjunction with low DO control. Mathematical modelling confirmed the experimental results and further explored the differences of NOB suppression in conventional biofilms and MABR biofilms. Simulation results showed that it is more challenging to maintain stable NOB suppression in MABRs compared to conventional biofilms, regardless of biofilm thickness or influent nitrogen concentration. Kinetic mechanisms for NOB suppression in different types of biofilms were proposed, suggesting that it is difficult to wash out NOB developed in the innermost layer of MABR biofilms because of the high oxygen level and low sludge wasting rate. In summary, this study systematically demonstrated the challenges of NOB suppression in MABRs through both experiments and mathematical modelling. These findings provide valuable insights into the applications of MABRs and call for more studies in developing effective strategies to achieve stable shortcut nitrogen removal in this energy-efficient configuration.

Integrated urban water management by coupling iron salt production and application with biogas upgrading.

Integrated urban water management is a well-accepted concept for managing urban water. It requires efficient and integrated technological solutions that enable system-wide gains via a whole-of-system approach. Here, we create a solid link between the manufacturing of an iron salt, its application in an urban water system, and high-quality bioenergy recovery from wastewater. An iron-oxidising electrochemical cell is used to remove CO2 (also H2S and NH3) from biogas, thus achieving biogas upgrading, and simultaneously producing FeCO3. The subsequent dose of the electrochemically produced FeCO3 to wastewater and sludge removes sulfide and phosphate, and enhances sludge settleability and dewaterability, with comparable or superior performance compared to the imported and hazardous iron salts it substitutes (FeCl2, and FeCl3). The process enables water utilities to establish a self-reliant and more secure supply chain to meet its demand for iron salts, at lower economic and environmental costs, and simultaneously achieve recovery of high-quality bioenergy.

Multi-heme cytochrome-mediated extracellular electron transfer by the anaerobic methanotroph 'Candidatus Methanoperedens nitroreducens'.

Anaerobic methanotrophic archaea (ANME) carry out anaerobic oxidation of methane, thus playing a crucial role in the methane cycle. Previous genomic evidence indicates that multi-heme c-type cytochromes (MHCs) may facilitate the extracellular electron transfer (EET) from ANME to different electron sinks. Here, we provide experimental evidence supporting cytochrome-mediated EET for the reduction of metals and electrodes by 'Candidatus Methanoperedens nitroreducens', an ANME acclimated to nitrate reduction. Ferrous iron-targeted fluorescent assays, metatranscriptomics, and single-cell imaging suggest that 'Ca. M. nitroreducens' uses surface-localized redox-active cytochromes for metal reduction. Electrochemical and Raman spectroscopic analyses also support the involvement of c-type cytochrome-mediated EET for electrode reduction. Furthermore, several genes encoding menaquinone cytochrome type-c oxidoreductases and extracellular MHCs are differentially expressed when different electron acceptors are used.

Nitrification in acidic and alkaline environments.

Aerobic nitrification is a key process in the global nitrogen cycle mediated by microorganisms. While nitrification has primarily been studied in near-neutral environments, this process occurs at a wide range of pH values, spanning ecosystems from acidic soils to soda lakes. Aerobic nitrification primarily occurs through the activities of ammonia-oxidising bacteria and archaea, nitrite-oxidising bacteria, and complete ammonia-oxidising (comammox) bacteria adapted to these environments. Here, we review the literature and identify knowledge gaps on the metabolic diversity, ecological distribution, and physiological adaptations of nitrifying microorganisms in acidic and alkaline environments. We emphasise that nitrifying microorganisms depend on a suite of physiological adaptations to maintain pH homeostasis, acquire energy and carbon sources, detoxify reactive nitrogen species, and generate a membrane potential at pH extremes. We also recognize the broader implications of their activities primarily in acidic environments, with a focus on agricultural productivity and nitrous oxide emissions, as well as promising applications in treating municipal wastewater.

Adaptation of anammox process for nitrogen removal from acidic nitritation effluent in a low pH moving bed biofilm reactor.

Acidic partial nitritation (PN) has emerged to be a promisingly stable process in wastewater treatment, which can simultaneously achieve nitrite accumulation and about half of ammonium reduction. However, directly applying anaerobic ammonium oxidation (anammox) process to treat the acidic PN effluent (pH 4-5) is susceptible to the inhibition of anammox bacteria. Here, this study demonstrated the adaptation of anammox process to acidic pH in a moving bed biofilm reactor (MBBR). By feeding the laboratory-scale MBBR with acidic PN effluent (pH = 4.6 ± 0.2), the pH of an anammox reactor was self-sustained in the range of pH 5 - 6. Yet, a high total nitrogen removal efficiency of over 80% at a practical loading rate of up to 149.7 ± 3.9 mg N/L/d was achieved. Comprehensive microbial assessment, including amplicon sequencing, metagenomics, cryosection-FISH, and qPCR, identified that Candidatus Brocadia, close to known neutrophilic members, was the dominant anammox bacteria. Anammox bacteria were found present in the inner layer of thick biofilms but barely present in the surface layer of thick biofilms and in thin biofilms. Results from batch tests also showed that the activity of anammox biofilms could be maintained when subjected to pH 5 at a nitrite concentration of 10 mg N/L, whereas the activity was completely inhibited after disturbing the biofilm structure. These results collectively indicate that the anammox bacteria enriched in the present acidic MBBR could not be inherently acid-tolerant. Instead, the achieved stable anammox performance under the acidic condition is likely due to biofilm stratification and protection. This result highlights the biofilm configuration as a useful solution to address nitrogen removal from acidic PN effluent, and also suggests that biofilm may play a critical role in protecting anammox bacteria found in many acidic nature environments.

Salinity effect on an anaerobic methane- and ammonium-oxidising consortium: Shifts in activity, morphology, osmoregulation and syntrophic relationship.

Nitrate-dependent anaerobic methane oxidation (AOM) is a microbial process of both ecological significance for global methane mitigation and application potential for wastewater treatment. It is mediated by organisms belonging to the archaeal family 'Candidatus Methanoperedenaceae', which have so far mainly been found in freshwater environments. Their potential distribution in saline environments and their physiological responses to salinity variation were still poorly understood. In this study, the responses of the freshwater 'Candidatus Methanoperedens nitroreducens'-dominated consortium to different salinities were investigated using short- and long-term setups. Short-term exposure to salt stress significantly affected nitrate reduction and methane oxidation activities over the tested concentration range of 15-200‰ NaCl, and 'Ca. M. nitroreducens' showed the higher tolerance to high salinity stress than its partner of anammox bacteria. At high salinity concentration, near marine conditions of 37‰, the target organism 'Ca. M. nitroreducens' showed stabilized nitrate reduction activity of 208.5 µmol day-1 gCDW-1 in long-term bioreactors over 300 days, in comparison to 362.9 and 334.3 µmol day-1 gCDW-1 under low-salinity conditions (1.7‰ NaCl) and control conditions (∼15‰ NaCl). Different partners of 'Ca.  M. nitroreducens' evolved in the consortia with three different salinity conditions, suggesting the different syntrophic mechanisms shaped by changes in salinity. A new syntrophic relationship between 'Ca. M. nitroreducens' and Fimicutes and/or Chloroflexi denitrifying populations was identified under the marine salinity condition. Metaproteomic analysis shows that the salinity changes lead to higher expression of response regulators and selective ion (Na+/H+) channeling proteins that can regulate the osmotic pressure between the cell and its environment. The reverse methanogenesis pathway was, however, not impacted. The finding of this study has important implications for the ecological distribution of the nitrate-dependent AOM process in marine environments and the potential of this biotechnological process for the treatment of high-salinity industrial wastewater.

Simultaneous dissolved methane and nitrogen removal from low-strength wastewater using anaerobic granule-based sequencing batch reactor.

Anaerobic treatment of mainstream wastewater has been proposed as a promising solution to enhance bioenergy recovery for wastewater treatment plants (WWTPs). However, the limited organics for downstream nitrogen removal and emissions of dissolved methane into the atmosphere are two major barriers to the broad application of anaerobic wastewater treatment. This study aims to develop a novel technology to overcome these two challenges by achieving simultaneous removal of dissolved methane and nitrogen, and unravel the microbial competitions underpinning the process from the microbial and kinetic perspectives. To this end, a laboratory granule-based sequencing batch reactor (GSBR) coupling anammox and nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) microorganisms was developed to treat wastewater mimicking effluent from mainstream anaerobic treatment. The GSBR achieved high-level nitrogen and dissolved methane removal rates (> 250 mg N/L/d and > 65 mg CH4/L/d) and efficiencies (> 99% total nitrogen removal and > 90% total methane removal) during the long-term demonstration. The availability of different electron acceptors (nitrite or nitrate) imposed significant effects on the removal of ammonium and dissolved methane, as well as on the microbial communities, and the abundance and expression of functional genes. The analysis of apparent microbial kinetics showed that anammox bacteria had a higher nitrite affinity than n-DAMO bacteria, while n-DAMO bacteria had a higher methane affinity than n-DAMO archaea. These kinetics underpin the observation that nitrite is a preferred electron acceptor for removing ammonium and dissolved methane than nitrate. The findings not only extend the applications of novel n-DAMO microorganisms in nitrogen and dissolved methane removal, but also provide insights into microbial cooperation and competition in granular systems.

Novel Use of a Ferric Salt to Enhance Mainstream Nitrogen Removal from Anaerobically Pretreated Wastewater.

This study aims to demonstrate a new technology roadmap to support the ongoing paradigm shift in wastewater management from pollutant removal to resource recovery. This is achieved by developing a novel use of an iron salt (i.e., FeCl3) in an integrated anaerobic wastewater treatment and mainstream anammox process. FeCl3 was chosen to be dosed in a proposed sidestream unit rather than in a primary settler or a mainstream reactor. This causes acidification of returned activated sludge and enables stable suppression of nitrite-oxidizing bacterial activity and excess sludge reduction. A laboratory-scale system, which comprised an anaerobic baffled reactor, a continuous-flow anoxic-aerobic (A/O) reactor, and a secondary settler, was designed to treat real domestic wastewater, with the performance of the system comprehensively monitored under a steady-state condition. The experimental assessments showed that the system had good effluent quality, with total nitrogen and phosphorus concentrations of 12.6 ± 1.3 mg N/L and 0.34 ± 0.05 mg P/L, respectively. It efficiently retained phosphorus in excess sludge (0.18 ± 0.03 g P/g dry sludge), suggesting its potential for further recovery. About half of influent organic carbon was recovered in the form of bioenergy (i.e., methane). This together with low energy consumption revealed that the system could produce a net energy of about 0.11 kWh/m3-wastewater, assessed by an energy balance analysis.

Impact of electrochemically generated iron on the performance of an anaerobic wastewater treatment process.

Anaerobic treatment of domestic wastewater has the advantages of lower biomass yield, lower energy demand and higher energy recover over the conventional aerobic treatment process. However, the anaerobic process has the inherent issues of excessive phosphate and sulfide in effluent and superfluous H2S and CO2 in biogas. An electrochemical method allowing for in-situ generation of Fe2+ in the anode and hydroxide ion (OH-) and H2 in the cathode was proposed to overcome the challenges simultaneously. The effect of electrochemically generated iron (e­iron) on the performance of anaerobic wastewater treatment process was explored with four different dosages in this work. The results showed that compared to control, the experimental system displayed an increase of 13.4-28.4 % in COD removal efficiency, 12.0-21.3 % in CH4 production rate, 79.8-98.5 % in dissolved sulfide reduction, 26.0-96.0 % in phosphate removal efficiency, depending on the e­iron dosage between 40 and 200 mg Fe/L. Dosing of the e­iron significantly upgraded the quality of produced biogas, showing a much lower CO2 and H2S contents in biogas in experimental reactor than that in control reactor. The results thus demonstrated that e­iron can significantly improve the performance of anaerobic wastewater treatment process, bringing multiple benefits with the increase of its dosage regarding effluent and biogas quality.