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The objective of this study was to improve the nitrogen removal efficiency and reduce the start-up period of a single-stage partial nitritation-anammox (SPNA) system using iron particle-integrated anammox granules (IP-IAGs). Anammox granules were enriched in sequencing batch and expanded granular sludge bed (EGSB) reactors. The EGSB reactor produced larger and more uniform granules with higher specific anammox activity. IP-IAGs were then inoculated into a two-stage partial nitritation-anammox reactor treating anaerobic digestion (AD) effluent, followed by an internal recirculation strategy to acclimate the granules to oxygen exposure for SPNA. Finally, the SPNA process operated to treat real AD effluent under optimal conditions of 0.05 L/min aeration intensity (0.01 vvm) and 24 h of hydraulic retention time, achieving TNRE of 86.01 ± 2.64 % and nitrogen removal rate of 0.74 ± 0.04 kg-N/m 3 ·d for 101 d., 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.)

Controllable and recyclable magnetic porous microspheres (MPMs) have been proposed as a means for enhancing the anaerobic digestion (AD) of sludge, as they do not require continuous replenishment and can serve as carriers for anaerobes. However, the effects of MPMs on the interfacial thermodynamics of sludge and the biological responses triggered by abiotic effects in AD systems remain to be clarified. Herein, the underlying mechanisms by which MPMs alter the solid-liquid interface of sludge to drive methanogenesis were investigated. A significant increase in the contents of 13 C and 2 H (D) in methane molecules was observed in the presence of MPMs, suggesting that MPMs might enhance the CO 2 -reduction methanogenesis and participation of water in methane generation. Experimental results demonstrated that the addition of MPMs did not promote the anaerobic bioconversion of soluble organics for methanogenesis, suggesting that the enhanced methanogenesis and water participation were not achieved through promotion of the bioconversion of original liquid-state organics in sludge. Analyses of the capillary force, surface adhesion force, and interfacial proton-coupled electron transfer (PCET) of MPMs revealed that MPMs can enhance mass transfer, effective contact, and electron-proton transfer with sludge. These outcomes were confirmed by the statistical analyses of variations in the interfacial thermodynamics and PCET of sludge with and without MPMs during AD. It was thus proposed that the MPMs enhanced the PCET of sludge and PCET-driven release of protons from water by promoting the interfacial Lewis acid-base interactions of sludge, thereby resulting in the enrichment of free and attached methanogenic consortia and the high energy-conserving metabolic cooperation. This proposition was further confirmed by identifying the predominant syntrophic partners, suggesting that PCET-based efficient methanogenesis was attributable to the enrichment of genomes harbouring CO 2 -reducing pathway and genes encoding water-mediated proton transfer. These findings offer new insights into how substrate properties can be altered by exogenous materials to enable highly efficient methanogenesis., 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.)

The impact of the long-term trace hydrazine (N 2 H 4 ) exogenous supplementation on activity of the anaerobic ammonium oxidation (anammox) biofilm was investigated in a moving bed biofilm reactor (MBBR) for mainstream wastewater treatment. The results of this study demonstrated that the addition of 2-5 mg/L N 2 H 4 enhanced anammox biofilm activity, as evidenced by the augmented nitrogen removal rate (NRR), which increased from 113.4 g/(m 3 ·d) to 126.7 g/(m 3 ·d) with the introduction of 2 mg/L N 2 H 4 . However, a higher concentration of N 2 H 4 (10 mg/L) suppressed anammox activity, leading to a reduced NRR of 91.5 g/(m 3 ·d). Bioindicators revealed that the long-term addition of 2 mg/L N 2 H 4 fostered the accumulation of anammox bacteria (AnAOB) biomass, elevating the volatile suspended solids (VSS) content by 12%. Moreover, the structural composition of extracellular polymeric substances (EPS) within the biofilm was altered, resulting in enhanced biofilm strength within the reactor. The protective mechanism of the biofilm was activated, and EPS secretion was stimulated by the continuous N 2 H 4 supplementation. The introduction of an excess dosage of N 2 H 4 led to alterations in the microbial communities, ultimately resulting in a decline in the performance of the reactor. These findings collectively illustrate that N 2 H 4 , as an intermediate product, can effectively enhance anammox activity within the MBBR for mainstream wastewater treatment. This study contributes to the understanding of the optimization strategies for anammox processes in wastewater treatment systems., 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. Published by Elsevier Ltd.)

The PrrAB two-component system (TCS) is essential for Mycobacterium tuberculosis viability. Previously, it was demonstrated that PrrA binds DNA in the absence of PrrB-mediated transphosphorylation and that non-cognate serine/threonine-kinases phosphorylate PrrA threonine-6 (T6). Therefore, we investigated the differential binding affinity and regulatory properties of the M. tuberculosis-derived wild-type PrrA, PrrA phosphomimetic (D58E, T6E), and PrrA phosphoablative (D58A, T6A) proteins with the prrA Mtb , dosR Mtb , and cydA Mtb genes. While we hypothesized greater DNA binding affinity and more pronounced regulation by PrrA phosphomimetic variants, recombinant, wild-type PrrA Mtb bound DNA with greatest affinity. Collectively, wild-type PrrA Mtb recombinant protein displayed the highest binding affinity to the dosR Mtb promoter (K D 3.46 ± 2.09 nM), followed by the prrA Mtb promoter (K D 9.00 ± 2.66 nM). To establish PrrA Mtb regulatory activity, we constructed M. smegmatis ΔprrAB Msmeg ::prrA Mtb strains with each of the PrrA Mtb variants and extrachromosomal prrA Mtb , dosR Mtb , and cydA Mtb promoter-mCherry reporter fusions. Our findings showed that PrrA Mtb is autoregulatory and induces dosR Mtb expression only during in vitro, hypoxic growth. Combined expression of prrAB Mtb in M. smegmatis ΔprrAB significantly induced cydA Mtb promoter-mCherry expression. Our studies advanced the understanding of PrrA function and PrrAB phosphorylation-mediated regulatory mechanisms and control of mycobacterial dosR and cydA hypoxic and low-oxygen responsive genes., 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.)

Despite being classified as microaerophilic microorganisms, most Campylobacter species can grow anaerobically, using formate or molecular hydrogen (H 2 ) as electron donors, and various nitrogenous and sulfurous compounds as electron acceptors. Herein, we showed that both l-asparagine (l-Asn) and l-aspartic acid (l-Asp) bolster H 2 -driven anaerobic growth in several Campylobacter species, whereas the d-enantiomer form of both asparagine (d-Asn) and aspartic acid (d-Asp) only increased anaerobic growth in Campylobacter concisus strain 13826 and Campylobacter ureolyticus strain NCTC10941. A gene annotated as racD encoding for a putative d/l-Asp racemase was identified in the genome of both strains. Disruption of racD in Cc13826 resulted in the inability of the mutant strain to use either d-enantiomer during anaerobic growth. Hence, our results suggest that the racD gene is required for campylobacters to use either d-Asp or d-Asn. The use of d-Asp by various human opportunistic bacterial pathogens, including C. concisus, C. ureolyticus, and also possibly select strains of Campylobacter gracilis, Campylobacter rectus and Campylobacter showae, is significant, because d-Asp is an important signal molecule for both human nervous and neuroendocrine systems. To our knowledge, this is the first report of pathogens scavenging a d-amino acid essential for human health., Competing Interests: Declaration of competing interest The authors have no conflicts of interest., (Copyright © 2024 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.)

This study evaluated the fate of food delivery boxes when subjected to biological treatments, reproducing at the lab-scale the conditions of full-scale plants. Four paper-based boxes were composted: two made of paper only, one coupled with polylactic acid (PLA), and one with a barrier coating. One paper only box and the box with PLA were also investigated for their anaerobic degradability with biochemical methane potential (BMP) and semi-continuous tests. During composting, the boxes were not recognisable inside the compost after four (paper only boxes), eight (box with PLA), and twelve (box with barrier coating) weeks. In BMP tests, the paper only box showed a degradability similar to that of food waste (92 %), while the box with PLA degraded only at 76 %. Furthermore, undigested pieces of PLA were found in semi-continuous tests. Accordingly, paper resulted suitable for biological treatments, while the presence of PLA or other barrier coatings can be critical., 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 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Variation of dissolved organic matters (DOM) in mainstream anammox process has received limited attention. This study systematically characterized DOM and dissolved organic nitrogen (DON) in a full-scale mainstream anammox wastewater treatment plant (WWTP) using spectroscopy and Fourier transform-ion cyclotron resonance mass spectrometry. Roles of bacterial community structures related with temperatures on DOM and DON transformations were analyzed. Results indicated that the WWTP removed highly bioavailable, S-containing DOM while producing more unsaturated, aromatic, and N-containing DOM. Higher relative abundances of Proteobacteria and Chloroflexi at low temperature resulted in greater removal rates of proteins, SMP-like and humic acid-like substances. At high temperature, higher relative abundance of Actinobacteriota increased lignin production. Principal component analysis revealed that temperature significantly impacted DOM characteristics compared to DON. These findings are crucial for understanding DOM and DON transformation during mainstream anammox WWTP., 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.)

The long-term performance of anaerobic digestion (AD) often decreases substantially when treating swine wastewater contaminated with heavy metals. However, the toxicological characteristics and mechanisms of continuous exposure to heavy metals under different organic loading rates (OLR) are still poorly understood. In these semi-continuous AD experiments, it was found that zinc concentrations of 40 mg/L only deteriorated the reductive environments of AD. In comparison, a concentration of 2.0 mg/L probably facilitated the reproduction of microorganisms in the operating digesters with a constant OLR of 0.51 g COD/(L·d). Nevertheless, when the OLR was increased to 2.30 g COD/(L·d), 2.0 mg/L zinc inhibited various life activities of microorganisms at the molecular level within only 10 days. Hence, even though 2.0 mg/L zinc could promote AD performances from a macroscopic perspective, it had potential inhibitory effects on AD. Therefore, this study deepens the understanding of the inhibitions caused by heavy metals on AD and the metabolic laws of anaerobic microorganisms in swine wastewater treatment. These results could be referred to for enhancing AD in the presence of zinc in practical swine wastewater treatment., 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.)

Anaerobically-treated palm oil mill effluent (POME) still has unacceptable properties for water recycling and reuse, with an unpleasant appearance due to the brownish color caused by tannins and phenolic compounds. This study proposes an approach for treating anaerobically-treated POME for water recycling by combining organic precipitation, electrocoagulation (EC), and ion-exchange resin, followed by reverse osmosis (RO) membrane filtration in series. The results indicated that the organic precipitation enhanced the efficiency of EC treatment in reducing the concentrations of tannins, color, and chemical oxygen demand (COD) of the anaerobically-treated POME effluent, with reductions of 95.73%, 96.31%, and 93.96% for tannin, color, and COD, respectively. Moreover, organic precipitation affected the effectiveness of Ca 2+ and Mg 2+ ion removal using ion exchange resin and RO membrane filtration. Without prior organic precipitation, the ion-exchange resin process required a longer contact time, and the RO membrane filtration treatment was hardly effective in removing total dissolved solids (TDS). The combined process gave a water quality that meets the criteria set by the Thailand Ministry of Industry for industrial boiler use (COD 88 mg/L, TDS <0.001 mg/L, water hardness <5 mg-CaCO 3 /L, and pH 6.9)., 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.)

The impact of weak magnetic field (WMF) on anaerobic digestion (AD) performance of waste activated sludge (WAS) and underlying mechanism were investigated. Results showed that WMF significantly stimulated the methane yield by 12.9∼25.1% with 15 and 30 mT WMF addition, but high WMF (60 mT) attenuated the positive effect. The WMF enriched the anaerobic microbes, especially the acetoclastic and hydrogenotrophic methanogen. Additionally, the WMF dramatically facilitated the metabolic pathways of key enzymes for methanogenesis, which was validated by the significant increase of absolute abundance of anaerobic functional genes (mcrA). The enzyme activities of ATP and F420 were also significantly promoted by 30 mT WMF, but high WMF (60 mT) resulted in increased activity of lactate dehydrogenase. This study reveals that low WMF can promote AD performance of WAS through enhancing microbial activities especially methanogen, but high WMF leads to the loss of cell membrane integrity and attenuates its positive effect., 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.)

The generation of food waste (FW) is increasing at an alarming rate, contributing to a total of 32% of all the waste produced globally. Anaerobic digestion (AD) is an effective method for dealing with organic wastes of various compositions, like FW. Waste valorization into value-added products has increased due to the conversion of FW into biogas using AD technology. A variety of pathways are adopted by microbes to avoid unfavorable conditions in AD, including competition between sulfate-reducing bacteria and methane (CH 4 )-forming bacteria. Anaerobic bacteria decompose organic matter to produce biogas, a digester gas. The composition depends on the type of raw material and the method by which the digestion process is conducted. Studies have shown that the biogas produced by AD contains 65-75% CH 4 and 35-45% carbon dioxide (CO 2 ). Methanothrix soehngenii and Methanosaeta concilii are examples of species that convert acetate to CH 4 and CO 2 . Methanobacterium bryantii , Methanobacterium thermoautotrophicum , and Methanobrevibacter arboriphilus are examples of species that produce CH 4 from hydrogen and CO 2 . Methanobacterium formicicum , Methanobrevibacter smithii , and Methanococcus voltae are examples of species that consume formate, hydrogen, and CO 2 and produce CH 4 . The popularity of AD has increased for the development of biorefinery because it is seen as a more environmentally acceptable alternative in comparison to physico-chemical techniques for resource and energy recovery. The review examines the possibility of using accessible FW to produce important value-added products such as organic acids (acetate/butyrate), biopolymers, and other essential value-added products.

Because of the naturally limited anaerobic degradability and limited biogas yield of raw sludge (RS), this study aims to increase the biogas production of primary sludge (PS) and waste activated sludge (WAS) by the integration of thermal alkaline process (TAP). PH 11 is confirmed to be the most suitable pH value for the TAP of both sludges. Moreover, with the pretreatment at pH 11 and 160 °C (6 bar) for 30 min, the investigated PSs and WASs achieved an increased biogas production of up to 81 % and 72 %, respectively. The improved net electricity production of WASs after TAP varied between 15-43 % compared to conventional WAS digestion. However, the TAP of PS at pH 11 enhanced the biogas production by 1-81 %, which did not constantly contribute to an improved net electricity production., 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. Published by Elsevier Ltd.)

A pilot plant with a conventional activated sludge (CAS) system with intermittent aeration (IA) was monitored. The system was configured as an Oxic Settling Anaerobic (OSA) process with the insertion of one anaerobic side-stream reactor (ASSR). The pilot plant was fed with real wastewater and an intensive experimental campaign was carried out including sludge minimization, nitrogen and carbon removal, GHG emissions and biokinetic parameters. The experimental campaign was divided into periods: Period I, II, and III. In Periods I and II, the ASSR reactor was operated with two different hydraulic retention times (HRT), 4 and 6 h, with an aeration/non-aeration ratio of 30 min/30 min. In Period III, the HRT in the anaerobic reactor was the same as in Period II. In contrast, the biological reactor's aerated/non-aerated ratio was increased to 40 min/20 min. Results demonstrated that combining IA and OSA might be effective in the reduction of excess sludge production. The yield coefficient decreased from Period I to Period II (Y obs from 0.41 to 0.25 gTSS gCOD -1 , in Period I and II, respectively). Nevertheless, the HRT increase in the ASSR compromised the system performance regarding nitrification and greenhouse gas emissions and worsened the sludge settleability. However, the increase in the aeration duration was beneficial in restoring the system's nitrification and denitrification ability and carbon footprint. The lowest carbon footprint was obtained during Period III (6.8 kgCO 2 /d)., 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 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The anaerobic digestion (AD) process is one of the most practiced technologies for the remediation of organic waste and maximization of energy recovery in terms of biogas or biomethane. The presence of other gaseous components in biogas, e.g. CO 2 and H 2 S, often makes its direct application in engines and electricity production unsuitable. This work aimed to develop and utilize an algae-assisted microbial fuel cell (AMFC) for the purification of biogas by removing both CO 2 and H 2 S and simultaneous bioelectricity generation. In addition to biogas clean-up, elemental sulfur recovery and CO 2 utilization for algae cultivation add value to the proposed AMFC process. Experiments were performed with both sulfide and bicarbonate in their dissolved form, in the respective anodic and cathodic chambers of the AMFC. The sulfide concentration was varied from 100 to 800 mg/l and the AMFC exhibited a sulfide removal efficiency exceeding 97% at all concentrations tested. The process efficiency dropped, however, at sulfide concentrations above 300 mg/l in terms of both sulfide removal and power output. The AMFC performed best at 400 mg/l sulfide by exhibiting a power density of 24.99 mW/m 3 and sulfide removal efficiency of 98.87%. The system exhibited columbic efficiency (CE %) in the range of 7.85-80%. The total alkalinity representing CO 2 , carbonate and bicarbonate levels in the algae-based system was reduced by 49.54%. The electrical energy recovered from the AMFC was 0.1 kWh/m 3 and the total energy recovery, which is the sum of the electrical and algal lipid energy, amounted to 7.25 kWh/m 3 .

Greenhouse gas emissions from liquid livestock manure storage significantly contribute to global warming. Accurate farm-scale models are essential for predicting these emissions and evaluating manure management strategies, but they rely on multiple parameters describing carbon loss dynamics. Surface respiration may significantly influence carbon loss and methane emission, yet it is not explicitly included in current models. We conducted experiments to measure pig manure surface respiration rate and its effect on organic matter degradation and methane and carbon dioxide emissions. Manure was incubated for 283 days at 10°C or 20°C under aerobic or anaerobic conditions, while measuring methane and carbon dioxide emission. This was followed by anaerobic digestion at 38°C. Surface respiration reduced the organic matter content, and the effect was temperature dependent. Methane emission was not affected by surface respiration, suggesting that substrate availability was not rate-limiting for methanogenesis. Surface respiration rates were 18.1 ± 3.5 g CO 2 m -2 day -1 at 10°C and 37.1 ± 13.1 g CO 2 m -2 day -1 at 20°C (mean ± standard deviation) and were consistent with microsensor measurements of oxygen consumption in different manure surfaces. Based on these results, temperature- and surface area-dependent respiration was incorporated in the existing anaerobic biodegradation model (ABM). Simulations showed that surface respiration accounts for 29% of carbon losses in a typical pig house and 8% for outdoor storage. Developing and refining algorithms for diverse carbon transformations, such as surface respiration, is crucial for evaluating the potential for methane emission and identification of variables that control emissions at the farm scale., (© 2024 The Author(s). Journal of Environmental Quality published by Wiley Periodicals LLC on behalf of American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America.)

This study investigated the effects of varying lipid ratios on the anaerobic co-digestion of high-lipid food waste (FW) in a mesophilic anaerobic membrane bioreactor (AnMBR). At a lipid concentration of 5 %, optimal biogas production (3.84 L/L/d) and lipid removal efficiency (78 %) were achieved; however, increasing lipid concentrations resulted in significant accumulations of long-chain fatty acids (LCFAs) and volatile fatty acids (VFAs). Batch tests further demonstrated the impact of various types of LCFAs, with stearic acid showing the slowest microbial growth rate (0.033d -1 ), confirming its role in the accumulation of acetate-dominated VFAs, potentially limiting the methanogenesis process at elevated lipid levels. Furthermore, at 8 % lipid content, the downregulation of key LCFA degradation enzymes and dominance of hydrogenotrophic methanogens indicated adverse conditions. The importance of the intricate interplay between LCFA degradation kinetics and microbial community for the system efficiency was evidenced, offering insights for optimizing and managing high-lipidic wastes., 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.)

Resource depletion and climate changes due to human activities and excessive burning of fossil fuels are the driving forces to explore alternatives clean energy resources. The objective of this study was to investigate the potential of potato peel waste (PPW) at various temperatures T15 (15 °C), T25 (25 °C), and T35 (35 °C) in anaerobic digestion (AD) for biogas generation. The highest biogas and CH 4 production (117 mL VS -g and 74 mL VS -g ) was observed by applying 35 °C (T35) as compared with T25 (65 mL VS -g and 22 mL VS -g ) on day 6. Changes in microbial diversity associated with different temperatures were also explored. The Shannon index of bacterial community was not significantly affected, while there was a positive correlation of archaeal community with the applied temperatures. The bacterial phyla Firmicutes were strongly affected by T35 (39%), whereas Lactobacillus was the dominant genera at T15 (27%). Methanobacterium and Methanosarcina, as archaeal genera, dominated in T35 temperature reactors. In brief, at T35, Proteiniphilum and Methanosarcina were positively correlated with volatile fatty acids (VFAs) concentration. Spearman correlation revealed dynamic interspecies interactions among bacterial and archaeal genera; facilitating the AD system. This study revealed that temperature variations can enhance the microbial community of the AD system, leading to increased biogas production. It is recommended for optimizing the AD of food wastes., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Stillage of American whiskey (e.g., bourbon) manufacturing is an abundant byproduct that is distinguished from fuel ethanol and malt whisky stillage materials by its highly inconsistent nature due to variability in mash bill composition. The impact of stillage physicochemical characteristics on biomethane production through anaerobic digestion was assessed by characterizing American whiskey stillage samples before batch biochemical methane potential tests of whole stillage. A maximum methane yield of 419 Nml CH 4 /g VS was obtained under food to microbe ratio (F: M) of 0.5 and organic loading rate (OLR) of 10 g VS/L while digester instability was noted under F: M ratios exceeding 0.5 under batch production. Methane production was significantly influenced by the mash bill composition with lowest methane yields obtained with higher rye content (rye whiskey) and highest methane yields obtained with higher corn content (bourbon or corn whiskey). A multiple linear regression model including C, P, N, and Na was able to accurately describe the methane yield (R 2  = 0.93). This study provides valuable insights to aid the design of anaerobic digesters generating renewable natural gas from heterogeneous American whiskey stillage., 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. Published by Elsevier Ltd.)

A novel bio-carrier, PVA/Fe/starch gel bead, was fabricated and developed to enhance the anaerobic treatment performance of synthetic azo dye-containing wastewater. PVA-gel beads with 5 % magnetite and 0.5 % starch were optimal for physical strength and treatment performance. A pair of 2 L-up-flow anaerobic sludge blankets (UASB), one with the bead (UB) and another without (U) as a controller, operated continuously at 30 °C and an HRT of 11-24 h for 302 days. UB showed better performance than U in most phases, especially with influent dye of 200 mg·L -1 , suggesting a greater tolerance to dye toxicity of UB than U. Microbial analysis revealed that the PVA/Fe/starch gel beads successfully captured the dye degrader Clostridium. Diversity indices indicated that PVA/Fe/Starch gel beads effectively support microbial diversity and resilience under varying dye concentrations. Overall, these findings demonstrate the potential of PVA/Fe/Starch gel beads to improve the stability and efficiency of the dye treatment system., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Takahiro Watari reports financial support was provided by Japan Society for the Promotion of Science. Takahiro Watari reports financial support was provided by Kurita Water and Environment Foundation. If there are other authors, they 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.)

Energy self-circulation systems, defined as energy systems incorporating the recycling utilization of waste biomass, have been proposed to reduce greenhouse gases emissions from livestock sector. In this study, a comprehensive review of the situation and challenges of biogas utilization in the livestock industry was provided. Moreover, two technological routes were proposed for a circular livestock system combined with Powerfuels production (CP-CLS), starting from biogas combustion for power generation and steam reforming to the sustainable development path of synthesizing, storing, and utilizing Powerfuels. The self-circulation capability and comprehensive benefits of the CP-CLS life cycle was discussed, along with future application scenarios and proposed standards for Powerfuels. To realize this potential, continuous research, development, and policy support are crucial. This study envisions the next generation of energy self-circulation systems, which expects to reduce the negative effect of livestock industry on climate change and promote sustainable development., 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.)

For revealing the influence of temperature on volatile fatty acids (VFAs) generation from primary sludge (PS) during the anaerobic fermentation process facilitated by peroxymonosulfate (PMS), five fermentation groups (15, 25, 35, 45, and 55 °C) were designed. The results indicated that the production of VFAs (5148 mg COD/L) and acetic acid (2019 mg COD/L) reached their peaks at 45 °C. High-throughput sequencing technology disclosed that Firmicutes, Proteobacteria, and Actinobacteria was the dominant phyla, carbohydrate metabolism and membrane transport were the most vigorous at 45 °C. Additionally, higher temperature and PMS exhibit synergistic effects in promoting VFAs accumulation. This study unveiled the mechanism of the effect of the pretreatment of PS with PMS on the VFAs production, which established a theoretical foundation for the production of VFAs., 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.)

The anaerobic biological treatment of landfill leachate frequently encounters the souring problems because of the high concentration of organic in landfill leachate. Nonetheless, the performance of anaerobic membrane bioreactor (AnMBR) is commendable in terms of removal of organic compounds. Hence, this study explored the effect of organic concentration and hydraulic retention time(HRT) on the removal performance of actual landfill leachate, additionally, carbon conversion through carbon mass balance analysis was analyzed, in order to determine the optimal treatment potential of AnMBR in treating landfill leachate. For HRT values between 14.5 h and 34.6 h, and the influent COD (Chemical Oxygen Demand) range of 12,773.33-15706.67 mg/L, AnMBR could efficiently treat landfill leachate. As HRT was fixed at 14.5 h and influent COD was around 12,206.7-15,373.33 mg/L, AnMBR achieved a maximum organic removal rate of 18.22 ± 0.51 kg COD/(m 3 ∙d) with methane yield of 0.24 ± 0.01 m 3 CH 4 /kg COD and methane content of 88.26%. Based on carbon mass balance, increasing COD concentration in the influent (less than 16,000 mg/L) boosted the conversion of organic compounds (45.19 ± 4.24%) into CH 4 ; while decreasing HRT (more than 27.0 h) also promoted the conversion of organic compounds into CH 4 (38.36-60.93%) resulting in a decreased TOC (Total Organic Carbon) loss by 2.02-7.19% with outflow. AnMBR may efficiently produce methane while treating landfill leachate by assessing the random forest model (RF) and adjusting the balance between HRT and influent COD concentration., 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.)

An iron-retrofitted anaerobic baffled reactor (ABR) system was developed for the effective treatment of rural wastewater with reduced maintenance demand and aeration costs. Average removal efficiencies of chemical oxygen demand, total nitrogen and total phosphorus of 99.4%, 62.7% and 92.6% were achieved respectively, when the ABR system was operating at steady state. With effective bioreduction of Fe III in the anaerobic chambers, phosphorus was immobilized in the sludge as vivianite, the sole phosphorus-carrying mineral. The Fe III in the recirculated sludge induced Feammox in the ABR reactor, contributing 14.8% to total nitrogen removal. Biophase separation and enrichment of microorganisms associated with iron and nitrogen transformations were observed in the system after Fe dosing, which enhanced the removal of pollutants. The coupling of Feammox and vivianite crystallization to remove nitrogen and phosphorus in an iron-retrofitted ABR would appear to be a promising technology for rural wastewater treatment., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Xiang Cheng reports financial support was provided by National Key Research and Development Program of China. Xiang Cheng reports financial support was provided by Natural Science Foundation of China. If there are other authors, they 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.)

Considering the high organic matter contents and pollutants in sewage sludge (SS) and food waste (FW), seeking green and effective technology for energy recovery and pollutant control is a big challenge. In this study, we proposed a integrated technology combing SS mass separation by hydrothermal pretreatment, methane production from co-digestion of hydrothermally treated sewage sludge (HSS) centrate and FW, and biochar production from co-pyrolysis of HSS cake and digestate with heavy metal immobilization for synergistic utilization of SS and FW. The results showed that the co-digestion of HSS centrate with FW reduced the NH 4 + -N concentration and promoted volatile fatty acids conversion, leading to a more robust anaerobic system for better methane generation. Among the co-pyrolysis of HSS cake and digestate, digestate addition improved biochar quality with heavy metals immobilization and toxicity reduction. Following the lab-scale investigation, the pilot-scale verification was successfully performed (except the co-digestion process). The mass and energy balance revealed that the produced methane could supply the whole energy consumption of the integrated system with 26.2 t biochar generation for treating 300 t SS and 120 t FW. This study presents a new strategy and technology validation for synergistic treatment of SS and FW with resource recovery and pollutants control., 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. Published by Elsevier Ltd.)

The organic loading rate (OLR) is a critical parameter affecting the stability of dry anaerobic digestion (AD) of kitchen waste (KW), and significantly impacting the variations in physicochemical parameters and microbial communities. However, the evolution of quorum sensing (QS) and their role on anaerobic biochemical metabolism during the increase in OLR in dry AD remain unknown. Therefore, this study systematically elucidated the matter through multi-omics analysis based on a pilot-scale dry AD of KW. The results demonstrated that fluctuations in the OLR significantly influenced the microbial QS in dry AD. When the OLR ≤4.0 g·VS/L·d, the system operated stably, and methane production increased. The enrichment of Proteobacteria was crucial for sustaining high levels of functional genes associated with various types of QS, including acyl-homoserine lactones (AI-1), autoinducer-2 (AI-2), autoinducer-3 (AI-3), and gamma-aminobutyric acid (GABA). This enabled cooperative communication among microbes under low OLR. Furthermore, most genes associated with these QS processes positively affected hydrolysis, acidogenesis, and methanogenesis. When the OLR increased to 6.0 g·VS/L·d, the fatty acids and hydrogen partial pressure increased significantly. The autoinducing peptides (AIP)-type became the predominant QS and was positively correlated with fatty acids abundance. Syntrophaceticus and Syntrophomonas may promote syntrophic oxidation of acetate at high OLR through AIP-type QS. These findings provided new insights into the QS processes of microbes during dry AD of KW and a theoretical foundation for optimizing biochemical metabolic processes in dry AD through QS., 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.)

Food waste (FW) and its biogas residue were considered as sources of terrestrial microplastics (MPs) and phthalic acid esters (PAEs) contamination. However, there was a lack of research and understanding of the MPs and PAEs pollution problem in FW dry anaerobic digestion process (DADP). The MPs and PAEs in three stages of the DADP with the largest monomer disposal scale in China were identified. At the biogas residue extrusion stage, MPs abundance and PAEs concentration reached the highest values, which were 3.63 ± 0.45 × 10 3 N·kg -1 and 3.62 ± 0.72 mg·kg -1 , respectively. Furthermore, there was a significant positive correlation between MPs and PAEs throughout the process (p < 0.05). Although bacteria and fungi with plastic degradation potential were present in all stages, the contamination problem of MPs and PAEs cannot be completely solved through DADP. This study provides a scientific basis for preventing and controlling the pollution of MPs and PAEs., 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.)

Anammox is recognized as a prospective alternative for future biological nitrogen removal technologies. However, the nitrate by-products produced by anammox bacteria limit its overall nitrogen removal efficiency below 88 %. This study introduced Fe(III) into the anammox bioreactor to enhance the nitrogen removal efficiency to approximately 95 %, surpassing the biochemical limit of 88 % imposed by anammox stoichiometry. Anammox sludge was demonstrated to utilize extracellular polymeric substances to reduce Fe(III) into Fe(II), and this process promoted the dominance of Ca. Brocadia. The iron addition improved the abundance of narGHI genes and facilitated the partial dissimilatory nitrate reduction to ammonium, with nitrite as the end product. The accumulated nitrite was then eliminated through the anammox pathway, along with the excess ammonium (30 mg/L) in the influent. Overall, this study deepens our understanding of the enhanced nitrogen removal triggered by Fe(III) in anammox sludge and offers an effective approach to boost anammox process., 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. Published by Elsevier Ltd.)

During in situ biomethanation, microbial communities can convert complex Organic Matter (OM) and H 2 into CH 4 . OM biodegradation was compared between Anaerobic Digestion (AD) and in situ biomethanation, in semi-continuous processes, using two inocula from the digester (D) and the post-digester (PoD) of an AD plant. The impact of H 2 on OM degradation was assessed using a fractionation method. Operational parameters included 20 days of hydraulic retention time and 1.5 g VS .L -1 .d -1 of organic loading rate. During in situ biomethanation, 485 NmL of H 2 were injected for each feeding (3 times a week). Maximum organic COD removal was 0.6 gCOD in AD control and at least 1.6 gCOD for in situ biomethanation. Therefore, COD removal was 2.5 times higher with H 2 injections. These results bring out the potential of H 2 injections during AD, not only for CO 2 consumption but also for better OM degradation., 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 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Quorum sensing (QS) plays an important role in the social behavior of microbial communities. Anaerobic digestion (AD) is a biological process using anaerobic microorganisms to degrade organic macromolecules into small molecules for biogas and biofertilizer production. In AD, the QS signaling molecule N-acyl homoserine lactones (AHLs) induces bacterial metabolism, improving AD process efficiency. However, there are fewer systematic reports about QS regulation of microbial behavior in AD. In this report, the effects of signaling molecules on extracellular polymer secretion, biofilm formation, granulation of granular sludge and bacterial metabolism in AD were investigated in detail. At present, the regulation behavior of QS on AD is a group phenomenon, and there are few in-depth studies on the regulation pathway. Therefore, we conducted an in-depth analysis of the pure culture system, granular sludge and reactor in the AD. Then we pointed out that the future application potential of QS in the AD may be combined with quorum quenching (QQ) and omics technology, which is of great significance for the future application of AD., 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. Published by Elsevier Ltd.)

Rare earth elements result in substantial tailings wastewater with high ammonium and nitrate during extraction. In this study, a temperature-resilient Anammox process was employed for efficient treatment of rare earth element tailings wastewater through implementing synergistic nitrite supply by partial nitritation (PN) and partial denitrification (PD). Enhancing temperature resilience of Anammox process relies on dynamic management of DO and COD inputs to shift the dominant nitrite supplier from PN to PD, stable PD (NAR ≥ 90 %) can boost nitrogen removal by Anammox to 97.8 %. The nitrogen removal rate and nitrogen removal efficiency at 10.6 °C could maintain at 0.12 kgN/m 3 ·d -1 and 92.5 %, respectively. Microbial analysis reveals that Nitrosomonas, Thauera, and Candidatus&#95;Kuenenia are the predominant genera responsible for nitrite supply and nitrogen removal, localized within the gas channels of granules, flocs, and micro-granules, respectively. Keeping the influent C/NO 3 - -N ratio below 1.7 is ideal to prevent overgrowth of Thauera and maintain system stability., 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.)

Despite the potential of sodium sulfide (Na 2 S) for phosphorus (P) recovery from iron-phosphate waste, the underlying mechanism regarding its impact on P conversion and product quality has not been well addressed. In this study, the effects of Na 2 S addition on P release and recovery from a chemical-enhanced phosphorus removal (CEPR) sludge during anaerobic fermentation were systematically investigated. The results revealed that the effective mobilization of P bound to Fe (Fe-P) by Na 2 S dominated the massive P release from the CEPR sludge, while the organic P (OP) release was not significantly enhanced during anaerobic fermentation. Due to the rapid reaction of Na 2 S with Fe-P and the prevention of Fe(II)-P precipitation by excess S 2- , the Fe-P was decreased by 9.7%, 15.2% and 24.9% at S:Fe molar ratios of 0.3, 0.5 and 1, respectively. After anaerobic fermentation, the released P mainly existed as soluble phosphate (SP), P bound to Ca (Ca-P) and P bound to Al (Al-P). The nitrogen and P contents in the fermentation supernatant significantly increased with higher S:Fe ratios, facilitating the efficient recovery of P as high-purity struvite. However, the increased Na 2 S dosage deteriorated the sludge dewaterability because of the dissolution of hydrophilic extracellular polymeric substances and the looser secondary structure of proteins. Comprehensively considering the P recovery, sludge dewaterability and economic cost, the optimal Na 2 S dosage was determined at the S:Fe ratio of 0.3. These findings provide novel insights into the role of Na 2 S in P recovery as struvite from CEPR sludge., 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.)

Construction and demolition wastes (CDWs) have become a significant environmental concern due to urbanization. CDWs in landfill sites can generate high-pH leachate and various constituents (e.g., acetate and sulfate) following the dissolution of cement material, which may affect subsurface biogeochemical properties. However, the impact of CDW leachate on microbial reactions and community compositions in subsurface environments remains unclear. Therefore, we created columns composed of layers of concrete debris containing-soil (CDS) and underlying CDW-free soil, and fed them artificial groundwater with or without acetate and/or sulfate. In all columns, the initial pH 5.6 of the underlying soil layer rapidly increased to 10.8 (without acetate and sulfate), 10.1 (with sulfate), 10.1 (with acetate), and 8.3 (with acetate and sulfate) within 35 days. Alkaliphilic or alkaline-resistant microbes including Hydrogenophaga, Silanimonas, Algoriphagus, and/or Dethiobacter were dominant throughout the incubation in all columns, and their relative abundance was highest in the column without acetate and sulfate (50.7-86.6%). Fe(III) and sulfate reduction did not occur in the underlying soil layer without acetate. However, in the column with acetate alone, pH was decreased to 9.9 after day 85 and Fe(II) was produced with an increase in the relative abundance of Fe(III)-reducing bacteria up to 9.1%, followed by an increase in the methanogenic archaea Methanosarcina, suggestive of methanogenesis. In the column with both acetate and sulfate, Fe(III) and sulfate reduction occurred along with an increase in both Fe(III)- and sulfate-reducing bacteria (19.1 and 17.7%, respectively), while Methanosarcina appeared later. The results demonstrate that microbial Fe(III)- and sulfate-reduction and acetoclastic methanogenesis can occur even in soils with highly alkaline pH resulting from the dissolution of concrete debris., 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.)

An effective strategy for utilizing anaerobic digestates is required to promote biomass power generation. We developed an anaerobic digestate tablet using liquid dairy cattle manure derived from a small mesophilic anaerobic digester installed on a dairy farm. Anaerobic digestate tablets are intended for use in the fertilization of oligotrophic coastal seas to promote primary production. The purpose of this study was to evaluate (1) the dissolution behavior of nutrients from anaerobic digestate tablets and (2) the effect of the application of anaerobic digestate tablets on the growth of benthic microalgae using a culture experiment. Batch experiments were conducted to investigate the dissolution behavior of the nutrients. Cumulative amounts of dissolved inorganic nitrogen and phosphate in the anaerobic digestate tablet ranged from 110 to 28.9 μg g -1 after 28 days. The dissolved inorganic nitrogen in the anaerobic digestate tablet was mainly ammonium nitrogen and accounted for 92.4-96.9%, which is advantageous for the growth of microalgae. The growth curve of the benthic microalga Nitzchia longissima was monitored using f/2 medium added to the anaerobic digestate tablet. The growth of Nitzchia longissima was two orders of magnitude greater than that of the positive control. The enhanced growth of Nitzchia longissima by the anaerobic digestate tablet was considered a concomitant effect of moderate dissolution of ammonium nitrogen and high affinity for benthic microalgae. In conclusion, the anaerobic digestate tablets prepared in this study have the advantage of supplying nitrogen to benthic microalgae. This study proposes a new method for utilizing anaerobic digestates., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The conversion of carbon dioxide (CO 2 ) to methane (CH 4 ) is a strategy for sequestering CO 2 . Zero-valent iron (ZVI) has been proposed as an alternative electron donor for the CO 2 reduction to CH 4 . In this study, the effects of ZVI concentrations on the abiotic production of H 2 (without the action of microorganisms) in the first part and on the biological conversion of CO 2 to CH 4 using ZVI as a direct electron donor in the second part were examined. In the abiotic H 2 production, the increase in the ZVI concentration from 16 to 32, 64, and 96 g/L was found to have positive effects on both the amounts of H 2 generated and the rates of H 2 production because the extent of ZVI oxidation positively correlates with increasing surface area. Nevertheless, the increase in ZVI concentration from 96 to 224 g/L did not benefit the H 2 production because the ZVI dissolution was suppressed by the increasing aqueous pH above 10. In the bioconversion of CO 2 to CH 4 using ZVI as an electron donor, the main methanogenesis pathway occurred via hydrogenotrophic methanogenesis at pH 8.7-9.5 driven by the genus Methanobacterium of the class Methanobacteria. At ZVI concentrations of 64 g/L and above, the production of volatile fatty acid (VFA) became clear. Acetate was the main VFA, indicating the induction of homoacetogenesis at ZVI concentrations of 64 g/L and above. In addition, the presence of propionate as the second major VFA suggests the production of propionate from CO 2 and acetate under conditions with high H 2 partial pressure. The results indicated that the pathway for ZVI/CO 2 conversion to CH 4 was competitive between hydrogenotrophic methanogenesis and homoacetogenesis., 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. Published by Elsevier Inc.)

The influence of adding surfactants on the performance of high-solid anaerobic digestion of horticultural waste was extensively investigated in batch systems. Adding Tween series and polyethylene glycol series non-ionic surfactants had positive effects on biogas production, resulting in 370.1 mL/g VS and 256.6 mL/g VS with Tween 60 and polyethylene glycol 300 at a surfactant-to-grass mass ratio of 0.20, while the biogas production of anaerobic digestion without surfactants was 107.54 mL/g VS. The optimal and economically feasible choice was adding Tween 20 at a ratio of 0.08 g/g grass in high-solid anaerobic digestion. A kinetics model reliably represented the relationship between surfactant concentration and biogas production. The mechanism of surfactants working on lignocellulose was investigated. The improvement in high-solid anaerobic digestion by adding surfactants was attributed to the interaction between lignocelluloses and surfactants and the extraction of biodegradable fractions from the porous structure. An economic analysis showed that adding Tween 20 was likely to make a profit and be more feasible than adding Tween 60 and polyethylene glycol 300. This study confirms the enhancement in biogas production from horticultural waste by adding non-ionic surfactants.

This study characterized the microbial community present in the bench scale horizontal-flow anaerobic immobilized biomass bioreactor (HAIB) used in the removal of limonene, a compound present in citrus processing industries. The HAIB was filled with three support materials (coal, polyurethane foam and gravel) which were inoculated with anaerobic sludge. The limonene initial concentration on the substrate ranged from 10 mg/L to 500 mg/L. The analysis of 16S rRNA showed the presence of 22 OTUs (based on ⩾97% sequence identity), distributed in 57 genera, considering three different matrices. Higher relative abundance of phyla was observed as Synergistetes (43-57%), Proteobacteria (32-42%), Firmicutes (7-8%) and Acidobacteria (2-3%). Actinobacteria, Bacterioidetes and Chloroflexi had the lowest relative abundances between 1 and 2%. Synergistaceae family was the predominated group (47.6%-mineral coal, 55.9%-foam and 43.5%-gravel) followed by Syntrophaceae (2.4%-coal, 1.5%-foam and 2.2%-gravel), which kept a syntrophic relationship with methanogenesis (hydrogenotrophic methanogens) to maintain the anaerobic digestion. Among the Proteobacteria phylum, the Pseudomonadaceae family was predominant in the system with 12.0% on coal, 13.1% on foam, and 20.4% on gravel. The metabolic versatility of Pseudomonas sp. makes them an important bioremediation agent by being capable of metabolizing xenobiotic and chemical toxic compounds, thus having great prominence for the limonene removal in the HAIB bioreactor., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Veillonella spp. are nitrate-reducing bacteria with anaerobic respiratory activity that reduce nitrate to nitrite. They are obligate anaerobic, Gram-negative cocci that ferment lactate as the main carbon source and produce short-chain fatty acids (SCFAs). Commensal Veillonella reside in the human body site where lactate level is, however, limited for Veillonella growth. In this study, nitrate was shown to promote the anaerobic growth of Veillonella in the lactate-deficient media. We aimed to investigate the underlying mechanisms and the metabolism involved in nitrate respiration. Nitrate (15 mM) was demonstrated to promote Veillonella dispar growth and viability in the tryptone-yeast extract medium containing 0.5 mM L-lactate. Metabolite and transcriptomic analyses revealed nitrate enabled V. dispar to actively utilize glutamate and aspartate from the medium and secrete tryptophan. Glutamate or aspartate was further supplemented to a medium to investigate individual catabolism during nitrate respiration. Notably, nitrate was demonstrated to elevate SCFA production in the glutamate-supplemented medium, and further increase tryptophan production in the aspartate-supplemented medium. We proposed that the increased consumption of glutamate provided reducing power for nitrate respiration and aspartate served as a substrate for fumarate formation. Both glutamate and aspartate were incorporated into the central metabolic pathways via reverse tricarboxylic acid cycle and were linked with the increased production of acetate, propionate, and tryptophan. This study provides further understanding of the promoted growth and metabolic mechanisms by commensal V. dispar utilizing nitrate and specific amino acids to adapt to the lactate-deficient environment.IMPORTANCENitrate is a pivotal ecological factor influencing microbial community and metabolism. Dietary nitrate provides health benefits including anti-diabetic and anti-hypertensive effects via microbial-derived metabolites such as nitrite. Unraveling the impacts of nitrate on the growth and metabolism of human commensal bacteria is imperative to comprehend the intricate roles of nitrate in regulating microbial metabolism, community, and human health. Veillonella are lactate-utilizing, nitrate-reducing bacteria that are frequently found in the human body site where lactate levels are low and nitrate is at millimolar levels. Here, we comprehensively described the metabolic strategies employed by V. dispar to thrive in the lactate-deficient environment using nitrate respiration and catabolism of specific amino acids. The elevated production of SCFAs and tryptophan from amino acids during nitrate respiration of V. dispar further suggested the potential roles of nitrate and Veillonella in the promotion of human health., Competing Interests: The authors declare no conflict of interest.

We developed a nondestructive three-dimensional microbial visualization method utilizing synchrotron radiation X-ray microscale computed tomography to better understand the relationship between microorganisms and their surrounding habitats. The method was tested and optimized using a mixture of axenic Escherichia coli and Comamonas testosteroni . The osmium-thiocarbohydrazide-osmium method was used to stain all the microbial cells, and gold in situ hybridization was used to detect specific phylogenetic microbial groups. The stained samples were embedded in epoxy resin for microtomographic analysis. Differences in X-ray absorbances were calculated by subtracting the pre-L 3 -edge images from the post-L 3 -edge images to visualize the osmium and gold signals. Although we successfully detected cells stained with osmium, those labeled with gold were not detected, probably because of the insufficient density of gold atoms in the microbial cells. We then applied the developed technique to anaerobic granules and visualized the distribution of microbial cells and extracellular polymeric substances. Empty spaces were highlighted to determine the cavity distribution in granules. Numerous independent cavities of different sizes were identified in the granules. The developed method can be applied to various environmental samples for deeper insights into microbial life in their habitats., Importance: Microorganisms inhabit diverse environments and often form biofilms. One factor that affects their community structure is the surrounding physical environment. The arrangement of residential space within the formed biofilm plays a crucial role in the supply and transportation of substances, as well as the discharge of metabolites. Conventional approaches, such as scanning electron microscopy and confocal laser scanning microscopy combined with fluorescence in situ hybridization, have limitations as they provide information primarily from the biofilm surface and cross-sections. In this study, we developed a method for detecting microorganisms in biofilms using synchrotron radiation X-ray microscale computer tomography. The developed method allows nondestructive three-dimensional observation of biofilms at a single-cell resolution (voxel size of approximately 200 nm), facilitating an understanding of the relationship between microorganisms and their physical habitats., Competing Interests: The authors declare no conflict of interest.

To investigate the effects of microplastics (MPs) on hydrolysis, acidification and microbial characteristics during waste activated sludge (WAS) anaerobic fermentation process, five different kinds of MPs were added into the WAS fermentation system and results indicated that, compared to the control group, the addition of polyvinyl chloride (PVC)-MPs exhibited the least inhibition on volatile fatty acids (VFAs), reducing them by 13.49 %. Conversely, polyethylene (PE)-MPs resulted in the greatest inhibition, with a reduction of 29.57 %. MPs, while accelerated the dissolution of WAS that evidenced by an increase of lactate dehydrogenase (LDH) release, concurrently inhibited the activities of relevant hydrolytic enzymes (α-Glucosidase, protease). For microbial mechanisms, MPs addition affected the proliferation of key microorganisms (norank&#95;f&#95;Bacteroidetes&#95;vadinHA17, Ottowia, and Propioniclava) and reduced the abundance of genes associated with hydrolysis and acidification (pfkb, gpmI, ilvE, and aces). Additionally, MPs decreased the levels of key hydrolytic and acidogenic enzymes to inhibit hydrolysis and acidification processes. This research provides a basis for understanding and unveils impact mechanisms of the impact of MPs on sludge anaerobic fermentation., 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 B.V. All rights reserved.)

Background: Carbon monoxide (CO), hypothetically linked to prebiotic biosynthesis and possibly the origin of the life, emerges as a substantive growth substrate for numerous microorganisms. In anoxic environments, the coupling of CO oxidation with hydrogen (H 2 ) production is an essential source of electrons, which can subsequently be utilized by hydrogenotrophic bacteria (e.g., organohalide-respring bacteria). While Dehalococcoides strains assume pivotal roles in the natural turnover of halogenated organics and the bioremediation of chlorinated ethenes, relying on external H 2 as their electron donor and acetate as their carbon source, the synergistic dynamics within the anaerobic microbiome have received comparatively less scrutiny. This study delves into the intriguing prospect of CO serving as both the exclusive carbon source and electron donor, thereby supporting the reductive dechlorination of trichloroethene (TCE)., Results: The metabolic pathway involved anaerobic CO oxidation, specifically the Wood-Ljungdahl pathway, which produced H 2 and acetate as primary metabolic products. In an intricate microbial interplay, these H 2 and acetate were subsequently utilized by Dehalococcoides, facilitating the dechlorination of TCE. Notably, Acetobacterium emerged as one of the pivotal collaborators for Dehalococcoides, furnishing not only a crucial carbon source essential for its growth and proliferation but also providing a defense against CO inhibition., Conclusions: This research expands our understanding of CO's versatility as a microbial energy and carbon source and unveils the intricate syntrophic dynamics underlying reductive dechlorination., (© 2024. The Author(s).)

Anaerobic ammonium oxidation (anammox), an energy-efficient deamination biotechnology, faces operational challenges in low-temperature environments. Enhancing the metabolic activity of anammox bacteria (AnAOB) is pivotal for advancing its application in mainstream municipal wastewater treatment. Inspired by the metabolic adaptability of AnAOB and based on our previous findings, this work investigated the enhancement of intracellular ATP and NADH synthesis through the exogenous supply of reduced humic acid (HA red ) and H 2 O 2 redox couple, aiming to augment AnAOB activity under low-temperature conditions. Our experimental setup involved continuous dosing of 0.0067 μmol g -1 volatile suspended solid of H 2 O 2 and 10 mg g -1 volatile suspended solid of HA red into a mainstream anammox reactor operated at 15 °C with an influent TN content of 60 mg/L. The results showed that HA red / H 2 O 2 couple succeeded in maintaining the effluent TN at 10.72 ± 0.91 mg l -1 . The specific anammox activity, ATP and NADH synthesis levels of sludge increased by 1.34, 2.33 and 6.50 folds, respectively, over the control setup devoid of the redox couple. High-throughput sequencing analysis revealed that the relative abundance of Candidatus Kuenenia after adding HA red / H 2 O 2 couple reached 3.65 % at the end of operation, which was 5.14 folds higher than that of the control group. Further metabolomics analysis underscored an activation in the metabolism of amino acids, nucleotides, and phospholipids, which collectively enhanced the availability of ATP and NADH for the respiratory processes. These findings may provide guidance on strategy development for improving the electron transfer efficiency of AnAOB and underscore the potential of using redox couples to promote the mainstream application of anammox technology., 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.)

Minimization of antibiotic resistance genes (ARGs) and potential pathogenic antibiotic-resistant bacteria (PARB) during anaerobic digestion (AD) is significantly impacted by temperature. However, knowledge on how ARGs and PARB respond to temperature transition from thermophilic to mesophilic is limited. Here, we combined metagenomic-based with culture-based approaches and revealed the risks of antimicrobial resistance and pathogenicity during transition from 55 °C to 35 °C for AD, with strategies of sharp (ST, one-step by 20 °C/d) and mild (MT, step-wise by 1 °C/d). Results indicated a lower decrease in methane production with MT (by 38.9%) than ST (by 88.8%). Phenotypic assays characterized a significant propagation of multi-resistant lactose-fermenting Enterobacteriaceae and indicator pathogens after both transitions, especially via ST. Further genomic evidence indicated a significant increase of ARGs (29.4-fold), virulence factor genes (1.8-fold) and PARB (65.3-fold) after ST, while slight enrichment via MT. Bacterial succession and enhanced horizontal transfer mediated by mobile genetic elements promoted ARG propagation in AD during transition, which was synchronously exacerbated through horizontal transfer mechanisms mediated by cellular physiological responses (oxidative stress, membrane permeability, bacterial conjugation and transformation) and co-selection mechanisms of biomethanation metabolic functions (acidogenesis and acetogenesis). This study reveals temperature-dependent resistome and pathogenicity development in AD, facilitating microbial risk control., 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 B.V. All rights reserved.)

Nanofiltration (NF) is a promising technology in the treatment of microelectronic wastewater. However, the treatment of concentrate derived from NF system remains a substantial technical challenge, impeding the achievement of the zero liquid discharge (ZLD) goal in microelectronic wastewater industries. Herein, a ZLD system, coupling a two-stage NF technology with anaerobic biotechnology was proposed for the treatment of tetramethylammonium hydroxide (TMAH)-contained microelectronic wastewater. The two-stage NF system exhibited favorable efficacy in the removal of conductivity (96 %), total organic carbon (TOC, 90 %), and TMAH (96 %) from microelectronic wastewater. The membrane fouling of this system was dominated by organic fouling, with the second stage NF membrane experiencing a more serious fouling compared to the first stage membrane. The anaerobic biotechnology achieved a near-complete removal of TMAH and an 80 % reduction in TOC for the first stage NF concentrate. Methyloversatilis was the key genus involved in the anaerobic treatment of the microelectronic wastewater concentrate. Specific genes, including dmd-tmd, mtbA, mttB and mttC were identified as significant players in mediating the dehydrogenase and methyl transfer pathways during the process of TMAH biodegradation. This study highlights the potential of anaerobic biodegradation to achieve ZLD in the treatment of TMAH-contained microelectronic wastewater by NF 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 B.V. All rights reserved.)

Various pretreatments are commonly adopted to facilitate dissolved organic matter (DOM) release from waste activated sludge (WAS) for high-valued volatile fatty acids (VFAs) promotion, while the interplay impact of DOM dynamics transformation on microbial population and metabolic function traits is poorly understood. This work constructed "DOM-microorganisms-metabolism-VFAs" symbiotic ecologic networks to disclose how DOM dynamics variation intricately interacts with bacterial community networks, assembly processes, and microbial traits during WAS fermentation. The distribution of DOM was altered by different pretreatments, triggering the release of easily biodegradable compounds (O/C ratio > 0.3) and protein-like substance. This alteration greatly improved the substrates biodegradability (higher biological index) and upregulated microbial metabolism capacity (e.g., hydrolysis and fatty acid synthesis). In turn, microbial activity modifications augment substance metabolism level and expedite the conversion of highly reactive compounds (proteins-like DOM) to VFAs, leading to 1.6-4.2 fold rise in VFAs generation. Strong correlations were found between proteins-like DOM and topological properties of DOM-bacteria associations, suggesting that high DOM availability leads to more intricate ecological networks. A change in the way communities assemble, shifting from stronger uniform selection in pH10 and USp reactors to increased randomness in heat reactor, was linked to DOM composition alterations. The ecologic networks further revealed metabolic synergy between hydrolytic-acidogenic bacteria (e.g., Bacteroidota and Firmicutes) and biodegradable DOM (e.g., proteins and amino sugars) leading to higher VFAs generation. This study provides a deeper knowledge of the inherent connections between DOM and microbial traits for efficient VFAs biosynthesis during WAS anaerobic fermentation, offering valuable insights for effective WAS pretreatment strategies., 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.)

Nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) is an important methane (CH 4 ) consumption and nitrogen (N) removal pathway in estuarine and coastal wetlands. Antibiotic contamination is known to affect microbially mediated processes; however, its influences on n-DAMO and the underlying molecular mechanisms remain poorly understood. In the present study, using 13 CH 4 tracer method combined with molecular techniques, we investigated the responses of n-DAMO microbial abundance, activity, and the associated microbial community composition to sulfamethazine (SMT, a sulfonamide antibiotic, with exposure concentrations of 0.05, 0.5, 5, 20, 50, and 100 µg L -1 ). Results showed that the effect of SMT exposure on n-DAMO activity was dose-dependent. Exposure to SMT at concentrations of up to 5 µg L -1 inhibited the potential n-DAMO rates (the average rates of nitrite- and nitrate-DAMO decreased by 92.9 % and 79.2 % relative to the control, respectively). In contrast, n-DAMO rates tended to be promoted by SMT when its concentration increased to 20-100 µg L -1 (the average rates of nitrite- and nitrate-DAMO increased by 724.1 % and 630.1 % relative to the low-doses, respectively). Notably, low-doses of SMT suppressed nitrite-DAMO to a greater extent than nitrate-DAMO, indicating that nitrite-DAMO was more sensitive to SMT than nitrate-DAMO. Molecular analyses suggest that the increased n-DAMO activity under high-doses SMT exposure may be driven by changes in microbial communities, especially because of the promotion of methanogens that provide more CH 4 to n-DAMO microbes. Moreover, the abundances of n-DAMO microbes at high SMT exposure (20 and 50 µg L -1 ) were significantly higher than that at low SMT exposure (0.05-5 µg L -1 ). These results advance our understanding of the ecological effects of SMT on carbon (C) and N interactions in estuarine and coastal wetlands., Competing Interests: Declaration of Competing Interest The authors declare no known conflicts of competing financial interest or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The addition of composite conductive materials is being increasingly recognized as a promising strategy to enhance anaerobic digestion (AD) performance. However, the influence of these materials on protein hydrolysis has been poorly documented. Here, a novel magnetic biochar derived from oil sludge and straw was synthesized using different iron sources and successfully applied in sludge AD. Experimental results revealed that magnetic biochar modified by Fe 2+ exhibited excellent electron transfer capacity, moderate magnetization, diverse functional groups (e.g. C=O, C-O=O-), and abundant iron distribution. These characteristics significantly enhanced the hydrolysis of tryptophan-like components, leading to increased methane production (144.44 mL gVS -1 vs 79.72 mL gVS -1 in the control test). Molecular docking analysis revealed that the binding of magnetic biochar related Fe 2+ and Fe 3+ , onto sludge proteins via hydrogen bond played a key role in promoting subsequent protein hydrolysis. Additionally, the noteworthy conservation of protein structures from α-helix and β-sheet to random coil, along with the breakdown of the amide I-associated C=O group and amide III-related CN and NH bonds following the addition of magnetic biochar, accelerated the degradation of sludge protein. Observation of variations in protease activity, coenzyme F420, electron transfer system (ETS), and conductivity within the AD systems, particularly the enrichment of Methanospirillum and Methanosaeta archaea, as well as the Petrimonas, Comamonas, and Syntrophomonas bacteria, suggested that magnetic biochar facilitated a conducive environment by improving hydrolysis-acidification and the direct interspecies electron transfer (DIET) process for acetoclastic methanogens. Moreover, metabolic pathways further proved that tryptophan metobalism and acetoclastic methanogenesis were both facilitated by magnetic biochar. This study provides an in-depth understanding of the impact of magnetic biochar on protein hydrolysis in sewage sludge AD., 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.)

The reliable and efficient nitrite production rate (NPR) through nitritation process is the prerequisite for the efficient running of subsequent processes, like the anammox process and the nitrite shunt. However, there has been scant research on stable and productive nitritation process in recent years. In this study, at a stable hydraulic retention time of 12.0 h and with precise and strict DO control, the upper limit of the NPR was initially investigated using a continuous-flow granular sludge reactor. The NPR of 1.69 kg/m 3 /d with a nitrite production efficiency of 81.97% was finally achieved, which set a record until now in similar research. The median sludge particle size of 270.0 μm confirmed the development of clearly defined granular sludge. The genus Nitrosomonas was the major ammonium oxidizing bacteria. In conclusion, this study provides valuable insights for the practical application of the effective nitritation process driving subsequent nitrogen removal 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 © 2024 Elsevier Inc. All rights reserved.)

Anaerobic co-digestion (AcoD) with kitchen waste (KW) is an alternative utilization strategy for algal bloom waste (AW). However, the kinetic characteristic and metabolic pathway during this process need to be explored further. This study conducted a comprehensive kinetic and metagenomic analysis for AcoD of AW and KW. A maximum co-digestion performance index (CPI) of 1.13 was achieved under the 12% AW addition. Co-digestion improved the total volatile fatty acids generation and the organic matter transformation efficiency. Kinetic analysis showed that the Superimposed model fit optimally (R 2 Adj  = 0.9988-0.9995). The improvement of the kinetic process by co-digestion was mainly reflected in the increase of the methane production from slowly biodegradable components. Co-digestion enriched the cellulolytic bacterium Clostridium and the hydrogenotrophic methanogenic archaea Methanobacterium. Furthermore, for metagenome analysis, the abundance of key genes concerned in cellulose and lipid hydrolysis, pyruvate and methane metabolism were both increased in co-digestion process. This study provided a feasible process for the utilization of AW produced seasonally and a deeper understanding of the AcoD synergistic mechanism from kinetic and metagenomic perspectives., 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. Published by Elsevier Inc.)

Nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) process is a promising wastewater treatment technology, but the slow microbial growth rate greatly hinders its practical application. Although high-level nitrogen removal and excellent biomass accumulation have been achieved in n-DAMO granule process, the formation mechanism of n-DAMO granules remains unresolved. To elucidate the role of functional microbes in granulation, this study attempted to cultivate granules dominated by n-DAMO microorganisms and granules coupling n-DAMO with anaerobic ammonium oxidation (Anammox). After long-term operation, dense granules were developed in the two systems where both n-DAMO archaea and n-DAMO bacteria were enriched, whereas granulation did not occur in the other system dominated by n-DAMO bacteria. Extracellular polymeric substances (EPS) measurement indicated the critical role of EPS production in the granulation of n-DAMO process. Metagenomic and metatranscriptomic analyses revealed that n-DAMO archaea and Anammox bacteria were active in EPS biosynthesis, while n-DAMO bacteria were inactive. Consequently, more EPS were produced in the systems containing n-DAMO archaea and Anammox bacteria, leading to the successful development of n-DAMO granules. Furthermore, EPS biosynthesis in n-DAMO systems is potentially regulated by acyl-homoserine lactones and c-di-GMP. These findings not only provide new insights into the mechanism of granule formation in n-DAMO systems, but also hint at potential strategies for management of the granule-based n-DAMO process., 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 Inc. All rights reserved.)

Biofouling is the main challenge in the operation of anaerobic membrane bioreactors (AnMBRs). Biofouling strongly depends on temperature; therefore, we hypothesize that the interactions and viscoelastic properties of soluble microbial products (SMP) and extracellular polymeric substances (EPS) vary with temperature, consequently influencing membrane permeability. This study compares the performance of an AnMBR operated at a similar permeate flux at two temperatures. The transmembrane pressure (TMP) rose rapidly after 5 ± 2 days at 25 °C but only after 18 ± 2 days at 35 °C, although the reactor's biological performance was similar at both temperatures, in terms of the efficiency of dissolved organic carbon removal and biogas composition, which were obtained by changing the hydraulic retention time. Using confocal laser scanning microscopy (CLSM), a higher biofilm amount was detected at 25 °C than at 35 °C, while quartz crystal microbalance with dissipation (QCM-D) showed a more adhesive, but less viscous and elastic EPS layer. In situ optical coherence tomography (OCT) of an ultra-filtration membrane, fed with the mixed liquor suspended solids (MLSS) at the two temperatures, revealed that while a higher rate of TMP increase was obtained at 25 °C, the attachment of biomass from MLSS was markedly less. Increased EPS adhesion to the membrane can accelerate TMP increase during the operation of both the AnMBR and the OCT filtration cell. EPS's reduced viscoelasticity at 25 °C suggests reduced floc integrity and possible increased EPS penetration into the membrane pores. Analysis of the structures of the microbial communities constituting the AnMBR flocs and membrane biofilms reveals temperature's effects on microbial richness, diversity, and abundance, which likely influence the observed EPS properties and consequent AnMBR fouling., Competing Interests: Declaration of competing interest The authors declare there is no competing interest., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)