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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 addition of exogenous materials is a commonly reported method for promoting the anaerobic digestion (AD) of sludge. However, most exogenous materials are nano-sized and their use encounters problems relating to a need for continuous replenishment, uncontrollability and non-recyclability. Here, magnetic porous microspheres (MPMs), which can be controlled by magnetic forces, were prepared and used to enhance the methanogenesis of sludge. It was observed that the MPMs were spherical particles with diameters of approximately 100 µm and had a stable macroporous hybrid structure of magnetic cores and polymeric shells. Furthermore, the MPMs had good magnetic properties and a strong solid-liquid interfacial electron transfer ability, suggesting that MPMs are excellent carriers for methanogenic consortia. Experimental results showed that the addition of MPMs increased methane production and the proportion of methane in biogas from AD by 100.0 % and 21.2 %, respectively, indicating the MPMs notably enhanced the methanogenesis of sludge. Analyses of variations in key enzyme activities and electron transfer in sludge samples with and without MPMs in AD revealed that the MPMs significantly enhanced the activities of key enzymes involved in hydrolysis, acidification and methanation. This was achieved mainly by enhancing the extracellular electron transfer to strengthen the proton motive force on the cell membrane, which provides more energy generation for methanogenic metabolism. A careful examination of the variations in the morphology, pore structure and magnetism of the MPMs before and after AD revealed that the MPMs increased the prevalence of many highly active anaerobes, and that this did not weaken the magnetic performance. The microbial community structure and metatranscriptomic analysis further indicated that the acetotrophic methanogens (i.e., Methanosaeta) were mainly in a free state and that CO 2 -reducing methanogens (i.e., Methanolinea and Methanobacterium) mainly adhered to the MPMs. The above synergistic metabolism led to efficient methanogenesis, which indicates that the MPMs optimised the spatial ecological niche of methanogenic consortia. These findings provide an important reference for the development of magnetic porous materials promoting 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.)

Anaerobic digestion (AD) is a promising technique for sludge treatment and resource recovery. Metals are very important components of sludge and can have substantial effects on its complex nature and microbial activity. However, systematic reviews have not addressed how metals in sludge affect AD and how they can be regulated to improve AD. This paper comprehensively reviews the effects of metals on the AD of sludge from both abiotic and biotic perspectives. First, we introduce the contents and basic characteristics (e.g., chemical forms) of intrinsic metals in sewage sludge. Then, we summarise the main mechanism by which metals influence sludge properties and the methods for removing metals and thus improving AD. Next, we analyze the effects of both intrinsic and exogenous metals on the enzymes and microbial communities involved in anaerobic bioconversion, focusing on the types, critical concentrations and valence states of the metals. Finally, we propose ideas for future research on the roles of metals in the AD of sludge. In summary, this review systematically clarifies the roles of metals in the AD of sludge and provides a reference for improving AD by regulating these metals., 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 © 2023 Elsevier B.V. All rights reserved.)

The recovery of renewable bioenergy from anaerobic digestion (AD) of sludge is a promising method to alleviate the energy problem. Although methane can be effectively recovered through sludge pretreatment by cation exchange resin (CER), the simultaneous enhancement of hydrogen and methane generation from AD using CER has not been extensively investigated. Herein, the effect of CER on the sequential recovery of hydrogen and methane and the corresponding mechanisms were investigated. When CER is introduced, the maximum increases for the hydrogen and methane production are 104.7 % and 35.3 %, respectively, confirming the sequential enhancement effects of CER on the hydrogen and methane production. Analyses of the variations in the main biochemical components with and without the effect of CER demonstrate that CER promotes sludge organic solubilisation, hydrolysis, and acidification in both hydrogen- and methane-production stages. Moreover, investigations of variations in the solid-liquid interfacial thermodynamics and removal rates of main multivalent metals of sludge reveal that the ion exchange reactions between the CER and sludge in the hydrogen-production stage provide the direct driving force of effective contact between bacteria and organic particulates. Additionally, the residual effect of the CER during methane production reduces the energy barrier for mass transfer and provides a driving force for this transfer. Further analyses of the microbial community structure and metagenomics indicate that CER directly drives the enrichment of hydrogen-producing bacteria (+ 15.1 %) and key genes encoding enzymes in the hydrogen-production stage. Moreover, CER indirectly induces the enrichment of methane-producing anaerobes (e.g. Methanosaeta: + 16.7 %, Methanosarcina: + 316.5 %); enhances the bioconversion of different substrates into methyl-coenzyme M; and promotes the metabolism pathway of acetoclastic process and CO 2 reduction in the methane-production stage. This study can provide valuable insights for simultaneously enhancing the production of hydrogen and methane from AD through sequential recovery., 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 © 2023 Elsevier Ltd. All rights reserved.)

As a by-product of industry, waste iron scraps (WIS) are low-cost and widely available, which was potential for the development of iron-assisted anammox. In this study, the feasibility of adding WIS to enhance the nitrogen removal of the anammox process (also called WIS-assisted anammox) was demonstrated. Results indicated that the WIS-assisted anammox reactors performed a 15-35% higher nitrogen removal efficiency than that of the control. Compared to the sludge from the control, the sludge from the WIS-assisted anammox reactors had a higher iron content (78-113 g kg -1 SS) and a better specific anammox activity (10.8-15.5 mg N g -1 VSS h -1 ). The enhanced growth of the anammox bacteria (related to Ca. Kuenenia stuttgartiensis with 99% similarity) in the WIS-assisted anammox reactors was also confirmed by high-throughput sequencing and qPCR. Furthermore, the functional genes predicted by PICRUSt2 revealed a higher level of hydroxylamine oxidoreductase (hao)-like proteins expression of the biomass from the WIS-assisted anammox reactors, implying that the hydroxylamine-related anammox pathway was promoted. Additionally, the observation of cytoplasmic nitrate reductase (narG), copper-containing nitrite reductase (nirK), and nitric oxide reductase (norB) suggested that the introduction of WIS might promote the denitrification ability. This was correlated to the lower ΔNO 3 - /ΔNH 4 + ratio observed in these WIS-assisted anammox reactors. Overall, the WIS-assisted anammox offers a sustainable nitrogen removal process for wastewater treatment with waste iron recycling., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Partial nitritation is necessary for the implementation of the mainstream anammox (anaerobic ammonium oxidation) process in wastewater treatment plants. However, the difficulty in outcompeting nitrite-oxidizing bacteria (NOB) at mainstream conditions hinders the performance of partial nitritation. The present work aimed to develop a high-rate partial nitritation process for low-ammonium wastewater treatment at low temperatures by seeding aerobic granules. Experimental results suggested that both stratified structure of nitrifiers developed in the granules and sufficient residual ammonium concentration (18-35 mg N L -1 ) in the bulk liquid contributed to efficient NOB repression. With the hydraulic retention time progressively shortened from 1.0 to 0.17 h, the influent nitrogen loading rate of the partial nitritation process reached 6.8 ± 0.4 kg N m -3 d -1 even at 10-15 °C. The high concentration (7.5 gVSS L -1 ) and activity (0.48 g N g -1 VSS d -1 at 11 °C) of granular sludge made the reactor possess an overcapacity evaluated by the ratio between the actual ammonium oxidation rate of the granules and their maximum potential. The overcapacity helped the reactor to face the adverse effect of decreasing temperatures. Overall, this work indicated the great potential of applying aerobic granules to achieve high-rate partial nitritation at mainstream conditions. Moreover, anammox bacteria with a relative abundance of 2.8% was also identified in the partial nitritation granules at the end of this study, suggesting that the granules provided a habitable niche for anammox bacteria growth. Note that these results cannot fully relate to the treatment of real domestic/municipal wastewater, they are a source of important information increasing the knowledge about low temperature partial nitrification., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

High-solid sludge anaerobic digestion leads to increased organic matters in digester supernatant, which promotes heterotrophic competition and reduces anaerobic ammonium oxidation (anammox) retention. This research demonstrated effective anammox retention by hydrocycloning during a two-stage deammonification. Anammox retention was evaluated by dividing large (>0.425 mm), medium (0.25 to 0.425 mm), and small (<0.25 mm) aggregate fractions via a sieve, whereby Candidatus Kuenenia abundance in each size aggregate was measured to be 16.8%, 5.0%, and 0.9% respectively. After hydrocyclone separation, large particles took up only 1.7% to 2.7% in the overflow discharge (upper discharge from the reactor), whereas its initial proportion was 19.4%, indicating limited anammox loss. The volume ratio change of large particles to total aggregates was defined for particle breakdown evaluation. Breakdown (23% to 32% large particles) occurred mainly during pumping (influenced by pump frequency and type), while little happened in the hydrocyclone. This research provided methods to use a sieve to evaluate anammox retention by hydrocyclone during high-solid sludge anaerobic digester supernatant deammonification, and information for reducing particle breakdown, pumping selection, and separation optimization., 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 © 2021 Elsevier B.V. All rights reserved.)

[Display omitted] • A single stage PN/A process treating low strength wastewater was started rapidly. • The biofilm growth process on porous polyurethane hydrogel carrier was studied. • Biomass entrapment by interconnected micropores dominated the initial colonization. • AOB was surface-attached growth, anammox colonies proliferated as embedded growth. Media-supported biofilm is a powerful strategy for growth and enrichment of slow-growing microorganisms. In this study, a single-stage nitritation-anammox process treating low-strength wastewater was successfully started to investigate the biofilm development on porous polyurethane hydrogel carrier. Suspended biomass migration into the carrier and being entrapment by its internal interconnected micropores dominated the fast initial colonization stage. Both surface-attached growth and embedded growth of microbes occurred during the following accumulation stage. Fluorescence in situ hybridization analysis of mature biofilm indicated that ammonium-oxidizing bacteria located at the outer layers featured a surface-attached growth, while anammox microcolonies housed in the inner layers proliferated as an embedded-like growth. In this way, the growth rate of anammox bacteria (predominated by Candidatus Kuenenia) could be 0.079 d-1. The anammox potential of the biofilm reactor reached 1.65 ± 0.3 kg/m3/d within two months. This study provides novel insights into nitritation-anammox biofilm formation on the porous polyurethane hydrogel carrier. [ABSTRACT FROM AUTHOR]

Achieving mainstream nitritation with aerobic granules is attractive based on increasing evidence but generally treating artificial low-ammonium wastewater. Real municipal wastewater is much more complex in composition, the behavior of the nitritation granules would be different when treating real municipal wastewater. Herein, the response of nitritation granules to influent shift from artificial low-ammonium (35–40 mg/L) wastewater to anaerobically pre-treated municipal wastewater (MWW pre-treated) was investigated at low temperatures. Results showed that MWW pre-treated caused the outgrowth of filamentous bacteria on the granule surface and developed into finger-like structures, which in turn resulted in the decrease of the overall granular sludge settleability. Batch-tests and microbial analysis indicated the functional and microbial differentiation between the newly formed fluffy exterior and the original compact granule. The fluffy exterior was dominated by genus Flavobacterium (66.6%) and primarily functioned as COD removal, whereas the nitrifiers (mainly Nitrosomonas) were still located in the compact core and performed nitritation. Moreover, the heterotrophs-dominated fluffy exterior hindered the oxygen transfer towards nitrifiers located in the compact granule and thereby facilitated the stable NOB repression in the granule particularly at low temperatures (<10 °C). Finally, gradual recovery of the granular sludge morphology and settleability occurred after the influent reverted to synthetic low-ammonium wastewater. Overall, this work demonstrated that the feeding of MWW pre-treated only caused morphological changes of the nitritation granules, but its structural and functional stability could be maintained stably. [Display omitted] • Granule-based nitritation was maintained treating anaerobically pretreated sewage. • COD removal occurred primarily in external flocs, while nitritation in compact core. • External flocs promoted NOB repression by increasing oxygen transfer resistance. [ABSTRACT FROM AUTHOR]