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Nitrate photolysis has become an efficient, low-cost and promising technology for emerging contaminants removal, while its performance and mechanism for waste activated sludge (WAS) treatment is still unknown. This study innovatively introduced nitrate photolysis for WAS disintegration, and investigated the effect of nitrate addition (150-375 mg N/L) for short-chain fatty acids (SCFAs) production during anaerobic fermentation (AF). The results showed that nitrate photolysis significantly promoted the SCFAs production from WAS, and peaked at 280.7 mg/g VSS with 7-d fermentation with 150 mg N/L addition (150N-UV), which increased by 8.8-35.0 % and 10.7-23.3 % compared with other photolysis groups and sole nitrate groups. Effective release of the soluble organics was observed in the nitrate photolysis groups during AF, especially soluble proteins, reaching 1505.4 mg COD/L at 9 d in 150N-UV group, promoted by 7.0∼15.7 % than nitrate/nitrate photolysis groups. The model compounds simulation experiment further demonstrated the positive effect of nitrate photolysis on organics hydrolysis and SCFAs accumulation. The result of the radical capture and quenching verified the reactive oxygen species contributed more compared with reactive nitrogen species. Functional group analysis confirmed the effective bioconversion of the macromolecular organics during the fermentation. Moreover, the nitrate photolysis enhanced the enrichment of the functional consortia, including anaerobic fermentation bacteria (AFB), e.g., Fnoticella, Romboutsia, Gracilibacter and Sedimentibacter, and nitrate reducing bacteria (NRB), e.g., Acinerobacter and Ahniella. The macrogenetic analysis further revealed that glycolysis, amino acid metabolism, acetate metabolism and nitrogen metabolism were the dominating metabolic pathways during fermentation, and the abundance of the relevant genes were enhanced in 150N-UV group., 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.)

As the gathering place of urban wastewater, wastewater treatment plants (WWTPs) are indispensable for removing microplastics (MPs), one of the emerging contaminants of great concern, from cities into the natural environment. A reliable and efficient extraction method for MPs, especially in organic-rich matrices, such as sludge samples, is the basis for studying MPs contamination, while it is still lacking. The digestion process, which requires further optimisation, is the most important step during extraction. In this study, we developed and optimised a two-step digestion process to extract MPs and proposed a recommended dosage of digestion reagents based on the mixed liquid volatile suspended solids (MLVSS) level of the sample. Successive addition of 30% H 2 O 2  + 1 M HNO 3 (v:v = 1:1, T = 60 °C, t = 5 h + 5 h) could efficiently extract MPs from sludge samples (over 90%), and the recommended dosage of digestion reagent was 100 ml 30% H 2 O 2 +100 ml 1 M HNO 3 with the sample MLVSS lower than approximately 0.43 g. This new method was also applied to examine the characteristics of MPs in two typical WWTPs (anaerobic-anoxic-oxic and biofilter processes) in Shenzhen. The concentrations of MPs in the influent, effluent and dewatered sludge were approximately 114.00 n/L, 6.00 n/L, and 126.00 n/g (dry weight) in WWTP A, whereas 404.00 n/L, 22.00 n/L, and 204.00 n/g (dry weight) in WWTP B, respectively. Rayon and polyester were the dominant polymers in both the WWTPs. Fibers accounted for the largest proportion of the influent and effluent. Sizes between 0.20-0.50 mm were most detected. This study provides a new and efficient reference method to extract MPs from WWTPs samples, especially sludge sample, with less MPs loss and more beneficial to subsequent identification., 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.)

Biomethane as a sustainable, alternative, and carbon-neutral renewable energy source to fossil fuels is highly needed to alleviate the global energy crisis and climate change. The conventional anaerobic digestion (AD) process for biomethane production from waste(water) streams has been widely employed while struggling with a low production rate, low biogas qualities, and frequent instability. The electric-biologically hybrid microbial electrochemical anaerobic digestion system (MEC-AD) prospects more stable and robust biomethane generation, which facilitates complex organic substrates degradation and mediates functional microbial populations by giving a small input power (commonly voltages < 1.0 V), mainly enhancing the communication between electroactive microorganisms and (electro)methanogens. Despite numerous bioreactor tests and studies that have been conducted, based on the MEC-AD systems, the integrated microbial fingerprints, and cooperation, accelerating substrate degradation, and biomethane production, have not been fully summarized. Herein, we present a comprehensive review of this novel developing biotechnology, beginning with the principles of MEC-AD. First, we examine the fundamentals, configurations, classifications, and influential factors of the whole system's performances (reactor types, applied voltages, temperatures, conductive materials, etc.,). Second, extracellular electron transfer either between diverse microbes or between microbes and electrodes for enhanced biomethane production are analyzed. Third, we further conclude (electro)methanogenesis, and microbial interactions, and construct ecological networks of microbial consortia in MEC-AD. Finally, future development and perspectives on MEC-AD for biomethane production are proposed., 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. Published by Elsevier Ltd.)

To date, the application of marine anammox bacteria (MAB) is still a challenge in saline wastewater treatment due to the low growth rate and high sensitivity. Herein, bioelectrochemical system with applied voltage was exerted for the first time to promote the activity of MAB for removing nitrogen from saline wastewater. At the optimal voltage of 1.5 V, the mean total nitrogen removal rate (TNRR) reached the maximum of 0.65 kg/m 3 •d, which was 27.45% higher than that without applied voltage. Besides, applied voltage reduced the microbial diversity of MAB-based consortia, but the relative abundance of Candidatus Scalindua increased by 4.63% at 1.5 V compared with that without applied voltage. Also, proper applied voltage promoted the secretion of EPS and heme c, which resulted in the enhancement of MAB activity. Based on the remodified Logistic model analysis, the lag time of the nitrogen removal process was shortened by 0.72 h at the voltage of 1.5 V. Furthermore, it was found that higher voltage (> 2.0 V) had a negative effect on the MAB activity for low TNRR of 0.33 kg/m 3 •d (2.5 V). However, TNRR increased back to 0.61 kg/m 3 •d after removing the high applied voltage, which implied that the bioactivity was recoverable after being inhibited. These findings demonstrated that external electrical stimulation is an effective strategy to promote nitrogen removal and MAB activity for treating saline wastewater., (Copyright © 2022. Published by Elsevier Ltd.)

Electrostimulated hydrolysis acidification (eHA) has been used as an efficient biological pre-treatment of refractory industrial wastewater. However, the effects of electrostimulation on the function and assembly of planktonic anaerobic sludge microbial communities are poorly understood. Using 16S rRNA gene and metagenomic sequencing, we investigated planktonic sludge microbial community structure, composition, function, assembly, and microbial interactions in response to electrostimulation. Compared with a conventional hydrolysis acidification (HA) reactor, the planktonic sludge microbial communities selected by electrostimulation promoted biotransformation of the azo dye Alizarin Yellow R. The taxonomic and functional structure and composition were significantly shifted upon electrostimulation with azo dyes degraders (e.g. Acinetobacter and Dechloromonas) and electroactive bacteria (e.g. Pseudomonas) being enriched. More microbial interactions between fermenters and decolorizing and electroactive bacteria, as well as fewer interactions between different fermenters evolved in the eHA microbial communities. Moreover, the decolorizing bacteria were linked to the higher abundance of genes encoding for azo- and nitro-reductases and redox mediator (e.g. ubiquinone) biosynthesis involved in the transformation of azo dye. Microbial community assembly was more driven by deterministic processes upon electrostimulation. This study offers new insights into the effects of electrostimulation on planktonic sludge microbial community function and assembly, and provides a promising strategy for the manipulation of anaerobic sludge microbiomes in HA engineering systems., (Copyright © 2021. Published by Elsevier Ltd.)

Compared with common anaerobic digestion, microbial electrolysis has been proven feasibly to accelerate biodegradation and methanogenesis with the advantages of effective electron flow regulation. However, its actual application and scale-up required a full understanding and further investigation on electrode size and distribution. For making full use of the space of the integrated reactor and improve methane recovery, an effective interior configuration was significant. In this work, three types of reactors with different cathode spatial distributions, that is, different cathode space ratios (ratio of cathode surface area to reaction region volume), were studied to form a good flow pattern for obtaining high methane production. Tracer experiments and numerical simulation were employed simultaneously for understanding the hydrodynamics characters of the interior flow field. The results showed that by increasing the cathode space ratio to 1.33 cm 2 /cm 3 and 2 cm 2 /cm 3 , respectively, better flow patterns with the residence time of 1.336 times and 1.363 times of theoretical hydraulic retention time could be obtained. The stacked structure of nickel meshes was beneficial to prolong the contact time of contaminant and improve the mass transfer. Increasing the cathode space ratio could also enhance the electrochemical performance. Considering the organic removal, methane recovery, electrons generation, and material consumption, the recommended cathode space ratio was 1.33 cm 2 /cm 3 . With this structure, COD removal efficiency reached 93.2 ± 1.9% and 94.1 ± 1.5%, methane production rate reached 332.0 and 334.8 mL CH 4 /L reactor/day, and methane yield was 171.3 and 246.4 mL CH 4 /g COD under the HRT of 24 h and 36 h, respectively., Competing Interests: Declaration of Competing Interest The authors declare no competing interests., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Lately, extracellular electron transfer (EET) is widely disclosed on the surface of the bioelectrodes, and conductive (bio)carriers involved in anaerobic biodegradation/biosynthesis. By electrostimulation, microbial consortia colonize the electrodes and accelerate substrate (waste/wastewater) metabolization on the bioanode or biosynthesize value-added products (methane, acetate, etc.) on the biocathode. However, the connections and contributions of planktonic microbial communities have not been effectually understood. Herein, electromethanogenesis were comprehensively investigated in response to different driving-force modes: intermittent electric field applied by manual on-off or natural solar power and continuous electrical field. Intermittent modes implied preferable electron transfer efficiency, higher methane yield and energy recovery efficiencies from wastewater by the microbes in the bulk solutions. Microbial community analysis revealed that less electroactive microorganisms and acetotrophic methanogens in the bulk solutions were accommodated under the intermittent modes than the continuous electrical field, whereas more fermentative bacteria and hydrogenotrophic methanogens evolved in the intermittent driving modes, implying that the interspecies electron transfer both on and away from the electrodes were favorably regulated. Redundancy and network analysis proved that more complicated ecological interactions were shown in the bulk solutions with the periodic on/off of electrical field. These results hinted that the electrostimulation effectively regulated EET bacteria, even in the bulk solutions, while more efficient electron flow to methane through interspecies electron transfer was developed during the intermittent driving regulation., 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 © 2020 Elsevier Ltd. All rights reserved.)

Free nitrous acid (FNA) has been recently reported to be an effective and eco-friendly inactivator for waste activated sludge (WAS), while the limited decomposition of the extracellular polymeric substance (EPS) matrix hampers resource recovery from WAS. This work employed peracetic acid (PAA) to assist FNA and explored the contribution of co-pretreatment to hydrogen recovery in a prefermentation-bioelectrolysis cascading system. The results showed that co-pretreatment led to approximately 8.8% and 20.4% increases in the exfoliation of particulate proteins and carbohydrates, respectively, from tightly bound EPS (TB-EPS) over that of sole FNA pretreatment. Electron paramagnetic resonance analysis verified that the synergistic effect of FNA, PAA and various generated free radicals was the essential process. This effect further promoted the accumulation of volatile fatty acids (VFAs) after 96 h of prefermentation, and the peak concentration in co-pretreated WAS (AD-FPWAS) was approximately 2.5-fold that in sole FNA-pretreated WAS (AD-FWAS). Subsequently, the cascading utilization of organics in the bioelectrolysis step contributed to efficient hydrogen generation. A total of 10.8 ± 0.3 mg H 2 /g VSS was harvested in microbial electrolysis cells (MECs) fed with AD-FPWAS, while 6.2 ± 0.1 mg H 2 /g VSS was obtained from AD-FWAS. X-ray photoelectron spectroscopy (XPS) revealed the effective decomposition of the phospholipid bilayer in the cytomembrane and the transformation of macromolecular organics into VFAs and hydrogen in the cascading system. Further microbial community analysis demonstrated that co-pretreatment enhanced the accumulation of functional consortia, including anaerobic fermentative bacteria (AFB, 28.1%), e.g., Macellibacteroides (6.3%) and Sedimentibacter (6.9%), and electrochemically active bacteria (EAB, 57.0%), e.g., Geobacter (39.0%) and Pseudomonas (13.6%), in the prefermentation and MEC steps, respectively. The possible synergetic and competitive relationships among AFB, EAB, homo-acetogens, nitrate-reducing bacteria and methanogens were explored by molecular ecological network analysis. From an environmental and economic perspective, this promising FNA and PAA co-pretreatment approach provides new insight for energy recovery from WAS biorefineries., 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 © 2020 Elsevier Ltd. All rights reserved.)

Electroactive microorganisms (EAMs) can act as pseudocapacitor to store energy and discharge electrons on need, while electromethanogens acting as receptor are able to utilize electrons, protons and carbon dioxide for methanization. However, external energy is required to overcome thermodynamical barriers for electromethanogenesis. Herein, electro-driving power by solar light was established to accelerate conversion of waste organics to bioenergy. The intermittent power supply modes were elucidated for favourable performances (e.g., current density, methane production rate, energy recovery efficiencies and economic evaluation), compared with the control driven by continuous applied voltage. It was found that natural intermittent solar-powered mode was more beneficial for microorganisms involved in electron transfer and energy recovery than manual sharp on-off mode. Electrochemistry analysis unrevealed that a higher redox current and lower resistance were exhibited under the solar-powered mode. A high charge storage capacity and electron mobility were found through cytochrome c content and live cells ratio in the solar-power assisted bioreactor. The intermittent power driving modes can regulate electron transfer proteins with capacitive storage behavior in biomass, which helps to understand the responses of functional communities on the stress of intermittent electric field. These findings indicate a promising perspective of microbial biotechnology driven by solar power to boost bioenergy recovery from waste/wastewater., 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 © 2020 Elsevier Ltd. All rights reserved.)

Understanding the fate of overall antibiotic resistance genes (ARGs) during the biological treatment of antibiotic containing wastewater is a central issue for the water ecological safety assessment. Although the microbial electrode-respiration based biotransformation process could significantly detoxify some antibiotic contaminants, e.g. chloramphenicol (CAP), the response of CAP-reducing biocathode microbiome and resistome to continuous electrical stimulation, especially ARGs network interactions, are poorly understood. Here, using highthroughput functional gene array (GeoChip v4.6) and Illumina 16S rRNA gene MiSeq sequencing, the structure, composition, diversity and network interactions of CAP-reducing biocathode microbiome and resistome in response to continuous electrical stimulation were investigated. Our results indicate that the CAP bioelectroreduction process could significantly accelerate the elimination of antibacterial activity of CAP during CAP-containing wastewater treatment compared to the pure bioreduction process. Continuous electrical stimulation could obviously alter both the microbiome and resistome structures and consistently decrease the phylogenetic, functional and overall ARGs diversity and network complexity within the CAP-reducing biofilms. The relative abundances of overall ARGs and specific CAP resistance related major facilitator superfamily (MFS) transporter genes were significantly negatively correlated with the reduction efficiency of CAP to inactive antibacterial product AMCl (partially dechlorinated aromatic amine), which may reduce the ecological risk associated with the evolution of multidrug-resistant bacteria and ARGs during antibiotic-containing wastewater treatment process. This study offers new insights into the response of an antibiotic reducing biocathode resistome to continuous electrical stimulation and provides useful information on the assessment of overall ARGs risk for the bioelectrochemical treatment of antibiotic contaminants., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Cathodic methanogenesis is a promising method for accelerating and stabilising bioenergy recovery in anaerobic processes. The change in composition of microbial (especially methanogenic) communities in response to an applied potential-and especially the associated pH gradient-is critical for achieving this goal, but is not well understood in cathodic biofilms. We found here that the pH-polarised region in the 2 mm surrounding the cathode ranged from 6.9 to 10.1, as determined using a pH microsensor; this substantially affected methane production rate as well as microbial community structure. Miseq sequencing data of a highly conserved region of the mcrA gene revealed a dramatic variation in alpha diversity of methanogens concentrated in electrode biofilms under the applied potential, and confirmed that the dominant microbes at the cathode were hydrogenotrophic methanogens (mostly basophilic Methanobacterium alcaliphilum). These results indicate that regional pH variation in the microenvironment surrounding the electrode is an ecological niche enriched with Methanobacterium., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Methane production was tested in membrane-less microbial electrolysis cells (MECs) under closed-circuit (R CC ) and open-circuit (R OC ) conditions, using glucose as a substrate, to understand the regulatory effects of bioelectrochemistry in anaerobic digestion systems. A dynamic model was built to simulate methane productions and microbial dynamics of functional populations, which were colonized in groups R CC and R OC during the start-up stage. The experiment results showed significantly greater methane production in R CC than R OC , the average methane production of R CC was 0.131 m 3 /m 3 /d, which was 1.4 times higher than that of R OC (0.055 m 3 /m 3 /d). The simulation results revealed that bioelectrochemistry had a significant influence on the abundance of microorganisms involved in acidogenesis and methanogenesis. The abundance of glucose-uptaking microorganisms was 87% of the total biomass in R OC without applied voltage, which was 20% higher than that in R CC (67%) when external voltages were applied between the anode and cathode. The abundance of hydrogenotrophic methanogens in R CC was 6% higher than that in R OC . The simulation results were verified through 16S rDNA high-throughput sequencing analysis. An electron balance analysis revealed that alteration of the acidogenesis type led to more acetate and hydrogen production from glucose fermentation, compared with the situation without bioelectrochemistry. An additional pathway from acetate to hydrogen was introduced by bioelectrolysis. These two factors resulted in significant enhancement of methane production in R CC . Bioelectrolysis process directly contributed to 26% of the total methane production after the start-up stage. When the applied voltages were cut down or decreased, R CC could maintain considerable methane productions, because the microbial communities and electron transfer pathways were already formed. Starting-up with high voltage, but operating under low voltage, could be an energy-favorable strategy for accelerating biogas production in bioelectro-anaerobic bioreactors., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The stress of poised cathode potential condition and carbon source switchover for functional biocathode microbial community influences is poorly understood. Using high-throughput functional gene array (GeoChip v4.2) and Illumina 16S rRNA gene MiSeq sequencing, we investigated the phylogenetic and functional microbial community of the initial inoculum and biocathode for bioelectrochemical reduction of nitrobenzene to less toxic aniline in response to carbon source switchover (from organic glucose to inorganic bicarbonate). Selective transformation of nitrobenzene to aniline maintained in the bicarbonate fed biocathode although nitrobenzene reduction rate and aniline formation rate were significantly decreased compared to those of the glucose-fed biocathode. When the electrical circuit of the glucose-fed biocathode was disconnected, both rates of nitrobenzene reduction and of aniline formation were markedly decreased, confirming the essential role of an applied electric field for the enhancement of nitrobenzene reduction. The stress of poised cathode potential condition led to clear succession of microbial communities from the initial inoculum to biocathode and the carbon source switchover obviously changed the microbial community structure of biocathode. Most of the dominant genera were capable of reducing nitroaromatics to the corresponding aromatic amines regardless of the performance mode. Heterotrophic Enterococcus was dominant in the glucose-fed biocathode while autotrophic Paracoccus and Variovorax were dominant in the bicarbonate-fed biocathode. Relatively higher intensity of diverse multi-heme cytochrome c (putatively involved in electrons transfer) and carbon fixation genes was observed in the biocarbonate-fed biocathode, likely met the requirement of the energy conservation and maintained the nitrobenzene selective reduction capability after carbon source switchover. Extracellular pilin, which are important for biofilm formation and potential conductivity, had a higher gene abundance in the glucose-fed biocathode might explain the enhancement of electro-catalysis activity for nitrobenzene reduction with glucose supply. Dominant nitroaromatics-reducing or electrochemically active bacteria and diverse functional genes related to electrons transfer and nitroaromatics reduction were associated with nitrobenzene reduction efficiency of biocathode communities in response to carbon source switchover., (Copyright © 2014 Elsevier Ltd. All rights reserved.)