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Pinene is a plant volatile monoterpenoid which is used in the fragrance, pesticide, and biofuel industries. Although α-pinene has been synthesized in microbial cell factories, the low synthesis efficiency has thus far limited its production. In this study, the cell growth and α-pinene production of the engineered yeast were decoupled by a dynamic regulation strategy, resulting in a 101.1-fold increase in α-pinene production compared to the control. By enhancing the mevalonate pathway and expanding the cytosolic acetyl-CoA pool, α-pinene production was further increased. Overexpression of the transporter Sge1 resulted in a redistribution of global gene transcription, leading to an increased flux of α-pinene synthesis. By optimizing the aeration flow rate in 3-L bioreactors, the α-pinene production reached 1.8 g/L, which is the highest reported α-pinene production in cell factories. Our research provides insights and fundamentals for the efficient synthesis of monoterpenoids in microbial cell factories., 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 © 2025. Published by Elsevier Ltd.)

Microplastics (MPs) are prevalent in wastewater treatment systems, and their behavior is further complicated after undergoing aging processes. This study explored the impact of original and aged polyvinyl chloride (PVC) MPs on wastewater treatment performance and bacterial communities. Results revealed that Fenton-aging treatment induced surface roughening of the MPs and altered their chemical properties. Prolonged exposure to original and aged PVC MPs severely inhibited the removal of chemical oxygen demand and NH 4 + -N, along with lower sludge concentrations. Additionally, PVC MPs increased the production of loosely-bound extracellular polymeric substances (EPS) and decreased protein levels in tightly-bound fractions. The presence of PVC MPs also shifted the bacterial community, reducing nitrogen removal bacteria while enriching EPS-forming bacteria. Furthermore, exposure to PVC MPs led to a decrease in the abundance of key genes involved in nitrogen metabolism. These findings offer insights into the effects of MPs, especially aged variants, on wastewater treatment 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 © 2025 Elsevier Ltd. All rights reserved.)

Dynamic oxygen fluctuations in activated sludge were investigated to enhance valuable biochemical production during wastewater treatment. Batch experiments compared constant aeration with rapid cycling between oxygen-rich and oxygen-poor states. Fluctuating oxygen concentrations (0-2 mg/L) significantly increased production of valuable biochemicals compared to constant oxygen concentration (2 mg/L). Continuous oxygen perturbations increased free amino acids by 35.7 ± 7.6 % and free fatty acids by 76.4 ± 13.0 %, while intermittent perturbations with anoxic periods enhanced free amino acids by 42.4 ± 8.1 % and free fatty acids by 39.3 ± 7.7 %. Fourteen standard amino acids showed significant increases, and most fatty acids had carbon chain lengths between C12-C22. Mechanistically, oxygen perturbations activated FNR and ArcA regulons, resulting in lower relative abundances of TCA cycle enzymes and higher abundances of amino acid and fatty acid biosynthetic enzymes. These findings demonstrate that controlled oxygen fluctuations in wastewater treatment can enhance the biochemical value of activated sludge with minimal process modifications, facilitating resource 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 © 2025 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The improvement of biogas production in anaerobic digestion (AD) by biochar introduction has been demonstrated. However, the distribution of biochar in the digester and its effect on AD have been seldom explored. In this study, the distribution of biochar and their impact on AD were investigated in a 30 L semi-continuously operated bench-scale anaerobic digester. The results demonstrated that the biochar significantly increased biogas yields by 23.38 % under an organic loading rate (OLR) of 3.0 g VS/L·d. The stability of the AD under an OLR of 4.0 g VS/L·d was also improved by biochar introduction. The increased stirring speed of the digester enhanced the spatial distribution uniformity of biochar and increased biogas production by 5.89 %. Reducing the particle size of biochar improved its spatial distribution uniformity but did not significantly increase biogas production, likely due to excessive microbial accumulation on the biochar, which have caused substrate competition. Biochar aided AD by boosting microbial genera of Syntrophomonas, Bacteroidota, Cloacimonadot, and Methanosaeta, accelerating volatile fatty acids consumption, and improving microorganisms' spatial ecological niches. The economic analysis showed that applying residue-based biochar for biogas production presented superior benefits and greater development potential than residue incineration in the food waste AD process. Overall, this study presented a novel and comprehensive understanding of the biochar distribution and impact on food waste AD in digesters., 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 © 2025. Published by Elsevier Ltd.)

There is limited understanding of the granular partial denitrification/anammox (PD/A) microbiota and metabolic hierarchy specific to municipal wastewater treatment, particularly concerning the multi-mechanisms of functional differentiation and granulation tendencies under high-loading shocks. Therefore, this study utilized fragmented mature biofilm as the exclusive inoculum to rapidly establish a granular PD/A system. Following long-term feeding with municipal wastewater, PD/A process reached a total nitrogen removal efficiency of 97.7%, with anammox contributing over 93%. The dominant filamentous bacteria that supported the granular structure underwent significant changes throughout the operational period. Notably, the mature granular PD/A process demonstrated a distinct metabolic preference for recalcitrant, labile, and xenobiotic organics found in municipal wastewater. The biosynthesis of quorum sensing signaling molecules and core cofactors further enhanced the re-development and substrate metabolic adaptations of PD/A granules in real wastewater environments. This research illuminates the micro-ecological succession and metabolic heterogeneity of the granular PD/A process under mainstream loading., 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 © 2025 Elsevier Ltd. All rights reserved.)

Microalgal-bacteria biofilm shows great potential in low-cost greywater treatment. Accurately predicting treated greywater quality is of great significance for water reuse. In this work, machine learning models were developed for simulating and predicting linear alkylbenzene sulfonate (LAS) removal using 152-days collected data from a battled oxygenic microalgal-bacteria biofilm reactor (MBBfR). By using nine variables including influent LAS, hydraulic retention time (HRT), biofilm density and thickness, specific oxygen production and consumption rates, microalgae and bacteria concentrations, and dissolved oxygen (DO), the support vector machine (SVM) model enabled the accurate LAS removal prediction (training set: R 2  = 0.995, (root mean square error, RMSE) = 0.076, (mean absolute error, MAE) = 0.069; testing set: R 2  = 0.961, RMSE = 0.251, MAE = 0.153). SVM can be also successfully applied for MBBfR operation optimization (HRT = 4.28 h, DO = 0.25 mg/L) that achieving accurate prediction of LAS mineralization., 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.)

This study investigated the performance and phase-specific characteristics of mesophilic co-digestion of food waste (FW) with rice straw (RS) at different RS proportions (40 %, 60 %, and 80 %), as well as mono-digestion of RS. The system achieved optimal performance at 40 % RS content, with a methane yield of 383.8 mL/g-VS and cellulose removal efficiency exceeding 75 %. When RS content increased beyond 60 %, process performance declined notably. At 80 % RS ratio, an imbalance between acidogenesis and methanogenesis led to volatile fatty acids (VFAs) accumulation (1500 mg-COD/L), compromising both efficiency and stability. Notably, during the transition to mono-digestion, the system maintained stable operation, characterized by increased abundance of functional enzymes associated with cellulosic substance hydrolysis and acidogenesis. This enhanced enzymatic activity may be attributed to the microbial adaptation induced by previous co-digestion with FW. These findings provide valuable insights for optimizing mesophilic RS digestion through strategic co-digestion approaches., 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 © 2025 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Miltiradiene serves as a pivotal precursor for the synthesis of numerous abietane-type diterpenes with important pharmacological activities. The endogenous mevalonate (MVA) pathway is tightly regulated in Saccharomyces cerevisiae, which limits the availability of precursors for the heterologous production of miltiradiene. In this study, the orthogonal isopentenol utilization pathway (IUP) was constructed and investigated for its adaptability with mitochondria and peroxisomes in S. cerevisiae for the synthesis of miltiradiene. Compartments combinatorial engineering was used to enhance precursor supply and miltiradiene synthesis, thereby elevating the production of miltiradiene to 146.1 mg/L in S. cerevisiae. Furthermore, an artificial multifunctional enzyme, tSmCPS-tSmKSL-PvPT, was constructed by mimicking the natural multifunctional enzyme to enhance the biosynthesis of miltiradiene in S. cerevisiae strain PCM-MT1, which is capable of producing 414.4 mg/L miltiradiene. Finally, the titer of miltiradiene was increased to 1.02 g/L by fed-batch fermentation in a 5 L bioreactor. This study broadens the application of the IUP in S. cerevisiae by integrating compartmentalization and artificial multifunctional enzymes for the synthesis of diterpenes., 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 © 2025 Elsevier Ltd. All rights reserved.)

This study integrates partial denitrification/Anammox (PD/A) with hydroxyapatite (HAP) crystallization in a single reactor, achieving simultaneous nitrogen and phosphorus removal along with phosphorus recovery. By adjusting pH, sludge concentration, low COD/TN ratio, and applying moderate dissolved oxygen stress, the system operated stably and promoted the synergistic growth of HAP and biomass. Results showed a nitrogen removal efficiency (NRE) of 94.13 % and a phosphorus removal efficiency (PRE) of 73.6 %. Metagenomic analysis revealed that under dissolved oxygen stress, The abundance of Candidatus Brocadia increased from 1 % to 26.1 %, significantly boosting anammox activity. indicating enhanced microbial activity. The upregulation of related genes (sdh, suc, hzs) further boosted AnAOB activity. HAP was identified as the main inorganic component of the granule. This process shows strong potential for nitrogen and phosphorus removal with resource recovery in 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 © 2025 Elsevier Ltd. All rights reserved.)

Efficient nitrogen removal after organic capture is challenging through conventional nitrification-denitrification process. Two biofilm-based anoxic/oxic reactors, with a single intermittent zone (R1) or dual intermittent zones (R2), were compared in treating carbon-limited wastewater. Intermittent aeration integrated partial nitrification-anammox (PNA), partial denitrification-anammox (PDA), and denitrification, with anammox-related pathways contributing over 75% nitrogen removal in both reactors. As nitrogen loading rate increased from 0.14 to 0.19 kg-N m -3 day -1 , nitrogen removal efficiency in R1 dropped from 74.3% to 46.0%, while R2 maintained 76.6% removal at low HRT of 6 h. The dual intermittent aeration strategy improved nitrogen removal capacity by enhancing PNA in the first intermittent zone and reducing effluent fluctuation in the second. Anammox bacteria (Candidatus Brocadia, relative abundance: 0.95-2.48%) were enriched across all zones, supporting efficient PNA and PDA. These findings suggested that dual intermittent aeration enhanced anammox in pre-anoxic processes for carbon limited 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 © 2025 Elsevier Ltd. All rights reserved.)

High-solid anaerobic digestion (HSAD) can be used to treat organic waste. However, the operating stability is limited by the hydraulic conditions; therefore, regulation is essential. The flow field contains a large amount of information that cannot be directly regulated. Dimension reduction research in HSAD could simplify hydrodynamic regulation by removing redundant information from the flow field to reduce the dimensionality. However, to date, no relevant studies have been conducted. In this study, dimension reduction research was conducted to simplify the regulation of the hydraulic conditions of HSAD by constructing a mathematical model based on the simulation results. The HSAD flow velocity distribution model was first proposed, which features the mixing efficiency of the reactor. In addition, an HSAD flow control model with a fitting error below 10% was proposed. The controlled hydraulic enhancement strategy for HSAD in this study provides new insights into mixing performance., 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 hybrid bioreactor combining sulfate-reducing ammonium oxidation (Sulfammox) and Anammox offered potential for simultaneous nitrogen and sulfur removal, but the removal efficiency and microbial mechanism remain unclear. This study demonstrated that in the hybrid bioreactor, the ammonium utilization rate (AUR) of Sulfammox increased by 5.42 times. The promotion of NO 2 - on nitrogen and sulfur conversion in Sulfammox could be attributed to: 1) Increasing extracellular polymers substance (EPS) accelerated the stratification of granule sludge; 2) Increasing the relative abundance of Candidatus Brocadia by 29.55 times and Candidatus Anammoxoglobus by 3.17 times; 3) Upregulating the expression of nitrification (amo, hao and nxr) and sulfur metabolism (sat, aprAB dsr and sox) genes, associated with the pathways NH 4 + →NH 2 OH → NO 2 - →NO 3 - and SO 4 2- →S 2- →SO 4 2- . Moreover, Candida Brocadia sapporoensis emerged as a potential specie of Sulfammox, mediating nitrification by hao and nxr, and sulfate reduction by sat and aprAB, thereby enabling electron transfer between nitrogen and sulfur., 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 © 2025. Published by Elsevier Ltd.)

Continuous high-intensity light exposure can inhibit anaerobic ammonium oxidation (anammox) bacteria activity, though the specific impacts on anammox reactor performance remain unclear. This study investigates the effects of long-term light stress on anammox sludge reactors and explores the use of tea polyphenols as an engineering interventions to mitigate photo oxidation damage. The results showed that the nitrogen removal efficiency (NRE) of the reactor rapidly deteriorated to 41.4 % under 10,000 lx light conditions. However, reactors supplemented with 1 mg·L -1 and 5 mg·L -1 tea polyphenols sustained NREs of 75.2 % and 82.5 %, respectively. The addition of tea polyphenols alleviated oxidative stress by scavenging reactive oxygen species such as ·OH and H 2 O 2 , and by enhancing the activities of antioxidant enzymes including total superoxide dismutase and glutathione peroxidase. Candidatus Kuenenia was negatively impacted by light, while unclassified_f__Brocadiaceae thrived under light stress. These findings provide insights for the development of stable nitrogen removal systems under light exposure., 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 © 2025 Elsevier Ltd. All rights reserved.)

Quorum sensing-regulated microbial behaviors often negatively impact wastewater treatment, leading to issues such as biofouling in membrane bioreactors, filamentous bulking, and resistance gene transfer. Quorum quenching, which counteracts quorum sensing, offers a promising strategy to mitigate these problems. This review aims to highlight overlooked perspectives for its application in microbial aggregates during wastewater treatment. First, the review examines the quorum sensing network present in microbial aggregates and the regulatory role of different quorum sensing systems in bacterial function and behavior during wastewater treatment. The discussions cover hierarchical, parallel, and competitive quorum sensing systems to clarify the interactions among these pathways. A precise quorum quenching strategy is proposed to enhance efficiency based on the type of quorum sensing regulation. Additionally, a bridge is established between the physiological characteristics of quorum quenching bacteria and process parameters to achieve process control over bacterial function and behavior during 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.)

Extracellular polymeric substances (EPS) are well-acknowledged to accelerate microalgal biofilm formation, yet specific role of stratified EPS is unknown. Bacterial biofilm stratified EPS could enrich phosphorus, whether microalgal biofilm stratified EPS could also realize phosphorus or nitrogen enrichment remains unclarified. This study investigated microalgae dominant biofilm growth characteristics and nutrients removal via inoculating microalgae and stratified bacterial EPS at various microalgae:bacteria ratios. Soluble-EPS favored biofilm establishment and chlorophyll synthesis, while loosely-bound (LB-EPS) and tightly-bound EPS (TB-EPS) improved phosphorus removal, and optimum microalgae:bacteria cell count ratio was 1:0.5. Under semi-continuous operation, stable and efficient nutrients removal was observed at hydraulic retention time (HRT) of 2 days. Both nitrogen and phosphorus enrichment by TB-EPS over LB-EPS (respectively up to 7.9 and 23.8 times) were innovatively discovered, with enhanced nutrients turnover efficiency at higher HRTs. This study provided direct evidences regarding the role of stratified EPS on microalgal biofilm development and nutrients turnover., 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 © 2025 Elsevier Ltd. All rights reserved.)

Nanobubble water (NBW) or temperature-phased anaerobic digestion assisted by microbial electrolysis cell (MEC-TPAD) can promote sludge hydrolysis and methanogenesis. However, the role of the combined application of NBW and MEC-TPAD in terms of anaerobic performance and related microbial properties remains unclear. This study investigated the impact of Air-NBW on hydrolysis and methanogenesis of dewatered sludge MEC-TPAD. Under different temperatures, NBW increased ammonia nitrogen by 7.8%-13.7% in the hydrolysis phase and ultimate methane yield by 23.3%-41.5%. NBW can significantly promote hydrolysis under mesophilic-mesophilic conditions, while it can promote substantially methanogenesis under thermophilic-thermophilic conditions. Moreover, NBW increased the diversity and richness of microorganisms in hydrolysis. As to bacteria, NBW increased the relative abundance (RA) of Firmicutes but decreased the RA of Proteobacteria. As to archaea, NBW increased the RA of Methanosarcina in hydrolysis but decreased it in methanogenesis. NBW synchronized with MEC-TPAD improved hydrolysis and methanogenesis of the dewatered sludge digestion 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 © 2025 Elsevier Ltd. All rights reserved.)

Antimicrobial resistance (AMR) threatens public health, necessitating urgent efforts to mitigate the global impact of antibiotic resistance genes (ARGs). Anaerobic digestion (AD), known for volatile solid reduction and energy generation, also presents a feasible approach for the removal of ARGs. This review encapsulates the existing understanding of ARGs and antibiotic-resistant bacteria (ARB) during the AD process, highlighting unresolved challenges pertaining to their detection and quantification. The questions raised and discussed include: Do current ARGs detection methods meet qualitative and quantitative requirements? How can we conduct risk assessments of ARGs? What happens to ARGs when they come into co-exposure with other emerging pollutants? How can the application of internal standards bolster the reliability of the AD resistome study? What are the potential future research directions that could enhance ARG elimination? Investigating these subjects will assist in shaping more efficient management strategies that employ AD for effective ARG 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 © 2025 Elsevier Ltd. All rights reserved.)

This study investigated techno-economic analysis of digestate management systems for a full-scale continuous dry anaerobic digestion plant treating organic fraction of municipal solid waste (OFMSW). A three-stage dewatering system was considered as the baseline scenario (Sc-1) whereas nitrogen removal unit (Sc-2), the incorporation of membrane bioreactor with nitrification downstream (Sc-3) and direct thermal drying of the digestate (Sc-4) were evaluated. The capital cost for the dewatering system with a cake dryer was $4.2 million. For Sc-2, NH 4 -N load could be reduced by 60% and Sc-3 would allow for further reduction in COD and NH 4 -N loads with capital cost of $5.2 and $9.1 million, respectively. To achieve an additional reduction in TN load, an investment of $9.8 million was required in Sc-4. Sc-1 had the lowest operation cost, consequently resulting in the highest income of $22.6/ton, whereas Sc-4 had the highest operation cost, resulting in the lowest income of $16.6/ton OFMSW., 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 © 2025 Elsevier Ltd. All rights reserved.)

This study investigates the production of polyhydroxybutyrate (PHB) using the thermophilic bacterium Caldimonas thermodepolymerans in fed-batch fermentation. This research highlights the potential of thermophilic bacteria in biopolymer production due to their ability to operate at high temperatures, which reduces contamination risks and enhances energy efficiency. Optimal fermentation conditions were identified at a temperature of 50 °C, with the strain achieving a maximum specific growth rate (μ max ) of 0.57 h -1 and high biomass concentration of 63.1 g CDW /L. PHB production reached a peak concentration of 31.9 g/L with a productivity of 1.30 g PHB /L/h. The high cell density approach in fed-batch fermentation not only maximizes the productivity and yield of PHB, but also optimizes the production process, making it more suitable for industrial-scale applications. The findings highlight the potential of thermophilic bacteria as a sustainable solution for enhancing PHB production and advancing biodegradable polymer synthesis., 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 © 2025 Elsevier Ltd. All rights reserved.)

Iron-carbon (Fe-C) based biofilters have shown significant advantages in treating mariculture wastewater by facilitating the mixotrophic heterotrophic nitrification-aerobic denitrification (HNAD) process. However, the effects of Fe-C materials and varying carbon-to-nitrogen (C/N) ratios on N removal and C reduction performance remain insufficiently explored. This study demonstrated that the Fe-C biofilter (R-Fe) achieved significantly higher NO 3 - -N removal efficiency (65.1-96.0 %) compared to the control filter (-12.1-76.9 %) across all tested C/N ratios. Furthermore, the N 2 O emission proportion in R-Fe was reduced by 37.4-42.4 % compared to the control. Increasing the influent C/N ratio enhanced N removal efficiency while reducing the proportion of N 2 O emissions. This improvement correlated with enhanced electron transfer activity and an increased abundance of heterotrophic nitrifying-aerobic denitrifying bacteria (HNADB) and heterotrophic denitrifying bacteria (DNB), while the abundance of autotrophic denitrifying bacteria declined. Strong correlations were observed among microbial electron transfer activity, denitrifying microbial communities, Fe transport genes, denitrification-related functional genes, N removal efficiency, and N 2 O emission proportion, highlighting the critical role of electron transfer activity in microbial N 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 © 2025 Elsevier Ltd. All rights reserved.)

This study introduces a technique utilizing natural pyrrhotite powder as a nucleating agent in four sequencing batch reactors (SBRs) for the treatment of dye wastewater. Through analysis of various factors including pH, pyrrhotite surface free energy, sludge zeta potential, and shifts in microbial communities, the mechanism by which pyrrhotite facilitates the formation of aerobic granular sludge (AGS) is elucidated. Over 140 days of continuous operation under neutral conditions, natural pyrrhotite rapidly cultivated AGS under neutral conditions. The structure of the sludge was compact and the settling properties were satisfactory (SVI 30 /SVI 5 close to 1). Reductions in both sludge zeta potential and interfacial free energy of pyrrhotite correlated with increased hydrophobicity of AGS, leading to enhanced sludge aggregation. Changes in pH, sludge interfacial free energy, and zeta potential were found to influence the microbial community composition and diversity within the sludge.This study provides a novel approach for dye 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.)

This work investigates the optimization of medium-chain carboxylate (MCC) production through xylan mixed-culture monofermentation. The pH screening in batch assays showed that the hydrolysis stage and selectivity towards MCC precursors were optimised at pH 6. Subsequently, a continuous stirred tank reactor (CSTR) and a Sequential Batch Reactor (SBR) were operated at different Hydraulic Retention Times (HRT), revealing that the SBR at HRT 2 days yielded the highest caproic acid since lactic acid availability and chain elongation process were balanced. An enriched medium with yeast extract and vitamins favoured the growth of chain elongators, and therefore, the MCC production. Moreover, cross-feeding interaction between bacteria in xylan fermentation was observed, and Pseudoramibacter was present in the highest caproic acid yields. This work highlights the impact of selecting the proper operational window to optimise one-stage MCC production., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Marta Carballa reports financial support was provided by European Regional Development Fund. Juan Iglesias-Riobó, Miguel Mauricio Iglesias and Marta Carballa report financial support was provided by Xunta de Galicia. 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 © 2025 Elsevier Ltd. All rights reserved.)

Anammox coupled partial S 0 -driven autotrophic denitrification (PS 0 AD) technology represents an innovative approach for removing nitrogen from wastewater. The research highlighted the crucial role of biofilm on sulfur particles in the nitrogen removal process. Further analysis revealed that sulfur-oxidizing bacteria (SOB) are primarily distributed in the inner layer of the biofilm, while anammox bacteria (AnAOB) are relatively evenly distributed in inner and outer layers, with Thiobacillus and Candidatus Brocadia being the dominant species, respectively. Except for anammox and PS 0 AD processes, 15 N isotope labeling tests determined that sulfur reshaped nitrogen metabolism pathways, providing solid evidence for the occurrence of sulfammox process. SOB and AnAOB collaborate in nitrogen and sulfur conversion, with SOB-drived PS 0 AD processes reducing nitrate to nitrite for AnAOB to remove ammonia. Conversely, the nitrate produced from anammox process can be reused by SOB. Metagenomic analyses verified that SOB drove the PS 0 AD process through encoding soxBYZ gene, while AnAOB might play an important role in simultaneously driving the anammox and sulfammox processes. These findings underscore the importance of biofilm and clarify the nitrogen-sulfur cycle mechanisms within the coupled system., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Carbon chain elongation offers a promising pathway for converting waste resources into caproate. However, challenges in yield and selectivity have limited its broader application. To address these limitations, anaerobically digested sludge-derived biochar (ADS-B) was incorporated into the carbon chain elongation process. The findings reveal that the addition of 20 g/L ADS-B resulted in the highest net caproate yield (6.5 g/L) and selectivity (61.1%). Further analysis highlighted that ADS-B's superior physicochemical properties enhanced the conversion of butyrate to caproate and facilitated the colonization of key microorganisms, such as Terrisporobacter and Clostridium, essential for caproate production. Additionally, a life cycle assessment indicated that ADS-B addition effectively reduced the environmental impact of caprate production, with additional potential for further mitigation through feedstock substitution. This study provides critical insights into the application of anaerobically digested sludge-derived biochar for enhancing carbon chain elongation, presenting an alternative approach for waste reutilization., 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 © 2025 Elsevier Ltd. All rights reserved.)

Mitigating the release of extracellular antimicrobial resistance genes (exARGs) from wastewater treatment plants (WWTPs) is crucial for preventing the spread of antimicrobial resistance from human domains into the environment. This study aimed to evaluate the applicability of intI1 as a performance indicator for securing the removal of exARGs at WWTPs. We investigated the reduction of exARGs and intI1 in a full-scale WWTP, where identical wastewater was treated using conventional activated sludge (CAS) and membrane bioreactor (MBR) systems. The log reduction values (LRVs) for exARGs were lower than those for intracellular ARGs (iARGs) across all ARG species and treatment systems. LRVs for exARGs were consistently higher in the MBR than in the CAS. The intI1 exhibited lower LRVs compared to most exARGs, ensuring a minimum LRV of exARG in both CAS and MBR systems. Consequently, intI1 is an effective indicator gene for securing the removal of exARGs., 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 © 2025 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Mainstream anammox faces challenges in adapting to non-optimal temperatures and managing greenhouse gas emissions. This study investigates nitrogen removal and N 2 O emissions in attached-growth anammox reactors subjected to rapid temperature shifts (15-55 °C). Temperature reductions to 15-25 °C had minimal impact on the anammox bacterial populations, with nitrogen removal rates of 0.37±0.11 gN/(L⋅d) and 0.88±0.10 gN/(L⋅d) at 15 °C and 25 °C, respectively. In contrast, increasing temperatures to 45-55 °C significantly diminished both anammox biomass and bioactivity. The reactor at 35 °C exhibited the lowest N 2 O emissions (< 1.0 mgN/(L⋅d)), while emissions rose to approximately 5.0 mgN/(L⋅d) at 15 °C and 3.4 mgN/(L⋅d) at 55 °C (during 295-395 d), primarily due to denitrification performed by coexisting ammonia-oxidizing bacteria and denitrifying microbes. This study provides insights into temperature adaptability and N 2 O emission risks, supporting mainstream anammox applications., 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.)

In this study, waste iron scraps (WIS) were exerted to alleviate sulfide inhibition on anammox bacteria and promote anammox nitrogen removal from sulfide-containing wastewater.Short-term batch experiments showed that WIS-addition led to the anammox bacteria activity increasing by 124.8 % at an initial sulfide concentration of 40 mgS/L. During the long-term experiments, the nitrogen removal rate (NRR) reached to 8.76 kg/(m 3 ·d) in the WIS-added reactor, while the maximum NRR was only 3.77 ± 0.31 kg/(m 3 ·d) in the non-WIS reactor. In contrast to anammox bacteria development in the non-WIS reactor, the relative abundance of Candidatus Kuenenia (1.4-3.7 %) declined significantly in the WIS-added reactor, but novel potential anammox bacteria Brocadiaceae&#95;unclassified (60.1 %-78.6 %) were highly enriched. Overall, the experimental evidence suggested that WIS-addition not only mitigated the sulfide inhibition on anammox bacteria, but also promoted novel anammox bacteria proliferation. The findings of this work provide a promising solution for wide engineering applications of anammox treating sulfide-containing 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 © 2024. Published by Elsevier Ltd.)

Polyhydroxyalkanoates (PHAs) are biodegradable polyesters poised to replace plastics. Mixed culture (MC)-based three-stage processes are effective for carbon recovery from waste biomass, but the energy-intensive PHA synthesis is negatively affected by ammonia nitrogen, inhibiting PHA yield. This study aims to reuse ammonia nitrogen efficiently to mitigate its impact and prevent secondary pollution. PHA production assays under varying MC types, substrate types, feeding modes, and oxygen levels showed that the butyrate type substrate-enriched, high-load, low-oxygen mode (R BC(4)P(1)O(+) ) achieved a PHA conversion ratio of 0.45 g COD/g COD, 1.8 times higher than R BC(2)P(5)O(++) , with reduced energy consumption and CO 2 emissions. Ammonia uptake was 0.06 g NH 3 -N/g PHA at a productivity of 4.54 g/L, showing improved nitrogen recycling. Direct recycling of ammonia nitrogen-containing effluent in the PHA-producing MC enrichment system was performed, and no significant decrease was observed in either the physical properties of the MC flocs or the metrics related to PHA synthesis capacity. These results highlight the feasibility of ammonia reuse and indicate that the soluble microbial products in the effluent have minimal impact on MC enrichment., 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 © 2025 Elsevier Ltd. All rights reserved.)

H 2 -mediated microbial electrosynthesis (MES) could run under a high current density, but the low solubility of H 2 limited its performance. Reducing the H 2 bubble size facilitates H 2 gas-liquid mass transfer and it has been reported to be realized on superaerophobic electrodes. Therefore, we adopted a CoP nanowire-modified nickel foam (CoP-NiF) as the superaerophobic cathode in a H 2 -mediated MES reactor to enhance the methane production from CO 2 . The CoP nanowire modification reduced the average diameter of H 2 bubbles from ∼ 300 μm to ∼ 100 μm, thereby exhibiting a 129 % enhancement of the H 2 mass transfer coefficient (K L a = 0.32 min -1 ). The maximum CH 4 production rate with CoP-NiF cathode exhibited a 27 % improvement (2.31 L/L/d) at a high current density of 166.67 A/m 2 . More importantly, a coulombic efficiency higher than 80 % was achieved in the reactor. These results demonstrated that using superaerophobic cathodes is an efficient way to enhance the performance of H 2 -mediated MES reactors., 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.)

In this study, the effects of fermentation pH and redox potential on the performance of the lactate platform were comprehensively evaluated. The results indicated that the type of acidogenic fermentation was influenced by redox potential, while pH was correlated with volatile fatty acid yield. The highest propionate yield was achieved under anaerobic conditions at a pH of9, with the dominant genus Serpentinicella producing propionate through the acrylate pathway. The highest acetate yield was produced under facultative conditions at a pH of 6. This production was primarily facilitated by the dominant genera unclassified&#95;f&#95;&#95;Enterobacteriaceae and Desulfovibrio, which exhibited significant upregulation of the expression of related genes. Furthermore, ecological processes were employed to establish the relationship between environmental factors and microbial communities. This study emphasized the process of converting lactate into volatile fatty acid, providing a theoretical basis for future strategies aimed at regulating targeted acid 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 Elsevier Ltd. All rights reserved.)

This study aimed to examine the impact of aerobic granular sludge (AGS) sizes on its properties and alginate-like exopolymers (ALE) recovery potential. The AGS was cultivated in a lab-scale bioreactor and categorized into six size classes with 200 μm intervals. There appeared a critical size (400-800 μm) for developing stable AGS structure and excellent ALE recovery. A higher hydrophobicity (74.36 %) and density (1,037 g/L) was observed in AGS 400-600μm than other sizes. Moreover, the highest ALE yield was obtained in ALE 600-800μm (388 mg/g VSS) for its higher abundance of EPS-producers (35.1 %), while the PN content of ALE 400-600μm was higher than other samples. Meanwhile, the concentrations of metal elements within the ALE and AGS identified that there was no bio-accumulation of metal elements in the ALE. This study offers an in-depth understanding of biopolymer recovery from AGS, paving the way for a novel resource recovery strategy through the regulation of AGS sizes., 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.)

A novel approach has been proposed integrating sludge enrichment with intermittent gradient aeration to achieve partial nitrification (PN). Results indicated that this method suppressed nitrite-oxidizing bacteria (NOB) activity while maintained ammonia-oxidizing bacteria (AOB) activity, achieving an 82.87 % nitrite accumulation rate (NAR) during startup. During stable operation, specific ammonia oxidation rate and specific nitrite oxidation rate reached 18.44 and 2.67 mg N/g MLVSS/h, respectively. Shortening anoxic time from 30 to 20 min enhanced ammonia removal efficiency by 12.5 %, increasing NAR to 89.55 %. Further reduction to 5 min yielded 38.46 % and 91.88 %, respectively. AOB abundance enriched from 0.62 % to 22.28 %, with the AOB-to-NOB ratio jumping from 0.22 to 59.58. This research presented a rapid and effective strategy for achieving PN., 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.)

Ammonium sulfate ((NH 4 ) 2 SO 4 ) leaching method to extract rare earth elements (REEs) of mine has produced a large amount of NH 4 + -N-enriched wastewater derived from ore body, leading to many serious environmental pollution problems. This study was the first time to establish an in-situ treatment for real REEs wastewater outside and inside the ore body by an isolated indigenous microorganism. The results stated that Citrobacter sp. X-9 achieved the highest NH 4 + -N removal efficiency among the isolated six microbial strains. Moreover, the microbe to treat the REEs wastewater outside ore body gave the greatest NH 4 + -N removal efficiency under the optimized conditions in the Erlenmeyer flask (250-mL) and bioreactor (10-L). Furthermore, compared to the others' modes, the in-situ treatment by cyclic mode with Citrobacter sp. X-9 possessed superior performance in NH 4 + -N removal efficiency for wastewater inside of ore body, showing that the established in-situ treatment was the potential approach for REEs wastewater purification., 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 production of volatile fatty acids (VFA) through high-solids anaerobic fermentation of organic waste offers a promising route for resource recovery. This study used a batch-mode anaerobic leach bed reactor (LBR) with leachate circulation to ferment the organic fraction of municipal solid waste, producing high concentrations of butyric acid, along with notable amounts of lactic and caproic acids. These results provide valuable insights and underscore the need for process optimization within conventional fermentation systems. To better model the LBR's complex dynamics, the Anaerobic Digestion Model No.1 (ADM1) was modified and extended to account for slow hydrolysis in dry fermentation conditions, thermodynamic constraints imposed by Gibbs Free Energy and hydrogen partial pressures, as well as lactic acid production and chain elongation pathways including homoacetogenesis and caproic acid formation. These enhancements provided deeper insights into high-solids anaerobic fermentation, advancing strategies for improved process control and system optimization., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Adile Evren Tugtas reports financial support was provided by Scientific and Technological Research Council of Turkey. 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.)

A coupled thiosulfate-driven denitrification and anammox (TDDA) process was established to remove nitrogen from wastewater. It was optimized in an up-flow anaerobic sludge blanket reactor using synthetic wastewater, and its reliability was then verified with actual wastewater. The results demonstrated that nitrate, nitrite, and ammonium could be synergistically removed, and the highest total nitrogen removal efficiency reached 97.8% at a loading of 1.39 kgN/(m 3 ·d). Anammox bacteria, primarily Candidatus&#95;Brocadia, were the main contributors to nitrogen removal, while sulfur-oxidizing bacteria such as Thiobacillus and Rhodanobacter played a supportive role. By optimizing substrate conditions to enhance the anammox process, the coupled system attained higher abundances of functional genes such as napA, nirS, hzs, soxXA, and soxYZ, along with the corresponding microbial species. The data suggested that microbial cross-feeding and self-adaptation strategies were key to efficient nitrogen removal by TDDA., 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.)

In order to investigate the mechanisms by which bioaugmentation affects psychrophilic anaerobic digestion (AD), this study introduced a psychrophilic methanogenic culture into the sequencing batch of psychrophilic AD systems. The findings demonstrated that bioaugmentation boosted the abundance of Smithella (23.2 times), Syntrophobacter (9.9 times), and Methanothrix (1.4 times) in the psychrophilic AD systems, accelerating acetate and propionate degradation and improving methane production (26 %). Metagenomic analysis showed that bioaugmentation increased the relative abundance of genes related to propionate degradation and methane production, such as propionyl-CoA synthetase (45 %) and acetyl-CoA synthetase (11 %). At the cellular level, genes related to prevention of cell damage and promotion of membrane fluidity were upregulated. This study revealed the effect of bioaugmentation on microbial metabolic activities related to conversion of propionate to methane and cold tolerance in psychrophilic 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 environmental risk of microplastics (MPs) in aerobic granular sludge (AGS) system is unclear. This study evaluates the effects of non-biodegradable polyvinyl chloride microplastics (PVC-MPs) and biodegradable polylactic acid microplastics (PLA-MPs) on AGS systems. The results showed that both destroyed the performance of AGS systems, with PVC-MPs achieving this by disrupting the AGS structure, while PLA-MPs mainly by causing the expansion of filamentous bacteria induced through the stimulation by lactic acid metabolite (R0: 5.52 ± 0.49 μg/L; R PLA 5: 11.67 ± 0.56 μg/L). Moreover, both MPs inhibited nitrogen removal by disrupting partial nitrification and endogenous denitrification and suppressed key microbes such as Candidatus Competibacter and Nitrosomonas. Metabolic pathway analysis and molecular docking have further confirmed the mechanisms by which MPs affect critical metabolic pathways and key enzymes. Consequently, the hazards of biodegradable MPs to the stable operation of sewage treatment plants should also be of concern., 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.)

Solving the plastic crisis requires high recycling quotas and technologies that allow open loop recycling. Here a biological plastic valorization approach consisting of tandem enzymatic hydrolysis and monomer conversion of post-consumer polyethylene terephthalate into value-added products is presented. Hydrolysates obtained from enzymatic degradation of pre-treated post-consumer polyethylene terephthalate bottles in a stirred-tank reactor served as the carbon source for a batch fermentation with an engineered Pseudomonas putida strain to produce 90mg/L of the biopolymer cyanophycin. Through fed-batch operation, the fermentation could be intensified to 1.4 g/L cyanophycin. Additionally, the upcycling of polyethylene terephthalate monomers to the biosurfactants (hydroxyalkanoyloxy)alkanoates and rhamnolipids is presented. These biodegradable products hold significant potential for applications in areas such as detergents, building blocks for novel polymers, and tissue engineering. In summary, the presented bio-valorization process underscores that addressing challenges like the plastic crisis requires an interdisciplinary approach., 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 Authors. Published by Elsevier Ltd.. All rights reserved.)

In this study, an elemental sulfur (S 0 ) autotrophic denitrification reactor (SADR) and a wood chunk and S 0 mixotrophic denitrification reactor (WSMDR) were constructed with dual-layers to effectively remove nitrate from water using two-inlets and internal recirculation. The denitrification rates were 66-114 and 70-104 g-N/(m 3 ·d) for the SADR and WSMDR, respectively. Sulfate production was 5.5-5.9 and 3.2-4.5 mg SO 4 2- /mg reduced N in the SADR and WSMDR, respectively, being lower than theoretical value. In addition, there was no sulfide emission from either reactor. Chlorobium, Chlorobaculum, Ignavibacterium, Sulfuritalea, and Thiobacillus were involved in nitrate reduction in both reactors. Chlorobiaceae had the highest abundance and played an essential role in maintaining the integrity of the co-occurrence pattern. The abundance of functional genes positively correlated with the denitrification performance. This study demonstrates that the operation of two-inlets and internal recirculation can effectively reduce byproduct generation, thereby promoting the practical application of the SADR and WSMDR., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Qiaochong He reports was provided by National Natural Science Foundation of China. Qiaochong He reports was provided by Zhengzhou Science and Technology Bureau. Qiaochong He reports was provided by Henan University of Technology. 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.)

This paper thoroughly examines how policy incentives impact the economic and environmental sustainability of anaerobic digestion (AD) systems. It uses techno-economic and life cycle analyses, along with real industry data, to explore the entire AD process-from feedstock acceptance to digestate disposal. It evaluates the effects of various U.S. policy crediting programs on the economic viability of different AD pathways for treating sewage sludge and food waste. Tipping fees are identified as the primary driver of profitability, while policy credits play a crucial role in enhancing economic feasibility, particularly for renewable natural gas production. However, future regulatory changes could reshape this economic landscape. All AD pathways are found to significantly reduce greenhouse gas emissions, though economic outcomes are highly sensitive to digestate disposal costs and feedstock tipping fees. Co-digestion with food waste is proposed as a strategy to reduce dependence on policy credits and improve long-term economic 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. Published by Elsevier Ltd.)

Fluctuating pH conditions can affect heavy metal mobility, thereby limiting the efficiency of biofiltration systems (BS). To address this, we developed an innovative rhizosphere sponge, biochar-based bioreactor (RBB), designed to stabilize Cd 2+ removal across a pH range of 5 to 9. RBB consistently outperformed the control, achieving a notable 91.3 % Cd 2+ removal at pH 5. By creating optimized oxygen and redox zoning, the rhizosphere sponge enhanced both biochar surface reactions and microbial activity. Under acidic conditions, biochar facilitated Fe 2+ /Mn 2+ precipitation into stable (oxy)hydroxides, a process further driven by microbial oxidation. Consequently, RBB accumulated 1.54 times more Fe-Mn oxide-bound Cd than the control, effectively reducing Cd 2+ mobility. Additionally, loosely bound extracellular polymeric substances claimed preferential Cd 2+ sequestration after acidification. The stabilized microecology and increased ecological niches, allowing RBB to better buffer against pH fluctuations, presenting it as a robust solution for sustainable heavy metal remediation in variable environments., 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.)

This study investigates a novel approach to mitigate biofouling in membrane bioreactors (MBRs) using a combinational quorum quenching (QQ) strategy. Rhodococcus sp. BH4 and Acinetobacter sp. DKY-1 were employed to disrupt intraspecies N-acyl-homoserine lactones (AHL) and interspecies autoinducer-2 (AI-2) quorum sensing, respectively. BH4 and DKY-1 were immobilized independently and the antibiofouling effects of single QQ beads and 1: 1 mixed QQ beads, both with the same final doses, were compared. While both bead types exhibited high QQ activity, the mixed QQ beads more effectively inhibited microbial biofilm formation, delaying biofouling by two times compared to 1.5 and 1.7 times for the single species beads. Additionally, the mixed QQ MBR demonstrated significantly lower extracellular polymeric substances and a notable reduction in the genus Nitrospira. This combined QQ strategy presents a promising method for enhancing antibiofouling performance in MBRs through targeted disruption of microbial communication., 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.)

Syngas from the gasification of organic wastes represents a promising feedstock for fostering a sustainable bioeconomy. However, its potential is currently constrained by the low-value products generated. Osmolytes, such as hydroxyectoine, are high-value compounds, however, their biological production as isolated osmolytes is not yet cost-effective. This study utilized shotgun genomics and laboratory validation to find a carboxydotrophic, halotolerant bacterium, Hydrogenibacillus schlegelii, that could produce hydroxyectoine using H 2 , CO and CO 2 as the sole source of energy and carbon. Subsequently, NaCl concentration, temperature and syngas composition were optimized in semi-continuous bioreactors. Optimal conversion of CO into hydroxyectoine occurred at a gas composition of 70 %:10 % CO:H 2 (v/v) (44.8 ± 10.1 mg hydroxyectoine ·g biomass -1 ). NaCl concentrations of 5 % significantly enhanced hydroxyectoine content (46.7 ± 9.5 mg hydroxyectoine ·g biomass -1 ), but negatively affected gas consumption. This study opens new perspectives for the valorisation of syngas into hydroxyectoine, and for new cell platforms for pharmaceutical production based on syngas., 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.)

This study evaluated long-term performance of graphene oxide (GO) and magnetic GO (MGO) nanosheets in semi-continuous-flow anaerobic digestion (AD) of municipal sludge over 230 days. At organic loading rates (OLRs) of 2.6 and 3.4 g chemical oxygen demand (COD) fed /L/day, 20 and 200 mg/L of GO and MGO did not affect AD performance. However, at an OLR of 5.2 g COD fed /L/day, where control digesters failed, 20 and 200 mg/L of GO and MGO sustained biogas yields at 190 mL/g COD fed /day, similar to yields at lower OLRs (2.6 and 3.4 g COD fed /L/day). This performance persisted after thedaily nanosheet replenishment stopped. Improvements were due to enhanced conductivity and microbial syntropy. The results showed a strong correlation with previous biochemical methane potential (BMP) assays, positioning BMP as a predictive tool for continuous-flow AD performance. Overall, this study demonstrated potential of GO/MGO nanosheets to improve the stability and efficiency of AD systems under stress., 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 Authors. Published by Elsevier Ltd.. All rights reserved.)

Nitrogen removal via anammox is efficient but challenged by their slow growth. Adding granular activated carbon (GAC) increased the total nitrogen removal rate to 66.99 g-N/m 3 /day, compared to 50.00 g-N/m 3 /day in non-GAC reactor. Both reactors dominated by Candidatus Brocadia (non-GAC: 36.25 %, GAC: 35.5 %) but GAC improved specific anammox activity. Functional metabolic profiling from metagenomic analysis unveiled that GAC enhanced pathways associated with electron shuttle production, potentially promoting intra/extracellular electron transfer. In nitrogen metabolism, GAC is indicated to facilitate anammox N 2 H 4 synthesis process, and inhibit nitrification and full denitrification processes, functioned by Nitrosomonas and Castellaniella which are more abundant in the non-GAC reactor. GAC also enhanced dissimilatory nitrate reduction to ammonium and partial denitrification processes, providing anammox with NH 4 + /NO, which was conducted by Anaerolineae members (29.7 % in GAC-reactor and 7.8 % in non-GAC reactor sludge). This research illuminated the intricate microbial nitrogen cycling networks affected by GAC in anammox 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 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

This study aims at addressing inconsistencies in literature regarding the organic loading rate (OLR) and hydraulic retention time (HRT) effect on caproic acid production, using biomass activity as an indicator. Xylose was fully consumed in the reference CSTR and SBR (HRT = 1-day; OLR = 12 gCOD/(L·d)), but different caproic acid yields (0.02 vs 0.11 Cmol/Cmol-s) were observed, which was linked to differences in biomass activity (12 vs 3.5 g/(g VSS·d)). At HRT 0.5 days, xylose conversion was incomplete and lactic acid and ethanol appeared, reducing caproic acid production. However, increasing xylose concentration in the feeding to 24 g COD/L did not change the caproic acid yield (0.12 Cmol/Cmol-s), which was explained by similar biomass activity as in the reference SBR (4.8 g/(g VSS·d)). These findings indicate that the SBR is the optimal configuration, since it allows maintaining a low biomass activity and therefore a high caproic acid yield., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Marta Carballa reports financial support was provided by European Regional Development Fund. Juan Iglesias-Riobó, Miguel Mauricio Iglesias and Marta Carballa report financial support was provided by Xunta de Galicia. 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 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Two-phase anaerobic digestion (AD) performance is significantly influenced by operating parameters such as temperature and solids retention time (SRT), while their impact on antibiotic resistance genes (ARGs) during the acidogenic (AP) and methanogenic (MP) phases remains unclear. This study assessed the abundance of eight ARGs in full-scale two-phase AD, then operated lab-scale two-phase AD systems to evaluate temperature combinations (thermophilic-thermophilic, thermophilic-mesophilic, mesophilic-thermophilic, and mesophilic-mesophilic) at a constant SRT (AP = 2/MP = 13d) and to further assess different SRTs (AP = 2/MP = 13d and AP = 4/MP = 11d). qPCR results revealed that full-scale two-phase AD reduced total ARGs abundance by 87.70 ± 0.50 %. In lab-scale tests, the thermophilic-thermophilic configuration achieved nearly complete ARGs removal (97.61 ± 0.21 %), while the combination of AP = 4/MP = 11d had the highest removal efficiency (83.39 ± 1.17 %). Network analysis indicated that Firmicutes, Bacteroidota, and Proteobacteria were the primary ARG hosts, with Firmicutes dominant. These findings highlight the optimal operating parameters in two-phase AD for maximizing ARGs removal., 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.)

Biotrickling filter (BTF) technology is inefficient in the treatment of Cl-containing volatile organic compounds (VOCs) such as chlorobenzene (CB). This study adopted non-thermal plasma (NTP) as a pretreatment and conducted in-depth analyses, especially in microorganisms, to investigate strengthening mechanism of a NTP to a BTF in the process. The introduction of NTP enhance efficiency of CB removal from 65 % to 90 %, and CO 2 generation from 60 % to 85 %. It is found that the protein content of the extracellular polymeric substances increases from 212 × 10 -3 mg·g -1 filler to 299 × 10 -3 mg·g -1 filler, thus CB capturing and utilization enhanced. Metagenomic analysis showed that bacteria with CB-degrading properties were enriched in BTF, and CB was involved in cellular metabolism as a carbon source. The presence of active substances from NTP is found to stimulate the ability of BTF treatment. The findings of this study will provide theoretical support for the application of NTP-BTF 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.)

Anaerobic ammonia oxidation (anammox), an energy-efficient technology for treating ammonium-rich wastewater, faces the challenge of antibiotic stress in sewage. This paper systematically evaluated the impact of antibiotics on anammox by considering both inhibitory effects and recovery duration. This review focused on cellular responses, including extracellular polymeric substances (EPS), quorum sensing (QS), and enzymes. Then, the physiological properties of cells and the interactions between nitrogen and carbon metabolism under antibiotic stress were discussed, particularly within the anammoxosome. The microbial community evolution and the development and transfer of antibiotic resistance genes (ARGs) were further analyzed to reveal the resistance mechanisms of anammox. To address the limitations imposed by antibiotics, the development of bio-augmentation and combined processes based on molecular biology techniques, such as bio-electrochemical systems (BES), has been suggested. This review offered new insights into the mechanisms of antibiotic inhibition during the anammox process and aimed to advance their engineering applications., 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.)

A novel biochar/tourmaline polyurethane sponge modified biocarrier (BTP) could enhance Anammox bacteria (AnAOB) enrichment and nitrogen removal performance. With higher hydrophilicity and specific surface area, BTP significantly improved total inorganic nitrogen (TIN) removal efficiency to 80 ± 2 %, compared to unmodified biocarrier of 67 ± 3 % when influent TIN reached 633.9 ± 22.0 mg/L. BTP stimulated the upregulation of amino acid synthases genes abundance and improved protein secretion in extracellular polymer substances (EPS). Moreover, significant increases were found in heme concentration, specific anammox activity and hydrazine dehydrogenase of AnAOB with BTP compared to unmodified biocarrier. Extracellular electron transfer pathway of AnAOB was improved by BTP via upregulating cytochrome C and ferredoxin synthesis. Candidatus Brocadia was the main genus in Anammox biofilm, with relative abundance of 20.1 % and 27.6 % in the control and BTP, respectively, which explained the improvement of nitrogen removal performance., 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.)