Co-occurrence of dominant bacteria and methanogenic archaea and their metabolic traits in a thermophilic anaerobic digester.

Thermophilic anaerobic digestion (TAD) represents a promising biotechnology for both methane energy production and waste stream treatment. However, numerous critical microorganisms and their metabolic characteristics involved in this process remain unidentified due to the limitations of culturable isolates. This study investigated the phylogenetic composition and potential metabolic traits of bacteria and methanogenic archaea in a TAD system using culture-independent metagenomics. Predominant microorganisms identified in the stable phase of TAD included hydrogenotrophic methanogens (Methanothermobacter and Methanosarcina) and hydrogen-producing bacteria (Coprothermobacter, Acetomicrobium, and Defluviitoga). Nine major metagenome-assembled genomes (MAGs) associated with the dominant genera were selected to infer their metabolic potentials. Genes related to thermal resistance were widely found in all nine major MAGs, such as the molecular chaperone genes, Clp protease gene, and RNA polymerase genes, which may contribute to their predominance under thermophilic condition. Thermophilic temperatures may increase the hydrogen partial pressure of Coprothermobacter, Acetomicrobium, and Defluviitoga, subsequently altering the primary methanogenesis pathway from acetoclastic pathway to hydrogenotrophic pathway in the TAD. Consequently, genes encoding the hydrogenotrophic methanogenesis pathway were the most abundant in the recovered archaeal MAGs. The potential interaction between hydrogen-producing bacteria and hydrogenotrophic methanogens may play critical roles in TAD processes.

Bisphenol A affects microbial interactions and metabolic responses in sludge anaerobic digestion.

The widespread use of bisphenol A (BPA) has resulted in the emergence of new pollutants in various environments, particularly concentrated in sewage sludge. This study investigated the effects of BPA on sludge anaerobic digestion, focusing specifically on the interaction of microbial communities and their metabolic responses. While the influence of BPA on methane accumulation is not significant, BPA still enhanced the conversion of soluble COD, protein, and polysaccharides. BPA also positively influenced the hydrolysis-acidogenesis process, leading to 17% higher concentrations of volatile fatty acids (VFAs). Lower BPA levels (0.2-0.5 mg/kg dw) led to decreased hydrolysis and acidogenesis gene abundance, indicating metabolic inhibition; conversely, higher concentrations (1-5 mg/kg dw) increased gene abundance, signifying metabolic enhancement. Diverse methane metabolism was observed and exhibited alterations under BPA exposure. The presence of BPA impacted both the diversity and composition of microbial populations. Bacteroidetes, Proteobacteria, Firmicutes, and Chloroflexi dominated in BPA-treated groups and varied in abundance among different treatments. Changes of specific genera Sedimentibacter, Fervikobacterium, Blvii28, and Coprothermobacter in response to BPA, affecting hydrolysis and acetogenesis. Archaeal diversity declined while the hydrogenotrophic methanogen Methanospirillum thrived under BPA exposure. BPA exposure enabled microorganisms to form structured community interaction networks and boost their metabolic activities during anaerobic digestion. The study also observed the enrichment of BPA biodegradation pathways at high BPA concentrations, which could interact and overlap to ensure efficient BPA degradation. The study provides insights into the digestion performance and interactions of microbial communities to BPA stress and sheds light on the potential effect of BPA during anaerobic digestion.

Metagenomic insights into the toxicity of carbamazepine to functional microorganisms in sludge anaerobic digestion.

Contaminants of emerging concern (CECs) contained in sludge, such as carbamazepine, may be toxic to microorganisms and affect the biogenesis of methane during anaerobic digestion. In this study, different scales of anaerobic digesters were constructed to investigate the inhibitory effect of carbamazepine. Results showed that carbamazepine reduced methane production by 11.3 % and 62.1 % at concentrations of 0.4 and 2 mg/g TS, respectively. Carbamazepine hindered the dissolution of organic matter and the degradation of protein. Carbamazepine inhibited some fermentative bacteria, especially uncultured Aminicenantales, whose abundance decreased by 9.5-93.4 % under carbamazepine stress. It is worth noting that most prior studies investigated the effects of CECs only based on well-known microorganisms, ignoring the metabolisms of uncultured microorganisms. Genome-predicted metabolic potential suggested that 54 uncultured metagenome-assembled genomes (MAGs) associated with acidogenesis or acetogenesis. Therein, uncultured Aminicenantales related MAGs were proved to be acetogenic fermenters, their significant reduction may be an important reason for the decrease of methane production under carbamazepine stress. The toxicity of carbamazepine to microorganisms was mainly related to the overproduction of reactive oxygen species. This study elucidates the inhibition mechanism of carbamazepine and emphasizes the indispensable role of uncultured microorganisms in anaerobic digestion.

Deregulation of Ribosome Biogenesis in Nitrite-Oxidizing Bacteria Leads to Nitrite Accumulation.

Nitrite (NO2-) accumulation caused by nitrite-oxidizing bacteria (NOB) inhibition in nitrification is a double-edged sword, i.e., a disaster in aquatic environments but a hope for innovating nitrogen removal technology in wastewater treatment. However, little information is available regarding the molecular mechanism of NOB inhibition at the cellular level. Herein, we investigate the response of NOB inhibition on NO2- accumulation established by a side-stream free ammonia treatment unit in a nitrifying reactor using integrated metagenomics and metaproteomics. Results showed that compared with the baseline, the relative abundance and activity of NOB in the experimental stage decreased by 91.64 and 68.66%, respectively, directly resulting in a NO2- accumulation rate of 88%. Moreover, RNA polymerase, translation factors, and aa-tRNA ligase were significantly downregulated, indicating that protein synthesis in NOB was interfered during NO2- accumulation. Further investigations showed that ribosomal proteins and GTPases, responsible for bindings between either ribosomal proteins and rRNA or ribosome subunits, were remarkably downregulated. This suggests that ribosome biogenesis was severely disrupted, which might be the key reason for the inhibited protein synthesis. Our findings fill a knowledge gap regarding the underlying mechanisms of NO2- accumulation, which would be beneficial for regulating the accumulation of NO2- in aquatic environments and engineered systems.

Magnetic biochar promotes the risk of mobile genetic elements propagation in sludge anaerobic digestion.

Mobile genetic elements (MGEs) mediated horizontal gene transfer is the primary reason for the propagation of antibiotic resistance genes in environment. The behavior of MGEs under magnetic biochar pressure in sludge anaerobic digestion (AD) is still unknown. This study evaluated the effects of different dosage magnetic biochar on the MGEs in AD reactors. The results showed that the biogas yield was highest (106.68 ± 1.16 mL g-1 VSadded) with adding optimal dosage of magnetic biochar (25 mg g-1 TSadded), due to it increased the microorganism's abundance involved in hydrolysis and methanogenesis. While, the total absolute abundance of MGEs in the reactors with magnetic biochar addition increased by 11.58%-77.37% compared with the blank reactor. When the dosage of magnetic biochar was 12.5 mg g-1 TSadded, the relative abundance of most MGEs was the highest. The enrichment effect on ISCR1 was the most significant, and the enrichment rate reached 158.90-214.16%. Only the intI1 abundance was reduced and the removal rates yield 14.38-40.00%, which was inversely proportional to the dosage of magnetic biochar. Co-occurrence network explored that Proteobacteria (35.64%), Firmicutes (19.80%) and Actinobacteriota (15.84%) were the main potential host of MGEs. Magnetic biochar changed MGEs abundance by affecting the potential MGEs-host community structure and abundance. Redundancy analysis and variation partitioning analysis showed that the combined effect of polysaccharides, protein and sCOD exhibited the greatest contribution (accounted for 34.08%) on MGEs variation. These findings demonstrated that magnetic biochar increases the risk of MGEs proliferation in AD system.

Microplastics influence the fate of antibiotics in freshwater environments: Biofilm formation and its effect on adsorption behavior.

Microplastics (MPs) are substrates available for biofilms colonization in natural water environments. The biofilms formation may enhance the ability of MPs to adsorb harmful contaminants. Herein, we investigated the biofilms formation of three different MPs (PVC, PA and HDPE) in simulated natural environment, and observed the chemical structure, charge property, hydrophobicity and other properties of MPs affect microbial biomass and community composition. More importantly, potential pathogens were found in all three MPs biofilms. Furthermore, the adsorption capacities of original MPs and biological aging MPs for norfloxacin (NOR) was compared. HDPE has the largest adsorption capacity for NOR, while PA has the smallest adsorption capacity for NOR. It was concluded that the formation of biofilms enhanced the adsorption of NOR by 50.60 %, 24.17 % and 46.02 % for PVC, PA and HDPE, respectively. In addition, hydrogen-bond interaction, electrostatic interaction and hydrophobic interaction were found to dominate the adsorption of NOR by MPs. Our study contributed to improve the understanding of the interactions between aging MPs and contaminants in the natural water environments, and provided essential information for ecological risk assessment of MPs.

Advanced MOFs@aerogel composites: Construction and application towards environmental remediation.

Environmental pollution has drawn forth advanced materials and progressive techniques concentrating on sustainable development. Metal-organic frameworks (MOFs) have aroused vast interest resulting from their excellent property in structure and function. Conversely, powdery MOFs in highly crystalline follow with fragility, poor processability and recoverability. Aerogels distinguished by the unique three-dimensional (3D) interconnected pore structures with high porosity and accessible surface area are promising carriers for MOFs. Given these, combining MOFs with aerogels at molecule level to obtain advanced composites is excepted to further enhance their performance with higher practicability. Herein, we focus on the latest studies on the MOFs@aerogel composites. The construction of MOFs@aerogel with different synthetic routes and drying methods are discussed. To explore the connection between structure and performance, pore structure engineering and quantitation of MOFs content are outlined. Furthermore, various types of MOFs@aerogel composites and their carbonized derivatives are reviewed, as well as the applications of MOFs@aerogel for environmental remediation referring to water purification and air clearing. More importantly, outlooks towards these emerging advanced composites have been presented from the perspective of practical application and future development.

Facile synthesis of recyclable 3D gelatin aerogel decorated with MIL-88B(Fe) for activation peroxydisulfate degradation of norfloxacin.

The application of traditional powder catalysts is limited by particle agglomeration and difficult recovery. In this work, a three-dimensional porous aerogel catalyst for organic pollutants degradation in water by activating peroxydisulfate (PDS) was successfully synthesized, which was obtained via directly mixing of MIL-88B(Fe) with sol precursors followed by vacuum freeze-drying and low-temperature calcination. MIL-88B(Fe)/gelatin aerogel-150/PDS (MGA-150/PDS) system displayed satisfactory norfloxacin (NOR) degradation performance, which could remove 98.7% of NOR in 90 min. Its reaction rate constant was 23.2 times higher than the gelatin aerogel/PDS (GA/PDS) system. In addition, Electron paramagnetic resonance (EPR) results and radical trapping experiments revealed both radicals (SO4•-, •OH) and non-radical (1O2) pathways had participated in NOR degradation, of which •OH was dominant. Possible degradation pathways were proposed. Moreover, the high degradation efficiency of NOR by MGA-150 composites could still be reached more than 90.0% even after 10 cycles, and the morphology and chemical structure of MGA-150 composites exhibited no significant changes, indicating the arrestive stability of aerogel composites. This progress not only proposed an effective catalyst for PDS activation, but also expanded views for the design and development of 3D functional materials.

Biofilm on microplastics in aqueous environment: Physicochemical properties and environmental implications.

The excellent properties of plastics make them widely used all over the world. However, when plastics enter the environmental medium, microplastics will inevitably be produced due to physical, chemical and biological factors. Studies have shown that microplastics have been detected in terrestrial, aquatic and atmospheric environments. In addition, the presence of microplastics will provide a new artificial adhesion substrate for biofilms. It has been proved that the formation of biofilms could significantly change some properties of microplastics. Some studies have found that microplastics attached with biofilms have higher environmental risks and eco-toxicity. Therefore, considering the widespread existence of microplastics and the ecological risks of microplastic biofilms, the physical and chemical properties of biofilms on microplastics and their impact on microplastics in aqueous environment are worth reviewing. In this paper, we comprehensively reviewed representative studies in this area. Firstly, this study reviews that the existence of biofilms could change the transport and deposition of microplastics. Subsequently, the presence of biofilms would enhance the ability of microplastics to accumulate pollutant, such as persistent organic pollutants, heavy metals and antibiotics. Moreover, the effect of biofilms on microplastics enrichment of harmful microorganisms is summarized. Finally, some future research needs and strategies are proposed to better understand the problem of biofilms on microplastics.

Addition of nanoparticles increases the abundance of mobile genetic elements and changes microbial community in the sludge anaerobic digestion system.

This study explored the fate of mobile genetic elements (MGEs) in anaerobic digestion (AD) system with four nanoparticles (NPs) added, including carbon NPs, Al2O3 NPs, ZnO NPs, and CuO NPs. 16S rRNA amplicon sequencing and quantitative PCR to investigate the microbial community, MGEs abundance and the potential host in the AD process. The results of high-throughput sequencing showed that ZnO NPs and CuO NPs significantly reduced the microbial diversity and significantly changed the microbial community structure. Simultaneously, the absolute abundance of MGEs increased by 145.01%, 159.67%, 354.70%, and 132.80% on the carbon NPs, Al2O3 NPs, ZnO NPs, and CuO NPs. The enrichment rate of tnpA-03 in ZnO NPs group was the highest, which could reach up to 2854.80%. Co-occurrence analysis revealed that Proteobacteria harbored the vast majority of MGEs followed by Firmicutes. Redundancy analysis and variation partitioning analysis showed that metabolites were the main factors that shifted the succession of bacterial communities. Moreover, there were significant positive correlations between metabolites and part MGEs (such as tnpA-01, tnpA-02, tnpA-03, tnpA-04, tnpA-05, tnpA-07 and ISCR1). This study provides a new perspective that NPs increase the risk of antibiotic resistance through MGEs during AD process.

Co-composting of sewage sludge and Phragmites australis using different insulating strategies.

Insulating strategies are indispensable for laboratory-scale composting reactors, however, current insulation methods interfere with the aerobic fermentation behaviors related to composting. To address this issue, a centre-oriented real-time temperature compensation strategy was designed in this study. Five 9 L reactors (R1-R5) with different insulation strategies were used for the co-composting of dewatered sludge and Phragmites australis and compared. The process performance was assessed by monitoring the temperature, O2 and CO2 emissions, the physical-chemical properties of the composting materials were evaluated by measuring the organic matter (OM), carbon nitrogen ratio (C/N), pH, electrical conductivity (EC), and fluorescence excitation-emission matrix (EEM) spectra. And a 16S rDNA analysis was used to quantify the evolution of bacterial community. The main findings are as follows. Compared with R1 as a control, the insulating strategies can increase the maximum temperature and prolong the thermophilic phase of composting. Comparing R1 and R3 showed that real-time temperature compensation can better restore the real fermentation of the compost. The results showed that R5 had the best composting effect, reaching 69.8 °C, which was 25.1%, 29.7%, 19.3%, and 17.3% higher than R1, R2, R3, and R4, respectively, and remaining in the thermophilic phase for 4.24 d, which is 1.4, 1.5, 1.3, and 0.2 times longer than R1, R2, R3, and R4, respectively. Furthermore, it can significantly reduce the temperature difference between the centre and edge of the reactor, which improved the composting material allocation efficiency and composting process control accuracy, further providing a basis for the actual full-scale composting operation.

Evolutions of antibiotic resistance genes (ARGs), class 1 integron-integrase (intI1) and potential hosts of ARGs during sludge anaerobic digestion with the iron nanoparticles addition.

In this work, we investigated the impact of iron nanoparticle, including magnetite nanoparticles (Fe3O4 NPs) and nanoscale zero-valent iron (nZVI), on the anaerobic digestion (AD) performance. Moreover, the evolutions of antibiotic resistance genes (ARGs), class 1 integrons-integrase (intI1) and potential hosts of ARGs were also investigated. The optimal addition of Fe3O4 NPs and nZVI to promote methane production was 0.5 g/L and 1 g/L, which led to 22.07% and 23.02% increase in methane yield, respectively. The degradation rate of organic matter was also enhanced with the addition of Fe3O4 NPs or nZVI. The results of high-throughput sequencing showed that the reactors with iron NPs exhibited significant differences in microbial community structure, compared to the reactors with the non­iron NPs. Iron NPs have caused the relative abundance of the dominant bacteria (Proteobacteria, Firmicutes and Actinobacteria) generally decreased, while the dominant archaea (Euryarchaeota) increased in AD sludge. Quantitative PCR results revealed that iron NPs accelerated the reductions in total absolute abundance of ARGs, especially a beta-lactamase resistance encoded gene (blaOXA). Network analysis displayed that the attenuation of ARGs was mainly attributed to the decline of potential hosts (Proteobacteria, Firmicutes and Actinobacteria). Meanwhile, environmental factors (such as pH, soluble chemical oxygen demand and heavy metals) were also strongly correlated with ARGs.

Effects of different conductive nanomaterials on anaerobic digestion process and microbial community of sludge.

The effects of four conductive nanomaterials (nano-carbon powder, nano-Al2O3, nano-ZnO, nano-CuO) on sludge anaerobic digestion (AD) performance and microbial community were investigated through a 36-day fermentation experiment. Results showed that biogas production enhanced by 16.9% and 23.4% with nano-carbon powder and nano-Al2O3 added but decreased by 90.2% and 17.3% with nano-ZnO and nano-CuO. Total solids (TS) removal efficiency was increased by 38.73% and 27.11% with nano-carbon powder and nano-Al2O3 added but decreased by 70.67% and 43.70% with nano-ZnO and nano-CuO. Kinetic analysis indicated four conductive nanomaterials could shorten the lag phase of AD sludge with an average rate of 51.75%. 16S rRNA amplicon sequencing results demonstrated microbes such as Syntrophomonas and Methanosaeta were enriched in nano-carbon powder and nano-Al2O3 reactors. However, microbial community diversity and richness were both inhibited by adding nano-ZnO and nano-CuO. Redundancy analysis (RDA) revealed that genera belong to Firmicutes and Chloroflexi could conduce to methanogenesis process.

Influence of nanoscale zero-valent iron and magnetite nanoparticles on anaerobic digestion performance and macrolide, aminoglycoside, ß-lactam resistance genes reduction.

The effect of nanoscale zero-valent iron (NZVI) and magnetite nanoparticles (Fe3O4 NPs) on anaerobic digestion (AD) performance was investigated through a series of 100-day semi-continuous mesophilic anaerobic digestions. The results indicated that biogas production had increased by 24.44% and 21.66% with the addition of 0.5 g/L Fe3O4 NPs and 1.0 g/L NZVI, respectively. Besides, the abundance of five widespread antibiotic resistance genes (ARGs) (ermF, ermA, ermT, aac(6')-IB, blaOXA-1) was also studied. The decrease in abundance of aac(6')-IB and blaOXA-1 was observed during the AD process with an average removal rate of 95.69% and 44.82%, respectively. Most of the ARGs, especially ermA and ermT, were less abundant in NZVI group compared with control group. The overall results suggested that the addition of NZVI and Fe3O4 NPs contributed to a better sludge anaerobic digestion performance, and NZVI was beneficial to the removal of some ARGs.

Enhanced mesophilic anaerobic digestion of waste sludge with the iron nanoparticles addition and kinetic analysis.

As the functional material, iron nanoparticles effectively promote anaerobic digestion (AD) process, including the hydrolysis-acidification process and the biogas production. In this study, nano zero-valent iron (nZVI) and Fe3O4 nanoparticles (Fe3O4 NPs) were added to AD reactors respectively. The AD process was evaluated by the reactors performances, including pH, biogas yields and compositions, as well as the removal ratio of total solids (TS), volatile solids (VS) and soluble chemical oxygen demand (sCOD). Three models (first-order kinetic model, transfer function model and Cone model) were used to explore the kinetics of AD biogas production. The results showed that adding appropriate dose of nZVI or Fe3O4 NPs enhanced anaerobic digestibility of sludge. The highest cumulative biogas yield of 140.34 L with 0.5 g L-1 nZVI and 137.13 L with 1 g L-1 Fe3O4 NPs were obtained by the 80 days of mesophilic operation, respectively. Cumulative biogas productions of these two reactors were significantly enhanced up to 15.70% and 13.44%. TS removal rates reached >70% in all AD reactors with iron nanoparticles, and the highest sCOD removal rates of nZVI and Fe3O4 NPs digesters on the 80th day were 88.22% and 77.63%, respectively. The results of the three-day fermentation experiment and the kinetic parameters showed that the nZVI or Fe3O4 NPs enhanced the hydrolysis-acidification process of the AD, which eventually promoted biogas production. The Cone model was satisfied with the experimental results, which could be used to evaluate the kinetics of AD with iron nanoparticles more reasonably.

Halide Ion-Induced Switching of Gold Nanozyme Activity Based on Au-X Interactions.

The influence of halide ion on the peroxidase activity of protein-modified gold nanoparticles (AuNPs) has been explored, based on the Au-X interaction directed binding of halide ion to AuNPs. Due to different Au-X interactions, halide has been demonstrated to display different switching behaviors to the catalytic activity of protein-modified AuNPs. Presented is the finding that iodide can rapidly inhibit the enzyme activity of CM-AuNP nanozyme effectively. Iodide-mediated irreversible inhibition is due not to I--induced aggregation of AuNP but to the Au-I bond-induced blocking of active sites of AuNP nanozyme. I- switching efficiency was found to be strongly dependent on the surface density of modifiers and the intrinsic property of the modifier. Similar to iodide, bromide can also inhibit the enzyme activity effectively, but its inhibition behavior is reversible. Due to the weak Au-Cl interaction, chloride has no influence on the enzyme activity of CM-AuNP at low ion concentration and exhibits weak activity inhibition at high ion concentration. Fluoride shows no influence on the activity of gold nanozyme due to the absence of Au-F interaction. Our results have improved a profound understanding of anion-mediated AuNP nanozyme activity because of their interfacial interaction and provided guidance in the further utilization of nanozyme in numerous areas.

Adsorption Behavior of Low-Concentration Imidazolium-Based Ionic Liquid Surfactant on Silica Nanoparticles.

The adsorption behavior of imidazolium-based ionic liquid surfactant ([C12mim]Br) on silica nanoparticles (NPs) has been studied with turbidity, isothermal titration microcalorimetry, fluorescence spectroscopy, and dynamic light scattering (DLS) measurements. Both the electrostatic attraction and the hydrogen bonding interaction between silica NP and [C12mim]Br play crucial roles during [C12mim]Br monomers binding to silica NPs at low surfactant concentration, and the hydrophobic effect leads to formation of micelle-like aggregates on silica NP surfaces with the further increase of surfactant concentration. Furthermore, it is found that sodium halide salts favor the adsorption of [C12mim]Br on silica NP surfaces by decreasing the electrostatic repulsions. Anions with more hydrophobicity and the ability to form hydrogen bonding have more pronounced effect. Compared with DTAB, [C12mim]Br has much stronger binding ability with silica NPs at pH 7.0. More interestingly, [C12mim]Br can still form micelle-like aggregates on silica NP surfaces, but DTAB cannot at pH 2.0. The hydrogen bonding between the imidazolium ring and silica NPs is the principal contributor to these observations. Our results will contribute to the elucidation of silica NP/cationic surfactant interaction from molecular scale and the widely applications of silica NP/surfactant systems in practice.

Synthesis, structure-activity relationship analysis and kinetics study of reductive derivatives of flavonoids as Helicobacter pylori urease inhibitors.

In a continuing study for discovering urease inhibitors based on flavonoids, nineteen reductive derivatives of flavonoids were synthesized and evaluated against Helicobacter pylori urease. Analysis of structure-activity relationship disclosed that 4-deoxy analogues are more potent than other reductive products. Out of them, 4',7,8-trihydroxyl-2-isoflavene (13) was found to be the most active with IC50 of 0.85 µM, being over 20-fold more potent than the commercial available urease inhibitor, acetohydroxamic acid (AHA). Kinetics study revealed that 13 is a competitive inhibitor of H. pylori urease with a Ki value of 0.641 µM, which is well matched with the results of molecular docking. Biological evaluation and mechanism study of 13 suggest that it is a good candidate for discovering novel anti-gastritis and anti-gastric ulcer agent.