Radiation and temperature drive diurnal variation of aerobic methane emissions from Scots pine canopy.

Methane emissions from plant foliage may play an important role in the global methane cycle, but their size and the underlying source processes remain poorly understood. Here, we quantify methane fluxes from the shoots of Scots pine trees, a dominant tree species in boreal forests, to identify source processes and environmental drivers, and we evaluate whether these fluxes can be constrained at the ecosystem-level by eddy covariance flux measurements. We show that shoot-level measurements conducted in forest, garden, or greenhouse settings; on mature trees and saplings; manually and with an automated CO2-, temperature-, and water-controlled chamber system; and with multiple methane analyzers all resulted in comparable daytime fluxes (0.144 ± 0.019 to 0.375 ± 0.074 nmol CH4 g-1 foliar d.w. h-1). We further find that these emissions exhibit a pronounced diurnal cycle that closely follows photosynthetically active radiation and is further modulated by temperature. These diurnal patterns indicate that methane production is associated with diurnal cycle of sunlight, indicating that this production is either a byproduct of photosynthesis-associated biochemical reactions (e.g., the methionine cycle) or produced through nonenzymatic photochemical reactions in plant biomass. Moreover, we identified a light-dependent component in stand-level methane fluxes, which showed order-of-magnitude agreement with shoot-level measurements (0.968 ± 0.031 nmol CH4 g-1 h-1) and which provides an upper limit for shoot methane emissions.

Aerobic methane production in Scots pine shoots is independent of drought or photosynthesis.

Shoot-level emissions of aerobically produced methane (CH4) may be an overlooked source of tree-derived CH4, but insufficient understanding of the interactions between their environmental and physiological drivers still prevents the reliable upscaling of canopy CH4 fluxes. We utilised a novel automated chamber system to continuously measure CH4 fluxes from the shoots of Pinus sylvestris (Scots pine) saplings under drought to investigate how canopy CH4 fluxes respond to the drought-induced alterations in their physiological processes and to isolate the shoot-level production of CH4 from soil-derived transport and photosynthesis. We found that aerobic CH4 emissions are not affected by the drought-induced stress, changes in physiological processes, or decrease in photosynthesis. Instead, these emissions vary on short temporal scales with environmental drivers such as temperature, suggesting that they result from abiotic degradation of plant compounds. Our study shows that aerobic CH4 emissions from foliage are distinct from photosynthesis-related processes. Thus, instead of photosynthesis rates, it is more reliable to construct regional and global estimates for the aerobic CH4 emission based on regional differences in foliage biomass and climate, also accounting for short-term variations of weather variables such as air temperature and solar radiation.

The hidden roots of wetland methane emissions.

Wetlands are the largest natural source of methane (CH4 ) globally. Climate and land use change are expected to alter CH4 emissions but current and future wetland CH4 budgets remain uncertain. One important predictor of wetland CH4 flux, plants, play an important role in providing substrates for CH4 -producing microbes, increasing CH4 consumption by oxygenating the rhizosphere, and transporting CH4 from soils to the atmosphere. Yet, there remain various mechanistic knowledge gaps regarding the extent to which plant root systems and their traits influence wetland CH4 emissions. Here, we present a novel conceptual framework of the relationships between a range of root traits and CH4 processes in wetlands. Based on a literature review, we propose four main CH4 -relevant categories of root function: gas transport, carbon substrate provision, physicochemical influences and root system architecture. Within these categories, we discuss how individual root traits influence CH4 production, consumption, and transport (PCT). Our findings reveal knowledge gaps concerning trait functions in physicochemical influences, and the role of mycorrhizae and temporal root dynamics in PCT. We also identify priority research needs such as integrating trait measurements from different root function categories, measuring root-CH4 linkages along environmental gradients, and following standardized root ecology protocols and vocabularies. Thus, our conceptual framework identifies relevant belowground plant traits that will help improve wetland CH4 predictions and reduce uncertainties in current and future wetland CH4 budgets.

Unveiling the Occurrence and Non-Negligible Role of Amino Sugars in Waste Activated Sludge Fermentation by an Enriched Chitin-Degradation Consortium.

Polysaccharides in extracellular polymeric substances (EPS) can form a hybrid matrix network with proteins, impeding waste-activated sludge (WAS) fermentation. Amino sugars, such as N-acetyl-d-glucosamine (GlcNAc) polymers and sialic acid, are the non-negligible components in the EPS of aerobic granules or biofilm. However, the occurrence of amino sugars in WAS and their degradation remains unclear. Thus, amino sugars (∼6.0%) in WAS were revealed, and the genera of Lactococcus and Zoogloea were identified for the first time. Chitin was used as the substrate to enrich a chitin-degrading consortium (CDC). The COD balances for methane production ranged from 83.3 and 95.1%. Chitin was gradually converted to oligosaccharides and GlcNAc after dosing with the extracellular enzyme. After doing enriched CDC in WAS, the final methane production markedly increased to 60.4 ± 0.6 mL, reflecting an increase of ∼62%. Four model substrates of amino sugars (GlcNAc and sialic acid) and polysaccharides (cellulose and dextran) could be used by CDC. Treponema (34.3%) was identified as the core bacterium via excreting chitinases (EC 3.2.1.14) and N-acetyl-glucosaminidases (EC 3.2.1.52), especially the genetic abundance of chitinases in CDC was 2.5 times higher than that of WAS. Thus, this study provides an elegant method for the utilization of amino sugar-enriched organics.

Quorum Sensing Enhances Direct Interspecies Electron Transfer in Anaerobic Methane Production.

Direct interspecies electron transfer (DIET) provides an innovative way to achieve efficient methanogenesis, and this study proposes a new approach to upregulate the DIET pathway by enhancing quorum sensing (QS). Based on long-term reactor performance, QS enhancement achieved more vigorous methanogenesis with 98.7% COD removal efficiency. In the control system, methanogenesis failure occurred at the accumulated acetate of 7420 mg of COD/L and lowered pH of 6.04, and a much lower COD removal of 41.9% was observed. The more significant DIET in QS-enhancing system was supported by higher expression of conductive pili and the c-Cyts cytochrome secretion-related genes, resulting in 12.7- and 10.3-fold improvements. Moreover, QS enhancement also improved the energy production capability, with the increase of F-type and V/A-type ATPase expression by 6.3- and 4.2-fold, and this effect probably provided more energy for nanowires and c-Cyts cytochrome secretion. From the perspective of community structure, QS enhancement increased the abundance of Methanosaeta and Geobacter from 54.3 and 17.6% in the control to 63.0 and 33.8%, respectively. Furthermore, the expression of genes involved in carbon dioxide reduction and alcohol dehydrogenation increased by 0.6- and 7.1-fold, respectively. Taken together, this study indicates the positive effects of QS chemicals to stimulate DIET and advances the understanding of the DIET methanogenesis involved in environments such as anaerobic digesters and sediments.

Mitigation of Triclocarban Inhibition in Microbial Electrolysis Cell-Assisted Anaerobic Digestion.

Triclocarban (TCC), as a widely used antimicrobial agent, is accumulated in waste activated sludge at a high level and inhibits the subsequent anaerobic digestion of sludge. This study, for the first time, investigated the effectiveness of microbial electrolysis cell-assisted anaerobic digestion (MEC-AD) in mitigating the inhibition of TCC to methane production. Experimental results showed that 20 mg/L TCC inhibited sludge disintegration, hydrolysis, acidogenesis, and methanogenesis processes and finally reduced methane production from traditional sludge anaerobic digestion by 19.1%. Molecular docking revealed the potential inactivation of binding of TCC to key enzymes in these processes. However, MEC-AD with 0.6 and 0.8 V external voltages achieved much higher methane production and controlled the TCC inhibition to less than 5.8%. TCC in the MEC-AD systems was adsorbed by humic substances and degraded to dichlorocarbanilide, leading to a certain detoxification effect. Methanogenic activities were increased in MEC-AD systems, accompanied by complete VFA consumption. Moreover, the applied voltage promoted cell apoptosis and sludge disintegration to release biodegradable organics. Metagenomic analysis revealed that the applied voltage increased the resistance of electrode biofilms to TCC by enriching functional microorganisms (syntrophic VFA-oxidizing and electroactive bacteria and hydrogenotrophic methanogens), acidification and methanogenesis pathways, multidrug efflux pumps, and SOS response.

Optimizing alkali-pretreatment dosage for waste-activated sludge disintegration and enhanced biogas production yield.

The rapid transition towards modernization and industrialization led to an increase in urban population, resulting in paramount challenge to municipal sewage sludge management. Anaerobic digestion (AD) serves as a promising venue for energy recovery from waste-activated sludge (WAS). Addressing the challenge of breaking down floc structures and microbial cells is crucial for releasing extracellular polymeric substances and cytoplasmic macromolecules to facilitate hydrolysis and fermentation process. The present study aims to introduce a combined process of alkaline/acid pre-treatments and AD to enhance sludge digestion and biogas production. The study investigates the influence of alkali pretreatment at ambient temperature using four alkali reagents (NaOH, Ca(OH)2, Mg(OH)2, and KOH). The primary goal is to provide insights into the intricate interplay of alkali dosages (0.04-0.12 g/gTS) on key physic-chemical parameters crucial for optimizing the pre-treatment dosage. Under the optimized alkaline/acid pre-treatment condition, the TSS reduction of 18%-30% was achieved. An increase in sCOD concentration (24%-50%) signifies the enhanced hydrolysis and solubilization rate of organic substrate in WAS. Finally, the biomethane potential test (BMPT) was performed for pre-treated WAS samples. The maximum methane (CH4) yield was observed in combination A1 (244 mL/g) and D1 (253 mL/g), demonstrating the pivotal role of alkali optimization in enhancing AD efficiency. This study serves as a valuable resource to policymakers, researchers, and technocrats in addressing challenges associated to sludge management.

Biochar alleviates inhibition effects of humic acid on anaerobic digestion: Insights to performances and mechanisms.

To recover methane from waste activated sludge through anaerobic digestion (AD) is one promising alternative to achieve carbon neutrality for wastewater treatment plants. However, humic acids (HAs) are one of the major compositions in waste activated sludge, and their accumulation performs inhibition effects on AD. This study investigated the potentials of biochar (BC) in alleviating inhibition effects of HAs on AD. Results showed that although the accumulated HAs reduced methane yield by 9.37% compared to control, the highest methane yield, 132.6 mL CH4/g VSS, was obtained after adding BC, which was 45.9% higher than that in HA group. Mechanism analysis showed that BC promoted the activities of hydrolase such as protease and α-glucosidase, which were 69.7% and 29.7% higher than those in HA group, respectively. The conversion of short-chain fatty acids was accelerated. In addition, the evolutions of electroactive microorganisms like Clostridium_sensu_stricto_13 and Methanosaeta were consistent with the activitiies of electron transfer and the contents of cytochrome c. Furthermore, parts of HAs rather than all of them were adsorbed by BC, and the remaining free HAs and BC formed synergistic effects on methanogenesis, then both CO2 reduction and acetoclastic methanogenesis pathways were improved. The findings may provide some solutions to alleviate inhibition effects of HAs on AD.

The interplay of soil physicochemical properties, methanogenic diversity, and abundance governs methane production potential in paddy soil subjected to multi-decadal straw incorporation.

Straw incorporation holds significant promise for enhancing soil fertility and mitigating air pollution stemming from straw burning. However, this practice concurrently elevates the production and emission of methane (CH4) from paddy ecosystems. Despite its environmental impact, the precise mechanisms behind the heightened CH4 production resulting from long-term straw incorporation remain elusive. In a 32-year field experiment featuring three fertilization treatments (CFS-chemical fertilizer with wheat straw, CF-chemical fertilizer, and CK-unamended), we investigated the impact of abiotic (soil physicochemical properties) and biotic (methanogenic abundance, diversity, and community composition) factors on CH4 production in paddy fields. Results revealed a significantly higher CH4 production potential under CFS treatment compared to CF and CK treatments. The partial least squares path model revealed that soil physicochemical properties (path coefficient = 0.61), methanogenic diversity (path coefficient = -0.43), and methanogenic abundance (path coefficient = 0.29) collectively determined CH4 production potential, explaining 77% of the variance. Enhanced soil organic carbon content and water content, resulting from straw incorporation, emerged as pivotal factors positively correlated with CH4 production potential. Under CFS treatment, lower Shannon index of methanogens, compared to CF and CK treatments, was attributed to increased Methanosarcina. Notably, the Shannon index and relative abundance of Methanosarcina exhibited negative and positive correlations with CH4 production potential, respectively. Methanogenic abundance, bolstered by straw incorporation, significantly amplified overall potential. This comprehensive analysis underscores the joint influence of abiotic and biotic factors in regulating CH4 production potential during multi-decadal straw incorporation.

Enhancement of anaerobic digestion from food waste via inert substances based on metagenomic analysis: Oxidative phosphorylation and metabolism.

The application of anaerobic digestion (AD) in the treatment of food waste (FW) has become widespread. However, the presence of inert substances, such as bones, ceramics, and shells, within FW introduces a degree of uncertainty into the AD process. To clarify this intricate issue, this study conducted an in-depth investigation into the influence of inert substances on AD. The results revealed that when inert substances were present at a concentration of 0.08 g/g VSS, methane productivity in the AD process was significantly augmented by 86%. Subsequent investigations suggested that this positive effect was primarily evident in various biochemical processes, including solubilization, hydrolysis acidification, methanogenesis, and the accumulation of extracellular polymeric substances. Metagenomic analysis showed that inert substances enhance the relative abundance of hydrolytic bacteria and have a pronounced impact on the relative abundance of hydrogenotrophic methanogens (Methanosarcina) and acetotrophic methanogens (Methanobacterium). Additionally, inert substances significantly increased the relative abundance of functional genes in oxidative phosphorylation, a pivotal pathway for ATP synthesis. Furthermore, inert substances had a substantial effect on the functional genes related to the metabolic pathways associated with methanogenesis (both hydrogenotrophic and acetotrophic). This comprehensive study shed light on the substantial impact of inert substances on the AD of food waste, contributing to an enhanced understanding of the underlying mechanisms of anaerobic fermentation.

Heterogeneous g-C3N4/polyaniline composites enhanced the conversion of organics into methane during anaerobic wastewater treatment.

In this study, g-C3N4/PANI was prepared by in situ oxidative polymerization. Graphite-phase carbon nitride (g-C3N4) with surface defects was deposited onto the surface of conductive polyaniline (PANI) to form a p-n heterojunction. This construction aimed to create an efficient heterogeneous catalyst, increasing the surface defect level and active sites of the composite, and augmenting its capability to capture and transfer extracellular electrons under anaerobic conditions. This addresses the challenge of low efficiency in direct interspecies electron transfer between bacteria and archaea during anaerobic digestion for methane production. The results showed that the prepared g-C3N4/PANI increased the CH4 yield and CH4 production rate by 82% and 96%, respectively. Notably, the conductivity and XPS test results showed that the ratio of g-C3N4 to PANI was 0.15, and the composite exhibited favorable conductivity, with a uniform distribution of pyrrolic nitrogen, pyridinic nitrogen, and graphitic nitrogen, each accounting for approximately 30%. Furthermore, g-C3N4/PANI effectively enhanced the metabolic efficiency of intermediate products such as acetate and butyrate. Analysis of the microbial community structure revealed that g-C3N4/PANI led to a significant increase in the abundance of hydrogenotrophic methanogen Methanolinea (from 48% to 64%) and enriched Clostridium (a rise of 1%) with direct interspecies electron transfer capability. Microbial community function analysis demonstrated that the addition of g-C3N4/PANI boosted the activities of key enzymes involved in anaerobic digestion, including phosphate transacetylase (PTA), phospho-butyryl transferase (PTB), and NAD-independent lactate dehydrogenase (NNLD), by 47%, 135%, and 153%, respectively. This acceleration in enzymatic activity promoted the metabolism of acetyl-CoA, butyryl-CoA, and pyruvate. Additionally, the function of ABC transporters was enhanced, thereby improving the efficiency of material and energy exchange among microorganisms.

Effect of a blend of cinnamaldehyde, eugenol, and Capsicum oleoresin on methane emission and lactation performance of Holstein-Friesian dairy cows.

This study aimed to investigate the effect of administering a standardized blend of cinnamaldehyde, eugenol, and Capsicum oleoresin (CEC) to lactating dairy cattle for 84 d (i.e., 12 wk) on enteric CH4 emission, feed intake, milk yield and composition, and body weight. The experiment involved 56 Holstein-Friesian dairy cows (145 ± 31.1 d in milk at the start of the trial; mean ± standard deviation) in a randomized complete block design. Cows were blocked in pairs according to parity, lactation stage, and current milk yield, and randomly allocated to 1 of the 2 dietary treatments: a diet including 54.5 mg of CEC/kg of DM or a control diet without CEC. Diets were provided as partial mixed rations in feed bins, which automatically recorded individual feed intake. Additional concentrate was fed in the GreenFeed system that was used to measure emissions of CO2, CH4, and H2. Feeding CEC decreased CH4 yield (g/kg DMI) by on average 3.4% over the complete 12-wk period and by on average 3.9% from 6 wk after the start of supplementation onward. Feeding CEC simultaneously increased feed intake and body weight, and tended to increase milk protein content, whereas no negative responses were observed. These results must be further investigated and confirmed in longer-term in vivo experiments.

Lactation modeling and the effects of rotational crossbreeding on milk production traits and milk-spectra-predicted enteric methane emissions.

Rotational crossbreeding has not been widely studied in relation to the enteric methane emissions of dairy cows, nor has the variation in emissions during lactation been modeled. Milk infrared spectra could be used to predict proxies of methane emissions in dairy cows. Therefore, the objective of this work was to study the effects of crossbreeding on the predicted infrared proxies of methane emissions and the variation in the latter during lactation. Milk samples were taken once from 1,059 cows reared in 2 herds, and infrared spectra of the milk were used to predict milk fat (mean ± SD; 3.79 ± 0.81%) and protein (3.68 ± 0.36%) concentrations, yield (21.4 ± 1.5 g/kg dry matter intake), methane intensity (14.2 ± 2.0 g/kg corrected milk), and daily methane production (358 ± 108 g/d). Of these cows, 620 were obtained from a 3-breed (Holstein, Montbéliarde, and Viking Red) rotational mating system, and the rest were purebred Holsteins. Milk production data and methane traits were analyzed using a nonlinear model that included the fixed effects of herd, genetic group, and parity, and the 4 parameters (a, b, c, and k) of a lactation curve modeled using the Wilmink function. Milk infrared spectra were found to be useful for direct prediction of qualitative proxies, such as methane yield and intensity, but not quantitative traits, such as daily methane production, which appears to be better estimated (450 ± 125 g/d) by multiplying a measured daily milk yield by infrared-predicted methane intensity. Lactation modeling of methane traits showed daily methane production to have a zenith curve, similar to that of milk yield but with a delayed peak (53 vs. 37 d in milk), whereas methane intensity is characterized by an upward curve that increases rapidly during the first third of lactation and then slowly till the end of lactation (10.5 g/kg at 1 d in milk to 15.2 g/kg at 300 d in milk). However, lactation modeling was not useful in explaining methane yield, which is almost constant during lactation. Lastly, the methane yield and intensity of cows from 3-breed rotational crossbreeding are not greater, and their methane production is lower than that of purebred Holsteins (452 vs. 477 g/d). Given the greater longevity of crossbred cows, and their lower replacement rate, rotational crossbreeding could be a way of mitigating the environmental impact of milk production.

Estimates of genetic parameters for rumination time, feed efficiency, and methane production traits in first-lactation Holstein cows.

The large-scale recording of traits such as feed efficiency (FE) and methane emissions (ME) for use in genetic improvement programs is complex, costly, and time-consuming. Therefore, heritable traits that can be continuously recorded in dairy herds and are correlated with FE and ME traits could provide useful information for genetic evaluation. Rumination time has been suggested to be associated with FE, methane production (MeP; ME in g/d), and production traits at the phenotypic level. Therefore, the objective of this study was to investigate the genetic relationships among rumination time (RT), FE, methane and production traits using 7,358 records from 656 first-lactation Holstein cows. The estimated heritabilities were moderate for RT (0.45 ± 0.14), MeP (0.36 ± 0.12), milk yield (0.40 ± 0.08), fat yield (0.29 ± 0.06), protein yield (0.32 ± 0.07), and energy-corrected milk (0.28 ± 0.07), but were low and nonsignificant for FE (0.15 ± 0.07), which was defined as the residual of the multiple linear regression of DMI on energy-corrected milk and metabolic body weight. A favorable negative genetic correlation was estimated between RT and MeP (-0.53 ± 0.24), whereas a positive favorable correlation was estimated between RT and energy-corrected milk (0.49 ± 0.11). The estimated genetic correlation of RT with FE (-0.01 ± 0.17) was not significantly different from zero but showed a trend of a low correlation with dry matter intake (0.21 ± 0.13). These results indicate that RT is genetically associated with MeP and milk production traits, but high standard errors indicate that further analyses should be conducted to verify these findings when more data for RT, MeP, and FE become available.

Long-term effects of 3-nitrooxypropanol on methane emission and milk production characteristics in Holstein-Friesian dairy cows.

The objective was to determine the long-term effect of 3-nitrooxypropanol (3-NOP) on CH4 emission and milk production characteristics from dairy cows receiving 3-NOP in their diet for a full year, covering all lactation stages of the dairy cows. Sixty-four late-lactation Holstein-Friesian cows (34% primiparous) were blocked in pairs, based on expected calving date, parity, and daily milk yield. The experiment started with an adaptation period of 1 wk followed by a covariate period of 3 wk in which all cows received the same basal diet and baseline measurements were performed. Directly after, cows within a block were randomly allocated to 1 of 2 dietary treatments: a diet containing on average 69.8 mg 3-NOP/kg DM (total ration level, corrected for intake of nonsupplemented GreenFeed bait) and a diet containing a placebo. Forage composition as well as forage-to-concentrate ratio altered with lactation stage (i.e., dry period and early, mid, and late lactation). Diets were provided as a total mixed ration, and additional bait was fed in GreenFeed units (C-Lock Inc.), which were used for emission measurements. Supplementation of 3-NOP did not affect total DMI, BW, or BCS, but resulted in a 6.5% increase in the yields of energy-corrected milk and fat- and protein-corrected milk (FPCM). Furthermore, milk fat and protein as well as feed efficiency were increased upon 3-NOP supplementation. Overall, a reduction of 21%, 20%, and 27% was achieved for CH4 production (g/d), yield (g/kg DMI), and intensity (g/kg FPCM), respectively, upon 3-NOP supplementation. The CH4 mitigation potential of 3-NOP was affected by the lactation stage dependent diet to which 3-NOP was supplemented. On average, a 16%, 20%, 16%, and 26% reduction in CH4 yield (g/kg DMI) was achieved upon 3-NOP supplementation for the dry period, and early, mid, and late-lactation diets, respectively. The CH4 mitigation potential of 3-NOP was affected by the length of 3-NOP supplementation within a lactation stage dependent diet and by variation in diet composition within a lactation stage dependent diet as a result of changes in grass and corn silage silos. In conclusion, 3-NOP reduced CH4 emission from cows receiving 3-NOP for a year, with a positive effect on production characteristics. The CH4 mitigation potential of 3-NOP was influenced by diet type, diet composition, and nutrition value, and the efficacy of 3-NOP appeared to decline over time but not continuously. Associated with changes in diet composition, increased efficacy of 3-NOP was observed at the start of the trial, at the start of a new lactation, and, importantly, at the end of the trial. These results suggest that diet composition has a large effect on the efficacy of 3-NOP, perhaps even larger than the week of supplementation after first introduction of 3-NOP. More studies are needed to clarify the long-term effects of 3-NOP on CH4 emission and to further investigate what influence variation in diet composition may have on the mitigation potential of 3-NOP.

Effects of Fe-N co-modified biochar on methanogenesis performance, microbial community, and metabolic pathway during anaerobic co-digestion of alternanthera philoxeroides and cow manure.

The performance of anaerobic digestion (AD) is susceptible to disturbances in feedstock degradation, intermediates accumulation, and methanogenic archaea activity. To improve the methanogenesis performance of the AD system, Fe-N co-modified biochar was prepared under different pyrolysis temperatures (300,500, and 700 °C). Meanwhile, pristine and Fe-modified biochar were also derived from alternanthera philoxeroides (AP). The aim was to compare the effects of Fe-N co-modification, Fe modification, and pristine biochar on the methanogenic performance and explicit the responding mechanism of the microbial community in anaerobic co-digestion (coAD) of AP and cow manure (CM). The highest cumulative methane production was obtained with the addition of Fe-N-BC500 (260.38 mL/gVS), which was 42.37 % higher than the control, while the acetic acid, propionic acid, and butyric acid concentration of Fe-N-BC were increased by 147.58 %, 44.25 %, and 194.06 % compared with the control, respectively. The co-modified biochar enhanced the abundance of Chloroflexi and Methanosarcina in the AD system. Metabolic pathway analysis revealed that the increased methane production was related to the formation and metabolism of volatile fatty acids and that Fe-N-BC500 enhanced the biosynthesis of coenzyme A and the cell activity of microorganisms, accelerating the degradation of propionic acid and enhancing the hydrogenotrophic methanogenesis pathway. Overall, Fe-N co-modified biochar was proved to be an effective promoter for accelerated methane production during AD.

The dominant-substrate driven the enhanced performance in co-digestion of Pennisetum hybrid and livestock waste.

Co-digestion has been considered a promising method to improve methane yield. The effect of the proportion of dominant substrate on the performance and microbial community of anaerobic digestion of Pennisetum hybrid (PH) and livestock waste (LW) was investigated. An obvious synergistic effect was obtained with an increase of 15.20%-17.45% in specific methane yield compared to the predicted value. Meanwhile, the dominant substrate influenced the relational model between methane yield enhancement rate and mixture ratio. For the LW-dominant systems, a parabolic model between enhancement rate and mixture ratio was observed with a highest value of 392.16 mL/g VS achieved at a PH:LW ratio of 2:8. While a linear pattern appeared for PH-dominant systems with the highest methane yield of 307.59 mL/g VS. Co-digestion selectively enriched the relative abundance of Clostridium_sensu_stricto_1, Terrisporobacter, Syntrophomonas, Methanosarcina and Methanobacterium, which boosted the performance of hydrolysis, acidogenesis, acetogenesis and methanogenesis processes.

The resistance of hydrogenotrophic methanogenic microorganisms to ofloxacin in sludge anaerobic digestion process.

Ofloxacin (OFL) is a commonly used antibiotic that can enter wastewater treatment plants and be adsorbed by the sludge, resulting in a high OFL concentration in sludge and affecting the subsequent sludge anaerobic digestion process. However, the micro mechanisms involved in this process have not been thoroughly studied. Therefore, this study focuses on the effect of OFL on the sludge anaerobic digestion of sludge to provide such support. The experimental results showed that the maximal methane yield decreased from 277.7 to 164.7 mL/g VSS with the OFL concentration increased from 0 to 300 mg/L. Additionally, OFL hindered the intermediate biochemical processes of hydrolysis, acidogenesis, acetogenesis, and acetoclastic methanogenesis. However, it promoted hydrogenotrophic methanogenesis process, using H2 as substrate, with the concentration of 300 mg/L OFL was 5.54 fold methane production of that in the control. Further investigation revealed that the negative effect of OFL was likely due to the induction of reactive oxygen species, which led to a decrease in cell activity and interference with the activity of key enzymes. Microbiological analysis revealed that OFL reduced the relative abundance of hydrolysis and acidogenesis bacteria, and Methanosaeta archaea, while increasing the relative abundance of hydrogenotrophic methanogenesis microorganism from 36.54% to 51.48% as the OFL concentration increase from 0 to 300 mg/L.

Integrated aerobic-anaerobic digestion of highly solids-loaded corn stover and swine manure under dynamic aeration: Temperature rise, physicochemical characteristics, and methane production.

This research aimed to design an integrated aerobic-anaerobic reactor with dynamic aeration that was automatically regulated based on real-time oxygen concentration and investigate the aerobic pretreatment and subsequent dry co-anaerobic digestion (co-AD) characteristics of highly solids-loaded corn stover and swine manure in terms of temperature rise, physiochemical characteristics, and methane production. The high-temperature feedstocks from the aerobic pretreatment phase rapidly entered the AD phase without transportation and effectively improved the start-up and methane production of the co-AD. Oxygen concentration range, aeration rate, and pretreatment time affected the cumulative aeration time, temperature rise, and organic matter removal interactively during aerobic pretreatment, and a low aeration rate was relatively preferable. Although the lignocellulose removal increased with the increase in pretreatment duration, the maximal lignin elimination efficiency only reached 1.30%. The inoculum injection in the transition phase from aerobic pretreatment to co-AD and the leachate reflux during co-AD were also critical for producing methane steadily apart from aerobic pretreatment. The cold air weakened the temperature rise of aerobic pretreatment, and the low-temperature leachate reduced the methane production in the co-AD process. An oxygen concentration range of 13%-17%, aeration rate of 0.10 m3/(min·m3), pretreatment time of 84 h, inoculum loading of 40%, leachate refluxing thrice per day, and double-layer inoculation were optimum for improving the integrated aerobic-anaerobic digestion system's ability to resist low temperatures and achieving high methane production. The maximal cumulative and volatile solids (VS) methane yields of corn stover and swine manure reached 444.58 L and 266.30 L/kg VS.

Roles of quorum-sensing molecules in methane production from anaerobic digestion aided by biochar.

Biochar has been used to enhance methane generation from anaerobic digestion through establishing direct interspecific electron transfer between microorganisms. However, the microbial communication is still inadequate, thereby limiting further methane production improvement contributed by biochar. This study investigated the roles of quorum-sensing molecules, acylated homoserine lactone (AHL), in anaerobic digestion of waste activated sludge aided by biochar. Results showed that the co-addition of separated biochar and AHL achieved best methane production performance, with the maximal methane yield of 154.7 mL/g volatile suspended solids, which increased by 51.9%, 47.2%, 17.9%, and 39.4% respectively compared to that of control, AHL-loaded biochar, sole AHL, and sole biochar groups. The reason was that the co-addition of separated biochar and AHL promoted the stages of hydrolysis and acidification, promoting the conversion of organic matters and short-chain fatty acids, and optimizing the accumulation of acetate acid. Moreover, the methanogenesis stage also performed best among experimental groups. Correspondingly, the highest activities of electron transfer and coenzyme F420 were obtained, with increase ratios of 33.2% and 27.2% respectively compared to that of control. Furthermore, biochar did more significant effects on the evolution of microbial communities than AHL, and the direct interspecific electron transfer between fermentative bacteria and methanogens were possibly promoted.