Fescue toxicosis (FT) is a mycotoxin-related disease caused by the ingestion of tall fescue, naturally infected with the ergot alkaloid (EA)-producing endophyte Epichloë coenophiala. Some grazing on endophyte-free (E-) or non-toxic (NT), commercial endophyte-infected pastures takes place in the US as well. Earlier, we found that grazing on toxic fescue with low levels of EAs during fall affects thermoregulation, behavior, and weight gain. Building on these findings, the current study aimed to investigate how the presence of low EA-producing E+ or NT endophytes can influence animal metabolome, microbiome, and, ultimately, overall animal health. Eighteen Angus steers were placed on NT, E+, and E- fescue pastures for 28 days. Urine, rumen fluid (RF), rumen solid (RS), and feces were collected pre-exposure, and on days 2, 7, 14, 21, and 28. An untargeted high-resolution metabolomics approach was used to analyze urine and RF, while 16S rRNA-based next-generation sequencing (NGS) was used to examine RF, RS, feces, and fescue plant microbiomes. While alpha- or beta-microbiota diversity across all analyzed matrices were unaffected, there were specific effects of E+ on the relative abundance of some taxa (i.e., Prevotellaceae). Additionally, E+ grazing impacted aromatic amino acid metabolism in the urine and the metabolism of lipids in both the RF and urine. In both matrices, trace amine-related metabolic features differed markedly between E+ and the other groups. Compared to the endophyte-free group, endophyte presence, whether novel or toxic, influenced amino acid and carbohydrate metabolism, as well as unsaturated fatty acid biosynthesis. These findings suggest that low-EA-producing and non-toxic endophytes in fescue have more prominent effects on the metabolome than the microbiome, and this metabolome perturbation might be associated with decreased performance and reported physiological signs of FT.

Thermokarst lakes are hotspots for greenhouse gas emissions across the Arctic and Qinghai-Tibet Plateau. Investigating the vertical stratification of archaeal communities in thermokarst lake sediments is essential for understanding their ecological roles and contributions to methane production. Here, we analysed archaeal communities along a depth gradient in thermokarst lake sediments. Alpha diversity (richness and Shannon index) generally decreased with depth. Euryarchaeota was the most abundant phylum, though its relative abundance declined with depth, while Thaumarchaeota increased. At the order level, Methanosarcinales and Nitrosopumilales showed increased relative abundance with depth, indicating adaptation to deeper anoxic layers, whereas Methanomicrobiales and Methanotrichales decreased. Beta diversity increased with depth, shifting from stochastic to deterministic processes. Network topology revealed reduced species connectivity but heightened modularity at depth, signalling niche specialisation. Functionally, genes associated with the initial steps of methane metabolism (Fwd, Mtd, Mer) increased with depth, while those involved in later steps (Mtr, Mcr) decreased, suggesting reduced energy conservation efficiency and lower overall methanogenesis rates in deeper sediments. These findings highlight the significant impact of vertical stratification on archaeal community structure, interaction networks, and functional capabilities.

The limited electron transfer efficiency and poor stability of microorganisms are challenges in traditional bioelectrocatalytic food waste treatment. Herein, carbon dots (CDs) possess excellent biocompatibility and electrochemical properties. When combined with bioelectrocatalysis, CDs can regulate microbial community structure and enhance electron exchange capacity. The results demonstrated that compared with the control group (28.1 mL/g-VS/d), the CDs at a dosage of 0.50 g/g VS could increase CH4 production by a factor of 7.8. CDs not only increase CH4 production but also improve the digestate's stability, making it suitable for use as bio-fertilizer. Moreover, a significantly high Methanobacterium richness (11.6 %) signified an intensified utilization of hydrogen and formic acid pathways in CH4 production. Particularly, the biocompatible CDs could be absorbed by microorganisms, forming an environmental network that was more conducive to electron transfer with unabsorbed CDs and accelerating interspecies electron transfer. This work provides mechanistic insights into boosting CH4 production in AD.

The effect of the dietary inclusion of Hermetia illucens larvae meal on the diversity of the methanogenic archaea in the caecum of laying hens (Hy-line Brown) was investigated using molecular methods. A total of 27 hens, selected equally for slaughter from 162 birds which were divided equally into 3 treatment groups including control group C with a diet containing corn-soybean meal and 2 experimental groups, HI25 and HI50, in which 25% and 50% of the soybean meal protein was replaced by the protein from a Hermetia illucens larvae meal, respectively. At 40 weeks of age, the methanogenic community of caecal content of 9 hens per group was analyzed using a 16S rRNA gene clone library. A total of 108 positive clones, 35 from the control group, 44 from the HI25 group and 29 from the HI50 group, were analyzed by Sanger sequencing. Methanomicrobiales, Methanobacteriales and Methanomassiliicoccales were the main orders found in groups C and HI25. Methanomassiliicoccales was absent in the HI50 group, which was dominated by the order Methanobacteriales. At the species level, Methanobrevibacter woesei was the most prevalent species in all three groups regardless of diet. Some species were found exclusively either in the control group (Methanogenic archaeon CH1270) or in the HI25 group (Methanorbis furvi strain Ag1). Methanogenic diversity was significantly lower in the HI50 group compared to the control and HI25 groups and Methanomassiliicoccaceae archaeon DOK was completely suppressed in HI50 group. Our preliminary results indicate that ingestion of Hermetia illucens larvae meal has considerable effect on the methanogenic community, promoting the abundance of Methanobrevibacter woesei and suppressing Methanomassiliicoccaceae archaeon DOK in the caeca of laying hens.

Members of the phylum Synergistota are important but understudied components of microbial communities during anaerobic digestion. In this study, their diversity was assessed in full-scale anaerobic digester sludge samples from Marrakesh wastewater treatment plant (Morocco), using 16S rRNA gene community profiling, as well as targeted isolation, physiological characterization, and genome sequencing of novel Synergistaceae isolates. The 16S rRNA gene analysis identified 23 operational taxonomic units (OTUs) belonging to the family of Synergistaceae, representing 8.8 % of the total microbial community. 17 of these OTUs belonged to previously uncultured taxa. A dominant OTU19, presumably a new representative of the family of Synergistaceae was isolated in pure culture (strain DS-S4T) and subjected to both culture- and genome-based characterizations. Phylogenetic analysis revealed that strain DZ-S4T was related to Cloacibacillus porcorum CL-84T and Cloacibacillus evryensis 158T but with low sequence similarity of 89.94 % and 88.60 %, respectively. Based on genome relatedness, including Average Nucleotide Identity (ANI) and Amino Acid Identity (AAI), strain DZ-S4T is considered to represent a novel genus for which the name Caenicola gen.nov is proposed. Moreover, several phenotypic and eco-physiological properties differentiated the novel isolate from its related species, indicating that the strain represents a new species for which the name Caenicola nitritireducens sp. nov. is proposed, with strain DZ-S4T (=DSM 104940T = JCM 31897T) being the type strain. Additionally, this study investigates the ecological role of strain DZ-S4T, specifically the protein degradation, the bioconversion of carbohydrates, and the nitrite reduction during anaerobic digestion.

Inland rivers play a crucial role in regulating the methane (CH4) budget via microbial carbon cycling. CH4 emissions vary significantly among rivers with different land covers, yet the regulatory mechanisms of CH4-cycling microorganisms across different land covers remain less understood. This study intergrates in-situ CH4 measurements with amplicon and metagenomic sequencing to investigate CH4-cycling microbial community composition and metabolic functions in regulating CH4 emissions across rivers with different land covers. Our results show that agricultural and urban rivers significantly increase riverine CH4 emission fluxes by 14 and 34 times than forest rivers, respectively. Urban and agricultural river sediments exhibited higher methanogenic abundance, but lower methanotrophic abundance than forest river sediments. Acetoclastic methanogens dominate the methanogenic communities of agricultural rivers, enhanced by high NO3- and DOC concentrations. Furthermore, methanogenic metagenome-assembled genomes (MAGs) recovered from agricultural rivers, which affiliated to Methanosarcina, carried the complete set of genes encoding for the enzymes in acetoclastic methanogenesis. In contrast, hydrogenotrophic methanogens drive CH4 production in urban rivers, favored by low DOC: NH4+ ratios that enable methanogenesis independent of organic carbon. Lower CH4 emissions in agricultural rivers compared to urban rivers might be partly due to the greater sulfate-dependent anaerobic methane-oxidation. In forest rivers, type I methanotrophs outcompetes methanogens, aided by suitable sediment pH and larger sediment particle sizes, fostering oxic conditions that suppress CH4 emissions. This study reveals versatile microbial mechanisms underlying riverine CH4 emissions across land covers, enhancing understanding of microbial-mediated riverine CH4 cycling.

Glacier-fed streams (GFS) are emergent sources of greenhouse gas methane, and methanogenic archaea in sediments contribute largely to stream methane emissions. However, little is known about the methanogenic communities in GFS sediments and their key environmental driving factors. This study analyzed stream sediments from the Rongbuk River basin on Mt. Everest for methanogenic communities and their temperature responses through anaerobic microcosm incubations at 5°C and 15°C. Diverse methanogens were identified, including acetoclastic, hydrogenotrophic, and hydrogen-dependent methylotrophic types. Substantial methane and CO2 production were detected across altitudes and increased significantly at 15°C, with both methane and CO2 production rates negatively correlated with altitude. The temperature sensitivity of CO2 production also showed a negative altitude correlation. Methanogens increased substantially over long-term incubation, dominating the archaeal community. At 15°C, the relative abundance of several methanogenic groups was strongly correlated with altitude, with positive correlations observed for Methanomassiliicoccaceae and Methanoregulaceae, and negative correlations for Methanocellaceae, respectively. Besides altitude, phosphorus, carbon to nitrogen ratio, and pH also affected methanogenic structure, methane and CO2 production, and temperature sensitivities. This study offers new insights into methanogens and methane production in GFS sediments, improving our understanding of GFS carbon cycling and its potential responses to climate change.

Numerous cultivation techniques for aerobic microorganisms have been extensively investigated in the field of microbiology. Optimisation of these techniques is important for scientific and economic reasons. Methanogenic archaea, however, are obligate anaerobic microorganisms requiring different cultivation techniques than aerobic organisms due to the fundamental differences in physiology. Mixing of aerobic cultures is generally considered as very important as it provides organisms with essential oxygen; however, for anaerobic microorganisms lacking the ability to grow with oxygen, this point in cultivation was widely neglected. This work aimed at investigating the effect of different mixing modes on cultures of the methanogenic archaea Methanomethylovorans thermophila, Methanosarcina acetivorans, Methanosarcina thermophila and Methanococcus vannielii by cultivating them anaerobically in the modes standing/lying, shaken/unshaken and large/small serum flask in order to analyse their impact on the methane and biomass production. This study showed that a shaken incubation mode had a positive impact on methane production and resulted in its accelerated production, especially in hydrogenotrophic cultures; however, higher methane production did not necessarily lead to higher biomass production.

The in situ biomethanation process aims to increase the CH4 content in biogas by injecting H2 into anaerobic digesters, improving its energetic value. However, H2 injection causes CO2 depletion hampering in situ biomethanation performances. This work investigated the effect of H2 addition and CO2 depletion on the adaptation of two microbial consortia to in situ biomethanation. In the in situ biomethanation reactors under CO2-non-limiting conditions (biogas CO2 concentration ∼ 18 %) CH4 was the most produced metabolite (∼80 % of the produced gCOD) whatever the microbial consortia. However, in the in situ biomethanation reactors under CO2 limitation (biogas CO2 concentration ∼ 3 %), CH4 and Volatile Fatty Acids (VFA) accounted for 60 and 40 % of the produced gCOD, despite the tested microbial consortia. Interestingly, all control reactors (operated without H2 and/or CO2 addition) produced mostly VFA instead of CH4 (∼70 to 30 % COD-based-proportion). Hence, VFA accumulation was alleviated by H2 injections in both in situ biomethanation conditions. This lower VFA accumulation was associated with the adaptation of the microbial consortia to H2, evidenced by the improved growth of hydrogenotrophic methanogens (HM). Moreover, competition between HM clades may play a role in microbial adaptation to H2. However, low CO2 availability hindered HM growth and led to lower VFA conversion to CH4 in the in situ biomethanation conditions under CO2 limitation. Methanobacterium spp. was highly resistant to CO2 depletion, dominating the archaeal community in these conditions. This study demonstrated that the microbial adaptation to H2 addition, characterised by an improved HM activity, boosted methanogenesis and enhanced indirectly acetogenesis, preventing VFA accumulation even under CO2-depleted conditions.

Black and odorous waters (BOWs) are a serious environmental problem frequently reported over the past few decades. Microorganisms are identified as implementors of the black and odorous phenomenon, which play a crucial role in the decomposition and transformation of pollutants within the BOWs. However, the information on the role of microorganisms in BOWs remains elusive. BOWs are characterized by high concentrations of organic compounds and limited oxygen inputs, which have facilitated the emergence of distinct microbial species. The algae, hydrolytic and fermentative bacterium, sulfate-reducing bacteria, Fe-reducing bacteria and other microorganisms play an important role in the process of blackening and odorization of waters. Studying these specific microbial taxonomies provides valuable insights into their adaptations and contributions to the overall functioning of BOWs. This study comprehensively reviews 1) the microbial community structure, assembly and succession in BOWs; 2) the key microbial profiles involved in BOWs formation; 3) the interspecies interactions process in the BOWs, which are the issues easily overlooked but deserve further research and development.

Myostatin (MSTN) is a protein that plays a crucial role in regulating skeletal muscle development. Despite the known benefits of MSTN mutant cattle for increasing beef production, their potential impact on CH4 emissions has not been quantified. The study comparing wild-type (WT) cattle to MSTN-knockout (MSTN-KO) cattle revealed that CH4 production was lower. Macrogenomic analysis revealed a significant decrease in rumen archaea, with reduced Richness indices (P = 0.036). The MSTN-KO cattle also showed altered archaea distribution and composition at different taxonomic levels. LEfSe results showed changes in 21 methanogenic archaea clades, with obligately hydrogen (H2)-dependent methylotrophs Candidatus Methanoplasma termitum species belonging to Methanomassiliicoccales order demonstrating the most significant decrease. Rumen metabolites revealed a decrease in the ratio of acetate to propionate, indicating a shift in rumen fermentation pattern towards propionate fermentation. Additionally, the changing trend of methanogenic archaea is consistent with the evolution of methanogens, and this is correlated with the higher levels of linoleic acid in the rumen of MSTN-KO cattle. Linoleic acid affects the utilization of H2 by methanogenic archaea, leading to a reduction in obligately H2-dependent methylotrophs. Our study suggests that MSTN-KO cattle have potential as an economically and ecologically benign breed for reducing methane emissions.

In soil polluted with benzene, toluene, ethylbenzene, and xylenes (BTEX), oxygen is rapidly depleted by aerobic respiration, creating a redox gradient across the plume. Under anaerobic conditions, BTEX biodegradation is then coupled with fermentation and methanogenesis. This study aimed to characterize this multi-step process, focusing on the interactions and functional roles of key microbial groups involved. A reactor system, comprising an Anaerobic Bioreactor (AB) and two Microbial Electrolysis Cell (MEC) chambers, designed to represent different spatial zones along the redox gradient, operated for 160 days with intermittent exposure to BTEX. The functional differentiation of each chamber was reflected by the gas emission profiles: 50%, 12% and 84% methane in the AB, anode and cathode chambers, respectively. The taxonomic profiling, assessed using 16S amplicon sequencing, led to the identification chamber-characteristic taxonomic groups. To translate the taxonomic shift into a functional shift, community dynamics was transformed into a simulative platform based on genome scale metabolic models constructed for 21 species that capture both key functionalities and taxonomies. Representatives include BTEX degraders, fermenters, iron reducers acetoclastic and hydrogenotrophic methanogens. Functionality was inferred according to the identification of the functional gene bamA as a biomarker for anaerobic BTEX degradation, taxonomy and literature support. Comparison of the predicted performances of the reactor-specific communities confirmed that the simulation successfully captured the experimentally recorded functional variation. Variations in the predicted exchange profiles between chambers capture reported and novel competitive and cooperative interactions between methanogens and non-methanogens. Examples include the exchange profiles of hypoxanthine (HYXN) and acetate between fermenters and methanogens, suggesting mechanisms underlying the supportive/repressive effect of taxonomic divergence on methanogenesis. Hence, the platform represents a pioneering attempt to capture the full spectrum of community activity in methanogenic hydrocarbon biodegradation while supporting the future design of optimization strategies.

Marine sediments contain Earth's largest reservoir of methane, with most of this methane being produced and consumed in situ by methane-cycling archaea. While numerous studies have investigated communities of methane-cycling archaea in hydrocarbon seeps and sulfate-methane transition zones, less is known about how these archaea change from the seafloor downward throughout diffusion-dominated marine sediments. Focusing on four continental margin sites of the North Sea-Baltic Sea transition, we here investigate the in situ drivers of methane-cycling archaeal community structure and metabolism based on geochemical and stable carbon-isotopic gradients, functional gene (mcrA) copy numbers and phylogenetic compositions, and thermodynamic calculations. We observe major changes in community structure that largely follow vertical gradients in sulfate concentrations and lateral gradients in organic carbon reactivity and content. While methane-cycling archaeal communities in bioturbated and sulfatic zones are dominated by known methyl-disproportionating Methanosarcinaceae and putatively CO2-reducing Methanomicrobiaceae, the communities change toward dominance of methane-oxidizing taxa (ANME-2a-b, ANME-2c, ANME-1a-b) in sulfate-methane transition zones (SMTZs). By contrast, the underlying methanogenesis zones are dominated by the physiologically uncharacterized ANME-1d, new genus-level groups of putatively CO2-reducing Methanomicrobiaceae, and methyl-reducing Methanomassiliicoccales. Notably, mcrA copy numbers of several major taxa increase by 2 to 4 orders of magnitude from the sulfatic zone into the SMTZ or methanic zone, providing evidence of net population growth in subsurface sediment. We propose that burial-related geochemical changes cause methane-cycling archaea in continental margin sediments to go through three successional stages (sulfatic, SMTZ, methanic). Herein, the onset of each new successional stage is characterized by a period of growth- and mortality-driven turnover in the dominant taxa.

Polyhydroxyalkanoate (PHA) is the important biodegradable plastic, however, biodegradation of PHA waste in anaerobic environments emits more CH4, a potent greenhouse gas. Bioconversion of PHA waste to useful byproducts - volatile fatty acids (VFAs) is a practical method to upcycle carbon from PHA. In this study, PHA waste was anaerobically co-digested with swine manure (SM) (the typical high nitrogen waste) at different C/N ratios. The results indicate that co-digestion of PHA and SM with a C/N ratio of 32.1 achieved VFA production of 5488 mg COD/L and 0.20 g COD/g VS. No significant differences were found in terms of the highest VFA concentrations between treatments with C/N ratios of 43.4 and 32.1. VFA produciton of 3655 mg COD/L and 0.14 g COD/g VS was achieved at 19 days by adjusting the C/N ratio to 19.2. Four bacteria were identified as dominant microorganisms responsible for converting PHA and SM to VFA.

The biochemical composition of sediments, which depends on the origin of the organic matter (OM), is decisive in methane (CH4) production. This study aimed to determine the CH4 produced under anaerobic conditions from different substrates: native reservoir sediments and sediments with the addition of complex OM from Microcystis spp. blooms and terrestrial plants (pasture), alongside the biochemical characterization of the substrates used. The biochemical composition of the sediments explained the differences in CH4 production rate (µmol/g OM/day). Positive correlations were found between CH4 production rate and proteins (PRT) (r = 0.695), lipids (LIP) (r = 0.582), TN (r = 0.605), and biopolymeric carbon (BPC) (r = 0.784). Principal component analysis showed an association of CH4 production rate with PRT, LIP, TN, and BPC concentrations, sharing the same direction of the vectors. The addition of Microcystis bloom and pasture to the sediments significantly increased the rate and production of CH4 compared to native sediments, with higher values for bloom addition. In the studied reservoirs, cyanobacteria biomass is a very important source of organic matter to the sediments and a support for methanogenesis. Nonetheless, OM from surrounding vegetation, mainly pastures, could also play a significant role during events that increase reservoir levels, generating important CH4 emission "hot spots" at the periphery of the reservoirs.

This study assessed the impact of organic loading rate (OLR) on methane (CH4) production in the anaerobic co-digestion (AcoD) of sugarcane vinasse and molasses (SVM) (1:1 ratio) within a thermophilic fluidized bed reactor (AFBR). The OLR ranged from 5 to 27.5 kg COD.m-3.d-1, with a fixed hydraulic retention time (HRT) of 24 h. Organic matter removal varied from 56 to 84%, peaking at an OLR of 5 kg COD.m-3.d-1. Maximum CH4 yield (MY) (272.6 mL CH4.g-1CODrem) occurred at an OLR of 7.5 kg COD.m-3.d-1, while the highest CH4 production rate (MPR) (4.0 L CH4.L-1.d-1) and energy potential (E.P.) (250.5 kJ.d-1) were observed at an OLR of 20 kg COD.m-3.d-1. The AFBR exhibited stability across all OLR. At 22.5 kg COD.m-3.d-1, a decrease in MY indicated methanogenesis imbalance and inhibitory organic compound accumulation. OLR influenced microbial populations, with Firmicutes and Thermotogota constituting 43.9% at 7.5 kg COD.m-3.d-1, and Firmicutes dominating (52.7%) at 27.5 kg COD.m-3.d-1. Methanosarcina (38.9%) and hydrogenotrophic Methanothermobacter (37.6%) were the prevalent archaea at 7.5 kg COD.m-3.d-1 and 27.5 kg COD.m-3.d-1, respectively. Therefore, this study demonstrates that the organic loading rate significantly influences the efficiency of methane production and the stability of microbial communities during the anaerobic co-digestion of sugarcane vinasse and molasses, indicating that optimized conditions can maximize energy yield and maintain methanogenic balance.

As an accelerated electron transfer device, the influence of microbial electrochemical snorkel (MES) on soil greenhouse gas production remains unclear. Electron transport is the key to methane production and denitrification. We found that the N2O amount of the MES treatment was comparable to the control however the cumulative CO2 and CH4 emissions were reduced by 50% and 41%, respectively. The content of Fe2+ in MES treatment increased by 31%, which promoted the electron competition of iron reduction to methanogenesis. Furthermore, the competition among iron-reducing, nitrifying and denitrifying bacteria reduced the abundance of methanogens by 19-20%. Additionally, the MES treatment decreased the abundance of genes associated with hydrogen methanogenesis pathway by 6-19%, and inhibited the further conversion of acetyl-CoA into CH4 for acetoclastic methanogenesis. This study reveals effects of accelerating electron transfer on greenhouse gas emission, and provides a novel strategy for reducing greenhouse gas emissions in paddy soil.

The biomethanation process is widely recognized as a significant approach to mitigating carbon dioxide emissions while simultaneously generating methane. However, only a few microorganisms that required intricate culturing conditions were identified for biomethanation. Here, Escherichia coli that featured easy cultivation and versatile chassis was genetically modified for biomethanation for the first time. The nif-related gene cluster with methanogenic capability from Rhodopseudomonas palustris was systematically analyzed, cloned, and integrated into a synthetic biomethanation module. As a result, E. coli BL21 (DE3) and Rosetta (DE3) carrying this synthetic biomethanation module exhibited significant methane production activity, with methane yields reaching 50 nmol/mL and 159 nmol/mL, respectively. This finding provided a simple route to construct synthetic strain for biomethanation, which would advance the fundamental research and be beneficial to further harness the power of biomethanation for practical application.

Ruminants play an important role in global food security and nutrition. The rumen microbial community provides ruminants with a unique ability to convert human indigestible plant matter, into high quality edible protein. However, enteric CH4 produced in the rumen is both a potent GHG and a metabolizable energy loss for ruminants. As the rumen microbiome constitutes 15-40% of the inter-animal variation in enteric CH4 emissions, understanding the microbiological mechanisms underpinning ruminal methanogenesis and its interaction with the host animal, is crucial for developing CH4 mitigation strategies. Variation in the relative abundance of different microbial species has been observed in cattle with contrasting residual CH4 emission and CH4 yield with up to 20% of the variation in inter-animal CH4 emissions attributable to the presence of a small number of microbial species. The demonstration of ruminotypes associated with high or low CH4 emissions suggests that interactions within complex microbial consortia and with their host are a major source of variation in CH4 emissions. Consequently, microbiome-assisted genomic approaches are being developed to select low CH4 emitting cattle, with breeding values for enteric CH4 being included as part of national breeding programmes. Generating rumen microbiome data for use in selection programs is expensive, therefore, identifying microbial biomarkers in milk or plasma to develop predictive models which include microbial predictors in equations based on animal related data, is required. A better understanding of the rumen microbiome has also aided the development and refinements of anti-methanogenic feed additives. However, these strategies, which increase the amount of reducing equivalents in the rumen ecosystem, do not generally result in an enrichment of propionate or an improvement in animal performance. Current research aims to provide alternative sinks to reducing equivalents and to stimulate activity of commensal microbes or the supplementation of direct fed microbials to capture lost energy. Furthering our knowledge of the rumen microbiome and its interaction with the host, will aid in the development of CH4 mitigation strategies for ruminant livestock.

Current estimates of wetland contributions to the global methane budget carry high uncertainty, particularly in accurately predicting emissions from high methane-emitting wetlands. Microorganisms drive methane cycling, but little is known about their conservation across wetlands. To address this, we integrate 16S rRNA amplicon datasets, metagenomes, metatranscriptomes, and annual methane flux data across 9 wetlands, creating the Multi-Omics for Understanding Climate Change (MUCC) v2.0.0 database. This resource is used to link microbiome composition to function and methane emissions, focusing on methane-cycling microbes and the networks driving carbon decomposition. We identify eight methane-cycling genera shared across wetlands and show wetland-specific metabolic interactions in marshes, revealing low connections between methanogens and methanotrophs in high-emitting wetlands. Methanoregula emerged as a hub methanogen across networks and is a strong predictor of methane flux. In these wetlands it also displays the functional potential for methylotrophic methanogenesis, highlighting the importance of this pathway in these ecosystems. Collectively, our findings illuminate trends between microbial decomposition networks and methane flux while providing an extensive publicly available database to advance future wetland research.

Seagrass meadows are acknowledged as blue carbon ecosystems, yet they are also ideal habitats for methane (CH4) release, offsetting their ability to mitigate climate change. The global CH4 fluxes in seagrass meadows remain highly uncertain due to regional and species biases, and the microbial mechanisms driving methane release are poorly understood. Here, we investigated CH4 air-sea fluxes, sediment CH4 emission potential and microbes involved in CH4 release using geochemical techniques combined with qPCR and Illumina sequencing in a temperate Zostera japonica and Zostera marina mixed meadow. The CH4 air-sea fluxes fluctuated from -0.42 to 11.42 μmol·m-2·d-1, showing a strong seasonal variation. CH4 emission potential was significantly higher in seagrass vegetated sediments (10.34 ± 2.72 nmol·g-1·d-1) than in the adjacent bare sediments (1.55 ± 1.15 nmol·g-1·d-1), primarily attributed to variations in sediment organic matter content. Diverse methanogens occurred in the seagrass meadow, with Methanolobus dominating in seagrass sediments, while Methanococcoides, Methanosarcina, and Methanoculleus being prevalent in bare sediments. Meanwhile, a variety of methylotrophic groups were detected, including aerobic Gammaproteobacteria, anaerobic Desulfobacterota and Methylomirabilota, as well as archaea Candidatus Methanoperedens. The co-occurrence of these functional groups implied the presence of complex CH4 production and oxidation pathways, which regulated the CH4 budget in the seagrass ecosystems. Taken together, our findings enhance the comprehension of the methane emission process and driving mechanism in seagrass ecosystems.

This publication aims to provide guidelines of the knowledge required and the potential research to be conducted in order to understand the mode of action of antimethanogenic feed additives (AMFA). In the first part of the paper, we classify AMFA into 4 categories according to their mode of action: (1) lowering dihydrogen (H2) production; (2) inhibiting methanogens; (3) promoting alternative H2-incorporating pathways; and (4) oxidizing methane (CH4). The second part of the paper presents questions that guide the research to identify the mode of action of an AMFA on the rumen CH4 production from 5 different perspectives: (1) microbiology; (2) cell and molecular biochemistry; (3) microbial ecology; (4) animal metabolism; and (5) cross-cutting aspects. Recommendations are provided to address various research questions within each perspective, along with examples of how aspects of the mode of action of AMFA have been elucidated before. In summary, this paper offers timely and comprehensive guidelines to better understand and reveal the mode of action of current and emerging AMFA.

Biochar is a promising strategy to solve the problem of low efficiency and ammonia inhibition during anaerobic digestion (AD). However, the correlation between biochar partitioning and its stimulatory effects on AD remains uncertain. Here, the effects of partitioned Ganoderma lucidum residue biochar (GLRB) on biogas and methane production were investigated. The GLRB produced at 450 °C, with richer functional groups on its surface, had the optimal enhancement effect on AD, resulting in a 20.59% increase in methane production compared with control. The doses of water-soluble GLRB (LZ450-W) and water-insoluble GLRB (LZ450-R) were not proportional to their enhancement effect on AD. However, the enhancement effect on AD by LZ450-R was better than that of LZ450-W. The optimal dosage of LZ450-W was 0.015 g, which increased methane production by 14.28%. Similarly, methane production increased by 26.91% with the addition of 0.603 g of LZ450-R. LZ450-R had more abundant functional groups on the surface and promoted the abundance of bacteria in the dominant phyla Bacteroidetes, Synergistetes, and Spirochaetes, increasing the rate of hydrolysis. Additionally, methanogens such as Methanobacterium and Methanospirillum were enriched, facilitating methane production by promoting the hydrogenotrophic pathway. Methanobacterium was also negatively correlated with most acid-producing bacteria, whereas Methanobrevibacter was positively correlated with Methanosphaera, Acetivibrio, and other acid-producing bacteria. These findings provide a basis for constructing synthetic microbial communities using biochar as a carrier of microbial inoculum.

Safe geological disposal of radioactive waste requires a thorough understanding of geochemical conditions in the host formation. Boom Clay is a potential candidate in Belgium, where active methanogenesis has been detected in its deep subsurface, influencing the local geochemistry. However, the pathways driving this process and the characteristics of the methanogenic archaea involved remain unclear. We isolated a distinct archaeal strain from Boom Clay pore water and characterized it geno- and phenotypically. Isolate TD41E1-1 belongs to a novel species of the Methanosarcina genus, for which the name Methanosarcina hadiensis sp. nov. is proposed. TD41E1-1 cells are coccus-shaped, irregularly sized cells enveloped by extracellular polymer substances. Growth and substrate utilization experiments and genomic analysis demonstrated that the strain prefers methylated compounds or hydrogen as substrates for methane production. Although it possesses a complete acetoclastic pathway, no growth was observed in the presence of acetate in the tested conditions. Based on its phylogenetic relation to other known Methanosarcina species and on the presence of c-type cytochromes, it can be concluded that the strain likely occupies an intermediate position between type I and type II Methanosarcina species. These findings provide valuable insights for assessing Boom Clay's suitability for geological disposal of radioactive waste.

Aquaculture pond sediments act as hotspots for methane (CH4) emissions; however, knowledge gaps on the regulation of microorganisms hinder our further understanding of methane dynamics in aquaculture pond sediment. Using field sampling and molecular analysis, we examined CH4 fluxes, the methanogenic community composition, and their interaction with methanotrophs to comprehensively understand the methane cycling in sediments of aquaculture ponds in northern China. Compared with a fishing pond without feed inputs, the abundances of methanogens mcrA and methanotrophs pmoA genes increased significantly in aquaculture ponds sediments. The dominant methanogens were Methanothrix, Methanoregula, and Methanolinea, and the α-diversity indices of methanogens demonstrated higher levels in 0-5 cm surface sediment. The methanotrophs were dominated by Methylocystis, Methylocaldum, and Methylobacter, and the α-diversity indices of methanotrophs showed no significant difference. The total organic carbon (TOC) contents and oxidation reduction potential (ORP) were the key factors driving methanogenic and methanotrophic communities on methane cycle in aquaculture sediment. The inter-domain ecological network (IDEN) analysis revealed that total number of network nodes, links, connectances, and links per species in the aquaculture sediments presented relatively higher levels, whereas the IDEN modules were fewer. The methanogens dominated in the networks and the interaction of methanogens and methanotrophs was more competitive and complex in aquaculture sediments. These findings highlight the marked methane production in aquaculture sediment, primarily due to the abundance, diversity, and competitive advantage of methanogens over methanotrophic communities.

Although mangrove forests can uptake atmospheric CO2 and store carbon as organic matter called "blue carbon", it is also an important natural source of greenhouse gas methane. Methanogens are major contributors to methane and play important roles in the global carbon cycle. However, our understanding of the key microbes and metabolic pathways responsible for methanogenesis under specific substrates in mangrove sediments is still very limited. Here, we set an anaerobic incubation to evaluate the responses of methanogens in mangrove sediments from South China to the addition of diverse methanogenic substrates (H2/CO2, acetate, trimethylamine (TMA), and methanethiol (MT)) and further investigated the dynamics of the whole microbial community. Our results showed that diverse substrates stimulated methanogenic activities at different times. The stimulation of methanogenesis was more pronounced at early and late periods by the addition of methylotrophic substrates TMA and MT, respectively. The amplicon sequencing analysis showed that genus Methanococcoides was mainly responsible for TMA-utilized methanogenesis in mangrove sediment, while the multitrophic Methanococcus was most abundant in H2/CO2 and MT treatments. Apart from that, the bacteria enrichments of Syntrophotalea, Clostridium_sensu_stricto_12, Fusibacter in MT treatments might also be associated with the stimulation of methane production. In addition, the metagenomic analysis suggested that Methanosarcinaceae was also one of the key methanogens in MT treatments with different genomic information compared to that in TMA treatments. Finally, the total relative abundances of methanogenesis-related genes were also highest in TMA and MT treatments. These results will help advance our understanding of the contributions of different methanogenesis pathways and methanogens to methane emissions in mangrove sediments.

Molecular hydrogen is produced by the fermentation of organic matter and consumed by organisms including hydrogenotrophic methanogens and sulfate reducers in anoxic marine sediment. The thermodynamic feasibility of these metabolisms depends strongly on organic matter reactivity and hydrogen concentrations; low organic matter reactivity and high hydrogen concentrations can inhibit fermentation so when organic matter is poor, fermenters might form syntrophies with methanogens and/or sulfate reducers who alleviate thermodynamic stress by keeping hydrogen concentrations low and tightly controlled. However, it is unclear how these metabolisms effect porewater hydrogen concentrations in natural marine sediments of different organic matter reactivities.

We measured aqueous concentrations of hydrogen, sulfate, methane, dissolved inorganic carbon, and sulfide with high-depth-resolution and 16S rRNA gene assays in sediment cores with low carbon reactivity in White Oak River (WOR) estuary, North Carolina, and those with high carbon reactivity in Cape Lookout Bight (CLB), North Carolina. We calculated the Gibbs energies of sulfate reduction and hydrogenotrophic methanogenesis.

Hydrogen concentrations were significantly higher in the sulfate reduction zone at CLB than WOR (mean: 0.716 vs. 0.437 nM H2) with highly contrasting hydrogen profiles. At WOR, hydrogen was extremely low and invariant (range: 0.41-0.52 nM H2) in the upper 15 cm. Deeper than 15 cm, hydrogen became more variable (range: 0.312-2.56 nM H2) and increased until methane production began at ~30 cm. At CLB, hydrogen was highly variable in the upper 15 cm (range: 0.08-2.18 nM H2). Ratios of inorganic carbon production to sulfate consumption show AOM drives sulfate reduction in WOR while degradation of organics drive sulfate reduction in CLB.

We conclude more reactive organic matter increases hydrogen concentrations and their variability in anoxic marine sediments. In our AOM-dominated site, WOR, sulfate reducers have tight control on hydrogen via consortia with fermenters which leads to the lower observed variance due to interspecies hydrogen transfer. After sulfate depletion, hydrogen accumulates and becomes variable, supporting methanogenesis. This suggests that CLB's more reactive organic matter allows fermentation to occur without tight metabolic coupling of fermenters to sulfate reducers, resulting in high and variable porewater hydrogen concentrations that prevent AOM from occurring through reverse hydrogenotrophic methanogenesis.

Enhanced weathering of (ultra)mafic rocks has been proposed as a promising approach to sequester atmospheric CO2 and mitigate climate change. However, these silicate rocks contain varying amounts of trace metals, which are essential cofactors of metallaenzymes in methanogens. We found that weathering of crushed peridotite and basalt significantly promoted the growth and methanogenesis of a model methanogen─Methanosarcina acetivorans C2A under the condition of excess substrate. The released trace metals from peridotite and basalt, especially Fe, Ni, and Co, accounted for the promotion effect. Observation at different spatial scales showed a close association between the rocks and cells. Proteomic analysis revealed that rock amendment significantly enhanced the expression of core metalloenzymes in the methylotrophic methanogenesis pathway. Our study uncovers a previously unrecognized but important negative effect of enhanced rock weathering on methane production, which may counteract the carbon sequestration effort.

A high rate upflow anaerobic polyfoam-based bioreactor (UAPB) was developed for lab-scale in-situ biogas upgrading by H2 injection. The reactor, with a volume of 440 mL, was fed with synthetic wastewater at an organic loading rate (OLR) of 3.5 g COD/L·day and a hydraulic retention time (HRT) of 7.33 h. The use of a porous diffuser, alongside high gas recirculation, led to a higher H2 liquid mass transfer, and subsequently to a better uptake for high CH4 content of 56% (starting from 26%). Our attempts to optimize both operational parameters (H2 flow rate and gas recirculation ratio, which is the total flow rate of recirculated gas over the total outlet of gas flow rate) were not initially successful, however, at a very high recirculation ratio (32) and flow rate (54 mL/h), a significant improvement of the hydrogen consumption was achieved. These operational conditions have in turn driven the methanogenic community toward the dominance of Methanosaetaceae, which out-competed Methanosarcinaceae. Nevertheless, highly stable methane production rates of 1.4-1.9 L CH4/Lreactor.day were observed despite the methanogenic turnover. During the different applied operational conditions, the bacterial community was especially impacted, resulting in substantial shifts of taxonomic groups. Notably, Aeromonadaceae was the only bacterial group positively correlated with increasing hydrogen consumption rates. The capacity of Aeromonadaceae to extracellularly donate electrons suggests that direct interspecies electron transfer (DIET) enhanced biogas upgrading. Overall, the proposed innovative biological in-situ biogas upgrading technology using the UAPB configuration shows promising results for stable, simple, and effective biological biogas upgrading.

Fungi inhabiting deep subseafloor sediments have been shown to possess anaerobic methane (CH4) production capabilities under atmospheric conditions. However, their ability to produce CH4 under in situ conditions with high hydrostatic pressure (HHP) remains unclear. Here, Schizophyllum commune 20R-7-F01, isolated from ~2 km below the seafloor, was cultured in Seawater Medium (SM) in culture bottles fitted with sterile syringes for pressure equilibration. Subsequently, these culture bottles were transferred into 1 L stainless steel pressure vessels at 30 °C for 5 days to simulate in situ HHP and anaerobic environments. Our comprehensive analysis of bioactivity, biomass, and transcriptomics revealed that the S. commune not only survived but significantly enhanced CH4 production, reaching approximately 2.5 times higher levels under 35 MPa HHP compared to 0.1 MPa standard atmospheric pressure. Pathways associated with carbohydrate metabolism, methylation, hydrolase activity, cysteine and methionine metabolism, and oxidoreductase activity were notably activated under HHP. Specifically, key genes involved in fungal anaerobic CH4 synthesis, including methyltransferase mct1 and dehalogenase dh3, were upregulated 7.9- and 12.5-fold, respectively, under HHP. Enhanced CH4 production under HHP was primarily attributed to oxidative stress induced by pressure, supported by intracellular reactive oxygen species (ROS) levels and comparative treatments with cadmium chloride and hydrogen peroxide. These results may provide a strong theoretical basis and practical guidance for future studies on the contribution of fungi to global CH4 flux.

High solid anaerobic digestion has proved the mainstream technology for the treatment of organic wastes. However, the high molecular weight and complex lignocellulosic structure of cow manure (CM) make it indigestible and inefficient, leading to limit the hydrolysis step of anaerobic digestion at high solid content. To mitigate this bottleneck, an improved cost-effective freezing and thawing pretreatment technique was proposed in this study. The freezing and thawing pretreatment of raw CM without any dilution was done for 20 days. The maximum cumulative methane yield (487 mL CH4 g- 1VS) was achieved at a total solid (TS) of 5% followed by TS of 10% and 15%, which was 13%, 20% and 21% higher than obtained from untreated CM, respectively. The kinetic results revealed that the biodegradable materials could be utilized at increasing TS with decreasing hydrolysis rate. The pretreatment significantly enhanced the methylotrophic methanogenic pathway during high solid anaerobic digestion, which was contrary to the general concept that the process is usually dominated by acetoclastic and hydrogenotrophic methanogens. This study is very important to understand the effect of solid content but also important to understand the effect of freezing and thawing pretreatment on process parameters and microbial community dynamics in high solid anaerobic digestion.

Anaerobic digestion is one of the most promising options for the disposal of biodegradable food waste. However, the relatively high content of oil in food waste inhibits the conversion efficiency of anaerobic digestion because of the accumulation of long-chain fatty acids (LCFAs). In this study, activated anaerobic sludge was acclimated to accommodate high-oil conditions. The methane yield of high-oil food waste digested by the acclimated sludge increased by 24.9% compared to that digested by the raw sludge. To determine the internal changes in the acclimated sludge, the shifts in the microbial communities during the acclimation period were investigated via high-throughput sequencing (HTS) based on the 16 S rRNA gene. The results indicated that Bacteroidetes, Firmicutes, Chloroflexi and Proteobacteria were the dominant bacteria at the phylum level. The acclimation time allows some functional bacterial taxa to proliferate, such as Clostridium and Longilinea, which are able to degrade LCFAs and turn them into small organic molecules that also have nutrient value for other bacteria. Among the archaeal communities, the hydrogenotrophic methanogen Methanobacterium nearly supplanted the acetotrophic methanogen Methanosaeta. The time profiles of volatile fatty acids (VFAs) and pH during this period provided additional evidence for the success of the acclimation. This study demonstrated the effectiveness of acclimation and the dynamic of microbial communities, which further contributed to the management and resource utilization of high-oil food waste.

Methylmercury (MeHg) is a bioaccumulating neurotoxin mainly produced by anaerobic microorganisms, with methanogen being one of the important methylators. A critical aspect for understanding the mechanism for microbial mercury (Hg) methylation is the origin of the methyl group. However, the origin of methyl group in methanogen-mediated Hg methylation remains unclear. This study aims to identify the source of methyl group for MeHg synthesis in methanogens. Our study revealed that Hg methylation in Methanospirillum hungatei JF-1 is closely related to methanogenesis process, according to the results of proteomic study and substrate limitation study. Next, we proved that nearly all methyl group in MeHg derives from the Wolfe cycle in this species, rather than the previously demonstrated acetyl-coenzyme A pathway, based on the results of 13C labeling study. We then proposed the Wolfe cycle-dependent Hg methylation mechanism in this species. Further genome analyses and 13C labeling experiments indicated that the involvement of the Wolfe cycle in Hg methylation is probably a universal feature among Hg-methylating methanogens. These findings reveal a unique Hg methylation mechanism in methanogens. Our study broadens the carbon substrates and controlling factors for MeHg synthesis in the environment, which can inform the prediction of MeHg production potential and remediation strategies for MeHg contamination.

Coastal wetlands are among the most productive and dynamic ecosystems globally, contributing significantly to atmospheric methane (CH4) emissions. The widespread conversion of these wetlands into aquaculture ponds degrades these ecosystems, yet its effects on CH4 production and associated microbial mechanisms are not well understood. This study aimed to assess the impact of land conversion on CH4 production potential, total and active soil organic C (SOC) content, and microbial communities. We conducted a comparative study on three brackish marshes and adjacent aquaculture ponds in southeastern China. Compared to costal marshes, aquaculture ponds exhibited significantly (P < 0.05) lower CH4 production potential (0.05 vs. 0.02 μg kg-1 h-1), SOC (17.64 vs. 6.97 g kg-1), total nitrogen (TN) content (1.62 vs. 1.24 g kg-1) and carbon/nitrogen (C/N) ratio (10.85 vs. 5.66). CH4 production potential in aquaculture ponds was influenced by both microbial and abiotic factors. Specifically, the relative abundance of Methanosarcina slightly decreased in aquaculture ponds, while the potential for CH4 production declined with lower SOC contents and C/N ratio. Overall, our findings demonstrate that converting natural coastal marshes into aquaculture ponds reduces CH4 production by altering key soil properties and the structure and diversity of methanogenic archaea communities. These results provide empirical evidence to enhance global carbon models, improving predictions of carbon feedback from wetland land conversion in the context of climate change.

Methanogenic communities of hypersaline microbial mats of Guerrero Negro, Baja California Sur, Mexico, have been recognized to be dominated by methylotrophic methanogens. However, recent studies of environmental samples have evidenced the presence of hydrogenotrophic and methyl-reducing methanogenic members, although at low relative abundances. Physical and geochemical conditions that stimulate the development of these groups in hypersaline environments, remains elusive. Thus, in this study the taxonomic diversity of methanogenic archaea of two sites of Exportadora de Sal S.A was assessed by mcrA gene high throughput sequencing from microcosm experiments with different substrates (both competitive and non-competitive). Results confirmed the dominance of the order Methanosarcinales in all treatments, but an increase in the abundance of Methanomassiliiccocales was also observed, mainly in the treatment without substrate addition. Moreover, incubations supplemented with hydrogen and carbon dioxide, as well as the mixture of hydrogen, carbon dioxide and trimethylamine, managed to stimulate the richness and abundance of other than Methanosarcinales methanogenic archaea. Several OTUs that were not assigned to known methanogens resulted phylogenetically distributed into at least nine orders. Environmental samples revealed a wide diversity of methanogenic archaea of low relative abundance that had not been previously reported for this environment, suggesting that the importance and diversity of methanogens in hypersaline ecosystems may have been overlooked. This work also provided insights into how different taxonomic groups responded to the evaluated incubation conditions.

Anaerobic digestion provides a solution for the treatment of vegetable waste water (VWW), but there are currently limited targeted treatment methods available. Building upon previous studies, this research investigated the effects of polyacrylamide-modified magnetic micro-particles (MMP) on anaerobic digestion (AD) of VWW. Three variations of these particles were created by grafting anionic, cationic, and non-ionic polyacrylamide (PAM) onto the MMPs' surfaces, resulting in aPAM-MMP, cPAM-MMP, and nPAM-MMP, respectively. In AD experiments, the addition of aPAM-MMP notably enhanced the degradation of chemical oxygen demand (COD) in VWW. COD decreased to 1290 mg/L in the reactor with aPAM-MMP by day 12 and remained low, while the other reactors had COD concentrations of 4137.5, 5510, and 3010 mg/L on the same day, decreasing thereafter. This modification also improved the production and utilization of hydrogen gas and volatile fatty acids (VFAs), along with the conversion of methane. When tested for bioaffinity using fluorescent GFP-E.coli bacteria, the aPAM-MMP, cPAM-MMP, and nPAM-MMP demonstrated increases in fluorescence intensity by 51.66%, 36.13%, and 37.02%, respectively, compared to unmodified MMP when attached with GFP-E.coli. Further analyses of microbial community revealed that the reactor with aPAM-MMP had the highest microbial richness and enriched bacteria capable of organic matter degradation, such as Bacteroidota, Synergistota, Chloroflexi, Halobacterota phyla, and Parabacteroides, Muribaculaceae, and Azotobacter genera. In conclusion, our experiment verifies that APAM-MMP promotes anaerobic treatment of VWW and provides a novel reference point for enhancing VWW degradation.

Extreme environments, such as highly saline ecosystems, are characterised by a limited presence of microbial communities capable of tolerating and thriving under these conditions. To better understand the limits of life and its chemical and microbiological drivers, highly saline and brine groundwaters of Na-Cl and Na-Ca-Cl types with notably diverse SO4 contents were sampled in water intakes and springs from sedimentary aquifers located in the Outer Carpathians and the Carpathian Foredeep basin and its basement in Poland. Chemical and microbiological methods were used to identify the composition of groundwaters, determine microbial diversity, and indicate processes controlling their distribution using multivariate statistical analyses. DNA sequencing targeting V3-V4 and V4-V5 gene regions revealed a predominance of Proteobacteriota, Methanobacteria, Methanomicrobia, and Nanoarchaea in most of the water samples, irrespective of their geological context. Despite the sample-size constraint, redundancy analysis employing a compositional approach to hydrochemical predictors identified Cl/SO4 and Cl/HCO3 ratios, and specific electrical conductivity, as key gradients shaping microbial communities, depending on the analysed gene regions. Analysis of functional groups revealed that methanogenesis, sulphate oxidation and reduction, and the nitrogen cycle define and distinguish the halotolerant communities in the samples. These communities are characterised by an inverse relationship between methanogens and sulphur-cycling microorganisms.

Anaerobic Membrane Bioreactor (AnMBR) are employed for solid-liquid separation in wastewater treatment, enhancing process efficiency of digestion systems treating digestate. However, membrane fouling remains a primary challenge. This study operated a pilot-scale AnMBR (P-AnMBR) to treat high-concentration organic digestate, investigating system performance and fouling mechanisms. P-AnMBR operation reduced acid-producing bacteria and increased methane-producing bacteria on the membrane, preventing acid accumulation and ensuring stable operation. The P-AnMBR effectively removed COD and VFA, achieving removal rates of 82.3 % and 92.0 %, respectively. Higher retention of organic nitrogen and lower retention of ammonia nitrogen were observed. The membrane fouling consisted of organic substances (20.3 %), predominantly polysaccharides, and inorganic substances (79.7 %), primarily Mg ions (10.1 %) and Ca ions (4.5 %). To reduce the increased transmembrane pressure (TMP) caused by fouling (a 10.6-fold increase in filtration resistance), backwash frequency experiment was conducted. It revealed a 30-min backwash frequency minimized membrane flux decline, facilitating recovery to higher flux levels. The water produced amounted to 70.3 m³ over 52 days. The research provided theoretical guidance and practical support for engineering applications, offering practical insights for scaling up P-AnMBR.

The global climate change mainly caused by fossil fuels combustion promotes that zero-carbon hydrogen production through eco-friendly methods has attracted attention in recent years. This investigation explored the biohydrogen production by co-fermentation of corn straw (CS) and excess sludge (ES), as well as comprehensively analyzed the internal mechanism. The results showed that the optimal ratio of CS to ES was 9:1 (TS) with the biohydrogen yield of 101.8 mL/g VS, which was higher than that from the mono-fermentation of CS by 1.0-fold. The pattern of volatile fatty acids (VFAs) indicated that the acetate was the most preponderant by-product in all fermentation systems during the biohydrogen production process, and its yield was improved by adding appropriate dosage of ES. In addition, the content of soluble COD (SCOD) was reduced as increasing ES, while concentration of NH4+-N showed an opposite tendency. Microbial community analysis revealed that the microbial composition in different samples showed a significant divergence. Trichococcus was the most dominant bacterial genus in the optimal ratio of 9:1 (CS/ES) fermentation system and its abundance was as high as 41.8%. The functional genes prediction found that the dominant metabolic genes and hydrogen-producing related genes had not been significantly increased in co-fermentation system (CS/ES = 9:1) compared to that in the mono-fermentation of CS, implying that enhancement of biohydrogen production by adding ES mainly relied on balancing nutrients and adjusting microbial community in this study. Further redundancy analysis (RDA) confirmed that biohydrogen yield was closely correlated with the enrichment of Trichococcus.

Two records of Earth history capture the evolution of life and its co-evolving ecosystems with interpretable fidelity: the geobiological and geochemical traces preserved in rocks and the evolutionary histories captured within genomes. The earliest vestiges of life are recognized mostly in isotopic fingerprints of specific microbial metabolisms, whereas fossils and organic biomarkers become important later. Molecular biology provides lineages that can be overlayed on geologic and geochemical records of evolving life. All these data lie within a framework of biospheric evolution that is primarily characterized by the transition from an oxygen-poor to an oxygen-rich world. In this Review, we explore the history of microbial life on Earth and the degree to which it shaped, and was shaped by, fundamental transitions in the chemical properties of the oceans, continents and atmosphere. We examine the diversity and evolution of early metabolic processes, their couplings with biogeochemical cycles and their links to the oxygenation of the early biosphere. We discuss the distinction between the beginnings of metabolisms and their subsequent proliferation and their capacity to shape surface environments on a planetary scale. The evolution of microbial life and its ecological impacts directly mirror the Earth's chemical and physical evolution through cause-and-effect relationships.