Search Results (89)

The urbanization-driven surge in kitchen waste necessitates optimized dry anaerobic digestion (DAD; total solids > 15%). Despite its valorization potential, this technology requires efficiency improvements due to mass transfer constraints. This study evaluated TS effects (15%, 20%, or 25%) on methane production. The TS = 20% system achieved peak cumulative methane yield (405.73 ± 11.71 mL/gVS), exceeding TS = 15% (348.09 ± 12.19 mL/gVS) and TS = 25% (293.08 ± 3.55 mL/gVS). This optimization was attributable to synergistic maintenance of metabolic equilibrium through autonomous pH recovery, rapid VFAs degradation, and enhanced TAN tolerance. Conversely, TS = 25% exhibited impaired mass transfer efficiency under high solids, causing VFAs accumulation, ammonia toxicity, and progressive pH decline to 7.5, indicating system destabilization. Organic degradation analysis confirmed superior conversion efficiency in TS = 20% through dynamic SPS–SPN equilibrium. Microbial analysis revealed enhanced metabolic efficiency via synergistic interactions between acetoclastic and hydrogenotrophic methanogens in TS = 20%. This research provides technical parameters for optimizing methane production in kitchen waste DAD systems. Full article

The quality of strong-flavor baijiu largely depends on the physicochemical properties and prokaryotic microbial activities of pit mud. However, the impact of the iron-bearing minerals in pit mud on its prokaryotic microbial communities remains unknown. This study examined the differences in the prokaryotic communities between 2-year, 40-year, and 100-year pit mud and yellow soil (the raw material for pit mud), as well as the impacts of environmental factors, particularly iron-bearing minerals, on the structure and diversity of these prokaryotic communities. The results indicated that there were significant differences in the composition of prokaryotic microorganisms between yellow soil and pit mud. As the fermentation pit aged, the relative abundance of dominant fermentation bacteria (including Petrimonas, Syntrophomonas, Clostridium, etc.) and hydrogenotrophic methanogens in the pit mud increased. The relative abundance of Lactobacillus in the 2-year pit mud was low (0.33%). Under laboratory conditions, goethite (a typical crystalline iron mineral, denoted as Fe_c) reduced the physiological and metabolic activity of Lacticaseibacillus paracasei JN01 in a concentration-dependent manner. The results of the physicochemical analysis showed that the contents of total iron (TFe) and Fe_c significantly decreased, while the contents of Fe(II) and amorphous iron (hydr)oxides (Fe_o) significantly increased with an increasing fermentation pit age. TFe and Fe_c were significantly negatively correlated with both the Chao1 and Shannon indexes and functional microorganisms such as Clostridium_sensu_stricto_12, Sedimentibacter, and hydrogenotrophic methanogens. The current results contribute to our understanding of the aging process of pit mud from the perspective of the interaction between iron-bearing minerals and prokaryotic microorganisms. Full article

The unambiguous detection of biosignatures in exoplanet atmospheres is a primary objective for astrobiologists and exoplanet astronomers. The primary methodology is the observation of combinations of gases considered unlikely to coexist in an atmosphere or individual gases considered to be highly biogenic. Earth-like examples of the former include CH₄ and O₃, and the latter includes dimethyl sulfide (DMS). To improve the plausibility of the detection of life, I argue that the isotope ratios of key atmospheric species are needed. The C isotope ratios of CO₂ and CH₄ are especially valuable. On Earth, thermogenesis and volcanism result in a substantial difference in δ¹³C between atmospheric CH₄ and CO₂ of ~−25‰. This difference could have changed significantly, perhaps as large as −95‰ after the evolution of hydrogenotrophic methanogens. In contrast, nitrogen fixation by nitrogenase results in a relatively small difference in δ¹⁵N between N₂ and NH₃. Isotopic biosignatures on ancient Earth and rocky exoplanets likely coexist with much larger photochemical signatures. Extreme δ¹⁵N enrichment in HCN may be due to photochemical self-shielding in N₂, a purely abiotic process. Spin-forbidden photolysis of CO₂ produces CO with δ¹³C < −200‰, as has been observed in the Venus mesosphere. Self-shielding in SO₂ may generate detectable ³⁴S enrichment in SO in atmospheres similar to that of WASP-39b. Sufficiently precise isotope ratio measurements of these and related gases in terrestrial-type exoplanet atmospheres will require instruments with significantly higher spectral resolutions and light-collecting areas than those currently available. Full article

Traditional anaerobic digestion (AD) technology continues to have severe limitations in terms of complicated substrate degradation efficiency and methane production. This study optimizes the AD system using corn straw and cattle manure as substrates by introducing an exogenous N-Hexanoyl-L-Homoserine lactone (C6-HSL) signaling molecule in concert with an applied external voltage of 0.8 V, systematically investigating its impact on methanogenic performance and microbial community dynamics. The results show that the combined regulation significantly increased methane production (by 29.74%) and substrate utilization rate (by 74.73%) while preventing acid inhibition and ammonia nitrogen inhibition. Mechanistic analysis revealed that the external voltage enhanced the system’s electrocatalytic activity, while the C6-HSL signaling molecule further facilitated the electron transfer efficiency of the biofilm on the electrode. The combined regulation notably enriched hydrogenotrophic methanogens (with Methanobacterium predominating on the cathode and Methanobrevibacter in the digestate), establishing a stable metabolic cooperative network on both the electrode and in the digestate, optimizing the hydrogenotrophic methanogenesis pathway, and enhancing the synergistic effects among microbial communities and system robustness. This study uncovers the synergistic enhancement mechanism of C6-HSL and external voltage, providing new technological pathways and theoretical support for the efficient conversion of low-quality biomass resources and the production of clean energy. Full article

Anaerobic digestion (AD) is a well-established technology for the sustainable conversion of agricultural organic by-products and waste into bioenergy. Temperature is crucial for optimizing methane production through inocula preservation and reactor start-up in AD. The preservation of inocula induced by temperature has rarely been assessed from an engineering perspective. There has also been limited exploration of the influence of high-to-moderate temperature transition on the initiation of AD. This study employed continuous mesophilic AD reactors with potential engineering applications to conduct revival tests. These tests evaluated the methane production activity of sludge stored at different temperatures and investigated the impact of high-temperature initiation on mesophilic AD. Additionally, we elucidated the correlation between these assessments and microbial diversity as well as composition. The results indicated that bacterial diversity was higher in the inoculum stored at 35 °C compared to 15 °C, ensuring a stable start-up operation of the mesophilic AD. The richness of the bacteria and diversity of the archaea remained stable during the transition from high to mesophilic temperatures. This was conducive to enhancing methanogenic activity of mesophilic AD initiated at 55 °C. The continuously operated AD system showed significant differences in microbial composition compared to its inoculum. Increased abundance of Coriobacteriaceae and Prevotellaceae led to propionate and butyrate accumulation, respectively, reducing AD operational capacity. Methanogenic archaea were less diverse in AD initiated with low-temperature preserved inoculum compared to that with a medium temperature. Streptococcaceae induced by high temperarure could promote AD stability. Hydrogenotrophic methanogens had a competitive advantage in mesophilic AD due to their prior exposure to high-temperature initiation, possibly influenced by Thermotogaceae. Full article

Paddy fields are the main agricultural source of greenhouse gas methane (CH₄) emissions. To enhance rice yield, various fertilization practices have been employed in rice paddies. However, the key microbial and abiotic factors driving CH₄ emissions under different fertilization practices in paddy fields remain largely uncharted. This study conducted field experiments in a traditional double-cropping rice area in South China, utilizing five different fertilization practices to investigate the key factors influencing CH₄ emissions. High-throughput sequencing and PICRUSt2 functional prediction were employed to investigate the contributions of soil physicochemical properties, CH₄-metabolizing microorganisms (methanogens and methanotrophs), and key genes (mcrA and pmoA) on CH₄ emissions. The results showed that CH₄ emission fluxes exhibited seasonal variations, with consistent patterns of change observed across all treatments for both early- and late-season rice. Compared to the no-fertilization (NF) treatment, cumulative CH₄ emissions were lower in early-season rice with green manure (GM) and straw returning (SR) treatments, as well as in late-season rice with GM treatment, while rice yields were maintained at higher levels. High-throughput sequencing analysis revealed that potential methanogens were primarily distributed among four orders: Methanobacteriales, Methanocellales, Methanomicrobiales, and Methanosarcinales. Furthermore, there was a significant positive correlation between the relative abundance of the CH₄-related key gene mcrA and these microorganisms. Functional analysis indicated that these potential methanogens primarily produce methane through the acetoclastic and hydrogenotrophic pathways. Aerobic CH₄-oxidizing bacteria, predominantly from the genus Methylocystis, were detected in all the treatments, while the CH₄ anaerobic-oxidizing archaea ANME-1b was only detected in chemical fertilization (CF) and cow manure (CM) treatments. Our random forest analysis revealed that the relative abundance of two methanogens (Methanocellales and Methanosarcinales) and two environmental factors (pH and DOC) had significant impacts on the cumulative CH₄ emissions. The variance decomposition analysis highlighted the CH₄-metabolizing microorganisms explained 50% of the variance in the cumulative CH₄ emissions, suggesting that they are the key microbial factors driving CH₄ emissions. These findings provide guidance for the development of rational measures to reduce CH₄ emissions in paddy fields. Full article

The first discovery of methanogens led to the formation of a new domain of life known as Archaea. The Archaea domain exhibits properties vastly different from previously known Bacteria and Eucarya domains. However, for a certain multi-step process, a syntrophic relationship between organisms from all domains is needed. This process is called methanogenesis and is defined as the biological production of methane. Different methanogenic pathways prevail depending on substrate availability and the employed order of methanogenic Archaea. Most methanogens reduce carbon dioxide to methane with hydrogen through a hydrogenotrophic pathway. For hydrogen activation, a group of enzymes called hydrogenases is required. Regardless of the methanogenic pathway, electrons are carried between microorganisms by hydrogen. Naturally occurring processes, such as methanogenesis, can be engineered for industrial use. With the growth and emergence of new industries, the amount of produced industrial waste is an ever-growing environmental problem. For successful wastewater remediation, a syntrophic correlation between various microorganisms is needed. The composition of microorganisms depends on wastewater type, organic loading rates, anaerobic reactor design, pH, and temperature. The last step of anaerobic wastewater treatment is production of biomethane by methanogenesis, which is thought to be a cost-effective means of energy production for this renewable biogas. Full article

Ammonium inhibition is a key limiting factor for anaerobic digestion when using chicken manure as the main substrate, especially in a digestion system with biogas slurry reflux. Air stripping is usually used as a recycled biogas slurry treatment. In this study, we carried out the anaerobic co-digestion of chicken manure and corn straw. The fermentation performance was investigated with and without air stripping at different biogas slurry reflux ratios and with an increasing organic loading rate. The results show that air stripping enhanced biogas production, system stability, and volatile solid removal efficiency via the mitigation control of ammonium inhibition. The total ammonium nitrogen in the digesters with air stripping was 20.24–46.40% lower than in those without air stripping. The highest specific biogas production and volatile solid removal efficiency values were obtained in the digesters at an organic loading rate of 3.3 g volatile solid (VS)/(L·d) and a reflux ratio of 75% with air stripping, reaching 480.43 mL/gVS_add and 63.36%, respectively. Moreover, air stripping also improved the organic loading rate and reflux ratio. Stable operation was achieved at an organic loading rate of 5.3 gVS/(L·d) and a reflux ratio of 75%, with specific biogas production of 392.35 mL/gVS_add and a volatile solid removal efficiency of 50.33%. The fermentation performance deteriorated when the organic loading rate was increased to 8.0 gVS/(L·d) at a reflux ratio of 75%, even when air stripping was conducted, indicating that a slighter lower reflux ratio (50%) could be more feasible at a higher organic loading rate (8.0 gVS/(L·d). Additionally, the methanogen community structure varied according to the use of air stripping, with a shift in the methanogenic pathway from hydrogenotrophic to acetoclastic methanogens. Overall, our findings support the adoption of air stripping for ammonium mitigation in anaerobic digestion with biogas slurry reflux. Full article

Reservoirs are a hotspot for methane emissions, a potent greenhouse gas. However, the microbial basis for methane production in the Qinghai Plateau reservoirs remains unclear. To explore the characteristics of methanogenic communities in reservoir sediments on the northeastern Qinghai Plateau, sediment samples were collected from 18 reservoirs in the Yellow River basin during May 2023 (dry season) and August 2023 (wet season). High-throughput sequencing technology was employed to analyze the community composition, diversity, and co-occurrence network of methanogens. Furthermore, FAPROTAX and Mantel analysis were used to assess the metabolic functions of methanogens and their influencing factors. The results showed that (1) the predominant genera of methanogens were Methanobacterium (28.87%) and Methanosarcina (21.67%). Hydrogenotrophic methanogenesis was the main pathway in the sediments. (2) Significant spatiotemporal differences were observed in the diversity of methanogenic communities (p < 0.05). The composition and diversity of these communities were found to be significantly influenced by temperature, pH, altitude, organic carbon, and total nitrogen (p < 0.05). (3) Methanosarcina, Methanobacterium, and Methanospirillum play crucial roles in maintaining the stability of methanogenic community networks. The co-occurrence network nodes are predominantly positively correlated (99.82%). These results provide data for further studies on carbon cycling in the Qinghai Plateau reservoirs. Full article

Biogas upgrading via biomethanation has been extensively studied recently, but the influence of organic loading rate on process performance remains to be fully understood. This is particularly significant because both organic loading rate and hydrogen injection can lead to volatile fatty acid accumulation during anaerobic digestion. This study investigated the impact of a wide range of organic loading rates (from 1.25 to 3.25 g VS/L/d) on hydrogen consumption rates, organic acid accumulation, and microbial communities during in situ biomethanation. It also provided kinetics data and metabolite production data for different control reactors, including anaerobic digestion, ex situ biomethanation, and endogenous control reactors. Hydrogen was injected into parallel batch reactors using digestate from a semi-continuous lab-scale reactor subjected to increasing organic loading rates (1.25–3.25 g VS/L/d) as an inoculum. The inoculum was well adapted to each tested organic loading rate. The batch experiments were replicated following a 12 h hydrogen starvation period to assess the stability of hydrogen consumption rates. High organic loading rate values resulted in increased hydrogen consumption rates, peaking at 68 mg COD/L/h at an organic loading rate of 3.25 g VS/L/d (maximum value tested), with no significant organic acid accumulation despite the high hydrogen partial pressures. The hydrogen consumption rates were maintained after the starvation period. Furthermore, the addition of an organic substrate did not impact the hydrogen consumption rate (i.e., the in situ and ex situ rates were similar). A higher organic loading rate resulted in higher relative abundances of hydrogenotrophic methanogens (i.e., Methanospirillum sp.). This study highlights that increasing the organic loading rate can accelerate the rate of hydrogen consumption during in situ biomethanation, consequently reducing both capital and operational costs. Full article

This study investigated the relationship between water potential (Ψ) and the cation-induced inhibition of methane production in anaerobic digesters. The Ψ around methanogens was manipulated using polyethylene glycol (PEG) in a batch anaerobic reactor, ranging from −0.92 to −5.10 MPa. The ultimate methane potential (B_u) decreased significantly from 0.293 to 0.002 Nm³ kg⁻¹-VS_added as Ψ decreased. When Ψ lowered from −0.92 MPa to −1.48 MPa, the community distribution of acetoclastic Methanosarcina decreased from 59.62% to 40.44%, while those of hydrogenotrophic Methanoculleus and Methanobacterium increased from 17.70% and 1.30% to 36.30% and 18.07%, respectively. These results mirrored changes observed in methanogenic communities affected by cation inhibition with KCl. Our findings strongly indicate that the inhibitory effect of cations on methane production may stem more from the water stress induced by cations than from their direct toxic effects. This study highlights the importance of considering Ψ dynamics in understanding cation-mediated inhibition in anaerobic digesters, providing insights into optimizing microbial processes for enhanced methane production from organic substrates. Full article

This study examined the impact of sodium propionate concentration (0–40 g/L) on the methanogenic archaea in an inoculum which was cultured in basal nutrient medium, exploring its mechanisms and nonlinear stress intensity. The results indicated that at low concentrations, propionate-maintained homeostasis of the anaerobic digestion (AD) system and enriched Methanosaeta. However, when the concentration exceeded 16 g/L, the stability of the AD system was disrupted. The methanogenic pathway shifted towards a predominantly hydrogenotrophic pathway, resulting in a significant increase in methane yield. Below concentrations of 28 g/L, the AD system gradually enhanced its ability to utilize propionate in an orderly manner. At concentrations of 24–28 g/L, genera (e.g., Advenella and Methanosarcina) were enriched to adapt to the high-VFA environment. This was accompanied by a significant upregulation of genes related to the methylotrophic and hydrogenotrophic pathways, effectively mitigating propionate inhibition and enhancing methanogenesis. Conversely, excess concentrations (>30 g/L) suppressed methanogenesis-related genes and led to methane production arrest despite activating specialized propionate-metabolizing bacteria such as genus Pelotomaculum schinkii. As such, an increase in the stress intensity of propionate promotes a change in the metabolic pathways of methanogens and increases methane production; however, excessive sodium propionate was not conducive to maintaining the steady state of the system. Full article

Biochar has attracted increasing attention as an additive for enhancing the performance of anaerobic digestion (AD), but the effect of biochar on microbial regulatory mechanisms in enhancing AD performance is unclear. To investigate how biochar modulates the process of AD, different inoculum sources including cellulose–peptone–swine inoculum (CPI) and swine manure inoculum (SMI) were designed to determine the effect of biochar on the performance and microbial communities of anaerobic digestion of the feedstock concentration from 1 to 6%. The results showed that the methane yields of CPI seeds were higher 20.3–38.7% than those of SMI seeds without a biochar addition, whereas the biochar addition reduced 5.3 and 23.1% of the corresponding methane yield of CPI and SMI, respectively. The biochar enhances the accumulation of volatile fatty acids (VFAs) and weakens the potential ammonia inhibition by adsorption, and it can improve the degradation rate of organic content of soluble COD for different inoculum sources. Microbial community analyses showed that the biochar addition could facilitate the growth of Bacteroidetes and Clostridiales, and it enriched the relative abundance of hydrogenotrophic methanogens Methanobrevibacter and Methanobacterium. Overall, although the modulation of biochar possessed different effects on the anaerobic digestion performance, it contributed to the stability and degradation efficiency of the digestion system. The recycling implication of biochar is critical to realizing a low-carbon and renewable treatment system for organic wastes. Full article

As methane fermentation is inhibited by ammonia derived from organic waste, anaerobic microbial communities tolerant to enriched wastewater with high concentrations of ammonia and salt must be obtained for methane fermentation. Therefore, acclimation cultures were prepared in bottles for 60–80 weeks with artificial wastewater medium added every 2 weeks, using three types of sludge from wastewater treatment plants in food factories. These cultures were maintained without substantially decreasing methanogenesis and gradually increasing NH₄-N and salt concentrations to 5 and 34 g/L, respectively, via the accumulation of ammonia and salt through anaerobic digestion and direct addition. The culture did not show the severe inhibition of methanogenesis or the accumulation of volatile fatty acids (VFAs) such as acetic and propionic acids. The analysis of bacterial consortia in the acclimated sludge based on the 16S rRNA sequence showed that hydrogenotrophic methanogenic bacteria of the genus Methanoculleus were dominant among archaea, whereas bacteria from the orders Clostridiales and Bacteroidales were dominant among eubacteria. Further, VFA-assimilating bacteria, including synthetic acetate-oxidizing bacteria coupled with hydrogenotrophic Methanoculleus to convert methane from acetate, were present to prevent the excessive accumulation of VFAs in the acclimation culture. The proposed acclimation process can enhance the anaerobic digestion of wastewater for methane production. Full article

This study investigates the impacts of bismuth and tin on the production of CH₄ and volatile fatty acids in a microbial electrosynthesis cell with a continuous CO₂ supply. First, the impact of several transition metal ions (Ni²⁺, Fe²⁺, Cu²⁺, Sn²⁺, Mn²⁺, MoO₄²⁻, and Bi³⁺) on hydrogenotrophic and acetoclastic methanogenic microbial activity was evaluated in a series of batch bottle tests incubated with anaerobic sludge and a pre-defined concentration of dissolved transition metals. While Cu is considered a promising catalyst for the electrocatalytic conversion of CO₂ to short chain fatty acids such as acetate, its presence as a Cu²⁺ ion was demonstrated to significantly inhibit the microbial production of CH₄ and acetate. At the same time, CH₄ production increased in the presence of Bi³⁺ (0.1 g L⁻¹) and remained unchanged at the same concentration of Sn²⁺. Since Sn is of interest due to its catalytic properties in the electrochemical CO₂ conversion, Bi and Sn were added to the cathode compartment of a laboratory-scale microbial electrosynthesis cell (MESC) to achieve an initial concentration of 0.1 g L⁻¹. While an initial increase in CH₄ (and acetate for Sn²⁺) production was observed after the first injection of the metal ions, after the second injection, CH₄ production declined. Acetate accumulation was indicative of the reduced activity of acetoclastic methanogens, likely due to the high partial pressure of H₂. The modification of a carbon-felt electrode by the electrodeposition of Sn metal on its surface prior to cathode inoculation with anaerobic sludge showed a doubling of CH₄ production in the MESC and a lower concentration of acetate, while the electrodeposition of Bi resulted in a decreased CH₄ production. Full article