Search Results (85)

Reducing enteric methane emissions from ruminants has emerged as a critical environmental priority in the face of global climate change, given the substantial contribution of methane to agricultural greenhouse gas outputs. This study evaluated the potential of spearmint essential oil (SEO) to reduce methane production and enhance energy utilization efficiency using an in vitro rumen fermentation system. The experiment comprised a control (CON, no additive), three SEO doses (L-SEO: 100 mg/L; M-SEO: 200 mg/L; H-SEO: 400 mg/L), and a commercial essential oil blend (AGL: 150 mg/L). Results indicated that M-SEO and H-SEO significantly reduced methane production at 24 h from 58.11 mL/g DM in CON to 47.93 and 46.58 mL/g DM, respectively (p < 0.001), corresponding to reductions of 17.5% and 19.8%. Furthermore, M-SEO increased total volatile fatty acid concentration from 48.41 to 58.10 mmol/L and elevated the molar proportion of propionate, while significantly enhancing microbial crude protein production (p < 0.001). Microbial community analysis revealed that M-SEO increased bacterial alpha-diversity (Shannon index) (p = 0.001) and significantly enriched specific functional guilds, particularly the propionate-producing genus Succiniclasticum and the butyrate-producing genus Butyrivibrio. Interestingly, the abundance of dominant methanogens (Methanobrevibacter) was not reduced, suggesting a metabolic inhibition mechanism rather than a biocidal effect. Functional prediction analysis further supported this, indicating a downregulation of pathways associated with methanogenesis, including key enzymes such as methyl-coenzyme M reductase. In conclusion, SEO supplementation at 200 mg/L effectively reduced methane production by redirecting metabolic hydrogen toward propionate formation, without affecting overall fermentation. Therefore, the current study indicated that SEO could serve as a sustainable feed additive for mitigating enteric methane emissions in ruminants. Full article

Anaerobic digestion (AD) is a proven and promising technology for recovering energy from biowastes, such as pig slurry (PS) from the fattening/finishing phase. The mechanisms of AD are widely studied, and nowadays, it is of the utmost importance to investigate strategies that give end-users the confidence to choose this technology and to adapt it to their reality, promoting the energy transition and circular economy. This study investigated how collection and storage period affect PS samples, and how hydraulic retention time (HRT) (15 versus 20 days) influences AD performance and stability. Seasonality was the primary factor influencing feedstock characteristics. Samples presented no significant differences during the storage period. A 20-day HRT led to higher digestate pH, total ammonia nitrogen (TAN), and free ammonia nitrogen (FAN) concentrations, which can cause process instability and methanogenesis inhibition. However, 20-day HRT led to a specific methane production that was 7% higher and to a methane quality (expressed in % v/v CH₄) that was 6% higher than 15-day HRT. Overall, methane quality, digestate pH, TAN, and FAN values may be considered key points that need to be monitored to prevent the AD system from being compromised. Nevertheless, these results provide the operational freedom to choose either HRT, allowing reduced reactor volume and investment. Full article

Strategies to suppress methanogenesis must preserve the functional integrity of the rumen microbial ecosystem. Essential oils (EOs) have emerged as promising modulators of rumen microbial function, though their responses vary widely with chemical structure and inclusion level. This study evaluated the efficacy of selected EOs using detailed in vitro fermentation assays. Nine EOs—cinnamon, lavender, garlic (GAR), lemongrass (LEG), peppermint (PPM), eucalyptus, coriander, oregano, and ginger (GIN)—were evaluated for their effects on rumen fermentation and methane (CH₄) production using a 24 h in vitro batch culture system. Eight EOs were tested at two doses (Low and High) specific to each EO, while GIN was evaluated at a single dose. All treatments were incubated in a rumen fluid–buffer mix (1:1 for fermentation parameters and 1:4 for gas and CH₄ measurements) with a 55:45 forage-to-concentrate substrate (pH 6.9). Overall treatment effects were significant for all measured fermentation parameters (p < 0.01). Most treatments reduced total gas production, CH₄ emissions, and CH₄/total gas ratios compared with the control (p < 0.05), although several responses were dose-dependent or directly divergent. Essential oils showed clear, composition-dependent responses: non-terpenoid EOs produced the strongest but also the most variable antimethanogenic effects, with GAR, particularly at the lower dose, consistently achieving the greatest CH₄ inhibition while maintaining a favorable fermentation pattern. Conversely, terpenoid-based EOs induced moderate, dose-responsive CH₄ reductions with minimal effects on overall fermentation. At the higher dose, PPM suppressed CH₄ without altering major volatile fatty acid (VFA) patterns aside from increases in valerate and branched-chain VFA, whereas LEG reduced CH₄ only when accompanied by marked fermentation depression. Monensin validated its role as an effective positive control. Overall, GAR, characterized by sulfur-based bioactives, emerged as the most effective candidate for CH₄ mitigation under the tested in vitro conditions, highlighting the importance of chemical composition and inclusion level in determining efficacy and reinforcing the need for in vivo validation. Full article

High organic loading is known to destabilize anaerobic digestion (AD). This study compared bioaugmentation and pH adjustment under increasing organic loading rate (OLR: 2.0, 4.0 and 6.0 gVS L⁻¹ d⁻¹), focusing on the responses of microbial structure, metabolic pathways, and energy metabolism. Results demonstrated that bioaugmentation maintained stable methane production of 400.54 ± 10.08 and 374.15 ± 24.32 mL·g-VS⁻¹ at 4.0 and 6.0 gVS L⁻¹ d⁻¹, respectively, whereas control and pH-adjusted reactors failed at 4.0 gVS L⁻¹ d⁻¹. The acidified system restored methane yield from 86.30 to 382.13 mL·g-VS⁻¹ after bioaugmentation, whereas pH adjustment and feeding cessation were ineffective, failing to produce methane within 25 days. Microbial analysis showed bioaugmentation enriched Methanosarcina, enhanced hydrogenotrophic/methylotrophic methanogenesis, and strengthened syntrophy with syntrophic propionate-oxidizing bacteria (SPOB), reducing volatile fatty acid accumulation via reinforced syntrophic propionate/butyrate oxidation. Upregulation of osmoregulatory (nha, kdp, proP) and energy metabolism genes (eha, mvh, hdr) maintained osmotic balance and energy supply under high load. In contrast, pH adjustment downregulated SPOB and propionate oxidation genes, causing persistent acid inhibition. This study elucidated the distinct regulatory effects of bioaugmentation and pH adjustment on high-load AD systems, providing actionable strategies for both maintaining operational stability in high-load reactors and recovering methanogenesis in acid-inhibited systems. Full article

Enteric methane (CH₄) emissions from ruminants contribute significantly to agricultural greenhouse gases. Anti-methanogenic feed additives (AMFA), such as Asparagopsis spp. and 3-nitrooxypropanol (3-NOP), reduce CH₄ emissions by inhibiting methanogenic enzymes. However, CH₄ inhibition often leads to dihydrogen (H₂) accumulation, which can impact rumen fermentation and decrease dry matter intake (DMI). Recent studies suggest that co-supplementation of CH₄ inhibitors with alternative electron acceptors, such as phloroglucinol, fumaric acid, or acrylic acid, can redirect excess H₂ during methanogenesis inhibition into fermentation products nutritionally beneficial for the host. This review summarizes findings from rumen simulation experiments and in vivo trials that have investigated the effects of combining a CH₄ inhibitor with an alternative H₂ acceptor to achieve effective methanogenesis inhibition. These trials demonstrate variable outcomes depending on additive combinations, inclusion rates, and adaptation periods. The use of phloroglucinol in vivo consistently decreased H₂ emissions and altered fermentation patterns, promoting acetate production, compared with fumaric acid or acrylic acid as alternative electron acceptors. As a proof-of-concept, phloroglucinol shows promise as a co-supplement for reducing CH₄ and H₂ emissions while enhancing volatile fatty acid profiles in vivo. Optimizing microbial pathways for H₂ utilization through targeted co-supplementation and microbial adaptation could enhance the sustainability of CH₄ mitigation strategies using feed additive inhibitors in ruminants. Further research using multi-omics approaches is needed to elucidate the microbial mechanisms underlying the redirection of H₂ toward beneficial fermentation products during enteric methanogenesis inhibition. This knowledge will help guide the formulation of novel co-supplements designed to reduce CH₄ emissions and improve energy efficiency for sustainable livestock production. Full article

This review relates the kinetics of anaerobic digestion (AD) to energy outcomes, including typical ranges of methane yields and volumetric methane productivities (down to hourly g L⁻¹ h⁻¹ scales relevant for industrial plants). It further translates these relationships into practical control principles that support stable, high methane productivity. Evidence spans substrate selection and co-digestion with emphasis on carbon/nitrogen (C/N) balance, pretreatment strategies, and reactor operation, linking process constraints with operating parameters to identify interventions that raise performance while limiting inhibition. Improving substrate accessibility is the primary step: pretreatment and co-digestion shift limitation beyond hydrolysis and allow safe increases in organic loading. Typical mesophilic operation involves hydraulic retention times of about 10–40 days for food waste and 20–60 days for different types of livestock manure and slowly degradable energy crops, with stable performance achieved when the solids retention time (SRT) is maintained longer than the hydraulic retention time (HRT). Stability is further governed by sustaining a low hydrogen partial pressure through hydrogenotrophic methanogenesis. Temperature and pH define practicable operating ranges; meanwhile, mixing should minimise diffusion resistance without damaging biomass structure. Early-warning indicators—volatile fatty acids (VFAs)/alkalinity, the propionate/acetate ratio, specific methanogenic activity, methane (CH₄)% and gas flow—enable timely adjustment of loading, retention, buffering, mixing intensity and micronutrient supply (Ni, Co, Fe, Mo). In practice, robust operation is generally associated with VFA/alkalinity ratios below about 0.3 and CH₄ contents typically in the range of 50–70% (v/v) in biogas. The review consolidates typical feedstock characteristics and biochemical methane potential (BMP) ranges, as well as outlines common reactor types with their advantages and limitations, linking operational choices to energy yield in combined heat and power (CHP) and biomethane pathways. Reported pretreatment effects span approximately 20–100% higher methane yields; for example, 18–37% increases after mechanical size reduction, around 20–30% gains at 120–121 °C for thermal treatments, and in some cases nearly a two-fold increase for more severe thermal or combined methods. Priorities are set for adaptive control, micronutrient management, biomass-retention strategies, and standardised monitoring, providing a coherent route from kinetic understanding to dependable energy performance and explaining how substrate composition, pretreatment, operating parameters, and kinetic constraints jointly determine methane and energy yield, with particular emphasis on early-warning indicators. Full article

Methane (CH₄), originating from ruminants, is a major source of greenhouse gas emissions in the agriculture industry. This study aimed to determine the potential of red seaweed Gracilaria lemaneiformis (G. lemaneiformis) as an anti-methanogenic feed additive for cattle. Three supplementation levels of seaweed (2%, 5%, and 10% of dry matter) were evaluated for their effects on gas production and rumen fermentation characteristics during 48 h in vitro fermentation. The results revealed a significant decrease in total gas production (TGP), CO₂, CH₄, ammonia nitrogen (NH₃-N), and volatile fatty acid (VFA) concentrations, with no differences in pH or dry matter disappearance (DMD). Notably, compared with the control group without seaweed, supplementation with 2% G. lemaneiformis effectively reduces CH₄ emissions by 27.5% (p < 0.05). Supplementation with 2% G. lemaneiformis decreased the abundance of methanogens g_norank_f_Methanomethylophilaceae, responsible for CH₄ generation, and increased the populations of bacteria (Kandleria and Succinivibrio) that compete with methanogens for substrates. Furthermore, upregulating the levels of 13(S)-HOTrE and 9(S)-HOTrE (polyunsaturated fatty acids) could inhibit methanogenic activity. Additionally, lower VFA concentrations will provide less raw materials for methane synthesis, thus further inhibiting methanogenesis. In summary, G. lemaneiformis, as a red seaweed with important economic value, can not only be applied to enhance marine carbon sinks but can also serve as a promising candidate for mitigating biomethane emissions in cattle. Full article

This study evaluated the effects of feedstuffs and additives in dairy cow rations on rumen methane production and nitrate content in groundwater. Two basal rations and their supplements were analyzed in regard to proximate parameters, and an in vitro rumen fermentation system assessed methane release and nitrate levels over 72 h. Supplementing dairy cow rations with Brassica rapa (BR) boosted the ether extract content, while silage produced the highest amount of methane. Rapidly degrading substrates like BR and ground maize produced methane faster, but in smaller amounts, than straw and silage. BR, Opuntia ficus-indica (OFI), and Posidonia oceanica (PO)-supplemented rations had mixed effects; PO reduced the methane yield, while OFI increased methane production rates. BR-supplemented rations had the lowest nitrate levels, making it suitable for anaerobic digestion. The multivariate analysis showed strong correlations between crude protein, dry matter, and ash, while high-nitrate substrates inhibited methane production, supporting the literature on the role of nitrates in reducing methanogenesis. These results emphasize the need to balance nutrient composition and methane mitigation strategies in dairy cow ration formulations. Full article

Leachate management remains a major environmental challenge, especially in rapidly urbanizing cities of developing countries. Traditionally considered toxic and useless, it is a sustainable organic resource with the potential for high-value biochemical production through bioprocessing. This study investigated the characteristics of fresh leachates from three solid waste transfer stations (SWTS) in the Abidjan district, Côte d’Ivoire, and assessed their potential as substrates for medium-chain fatty acid (MCFA) production via microbial chain elongation. The MCFA synthesis was carried out in anaerobic bioreactors operated under methanogenesis inhibition conditions. The leachates from Bingerville, Abobo-Dokui, and Yopougon exhibited acidic and high organic content, particularly volatile fatty acids (VFAs), key precursors for MCFA synthesis. High concentrations of microbial communities associated with chain elongation were observed, including Clostridium (sulphite-reducing), Lactobacillus, Bacillus, and Pseudomonas (greater than 5 log10 CFU/mL). MCFA production ranged from 5 to 10 g/L, mainly C6, C7, and C8, with compositional variation depending on the SWTS. Notably, leachates from higher-income areas demonstrated higher MCFA productivity compared to those from lower-income areas. These findings highlight the potential of fresh SWTS leachates in the Abidjan district for sustainable MCFA production, paving the way for industrial applications. Full article

Livestock methane emissions are a significant source of greenhouse gases. The aim of this study was to investigate the secondary metabolites of different strains of silage quinoa and their impact on methane emissions from livestock farming. In this study, we evaluated the chemical composition, fermentation quality, secondary metabolite content, and in vitro gas production of eight quinoa lines, 093, 137, 231, 238, 565, 666, 770, and 811, grown in saline and alkaline areas of the Yellow River Delta. The results showed that crude protein, EE, and crude ash content ranged from 8.84% to 10.69%, 1.98% to 2.38%, and 17.00% to 23.14%, respectively. The acidic and neutral detergent fiber content of these eight quinoa varieties ranged from 49.31% to 61.91% and 33.29% to 37.31%, respectively. Line 093 had the highest total saponin content, while Line 231 exhibited the highest flavonoid content. Methane yield was significantly and negatively correlated with tannin, saponin, and flavonoid content, whereas carbon dioxide yield showed a positive correlation with saponin and flavonoid content. Among all lines, 770 and 811 demonstrated the lowest methane production, indicating strong in vitro inhibition of methanogenesis. These findings suggest that feeding quinoa silage to ruminants has the potential to reduce greenhouse gas emissions. Full article

Earth’s atmospheric methane (CH₄) concentration has risen more than 162% since pre-industrial levels in the mid-18th century, and about 30% of the rise in global temperatures since the pre-industrial era is due to CH₄ The build-up of methane in the atmosphere in 2020–2022 was the largest since systematic measurements started in 1983, more than double the average yearly growth rate measured over the previous 17 years (15.2 ppb yr⁻¹ vs. 5.71 ppb yr⁻¹, respectively). During 2020, with a growth rate of 14.81 ppb yr⁻¹, the level of atmospheric CH₄ broke the previous record (which was set in 1991), and it was broken again immediately the following year, with an increase of 17.64 ppb yr⁻¹ in 2021. For 2022, the final estimate is 13.25 ppb yr⁻¹, the fourth largest annual growth rate. The most recent explanations for this surge in tropospheric CH₄ include increased emissions from tropical wetlands, more floods, and increased temperatures. For 2020 and part of 2021, a reduction in the oxidative capacity of the atmosphere due to COVID-19 lockdowns was also proposed. Our main hypothesis is that this CH₄ surge in 2020–2021 may also be caused by reduced sulfate emissions, which have been shown to decrease methanotrophy and increase methanogenesis rates in wetlands. Then, for the CH₄ slowdown in 2022–2024, our hypotheses are that the emissions from wetlands remained high, but that there was an even higher increase in the oxidative capacity of the atmosphere due to multiple other parameters that are detailed in this article. This perspective review paper is mainly qualitative; it demonstrates that coupled climate–chemistry models will also need to integrate biochemistry, as the evolution of the atmospheric composition is multifactorial and non-linear. Full article

Volatile fatty acids (VFAs) are inevitable intermediates of biogas production during the anaerobic digestion of organic matter. The excessive accumulation of VFAs leads to a pH drop and the strong inhibition of methanogenesis. On the other hand, VFAs are useful commodities with different applications, and their fermentative production may compete with traditional production methods based on oil derivatives. The fermentation methods have commonalities with the biorefinery concept. The present review considers the methods of VFA fermentative production together with competitive simultaneous biogas and hydrogen production. Methods of the enhanced production of volatile fatty acids are presented, showing the option of integrated processes of product removal and energy production from the obtained biogas. On the basis of the present review, the following conclusion can be drawn. Volatile fatty acids (formic, acetic, propionic, and butyric ones) are useful commodities with various applications. That is why their targeted production with their desired production rate may shift the aims of the anaerobic digestion toward volatile fatty acids instead of biogas release. On the other hand, VFA production combined with biogas release can make the overall process self-consistent, with energy production sufficient to maintain the target processes using biogas for heating the digestor. The maintenance of optimum VFA concentrations can be accomplished by simultaneous VFA removal from the fermentation broth, thus integrating the product recovery with the maintenance of optimum operation conditions in the digester. The substrate preparation and the operating conditions (organic loading rate and hydraulic retention time) are of crucial importance for the successful fermentation process. Full article

This study, conducted on a commercial dairy farm using a robotic milking system in Victoria, Australia, examined the effects of Polygain™ (The Product Makers Australia), a polyphenol-rich sugarcane feed material (PRSFM), on CH₄ emissions in grazing dairy cattle using an inverse-dispersion model (IDM) combined with open-path laser techniques. Thirty lactating Holstein Friesian cows (aged 2–5 years with an average body weight of 663 kg and average daily milk production of 28.9 kg) were divided into two dietary treatment groups of fifteen cows each. Before the measurement, the PRSFM (0.25%) was supplemented for 3 weeks as an adaptation period and mixed with pellet feed for the treatment group. Over the 2-week measurement period, CH₄ production (MP) was 495 ± 12 and 377 ± 12 (mean ± standard error) g CH₄/animal/day for the control and treatment groups, respectively. Methane intensity (MI) was 17.04 and 13.01 g CH₄/animal/kg milk/day in the control and treatment groups, respectively. On average, Polygain supplementation reduced MP and MI by 24%. This potential CH₄ reduction extrapolated across Australia contributes to a 2.63% reduction in national agricultural emissions. This study underscores the potential of Polygain for CH₄ mitigation in dairy cattle. Full article

This study examines the anaerobic digestion (AD) of fruit and vegetable waste (FVW) and digestates to assess the effect of particle size on anaerobic biodegradability (AB) and process rate at different stages of digestion progress. Batch assays were conducted with FVW mixtures and digestates from 5, 10, and 15 days of digestion, using four particle size ranges: Ø1 < 1.8 µm, 1.8 < Ø2 < 500 µm, 500 < Ø3 < 1000 µm, and Ø4 > 1000 µm. While AB and specific methanogenic activity (SMA) showed no significant differences among FVW mixtures, particle size significantly influenced these variables. Methane yields were 298.2 and 309.8 $\mathrm{m}\mathrm{L} \mathrm{C}{\mathrm{H}}_4·{\mathrm{g}}^{-1}\mathrm{V}\mathrm{S}$ for Ø4 and Ø3 particles, exceeding the 186.7 and 161.8 $\mathrm{m}\mathrm{L} \mathrm{C}{\mathrm{H}}_4·{\mathrm{g}}^{-1}\mathrm{V}\mathrm{S}$ obtained for Ø2 and Ø1, respectively. These results indicate that particle size reduction enhanced methane production; however, reducing the particle size below 500 µm inhibits methanogenesis due to intermediate accumulation. Compared to FVW, digestates exhibited a 23% reduction in AB, a 73.9% decrease in SMA, and methane yields of 55.8–294 $\mathrm{m}\mathrm{L} \mathrm{C}{\mathrm{H}}_4·{\mathrm{g}}^{-1}\mathrm{V}\mathrm{S}$. Additionally, the surface-based kinetic constant ($K_{SBK}$) decreased from 0.4523 $\mathrm{g}·\mathrm{d}{\mathrm{m}}^{-2}·{\mathrm{d}}^{-1}$for FVW to 0.0437 $\mathrm{g}·\mathrm{d}{\mathrm{m}}^{-2}·{\mathrm{d}}^{-1}$for digestates. These differences are attributed to the rapid consumption of easily biodegradable fractions within the first 5 days of digestion. 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