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Keywords = methane enzyme inhibitor

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26 pages, 11907 KB  
Review
Managing Anti-Nutritional Factors in Plant-Based Feeds: Implications for Herbivore Nutrition and Production
by Mingxia Han, Xiaoyu Liu, Yi Guo, Qingyu Xu, Lin Wei, Jinjin Wei, Muhammad Zahoor Khan, Changfa Wang and Zhenwei Zhang
Metabolites 2026, 16(7), 456; https://doi.org/10.3390/metabo16070456 - 29 Jun 2026
Viewed by 473
Abstract
Anti-nutritional factors (ANFs) in terrestrial plant feeds constrain efficient herbivore production, an issue intensified by rising feed costs and growing demand for animal products. Unlike previous reviews that focus on single ANFs or feed types, this review provides an integrated, cross-species framework linking [...] Read more.
Anti-nutritional factors (ANFs) in terrestrial plant feeds constrain efficient herbivore production, an issue intensified by rising feed costs and growing demand for animal products. Unlike previous reviews that focus on single ANFs or feed types, this review provides an integrated, cross-species framework linking ANF chemistry, rumen microbial interactions, and mitigation strategies. It examines major ANF classes—tannins, phytates, saponins, oxalates, protease inhibitors, lectins, glucosinolates, and gossypol—and their distribution and biochemical modes of action. Mechanistic pathways are grouped into digestive effects (reduced palatability and enzyme inhibition), microbial effects (altered rumen microbiota and fermentation), metabolic effects (impaired absorption), and mineral interactions (nutrient complexation and chelation). Species-specific responses are evaluated, emphasizing the partial detoxification capacity of the rumen microbiome and the dose-dependent nature of ANF effects. Mitigation strategies—physical, chemical, microbial, enzymatic, probiotic, and genetic—are critically assessed for efficacy, scalability, and sustainability. Emerging metabolomic and metagenomic evidence shows that certain ANFs confer functional benefits at controlled doses; for example, tannins improve nitrogen retention, saponins reduce methane, and phytic acid scavenges free radicals. This synthesis supports strategic management rather than complete elimination, informing safe and sustainable use of terrestrial feeds under evolving food-security and environmental challenges. Full article
(This article belongs to the Special Issue Metabolic Responses to Feed and Nutrition in Livestock)
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21 pages, 2198 KB  
Review
Recent Advances and Prospects in Methane Production from Anaerobic Digestion: Process Intensification, Additives, and Biogas Upgrading
by Bonface O. Manono and Felix Lamech Mogambi Ming’ate
Methane 2026, 5(2), 13; https://doi.org/10.3390/methane5020013 - 15 Apr 2026
Cited by 1 | Viewed by 1638
Abstract
Anaerobic digestion (AD) plays an important role in the circular bioeconomy by converting organic waste into renewable methane and nutrient-rich fertilizer. However, consistent, high-quality biomethane production is hindered by four main factors: hydrolysis limitations, fluctuating feedstock quality, microbial instability, and the high cost/energy [...] Read more.
Anaerobic digestion (AD) plays an important role in the circular bioeconomy by converting organic waste into renewable methane and nutrient-rich fertilizer. However, consistent, high-quality biomethane production is hindered by four main factors: hydrolysis limitations, fluctuating feedstock quality, microbial instability, and the high cost/energy demand of purification. This review explores three key areas that improve biomethane production: (i) process intensification (pretreatments and advanced reactors), (ii) microbial regulation through additives, and (iii) biogas upgrading for pipeline use. Anaerobic digestion can be greatly improved by combining thermal or hybrid pretreatments, staged digestion, high-solids technology, and electrochemical systems. These methods speed up hydrolysis and help the system handle higher amounts of organic material more effectively. However, actual performance benefits depend on specific substrate characteristics, heat integration, and control complexity. Optimizing the C:N ratio, buffering capacity, and trace-element supplementation, while simultaneously diluting toxic inhibitors, makes co-digestion an effective and adaptable approach to enhancing anaerobic digestion processes. Additives like carbon, iron nanoparticles, enzymes, and buffers can optimize digestion, but their performance is highly dependent on dosage and substrate. Additionally, they lack validation in long-term, industrial-scale applications. Conventional physicochemical techniques continue to be standard for generating high-quality biomethane, but biological methanation and microalgal systems are playing a growing role in integrating Power-to-Gas technology and using CO2 efficiently. Critical research needs to focus on four areas: (1) standardized reporting metrics, (2) AI-enabled monitoring and control, (3) coupled techno-economic and life-cycle analysis (TEA-LCA), and (4) long-term pilot or full-scale validation. Overall, comprehensive optimization of the entire flow is more effective than improving isolated parts. Full article
(This article belongs to the Special Issue Innovations in Methane Production from Anaerobic Digestion)
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30 pages, 4775 KB  
Article
Uncovering Major Structural and Functional Features of Methyl-Coenzyme M Reductase (MCR) from Methanobrevibacter ruminantium in Complex with Two Substrates
by Han-Ha Chai, Woncheoul Park and Dajeong Lim
Int. J. Mol. Sci. 2026, 27(2), 995; https://doi.org/10.3390/ijms27020995 - 19 Jan 2026
Cited by 1 | Viewed by 685
Abstract
Structural insights into methyl-coenzyme M reductase from Methanobrevibacter ruminantium (M. ruminantium) has implications for methane mitigation strategies. Methanogenesis in ruminants is a major contributor to global greenhouse gas emissions, primarily driven by the rumen archaeon M. ruminantium. Central to this [...] Read more.
Structural insights into methyl-coenzyme M reductase from Methanobrevibacter ruminantium (M. ruminantium) has implications for methane mitigation strategies. Methanogenesis in ruminants is a major contributor to global greenhouse gas emissions, primarily driven by the rumen archaeon M. ruminantium. Central to this process is methyl-coenzyme M reductase (Mcr), an enzyme that catalyzes the final step of methane production. Despite its significance as a chemogenetic target for methane mitigation, the high-resolution structure of M. ruminantium Mcr has remained elusive. Here, we employed homology modeling and CDOCKER simulations within the CHARMM force field to elucidate the structural and functional features of the M. ruminantium Mcr/ligand complexes. We characterized two distinct states: the reduced Mcroxi-silent state bound to HS-CoM and CoB-SH, and the oxidized Mcrsilent state bound to the heterodisulfide CoM-S-S-CoB. Alanine-scanning mutagenesis identified 71 and 62 key residues per active site for each state, respectively, revealing the fundamental determinants of structural stability and substrate selectivity on the Ni-F430 cofactor. Furthermore, structure-based pharmacophore modeling defined essential features (AAADDNNN and AAADDNN) that drive ligand binding. These findings provide a high-resolution molecular framework for the rational design of specific Mcr inhibitors, offering a robust starting point for developing broad-spectrum strategies to suppress enteric methane emissions. Full article
(This article belongs to the Special Issue 25th Anniversary of IJMS: Updates and Advances in Macromolecules)
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15 pages, 710 KB  
Review
Insights into the Feed Additive Inhibitor and Alternative Hydrogen Acceptor Interactions: A Future Direction for Enhanced Methanogenesis Inhibition in Ruminants
by Ibrahim Ahmad, Richard P. Rawnsley, John P. Bowman, Rohan Borojevic and Apeh A. Omede
Fermentation 2025, 11(12), 680; https://doi.org/10.3390/fermentation11120680 - 5 Dec 2025
Cited by 3 | Viewed by 1594
Abstract
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 [...] Read more.
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
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17 pages, 4335 KB  
Article
Development of 18F/11C-Labeled Pyrrolo-Pyridine/Pyrimidine LRRK2 Selective PET Radioligands
by Sangram Nag, Vladimir Stepanov, Akihiro Takano, Ryosuke Arakawa, Marie Svedberg, Nahid Amini, Garrick Paul Smith, Gitte Mikkelsen, Thomas Jensen, Lassina Badolo, Morten Hentzer, Kenneth Vielsted Christensen, Benny Bang-Andersen, Andrea Varrone and Christer Halldin
Pharmaceuticals 2025, 18(12), 1790; https://doi.org/10.3390/ph18121790 - 25 Nov 2025
Cited by 2 | Viewed by 918
Abstract
Background/Objectives: Leucine-rich repeat kinase 2 (LRRK2) is an enzyme implicated in Parkinson’s disease (PD) and a potential therapeutic target. LRRK2 PET radioligands could therefore function as imaging biomarkers for PD and as tools to measure enzyme occupancy of novel therapeutic candidates. This [...] Read more.
Background/Objectives: Leucine-rich repeat kinase 2 (LRRK2) is an enzyme implicated in Parkinson’s disease (PD) and a potential therapeutic target. LRRK2 PET radioligands could therefore function as imaging biomarkers for PD and as tools to measure enzyme occupancy of novel therapeutic candidates. This study aimed at developing novel radioligands for imaging using positron emission tomography (PET). Specific objectives were to synthesize fluorine-18-labeled pyrrolopyridine 1 ([18F]1), pyrrolo-pyrimidine 2 ([18F]2), as well as carbon-11-labeled pyrrolo-pyrimidine 3 ([11C]3), and examine their binding specificity, using in vitro autoradiography (ARG) and in vivo positron emission tomography (PET) imaging in non-human primates (NHPs). Methods: Radiolabeling was achieved either by classical one-step fluorine-18 nucleophilic substitution reaction or by methylation using carbon-11 methane. [18F]1 and [18F]2 were tested in NHP and human whole-hemisphere ARG experiments. PET imaging was performed in cynomolgus monkeys. Radiometabolites were measured in monkey plasma using gradient HPLC. Results: The results demonstrated successful radiolabeling of all three ligands. In ARG studies, both [18F]1 and [18F]2 displayed binding in brain slices from NHP and human samples. The binding of [18F]1 was blocked by cold Compound 1 and structurally distinct Compound 3, but not by the structurally distinct LRRK2 inhibitor PFE-360. On the other hand, the binding of [18F]2 was blocked by PFE-360 in certain regions of the brain, indicating some level of specific binding to LRRK2. All three ligands showed relevant brain uptake (>3%ID), with highest uptake being observed for [18F]1, particularly in the thalamus. In contrast, brain uptake of [18F]2 and [11C]3 was evenly distributed across all brain regions. No blocking effect of [18F]1 was observed after pretreatment with the structurally distinct LRRK2 inhibitors PFE-360 (0.5 mg/kg, iv) and GEN-7915 (40 mg/kg). Conclusions: PET imaging indicated a low in vivo specific binding of the radioligands in the cynomolgus monkey brain, suggesting that the radioligands are not suitable for LRRK2 imaging in vivo with PET. This study emphasizes the challenges in the development of PET radioligands for imaging LRRK2 and the need for additional work to achieve this goal. Full article
(This article belongs to the Section Radiopharmaceutical Sciences)
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14 pages, 1939 KB  
Article
A Meta-Analysis of 3-Nitrooxypropanol Dietary Supplementation on Growth Performance, Ruminal Fermentation, and Enteric Methane Emissions of Beef Cattle
by José Felipe Orzuna-Orzuna, Juan Eduardo Godina-Rodríguez, Jonathan Raúl Garay-Martínez, Lorenzo Danilo Granados-Rivera, Jorge Alonso Maldonado-Jáquez and Alejandro Lara-Bueno
Fermentation 2024, 10(6), 273; https://doi.org/10.3390/fermentation10060273 - 23 May 2024
Cited by 14 | Viewed by 8534
Abstract
This study aimed to evaluate the effects of dietary supplementation with 3-nitrooxypropanol (3-NOP) on growth performance, ruminal fermentation, and enteric methane emissions of beef cattle using a meta-analytic approach. The final meta-analysis database included results from 15 scientific articles. The response variables were [...] Read more.
This study aimed to evaluate the effects of dietary supplementation with 3-nitrooxypropanol (3-NOP) on growth performance, ruminal fermentation, and enteric methane emissions of beef cattle using a meta-analytic approach. The final meta-analysis database included results from 15 scientific articles. The response variables were analyzed through random effects models, where the results were reported as weighted mean differences (WMD) between the treatments without 3-NOP and those supplemented with 3-NOP. The dietary inclusion of 3-NOP decreased (p < 0.001) dry matter intake but did not affect (p > 0.05) average daily gain and increased (p < 0.05) feed efficiency. In the rumen, 3-NOP supplementation increased (p < 0.01) the pH and ruminal concentration of propionate, butyrate, valerate, isobutyrate, and isovalerate. In contrast, dietary supplementation with 3-NOP decreased (p < 0.001) the rumen concentration of ammonia nitrogen, total volatile fatty acids, acetate, and the acetate/propionate ratio. Furthermore, daily methane (CH4) emission, CH4 yield, and CH4 emission as a percentage of gross energy ingested decreased (p < 0.001) in response to 3-NOP dietary supplementation. In conclusion, dietary supplementation with 3-nitrooxypropanol can be used as a nutritional strategy to improve feed efficiency and ruminal fermentation in beef cattle and, at the same time, reduce enteric methane emissions. Full article
(This article belongs to the Special Issue Recent Advances in Rumen Fermentation Efficiency, 2nd Edition)
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29 pages, 7110 KB  
Systematic Review
A Review of Potential Feed Additives Intended for Carbon Footprint Reduction through Methane Abatement in Dairy Cattle
by Ian Hodge, Patrick Quille and Shane O’Connell
Animals 2024, 14(4), 568; https://doi.org/10.3390/ani14040568 - 8 Feb 2024
Cited by 28 | Viewed by 11888
Abstract
Eight rumen additives were chosen for an enteric methane-mitigating comparison study including garlic oil (GO), nitrate, Ascophyllum nodosum (AN), Asparagopsis (ASP), Lactobacillus plantarum (LAB), chitosan (CHI), essential oils (EOs) and 3-nitrooxypropanol (3-NOP). Dose-dependent analysis was carried out on selected feed additives using a [...] Read more.
Eight rumen additives were chosen for an enteric methane-mitigating comparison study including garlic oil (GO), nitrate, Ascophyllum nodosum (AN), Asparagopsis (ASP), Lactobacillus plantarum (LAB), chitosan (CHI), essential oils (EOs) and 3-nitrooxypropanol (3-NOP). Dose-dependent analysis was carried out on selected feed additives using a meta-analysis approach to determine effectiveness in live subjects or potential efficacy in live animal trials with particular attention given to enteric gas, volatile fatty acid concentrations, and rumen microbial counts. All meta-analysis involving additives GO, nitrates, LAB, CHI, EOs, and 3-NOP revealed a reduction in methane production, while individual studies for AN and ASP displayed ruminal bacterial community improvement and a reduction in enteric CH4. Rumen protozoal depression was observed with GO and AN supplementation as well as an increase in propionate production with GO, LAB, ASP, CHI, and 3-NOP rumen fluid inoculation. GO, AN, ASP, and LAB demonstrated mechanisms in vitro as feed additives to improve rumen function and act as enteric methane mitigators. Enzyme inhibitor 3-NOP displays the greatest in vivo CH4 mitigating capabilities compared to essential oil commercial products. Furthermore, this meta-analysis study revealed that in vitro studies in general displayed a greater level of methane mitigation with these compounds than was seen in vivo, emphasising the importance of in vivo trials for final verification of use. While in vitro gas production systems predict in vivo methane production and fermentation trends with reasonable accuracy, it is necessary to confirm feed additive rumen influence in vivo before practical application. Full article
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14 pages, 798 KB  
Review
Potential Effects of Delphinidin-3-O-Sambubioside and Cyanidin-3-O-Sambubioside of Hibiscus sabdariffa L. on Ruminant Meat and Milk Quality
by Rosalba Lazalde-Cruz, Luis Alberto Miranda-Romero, Deli Nazmín Tirado-González, María Isabel Carrillo-Díaz, Sergio Ernesto Medina-Cuéllar, Germán David Mendoza-Martínez, Alejandro Lara-Bueno, Gustavo Tirado-Estrada and Abdelfattah Z. M. Salem
Animals 2021, 11(10), 2827; https://doi.org/10.3390/ani11102827 - 28 Sep 2021
Cited by 18 | Viewed by 6311
Abstract
The objective was to review the potential effects of adding anthocyanin delphinidin-3-O-sambubioside (DOS) and cyanidin-3-O-sambubioside (COS) of HS in animal diets. One hundred and four scientific articles published before 2021 in clinics, pharmacology, nutrition, and animal production were included. [...] Read more.
The objective was to review the potential effects of adding anthocyanin delphinidin-3-O-sambubioside (DOS) and cyanidin-3-O-sambubioside (COS) of HS in animal diets. One hundred and four scientific articles published before 2021 in clinics, pharmacology, nutrition, and animal production were included. The grains/concentrate, metabolic exigency, and caloric stress contribute to increasing the reactive oxygen species (ROS). COS and DOS have antioxidant, antibacterial, antiviral, and anthelmintic activities. In the rumen, anthocyanin might obtain interactions and/or synergisms with substrates, microorganisms, and enzymes which could affect the fiber degradability and decrease potential methane (CH4) emissions; since anthocyanin interferes with ruminal fatty acids biohydrogenation (BH), they can increase the n-3 and n-6 polyunsaturated fatty acids (PUFA), linoleic acid (LA), and conjugated linoleic acid (CLA) in milk and meat, as well as improving their quality. Anthocyanins reduce plasma oxidation and can be deposited in milk and meat, increasing antioxidant activities. Therefore, the reduction of the oxidation of fats and proteins improves shelf-life. Although studies in ruminants are required, COS and DOS act as inhibitors of the angiotensin-converting enzyme (ACEi) and rennin expression, regulating the homeostatic control and possibly the milk yield and body weight. By-products of HS contain polyphenols as calyces with positive effects on the average daily gain and fat meat quality. Full article
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18 pages, 2469 KB  
Article
Isoprene Oxidation by the Gram-Negative Model bacterium Variovorax sp. WS11
by Robin A. Dawson, Nasmille L. Larke-Mejía, Andrew T. Crombie, Muhammad Farhan Ul Haque and J. Colin Murrell
Microorganisms 2020, 8(3), 349; https://doi.org/10.3390/microorganisms8030349 - 29 Feb 2020
Cited by 25 | Viewed by 5349
Abstract
Plant-produced isoprene (2-methyl-1,3-butadiene) represents a significant portion of global volatile organic compound production, equaled only by methane. A metabolic pathway for the degradation of isoprene was first described for the Gram-positive bacterium Rhodococcus sp. AD45, and an alternative model organism has yet to [...] Read more.
Plant-produced isoprene (2-methyl-1,3-butadiene) represents a significant portion of global volatile organic compound production, equaled only by methane. A metabolic pathway for the degradation of isoprene was first described for the Gram-positive bacterium Rhodococcus sp. AD45, and an alternative model organism has yet to be characterised. Here, we report the characterisation of a novel Gram-negative isoprene-degrading bacterium, Variovorax sp. WS11. Isoprene metabolism in this bacterium involves a plasmid-encoded iso metabolic gene cluster which differs from that found in Rhodococcus sp. AD45 in terms of organisation and regulation. Expression of iso metabolic genes is significantly upregulated by both isoprene and epoxyisoprene. The enzyme responsible for the initial oxidation of isoprene, isoprene monooxygenase, oxidises a wide range of alkene substrates in a manner which is strongly influenced by the presence of alkyl side-chains and differs from other well-characterised soluble diiron monooxygenases according to its response to alkyne inhibitors. This study presents Variovorax sp. WS11 as both a comparative and contrasting model organism for the study of isoprene metabolism in bacteria, aiding our understanding of the conservation of this biochemical pathway across diverse ecological niches. Full article
(This article belongs to the Special Issue Microbial Cycling of Atmospheric Trace Gases)
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