Cited by 496Ruminant Nutrition and Digestive PhysiologyAgriculture Sustainability and Environmental ImpactAnimal Nutrition and Physiology

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An inhibitor persistently decreased enteric methane emission from dairy cows with no negative effect on milk production

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Hristov, Alexander N., Oh, Joonpyo, Giallongo, Fabio, Frederick, Tyler W., Harper, Michael T., Weeks, Holley L., Branco, Antonio F., Moate, Peter J., Deighton, Matthew H., Williams, S. Richard O., Kindermann, Maik, Duval, Stephane·2015·Proceedings of the National Academy of Sciences

DOI 10.1073/pnas.1504124112PMC PMC4553761

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Abstract

A quarter of all anthropogenic methane emissions in the United States are from enteric fermentation, primarily from ruminant livestock. This study was undertaken to test the effect of a methane inhibitor, 3-nitrooxypropanol (3NOP), on enteric methane emission in lactating Holstein cows. An experiment was conducted using 48 cows in a randomized block design with a 2-wk covariate period and a 12-wk data collection period. Feed intake, milk production, and fiber digestibility were not affected by the inhibitor. Milk protein and lactose yields were increased by 3NOP. Rumen methane emission was linearly decreased by 3NOP, averaging about 30% lower than the control. Methane emission per unit of feed dry matter intake or per unit of energy-corrected milk were also about 30% less for the 3NOP-treated cows. On average, the body weight gain of 3NOP-treated cows was 80% greater than control cows during the 12-wk experiment. The experiment demonstrated that the methane inhibitor 3NOP, applied at 40 to 80 mg/kg feed dry matter, decreased methane emissions from high-producing dairy cows by 30% and increased body weight gain without negatively affecting feed intake or milk production and composition. The inhibitory effect persisted over 12 wk of treatment, thus offering an effective methane mitigation practice for the livestock industries.

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Background
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Background
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Assessment of Cleantech Options to Mitigate the Environmental Impact of South African Dairy Farming
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BackgroundMethodInfluential
Effect of canola oil supplementation level on total tract digestion, ruminal fermentation, and methane emissions of cows grazing Urochloa sp. supplemented with a fixed amount of concentrate
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Insights into Effects of Combined Capric and Lauric Acid on Rumen Bacterial Composition
Vadroňová, M., Šťovíček, A., Výborná, A., Tyrolová, Y., Tichá, D., & Joch, M. (2024) · Microorganisms · doi:10.3390/microorganisms12061085 · Cited by 3
Assessing the Impact of the Methane Inhibitors 3-Nitrooxypropanol (3-NOP) and Canola Oil on the Rumen Anaerobic Fungi
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“The targeted nature of 3-NOP is a direct consequence of its highly selective mode of action, acting as a competitive inhibitor of the methyl-coenzyme M reductase (MCR) enzyme, which is ubiquitously and uniquely found in methanogenic archaea [10,75].”
Hiện trạng và kịch bản giảm phát thải khí mêtan từ đường tiêu hóa của hệ thống nuôi bò thịt quảng canh quy mô nông hộ ở Quảng Ngãi
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Duurzaamheids-ethische toetsing van methaanreducerende technische maatregelen : Evaluatie van technische maatregelen ten aanzien van de integrale verduurzaming van de veehouderij
Puente-Rodríguez, D., & Groenestein, C. M. (2019) · doi:10.18174/494713 · Cited by 3
Comparative analysis of macroalgae supplementation on the rumen microbial community: Asparagopsis taxiformis inhibits major ruminal methanogenic, fibrolytic, and volatile fatty acid-producing microbes in vitroPreprint
O’Hara, E., Moote, P. E., Terry, S. A., Beauchemin, K. A., McAllister, T., Abbott, D. W., & Gruninger, R. J. (2022) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2022.09.08.507231 · Cited by 3
Salvianolic acid C inhibits methane emissions in dairy cows by targeting MCR and reshaping the rumen microbial community
Liu, Z., Li, X., Tang, X., He, Y., Nan, X., Wang, H., Guo, Y., & Xiong, B. (2025) · Journal of Animal Science and Biotechnology/Journal of animal science and biotechnology · doi:10.1186/s40104-025-01285-8 · Cited by 3
“3-NOP is an artificially synthesized compound that can inhibit CH 4 emissions, but it also causes a significant increase in H 2 emissions, and the subsequent metabolic fate of increased H 2 remains unclear [25].”
A Physics-Based Framework for Accurate On-Farm Enteric Methane Sensing
Muriel, D. F., & Jaramillo-Botero, A. (2026) · ACS Agricultural Science & Technology · doi:10.1021/acsagscitech.5c00958 · Cited by 2
A meta-analysis establishes bromoform dose and forage-based models to evaluate the antimethanogenic effects of Asparagopsis spp. feed additive
Goloja, C., Povejsil, N., Roque, B. M., Akbay, A., & Honan, M. C. (2026) · BMC Agriculture · doi:10.1186/s44399-025-00030-w · Cited by 2
“This interaction is particularly important, as enteric CH 4 production in ruminants results in a loss of gross energy intake, thereby diminishing the over-all efficiency of nutrient utilization [16].”
Enteric methane emissions and mitigation strategies within pasture-based dairy systems
Kay, J., Garrett, K., Hammond, K. J., Koning, L., Muñoz, C., Martin, C., & Lahart, B. (2026) · JDS Communications · doi:10.3168/jdsc.2025-0980 · Cited by 2
“…baseline methane emissions and the response to mitigation vary with the basal diet, cow intake, stage of lactation, and the mitigation strategy (Hristov et al., 2015; Pickering et al., 2015; Eugène et al., 2021; Beauchemin et al., 2022), and thus should be evaluated under representative…”
3N Agriculture: Growing the greenest tomorrow
Field, D. J., Minasny, B., & McBratney, A. B. (2026) · Agricultural & Environmental Letters · doi:10.1002/ael2.70068 · Cited by 3
“Hristov et al., 2015; Teague et al., 2016).”
Dietary inclusion of Asparagopsis taxiformis significantly reduces methane emissions in dairy cows by mechanistically altering vitamin B12-dependent and other methanogenesis precursor pathways
Lawther, K., Dimonaco, N. J., Donnelly, P., Guinguina, A., Krizsan, S. J., & Huws, S. (2026) · Microbiome · doi:10.1186/s40168-026-02447-0 · Cited by 3
“…CH 4 emissions from ruminants have been achieved through innovative feeding strategies, such as the addition of the chemical 3-nitroxypropanol (3-NOP, Bovaer ® ) to the diet, resulting in approximately 25–30% reductions in CH 4, irrespective of the units used to express CH 4 emissions [1, 22].”
Efecto de la variedad de orujo de uva sobre la producción de gas y metano in vitro
Barraso, C., López-Parra, M. M., & Rodríguez, P. (2021) · Archivos de Zootecnia · doi:10.21071/az.v70i269.5416 · Cited by 2
“El exceso de H2 generado en la conversión de glucosa a AGV y el CO2 producido por la descarboxilación de metabolitos durante la fermentación, son utilizados como sustratos por las arqueas y protozoos, para transformarlos en CH4 (Hook et al. 2010; Alayon et al. 2018).”
Background
Environmental impact of feed additives in cattle production
Kebreab, E. (2023) · American Association of Bovine Practitioners Conference Proceedings · doi:10.21423/aabppro20228607 · Cited by 3
Estrategias de nutrición y alimentación para reducir las emisiones de metano en rumiantes bajo sistemas pastoriles: Una revisión
Peña-Avelino, L. Y., Alva-Pérez, J., Rosales-Martínez, G. N., ́ández-Contreras, S. H., & Ceballos‐Olvera, I. (2024) · Ciencias Veterinarias y Producción Animal · doi:10.29059/cvpa.v1i2.18 · Cited by 3
Lactational performance and enteric methane emissions in dairy cows fed high-oil oats, cold-pressed rapeseed cake, and 3-nitrooxypropanol in a grass silage–based diet
Fant, P., Mantovani, G., Vadroňová, M., Sabetti, M. C., Krizsan, S. J., & Ramin, M. (2025) · Journal of Dairy Science · doi:10.3168/jds.2025-27007 · Cited by 2
Ten promising pathways to GHG emission reduction in the global food system
Pingault, N., & Martius, C. (2023) · Center for International Forestry Research (CIFOR) eBooks · doi:10.17528/cifor-icraf/008836 · Cited by 3
The economics of globally shared goods
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Methane Production in Dairy Cows, Inhibition, Measurement, and Predicting Models
Ramin, M., Chagas, J. C. C., & Krizsan, S. J. (2020) · Springer eBooks · doi:10.1007/978-981-15-4728-7_10 · Cited by 2
Sublime Grasslands: A Story of the Pampas, Prairie, Steppe, and Savannas Where Animals Graze
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Impact of Emerging Agricultural Contaminants on Global Warming
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Assessing the potential of a novel feed additive and an unsaturated fat alone and in combination to lower methane emission from cattle and reduce their contribution to climate change
Smith, M. L. (2017) · Library, Museums and Press - UDSpace (University of Delaware) · doi:10.58088/15qs-z729 · Cited by 2
Metagenomic analysis of marsupial gut microbiomes provides a genetic basis for the low methane economyPreprint
Bowerman, K. L., Lü, Y., McRae, H., Volmer, J. G., Zaugg, J., Pope, P. B., Hugenholtz, P., Greening, C., Morrison, M., Soo, R. M., et al. (2024) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2024.12.04.626884 · Cited by 2
Effect of amino acid supplementation and choline chloride for low protein diet on nitrogen efficiency and methane emission of dairy cows
Shirmohammadi, S., Taghizadeh, A., Hosseinkhani, A., Janmohammadi, H., Pirmohammadi, R., & Valizadeh, H. (2021) · Semina Ciências Agrárias · doi:10.5433/1679-0359.2022v43n1p159 · Cited by 1
“…bacteria, whether by vaccine and or antimicrobial compounds, was assumed to be the main strategy yet (Wedlock, Janssen, Leahy, Shu, & Buddle, 2013; Hristov et al., 2015; Weimar et al., 2017; Henderson, Cook, & Ronimus, 2018) feed or nutritional management could be an alternative policy (Kiggundu…”
Background
Quantifying H2 emissions under different nutritional mitigation strategies and its impact on improving the prediction of enteric methane emissions of ruminants
Mazo, S. B., Ochoa, S. L. P., Noguera, R. R., Bayat, A. R., Benaouda, M., Blondiaux, P., & Eugène, M. (2023) · Livestock Science · doi:10.1016/j.livsci.2023.105342 · Cited by 1
Exploring the Catrina, an autochthonous cattle breed of the Azores, for a comparative analysis of methane emissions with Holstein-Friesian dairy cows
Azevedo, A. R., Lopes, M. S., Borba, A. E. S., Machado, A. D. C., & Mendonça, D. (2024) · Frontiers in Animal Science · doi:10.3389/fanim.2024.1423940 · Cited by 1
Impact of Zootechnical Additive of the Nitrooxypropanol to Minimize Methane Emission in Cattle: A Review
Mshary, G. S., Castillo, C., Bermúdez, J. H., & Muíño, R. (2024) · Diyala Agricultural Sciences Journal · doi:10.52951/dasj.24160203 · Cited by 1
Evaluating the Efficacy of a Novel Multi-Component Feed Additive for Methane Mitigation and Performance Enhancement in Sheep
Rivero, M. J., Khan, A. A., Akpensuen, T. T., Méo-Filho, P., Márquez, S. P., & Jones, A. (2025) · Ruminants · doi:10.3390/ruminants5020017 · Cited by 1
“Compared to additives like 3-NOP, which can achieve greater methane suppression in cattle [23], NuAdvent+ offers a moderate yet promising reduction, particularly suited to forage-based systems.”
Endo-1,3-β-D-glucanase activity influences in vitro ruminal fermentation of diets varying in forage:concentrate
Seeforth, D. P., & Trotta, R. J. (2025) · Journal of Animal Science · doi:10.1093/jas/skaf244 · Cited by 1
Greenhouse gases emission from dairy industries and promising abatement processes and technologies
Udourioh, G. A., Adewumi, C. N., Okolie, J. A., Udourioh, M. G., & Solomon, M. M. (2025) · International Journal of Dairy Technology · doi:10.1111/1471-0307.70040 · Cited by 1
Roles of dandelions alone or in combination with lactic acid bacteria on improving aerobic stability and mitigating in vitro greenhouse gas emissions in whole-plant corn silage
Guo, L., Zhang, M., Dang, H., He, M., Han, M., Zhang, S., Fan, W., Jiang, D., Liu, X., Cui, Y., et al. (2025) · Journal of Integrative Agriculture · doi:10.1016/j.jia.2025.08.005 · Cited by 1
Effect of a Corn Silage-Based Finishing Diet on Growth, Carcass Composition, Meat Quality, Methane Emissions and Carbon Footprint of Crossbred Angus Young Bulls
Soares, D. M., Bernardino, S., Rodrigues, N. R., Gama, I., Almeida, J. M., Teixeira, R. F. M., Santos‐Silva, J., Alves, S. P., Domingos, T., Martin, C., et al. (2025) · Sustainability · doi:10.3390/su17188417 · Cited by 2
Investigating interactions between grass silage– or corn silage–based diets and 3-nitrooxypropanol on fermentation and methane emission dynamics and rumen microbiome in dairy cattle
van Gastelen, S., Dijkstra, J., Alferink, S. J. J., Binnendijk, G. P., Klop, A., van Themaat, E. V. L., Walker, N. D., & Bannink, A. (2025) · Journal of Dairy Science · doi:10.3168/jds.2025-26957 · Cited by 2
A bromoform based Investigational Veterinary Product fed twice daily to lactating dairy cows had no adverse effects on eating, rumination, or locomotion behaviours
Tognelli, R., Alvarez-Hess, P. S., Neachtain, A. S. Ó., Chandra, S., Williams, S. R. O., Jacques, S., Denman, S. E., Eckard, R., & Jacobs, J. L. (2025) · Veterinary and Animal Science · doi:10.1016/j.vas.2025.100539 · Cited by 1
“The use of methane mitigants has been associated with increased hydrogen production (Hristov et al., 2015; Vyas et al., 2018; Roque et al., 2019), elevated lactate, formate and ethanol concentrations (Melgar et al., 2020a), and raised ruminal hydrogen partial pressure (Martinez-Fernandez et al.,…”
Livestock methane emissions in China: Spatiotemporal dynamics and mitigation strategies
Kang, X., Du, M., Du, H., Liu, Q., Zhuang, M., Su, H., Wang, J., Yang, Y., Zhao, X., & Zhu, Q. (2025) · Resources Conservation and Recycling · doi:10.1016/j.resconrec.2025.108695 · Cited by 2
A data fusion approach unveils the effects of 3-nitrooxypropanol on the rumen fluid and milk metabolomes of lactating Holstein dairy cows
Rocchetti, G., Froldi, F., Lapris, M., Moschini, M., Bani, P., Cattaneo, L., & Trevisi, E. (2026) · Journal of Dairy Science · doi:10.3168/jds.2025-26969 · Cited by 2
Deploying anaerobic digestion for animal manure in China by 2030 maximizes short-term methane mitigation benefits
Li, B., Sun, H., Chen, P., Hu, J., Wei, Q., Dong‬, R., & Guo, J. (2026) · Resources Conservation and Recycling · doi:10.1016/j.resconrec.2026.108868 · Cited by 1
Solutions for methane mitigation in Brazilian agriculture: achieving a 28% reduction by 2035
Quintana, G. D. O., Potenza, R. F., & Oliveira, S. L. L. (2026) · Frontiers in Sustainable Food Systems · doi:10.3389/fsufs.2026.1773678 · Cited by 1
Chicory reduces enteric methane emissions and maintains milk yield but decreases milk fat content compared with perennial ryegrass in dairy cows
Cui, Z.-Y., Zom, R. L. G., Strikwold, M., Bannink, A., Pellikaan, W. F., & Dijkstra, J. (2026) · Journal of Dairy Science · doi:10.3168/jds.2025-28034 · Cited by 1
In Vitro screening of methanogenesis inhibitors and dihydrogen sinks for mitigating methane emissions in ruminants
Menni, G., Giudice, F., Battelli, M., Dai, X., Rapetti, L., Galassi, G., Ramin, M., Crovetto, G. M., & Colombini, S. (2026) · Frontiers in Animal Science · doi:10.3389/fanim.2026.1816504 · Cited by 1
Proanthocyanidins inhibit methane emissions by interacting with methyl-coenzyme M reductase and reshaping rumen microbiome function
Liu, Z. H., Guo, Y., Xiao, L., Guo, J., Chen, Y., Wang, H., Nan, X., Zhou, M., Zhang, F., He, Y., et al. (2026) · Microbiome · doi:10.1186/s40168-026-02406-9 · Cited by 1
“3-nitrooxypropanol 3-nitrooxypropanol is a synthetic compound that targets MCR activity and has been shown to reduce CH 4 emissions of dairy cows by more than 30% [13].”
Chemical Characterization and Rumen-Modulating Effects of Pinus sylvestris Essential Oil: In Vitro and In Vivo Study
Pachura, N., Lewandowska, K., Burek, A., Szumny, A., Tabiś, A., Banaszkiewicz, S., Bania, J., & Kupczyński, R. (2026) · Molecules · doi:10.3390/molecules31101769 · Cited by 1
“Measurements were performed on days 0, 3, 7, and 14 under standardized conditions.”
Enteric Methane Emissions from Holstein Cows Grazing Kikuyu Grass in the Colombian High Tropics During Two Seasons
Cruz, L. J. J., Macías, K. N. M., Henao, S. C., Lobo, M. E. A., Cuervo, W., Correa, M. V. G., Cardona, H. J. C., Riomalo, J. E. E., & Mejía, Á. M. G. (2026) · Animals · doi:10.3390/ani16111662 · Cited by 1
“…because it enables direct and continuous measurement of exhaled gases during behavioral states including grazing, rumination, and resting, which are typically underrepresented or not directly determined by other techniques (i.e., GreenFeed system [90,91], SF 6 [92], respiration chamber [93,94]).”

Authors: Hristov, A. N., Oh, J., Giallongo, F., Frederick, T. W., Harper, M. T., Weeks, H. L., Branco, A. F., Moate, P. J., Deighton, M. H., Williams, S. R. O., Kindermann, M., & Duval, S.
Affiliations: Pennsylvania State University (US)
Departamento de Zootecnia, Universidade Estadual de Maringá, PR 87020-900, Brazil;
Department of Economic Development Jobs Transport and Resources (AU)
Animal Nutrition and Health, DSM Nutritional Products, Basel CH-4002, Switzerland;
Research Centre for Animal Nutrition and Health, DSM Nutritional Products France, Saint Louis Cedex 68305, France
ORCID: Hristov, Alexander N.
Oh, Joonpyo
Harper, Michael T.
Moate, Peter J.
Deighton, Matthew H.
Williams, S. Richard O.
Kindermann, Maik
Venue: Proceedings of the National Academy of Sciences
Publisher: National Academy of Sciences
Published: 2015-07-30
Volume: 112
Issue: 34
Pages: 10663-10668
ISSN: 0027-8424 (ISSN-L)
1091-6490
DOI: 10.1073/pnas.1504124112
PMC: PMC4553761
OpenAlex: W2145651012
Cited by: 496
References: 59
Citations by year: 2026: 54 · 2025: 49 · 2024: 58 · 2023: 48 · 2022: 48 · 2021: 53 · 2020: 40 · 2019: 42 · 2018: 35 · 2017: 31 · 2016: 34 · 2015: 4
FWCI: 74.95
Type: journal article
Language: en
Update notices: correction (2015-08-12)
Open access: bronze
doi.org
Locations: Proceedings of the National Academy of Sciences (publishedVersion)
Europe PMC (PubMed Central) (publishedVersion)
PubMed Central (publishedVersion)
PubMed (publishedVersion)
Seoul National University Open Repository (Seoul National University) (submittedVersion)
Topics: Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Ruminant Nutrition and Digestive Physiology
Physical Sciences › Environmental Science › Ecology › Agriculture Sustainability and Environmental Impact
Life Sciences › Agricultural and Biological Sciences › Animal Science and Zoology › Animal Nutrition and Physiology
Keywords: enteric methane, 3-nitrooxypropanol, methane inhibitors, dairy cattle, Holstein cows, methane mitigation, milk production, milk protein, lactose yield, weight gain, ruminant methane emissions, feed intake, fiber digestibility
MeSH terms: Medicago sativa, Animals, Animal Feed, Archaea, Energy Intake, Carbon Dioxide, Cattle, Zea mays, Female, Fermentation, Gases, Hydrogen, Lactation, Methane, Milk, Rumen, Weight Gain, Greenhouse Effect, Dietary Supplements, Propanols

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An inhibitor persistently decreased enteric methane emission from dairy cows with no negative effect on milk production

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