Methanogenesis inhibition remodels microbial fermentation and stimulates acetogenesis in ruminants

G Gaofeng Ni (Department of Microbiology, Biomedicine Discovery Institute, Monash University) M Min Wang N Nicola Walker (Animal Nutrition and Health, dsm-firmenich) S Stefan Muetzel (Grasslands Research Centre, AgResearch Limited) O Oliver Schmidt (Department of Arctic and Marine Biology, Faculty of Biosciences, Fisheries and Economics, Universitetet i Tromsø -The Arctic University of Norway) A Andre Fischer R René T. Stemmler (Chemical and Process Sciences, dsm-firmenich) P Pok Man Leung (Department of Microbiology, Biomedicine Discovery Institute, Monash University) X Xiumin Zhang (State Key Laboratory of Forage Breeding-by-Design and Utilization, Institute of Subtropical Agriculture, Chinese Academy of Sciences) Q Qiushuang Li (State Key Laboratory of Forage Breeding-by-Design and Utilization, Institute of Subtropical Agriculture, Chinese Academy of Sciences) S Surbhi Jain (Department of Microbiology, Biomedicine Discovery Institute, Monash University) M Marion Jespersen (Department of Microbiology, Biomedicine Discovery Institute, Monash University) R Rhys Grinter (Department of Microbiology, Biomedicine Discovery Institute, Monash University) S Stephen D. J. Archer (Grasslands Research Centre, AgResearch Limited) D David Pacheco (Grasslands Research Centre, AgResearch Limited) K Katherine Lowe (Grasslands Research Centre, AgResearch Limited) P Phillip B. Pope (Centre for Microbiome Research, School of Biomedical Sciences, Faculty of Health, Queensland University of Technology) V Volker Müller (Department of Molecular Microbiology and Bioenergetics, Institute of Molecular Biosciences, Johann Wolfgang Goethe University) D Dipti W. Pitta (Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania) P Peter H. Janssen (Grasslands Research Centre, AgResearch Limited) M Mick Watson (Data Science and Research, dsm-firmenich) G Graeme T. Attwood (Grasslands Research Centre, AgResearch Limited) E Emiel Ver Loren van Themaat (Data Science and Research, dsm-firmenich) M Maik Kindermann (Animal Nutrition and Health, dsm-firmenich) C Chris Greening

Abstract

Rumen microbiota enable ruminants to grow on fibrous plant materials, but also produce methane, driving 5% of global greenhouse gas emissions and leading to a loss of gross energy content. Methanogenesis inhibitors such as 3-nitrooxypropanol (3-NOP) decrease methane emissions in ruminants when supplemented in feed. Yet we lack a system-wide, species-resolved understanding of how the rumen microbiota remodels following inhibition and how this influences animal production. Here, we conducted a large-scale trial with 51 dairy calves to analyze microbiota responses to 3-NOP, pairing host performance, emissions, and nutritional profiles with genome-resolved metagenomic and metatranscriptomic data. 3-NOP supplementation decreased methane emissions by 62%, modulated short-chain fatty acid and H 2 levels, and did not affect dietary intake or animal performance. We created a rumen microbial genome catalogue (27,884 genomes) that mapped to the meta-omic data at high rates. There was a strong reduction of methanogens and stimulation of reductive acetogens, primarily uncultivated lineages such as “ Candidatus Faecousia.” However, there was a shift in major fermentative communities away from acetate production in response to hydrogen gas accumulation. In vitro incubations recapitulated these results and showed an enrichment of acetate from reductive acetogenesis. Altogether, the divergent responses of the fermentative and hydrogenotrophic communities lead to net hydrogen build-up and limit potential productivity gains from methane reduction. By linking ruminant greenhouse gas emissions and productivity to specific microbial species, this study emphasizes the importance of microbiota-wide analysis for optimizing methane mitigation strategies and identifies promising strategies to simultaneously reduce emissions while increasing animal production.

Article Details

Volume / Issue Vol. 122, Issue 41
Published October 14, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (25)

G

Gaofeng Ni

Department of Microbiology, Biomedicine Discovery Institute, Monash University

M

Min Wang

N

Nicola Walker

Animal Nutrition and Health, dsm-firmenich

S

Stefan Muetzel

Grasslands Research Centre, AgResearch Limited

O

Oliver Schmidt

Department of Arctic and Marine Biology, Faculty of Biosciences, Fisheries and Economics, Universitetet i Tromsø -The Arctic University of Norway

A

Andre Fischer

R

René T. Stemmler

Chemical and Process Sciences, dsm-firmenich

P

Pok Man Leung

Department of Microbiology, Biomedicine Discovery Institute, Monash University

X

Xiumin Zhang

State Key Laboratory of Forage Breeding-by-Design and Utilization, Institute of Subtropical Agriculture, Chinese Academy of Sciences

Q

Qiushuang Li

State Key Laboratory of Forage Breeding-by-Design and Utilization, Institute of Subtropical Agriculture, Chinese Academy of Sciences

S

Surbhi Jain

Department of Microbiology, Biomedicine Discovery Institute, Monash University

M

Marion Jespersen

Department of Microbiology, Biomedicine Discovery Institute, Monash University

R

Rhys Grinter

Department of Microbiology, Biomedicine Discovery Institute, Monash University

S

Stephen D. J. Archer

Grasslands Research Centre, AgResearch Limited

D

David Pacheco

Grasslands Research Centre, AgResearch Limited

K

Katherine Lowe

Grasslands Research Centre, AgResearch Limited

P

Phillip B. Pope

Centre for Microbiome Research, School of Biomedical Sciences, Faculty of Health, Queensland University of Technology

V

Volker Müller

Department of Molecular Microbiology and Bioenergetics, Institute of Molecular Biosciences, Johann Wolfgang Goethe University

D

Dipti W. Pitta

Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania

P

Peter H. Janssen

Grasslands Research Centre, AgResearch Limited

M

Mick Watson

Data Science and Research, dsm-firmenich

G

Graeme T. Attwood

Grasslands Research Centre, AgResearch Limited

E

Emiel Ver Loren van Themaat

Data Science and Research, dsm-firmenich

M

Maik Kindermann

Animal Nutrition and Health, dsm-firmenich

C

Chris Greening