Vaccination of sheep with a methanogen protein provides insight into levels of antibody in saliva needed to target ruminal methanogens
Open accessAbstract
Methane is produced in the rumen of ruminant livestock by methanogens and is a major contributor to agricultural greenhouse gases. Vaccination against ruminal methanogens could reduce methane emissions by inducing antibodies in saliva which enter the rumen and impair ability of methanogens to produce methane. Presently, it is not known if vaccination can induce sufficient amounts of antibody in the saliva to target methanogen populations in the rumen and little is known about how long antibody in the rumen remains active. In the current study, sheep were vaccinated twice at a 3-week interval with a model methanogen antigen, recombinant glycosyl transferase protein (rGT2) formulated with one of four adjuvants: saponin, Montanide ISA61, a chitosan thermogel, or a lipid nanoparticle/cationic liposome adjuvant (n = 6/formulation). A control group of sheep (n = 6) was not vaccinated. The highest antigen-specific IgA and IgG responses in both saliva and serum were observed with Montanide ISA61, which promoted levels of salivary antibodies that were five-fold higher than the second most potent adjuvant, saponin. A rGT2-specific IgG standard was used to determine the level of rGT2-specific IgG in serum and saliva. Vaccination with GT2/Montanide ISA61 produced a peak antibody concentration of 7 × 1016 molecules of antigen-specific IgG per litre of saliva, and it was estimated that in the rumen there would be more than 104 molecules of antigen-specific IgG for each methanogen cell. Both IgG and IgA in saliva were shown to be relatively stable in the rumen. Salivary antibody exposed for 1-2 hours to an in vitro simulated rumen environment retained approximately 50% of antigen-binding activity. Collectively, the results from measuring antibody levels and stablility suggest a vaccination-based mitigation strategy for livestock generated methane is in theory feasible.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Vaccination can induce the host immune system to create antibodies capable of suppressing methanogens, potentially reducing CH4 emissions, particularly in pasture-based systems.
“Vaccination, by inducing the host immune system to create antibodies capable of suppressing methanogens, has the potential to reduce CH4 emissions and is particularly beneficial for pasture-based systems.130”
From Technologies and perspectives for achieving carbon neutrality (2021) · cites this paper as [130]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
R: A Language and Environment for Statistical Computing
R Core Team (2000) · doi:10.32614/r.manuals · Cited by 355,706 · see citing works
Controlling the False Discovery Rate: A Practical and Powerful Approach to Multiple Testing
Benjamini, Y., & Hochberg, Y. (1995) · Journal of the Royal Statistical Society Series B: Statistical Methodology · doi:10.1111/j.2517-6161.1995.tb02031.x · Cited by 109,482 · see citing works
Veterinary Immunology: An Introduction (2000)
(2001) · Journal of Equine Veterinary Science · doi:10.1016/s0737-0806(01)70088-3 · Cited by 914 · see citing works
Stoline, M. R. (1981) · The American Statistician · doi:10.1080/00031305.1981.10479331 · Cited by 280 · see citing works
Cationic lipids activate intracellular signaling pathways
Lonez, C., Vandenbranden, M., & Ruysschaert, J.-M. (2012) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2012.05.009 · Cited by 221 · see citing works
The rate of flow and composition of various salivary secretions in sheep and calves
Kay, R. N. B. (1960) · The Journal of Physiology · doi:10.1113/jphysiol.1960.sp006402 · Cited by 205 · see citing works
Veterinary Immunology: An Introduction
Beveridge, W. I. B. (1993) · Australian Veterinary Journal · doi:10.1111/j.1751-0813.1993.tb00833.x · Cited by 198 · see citing works
Strategies to reduce methane emissions from farmed ruminants grazing on pasture
Buddle, B. M., Denis, M., Attwood, G. T., Altermann, E., Janssen, P. H., Ronimus, R. S., Pinares-Patiño, C. S., Muetzel, S., & Wedlock, D. N. (2010) · The Veterinary Journal · doi:10.1016/j.tvjl.2010.02.019 · Cited by 187 · see citing works
Self-citationReducing methane emissions in sheep by immunization against rumen methanogens
WRIGHT, A. (2004) · Vaccine · doi:10.1016/j.vaccine.2004.03.053 · Cited by 176 · see citing works
Chitosan-based systems for the delivery of vaccine antigens
Arca, H. Ç., Günbeyaz, M., & Şenel, S. (2009) · Expert Review of Vaccines · doi:10.1586/erv.09.47 · Cited by 167 · see citing works
A Vaccine against Rumen Methanogens Can Alter the Composition of Archaeal Populations
Williams, Y. J., Popovski, S., Rea, S. M., Skillman, L. C., Toovey, A. F., Northwood, K. S., & Wright, A. G. (2009) · Applied and Environmental Microbiology · doi:10.1128/aem.02453-08 · Cited by 151 · see citing works
Immunization with Streptococcus bovis protects against lactic acidosis in sheep
Gill, H. S., Shu, Q., & Leng, R. (2000) · Vaccine · doi:10.1016/s0264-410x(00)00017-7 · Cited by 82 · see citing works
In vivo evaluation of chitosan as an adjuvant in subcutaneous vaccine formulations
Scherließ, R., Buske, S., Young, K., Weber, B., Rades, T., & Hook, S. M. (2013) · Vaccine · doi:10.1016/j.vaccine.2013.07.081 · Cited by 82 · see citing works
Self-citationProgress in the development of vaccines against rumen methanogens
Wedlock, D. N., Janssen, P. H., Leahy, S. C., Shu, D., & Buddle, B. M. (2013) · animal · doi:10.1017/s1751731113000682 · Cited by 79 · see citing works
Self-citationSeedorf, H., Kittelmann, S., & Janssen, P. H. (2014) · Applied and Environmental Microbiology · doi:10.1128/aem.03018-14 · Cited by 69 · see citing works
Self-citationThe effect of fleece on core and rumen temperature in sheep
Beatty, D. T., Barnes, A., Fleming, P. A., Taylor, E., & Maloney, S. K. (2008) · Journal of Thermal Biology · doi:10.1016/j.jtherbio.2008.07.002 · Cited by 59 · see citing works
Immunisation against lactic acidosis in cattle
SHU, Q., Gill, H. S., Hennessy, D., Leng, R. A., Bird, S. H., & Rowe, J. B. (1999) · Research in Veterinary Science · doi:10.1053/rvsc.1998.0284 · Cited by 57 · see citing works
Gordon, S., Teichmann, E., Young, K., Finnie, K. S., Rades, T., & Hook, S. M. (2010) · European Journal of Pharmaceutical Sciences · doi:10.1016/j.ejps.2010.07.004 · Cited by 57 · see citing works
Self-citationRumen Volume as a Factor Involved in Individual Sheep Differences
Purser, D., & Moir, R. (1966) · Journal of Animal Science · doi:10.2527/jas1966.252509x · Cited by 44 · see citing works
Williams, Y. J., Rea, S. M., Popovski, S., Pimm, C. L., Williams, A. J., Toovey, A. F., Skillman, L. C., & Wright, A. G. (2007) · British Journal Of Nutrition · doi:10.1017/s0007114507801553 · Cited by 37 · see citing works
Subharat, S., Shu, D., Zheng, T., Buddle, B. M., Janssen, P. H., Luo, D., & Wedlock, D. N. (2015) · Veterinary Immunology and Immunopathology · doi:10.1016/j.vetimm.2015.02.008 · Cited by 36 · see citing works
Self-citationReducing microbial ureolytic activity in the rumen by immunization against urease therein
Zhao, S., Wang, J., Zheng, N., Bu, D., Sun, P., & Yu, Z. (2015) · BMC Veterinary Research · doi:10.1186/s12917-015-0409-6 · Cited by 33 · see citing works
SHU, Q., Gill, H. S., Leng, R. A., & Rowe, J. B. (2000) · The Veterinary Journal · doi:10.1053/tvjl.1999.0400 · Cited by 33 · see citing works
Immunogenicity of Liposomes Containing Lipid Core Peptides and the Adjuvant Quil A
White, K. L., Rades, T., Kearns, P. S., Tóth, I., & Hook, S. M. (2006) · Pharmaceutical Research · doi:10.1007/s11095-006-0272-z · Cited by 27 · see citing works
Self-citationAntibody Response in Sheep Following Immunization with Streptococcus bovis in Different Adjuvants
Shu, Q., Bird, S. H., Gill, H. S., Duan, E., Xu, Y., Hillard, M. A., & Rowe, J. B. (2001) · Veterinary Research Communications · doi:10.1023/a:1026757917968 · Cited by 21 · see citing works
Molecular cloning and physiological effects of brushtail possum interleukin-1β
Wedlock, D. N., Goh, L. P., Parlane, N. A., & Buddle, B. M. (1999) · Veterinary Immunology and Immunopathology · doi:10.1016/s0165-2427(99)00004-5 · Cited by 20 · see citing works
Self-citationShu, Q., Hillard, M. A., Bindon, B. M., Duan, E., Xu, Y., Bird, S. H., Rowe, J. B., Oddy, V. H., & Gill, H. S. (2000) · American Journal of Veterinary Research · doi:10.2460/ajvr.2000.61.839 · Cited by 20 · see citing works
Shu, Q., Bird, S. H., Gill, H. S., & Rowe, J. B. (1999) · FEMS Immunology & Medical Microbiology · doi:10.1111/j.1574-695x.1999.tb01383.x · Cited by 11 · see citing works
Water kinetics in the rumen of two breeds of sheep
Ranilla, M., López, S., Giráidez, F., Valdés, C., & Carro, M. D. (1995) · Annales de Zootechnie · doi:10.1051/animres:199505140 · Cited by 2 · see citing works
Flagged first, then by citations
1 of the 30 references couldn’t be matched to a known paper.
Cited by
Sorted by citations
Technologies and perspectives for achieving carbon neutrality
Wang, F., Harindintwali, J. D., Yuan, Z., Wang, M., Wang, F., Li, S., Yin, Z., Huang, L., Fu, Y., Li, L., et al. (2021) · The Innovation · doi:10.1016/j.xinn.2021.100180 · Cited by 1,330 · see citing works
Beauchemin, K. A., Ungerfeld, E. M., Eckard, R. J., & Wang, M. (2020) · Animal · doi:10.1017/s1751731119003100 · Cited by 619 · see citing works
Invited review: current enteric methane mitigation options
Beauchemin, K. A., Ungerfeld, E. M., Abdalla, A. L., Alvarez, C., Arndt, C., Becquet, P., Benchaar, C., Berndt, A., Mauricio, R. M., McAllister, T. A., et al. (2022) · Journal of Dairy Science · doi:10.3168/jds.2022-22091 · Cited by 324 · see citing works
Mitigation Pathways Compatible with 1.5°C in the Context of Sustainable Development
IPCC (2022) · Cambridge University Press eBooks · doi:10.1017/9781009157940.004 · Cited by 167 · see citing works
Strategies Used to Reduce Methane Emissions from Ruminants: Controversies and Issues
Króliczewska, B., Pecka‐Kiełb, E., & Bujok, J. (2023) · Agriculture · doi:10.3390/agriculture13030602 · Cited by 157 · see citing works
Reisinger, A. R., Clark, H., Cowie, A., Emmet‐Booth, J., Fischer, C. G., Herrero, M., Howden, M. S., & Leahy, S. C. (2021) · Philosophical Transactions of the Royal Society A Mathematical Physical and Engineering Sciences · doi:10.1098/rsta.2020.0452 · Cited by 143 · see citing works
Potential solutions to the major greenhouse-gas issues facing Australasian dairy farming
Eckard, R., & Clark, H. (2018) · Animal Production Science · doi:10.1071/an18574 · Cited by 46 · see citing works
Li, Q., Ma, Z., Huo, J., Zhang, X., Wang, R., Zhang, S., Jiao, J., Dong, X., Janssen, P. H., Ungerfeld, E. M., et al. (2024) · The ISME Journal · doi:10.1093/ismejo/wrad016 · Cited by 45 · see citing works
Self-citationAre Vaccines the Solution for Methane Emissions from Ruminants? A Systematic Review
Baca-González, V., Asensio‐Calavia, P., González-Acosta, S., de la Lastra, J. M. P., & Morales-delaNuez, A. (2020) · Vaccines · doi:10.3390/vaccines8030460 · Cited by 45 · see citing works
Malyugina, S., Holik, S., & Horký, P. (2025) · Frontiers in Animal Science · doi:10.3389/fanim.2025.1610376 · Cited by 33 · see citing works
Goopy, J. P. (2019) · Animal Production Science · doi:10.1071/an18457 · Cited by 31 · see citing works
Mitigating greenhouse gas emissions from New Zealand pasture-based livestock farm systems
Leahy, S. C., Kearney, L., Reisinger, A. R., & Clark, H. (2019) · Journal of New Zealand Grasslands · doi:10.33584/jnzg.2019.81.417 · Cited by 21 · see citing works
Khanum, S., Carbone, V., Gupta, S. K., Yeung, J. H. F., Shu, D., Wilson, T., Parlane, N. A., Altermann, E., Estein, S. M., Janssen, P. H., et al. (2022) · Scientific Reports · doi:10.1038/s41598-022-14545-8 · Cited by 18 · see citing works
Self-citationTailored Nanoparticles With the Potential to Reduce Ruminant Methane Emissions
Altermann, E., Reilly, K., Young, W., Ronimus, R. S., & Muetzel, S. (2022) · Frontiers in Microbiology · doi:10.3389/fmicb.2022.816695 · Cited by 15 · see citing works
The Path to Net-Zero in Dairy Production: Are Pronounced Decreases in Enteric Methane Achievable?
Beauchemin, K. A., Kebreab, E., Cain, M. L., & VandeHaar, M. J. (2024) · Annual Review of Animal Biosciences · doi:10.1146/annurev-animal-010324-113703 · Cited by 13 · see citing works
Vannier, C., Cochrane, T. A., Zawar‐Reza, P., & Bellamy, L. (2022) · Land · doi:10.3390/land11122334 · Cited by 8 · see citing works
Revisiting Environmental Sustainability in Ruminants: A Comprehensive Review
Shang, Y., Ju, T., Kaur, U., Mulim, H. A., Singh, S., Boerman, J. P., & de Oliveira, H. R. (2026) · Agriculture · doi:10.3390/agriculture16020149 · Cited by 7 · see citing works
Khanum, S., Roberts, J., Heathcott, R. W., Bagley, S., Wilson, T., Gupta, S. K., Kirk, M. R., Heiser, A., Janssen, P. H., & Wedlock, D. N. (2022) · Frontiers in Microbiology · doi:10.3389/fmicb.2022.918111 · Cited by 6 · see citing works
Self-citationMorsy, A. S., Soltan, Y. A., Al‐Marzooqi, W., & El‐Zaiat, H. M. (2025) · Sustainability · doi:10.3390/su17146458 · Cited by 5 · see citing works
Multi-omics in vitro study of the salivary modulation of the goat rumen microbiome
Palma‐Hidalgo, J. M., Belanche, A., Jiménez, E., Martín-García, A. I., Newbold, C. J., Denman, S. E., & Yáñez-Ruíz, D. R. (2023) · animal · doi:10.1016/j.animal.2023.100895 · Cited by 5 · see citing works
Meng, F., Wang, Y., Chen, C., Pan, T., Li, J., Xu, Y., Wang, Z., Yao, H., Jiao, X., & Yin, Y. (2025) · Veterinary Research · doi:10.1186/s13567-025-01483-2 · Cited by 4 · see citing works
Ahmad, I., Rawnsley, R., Bowman, J. P., & Omede, A. A. (2025) · Journal of Dairy Science · doi:10.3168/jds.2025-26915 · Cited by 4 · see citing works
Du, X., Liu, X., Mawolo, J. B., Wang, H., Mi, X., Dong, J., Li, Q., & Wen, Y. (2020) · Veterinary Medicine and Science · doi:10.1002/vms3.236 · Cited by 2 · see citing works
Probiotics Against Methanogens and Methanogenesis
Singh, B., Mal, G., Kalra, R. S., & Marotta, F. (2024) · Springer eBooks · doi:10.1007/978-3-031-65455-8_16 · Cited by 1 · see citing works
Liquid Addition Techniques to Enhance Methane Biotrickling Filters at Dairy Barn Concentrations
Pryor, A. M., Gostomski, P. A., & Carere, C. R. (2025) · Clean Technologies · doi:10.3390/cleantechnol8010003 · Cited by 1 · see citing works
Bhandhari, N., Mehla, D., Yadav, S., Joshi, H., Prameela, D. R., Murag, S. B., Bhatnagar, R., & Sharma, M. K. (2026) · Plant Physiology Reports · doi:10.1007/s40502-026-01000-3 · Cited by 1 · see citing works
van Laar, H., Bannink, A., van Gastelen, S., & Šebek, L. B. J. (2023) · doi:10.18174/641132
Wang, M., Li, Q., Ma, Z., Huo, J., Zhang, X., Wang, R., Zhang, S., Jiao, J., Dong, X., Janssen, P. H., et al. (2024) · Research Square · doi:10.21203/rs.3.rs-2831003/v1
Self-citationTemmerman, G. D. (2022) · EDP Sciences eBooks · doi:10.3917/edp.colle.2022.01.0053
Mapping the Spatial Distribution of Socio-economic Facilities in Asaba Delta State
Ojanikele, W. A. (2025) · International Journal of Multidisciplinary Research and Growth Evaluation · doi:10.54660/.ijmrge.2025.6.4.854-862
Shuttleworth, F., & Trees, L. (2025) · In Practice · doi:10.1002/inpr.70007
Yeung, J., Hannaford, R., Wilson, T., Ferguson, T., Khanum, S., McNeilly, T. N., Janssen, P. H., & Wedlock, D. N. (2026) · Frontiers in Immunology · doi:10.3389/fimmu.2026.1797719
Self-citationEn, B., Burke, J., Khan, A., Wedlock, D. N., Janssen, P. H., & Jonker, A. (2026) · New Zealand Veterinary Journal · doi:10.1080/00480169.2026.2695397
Self-citation
- Authors
- Subharat, S., Shu, D., Zheng, T., Buddle, B. M., Kaneko, K., Hook, S., Janssen, P. H., & Wedlock, D. N.
- Affiliations
- AgResearch (NZ)
- University of Otago (NZ)
- Corresponding author
- Subharat, Supatsak
- ORCID
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2016-07-29
- Volume
- 11
- Issue
- 7
- Pages
- e0159861
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC4966943
- OpenAlex
- W2487117742
- Cited by
- 32
- References
- 30
- Citations by year
- 2026: 4 · 2025: 6 · 2024: 4 · 2023: 3 · 2022: 7 · 2021: 2 · 2020: 3 · 2019: 2 · 2018: 1
- FWCI
- 2.05
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- PLoS ONE (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Figshare (submittedVersion, cc-by)
- Funders
- New Zealand Agricultural Greenhouse Gas Research Centre
- Topics
- Life Sciences › Agricultural and Biological Sciences › Agronomy and Crop Science › Ruminant Nutrition and Digestive Physiology
- Physical Sciences › Engineering › Building and Construction › Anaerobic Digestion and Biogas Production
- Life Sciences › Immunology and Microbiology › Microbiology › Microbial infections and disease research
- Keywords
- sheep, salivary antibodies, IgA, IgG, adjuvants, saponins, rumen environment, antibody stability, greenhouse gas emissions
- MeSH terms
- Animals, Antibodies, Antigens, Archaea, Enzyme-Linked Immunosorbent Assay, Female, Immunoglobulin A, Immunoglobulin G, Rumen, Saliva, Vaccination, Archaeal Proteins, Sheep, Domestic
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
