title: Development of a vaccine to mitigate greenhouse gas emissions in agriculture: vaccination of sheep with methanogen fractions induces antibodies that block methane production in vitro.
abstract: OBJECTIVE
  To develop an understanding of the immune responses of ruminants to methanogens, and to provide proof of a concept that harnessing the immune system of ruminants is a potentially viable approach to mitigate greenhouse gas emissions from agriculture.
  
  
  METHODS
  Four subcellular fractions, namely cytoplasmic, two cell-wall preparations, and cell wall-derived proteins were prepared from Methanobrevibacter ruminantium M1. Twenty sheep (10 months of age) were vaccinated with these fractions or with whole cells (n=4 per group). Sheep were re-vaccinated once after 3 weeks, and antibody responses to M. ruminantium M1 antigens in sera and saliva measured using ELISA at 2 weeks after the second vaccination. Antigens recognised by the antisera were visualised using Western blotting. The antisera were tested in vitro for their impact on M. ruminantium M1, measuring the effect on cell growth, methane production, and ability to induce agglutination.
  
  
  RESULTS
  Basal levels (pre-vaccination) of antibodies against M. ruminantium M1 antigens were low. Vaccination with the antigenic fractions induced strong antibody responses in serum. Both IgG and IgA responses to methanogen antigens were detected in saliva following vaccination. Western blot analysis of the antisera indicated reactivity of antibodies, and a wide range of proteins was present in the different methanogen fractions. Antisera against the various fractions agglutinated methanogens in an in-vitro assay. In addition, these antisera decreased the growth of a pure culture of a methanogen and production of methane in vitro.
  
  
  CONCLUSIONS
  Antigens from methanogens are immunogenic in ruminants, and antisera from sheep vaccinated with fractions of methanogens have a significant impact on these organisms, inducing cell agglutination, and decreasing growth of methanogens and production of methane. Only antisera to selected methanogen fractions were able to achieve these effects. The results demonstrate the feasibility of a vaccination strategy to mitigate emission of methane.
authors.affiliation: AgResearch, Hopkirk Research Institute, Grasslands Research Centre, Private Bag 11008, Palmerston North 4442, New Zealand. neil.wedlock@agresearch.co.nz
authors.forename: D N
authors.initials: DN
authors.lastname: Wedlock
authors.forename: G
authors.initials: G
authors.lastname: Pedersen
authors.forename: M
authors.initials: M
authors.lastname: Denis
authors.forename: D
authors.initials: D
authors.lastname: Dey
authors.forename: P H
authors.initials: PH
authors.lastname: Janssen
authors.forename: B M
authors.initials: BM
authors.lastname: Buddle
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D000907:Antibodies, Bacterial / Q000097:blood; D001428:Bacterial Vaccines / Q000276:immunology*; D017752:Greenhouse Effect; D008297:Male; D008697:Methane / Q000276:immunology* / Q000378:metabolism*; D041581:Methanobrevibacter / Q000276:immunology*; D012756:Sheep / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology
chemical_list: D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D008697:Methane