Animal Production Science Animal Production Science Society
Food, fibre and pharmaceuticals from animals
RESEARCH ARTICLE

Dose-response effect of nitrate on hydrogen distribution between rumen fermentation end products: an in vitro approach

J. Guyader A C , M. Tavendale B , C. Martin A and S. Muetzel B
+ Author Affiliations
- Author Affiliations

A INRA, UMR1213 Herbivores, F-63122 Saint-Genès-Champanelle, France; and Clermont Université, VetAgro Sup, UMR1213 Herbivores, BP 10448, F-63000, Clermont-Ferrand, France.

B AgResearch Grasslands, Private Bag 11-008, Palmerston North 4442, New Zealand.

C Corresponding author. Email: jess.guyader@hotmail.fr

Animal Production Science 56(3) 224-230 https://doi.org/10.1071/AN15526
Submitted: 1 September 2015  Accepted: 2 November 2015   Published: 9 February 2016

Abstract

The objective of this work was to study the in vitro dose-response effect of nitrate (0, 1, 2, 4 and 6 mM) on metabolic hydrogen distribution between rumen fermentation end products. Three 48-h incubations were conducted using bovine rumen contents as an inoculum, and a mixture of hay and concentrate (50 : 50) as a substrate. Total gas production and composition (methane and hydrogen) were automatically analysed throughout the incubations. Volatile fatty acid and ammonium concentrations were analysed from samples taken after 48 h of incubation. Total gas production was decreased with the highest dose of nitrate (P = 0.002). Methane emissions linearly decreased as the nitrate dose increased (P = 0.005). Kinetics of methane emissions showed that metabolic hydrogen removal via nitrate reduction occurred mainly during the first 10 h of incubation. Gaseous hydrogen production was similar among treatments, despite higher hydrogen emissions for nitrate concentrations >4 mM. Concentrations and proportions of volatile fatty acids were not affected by treatments. The proportion of unaccounted metabolic hydrogen was positive for all treatments, and tended to linearly increase as the nitrate dose increased. In this in vitro work, we confirmed that nitrate is an efficient methane-mitigating compound in the rumen. We also suggest that nitrate or its reduced forms have a direct inhibiting effect towards methanogens, as indicated by the release of gaseous hydrogen and the high efficiency of methane mitigation. However, high nitrate concentrations also decrease overall fermentation.

Additional keywords: CH4, gas production, hydrogen recovery, ruminant, VFA.


References

Attwood GT, Klieve AV, Ouwerkerk D, Patel BKC (1998) Ammonia-hyperproducing bacteria from New Zealand ruminants. Applied and Environmental Microbiology 64, 1796–1804.

Bozic AK, Anderson RC, Carstens GE, Ricke SC, Callaway TR, Yokoyama MT, Wang JK, Nisbet DJ (2009) Effects of the methane-inhibitors nitrate, nitroethane, lauric acid, Lauricidin and the Hawaiian marine algae Chaetoceros on ruminal fermentation in vitro. Bioresource Technology 100, 4017–4025.
Effects of the methane-inhibitors nitrate, nitroethane, lauric acid, Lauricidin and the Hawaiian marine algae Chaetoceros on ruminal fermentation in vitro.CrossRef | 1:CAS:528:DC%2BD1MXmtVelsbY%3D&md5=c64893448b75762d2176c84f97e89e13CAS | 19362827PubMed |

Chaney AL, Marbach EP (1962) Modified reagents for determination of urea and ammonia. Clinical Chemistry 8, 130–132.

Czerkawski JW (1986) ‘An introduction to rumen studies.’ (Pergamon Press: New York)

France J, Dijkstra J, Dhanoa MS, Lopez S, Bannink A (2000) Estimating the extent of degradation of ruminant feeds from a description of their gas production profiles observed in vitro: derivation of models and other mathematical considerations. British Journal of Nutrition 83, 143–150.
Estimating the extent of degradation of ruminant feeds from a description of their gas production profiles observed in vitro: derivation of models and other mathematical considerations.CrossRef | 1:CAS:528:DC%2BD3cXhsVymtbY%3D&md5=ea3b4276bff65a900515c6e00b16049cCAS | 10743493PubMed |

Guo WS, Schaefer DM, Guo XX, Ren LP, Meng QX (2009) Use of nitrate-nitrogen as a sole dietary nitrogen source to inhibit ruminal methanogenesis and to improve microbial nitrogen synthesis in vitro. Asian-Australasian Journal of Animal Sciences 22, 542–549.
Use of nitrate-nitrogen as a sole dietary nitrogen source to inhibit ruminal methanogenesis and to improve microbial nitrogen synthesis in vitro.CrossRef | 1:CAS:528:DC%2BD1MXlsVeitLw%3D&md5=8149ab644f49c3bc7017be10322a9eb1CAS |

Guyader J, Silberberg M, Popova M, Seradj AR, Morgavi DP, Martin C (2014) Dietary nitrates decrease methane emission by inhibiting rumen methanogenic archaea without influencing nitrate reducing bacteria. In ‘Proceedings of the 9th Joint Rowett/INRA symposium, gut microbiology: from sequence to function’. p. 13. (Rowett Institute of Nutrition and Health, University of Aberdeen: Aberdeen, UK)

Guyader J, Eugène M, Meunier B, Doreau M, Morgavi DP, Silberberg M, Rochette Y, Gérard C, Loncke C, Martin C (2015) Additive methane-mitigating effect between linseed oil and nitrate fed to cattle. Journal of Animal Science 93, 3564–3577.
Additive methane-mitigating effect between linseed oil and nitrate fed to cattle.CrossRef | 1:CAS:528:DC%2BC2MXht1Ols7fN&md5=04b1c9c93da0074846e43c590517e35aCAS | 26440025PubMed |

Hegarty RS, Gerdes R (1999) Hydrogen production and transfer in the rumen. Recent Advances in Animal Nutrition 12, 37–44.

Janssen PH (2010) Influence of hydrogen on rumen methane formation and fermentation balances through microbial growth kinetics and fermentation thermodynamics. Animal Feed Science and Technology 160, 1–22.
Influence of hydrogen on rumen methane formation and fermentation balances through microbial growth kinetics and fermentation thermodynamics.CrossRef | 1:CAS:528:DC%2BC3cXhtV2itLvF&md5=8b633b0832b1c0985b793ee6af3ece67CAS |

Lee C, Beauchemin KA (2014) A meta-analysis of effects of feeding nitrate on toxicity, production, and enteric methane emissions in ruminants. In ‘Proceedings of the joint annual meeting, linking animal science and animal agriculture: meeting the global demands of 2050’. pp. 845–846. (American Dairy Science Association and American Society of Animal Science: Kansas City, MO)

Mills JA, Dijkstra J, Bannink A, Cammell SB, Kebreab E, France J (2001) A mechanistic model of whole-tract digestion and methanogenesis in the lactating dairy cow: model development, evaluation, and application. Journal of Animal Science 79, 1584–1597.

Mould FL, Morgan R, Kliem KE, Krystallidou E (2005) A review and simplification of the in vitro incubation medium. Animal Feed Science and Technology 123–124, 155–172.
A review and simplification of the in vitro incubation medium.CrossRef |

Muetzel S, Hunt C, Tavendale MH (2014) A fully automated incubation system for the measurement of gas production and gas composition. Animal Feed Science and Technology 196, 1–11.
A fully automated incubation system for the measurement of gas production and gas composition.CrossRef | 1:CAS:528:DC%2BC2cXhtlamsb3I&md5=38be41c59512b6ff82ec66c2fec4e89eCAS |

Newbold JR, van Zijderveld SM, Hulshof RBA, Fokkink WB, Leng RA, Terencio P, Powers WJ, van Adrichem PSJ, Paton ND, Perdok HB (2014) The effect of incremental levels of dietary nitrate on methane emissions in Holstein steers and performance in Nelore bulls. Journal of Animal Science 92, 5032–5040.
The effect of incremental levels of dietary nitrate on methane emissions in Holstein steers and performance in Nelore bulls.CrossRef | 1:CAS:528:DC%2BC2MXisFWis7Y%3D&md5=9e964395af6d2ee6cbe1900e80e25202CAS | 25349351PubMed |

Patra AK, Yu Z (2013) Effective reduction of enteric methane production by a combination of nitrate and saponin without adverse effect on feed degradability, fermentation, or bacterial and archaeal communities of the rumen. Bioresource Technology 148, 352–360.
Effective reduction of enteric methane production by a combination of nitrate and saponin without adverse effect on feed degradability, fermentation, or bacterial and archaeal communities of the rumen.CrossRef | 1:CAS:528:DC%2BC3sXhs1Wisr7J&md5=9903999e8ebbf014db7aa5f1e5b29e43CAS | 24063817PubMed |

Patra AK, Yu Z (2014) Combinations of nitrate, saponin, and sulfate additively reduce methane production by rumen cultures in vitro while not adversely affecting feed digestion, fermentation or microbial communities. Bioresource Technology 155, 129–135.
Combinations of nitrate, saponin, and sulfate additively reduce methane production by rumen cultures in vitro while not adversely affecting feed digestion, fermentation or microbial communities.CrossRef | 1:CAS:528:DC%2BC2cXmslWqtbY%3D&md5=b1def954d9c4cc732e2e95fbdd6fd033CAS | 24440491PubMed |

Sakthivel PC, Kamra DN, Agarwal N, Chaudhary LC (2012) Effect of sodium nitrate and nitrate reducing bacteria on in vitro methane production and fermentation with buffalo rumen liquor. Asian-Australasian Journal of Animal Sciences 25, 812–817.
Effect of sodium nitrate and nitrate reducing bacteria on in vitro methane production and fermentation with buffalo rumen liquor.CrossRef | 1:CAS:528:DC%2BC38XhtFCjtrzN&md5=e23cabbb1fa2f9058d70c34577c90890CAS | 25049631PubMed |

Sar C, Santoso B, Mwenya B, Gamoa Y, Kobayashi T, Morikawa R, Kimura K, Mizukoshi H, Takahashi J (2004) Manipulation of rumen methanogenesis by the combination of nitrate with β1–4 galacto-oligosaccharides or nisin in sheep. Animal Feed Science and Technology 115, 129–142.
Manipulation of rumen methanogenesis by the combination of nitrate with β1–4 galacto-oligosaccharides or nisin in sheep.CrossRef | 1:CAS:528:DC%2BD2cXks1eqs7c%3D&md5=517f8c3d9ac809f492bd56aaab0ff615CAS |

Shi C, Meng Q, Hou X, Ren L, Zhou Z (2012) Response of ruminal fermentation, methane production and dry matter digestibility to microbial source and nitrate addition level in an in vitro incubation with rumen microbes obtained from wethers. Journal of Animal and Veterinary Advances 11, 3334–3341.
Response of ruminal fermentation, methane production and dry matter digestibility to microbial source and nitrate addition level in an in vitro incubation with rumen microbes obtained from wethers.CrossRef |

Ungerfeld EM (2015) Shifts in metabolic hydrogen sinks in the methanogenesis-inhibited ruminal fermentation: a meta-analysis. Frontiers in Microbiology 6, 1–17.

Ungerfeld EM, Rust SR, Burnett R (2006) Effects of butyrate precursors on electron relocation when methanogenesis is inhibited in ruminal mixed cultures. Letters in Applied Microbiology 42, 567–572.

van Zijderveld SM, Gerrits WJJ, Apajalahti JA, Newbold JR, Dijkstra J, Leng RA, Perdok HB (2010) Nitrate and sulfate: effective alternative hydrogen sinks for mitigation of ruminal methane production in sheep. Journal of Dairy Science 93, 5856–5866.
Nitrate and sulfate: effective alternative hydrogen sinks for mitigation of ruminal methane production in sheep.CrossRef | 1:CAS:528:DC%2BC3MXjs1Kis7Y%3D&md5=c84de18e60e1d808cc7a5d43ed5f5cd6CAS | 21094759PubMed |

van Zijderveld SM, Gerrits WJJ, Dijkstra J, Newbold JR, Hulshof RBA, Perdok HB (2011) Persistency of methane mitigation by dietary nitrate supplementation in dairy cows. Journal of Dairy Science 94, 4028–4038.
Persistency of methane mitigation by dietary nitrate supplementation in dairy cows.CrossRef | 1:CAS:528:DC%2BC3MXpsVylur4%3D&md5=4b50ef913e6ff901e51ebf368f5f53b0CAS | 21787938PubMed |

Veneman JB, Muetzel S, Hart KJ, Faulkner CL, Moorby JM, Molano G, Perdok HB, Newbold JR, Newbold CJ (2014) Dietary nitrate but not linseed oil decreases methane emissions in two studies with lactating dairy cows. In ‘Proceedings of the livestock, climate change and food security conference’. p. 38. (Madrid, Spain)

Wolin M, Miller T, Stewart C (1997) Microbe–microbe interactions. In ‘The rumen microbial ecosystem’. (Eds PN Hobson, CS Stewart) pp. 467–491. (Chapman & Hall: London)

Zhou Z, Meng Q, Yu Z (2011) Effects of methanogenic inhibitors on methane production and abundances of methanogens and cellulolytic bacteria in in vitro ruminal cultures. Applied and Environmental Microbiology 77, 2634–2639.
Effects of methanogenic inhibitors on methane production and abundances of methanogens and cellulolytic bacteria in in vitro ruminal cultures.CrossRef | 1:CAS:528:DC%2BC3MXhtVWktLzJ&md5=4d1768c87df18cfbbd34683d2c0a4257CAS | 21357427PubMed |



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