Food, fibre and pharmaceuticals from animals
RESEARCH ARTICLE

# Mathematical formulae for accurate estimation of in vitro CH4 production from vented bottles

M. C. Hannah A C , P. J. Moate A , P. S. Alvarez Hess B , V. M. Russo A B , J. L. Jacobs A and R. J. Eckard B
+ Author Affiliations
- Author Affiliations

A Department of Economic Development, Jobs, Transport and Resources, Ellinbank Centre, 1301 Hazeldean Road, Ellinbank, Vic. 3821, Australia.

B Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Vic. 3010, Australia.

C Corresponding author. Email: murray.hannah@ecodev.vic.gov.au

Animal Production Science 56(3) 244-251 https://doi.org/10.1071/AN15577
Submitted: 14 September 2015  Accepted: 15 November 2015   Published: 9 February 2016

## Abstract

A widely used method for the in vitro measurement of fermentation parameters, total gas and methane (CH4) production (mL/g) from feed samples employs AnkomRF Technology with vented bottles, initially flushed with carbon dioxide (CO2). The volumes of gas accumulated between ventings, and the volumes vented, are calculated from the headspace pressure data. By using these data, it is possible to infer mathematically the quantity of CH4 produced, given just a single measurement of the CH4 mixing ratio measured in the headspace at the termination of fermentation. However, this calculation requires an additional assumption. Two possibilities for this are considered. We present mathematical formulae arising from each, and describe their sensitivity to violations of their respective assumptions. Also considered is a formula based on using N2 instead of CO2 as the flushing gas. It appears that, in practice, all three formulae are suitable for estimating CH4 production, with errors less than 3%.

Additional keywords: AnkomRF, methane, methodology.

## References

Cattani F, Tagliapietra F, Maccarana L, Hansen HH, Bailoni L, Schiavont S (2014) Technical note: In vitro total gas and methane production measurements from closed or vented rumen batch culture systems. Journal of Dairy Science 97, 1736–1741.
Technical note: In vitro total gas and methane production measurements from closed or vented rumen batch culture systems.CrossRef | 1:CAS:528:DC%2BC2cXktFKhtw%3D%3D&md5=4acab78a86b849c54f9bd7066e2254c7CAS |

Cone JW, Beuvink JMW, Rodrigues MAM (1994) Use and applications of an automated time related gas production test for the in vitro study of fermentation kinetics in the rumen. Revista Portuguesa de Zootecnia 1, 25–37.

Cornou C, Drejer Storm IML, Hindrichsen IK, Worgan H, Bakewell E, Yáñez Ruiz DR, Abecia L, Tagliapietra F, Cattani M, Ritz C, Hansen HH (2013) A ring test of a wireless in vitro gas production system. Animal Production Science 53, 585–592.
A ring test of a wireless in vitro gas production system.CrossRef |

Davies ZS, Mason D, Brooks AE, Griffith GW, Merry RJ, Theodorou MK (2000) An automated system for measuring gas production from forages inoculated with rumen fluid and its use in determining the effect of enzymes on grass silage. Animal Feed Science and Technology 83, 205–221.
An automated system for measuring gas production from forages inoculated with rumen fluid and its use in determining the effect of enzymes on grass silage.CrossRef | 1:CAS:528:DC%2BD3cXhslentbY%3D&md5=cd1f80fcb784c56750e3a95d9ca0e957CAS |

Dubois B, Tomkins NW, Kinley RD, Bai M, Seymour S, Paul NA, de Nys R (2013) Effect of tropical algae as additives on rumen in vitro gas production and fermentation characteristics. American Journal of Plant Sciences 4, 34–43.
Effect of tropical algae as additives on rumen in vitro gas production and fermentation characteristics.CrossRef | 1:CAS:528:DC%2BC2MXjsFart74%3D&md5=cb2a0cf330a77dc7d0039452a6a43b91CAS |

Getachew G, Robinson PH, DePeters EJ, Taylor SJ, Gisi DD, Higginbotham GE, Riordan TJ (2005) Methane production from commercial dairy rations estimated using an in vitro gas technique. Animal Feed Science and Technology 123–124, 391–402.
Methane production from commercial dairy rations estimated using an in vitro gas technique.CrossRef |

Machado L, Magnusson M, Paul NA, de Nys R, Tomkins N (2014) Effects of marine and freshwater macroalgae on in vitro total gas and methane production. PLoS One 9, e85289
Effects of marine and freshwater macroalgae on in vitro total gas and methane production.CrossRef | 24465524PubMed |

Machado L, Magnusson M, Paul NA, Kinley R, de Nys R, Tomkins N (2015) Dose-response effects of Asparagopsis taxiformis and Oedogonium sp. on in vitro fermentation and methane production. Journal of Applied Phycology
Dose-response effects of Asparagopsis taxiformis and Oedogonium sp. on in vitro fermentation and methane production.CrossRef |

Moate PJ, Clarke T, Davis LH, Laby RH (1997) Rumen gases and bloat in grazing dairy cows. Journal of Agricultural Science 129, 459–469.
Rumen gases and bloat in grazing dairy cows.CrossRef | 1:CAS:528:DyaK1cXot1OmtA%3D%3D&md5=c2d12a9e4c1c0a68b7c285bba6c33071CAS |

Moate PJ, Williams SRO, Grainger G, Hannah MC, Ponnampalam EN, Eckard RJ (2011) Influence of cold-pressed canola, brewers grains and hominy meal as dietary supplements suitable for reducing enteric methane emissions from lactating dairy cows. Journal of Animal Feed Science and Technology 166–167, 254–264.
Influence of cold-pressed canola, brewers grains and hominy meal as dietary supplements suitable for reducing enteric methane emissions from lactating dairy cows.CrossRef |

Moate PJ, Williams SRO, Torok VA, Hannah MC, Ribaux BE, Tavendale MH, Eckard RJ, Jacobs JL, Auldist MJ, Wales WJ (2014) Grape marc reduces methane emissions when fed to dairy cows. Journal of Dairy Science 97, 5073–5087.
Grape marc reduces methane emissions when fed to dairy cows.CrossRef | 1:CAS:528:DC%2BC2cXhtVCjur3E&md5=078818284e34aa3a309bb4ad6466718dCAS | 24952778PubMed |

Pell AN, Schofield P (1993) Computerized monitoring of gas production to measure forage digestion in vitro. Journal of Dairy Science 76, 1063–1073.
Computerized monitoring of gas production to measure forage digestion in vitro.CrossRef | 1:STN:280:DyaK3s3ltlWntQ%3D%3D&md5=87cc055940709c55768be9c1a120ae2eCAS | 8486838PubMed |

Pellikaan WF, Strigano E, Leenaars J, Bongers DJGM, van Laar-van Schuppen S, Plant J, Mueller-Harvey I (2011) Evaluating effects of tannins on extant and rate of in vitro gas and CH4 production using an automated pressure evaluation system (APES). Animal Feed Science and Technology 166–167, 377–390.
Evaluating effects of tannins on extant and rate of in vitro gas and CH4 production using an automated pressure evaluation system (APES).CrossRef |

Rymer C, Huntington JA, Williams BA, Givens DI (2005) In vitro cumulative gas production techniques: history, methodological considerations and challenges. Animal Feed Science and Technology 123–124, 9–30.
In vitro cumulative gas production techniques: history, methodological considerations and challenges.CrossRef |

Teshome K (2013) Effects of high starch–high fat and roughage diets of lambs on methane production using in-vitro gas production techniques. International Journal of Scientific & Engineering Research 4, 1553–1558.

Theodorou MK, Williams BA, Dhanoa MS, McAllan AB, France J (1994) A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds. Animal Feed Science and Technology 48, 185–197.
A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds.CrossRef |

Theodorou MK, Lowman RS, Davies ZS, Cuddeford D, Owen E (1998) Principles of techniques that rely on gas measurements in ruminant nutrition. In ‘In vitro techniques for measuring nutrient supply to ruminants’. BSAS occasional publication no. 22. (Eds ER Deaville, E Owen, AT Adesogen, C Rymer, JA Huntington, TLJ Lawrence) pp. 55–64. (BSAS: Edinburgh, UK)

Wilkins J (1974) Pressure transducer method for measuring gas production by microorganisms. Applied Microbiology 27, 135–140.

Xu M, Rinker M, McLeod KR, Harmon DL (2010) Yucca schidigera extract decreases in vitro methane production in a variety of forages and diets. Animal Feed Science and Technology 159, 18–26.
Yucca schidigera extract decreases in vitro methane production in a variety of forages and diets.CrossRef | 1:CAS:528:DC%2BC3cXosVGntr0%3D&md5=6c0e8991e91680c76edc77b936f51ac7CAS |

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