Technical Note| Volume 34, ISSUE 4, P390-396, August 2018

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Technical Note: The use of total gas collection for measuring methane production in vented in vitro systems


      In previous studies using Ankom GP vented in vitro systems (Ankom Technology, Macedon, NY), methane production was estimated from gas production (GP) and a single gas sample from the incubation vessel’s headspace at the end of the incubation. An accurate method requires measurement of methane in the incubation vessel’s headspace and in the gases vented over the incubation period. This research aimed to determine whether the act of collecting the gas vented from the Ankom GP system influenced the estimation by the Ankom GP system of the volume of gas produced or the composition of the gas produced. A method involving collecting gas through a long (304 cm), narrow (1 mm i.d.) gas line estimated greater GP than a control method not involving collection of gas. A method involving collecting gas through a short (22 cm), wide (4 mm i.d.) gas line did not affect headspace methane percentage or estimates of GP. It is concluded that in vitro methane production can be accurately estimated by using the Ankom GP system together with collection of vented gases into gas collection bags, but only if the Ankom GP system is connected to the gas collection bag via a short, wide gas line.

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        • Ankom
        Gas production service procedure validation test.
        (Accessed Dec. 10, 2017)
        • Bello N.M.
        • Kramer M.
        • Tempelman R.J.
        • Stroup W.W.
        • St-Pierre N.R.
        • Craig B.A.
        • Young L.J.
        • Gbur E.E.
        Short communication: On recognizing the proper experimental unit in animal studies in the dairy sciences.
        J. Dairy Sci. 2016; 99: 8871-8879
        • Cattani M.
        • Tagliapietra F.
        • Maccarana L.
        • Hansen H.H.
        • Bailoni L.
        • Schiavon S.
        Technical note: In vitro total gas and methane production measurements from closed or vented rumen batch culture systems.
        J. Dairy Sci. 2014; 97: 1736-1741
        • Dairy One
        Analytical Procedures.
        (Accessed Aug. 6, 2017)
        • Dubois B.
        • Tomkins N.W.
        • Kinley R.D.
        • Bai M.
        • Seymour S.
        • Paul N.A.
        • Nys R.
        Effect of tropical algae as additives on rumen in vitro gas production and fermentation characteristics.
        Am. J. Plant Sci. 2013; 4: 34-43
        • Getachew G.
        • Blümmel M.
        • Makkar H.
        • Becker K.
        In vitro gas measuring techniques for assessment of nutritional quality of feeds: A review.
        Anim. Feed Sci. Technol. 1998; 72: 261-281
        • Hannah M.C.
        • Moate P.J.
        • Alvarez-Hess P.A.
        • Russo V.M.
        • Jacobs J.L.
        • Eckard R.J.
        Mathematical formulae for accurate estimation of in vitro CH4 production from vented bottles.
        Anim. Prod. Sci. 2016; 56: 244-251
        • Laporte-Uribe J.A.
        The role of dissolved carbon dioxide in both the decline in rumen pH and nutritional diseases in ruminants.
        Anim. Feed Sci. Technol. 2016; 219: 268-279
        • Machado L.
        • Magnusson M.
        • Paul N.A.
        • de Nys R.
        • Tomkins N.
        Effects of marine and freshwater macroalgae on in vitro total gas and methane production.
        PLoS One. 2014; 9: e85289
        • Marten G.C.
        • Barnes R.F.
        Prediction of energy digestibility of forages with in vitro rumen fermentation and fungal enzyme systems.
        in: Pigden W. Balch C. Pages 61–128 in Standardization of Analytical Methodology for Feeds Workshop. Int. Dev. Res. Cent., Ottawa, Canada1980
        • Moate P.J.
        • Williams S.R.O.
        • Hannah M.C.
        • Eckard R.J.
        • Auldist M.J.
        • Ribaux B.E.
        • Jacobs J.L.
        • Wales W.J.
        Effects of feeding algal meal high in docosahexaenoic acid on feed intake, milk production, and methane emissions in dairy cows.
        J. Dairy Sci. 2013; 96: 3177-3188
        • NHMRC
        Australian Code of Practice for the Care and Use of Animals for Scientific Purposes.
        8th ed. 2013 (Accessed Dec. 12, 2017)
        • Pellikaan W.
        • Hendriks W.
        • Uwimana G.
        • Bongers L.
        • Becker P.
        • Cone J.
        A novel method to determine simultaneously methane production during in vitro gas production using fully automated equipment.
        Anim. Feed Sci. Technol. 2011; 168: 196-205
        • Ramin M.
        • Huhtanen P.
        Development of an in vitro method for determination of methane production kinetics using a fully automated in vitro gas system—A modelling approach.
        Anim. Feed Sci. Technol. 2012; 174: 190-200
        • Robinson P.
        • Mathews M.C.
        • Fadel J.
        Influence of storage time and temperature on in vitro digestion of neutral detergent fibre at 48h, and comparison to 48h in sacco neutral detergent fibre digestion.
        Anim. Feed Sci. Technol. 1999; 80: 257-266
        • Russo V.M.
        • Jacobs J.L.
        • Hannah M.C.
        • Moate P.J.
        • Dunshea F.R.
        • Leury B.J.
        In vitro evaluation of the methane mitigation potential of a range of grape marc products.
        Anim. Prod. Sci. 2017; 57: 1437-1444
        • Shiotani T.
        • Yamane T.
        A horizontal packed-bed bioreactor to reduce CO2 gas holdup in the continuous production of ethanol by immobilized yeast cells.
        Appl. Microbiol. Biotechnol. 1981; 13: 96-101
        • Vennard J.K.
        • Street R.L.
        Fluid flow in pipes.
        in: Vennard J.K. Page 359 in Elementary Fluid Mechanics. John Wiley Sons, New York, NY1982
        • Wang M.
        • Wang R.
        • Tang S.X.
        • Tan Z.L.
        • Zhou C.S.
        • Han X.F.
        • Kang J.H.
        Comparisons of manual and automated incubation systems: Effects of venting procedures on in vitro ruminal fermentation.
        Livest. Sci. 2016; 184: 41-45
        • Xu M.
        • Rinker M.
        • McLeod K.
        • Harmon D.
        Yucca schidigera extract decreases in vitro methane production in a variety of forages and diets.
        Anim. Feed Sci. Technol. 2010; 159: 18-26