Addition of Purified Tannin Sources and Polyethylene Glycol Treatment on Methane Emission and Rumen Fermentation in Vitro

  • Anuraga Jayanegara Faculty of Animal Science, Bogor Agricultural University, Indonesia
  • H.P.S. Makkar Animal Production and Health Division, Food and Agriculture Organization of the United Nations, Rome, Italy
  • K. Becker Institute for Animal Production in the Tropics and Subtropics (480b), University of Hohenheim


The objectives of this experiment were (1) to observe the effects of purified tannins and polyethy-lene glycol (PEG) on in vitro rumen fermentation and methanogenesis, and (2) to assess the accuracy of volatile fatty acid (VFA) profiles in predicting methane emission. Hydrolysable and condensed tannins were extracted and purified from chestnut, sumach, mimosa and quebracho. Hay and concentrate mixture (70:30 w/w, 380 mg) was incubated in Hohenheim glass syringe containing 10 mL rumen liquor + 20 mL buffer. The purified tannins were injected into the syringes at a concentration of 1.0 mg/mL each, either without or with PEG 6,000 addition in three replicates. Results revealed that a decrease of methane emission (20%-27%) was observed when the purified tannins were added into basal diet as compared to control (P<0.05), and PEG addition increased methane emission (P<0.05). All purified tannins decreased total gas and total VFA production (P<0.05). The H2 recovery of the treatments ranged from 86.7% to 95.3%. Estimation of methane emission by using VFA profiles revealed an accurate result with a very low root mean square prediction error (1.75%). It is concluded that tannins mitigate methane emission while PEG neutralize such effect, and VFA profiles are accurate predictors of the emission.

Key words: tannin, polyethylene glycol, methane, rumen, stoichiometry


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Animut, G., R. Puchala, A. L. Goetsch, A. K. Patra, T. Sahlu, V. H. Varel, & J. Wells. 2008. Methane emission by goats consuming diets with different levels of condensed tannins from lespedeza. Anim. Feed Sci. Technol. 144:212-227.

Benchaar, C., & H. Greathead. 2011. Essential oils and opportunities to mitigate enteric methane emissions from ruminants. Anim. Feed Sci. Technnol. 166-167:338-355.

Bhatta, R., Y. Uyeno, K. Tajima, A. Takenaka, Y. Yabumoto, I. Nonaka, O. Enishi, & M. Kurihara. 2009. Difference in the nature of tannins on in vitro ruminal methane and volatile fatty acid production and on methanogenic archaea and protozoal populations. J. Dairy Sci. 92:5512-5522.

Castro Montoya, J., A. M. Bhagwat, N. Peiren, S. De Campeneere, D. De Baets & V. Fievez. 2011. Relationships between odd- and branched-chain fatty acid profiles in milk and calculated enteric methane proportion for lactating dairy cattle. Anim. Feed Sci. Technol. 166-167:596-602.

Cottle, D. J., J. V. Nolan, & S. G. Wiedemann. 2011. Ruminant enteric methane mitigation: a review. Anim. Prod. Sci. 51:491-514.

Hristov, A. N., J. Oh, J. L. Firkins, J. Dijkstra, E. Kebreab, G. Waghorn, H. P. S. Makkar, A. T. Adesogan, W. Yang, C. Lee, & P. J. Gerber. 2013. Mitigation of methane and nitrous oxide emissions from animal operations: I. A review of enteric methane mitigation options. J. Anim. Sci. 91:5045-5069.

Jayanegara, A., A. S. Tjakradidjaja, & T. Sutardi. 2006. Fermentabilitas dan kecernaan in vitro ransum limbah agroindustri yang disuplementasi kromium anorganik dan organik. Med. Pet. 29:54-62.

Jayanegara, A., & A. Sofyan. 2008. Penentuan aktivitas biologis tanin secara in vitro menggunakan Hohenheim gas test dengan polietilen glikol sebagai determinan. Med. Pet. 31:44-52.

Jayanegara, A., N. Togtokhbayar, H. P. S. Makkar, & K. Becker. 2009a. Tannins determined by various methods as predictors of methane production reduction potential of plants by an in vitro rumen fermentation system. Anim. Feed Sci. Technol. 150:230-237.

Jayanegara, A., H. P. S. Makkar, & K. Becker. 2009b. Emisi metana dan fermentasi rumen in vitro ransum hay yang mengandung tanin murni pada konsentrasi rendah. Med. Pet. 32:184-194.

Jayanegara, A., G. Goel, H. P. S. Makkar, & K. Becker. 2010. Reduction in methane emissions from ruminants by plant secondary metabolites: effects of polyphenols and saponins. In: Odongo, N. E., M. Garcia, & G. J. Viljoen (Eds). Sustainable Improvement of Animal Production and Health. Food and Agriculture Organization of the United Nations (FAO), Rome, Italy, pp. 151-157.

Jayanegara, A., F. Leiber, & M. Kreuzer. 2012. Meta-analysis of the relationship between dietary tannin level and methane formation in ruminants from in vivo and in vitro experiments. J. Anim. Physiol. Anim. Nutr. 96:365-375.

Jayanegara, A., Ikhsan, & T. Toharmat. 2013. Assessment of methane estimation from volatile fatty acid stoichiometry in the rumen in vitro. J. Indonesian Trop. Anim. Agric. 38:103-108.

Jayanegara, A., E. Wina, & J. Takahashi. 2014. Meta-analysis on methane mitigating properties of saponin-rich sources in the rumen in vitro: influence of addition levels and plant sources. Asian Australas. J. Anim. Sci. 27:1426-1435.

Kondo, M., Y. Hirano, N. Ikai, K. Kita, A. Jayanegara, & H. Yokota. 2014. Assessment of anti-nutritive activity of tannins in tea by-products based on in vitro ruminal fermentation. Asian Australas. J. Anim. Sci. 27:1571-1576.

Makkar, H. P. S., & K. Becker. 1994. Isolation of tannins from leaves of some trees and shrubs and their properties. J. Agric. Food Chem. 42:731-734.

Makkar, H. P. S., M. Blümmel, & K. Becker. 1995. Formation of complexes between polyvinyl pyrrolidones or polyethylene glycols and tannins, and their implication in gas production and true digestibility in in vitro techniques. Brit. J. Nutr. 73:897-913.

Makkar, H. P. S. 2003. Quantification of Tannins in Tree and Shrub Foliage, A Laboratory Manual. Kluwer Academic Publishers, Dordrecht, The Netherlands.

Monteny, G. J., A. Bannink, & D. Chadwick. 2006. Greenhouse gas abatement strategies for animal husbandry. Agric. Ecosyst. Environ. 112:163-170.

Morgavi, D. P., E. Forano, C. Martin, & C. J. Newbold. 2010. Microbial ecosystem and methanogenesis in ruminants. Animal 4:1024-1036.

Moss, A. R., J. P. Jouany, & J. Newbold. 2000. Methane production by ruminants: its contribution to global warming. Ann. Zootech. 49:231-253.

Mueller-Harvey, I. 2006. Unravelling the conundrum of tannins in animal nutrition and health. J. Sci. Food Agric. 86:2010-2037.

Tavendale, M. H., L. P. Meagher, D. Pacheco, N. Walker, G. T. Attwood, & S. Sivakumaran. 2005. Methane production from in vitro rumen incubations with Lotus pedunculatus and Medicago sativa, and effects of extractable condensed tannin fractions on methanogenesis. Anim. Feed Sci. Technol. 123-124:403-419.

Yogianto, A. Sudarman, E. Wina, & A. Jayanegara. 2014. Supplementation effects of tannin and saponin extracts to diets with different forage to concentrate ratio on in vitro rumen fermentation and methanogenesis. J. Indonesian Trop. Anim. Agric. (in press).

Yuliana, P., E. B. Laconi, E. Wina, & A. Jayanegara. 2014. Extraction of tannins and saponins from plant sources and their effects on in vitro methanogenesis and rumen fermentation. J. Indonesian Trop. Anim. Agric. (in press).