Effects of Flavonoid-rich Plant Extracts on In vitro Ruminal Methanogenesis, Microbial Populations and Fermentation Characteristics

  • Kim, Eun T. (National Institute of Animal Science, RDA) ;
  • Guan, Le Luo (Department of Agricultural, Food and Nutritional Science, University of Alberta) ;
  • Lee, Shin J. (Division of Applied Life Science (BK21+, IALS), Gyeongsang National University) ;
  • Lee, Sang M. (National Institute of Animal Science, RDA) ;
  • Lee, Sang S. (Department of Animal Science and Technology, Sunchon National Uuniversity) ;
  • Lee, Il D. (Division of Applied Life Science (BK21+, IALS), Gyeongsang National University) ;
  • Lee, Su K. (Division of Applied Life Science (BK21+, IALS), Gyeongsang National University) ;
  • Lee, Sung S. (Division of Applied Life Science (BK21+, IALS), Gyeongsang National University)
  • Received : 2014.09.06
  • Accepted : 2014.11.04
  • Published : 2015.04.01


The objective of this study was to evaluate the in vitro effects of flavonoid-rich plant extracts (PE) on ruminal fermentation characteristics and methane emission by studying their effectiveness for methanogenesis in the rumen. A fistulated Holstein cow was used as a donor of rumen fluid. The PE (Punica granatum, Betula schmidtii, Ginkgo biloba, Camellia japonica, and Cudrania tricuspidata) known to have high concentrations of flavonoid were added to an in vitro fermentation incubated with rumen fluid. Total gas production and microbial growth with all PE was higher than that of the control at 24 h incubation, while the methane emission was significantly lower (p<0.05) than that of the control. The decrease in methane accumulation relative to the control was 47.6%, 39.6%, 46.7%, 47.9%, and 48.8% for Punica, Betula, Ginkgo, Camellia, and Cudrania treatments, respectively. Ciliate populations were reduced by more than 60% in flavonoid-rich PE treatments. The Fibrobacter succinogenes diversity in all added flavonoid-rich PE was shown to increase, while the Ruminoccocus albus and R. flavefaciens populations in all PE decreased as compared with the control. In particular, the F. succinogenes community with the addition of Birch extract increased to a greater extent than that of others. In conclusion, the results of this study showed that flavonoid-rich PE decreased ruminal methane emission without adversely affecting ruminal fermentation characteristics in vitro in 24 h incubation time, suggesting that the flavonoid-rich PE have potential possibility as bio-active regulator for ruminants.


Supported by : Rural Development Administration


  1. Baker, S. K. 1999. Rumen methanogens, and inhibition of methanogenesis. Aust. J. Agric. Res. 50:1293-1298.
  2. Balcells, J., A. Aris, A. Serrano, A. R. Seradj, J. Crespo, and M. Devant. 2012. Effects of an extract of plant flavonoids (Bioflavex) on rumen fermentation and performance in heifers fed high-concentrate diets. J. Anim. Sci. 90:4975-4984.
  3. Becker, P. M., P. G. Van Wikselaar, M. C. R. Franssen, R. C. H. De Vos, R. D. Hall, and J. Beekwilder. 2014. Evidence for a hydrogen-sink mechanism of (+) catechin-mediated emission reduction of the ruminant greenhouse gas methane. Metabolomics 10:179-189.
  4. Benchaar, C., H. V. Petit, R. Berthiaume, D. R. Ouellet, J. Chiquette, and P. Y. Chouinard. 2007. Effects of essential oils on digestion, ruminal fermentation, rumen microbial populations, milk production, and milk composition in dairy cows fed alfalfa silage or corn silage. J. Dairy Sci. 90:886-897.
  5. Bhatta, R., Y. Uyeno, K. Tajima, A. Takenaka, Y. Yabumoto, I. Nonaka, O. Enishi, and 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.
  6. Bodas, R., S. Lopez, M. Fernandez, R. Garcia-Gonzalez, A. B. Rodriguez, R. J. Wallace, and J. S. Gonzalez. 2008. In vitro screening of the potential of numerous plant species as antimethanogenic feed additives for ruminants. Anim. Feed Sci. Technol. 145:245-258.
  7. Cushnie, T. P. T. and A. J. Lamb. 2005. Antimicrobial activity of flavonoids. Int. J. Antimicrob. Agents 26:343-356.
  8. Denman, S. E. and C. S. McSweeney. 2005. Quantitative (realtime) PCR. In Methods in Gut Microbial Ecology for Ruminants (Eds. H. P. S. Makkar and C. S. McSweeney). Springer, Dordrecht, The Netherlands. pp. 105-115.
  9. Denman, S. E. and C. S. McSweeney. 2006. Development of a Real-Time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen. FEMS. Microbiol. Ecol. 58:572-582.
  10. Denman, S. E., N. W. Tomkins, and C. S. McSweeney. 2007. Quantitation and diversity analysis of ruminal methanogenic populations in response to the antimethanogenic compound bromochloromethane. FEMS Microbiol. Ecol. 62:313-322.
  11. Dohme, F., A. Machmuller, A. Wasserfallen, and M. Kreuzer. 2001. Ruminal methanogenesis as influenced by individual fatty acids supplemented to complete ruminant diets. Lett. Appl. Microbiol. 32:47-51.
  12. Ellis, J. L., E. Kebreab, N. E. Odongo, B. W. McBride, E. K. Okine, and J. France. 2007. Prediction of methane production from dairy and beef cattle. J. Dairy Sci. 90:3456-3467.
  13. Jia, Z., M. Tang, and J. Wu. 1999. The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals. Food Chem. 64:555-559.
  14. Johnson, K. A. and D. E. Johnson. 1995. Methane emissions from cattle. J. Anim. Sci. 73:2483-2492.
  15. Koike, S. and Y. Kobayashi. 2001. Development and use of competitive PCR assays for the rumen cellulolytic bacteria: Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens. FEMS Microbiol. Ecol. 204:361-366.
  16. Latham, M. J. and M. J. Wolin. 1977. Fermentation of cellulose by Ruminococcus flavefaciens in the presence and absence of Methanobacterium ruminantium. Appl. Environ. Microbiol. 34:297-301.
  17. McDougall, E. I. 1948. Studies on ruminant saliva. 1. The composition and output of sheep's saliva. Biochem. J. 43:99-109.
  18. Ntaikou, I., H. N. Gavala, M. Kornaros, and G. Lyberatos. 2008. Hydrogen production from sugars and sweet sorghum biomass using Ruminococcus albus. Int. J. Hydrogen. Energy 33:1153-1163.
  19. Oskoueian, E., N. Abdullah, and A. Oskoueian. 2013. Effects of flavonoids on rumen fermentation activity, methane production, and microbial population. Biomed Res. Int. Article ID 349129, 8 pages. Doi:10.1155/2013/349129.
  20. Patra, A. K., D. N. Kamra, and N. Agarwa. 2006. Effect of plant extracts on in vitro methanogenesis, enzyme activities and fermentation of feed in rumen liquor of buffalo. Anim. Feed Sci. Technol. 128:276-291.
  21. Patra, A. K. and J. Saxena. 2010. A new perspective on the use of plant secondary metabolites to inhibit methanogenesis in the rumen. Phytochemistry 71:1198-1222.
  22. Pen, B., C. Sar, B. Mwenya, K. Kuwaki, R. Morikawa, and J. Takahashi. 2006. Effects of Yucca schidigera and Quillaja saponaria extracts on in vitro ruminal fermentation and methane emission. Anim. Feed Sci. Technol. 129:175-186.
  23. SAS Institute. 2002. SAS User's Guide. SAS Institute Inc., Cary, NC, USA.
  24. Skillman, L. C., P. N. Evans, C. Strompl, and K. N. Joblin. 2006. 16S rDNA directed PCR primers and detection of methanogens in the bovine rumen. Lett. Appl. Microbiol. 42:222-228.
  25. Skillman, L. C., A. F. Toovey, A. J. Williams, and A. G. Wright. 2006. Development and validation of a real-time PCR method to quantify rumen protozoa and examination of variability between Entodinium populations in sheep offered a hay-based diet. Appl. Environ. Microbiol. 72:200-206.
  26. Tedesco, D., A. Tava, S. Galletti, M. Tameni, G. Varisco, A. Costa, and S. Steidler. 2004. Effects of silymarin, a natural hepatoprotector, in periparturient dairy cows. J. Dairy Sci. 87:2239-2247.
  27. Theodorou, M. K., B. A. Williams, M. S. Dhanoa, A. B. McAllan, and J. France. 1994. A simple gas production method using a pressure transducer to determine the fermentation kinetics of ruminant feeds. Anim. Feed Sci. Technol. 48:185-197.
  28. Velioglu, Y. S., G. Mazza, L. Cao, and B. D. Oomah. 1998. Antioxidant activity and total phenolics in selected fruit, vegetables, and grain products. J. Agric. Food Chem. 46:4113-4117.
  29. Zhou, Z., Q. Meng, and Z. Yu. 2011. Effects of methanogenic inhibitors on methane production and abundances of methanogens and cellulolytic bacteria in in vitro ruminal cultures. Appl. Environ. Microbiol. 77:2634-2639.

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