Rumen Microbial Population in the In vitro Fermentation of Different Ratios of Forage and Concentrate in the Presence of Whole Lerak (Sapindus rarak) Fruit Extract

Suharti, Sri;Astuti, Dewi Apri;Wina, Elizabeth;Toharmat, Toto

  • Received : 2010.11.10
  • Accepted : 2011.01.16
  • Published : 2011.08.01


This experiment was designed to investigate the effect of lerak extract on the dynamic of rumen microbes in the in vitro fermentation of diet with different ratios of forage and concentrate. In vitro fermentation was conducted according to the method of Tilley and Terry (1963). The design of experiment was a factorial block design with 2 factors. The first factor was the ratio of forage and concentrate (90:10, 80:20, and 70:30 w/w) and the second factor was the level of lerak extract (0, 0.6, and 0.8 mg/ml). Total volatile fatty acid (VFA) concentration, proportional VFA and NH3 concentration were measured at 4 h incubation. Protozoal numbers in the buffered rumen fluid after 4 and 24 h of incubation were counted under a microscope. Bacterial DNAs of buffered rumen fluid were isolated from incubated samples after 24 h of incubation using a QiaAmp kit. Total bacteria, Fibrobacter succinogenes, Ruminococcus albus, and Prevotella ruminicola were quantified using real time polymerase chain reaction (PCR). Lerak extract markedly reduced protozoal numbers in buffered rumen fluid of all diets after 24 h of incubation. Total bacteria did not change with lerak extract addition. While no difference in F. succinogenes was found, there was a slight increase in R. albus number and a significant enhancement in P. ruminicola number by increasing the level of lerak extract in all diets. Propionate concentration significantly increased in the presence of lerak extract at level 0.8 mg/ml. It was concluded that the addition of lerak extract could modify rumen fermentation and had positive effects on rumen microbes.


Sapindus rarak;Rumen Microbial;Fermentation;Forage to Concentrate Ratio


  1. Benchaar, C., T. A. McAllister and P. Y. Choulnard. 2008. Digestion, rumen fermentation, ciliate protozoal populations, and milk production from dairy cows fed cinnamaldehyde, quebracho condensed tannin, or Yucca schidigera saponin extracts. J. Dairy Sci. 91:4786-4777.
  2. Denman, S. E. and C. S. McSweeney. 2006. Developmentofa realtimePCR assay formonitoring anaerobic fungal and cellulolytic bacterial populationswithin the rumen. FEMS Microbiol. Ecol. 58:572-582.
  3. Francis, G., Z. Keem, H. P. S. Makkar and K. Becker. 2002. The biological action of saponins in animal systems: a review. Br. J. Nutr. 88:587-605.
  4. Guo, Y., J. Liu, Y. Lu, W. Y. Zhu, S. E. Denman and C. S. McSweeney. 2008. Effect of tea saponin on methanogenesis, microbial community structure and expression of mcrA gene, in cultures of rumen micro-organisms. Lett. Appl. Microbiol. 47:421-426.
  5. Goel, G., H. P. S. Makkar and K. Becker. 2008. Changes in microbial community structure, methanogenesis and rumen fermentation in response to saponin-rich fractions from different plant materials. J. Appl. Microbiol. 105:770-777.
  6. Gutierrez, J. 2007. Observations on bacterial feeding by the rumen ciliate Isotricha prostoma. J. Eukaryotic Microbiol. 5:122-126.
  7. Hamburger, M. I. Slacanin, K. Hostettmann, W. Dyatmiko and Sutarjadi. 1992. Acetylated saponins in molluscicidal activity from Sapindus rarak: Unambiguous structure determination by proton nuclear magnetic resonance and quantitative analysis. Phytochem. Anal. 3:231-237.
  8. Hess, H. D., M. Kreuzer, T. E. Diaz, C. E. Lascano, J. E. Carulla and C. R. Solvia. 2003. Saponin rich tropical fruits affect fermentation and methanogenesis in faunated and defaunated fluid. Anim. Feed Sci. Technol. 109:79-94.
  9. Hsu, J. T., G. C. Fahey, N. R. Merchen and R. I. Mackie. 1991. Effects of defaunation and various nitrogen supplementation regimens on microbial numbers and activity in the rumen of sheep. J. Anim. Sci. 69:1279-1289.
  10. Hu W., J. Liu, J. Ye, Y. Wu and Y. Guo. 2005. Effect of tea saponin on rumen fermentation in vitro. Anim. Feed Sci. Technol. 120:333-339.
  11. Ivan, M., L. Neill and T. Entz. 2000. Rumen fermentation and duodenal flow following progressive inoculations of fauna-free wethers with major individual species of ciliate protozoa or total fauna. J. Anim. Sci. 78:750-759.
  12. Kamra, D. N. 2005. Rumen microbial ecosystem. Curr. Sci. 89:1- 12.
  13. Karnati, S. K. R., Z. Yu and J. L. Firkins. 2009. Investigating unsaturated fat, monensin, or bromoethanesulfonate in continuous cultures retaining rumen protozoa. II. Interaction of treatment and presence of protozoa on prokaryotic communities. J. Dairy Sci. 92:3861-3873.
  14. 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. Lett. 204:361- 366.
  15. Makkar, H. P. S. and K. Becker. 1997. Degradation of quillaja saponins by mixed culture of rumen microbes. Lett. Appl. Microbiol. 25:243-245.
  16. Mao, H., J. Wang, Y. Zhou and J. Liu. 2010. Effects of addition of tea saponins and soybean oil on methane production, fermentation and microbial population in the rumen of growing lambs. Anim. Feed Sci. Technol. 129:56-62.
  17. Muetzel, S., R. Akpagloh and K. Becker. 2005. Sapindus rarak saponins do not affect rumen protein degradation in vitro. Proc. Soc. Nutr. Physiol. 14.
  18. Murray, R. K., D. K. Granner and V. W. Rodwell. 2006. Harper's Illustrated Biochemistry, 27TH Edition. The McGraw-Hill Companies, USA.
  19. Tajima, K., R. I. Aminov, T. Nagamine, H. Matsui, M. Nakamura and Y. Benno. 2001. Diet-dependent shifts in the bacterial population of the rumen revealed with real-time PC. Appl. Environ. Microbiol. 67:2766-2774.
  20. Teferedegne, B. 2000. New perspectives on the use of tropical plants to improve ruminant nutrition. Proc. Nutr. Soc. 59:209- 214.
  21. Thalib, A., Y. Widiawati, H. Hamid, D. Suherman and M. Sabrani. 1995. The effects of saponins from Sapindusrarak fruit on rumen microbes and host animal growth. Ann Zootech. 44 (Suppl.):161.
  22. Tilley, J. M. A. and R. A. Terry. 1963. A two stage technique for the in vitro digestion of forage. J. Br. Grassland Soc. 18:104- 111.
  23. Wallace, R. J., L. Arthaud and C. J. Newbold. 1994. Influence of Yucca shidigera extract on rumen ammonia concentrations and rumen microorganisms. Appl. Environ. Microbiol. 60:1762- 1767.
  24. Wallace, R. J. and C. A. McPherson. 1987. Factors affecting the rate of breakdown of bacterial protein in rumen fluid. Br. J. Nutr. 5:313-323.
  25. Wang, Y., T. A. McAllister, C. J. Newbold, L. M. Rodea, P. R. Cheeke and K. J. Cheng. 1998. Effects of Yucca schidigera extract on fermentation and degradation of steroidal saponins in the rumen simulation technique (RUSITEC). Anim. Feed Sci. Technol. 74:143-153.
  26. Wang, Y., T. A. McAllister, L. J. Yanke, Z. J. Xu, P. R. Cheeke and K. J. Cheng. 2000. In vitro effects of steroidal saponins from Yucca schidigera extract on rumen microbial protein synthesis and rumen fermentation. J. Sci. Food Agric. 80:2214-2122.
  27. Wina, E., S. Muetzel, E. M. Hoffmann, H. P. S. Makkar and K. Becker. 2005. Saponins containing methanol extract of sapindus rarak affect microbial fermentation, microbial activity and microbial community structure in vitro. Anim. Feed Sci. Technol. 121:159-174.
  28. Wina, E., S. Muetzel and K. Becker. 2006. Effects of daily and interval feeding of Sapindus rarak saponins on protozoa, rumen fermentation parameters and digestibility in sheep. Asian-Aust. J. Anim. Sci. 19:1580-1587.
  29. Yost, W. M., J. W. Young, S. P. Schmidt and A. D. Mcgilliard. 1977. Gluconeogenesis in ruminants: propionic acid production from a high-grain diet fed to cattle. J. Nutr. 107: 2036-2043.

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