Current Microbiology

, Volume 61, Issue 2, pp 125–131 | Cite as

Antimicrobial Effect of Red Clover (Trifolium pratense) Phenolic Extract on the Ruminal Hyper Ammonia-Producing Bacterium, Clostridium sticklandii



Ruminal proteolysis and subsequent amino acid degradation represent considerable economic loss in ruminant production. The hyper ammonia-producing bacteria (HAB) are largely responsible for amino acid deamination in the rumen. HAB can be controlled with ionophores, but they are also susceptible to antimicrobial plant secondary metabolites. Red clover (Trifolium pratense) is rich in soluble phenolics, and it is also more resistant to proteolysis than other legumes. The goal of this study was to identify phenolic compounds from Trifolium pratense cultivar Kenland, and determine if any of the compounds possessed antimicrobial activity against the bovine HAB, Clostridium sticklandii SR. HPLC analysis revealed that clover tissues were rich in the isoflavonoids formononetin and biochanin A, particularly in plants left to wilt for 24 h. Biochanin A inhibited C. sticklandii in bioassays that employed thin-layer chromatography (TLC). Both clover extracts and biochanin A inhibited the growth of C. sticklandii in broth culture, but formononetin had no effect. These results indicate that clover phenolic compounds may have a role in preventing amino acid fermentation.



We thank Dr. Norman L. Taylor (University of Kentucky, Department of Plant and Soil Science) for providing access to the clover plots used in this study, and for helpful discussions. This study was funded by the United States Department of Agriculture, Agricultural Research Service.


“Proprietary or brand names are necessary to report factually on available data; however, the USDA neither guarantees nor warrants the standard of the product, and the use of the name by the USDA implies no approval of the product, nor exclusion of others that may be suitable.”


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Copyright information

© US Government 2010

Authors and Affiliations

  1. 1.Forage-Animal Production Research UnitUSDA, Agricultural Research ServiceLexingtonUSA
  2. 2.Department of Animal and Food SciencesUniversity of KentuckyLexingtonUSA

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