Biochanin A Inhibits Ruminal Nitrogen-Metabolizing Bacteria and Alleviates the Decomposition of Amino Acids and Urea In Vitro

SIMPLE SUMMARY: Plant bioactive compounds have been chosen as alternative antibiotic to promote animal productivity. Biochanin A is a type of naturally occurring bioactive compound. It is O-methylated isoflavone and is found in red clover, alfalfa sprouts, and other legumes. The aim of this study was to determine the effect of biochanin A on rumen microbial fermentation and composition. The results show that biochanin a increases microbial gas production, but has no effect on volatile fatty acid (VFA) production. Microbial urease activity was inhibited by Biochanin A with the IC50 of 320 nM. Biochanin A also inhibited the degradation rate of Val, Lys, Met, Leu and total amino acids, respectively. The inhibition of urease activity and amino acid decomposition by biochanin A resulted in a reduction in ammonia. The 16S rRNA gene sequencing showed that biochanin A reduced the abundance of proteolytic bacteria Prevotella and Streptococcus. Therefore, biochanin A reduced the production of ammonia by inhibiting proteolytic bacteria and its decomposition of urea and amino acids activity. ABSTRACT: Biochanin A is a naturally occurring flavonoid compound that is found in plant species such as red clover (Trifolium pretense) and alfalfa (Medicago sativa). Flavonoids have been reported to regulate ruminal fermentation, and the objective of this study was to evaluate the effects of biochanin A on ruminal microbial composition and nitrogen metabolism. The experiment was performed by in vitro batch culturing of a control (without biochanin A) and a biochanin A treatment. Following a 24-h incubation, gas production and the amounts of ammonia-nitrogen (NH(3)-N), volatile fatty acid (VFA), and amino acids were measured. Microbial population using 16S rRNA gene sequence. We found that the addition of biochanin A significantly increased microbial gas production; but had no effect on VFA production. Biochanin A supplementation also resulted in reduced microbial urease activity with half the maximal inhibitory concentration of 320 nM and also inhibited the degradation rates of total amino acids, valine, lysine, methionine and leucine by 18%, 56%, 37%, 13%, and 12%, respectively. This inhibition of urease activity and amino acid decomposition resulted in a significant reduction in the NH(3)-N concentration. High-throughput sequencing of the 16S rRNA sequence to monitor microbial composition showed that biochanin A significantly reduced the abundance of the proteolytic bacteria Prevotella and ureolytic bacteria Selenomonas, but increased the abundance of the lactic acid metabolizing bacteria Veillonella and Megasphaera. In conclusion, biochanin A reduced the production of ammonia by inhibiting proteolytic bacteria and their decomposition of urea and amino acids.