Ruminal microbes of adult steers do not degrade extracellular L-citrulline and have a limited ability to metabolize extracellular L-glutamate

Kyler R Gilbreath, Gayan I Nawaratna, Tryon A Wickersham, M Carey Satterfield, Fuller W Bazer, Guoyao Wu

The microbial population within the rumen has long been considered to have the capability of extensively degrading all dietary AA. Results from our feeding trials revealed that this dogma is not correct. In vitro studies were conducted to test the hypothesis that certain AA undergo little degradation by ruminal microbes. Whole ruminal fluid (3 mL, containing microorganisms) from cannulated adult steers (~500 kg, n = 6) was incubated at 37 °C with 5 mM L-glutamine, L-glutamate, L-arginine, or L-citrulline for 0, 0.5, 1, and 2 h to determine time-dependent changes in the metabolism of these AA. Additional ruminal fluid was incubated with 0, 0.5, 2 or 5 mM L-glutamine, L-glutamate, L-arginine, or L-citrulline for 2 h to determine dose-dependent changes in their metabolism. An aliquot (50 μL) of the incubation solution was collected at the predetermined time points for AA analyses. There was extensive hydrolysis of L-glutamine into L-glutamate and ammonia, and L-arginine into L-ornithine, L-proline, and ammonia, but the near absence of catabolism of extracellular L-glutamate and no degradation of extracellular L-citrulline by ruminal microbes. There was little uptake of ¹⁴C-labeled L-glutamate and no detectable uptake of ¹⁴C-labeled L-citrulline by the cells. These results indicate, for the first time, that ruminal microbes of adult steers do not degrade extracellular L-citrulline and that metabolism of extracellular L-glutamate is negligible compared with their ability to extensively catabolize extracellular L-arginine and L-glutamine.