In vitro method for determining the ruminal degradation rate of rapeseed meal protein using 15N isotope labelled ammonia nitrogen

An in vitro method based on observations of 14N and 15N isotope fluxes between ammonia N and non-ammonia (NAN) pools was established to study the ruminal degradation rate of rapeseed meal protein. Feed protein equal to 125 mg of N/l was incubated in the presence of rumen fluid, mineral buffer, and a carbohydrate mixture formulated to provide a constant supply of fermentable energy over the entire incubation period. The ammonia N was labelled with the 15N isotope, and the incubations were carried out for 0, 0.5, 1, 1.5, 2, 3, 4, 5, 6, 8, and 10 h. A model with six pools was used to estimate the rate of protein degradation to ammonia N and the rate of microbial N synthesis from ammonia N. The parameter values were adjusted based on the sizes of the ammonia 14N, ammonia 15N, 14NAN, and 15NAN pools observed at different time points over the incubation period. The rate of rapeseed meal N degradation was 0.06/h (0.028 standard deviation between runs), and the predicted effective protein degradability was 0.38 (0.122 standard deviation between runs). The current approach seemed appropriate for determining microbial N synthesis from ammonia N, but measurement of the direct incorporation of amino acids into microbial N may be required to adequately characterize the metabolic events involved in ruminal protein degradation.