1. Twelve grain mixtures, one lucerne (Medicago sativa) hay and one maize silage which had been used in mixed diets for which dietary nitrogen undegraded in the rumen (UDN) had been estimated with duodenally-cannulated cows, were studied. Total N in the feeds was fractionated into pool A (N soluble in borate-phosphate buffer), pool B (total N —(pool A+pool C)) and pool C (acid-detergent-insoluble N or residual N after 24 h incubation in protease solution). 2. N solubilization in protease solution containing 6.6 units/ml (substrate-saturating enzyme concentration) indicated the presence of subfractions in pool B, with different rates of solubilization. Such subfractions were not detectable from in situ, Dacron bag, estimates of N solubilization. 3. UDN was estimated using a dynamic mathematical model and rate-constants obtained from N solubilization in protease solution or in situ. For three grain mixtures tested using the protease technique the model predicted UDN values of 7, 10 and 12% compared with values of 47, 66 and 59% estimated in vivo. The full range of experimental feeds was tested using the in situ technique and UDN values predicted by the model were used to derive UDN values for twelve mixed diets. The latter values were significantly but not closely correlated with those determined in vivo (r20–41. P < 0.05). 4. An attempt was made to simulate rumen proteolysis in vitro by choosing a protease enzyme concentration (0.066 units/ml) providing a proteolytic activity similar to that of whole rumen fluid. The experimental samples of feed were subjected to simulated rumen proteolysis for 18 or 48 h to resemble the mean retention times in the rumen for grain mixtures and roughages respectively. The residual N at the end of incubation was considered as an estimate of UDN. The UDN values estimated from simulated rumen proteolysis and those determined in vivo for twelve mixed diets were in close agreement (r20.61, P < 0–01). 5. Simulated rumen proteolysis can serve as a simple, rapid and sensitive method to estimate UDN in a variety of feedstuffs.
Bibliographical noteFunding Information:
U. K. wishes to express his gratitude to the Ministry of Education & Culture, Government of India for a scholarship. Constructive criticisms from Peter H. Robinson during the course of this work is gratefully acknowledged. The technical assistance of Terry W. Strawn and partial funding from Agway, Inc., Syracuse and the Dairy Forage Cluster, USDA are sincerely appreciated.