Degradability of forage proteins by in situ and in vitro enzymatic methods

The overall objective of these two studies was to evaluate the efficacy of using the proteolytic enzyme from Streptomyces griseus to estimate concentrations of ruminally degradable protein (RDP) in a wide array of forages. In the first study, alfalfa and prairie hays that previously had been evaluated in vivo for RDP were incubated in a replicated 3 x 3 factorial combination of enzyme concentrations (6.6, 0.66, and 0.066 activity units/ml of incubation medium) and incubation times (2, 4, and 48 h). Two treatment combinations (6.6 activity units for 4 h and 0.066 activity units for 48 h) yielded respective RDP estimates for alfalfa and prairie hay that were close to the known in vivo values. In the second study, 20 diverse forages were evaluated for RDP by using the in situ technique. These forages also were evaluated for RDP with the two enzyme concentrations identified in the first study, but incubation times were expanded to include 1, 2, 3, 4, and 5 h at the high concentration and 24, 30, 36, 42, 48, and 54 h at the low concentration. At the high enzyme concentration, r2 statistics from linear regressions of enzymatic estimates of RDP on corresponding estimates obtained by the in situ procedure were high (r2 > or = 0.898) at all incubation times; in addition, slopes (range = 0.88 to 1.00) and intercepts (range = -9.4 to 3.5%) approached unity and 0, respectively. At the lower enzyme concentration, r2 statistics were still good (> 0.81), but slopes (0.59 to 0.67) and intercepts (18.5 to 21.9%) for all incubation times did not meet the respective goals of unity and 0.