An evaluation of the validity of an in vitro and an in situ/in vitro procedure for assessing protein digestibility of blood meal, feather meal and a rumen-protected lysine prototype

Abstract In vitro procedures are commonly used to estimate rumen protein degradability and protein digestibility of feed ingredients. However, it is unclear how well these assays correlate to in vivo data. The objectives of this work were to compare postruminal protein availability estimates from one in vitro procedure and one in situ/in vitro procedure with in vivo observations for blood meal (BM), feather meal (FM), and a rumen-protected lysine prototype (RP-Lys). The FM and BM used for this experiment were subsamples of material assessed in vivo by an isotope-based method and the RP-Lys subsamples were of a prototype tested in two in vivo trials: a lactation trial and by plasma appearance. Subsamples of the BM (n = 14) and the FM (n = 22) were sent to each of three different laboratories for in vitro or in situ/in vitro analysis of crude protein (CP) and determination of rumen undegraded protein (RUP) and digested RUP (dRUP). Subsamples of the RP-Lys (n = 5) were sent to one laboratory for in vitro analysis of CP, RUP, and dRUP. Two diets containing BM or FM were assessed using the Cornell Net Carbohydrate and Protein System (CNCPS) v6.55 with ingredient inputs derived from either the CNCPS feed library, the isotope dilution method, or an average of the in vitro results from the three laboratories to determine how much the differences among estimates affected ingredient values. In vitro dRUP estimates for BM from one laboratory closely matched those determined in vivo (66.7% vs. 61.2%, respectively), but no in vitro estimates for FM matched the in vivo values. Surprisingly, there were significant differences in protein digestibility estimates from the modified three-step procedure across the two laboratories for BM (P < 0.0001) and for FM (P < 0.0001) indicating significant variation among laboratories in application of the method. Within all laboratories, BM estimates were reported in a narrow range (CV values of 2.6 or less). However, when testing multiple samples of FM or the RP-Lys prototype, CV values within a laboratory ranged up to 11 and 34, respectively. For the RP-Lys, dRUP estimates from the in vitro method were roughly half of that determined by the in vivo methods suggesting poor concordance between the in vitro and in vivo procedures for this ingredient. The inconsistencies within and among laboratories accompanied with dissimilarities to in vivo data is problematic for application in nutrition models. Additional refinement to the in vitro