Abstract Though much effort has been made to characterize the in vivo (IV) and in situ (IS) dry matter digestibility (DMD) of southeastern forages, there is a lack of information identifying methods that best represent the in vivo digestibility of these forages. Thus, the objective of our study was to compare IV and IS methods to determine which technique best represents the in vivo DMD of four bermudagrass (Cynodon dactylon [L.] Pers.) cultivars. In an IV experiment, ruminally fistulated heifers (n = 4) were randomly assigned to one of four bermudagrass cultivars (Coastal [COS], Russell [RUS], Tifton 44 [T44], or Tifton 85 [T85]) for four 30-d in vivo periods (21-d adaptation and 9-d collection) in a Latin square design. On d 28 of each period, rumen fluid was collected 4 h post-feeding for an accompanying IV experiment using a completely randomized design with a 4 × 2 factorial treatment structure. Factors in the IV experiment included bermudagrass cultivar (COS, RUS, T44, and T85) and digestibility method (Tilley and Terry [TT] or Goering and Van Soest [GVS]). On d 31 of in vivo periods 3 and 5, an accompanying IS experiment was conducted as a randomized complete block design with three treatment factors (in vivo diet [previous described], bermudagrass cultivar [previously described], and incubation timepoint [0, 0.25, 0.50, 0.75, 1, 2, 3, 4, 8, 12, 16, 20, 24, 48, 72, 96, 120, 144, and 168 h]). Correlations were computed between in vivo DM disappearance and each of the IV (TT and GVS) and IS (24, 48, and 72 h) methods. Regressions were calculated for each of the individual IV and IS measurements, and a stepwise regression was used to determine the predictive value of linear combinations of the measurements. None of the IV or IS DMD values were correlated with in vivo DMD (P ≥ 0.18). Individual IV and IS DMD values did not yield significant models through linear regression (P ≥ 0.18), though the best model (based on AIC) was TT IVDMD (r2 = 0.09; P = 0.26). Stepwise regression revealed that there was no linear combination of IV or IS DMD that improved prediction beyond single predictor models. While both IV and IS experiments remain viable tools to screen forages and make relative comparisons, results from this experiment are interpreted to mean that neither of these methods is suitable for prediction of in vivo performance.
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