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The associative effects (AE) between concentrate (C), peanut shell (P) and alfalfa (A) were investigated by means of an automated gas production (GP) system. The C, P and A were incubated alone or as 40 : 60 : 0, 40 : 45 : 15, 40 : 30 : 30, 40 : 15 : 45, 40 : 0 : 60 and 30 : 70 : 0, 30 : 55 : 15, 30 : 40 : 30, 30 : 25 : 45, 30 : 10 : 60, 30 : 0 : 70 mixtures where the C : roughage (R) ratios were 40 : 60 and 30 : 70. Samples (0.2000 ± 0.0010 g) of single feeds or mixtures were incubated for 96 h in individual bottles (100 ml) with 30 ml of buffered rumen fluid. GP parameters were analysed using a single exponential equation. After incubation, the residues were used to determine pH, dry matter digestibility (DMD), organic matter digestibility (OMD), volatile fatty acids (VFA) and ammonia nitrogen (NH3-N) of the incubation fluid, and their single factor AE indices (SFAEI) and multiple-factors AE indices (MFAEI) were determined. The results showed that group of 30 peanut shell had higher SFAEI of GP48 h, DMD, OMD and total volatile fatty acids (P < 0.05) and MFAEI (P < 0.05) than groups 60, 45 and 0 when C : R was 40 : 60. The group of 10 peanut shell showed higher SFAEI of GP48 h, DMD and OMD (P < 0.05) than groups 70, 55 and 40 and MFAEI (P < 0.01) when C : R was 30 : 70. It is concluded that optimal SFAEI and MFAEI were obtained when the C : P : A ratios were 40 : 30 : 30 and 30 : 10 : 60.

Multiple-factor associative effects of peanut shell combined with alfalfa and concentrate determined by in vitro gas production method