Effects of inclusion of purple prairie clover (Dalea purpurea Vent.) with native cool-season grasses on in vitro fermentation and in situ digestibility of mixed forages
Authors: Peng Kai , Gresham Gemma L. , McAllister Tim A. , Xu Zhongjun , Iwaasa Alan , Schellenberg Mike , Chaves Alex V. , Wang Yuxi 
Published: February 2020

Abstract
Background
Incorporation of legume species into native North American pastures is considered an effective method to increase native pasture productivity and improve the nutritive value of forage. This study evaluated the effects of inclusion of purple prairie clover (PPC, Dalea purpurea Vent.), a native legume forage, with native cool-season grasses on the in vitro fermentation and in situ digestibility of mixed forages.

Methods
Whole plant PPC and mixtures of cool-season grasses were harvested when the PPC reached the vegetative (VEG), full flower (FL) and seedpod (SP) stages, and were combined in ratios (DM basis) of 0:100, 25:75, 50:50, 75:25 and 100:0 at each maturity. In vitro ruminal incubations using these mixtures were conducted for 48 h to determine gas production (GP), in vitro DM disappearance (IVDMD), total volatile fatty acids (VFA) and ammonia-N production. Mixtures of forages harvested when the PPC reached the FL stage and 50:50 mixture of forages harvested at VEG, FL and SP stages were incubated in the rumen of three heifers for 0, 2, 6, 12, 24, 48, 72 and 96 h to determine in situ degradabilities of DM, neutral detergent fibre (aNDF) and crude protein (CP).

Results
Contents of aNDF and ADF increased (P < 0.01), while CP decreased (P < 0.001) as PPC matured. Concentrations of extractable condensed tannins in PPC ranked as FL > VEG > SP (P < 0.05). Regardless of PPC proportions in the mixture, GP decreased (P < 0.05) with increasing PPC maturity. Increasing PPC proportions linearly increased (P < 0.001) GP, IVDMD and total VFA at VEG, but linearly decreased (P < 0.001) them at SP. Irrespective of PPC maturity, ammonia-N production linearly increased (P < 0.01) with increasing proportions of PPC and the concentration was higher (P < 0.05) at VEG than at FL and SP stages. Increasing proportion of PPC at either maturity linearly increased (P < 0.001) molar percentage of acetate (A) and branched-chain VFA, but linearly decreased (P < 0.001) molar percentage of propionate (P), resulting in a linearly increase (P < 0.001) in the A:P ratio. Increasing FL PPC in the mixture linearly and quadratically (P < 0.01) increased a (soluble fraction), but linearly and quadratically decreased (P < 0.01) b (potentially degradable fraction) for DM and aNDF, resulting in linear (P < 0.05) and quadratic (P < 0.01) increases in DM and aNDF maximum potential degradabilities (a + b). Effective degradabilities of DM and aNDF were also linearly and quadratically increased (P < 0.05), and CP was quadratically increased (P < 0.05) with increasing FL PPC, with the greatest effective degradability being observed with ratios between 50:50 and 75:25. Ruminal maximum potential degradabilities of DM and aNDF decreased (P < 0.001) as the forage matured. Effective degradability of DM ranked as VEG > FL > SP (P < 0.001), whereas the effective degradability of aNDF was similar between VEG and FL and both were greater (P < 0.01) than SP.

Conclusions
Inclusion of vegetative PPC in a mixed forage diet resulted in the greatest digestibility and incorporation of PPC before seedpod stage with native grasses had a positive effect on ruminal fermentation. Effects of PPC on ruminal digestion depend on both the stage of maturity and its proportion in mixed legume-grass pastures. Pastures containing 50% of PPC in full flower stage would likely provide the greatest quality diet to grazing ruminants subject to potential animal selectivity.