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Alpine forages are assumed to have specific effects on ruminal digestion when fed to cattle. These effects were investigated in an experiment from two perspectives, either by using such forages as a substrate for incubation or as feed for a rumen fluid donor cow. In total, six 24-h in vitro batch culture runs were performed. Rumen fluid was collected from a non-lactating donor cow after having grazed pastures at ∼2000 m above sea level for 2, 6 and 10 weeks. These "alpine runs" were compared with three lowland samplings from before and 2 and 6 weeks after the alpine grazing where a silage-concentrate mix was fed. In each run, nine replicates of four forages each were incubated. These forages differed in type and origin (alpine hay, lowland ryegrass hay, grass-maize silage mix, pure hemicellulose) as well as in the content of nutrients. Concentrations of phenolic compounds in the incubated forages were (g/kg dry matter (DM)): 20 (tannin proportion: 0.47), 8 (0.27), 15 (0.52) and 0 (0), respectively. Crude protein was highest in the silage mix and lowest with hemicellulose, whereas the opposite was the case for fiber. The total phenol contents (g/kg DM) for the high altitude and the lowland diet of the donor cow were 27 (tannins: 0.50 of phenols) and 12 (0.27), respectively. Independent of the origin of the rumen fluid, the incubation with alpine hay decreased (P < 0.05) bacterial counts, fermentation gas amount, volatile fatty acid (VFA) production as well as ammonia and methane concentrations in fermentation gas (the latter two being not lower when compared with hemicellulose). Alpine grazing of the cow in turn increased (P < 0.001) bacterial counts and, to a lesser extent, acetate proportion compared with lowland feeding. Further, alpine grazing decreased protozoal count (P < 0.05) and VFA production (P < 0.001) to a small extent, whereas methane remained widely unchanged. There were interactions (P < 0.05) between forage type incubated and feeding period of the donor cow in protozoal counts, acetate:propionate ratio, fermentation gas production and its content of methane, in vitro organic matter digestibility and metabolizable energy. Although increased phenolic compounds were the most consistent common property of the applied alpine forages, a clear attribution to certain effects was not possible in this study. As a further result, adaptation (long-term for donor cow, short term for 24 h incubations) appears to influence the expression of alpine forage effects in ruminal fermentation.