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This research paper addresses the hypothesis that substituting soybean meal with locally produced yeast protein from Cyberlindnera jadinii in barley-based concentrates for Norwegian Red (NR) dairy cows does not have adverse effects on milk fatty acid (FA) composition, rumen microbiota and sensory quality of milk. As soybeans also represent valuable protein sources for human consumption, alternative protein sources need to be investigated for animal feed. A total of 48 NR dairy cows were allocated into three feeding treatments, with the same basal diet of grass silage, but different concentrates. The concentrates were all based on barley, but 7% of the barley in the barley-concentrate (BAR; negative control) was replaced by either soybean meal (SBM; conventional control) or yeast microbial protein (YEA). The experiment lasted for a total of 10 weeks, including 2 weeks of adaptation with the soybean meal concentrate. Analysis of the feed revealed some differences in the FA composition of the YEA concentrate compared to the SBM and BAR concentrates. In milk, only two FAs (C17:1n-8cis9 and an unidentified isomer of C18:3) were significantly different between the YEA- and SBM-group, while six FAs differed between the BAR- and SBM-group. However, the amount of these FAs was low compared to the entire FA profile (<0.7 g/100 g). The experimental diets did not affect rumen microbiota nor the milk sensory quality. This study shows that C. jadinii can replace soybean meal as a protein source in concentrates (7% inclusion) for NR dairy cows fed a diet composed of grass silage and concentrates without any effects on rumen microbiota, and without compromising the FA composition or sensory quality of milk.