Trials were conducted to evaluate effects of a fermentation extract of Aspergillus oryzae (AO) on milk production and composition, diet digestibility, and rectal temperature changes in lactating dairy cows. Treatments were incorporated as a top dressing at the morning feeding and consisted of control (90 g/d of ground sorghum) or AO (3 g of culture + 87 g of ground sorghum daily). Twenty-four mid-lactation Holstein cows were paired for production in Lactation Trial 1 (LT-1). In Lactation Trial 2 (LT-2), 46 cows (20 primiparous and 26 multiparous) in early lactation were used. Trials lasted 12 wk. In LT-1, AO supplementation increased milk yields only at 2 (P < .05) and 8 wk (P < .10) of treatment. Rectal temperatures were lower (P < .05) for cows fed AO for 4 of 10 readings made during summer. Supplementation of AO culture in LT-2 (early lactation cows) increased milk production and feed efficiency (P < .05). Inner ear temperatures tended to be lower (P < .11) for cows fed AO. Digestion trials, conducted at the end of lactation trials, used Cr2O3 as an indigestible marker. In Digestion Trial 1, digestibilities were not significantly (P > .10) affected by AO supplementation. However, in Digestion Trial 2, AO increased (P < .05) digestibilities of DM, OM, CP, NDF, and ADF. Length and number of meals were not affected (P > .10) by feeding AO. In summary, milk yields, efficiency of milk production, and nutrient digestibilities were higher for early lactation cows fed a high-concentrate diet supplemented with 3 g of AO/d. Mid-lactation cows fed a lower-energy diet were less responsive to AO than early lactation cows, though similar trends were shown.