A series of experiments were conducted to determine nutritional strategies to improve the efficiency of N utilization in dairy cows when feeding co-products including wheat-based (W-DDGS) and corn-wheat blend distillers grains with solubles (B-DDGS), and dried whey permeate (DWP). In Experiment 1, the objective was to determine the effects of replacing canola meal (CM) as the major protein source with W-DDGS on ruminal fermentation, microbial protein production, omasal nutrient flow, and animal performance. Cows were fed either a standard barley silage-based total mixed ration containing CM as the major protein supplement (0% W-DDGS, control) or diets formulated to contain 10, 15 and 20% W-DDGS (dry matter [DM] basis), with W-DDGS replacing primarily CM. Diets were isonitrogenous (18.9% crude protein [CP]). Inclusion of W-DDGS to the diet did not negatively affect ruminal fermentation, microbial protein production, and omasal nutrient flow. However, there was a 0.7- to 2.4-kg increase in DM intake, and a 1.2- to 1.8-kg increase in milk yield after the addition of W-DDGS in place of CM. In Experiment 2, the objective was to delineate the effects of including either W-DDGS or B-DDGS dried distillers grains with solubles as the major protein source in low or high CP diets fed to dairy cows on ruminal function, microbial protein synthesis, omasal nutrient flows, urea-N recycling, and milk production. The treatment factors were type of distillers co-product (W-DDGS vs. B-DDGS) and dietary CP content (15.2 vs. 17.3%; DM basis). The B-DDGS was produced from a mixture of 15% wheat and 85% corn grain. All diets were formulated to contain 10% W-DDGS or B-DDGS on a DM basis. Feeding up to 10% of dietary DM as B-DDGS or W-DDGS as the major source of protein did not have negative effects on metabolizable protein (MP) supply and milk production in dairy cows. However, reducing dietary CP content from 17.3 to 15.2% decreased milk production. This response was attributed to an insufficient supply of ruminally degradable protein (RDP) that suppressed microbial nonammonia N (NAN) synthesis in the rumen, thus decreasing intestinal MP supply. In Experiment 3, the objective was to determine the effects of replacing barley or corn starch with lactose (as DWP) in diets containing 10% W-DDGS on ruminal function, omasal nutrient flow, and lactation performance. The treatment factors were source of starch (barley vs. corn) and dietary inclusion level of DWP (0 vs. 6%; DM basis) as a partial replacement for starch. Diets were isonitrogenous (18% CP) and contained 3 or 8% total sugar. The starch content of the low sugar diet was 24% compared to 20% for the high sugar diet. Dry matter intake, and milk and milk component yields did not differ with diet. However, partially replacing dietary corn or barley starch with sugar up-regulated ruminal acetate and propionate absorption, and reduced ruminal NH3-N concentration, but had no effect on ruminal pH, microbial protein synthesis, omasal nutrient flow and production in dairy cows. In summary, data presented in this thesis indicate that W-DDGS and B-DDGS can be included as the major source of protein in dairy cow diets without compromising ruminal function, nutrient supply and milk production in dairy cows. Feeding medium to low CP diets, and partial replacement of starch with sugar in diets containing W-DDGS and B-DDGS can improve N utilization efficiency in dairy cows. Additionally, an upregulation of facilitated transport of acetate and propionate across epithelial cells possibly prevents the occurrence of ruminal acidosis when lactose partially replaces starch in cow diets.