One in vitro assay and one in vivo trial with ruminally cannulated Holstein steers were conducted to evaluate the effects of a dietary substitution of soybean meal by a urea and slow-release urea source of fermentation and degradation of diets for cattle. The experimental diets consisted of the total mixed rations defined as the control with soybean meal (SBM), U (urea), SRU (slow-release urea), and SRU+U+AA (0.42% + 0.42% + 1% amino acids methionine and lysine). The dietary substitution of SBM by U or SRU reduced (P < 0.05) the total gas production (V), microbial mass and degradation at 72 h incubation under the in vitro conditions, as well as the degradation rate (c) and the total volatile fatty acids (VFA) in the rumen of the steers; however, when the dietary substitution of SBM was by U+SRU+AA, those values did not decrease. In the steers, the dietary substitution of SBM by U and SRU reduced the ruminal degradation rate and the total VFA, and increased the ammonia N, but when SBM was substituted by U+SRU+AA in the diets, these changes were not observed. No advantage of SRU over U was found. The dietary substitution of SBM by U, SRU, U+SRU+AA did not modify the molar proportion of the VFA in the rumen nor were there changes in the nutrient digestion or excretion. Both the in vitro assay and the in vivo trial indicated that replacing SBM with U or SRU increases the ruminal ammonia N concentrations and reduces the degradation rate in the rumen, although those undesirable findings were not found when the SBM was replaced by U+SRU+AA. Therefore, it is feasible to replace the SBM with a combination of urea, slow-release urea, lysine and methionine in the diet for the ruminants.