This study aimed to evaluate the effect of the level of crude glycerin (CG) on in vitro fermentation kinetics (0, 20, 40, 60, and 80 g/kg DM of sugar cane silage), on in vitro neutral detergent fiber (NDF) degradation (0, 30, 60, and 90 g/kg DM of sugar cane silage), and intake and digestibility of nutrients and nitrogen balance (0, 20, 55, 82, and 108 g/kg DM of sugar cane silage) in lambs. The in vitro trials were conducted in a completely randomized design with three repetitions. The in vivo trial was conducted in a Latin square design with five repetitions (5 × 5). For variables in which the F test was considered significant, the statistical interpretation of the effect of CG substitution levels was carried out through regression analyses. Kinetic parameters were not affected by CG inclusion. On in vitro NDF degradation, a significant effect of CG levels was observed on the potentially degradable fraction of NDF, the insoluble potentially degradable fraction of NDF, and the undegradable NDF fraction. The intake and digestibility of nutrients and nitrogen balance were not affected by CG inclusion. The CG levels change in vitro NDF degradability parameters; however, there were no changes in animal intake, digestibility, and nitrogen balance with the inclusion levels used.