Because of the Ca dependency of the calpains, oral supplementation of vitamin D3 (VITD) can increase the Ca content of muscle to activate the calpains and improve tenderness. Feedlot steers (n = 142) were arranged in a 4 x 3 factorial arrangement consisting of four levels of VITD (0, 0.5, 1, and 5 million IU/[steer x d]) for eight consecutive days antemortem using three biological types (Bos indicus, Bos taurus-Continental, and Bos taurus-English). Feedlot performance factors of ADG, DMI, and G:F were measured, and carcass quality, yield, and color data were collected. Plasma Ca and P concentrations were measured during d 4 to 6 of supplementation and at exsanguination, and carcass pH and temperature were measured in the LM at 3 and 24 h postmortem. Vitamin D3 treatment at 5 million IU/(steer x d) decreased ADG (P < 0.05) over the supplementation and feed intake for the last 2 d of feeding compared with untreated control steers. Likewise, G:F was decreased (P = 0.03) in steers supplemented with 5 million IU/d compared with controls. Overall, there was a linear decrease (P < 0.01) in ADG and G:F as a result of VITD supplementation. Plasma concentrations of Ca and P were increased (P < 0.05) by VITD concentrations of 1 and 5 million IU/(steer x d). All VITD treatments increased (P < 0.05) LM temperature at 3 h postmortem and pH at 24 h postmortem. Vitamin D3 treatments did not affect (P = 0.07) any other carcass measurements, including USDA yield and quality grade; thus, any improvements in meat tenderness as a result of VITD supplementation can be made without adversely affecting economically important carcass factors. Biological type of cattle did not interact with VITD treatment for any carcass or feedlot performance trait. Although feeding 5 million IU/(steer x d) of VITD for eight consecutive days had negative effects on performance, supplementing VITD at 0.5 million IU/ (steer x d) did not significantly alter feedlot performance.