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Two experiments were performed to evaluate the effects of bismuth subsalicylate (BSS) and calcium-ammonium nitrate (CAN) on in vitro ruminal fermentation, growth, apparent total tract digestibility of nutrients, liver mineral concentration, and carcass quality of beef cattle. In Exp. 1, four ruminally cannulated steers (520 ± 30 kg body weight [BW]) were used as donors to perform a batch culture and an in vitro organic matter digestibility (IVOMD) procedure. Treatments were arranged in a 2 × 2 factorial with factors being BSS (0 or 0.33% of substrate dry matter [DM]) and CAN (0 or 2.22% of substrate DM). In Exp. 2, 200 Angus-crossbred steers (385 ± 27 kg BW) were blocked by BW and allocated to 50 pens (4 steers/pen) in a randomized complete block design with a 2 × 2 + 1 factorial arrangement of treatments. Factors included BSS (0 or 0.33% of the diet DM) and nonprotein nitrogen (NPN) source (urea or encapsulated CAN [eCAN] included at 0.68% or 2.0% of the diet, respectively) with 0.28% ruminally available S (RAS). A low S diet was included as a positive control containing urea (0.68% of DM) and 0.14% RAS. For Exp. 1, data were analyzed using the MIXED procedure of SAS with the fixed effects of BSS, CAN, BSS × CAN, and the random effect of donor. For Exp. 2, the MIXED procedure of SAS was used for continuous variables and the GLIMMIX procedure for categorical data. For Exp. 1, no differences (P > 0.230) were observed for IVOMD. There was a tendency (P = 0.055) for an interaction regarding H2S production. Acetate:propionate increased (P = 0.003) with the addition of CAN. In Exp. 2, there was a NPN source effect (P = 0.032) where steers consuming urea had greater carcass-adjusted final shrunk BW than those consuming eCAN. Intake of DM (P < 0.001) and carcass-adjusted average daily gain (P = 0.024) were reduced by eCAN; however, it did not affect (P = 0.650) carcass-adjusted feed efficiency. Steers consuming urea had greater (P = 0.032) hot carcass weight, and a BSS × NPN interaction (P = 0.019) was observed on calculated yield grade. Apparent absorption of S decreased (P < 0.001) with the addition of BSS. Final liver Cu concentration was reduced (P = 0.042) by 58% in cattle fed BSS, indicating that BSS may decrease Cu absorption and storage in the liver. The results observed in this experiment indicate that BSS does not have negative effects on feedlot steer performance, whereas CAN may hinder performance of steers fed finishing diets., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)