Abstract Apparent trace metal absorption and tissue retention upon incremental levels of supplemental Zn, Cu, and Mn were studied in growing cattle. A total of 60 Holstein bulls were enrolled for the study and fed for 28 d a diet consisting of barley straw (15%), molasses (10%) and pelleted concentrate (75%; Zn = 78 ppm, Cu = 15 ppm, Mn = 91 ppm). Thereafter, 20 bulls were randomly selected and slaughtered for determination of baseline tissue trace metal composition. The remaining 40 bulls [body weight (BW) = 394 ± 20 kg] were then blocked based on BW in 8 blocks of 5 bulls each. Bulls within a block were randomly assigned one of the diets differing in supplemental levels of Zn, Cu, and Mn (all from sulfate form) with the following dietary contents of Zn, Cu and Mn, respectively: No supplement (38, 7, and 47 ppm), Dose 1 (61, 11, and 68 ppm), Dose 2 (78, 15 and 91 ppm), Dose 3 (123, 26, 136 ppm) or Dose 4 (195, 43, and 214 ppm). These diets were fed for 12 wk, and complete collection of feces and urine were performed on wk 4, 8 and 12 for assessment of trace metals balance and apparent absorption. Gastrointestinal tissues, whole organs (heart, liver, spleen, and kidney), bile, cervical vertebra, and tibia were collected at slaughterhouse from all bulls. All samples were dried, ground, and analyzed for minerals. Data were processed using dietary treatment as a continuous variable and also as categorial variable. Both approaches adjusted for the fixed effect of block (Proc Mixed, SAS 9.4). Whole-body Zn, Cu and Mn balance quadratically increased (P < 0.05) with supplementation, with a significantly higher values for Dose 4 compared with all other dietary treatment (Figure 1, P < 0.05). Dose 4 also led to a 2-fold increase in urinary Zn and Cu excretions (P < 0.05), and a 4-fold increase in urinary Mn excretion (P < 0.01). Although magnitudes of change were moderate (8 to 15%), increasing dietary Zn supply linearly increased spleen Zn content (P = 0.03), and tended to increase ruminal and hepatic Zn content (P < 0.10). Hepatic Cu linearly increased from 264 to 667 mg/kg DM with increasing dietary Cu supply (P < 0.01), whereas Cu concentration in bile only increased at Doses 4 and 5 (P < 0.01). Increasing Mn supply resulted in quadratic increase of ruminal and biliary Mn content from 340 to 1140 mg/kg DM, and from 0.32 to 1.38 mg/L, respectively. Small intestine and cecum Mn concentrations linearly and quadratically increased (about 2-fold increase, P < 0.05) with incremental supply of Mn. Homeostatic regulation mechanisms were apparently overwhelmed at the greatest Zn, Cu and Mn supplementation levels evaluated in this study.