Several studies have documented the occurrence and fate of trenbolone acetate (TBA) metabolites in soil and water. However, considerable uncertainty still exists with respect to TBA risk in agro-ecosystems because limited data are available to quantify excretion, transformation, and leaching processes. To address these uncertainties, we used experimental mesocosms and a mass balance approach to estimate the TBA metabolite leaching potential from manure excreted by implanted (40 mg TBA, 8 mg 17β-estradiol) beef cattle. Manure sample analysis indicates that over 113 days, a maximum of 9.3% (3,200 μg/animal unit [AU]) of the implant dose was excreted as 17α-trenbolone (17α-TBOH), and <1% was excreted as 17β-trenbolone (65 μg/AU) or trendione (3 μg/AU). While most (>97%) of the total excreted mass of 17α-TBOH transforms to uncharacterized products, 0.3-0.6% (100-220 μg/AU) of the implant dose accumulates on land surfaces and is available for subsequent transport. During rainfall or irrigation events, a maximum of 0.005-0.06% (1.6-22 μg/AU 17α-TBOH) or 0.005-0.012% (1.8-4 μg/AU 17α-TBOH) of the dose leached into runoff, respectively. Leaching potentials peak at 5-30 days postimplantation, suggesting that targeted timing of implantation and irrigation could minimize steroid leaching during rainfall and irrigation events.