Abstract The objectives of the study were to determine the effect of steroidal implants on growth performance, carcass characteristics, and estradiol-17β (E2) concentrations in the blood and longissimus muscle of Holstein steers fed a grain-based diet. Seventy Holstein steers (average initial BW = 275 ± 6.4 kg, 10 to 12 mo of age) were assigned to treatments: (i) implanted with 80 mg of trenbolone acetate (TBA) and 16 mg of E2 (Component TE-IS with Tylan; Elanco Animal Health, Greenfield, IN) at the start of the trial (day 0), and reimplanted with 120 mg of TBA and 24 mg of E2 (Component TE-S with Tylan; Elanco Animal Health) on day 84 of the experiment; or (ii) no implant. Implanted Holstein steers were heavier (P ≤ 0.01) than nonimplanted Holstein steers in the middle (day 84) and at the end of the experiment (day 186). Implanting Holstein steers increased (P < 0.01) average daily gain (ADG) and dry matter intake (DMI) without affecting gain-to-feed ratio compared with nonimplanted animals. Carcasses from implanted Holstein steers had greater (P < 0.01) hot carcass weight (HCW) and longissimus muscle (LM) area than carcasses from nonimplanted steers. Implanting did not affect (P ≥ 0.21) other carcass characteristics. There was an increase (P = 0.03) of 1.3 pg of E2/g of muscle in implanted Holstein steers compared with that from nonimplanted Holstein steers. There was an implant × day interaction (P < 0.01) in serum E2 concentrations. Serum E2 concentrations were not altered in nonimplanted Holstein steers, whereas E2 concentration increased (P < 0.01) after steers were implanted, regardless of implant characteristics. Serum E2 peaked at 28 days after the first implant and then rapidly declined after day 56. In summary, steroidal implants administered on days 0 and 84 increased DMI, ADG, HCW, and LM area in Holstein steers compared with nonimplanted steers due to increased serum E2 concentrations. However, these changes did not improve feed efficiency or other carcass characteristics.