Our objective was to determine the effect of repeated use of implants on feedlot performance and carcass characteristics of Holstein cattle. Holstein steers (n = 128) weighing an average of 211 kg were blocked by weight and randomly assigned to 16 pens. At the start of the trial (d 0), pens were assigned to one of four treatments: 1) nonimplanted control (C); 2) implant on d 0, 112, and 224 (T3); 3) implant on d 112 and 224 (T2); and 4) implant on d 224 (T1). Component TE-S implants (120 mg of trenbolone acetate and 24 mg of estradiol per implant) were used for all treatments during the 291-d feeding period. Over the course of the study, T2 and T3 cattle had greater ADG and final weights than C and T1 cattle (P < 0.05). Steers were harvested at a commercial abattoir on d 291. Hot carcass weights of T3 steers were greater than those of C and T1 steers (P < 0.05). Dressing percentage, adjusted 12th-rib fat, percentage of kidney, pelvic, and heart fat, yield grade, and longissimus color were not different among treatments (P > or = 0.26). Longissimus muscle areas (LMA) of T2 and T3 carcasses were larger than LMA of C (P < 0.01). No USDA Select carcasses were produced from C cattle, whereas the percentage of Select carcasses from implanted cattle ranged from 10 to 18%. Skeletal maturity advanced (P < 0.05) progressively with each additional implant. Steaks from T3 carcasses had a higher percentage of protein than controls (P < 0.05) and were less tender than all other treatments (P < 0.05). Repeated administration of combination trenbolone acetate and estradiol implants increased ADG and resulted in heavier carcasses with larger LMA. Administration of three successive implants decreased tenderness of Holstein beef, and resulted in more advanced skeletal maturity scores.