Chlortetracycline is an antibiotic that is used to increase weight gain, efficiency of gain, carcass grade, and conception rates. The objective of this experiment was to evaluate the effects of supplementation of 350 mg/d of chlortetracycline on ADG, G:F, BCS, thyroxine, and systemic glucose concentrations in yearling dairy heifers. Forty 12-mo-old Holstein heifers (initial BW = 363 +/- 21 kg) were housed in a free-stall barn with ad libitum access to feed and water for 104 d. A transition period was begun 14 d before the age of 12 mo to acclimate the heifers to the diet. The chlortetracycline-fed group (n = 20) consumed 328 +/- 8.2 mg of chlortetracycline/heifer daily. Measurements for BW, withers and hip heights, BCS, and health score were recorded weekly. Dry matter intake was measured daily. Blood was sampled every 4 d to determine plasma thyroxine and glucose concentrations and every 2 d to determine progesterone concentrations. Heifers were artificially inseminated on the first observed standing heat after 13 mo of age. There were no effects of chlortetracycline on ADG, G:F, withers and hip heights, BCS, blood glucose concentrations, peak progesterone concentrations, health, or conception rate. There was an interaction between treatment and time for chlortetracycline on serum thyroxine concentration. In the beginning of the experiment, serum thyroxine concentration was lower in heifers supplemented with chlortetracycline. There was no difference between treatments in thyroxine concentration at the end of the experiment. Chlortetracycline supplementation was not beneficial for yearling dairy heifers.