Physiological stress responses have been associated with lesser pregnancy rates in beef cattle. Alternative cattle restraint methods have been developed to reduce this negative effect. The objective was to determine if there was a difference in pregnancy rate as a result of fixed-time artificial insemination (FTAI) in Bos taurus females when using a breeding box compared with a squeeze chute for restraining cows when conducting FTAI. There were FTAI treatment regimens imposed on cows and heifers at two separate locations (Virginia Tech University, VT; and R.A. Brown Ranch, RAB) with the FTAI being conducted while animals were restrained in a squeeze chute (n = 169) or a breeding box (n = 162). There was imposing of a 7-day Co-Sync + CIDR pre-breeding treatment regimen. Females were randomly assigned to treatments just prior to breeding. Immediately following breeding, blood was collected using coccygeal venipuncture procedures for assessment of plasma cortisol concentration. Chute and exit scores (1–5) were determined at the time of FTAI. Pregnancy rate, cortisol, chute score, and exit score were analyzed. There was no difference in pregnancy rate between treatments (57 ± 26% and 69 ± 26 %, respectively P = 0.59).There were differences in chute (1.52 ± 0.05 and 1.27 ± 0.05, respectively P < 0.05) and exit (1.26 ± 0.04 and 1.08 ± 0.04, respectively P < 0.05) scores between the two treatment groups. Results from the present study are not conclusive in ascertaining effects of the two types of restraint during FTAI on reproductive efficiency.