The objective of this study was to determine the contribution of the scrotum, testes, and the testicular artery to scrotal/testicular thermoregulation in bulls at two ambient temperatures. Crossbred beef bulls, 1.5 years of age, were placed in controlled environment chambers at ambient temperatures of 15 degrees C (n = 5) or 25 degrees C (n = 6). The distal lateral aspects and entire ventral part of the scrotum was incised under caudal epidural anaesthesia (xylazine, 0.07 mg kg-1. Both testes were withdrawn from the scrotum and then replaced and maintained by clamping the scrotal incisions with towel clamps. One testis was randomly chosen to be the exposed testis and was withdrawn prior to temperature measurements. Surface and internal temperature were measured with infrared thermography and needle thermocouples, respectively. Temperature gradients (degree C; difference in temperature from top to bottom at 15 and at 25 degrees C) were: scrotal surface (with testis), 1.5 and 1.3; scrotal surface (without testis), 2.1 and 1.6; surface of exposed testis, -0.6 and 0.0; sub-tunic of exposed testis, -2.2 and -0.6; intratesticular (covered testis), 0.0 and 0.4; and intratesticular (exposed testis), -1.3 and 0.4. The scrotum markedly affects testicular temperature but the testes have limited influence on scrotal surface temperature. The bovine scrotum and testes have opposing temperature gradients that complement one another, resulting in a relatively uniform intratesticular temperature. These temperature gradients are attributed in part to the testicular artery, which goes from the top of the testis to the bottom, divides into several branches and ramifies dorsally and laterally before entering the testicular parenchyma. Intra-arterial temperatures (measured with needle thermocouples) were lower (P < 0.05) where the artery entered the testis than at both the bottom and top of the testis for both the covered (31.7, 33.4 and 34.3 degrees C) and exposed testis (29.6, 32.0 and 32.5 degrees C) at an ambient temperature of 15 degrees C. Temperature differences were similar, but less pronounced, at 25 degrees C (covered testis, 34.8, 36.3 and 36.5 degrees C; exposed testis, 32.4, 33.5, 33.9 degrees C). Results supported the hypothesis that blood within the testicular artery has a similar temperature at the top of the testis (just ventral to the testicular vascular cone) compared with the bottom, but subsequently cools before entering the testicular parenchyma.