Prenatal transportation stress (PNS) results in calves that are more temperamental and have greater circulating concentrations of cortisol compared to control calves. The objective of this experiment was to evaluate whether PNS alters the number of pituitary corticotrophs in mature Brahman cows. We hypothesized that the increased circulating cortisol concentrations previously characterized in this bovine model is associated with developmental changes in the anterior pituitary leading to an increased number of corticotrophs. Pregnant Brahman cows (n = 48) were transported in trailers for 2-hour periods at 60±5, 80±5, 100±5, 120±5, and 140±5 days of gestation. Non-transported pregnant cows (n = 48) were designated as the Control group. Control and PNS offspring heifers were managed together under the same environmental conditions. At approximately 5 yr of age, randomly selected non-pregnant cows (Control, n = 8; PNS, n = 6) were humanely harvested and the whole pituitaries were collected. Pituitaries were weighed, fixed in paraformaldehyde, serially dehydrated with graded ethanol, embedded in paraffin blocks, and cut into 5-μm sections. Immunohistochemistry was performed to detect cells expressing adrenocorticotropic hormone (ACTH) as a marker for corticotrophs. Three comparable sections from the midsagittal plane from each animal were processed using an ovine ACTH-specific antibody (Dr. A.F. Parlow, NIDDK). Five fields of view were analyzed per section (15 fields per animal). Anterior pituitary gland weight did not differ (P < 0.10) between groups (Control = 2.11 ± 0.12 g; PNS = 2.10 ± 0.15 g). The mean number of ACTH-positive cells between control (531±56 cells/section) and PNS cows (477±49 cells/section) also did not differ (P < 0.10). In conclusion, the number of pituitary corticotrophs in mature Brahman cows was not affected by prenatal transportation stress and is likely not a mechanism mediating the increased circulating cortisol concentrations seen in this bovine model of fetal programming.