An analog of a bovine-appeasing substance (BAS) was previously demonstrated to have calming effects, and it could be an alternative to alleviate the stress caused by weaning. Thus, the objective of this study was to evaluate the effects of BAS administration at weaning on growth, stress, behavior, and response to vaccination of Nellore calves. Eighty-six Nellore calves (40 females and 46 males) were abruptly weaned and randomly assigned into 1 of 2 treatments: (1) saline solution (0.9% NaCl; n = 43) and (2) BAS (Secure Catte, IRSEA Group, Quartier Salignan, France; n = 43). The solutions were topically applied (5 mL/calf) to the nuchal skin area of each animal. On d 0, before treatment application, calves were vaccinated against infectious bovine rhinotracheitis (IBR), parainfluenza-3 (PI₃) virus, and bovine viral diarrhea virus types 1 and 2 (BVDV-1 and 2). Calves from each treatment were kept in different pastures for 15 d (time of BAS action) and then moved to a single pasture. Body weight (BW), blood samples, and temperament in the chute (entry score, chute score, and exit score) were collected on d 0, 3, 8, 15, 51, and 100, and behavior on pasture on d 1, 2, 4, 5, 6, 7, and 9. Calves assigned to BAS vs. Saline treatment tended to have greater BW on d 15 (p = 0.10), tended to have lower entry scores on d 8 and 51 (p = 0.10), and chute scores on d 8 (p = 0.07), and had lower exit scores on d 8 (p = 0.02). Calves assigned to BAS vs. Saline treatment also had greater time grazing on d 7 and 9 (p < 0.01), eating concentrate on d 2, 5, and 6 (p = 0.05), walking on d 1, 2, 5, and 9 (p < 0.01), standing and ruminating on d 2, 7 and 9 (p < 0.01), and playing on d 2, 4, 6, 7, and 9 (p < 0.01). Furthermore, they had lower time lying on 1 and 2 (p < 0.01), standing on d 5 and 9 (p < 0.01), and vocalizing on d 1 and 2 (p < 0.01). Calves assigned to BAS vs. Saline treatment had greater serum titter concentrations of PI₃ t on d 15 and 51 (p = 0.05) and BVDV-1 on d 51 (p = 0.02). However, they had lower serum concentrations of cortisol on d 3 (p = 0.03). BAS administration did not affect (p ≥ 0.12) the serum titer concentration of IBR and BVDV-2 titers or the plasma concentration of haptoglobin and ceruloplasmin. The BAS administration improved BW, reduced temperament and serum cortisol concentration, and improved behavior and response to vaccination.