Calming supplements are common in the equine industry. This study tested the hypothesis that Phytozen EQ, a blend of citrus botanical oils, magnesium, and yeast would reduce startle response as well as reduce behavioral and physiological signs of stress in young (1.5–6 years of age) horses (n = 14) when tied in isolation and when trailered in isolation. During the 59-day trial, horses were assigned to either the control (CON; n = 7) or treatment (PZEN; n = 7) group that received 56 g of Phytozen EQ daily. Horses underwent a 10-minute isolation test on d 30 and a 15 minute individual trailering test on day 52 or 55. For both tests, blood samples were obtained pre, immediately after, and 1-hour post for analysis of plasma cortisol concentrations, which were analyzed by repeated measures ANOVA. On day 59, horses underwent a startle test, for which time to travel 3 m and total distance traveled were recorded. These data were analyzed using a T-test. During trailering, PZEN horses tended to have lower overall geometric mean (lower, upper 95% confidence interval) cortisol concentrations than CON (81 [67, 98] vs. 61 [48, 78] ng/mL; P = .071). For the startle test, PZEN horses tended to have longer geometric mean times to travel 3 m than CON horses (1.35 [0.39, 4.70] vs. 0.26 [0.07, 0.91 seconds, P = 0.064). Other data points were not different between treatments (P > .1). It is possible that this dietary supplement could have beneficial calming effects on horses undergoing trailering or in novel situations.