Calf rope-and-tie is a competitive rodeo event that has drawn criticism due to concerns around the potential for stress and injury of calves. While previous studies have utilised behavioural indicators to assess the welfare of rodeo calves in the arena, there are no published studies on rodeo calf behaviour during confinement in the chute prior to release into the arena. We analysed video footage of calves in the chute prior to release during two rodeo events. The footage was categorised into short duration (SD, 28–51 s, n = 17) and long duration (LD, 52–166 s, n = 14) based on the total length of the video clip of each calf in the chute. To enable comparison, the final 28 s from both SD and LD categories before calf release were reviewed, and an ethogram was developed to assess the frequency of observable calf behaviours. All calves expressed at least one behaviour indicative of a negative mental state, with ear and head movements being the most frequent. Behaviours observed in SD and LD clips were strongly correlated (Spearman’s rank correlation coefficient 0.875, p < 0.001). Longer confinement was associated with increased frequency of “red flag” behaviours indicating fear and/or stress (chi-square = 10.48, p = 0.0149), including mouth opening, tongue protrusion, rearing and escape attempts. We conclude that calves used in roping events experience fear and stress while held in the chute, especially during longer periods of confinement.