TeamGym is a popular form of gymnastics, including tumbling (Tu), trampette (Tr) and floor exercises (F) characterized by intensive practice placing high levels of stress on athletes. The aim of the study was to investigate athletes’ stress-related changes during TeamGym training and competition, considering hormonal and enzymatic responses (i.e., salivary cortisol and alpha-amylase). Ten (5 males and 5 females) TeamGym athletes (age: 22–28 y) were tested twice at the same time before training and competition; furthermore, for excluding circadian effect on hormonal and enzymatic responses, they were tested at the same time during a rest day. Alpha-amylase and cortisol were measured 15 min before the beginning of exercise, after each gymnastic equipment performance, and after thirty minutes from the end of the performance. Factorial ANOVA with repeated measures was used to verify differences between training and competition (p < 0.05). Competition elicited higher values of alpha-amylase than training (p ranging from 0.001 to 0.019) and rest (p ranging from 0.001 to 0.019). Cortisol showed no exercise induced increase, and its concentrations were higher prior to training compared to competition. TeamGym responses confirm other sports findings in stating that competition elicits higher stress response than training and suggest that salivary alpha-amylase is a more sensitive marker than cortisol to psychophysiological stress also in gymnastics intermittent performance.