The aim of this study was to examine the acute response to plasma and salivary cortisol and testosterone to three training protocols. Ten trained endurance athletes participated in three experimental trials, such as interval training (INT), tempo run (TEMP) and bodyweight-only circuit training (CIR), on separate days. Blood and saliva samples were collected pre- and 0, 15, 30 and 60 min post-exercise. Peak post-exercise salivary cortisol was higher than pre-exercise in all trials (P < 0.01). After INT, salivary cortisol remained elevated above pre-exercise than 60 min post-exercise. Salivary testosterone also increased post-exercise in all trials (P < 0.05). Plasma and salivary cortisol were correlated between individuals (r = 0.81, 0.73-0.88) and within individuals (r = 0.81, 0.73-0.87) (P < 0.01). Plasma and salivary testosterone was also correlated between (r = 0.57, 0.43-0.69) and within individuals (r = 0.60, 0.45-0.72), (P < 0.01). Peak cortisol and testosterone levels occurred simultaneously in plasma and saliva, but timing of post-exercise hormone peaks differed between trials and individuals. Further investigation is required to identify the mechanisms eliciting an increase in hormones in response to CIR. Furthermore, saliva is a valid alternative sampling technique for measurement of cortisol, although the complex, individual and situation dependent nature of the hormone response to acute exercise should be considered.