Abstract: Little research has been done to measure reactivity objectively in therapeutic riding horses (TRH). As individual reactivity and chronic stress could be assessed by exposing animals to acute, novel stressors, the authors of this work aimed at comparing reactions of TRHs and jumping horses (JH) to two challenges. Four TRHs and four JHs were exposed to a restraint covering their head with a hood for 1 h and to a startling stimulus (a 40 cm long, red and white synthetic holiday garland shaken with a rustling noise inside the box). Heart rate (HR) and heart rate variability (HRV) were recorded continuously and telemetrically, the reaction was video-recorded and analysed with a software for behavioural analysis. Blood samples were collected before and after each challenge to determine lymphocyte proliferation and other biochemical parameters. Horses spent most of the time immobile, during the challenges (p < 0.05). TRHs had a significantly higher average basal HR than JH (p < 0.05), probably due to their better condition. HR varied among different behaviours during the restraint (p < 0.05): the average HR during “pawing” was higher than during other behaviours (p < 0.005). A significant decrease in the proliferation of lymphocytes in samples taken after the removal of the hood (p < 0.05) was found, while the other stress related parameters did not vary significantly after the challenges. The authors conclude that TRHs did not react less than JHs to the new stimuli and this should be taken into consideration while planning their daily work and management.