“…To assess the occurrence of stress in horses, the concentration of pituitary–adrenal axis hormones determined in saliva is increasingly used [ 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 ]. Non-invasive methods for assessment of stress response increasing, e.g., the level of free transcortin-unbound cortisol [ 9 , 10 , 11 ], are used in other animal species as well [ 12 , 13 , 14 , 15 , 16 ]. Such biological material is sampled with no additional pain, stress, or discomfort and the material can be collected by non-veterinarians [ 9 , 11 , 17 ].…”