The behaviour of oestrous mares is well-known in horse breeding. However, alterations in their physiological and behavioural indices during the whole oestrous cycle are scarcely known. The objective of the study was to analyse changes in cardiac activity variables, rectal and superficial temperatures, behaviour towards humans and conspecifics, and the time of standing and locomotor activity in mares during their oestrous cycle. Fifteen adult mares in oestrus were examined in the morning and evening (six successive days) and in dioestrus (five days—once every third day). The oscillation of physiological and behavioural variables accompanies changes in mares’ sexual behaviour. Most physiological variables studied in oestrus indicate the elevated activity of the adrenergic nervous system and, opposite to that, both behaviour towards humans and conspecifics and the time of standing relate to a relaxed state. The end of oestrus, manifested by a rapid decrease in most of the physiological variables studied, is followed by changes of behavioural variables at the beginning of dioestrus. The time of locomotor activity arises at the end of oestrus. The outcomes may contribute to the knowledge of, among others, mare owners who evaluate the oestrus by mares’ sexual behaviours without regarding other rhythmically changing variables.