Abstract: Horses have always been animals used for companionship, work, transportation, and performance purposes over the history of humanity; there are different ways of managing horses, but studies on how horse welfare is influenced by different activities and managements are scanty. Understanding how the management, the environment, and the different uses of horses can affect the level of stress and well-being is important not only for people associated with horses. Three groups of horses with different management, environments, and activities were selected: (1) stabled horses ridden frequently, (2) horses that perform public order service under the Italian state police, and (3) free-ranging horses. Cortisol analysis was carried out on horsehair samples using liquid chromatography coupled to hybrid orbitrap high-resolution mass spectrometry (LC-HRMS/MS), a laboratory technique used for the first time to quantify horsehair cortisol. The selection of horses to be included in the three groups was carried out by including only subjects with positive welfare assessment in accordance with the horse welfare assessment protocol (AWIN). These analyses demonstrated that the cortisol levels detected in the horsehair of free-ranging animals were significantly higher compared to those detected in stabled and working horses. These results may have been a consequence of complex environmental, managerial, and behavioral factors, which should be worth further investigation

Abstract: Summary Domestic horses are equid members of the class Mammalia, order Perissodactyla, and family Equidae. Horses are obligate herbivores, with nutritional requirements as listed in a table. Adequate space is necessary for exercise, exploration, flight, sharing resources, play, and rolling. Company is essential for all horses, including stallions. Company provides opportunities for mutual grooming and play and allows horses to stand head-to-tail to remove flies. Unhandled horses may respond to humans as they would to predators, whereas handled horses' responses depend on their previous interactions with humans. Horses can suffer from several diseases as listed in another table. The best method of euthanasia of horses is usually sedation followed by either cranial shooting or the injection of an overdose of pentobarbitone into the jugular vein. Behavioural signs of distress can include increased locomotory activity, vigilance behaviours, neighing, snorting, pawing, nibbling walls and buckets, defaecation, rearing, kicking stable walls or doors, and high-stepping 'prancing'.