May Salivary Chromogranin A Act as a Physiological Index of Stress in Transported Donkeys? A Pilot Study

SIMPLE SUMMARY: Transportation is recognized as a stressful animal husbandry procedure, inducing both short term and prolonged effects on welfare. Donkeys are transported for several purposes; therefore, validating a reliable and non-invasive stress indicator is pivotal. This study aimed to investigate whether salivary chromogranin A (CgA) concentration, known as an index of stress in humans and pigs, could represent a novel index of transportation-induced stress in donkeys. The research involved the measurement of salivary CgA in 19 donkeys, 15 min before and 15 min after two transportations. The transportation, which took place on two consecutive days, followed the routine procedures of the farm. The analysis of salivary CgA was carried out by an enzyme-linked immunosorbent assay test. Results showed that CgA salivary levels significantly decreased after both transportations. The physiological mechanisms underlying this result may be related to catestatin acting as an inhibitor of catecholamine release; however, due to the limited number of subjects involved, this hypothesis requires further investigation. ABSTRACT: Road transport is known to be a stressful animal husbandry procedure as it induces the activation of two main physiological stress-related pathways: the hypothalamic-pituitary-adrenal cortex axis and the sympathetic-adrenal medulla axis. This preliminary study aimed to investigate whether salivary chromogranin A (CgA) concentration, known as a biomarker of the sympathetic activity system during psychological stress, may represent a novel physiological index of transportation-induced stress in donkeys. Nineteen Romagnolo donkeys, raised in groups on paddocks, were subject to two transportations, following the farm’s routine procedures, for a mean duration of 64 min each on two consecutive days. Salivary samples were gently collected 15 min before and 15 min after each transportation. Salivary CgA was measured by a commercially available enzyme-linked immunosorbent assay test. Results showed that CgA salivary levels significantly decreased after both transportations. The physiological mechanisms underlying this result may be related to catestatin activity, a bioactive product of the proteolytic cleavage of CgA, that acts as an inhibitor of catecholamine release. This hypothesis requires further investigation, particularly considering the limited number of subjects involved in this preliminary study. The identification of a reliable and non-invasive stress-marker would represent a useful tool for improving farm animals’ welfare in transport conditions.