Background: Few studies have been performed to identify objective indicators for the selection of therapeutic donkeys or to assess their welfare during animal-assisted interventions (AAIs) Objective: This study aimed to evaluate the response to the ethological test and the modifications of physiological parameters in donkeys subjected to AAI sessions. Methods: Thirteen donkeys were subjected to a behavioral evaluation during an AAI session. Heart rate, heart rate variability, and root mean square of successive difference values were detected. Results: Statistically significant changes in the tested parameters were observed during AAI sessions. Conclusions: In donkeys, there was a neurovegetative involvement during AAI sessions. Our data give a contribution to the evaluation of donkey welfare during AAIs.

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The values for basal serum cortisol concentrations of horses are available in many studies. However, there are limited data about serum cortisol in donkeys. The present study aimed to determine the baseline values for serum cortisol, to evaluate the influence of age and pregnancy on its levels, and to correlate its values with leucocyte profile, serum cholesterol, and triglycerides. Serum samples were collected from 97 healthy donkeys. Cortisol was analyzed by chemo-luminescent assay. The median and the 2.5th and 97.5th percentiles of serum cortisol measured and calculated in all donkeys were 5.64, 3.40, and 10.54 µg/dl, respectively. Females (n.91) were divided into three groups: Group A (young), Group B (adult), and Group C (pregnant at the 9th–11th months). The effect of age and physiological status was investigated by the Mann–Whitney test. Group C showed significantly higher levels than Group B (p < 0.05). Significant correlations were found in Group B with monocytes (r = 0.37, p < 0.01) and triglycerides (r = 0.30, p < 0.05), and in Group C with monocytes (r = 0.79, p < 0.01), basophils (r = 0.6, p < 0.05), and neutrophil/lymphocyte ratio (r = −0.63, p < 0.05). Higher cortisol values related to late pregnancy are also found in this species. These preliminary results provide evidence for a relationship between cortisol and the immune system as well as cortisol and lipid metabolism modulated by age and pregnancy when parameters are within normal values.