Measuring short-term stress in birds: Comparing different endpoints of the endocrine-immune interface

Stress is a collective term for certain conditions and sequences of physiological events enabling living organisms to overcome unpredictable and uncontrollable situations. The context-dependent nature, multidimensional course and large individual variability make stress responses difficult to measure. In avian species, a plethora of studies on short-term stress responses have been conducted by measuring the corticosteroid response to a standardized stress protocol. Here we aimed to test the viability of the leukocyte coping capacity (LCC), measuring oxygen radical production by leukocytes, to assess short-term stress in birds. We collected blood samples from captive house sparrows (Passer domesticus) in the two seasons of winter and spring, right after capture and 30 min thereafter. In order to assess the overall physiological stress response to a standardized stressor, i.e. handling and temporary constraint, we measured LCC and additionally combined it with measures of total circulating glucocorticoids (GCs) and oxidative stress. All three methodologies detected significant changes due to the stressor albeit they were not correlated with each other and revealed different information. There was no significant change in oxidative stress levels between the two time points although the amount of relative oxidative damage as well as the anti-oxidative capacity changed significantly. We observed a significant seasonal difference in GC stress response with no difference between sexes. On the contrary, LCC measures revealed with a high individual consistency, that individuals experienced a similar magnitude of stress in both seasons with a significant difference between sexes. Total GC-levels have to be interpreted with caution regarding the assessment of short-term stress reactions. We therefore suggest to supplementary combine classical approaches for measuring stress with the immunological tool of LCC. Our results reveal LCC as a strong and reliable tool to assess short-term stress in captive house sparrows and as promising for other bird species. Collectively the study highlights the necessity to incorporate a range of physiological systems and their endpoints to measure and to assess stress reactions effectively.