The short-beaked common dolphin (Delphinus delphis) faces various anthropogenic threats, which can induce stress responses and lead to significant physiological adaptations to maintain homeostasis. The adrenal gland plays a crucial role as the primary endocrine organ, mediating the effects of both chronic and acute stressors through the production of mineralocorticoids and glucocorticoids. This study investigates stress-induced pathologies in common dolphins' adrenal glands using histological techniques and a novel proteomic approach. Postmortem examinations of 58 stranded common dolphins from the Irish coast (2017-2019) provided vital data, including body measurements, sexual maturity level, sex, decomposition state, nutritional status, and cause of death. Morphometric analyses included cortex-to-medulla (CM) ratio and the number of nuclei in the cortex (NNC). Multifactorial statistical analyses explored adrenal gland mass (AM), CM ratio, and NNC in relation to different stress types, sex, sexual maturity, body length, and nutritional status. Chronically stressed dolphins exhibited higher cortical mass and lower nuclei concentration, indicating possible hypertrophy. Additionally, a protein extraction protocol from the paraffin-wax-embedded adrenal glands was developed. The LC-MS/MS analysis performed, showed that the In-Gel digestion method yielded more identified proteins compared to the FASP method, possibly being more appropriate for the protein extraction from adrenal glands.
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