An endogenous adrenocorticotropic hormone (ACTH) concentration above the reference interval (RI) is commonly used as means for diagnosing equine pituitary pars intermedia dysfunction (PPID). Basal ACTH concentrations are highly dependent on photoperiod and RIs should be month- and location-specific. To date, no ACTH RIs have been specifically established for South Africa. This study aimed to determine geographically and seasonally relevant RIs for equine ACTH in the Gauteng province of South Africa. A longitudinal prospective study was conducted over twelve months to determine ACTH RIs for a representative population of healthy South African horses in the Gauteng province. Eighty clinically healthy horses under 12 years of age were recruited for monthly venous blood sample collection, from July 2019 to June 2020. ACTH was measured using a chemiluminescent assay. RIs were constructed for each month of the year. This South African population showed similar temporal changes in ACTH concentrations to those previously observed in other locations. Upper reference limits were at their lowest in early summer (21.4 pg/ml, 90% CI 20.8-21.7) with a pronounced increase in autumn (60.6 pg/ml, 90% CI 53.1-62.7), and tapered off in winter (22.3 pg/ml, 90% CI 19.9-23.2). The month-specific ACTH RIs generated in this study will improve the accuracy of diagnosis and monitoring of PPID in the local equine population. These results highlighted the previously recommended need for seasonal and location-specific RIs.
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