Rhinoceros populations in Africa are under severe threat as a result of surging poaching rates and risk-mitigation strategies are continuously adapted in an attempt to ensure the survival of the species. This study compared faecal glucocorticoid metabolite (fGCM) levels of two age classes of limited free-ranging female white rhinos with fGCM levels of adult free-ranging female white rhinos. Subsequently, fGCM alterations in the limited free-ranging animals were monitored following routine dehorning as a measure of the animals’ short-term physiological stress response. Baseline fGCM levels differed significantly between tested groups, with both free-ranging and limited free-ranging adult animals showing significantly higher fGCM levels compared with limited free-ranging juvenile females. In contrast, baseline fGCM levels did not differ significantly between limited free-ranging and free-ranging adult individuals. Routine dehorning procedures resulted in a short-term stress response expressed by a significant increase in fGCM levels 48 h post-dehorning, with stress steroid levels returning to pre-dehorning concentrations 72 h after the procedure.