This study measured cortisol concentrations in hair samples from beef cattle in Denmark to assess chronic stress. Hair was collected from 24 farms across the country and cortisol was extracted and analyzed. The results showed that hair cortisol: 1) varied geographically and between breeds, ages, and housing/management conditions; 2) was affected by factors like leg cleanliness and physiological state; and 3) while complex, may provide a measure of chronic stress in cattle. Further investigation of the identified factors is needed.