Cook, N. J. 2012. Review: Minimally invasive sampling media and the measurement of corticosteroids as biomarkers of stress in animals. Can. J. Anim. Sci. 92: 227-259. The measurement of corticosteroid hormones is commonly used as a biomarker of an animal's response to stress. The difficulties in obtaining blood samples and the recognition of the stressor effect of blood sampling are primary drivers for the use of minimally invasive sample media. In mammals these include saliva, feces, urine, hair, and milk. In birds, samples include excreta, feathers, egg yolk and albumin. In fish, corticosteroids have been measured in excreta and swim-water. Each of these sample media incorporate corticosteroids in accordance with the processes by which they are formed, and this in turn dictates the periods of adrenocortical activity that each sample type represents. Cortisol in saliva represents a time-frame of minutes, whereas the production of feces may be hours to days depending on the species. The longest time-integrations are for hair and feathers which could be over a period of many weeks. The sample media also determines the structural changes that may occur via processes of conjugation to glucuronides and sulfides, metabolic conversion via enzymatic action, and bacterial breakdown. Structural changes determine the optimum methodologies used to measure corticosteroid hormones. In most sample media, measurement of a specific corticosteroid is a requirement depending on the species, e.g., cortisol in most mammals, or corticosterone in birds. However, in samples involving products of excretion, methodologies that measure a broad range of structurally related compounds are probably optimal. The utility of minimally invasive sample media as biomarkers of stress responses depends on the degree to which the corticosteroid content of the sample represents adrenocortical activity. Commonly, this involves comparisons between corticosteroid concentrations in blood plasma with concentrations in the alternative sample media. This review focuses on the methodological and biological validation of corticosteroid measurements in minimally invasive samples as biomarkers of adrenocortical responses to stress.