Abstract Measurement of corticosterone in various tissues has been used to investigate the stress response in reptile and amphibian species for decades. The tissue source from which corticosterone is measured reflects different periods of time and chronicity of stress levels in the subject, and different tissue collection methods differ in degree of invasiveness. Studies of corticosterone in keratinized tissues of reptiles, such as shed skin, are limited in number compared to hair and feather glucocorticoid studies in avian and mammalian species, but warrant continued research as they may reflect more different periods of time and chronicity of corticosterone levels than plasma or other tissues, and can be obtained in a minimally invasive manner. In this study, we measured corticosterone concentrations in both plasma and shed skin of Louisiana pine snakes (Pituophis ruthveni) that were all previously diagnosed with subclinical Cryptosporidium serpentis infection. We also tracked stressors experienced by different individuals to identify potential relationships between periods of increased stress and corticosterone levels in plasma and shed skin. There were no significant correlations between individual plasma and shed skin corticosterone levels, or between corticosterone levels in either tissue type and stressors experienced. This is the first study where corticosterone levels were measured in plasma and shed skin of P. ruthveni, and is the first known evaluation of plasma and shed skin corticosterone levels in a snake population previously testing positive for Cryptosporidium serpentis.