In seasonal breeders, periods of reproductive activity often coincide with high levels of glucocorticoids. We studied seven male and seven female African lesser bushbabies (Galago moholi A. Smith, 1836) over two mating periods via noninvasive faecal hormone metabolite monitoring to investigate the relationship between reproductive and adrenocortical hormone activity. We used linear mixed-effect models to investigate the effect of physiological (endocrine) variables on faecal glucocorticoid metabolite concentrations. Our results indicate faecal androgen (males) and progestagen metabolite (females) concentrations as the variables best able to explain variability in faecal glucocorticoid metabolite concentrations. However, the models explained only a fraction (26% and 12%, respectively) of the observed variability and graphical analysis suggests a biologically relevant difference in faecal glucocorticoid metabolite concentrations between captive and free-ranging animals during nonreproductive periods. Thus, captivity may have affected glucocorticoid output in our focal animals, potentially weakening the expected relationship between reproductive activity and faecal glucocorticoid metabolite variability. Due to the ease of faecal and observational sample collection, a large number of studies monitoring adrenocortical activity in wildlife are conducted using only captive settings, with inferences unquestioned when applied to free-ranging scenarios. Our study cautions against this practice, as particular housing or management conditions may influence the pattern of adrenocortical activity.