The consequences of stress on the behaviour of wild creek chub Semotilus atromaculatus outside the reproductive period were studied using a single intra-coelomic injection of cortisol, suspended in coconut butter, to experimentally raise plasma cortisol levels. Behaviour between cortisol-treated, sham-treated (injected with coconut butter) and control S. atromaculatus was compared in a mesocosm system, using a passive integrated transponder array, and in a natural stream system (excluding shams), using surgically implanted radio transmitters. While laboratory time-course studies revealed that the cortisol injection provided a physiologically relevant challenge, causing prolonged (c. 3 days) elevations of plasma cortisol similar to that achieved with a standardized chasing protocol, no differences in fine-scale movements were observed between cortisol-treated, sham-treated and control S. atromaculatus nor in the large-scale movements of cortisol-treated and control S. atromaculatus. Moreover, no differences were observed in diel activity patterns among treatments. Differential mortality, however, occurred starting 10 days after treatment where cortisol-treated S. atromaculatus exhibited nearly twice as many mortalities as shams and controls. These results suggest that, although the experimental manipulation of cortisol titres was sufficient to cause mortality in some individuals, there were compensatory mechanisms that maintained behaviours (i.e. including activity and movement) prior to death. This study is one of the first to use experimental cortisol implants outside a laboratory environment and during the non-reproductive period and yields insight into how wild animals respond to additional challenges (in this case elevated cortisol) using ecologically meaningful endpoints.