Objectives: Rescuers work in 24-h shifts and the demanding nature of the occupation requires adequate recovery between work shifts. The purpose of this study has been to find out what kind of changes in autonomic control may be seen during work shift and its recovery period in the case of rescuers. An additional interest has been to see if aerobic fitness is associated with recovery from work shifts. Material and Methods: Fourteen male rescuers (aged 34±9 years old) volunteered to participate in the study. Heart rate variability (HRV) was recorded for 96 h to study stress and recovery, from the beginning of a 24-h work shift to the beginning of the next shift. Aerobic fitness assessment included maximal oxygen uptake (VO2max) estimation with a submaximal bicycle ergometer test. Salivary cortisol samples were collected 0 min, 15 min, and 30 min after awakening on the 3 resting days. Results: Some HRV parameters showed enhanced autonomic control after the work shift. Stress percentage decreased from the working day to the 2nd rest day (p < 0.05). However, maximal oxygen uptake was not associated with enhanced parasympathetic cardiac control (p > 0.05). Cortisol awakening response was attenuated right after the work shift. Conclusions: The HRV findings show that recovery after a long work shift takes several days. Thus, rescuers should pay attention to sufficient recovery before the next work shift, and an integrated model of perceived and physiological measurements could be beneficial to assess cardiovascular strain among rescuers with long work shifts. Int J Occup Med Environ Health 2017;30(3):433–444