Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)The aims of this study were: a) to analyze the time of day effect on the aerobic capacity and time to exhaustion at aerobic capacity intensity (TE), and b) to analyze the physiological impact of handling and exercise testing during the light and dark periods, based on hematological parameters. Eighty rats were randomly divided into two control groups (C12 and C20) and two exercise groups (E12 and E20), assessed at 12:00 h (C12 and E12) or 20:00 h (C20 and E20). The lactate minimum intensity (LMi) was measured and after 48 h the exercise groups were subjected to a bout of swimming until exhaustion at LMi (TE). The TE was 1.30 +/- 0.51 h for the E12 group and 1.81 +/- 0.77 h for the E20 group (p = 0.03). The time of day effect was significant for all white blood cell counts (12:00 h > 20:00 h). Chronic handling and performing exercise tests at 12:00 h (light period) resulted in an increased WBC counts and decreased exercise tolerance. The favorable time of day for aerobic capacity and performance assessment and hematological parameters was at 20:00 h (dark period), which is associated with the wakefulness period of the assessed animals.1004427434Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP [Proc. 2010/13377-7, 2011/13226-1]