Although various exercise regimens are commonly used as countermeasures to reduce the cardiovascular deconditioning induced by microgravity, the underlying mechanisms are not well understood. In this study we aimed to test whether lower limb resistance exercise with flywheel technology can prevent the fluid homeostasis alterations induced by 90-day head-down tilt bed-rest (HDT), and thus improve orthostatic tolerance. Total body water (TBW, measured by isotope dilution) and plasma volume (PV, calculated from the haemoglobin and the haematocrit) were measured in a control group (Co, n=9) and a countermeasure group (CM, n=9). Simultaneously, plasma atrial natriuretic peptide (ANP), renin (AR), and aldosterone (Aldo), as well as urinary anti-diuretic hormone (ADH), were measured. Orthostatic tolerance was evaluated with a 10 min +80 degrees tilt-test the first day of recovery. After HDT, both groups showed a comparable decrease in orthostatic tolerance [8.2 (0.9) min, Co; 8.0 (0.7) min, CM], PV [-4.7 (1.8)%, Co; -6.2 (2.5)%, CM, P<0.05] and TBW [-6.3 (5.4)%, Co; -3.7 (2.1)%, CM, P<0.05]. AR [97.4 (22.0)%, Co; 117.3 (26.4)%, CM] and Aldo [111.3 (58.4)%, Co; 100.6 (52.0)%, CM] increased significantly in both groups but the countermeasures produced no noticeable effects [data are expressed as mean (SE)]. The drop in ANP was also similar in both groups [-42.0 (15.2)%, Co; -51.1 (27.7)% for the CM]. Surprisingly, urinary ADH declined similarly in both groups during the basal data control period [-25.3 (5.2)%, Co; -26.1 (9.6)%, CM) and was sustained at this level during the 90-day HDT. These results show that, under the conditions described, the flywheel exercise device failed to improve characteristic manifestations of cardiovascular deconditioning and suggest that more frequent and powerful exercise, associated with another device (e.g. LBNP) might be a better countermeasure.