Our objective was to assess the influence of visual flow direction on physiological changes and symptoms elicited by cybersickness. Twelve healthy subjects (6 male and 6 female) were exposed to a 15-min virtual ride on a rollercoaster on two different days in a counterbalanced manner, such half of participants were facing forward during the first ride while another half was facing backward. Forehead skin conductance, heart rate and HRV parameters (SDRR, RMSSD) were collected as objective measures; subjective symptoms were assessed with the Motion Sickness Assessment Questioner immediately after exposure. We found that while nausea ratings at which participants terminated the experiment did not differ between forward/backward rides, the mean ride tolerance time was significantly longer during reverse ride compared to forward ride (6.1±0.4 vs 5.0±0.5 min, respectively, p = 0.01, η2 = 0.45). Analysis of HRV parameters revealed significant reduction in both RMSSD (p = 0.02, t = 2.62, η2 = 0.43) and SDRR (p = 0.01, t = 2.90, η2 = 0.45) in the forward ride; no such changes were found in the backward ride. We also found that amplitude of phasic changes in forehead skin conductance increased significantly in both ride directions. This increase however was significantly lower (p<0.05) in backward ride when compared to the forward ride. When assessed immediately post-ride, subjects reported significantly lower (p = 0.04) subjective symptom intensity after the reverse ride compared to the forward ride. We conclude that the direction of visual flow has a significant effect on the symptoms reported by the subjects and on the physiological changes during cybersickness.