Objectives To compare acute cognitive effects following bouts of standing (STAND), cycling (CYCLE) and walking (WALK) to a sit-only (SIT) condition. Design Randomized cross-over full-factorial study. Methods Nine overweight (BMI=29±3kg/m2) adults (30±15years; 7 females, 2 males) completed four conditions (SIT, STAND, WALK and CYCLE) across a 6h period with a 7days washout period between conditions. SIT consisted of uninterrupted sitting. Experimental conditions included intermittent bouts of standing (STAND), cycling (CYCLE) and walking (WALK). A cognitive performance battery (Cogstate) was completed twice in a seated position following bouts of standing and light-intensity physical activity. Mixed-effects models compared between-condition differences in standardized score (z-score), accuracy (%), and speed (log10ms). Results Cognitive performance z-score and accuracy measures were higher during STAND, CYCLE and WALK (P <0.05) conditions compared to the SIT condition. CYCLE was better than other experimental conditions. Conclusions Compared to uninterrupted sitting, short bouts of standing or light-intensity cycling and walking may improve acute cognitive performance.