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Introduction While sleep duration is known to affect next-day cognitive performance and alertness, largely in a dose-response manner, the effects of disrupted sleep (where one is awoken multiple times overnight, common in military settings) are much less understood. Therefore, we examined the effects of experimentally disrupted sleep on morning cognitive performance and alertness. Methods We tested 34 healthy participants (12 men, 22 women, 28.1±3.9 years; mean±SD) who slept for 8-hours time-in-bed on three consecutive nights with polysomnography in a controlled sleep lab. The final two nights were randomized and counterbalanced between an undisrupted and a disrupted sleep condition. On the disrupted sleep night, participants were awoken by auditory tones for a 5–10 min period every hour. The following morning, participants completed a cognitive test battery that included Karolinska Sleepiness Scale (KSS), 10-min psychomotor vigilance task (PVT), addition calculations (ADD), go/no-go (GNG), task switching (TS), and working memory (WM). Mixed effects models were used to test factors: condition (undisrupted vs. disrupted), condition-order, and their interaction. Results Significant (p<0.05) effects of condition (i.e., disrupted sleep caused worse performance) were found for PVT reaction time (RT), GNG RT, TS RT, WM percent correct, and KSS alertness ratings. Condition was not significant for number or percent correct on ADD, GNG, and TS. Condition-order was significant for TS percent correct, and significant interactions were found for ADD number correct and TS RT. Conclusion One night of sleep disruption caused significant negative effects on morning subjective alertness and on several, but not all, cognitive performance domains tested, including RT and WM. Condition-order and interaction effects were also found, indicating that some performance outcomes were impacted by possible learning effects over the study. Sleep disruption factors should be taken into account, especially in operational settings like the military where environmental factors (e.g., noise) disrupt sleeping conditions. Support Office of Naval Research, Code 34