Abstract
Shift workers are at elevated risk of chronic diseases due to circadian rhythm disruption, suboptimal lifestyle behaviors, and potentially altered gut microbiota (GM). This study investigated variations in GM diversity and composition across three weekly shifts in rotating shift workers and following walnut supplementation. Using a within-person design, GM diversity and composition were compared in 13 shift workers during morning, afternoon, and night shifts. After a three-week observational period, participants added a daily serving of walnuts to their habitual diet for an additional three weeks. GM was analyzed via 16S rRNA sequencing, assessing diversity and bacterial composition across shift types and between the observational and interventional phases. Overall GM composition did not differ between the beginning and end of shifts, by shift type, or following walnut supplementation. Bacterial diversity remained stable except for a significant decrease at the end of the night shift during observation (p = 0.03), which was not observed during walnut supplementation. GM clustered strongly by subject, and a healthier diet correlated with greater mean GM diversity (r = 0.64, p = 0.02). Despite overall GM stability, the decline in diversity during night shifts suggests that targeted nutritional strategies, such as walnut supplementation, may help preserve gut health in shift workers.