Longitudinal analysis of the hand microbiome in response to chlorine-based antiseptic use during a military field exercise

Hand hygiene is essential for infection control, yet the impact of frequent antiseptic use on the skin microbiome, which is crucial for skin barrier function and pathogen exclusion, remains underexplored, especially in field conditions. In high-risk military settings, there is a need for safe, multipurpose antiseptics that avoid the drawbacks of alcohol-based options, including skin irritation, flammability, and unsuitability for wound application. We conducted a longitudinal study during the Norwegian-led military exercise Cold Response 2022 to assess the effects of repeated use of stabilized hypochlorous acid, a non-alcoholic antiseptic, on the hand microbiome of soldiers in a field environment. Participants used either the chlorine-based antiseptic (n = 20) or standard hygiene practices, mainly alcohol-based sanitizers (n = 19), for 10 days. Skin swabs from hands and untreated forearms were collected at baseline, post-intervention, and three weeks after discontinuation, and analyzed using 16S rRNA sequencing. Overall, our data revealed that field exposure accounted for more variation in bacterial composition than antiseptic use. When comparing alpha and beta diversity trajectories between the two antiseptic groups, there was minimal divergence during active field use, but a significant post-treatment increase in alpha diversity associated with the use of chlorine-based antiseptic. In contrast, untreated forearms showed no such group-level differences. These findings suggest that antiseptic-driven microbiome shifts are most apparent during recovery, with non-alcoholic formulations supporting faster recolonization after treatment. Stabilized hypochlorous acid represents a promising alternative to standard alcohol-based antiseptics for military personnel. IMPORTANCEEffective hand hygiene reduces pathogen transmission, but its influence on the skin microbiome has rarely been studied. Our research emphasizes the importance of longitudinal assessments by identifying greater antiseptic-driven differences occurring after cessation of treatment than during repeated use. Service members frequently use antiseptics in challenging environments, highlighting the operational need for a non-alcoholic, multipurpose product that can be safely used on both intact skin and wounds. Our findings suggest that a chlorine-based non-alcoholic antiseptic permits faster recovery of bacterial diversity after perturbation, informing product selection and hygiene policies in defense and other high-risk settings.