The present research examined the degree to which facets of trait mindfulness were associated with level and changes in psychological distress in response to a repeated carbon dioxide (CO2) breathing challenge. Undergraduate students (N = 93) completed a self-report measure of mindfulness and underwent two 7.5% CO2 challenges, spaced 1 week apart. Subjective distress, physical/fear symptoms, and threat cognitions were assessed at multiple times throughout each administration. A pattern emerged such that although mindfulness facets were not reliably associated with distress at either administration separately, a low (but not high) level of mindfulness was associated with a significant decrease in distress across administrations, likely indicative of habituation, for the facets Describing (β = − 0.25, p < − 0.01), Acting with Awareness (β = − 0.27, p < .01), and Observing (β = − 0.25, p < 0.01). This suggests that those components of mindfulness tied to noticing/attending to the present moment may at times interfere with typical habituation processes. In contrast, those high in components of mindfulness tied to not evaluating—Non-judging (β = − 0.23, p < 0.01) and Non-reacting (β = − 0.12, p < 0.01)—tended to report less distress, as expected. Findings suggest that the relation between trait mindfulness and stress response is more complex and nuanced than previously thought and that focusing on both mindfulness facets and repeated exposure to stressors may help elucidate this relationship.