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Background: Physiological mechanisms explaining why cardiorespiratory fitness (CRF) predicts cardiovascular morbidity and mortality are incompletely understood. We examined if CRF modifies vagally mediated heart rate variability (HRV) during acute physical or psychosocial stress or night-time sleep in adults with cardiovascular risk factors., Methods: Seventy-eight adults (age 56 years [IQR 50-60], 74% female, body mass index 28 kg/m 2 [IQR 25-31]) with frequent cardiovascular risk factors participated in this cross-sectional study. They went through physical (treadmill cardiopulmonary exercise test [CPET]) and psychosocial (Trier Social Stress Test for Groups [TSST-G]) stress tests and night-time sleep monitoring (polysomnography). Heart rate (HR) and vagally mediated HRV (root mean square of successive differences between normal R-R intervals [RMSSD]) were recorded during the experiments and analyzed by taking account of potential confounders., Results: CRF (peak O2 uptake) averaged 99% (range 78-126) in relation to reference data. From pre-rest to moderate intensities during CPET and throughout TSST-G, HR did not differ between participants with CRF below median (CRFlower) and CRF equal to or above median (CRFhigher), whereas CRFhigher had higher HRV than CRFlower, and CRF correlated positively with HRV in all participants. Meanwhile, CRF had no independent associations with HR or HRV levels during slow-wave sleep, the presence of metabolic syndrome was not associated with recorded HR or HRV levels, and single factors predicted HRV responsiveness independently only to limited extents., Conclusions: CRF is positively associated with prevailing vagally mediated HRV at everyday levels of physical and psychosocial stress in adults with cardiovascular risk factors.