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• Heart rate variability (HRV) can influence spatial navigation performance. • HRV levels particularly influence early stages of spatial processing. • High HRV individuals have fewer errors for locating stimuli on an 3D environment. • These effects are present even after controlling physical activity indicators. • A better cardiac health can result in more optimal spatial orientation skills. Heart rate variability (HRV) is considered one of the most relevant indicators of physical well-being and relevant biomarker for preventing cardiovascular risks. More recently, a growing amount of research has tracked an association between HRV and cognitive functions (i.e., attention). Research is still scarce on spatial orientation, a basic capability in our daily lives. It is also an important indicator of memory performance, and its malfunctioning working as an early sign of dementia. In this study, a total of 43 female students (M Age = 18.76; SD = 2.02) were measured in their lnRMSSD using the photoplethysmography technique with the Welltory smartphone app. They were also tested in their spatial memory with The Boxes Room, a virtual navigation test. Measures of physical activity were obtained with the International Physical Activity Questionnaire (IPAQ). Correlation analyses and repeated measures ANOVA were performed, comparing participants with high / low lnRMSSD in their spatial performance. Results showed that, at an equal level of physical activity, participants with a higher lnRMSSD were more effective in the early trials of The Boxes Room, being more precise in estimating the correct position of the stimuli. Moreover, a subsequent simple linear regression showed that a higher lnRMSSD was related to a smaller number of errors at the beginning of the spatial task. Overly, these results outline the relationship between HRV and navigation performance in early stages of processing, where the environment is still unknown and the situation is more demanding. [ABSTRACT FROM AUTHOR]