The main objective of this this cross-sectional study was to analyze the influence of lifestyle fac-tors (diet, physical activity, sleep), which can affect the concentrations of fecal short-chain fatty acids (SCFAs) and SCFAs potential role in modulating cardiometabolic disease risk by interact-ing with biochemical and body composition parameters. The study comprised 77 healthy, non-obese individuals, aged 30-45 years who were assessed in the light of SCFAs concentration in stool, diet, physical activity level and sleep duration. Moreover, body composition measurement and patients’ biochemical parameters were included in the analysis. We have indicated a signifi-cant negative correlation between several SCFAs [especially acetic acid (AA), isobutyric acid (IBA), butyric acid (BA), isovaleric acid (IVA) and valeric acid (VA)] with BMI, VAT/SAT ratio (visceral to subcutaneous fat ratio) and percentage of fat mass in a group of females enrolled in the study as well as with waist circumference (WC) in case of both sexes included in the study. Moreover, the results of our study acknowledged the importance of a diet in shaping SCFAs pro-file - we noticed the significant negative associations between energy and fat intake and some SCFAs in males [IBA, IVA, VA, isocaproic acid (ICA)]. In terms of fiber intake, we noticed elevat-ed concentrations of vast majority of SCFAs and amount of SCFAs in total in case of both sexes included in the study. These strong correlations reflect the fact that diet shapes the composition of the gut microbiota and SCFAs (main microbial metabolites) are synthesized from dietary fiber. Furthermore, we noticed that in a group of women, the concentration of AA, propionic acid (PA) and ICA as well as the total concentration of SCFAs showed a significant positive association with their sleep duration. We concluded that SCFAs can have a potential role in modulating car-diometabolic disease risk by interacting with adiposity parameters and diet. In addition, this potential direct link between diet and SCFAs may at least partly contribute to sleep improvement.