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This cross-sectional study examined the relationship between daily physical activity, short-chain fatty acids (SCFAs) concentration, and abundance of SCFA-producing bacteria in 30 healthy young adults (15 males and 15 females) aged 20-23 years. Participants provided fecal samples for gut microbiota and SCFA analysis. Daily physical activity, including step count and brisk walking time at an intensity of ≥ 4 metabolic equivalents (METs), was assessed. Participants were categorized into two groups (more and less active) based on their daily brisk walking time at a 4METs intensity and step count. We compared SCFA concentration and abundance of SCFA-producing bacteria between the groups and explored the relationship between the abundance of SCFA-producing bacteria and daily physical activity levels. No significant differences in the major SCFAs (acetic acid and butyric acid) were observed between the groups. A lower abundance of Ruminococcaceae, Faecalibacterium, and Faecalibacterium prausnitzii were observed in the active group. Overall, the relative abundance of intestinal Eubacterium, contributing to butyrate concentration, was associated with daily brisk walking at a 5METs intensity. In conclusion, while daily brisk walking did not directly affect SCFA concentration, it may be associated with the relative abundance of intestinal Eubacterium in healthy young adults. [ABSTRACT FROM AUTHOR]