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The present study investigated changes in autonomic nervous system activity and emotions after a short (2 h) forest bathing program in the Xitou Nature Education Area (XNEA), Taiwan. One hundred and twenty-eight (60.0 ± 7.44 years) middle-aged and elderly participants were recruited. Physiological responses, pulse rate, systolic and diastolic blood pressure, heart rate variability (HRV), and psychological indices were measured before and after the program. We observed that pulse rate, systolic and diastolic blood pressure were significantly lower after the program, which indicated physiological benefits from stress recovery. The Profile of Mood States negative mood subscale scores of “tension-anxiety”, “anger-hostility”, “fatigue-inertia”, “depression-dejection”, and “confusion-bewilderment” were significantly lower, whereas the positive mood subscale score of “vigor-activity” was higher. Furthermore, participants exhibited significantly lower anxiety levels according to the State-Trait Anxiety Inventory. However, changes in sympathetic and parasympathetic nerve activity were nonsignificant. Our study determined that the short forest bathing program is a promising therapeutic method for enhancing heart rate and blood pressure functions as well as an effective psychological relaxation strategy for middle-aged and elderly individuals.