The three-dimensional movement of the horse in physical therapy is a valuable kinesio-therapeutic phenomenon that simultaneously affects several body systems, including particularly the neuromuscular system. However, the effects of equine-assisted services (EAS) on neuromuscular activation patterns in older adults have not been thoroughly investigated. In this study, we evaluated the impact of a 10-weeks EAS program on trunk muscles in older adults who used a saddle and placed their feet in stirrups for the first 15 minutes and out of stirrups for the remaining 15 minutes of 30-minute EAS sessions. We gathered electromyographic (EMG) data of the trunk muscles five times each on the first, fifth, and 10th sessions: pre-EAS and post-EAS on a stationary horse and at 1-minute, 15-minutes, and 30-minutes on a horse in motion. Participants were 20 adults, aged 60–79 years. We analyzed normalized EMG data with 5 (session time) by 3 (session number) analyses of variance (ANOVAs) with repeated measures and with Bonferroni’s testing (p ≤ .05). There was a significant difference over the number of interventions for the right thoracic paravertebral muscle (p = .025) and session time effect for the left trapezius (p = .042), right thoracic paravertebral (p < .001), right and left multifidus (p < .001), and right and left rectus abdominis muscles (p < .001). Thus, trunk muscles in older adults showed complex neuromuscular activation synchronized with the horse’s movement, which was influenced by session time and number of interventions. The practical implication of these findings is that EAS can reduce fall risk among elderly adults of both sexes.