Background: Manual breathing assist technique (MBAT) is a common physical therapy technique used to facilitate airway clearance and improve ventilation and oxygenation. The effects during and immediately after intervention in individuals with chronic obstructive pulmonary disease (COPD) are unknown. This study aimed to investigate the acute effects and potential mechanisms of MBAT on lung volume, dyspnea, and oxygenation in individuals with COPD. Methods: This non-randomized quasi-experimental pre-test/post-test study included participants from pulmonary rehabilitation programs at Tagami Hospital (COPD group) and a community exercise program (Healthy group). During a single session, MBAT was applied during the expiration of every breath for 10 minutes. Dyspnea and lung volumes (tidal volume; VT, inspiratory capacity; IC, inspiratory reserved capacity; IRV, expiratory reserve capacity; ERV) were collected at baseline and after MBAT. Pulse oximetry (SpO2), skeletal muscle oxygenation (SmO2), and oxy- and deoxy-hemoglobin (O2Hb and HHb) using near-infrared spectroscopy (NIRS) were collected at baseline, during, and after MBAT. Between-group comparisons were conducted using the Mann-Whitney U-test and chi-square analyses. Within-group changes before and after MBAT were analyzed using the Wilcoxon signed-rank test. The Kruskal-Wallis test was used to detect differences in NIRS variables in each phase and over time. Results: Thirty participants with COPD, matched for age and sex, were included, with 15 individuals per group. The difference scores of VT, IRV, and IC were significantly higher in the Healthy group than in the COPD group, but improvements in dyspnea and SpO2 were significantly higher in the COPD group. Compared to baseline, ERV decreased significantly in both groups, with dyspnea and SpO2 improving significantly only in the COPD group. Inspiratory accessory muscle ΔO2Hb and ΔHHb were significantly higher and lower (respectively) during MBAT in the COPD group compared to the Healthy group. Additionally, only the COPD group had increased SmO2 during and after MBAT compared to baseline. Conclusions: MBAT in patients with COPD had acute physiological effects in reducing dyspnea by facilitating expiration and decreasing the recruitment of accessory respiratory muscles. MBAT may help individuals with COPD reduce dyspnea before exercise therapy in a pulmonary rehabilitation program.