The utilization of waste biomass, such as chicken manure (CM), for producing valuable products like fermentable sugars has gained increasing research attention. However, limited studies have explored the effect of acid pretreatment on sugar recovery efficiency specifically from CM. This study investigates the production of glucose and xylose from CM using dilute sulfuric acid (H₂SO₄) at concentrations of 0.6, 0.8, and 1.0 M under varying conditions. The results indicate that the highest yields were achieved from decrystallized CM, producing 46.21 mg glucose/g CM and 8.47 mg xylose/g CM under optimal conditions of 0.6 M H₂SO₄ and 100°C. In contrast, non-decrystallized CM yielded 13.98 mg glucose/g CM and 1.67 mg xylose/g CM under 1.0 M H₂SO₄ and 100°C. The decrystallization process using concentrated sulfuric acid effectively disrupted the lignin structure and partially hydrolyzed hemicellulose, enhancing cellulose accessibility during subsequent dilute acid hydrolysis. The study also revealed that glucose and xylose yields decreased as the dilute acid concentration increased from 0.6 to 0.8 M and temperature rose from 80 to 100°C for decrystallized CM. Conversely, for non-decrystallized CM, sugar yields increased with higher acid concentration and temperature. These findings highlight the critical role of pretreatment in improving sugar recovery from CM and suggest that optimizing acid concentration and thermal conditions can enhance the efficiency of biomass conversion. This research contributes to the sustainable valorization of agricultural waste into bio-based products.