The persistent decline in biodiversity due to human activities is a major concern for ecosystems and society. The Atewa Forest Reserve (ATFR) in Ghana is a critical biodiversity hotspot, housing numerous butterfly species, including endemics and rare types, facing threats from human activities. This study assessed the different stress levels of anthropogenic activities on the population of butterflies in the ATFR. The three study areas considered were: Primary Forest, Secondary Forest, and Depleted Agricultural zones. Methods like transect counts, sweep nets, and bait traps were employed for species assessment. ANOVA analysis revealed significant population differences (p= 0.003, α = 0.05) between the areas. The Shannon-Wiener index indicates the highest diversity in Primary Forests (2.196), moderate in Secondary Forests (2.230), and least in Depleted Agricultural areas (2.273). Despite degradation in Secondary and Depleted Agricultural areas due to human influence, the ATFR maintained rich butterfly diversity. The study highlights the richness of butterfly diversity in ATFR, emphasizing the need for conservation efforts to mitigate anthropogenic effects. Recommendations include reclamation of encroached farming zones, sustainable practices, legal frameworks for biodiversity conservation, halting damaging activities within the reserve, and offering alternative livelihoods for local communities dependent on the forest.