Colletotrichum gloeosporioides, obtained from anthracnose symptoms on mango fruits. The disease infected mangoes were collected and isolated with confirming test. Bio-agents were applied in in vitro against C. gloeosporioides isolated from anthracnose affected mango. The study investigates to appropriate natural management. Three fungal bio-agents Aspergillus sp., Pestalotia sp. and Trichoderma sp. were used. Among the bio-agents, significantly highest mycelia growth inhibition 74.52% was achieved by Aspergillus sp. followed by 61.54% and 56.25% from Pestalotia sp. and Trichoderma sp. respectively over control. However, the present study suggested that Aspergillus sp. may use in controlling anthracnose of mango as bio-safety tools of integrated disease management.