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Little millet (Panicum sumatrense Roth ex Roem. and Schult.) is one of the hardiest minor cereal crop indigenous to Indian sub-continent. The crop is cultivated by tribal and poor farmers for food and feed. Leaf blight disease has been a major production constraint and fungicidal sprays for the management of the disease may not be economically viable and feasible. Hence, the present in vitro study was carried out to know the antifungal activity of six fungal and 10 bacterial bio-agents against Alternaria alternata inciting little millet leaf blight disease. Among the fungal bio-agents, Trichoderma harzianum (ThB5) and among the bacterial bio-agents, Bacillus velezensis (P42) showed mycelial growth inhibition of 75.18 and 84.75%, respectively.