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The present study examined the eusocial organization and mound architecture of fungus-growing Macrotermes termites and evaluated the biocontrol potential of fungal antagonists against both termites and their symbiotic fungus Termitomyces in Northern Sri Lanka. Field observations revealed large, dome-shaped mounds with efficient ventilation systems and well-organized internal chambers that maintained stable microclimatic conditions for fungal cultivation, while morphometric analysis of fungal combs indicated significant variability in size and weight, reflecting different developmental stages and efficient resource allocation by worker termites. Termitomyces was successfully isolated and identified based on characteristic basidiomycete features, and dual culture assays demonstrated strong antagonistic activity of Trichoderma viride, achieving 83.33% inhibition of radial growth after 14 days. An eco-friendly substrate formulation containing tamarind and fenugreek seeds supported maximum growth and sporulation of T. viride under optimal pH conditions, with greater colonization observed in sterilized termite combs compared to natural combs. Laboratory bioassays further revealed significant termite mortality caused by entomopathogenic fungi, particularly Paecilomyces cerevisiae, followed by Metarhizium anisopliae and Beauveria bassiana, highlighting the potential of fungal-based biocontrol agents as sustainable and environmentally friendly alternatives for Integrated Pest Management of termites.emphasized text
