Agricultural soils subjected to repeated pesticide application accumulate chlorpyrifos residues that persist for months to years, posing significant risks to soil health, non-target organisms, and groundwater quality. Bioremediation using indigenous fungal isolates offers a practical and ecologically sound approach to address this contamination. This study isolated and identified chlorpyrifos-degrading fungi from pesticide-contaminated agricultural soil in Okra village, Balrampur district, Chhattisgarh, India. Serial dilution plating on Potato Dextrose Agar yielded a fungal count of 4.2 × 10⁴ CFU/g, from which three active degraders Aspergillus niger, Trichoderma viride, and Aspergillus terreus were confirmed by ITS region sequencing and NCBI BLAST analysis. Biodegradation was optimized across temperature, pH, nutrient concentration, aeration, and salt concentration. HPLC analysis over 30 days showed that T. viride achieved the highest degradation at 89.8%, followed by A. niger at 87.9% and A. terreus at 84.4%, compared to only 8.6% in the uninoculated control. First-order kinetic modeling confirmed half-lives of 8.5, 9.3, and 11.7 days respectively, representing up to a 34-fold reduction compared to abiotic degradation alone. These findings establish indigenous fungal isolates from Chhattisgarh as strong candidates for developing targeted bioremediation strategies against chlorpyrifos contamination in agricultural ecosystems.