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River pollution is a major environmental concern affecting freshwater ecosystems worldwide. Biomolecular stress responses are increasingly recognized as sensitive indicators of sub-lethal pollution stress in aquatic organisms. The present study evaluates pollution-induced biomolecular stress responses in aquatic fauna of the Narmada River at Jabalpur, integrating physicochemical parameters with molecular biomarkers. Alterations in antioxidant enzyme activity, lipid peroxidation, and membrane stability were recorded at pollution-impacted sites, indicating oxidative and metabolic stress. These responses were strongly correlated with changes in dissolved oxygen and turbidity, demonstrating the biophysical linkage between environmental stressors and molecular damage. The study highlights the utility of biomolecular biomarkers as early warning tools for river health assessment.