Fish mortality generally occurs during extreme summer temperatures in India which are apprehended to be more frequent in near future and may reduce the fish population, particularly in closed aquatic systems. This present study is conducted with the objectives to find out heat shock and associated oxidative stress responses that occurred in selected fish Labeo rohita due to extremely high water temperature (treated, 37-38 °C against control, 28-30 °C) exposure for 2 weeks. Calculated mortality was 30% during the experimental period. The results revealed the biomolecules associated with both the anti-oxidative response (reduced glutathione in serum, liver, muscle; catalase activity in liver, muscle; superoxide dismutase gene expression in the liver) and the heat shock response (hsp70 gene expression in the liver) were elevated under thermal stress. Pro-inflammatory responses (expression of complement protein 3, glyceraldehyde 3-phosphate dehydrogenase in the liver) and oxidative damages (lipid peroxidation in all studied tissue and DNA fragmentation in the liver) were more under thermal stress. Extreme thermal stress induced by partial lethal temperature exposure in this study led to the activation of both the heat shock response and the anti-oxidative response. However, these responses were not elicited to the level so that they can protect from oxidative damages and inflammation in the liver of all the studied fish that caused partial mortality in fish. Thermal stress-induced hepatotoxicity caused fish death which was documented for the first time in freshwater fish.