Gentamicin (GM) is an aminoglycoside antibiotic commonly used against life threatening gram negative bacterial infections, however, nephrotoxicity remains the major concern for its long term use. Although its effects on kidney are well characterized but there have been no studies regarding its effects on intestine. We hypothesize that GM causes adaptive coordinated effect on enzymes of carbohydrate metabolism and terminal digestion/ absorption in rat intestine. Rats were administered a nephrotoxic dose of GM (80 mg /kg body weight) daily for 15 days and a time dependent effect was observed on various enzyme activities. Activities of lactate (LDH), malate (MDH) and isocitrate (ICDH) dehydrogenases, significantly increased and peaked at different time intervals of GM treatment. Whereas LDH activity remained higher, MDH and ICDH activity slowly declined from their peak values. Activities of fructose-1,6-bisphosphatase, glucose-6-phosphatase and glucose-6-phosphate dehydrogenase increased but malic enzyme decreased in a time dependent manner. Activity of alkaline phosphatase and sucrase significantly increased but gamma-glutamyl transpeptidase activity decreased. GM administration increased lipid peroxidation, glutathione peroxidase but decreased superoxide dismutase and catalase activities. The results indicate that GM treatment selectively upregulated certain enzymes of carbohydrate metabolism and terminal digestion/absorption and perturbed antioxidant defenses.