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The purpose of this study was to investigate the effects of melatonin implants on biochemical metabolite composition and enzymatic properties of blood plasma of buffalo bulls in the non-breeding season. Ten Murrah buffalo bulls, divided into control and treated groups, each of five animals, were used during non-breeding season (July to September). Treated bulls were implanted with melatonin (18 mg of melatonin per 50 kg of body weight) for a period of two months. During non-breeding season, melatonin treatment resulted in increases (P<0.05) in total protein and creatinine of blood plasma, and decreases (P<0.05) in blood plasma high density lipoprotein-cholesterol (HDL), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities. In conclusion, melatonin treatment affected several metabolites in blood plasma of buffalo bulls during non-breeding season under tropical condition.