Different agroclimatic conditions affect the endocrine system and metabolism of animals. This study aimed to analyze the effect of different agroclimatic conditions on blood thyroxine, cortisol, and testosterone concentrations in Swamp buffaloes. For this study, hormonal profiles of 38 Swamp buffaloes, consisting of 18 buffalo bulls aged 3-5 years and 20 heifers aged 2-3 years, kept under different agroclimatic conditions were analyzed. These animals were sampled from three districts in West Sumatra, namely: Lubuk Basung district (representing lowland conditions), Akabiluru district (representing moderate-land conditions) and Matur district (representing highland conditions). Hormone analysis was performed at the integrated research laboratory of the Syahkuala University in Banda Aceh, using the ELISA method. The parameters measured were the ambient temperature, temperature-humidity index (THI) and concentrations of thyroxine, cortisol and testosterone hormones. The results showed non-significant differences in serum thyroxine, cortisol and testosterone concentrations in Swamp buffaloes of lowland, moderate land and highland. In conclusion, differences in agroclimatic conditions did not noticeably influence the endocrine system and metabolism of the Swamp buffaloes.