Lactating Murrah buffaloes selected from the institute herd were categorized as hand and machine milking group. Blood samples were collected before milking, after concentrate feeding and teat message, during milking, and after completion of milking. Plasma prolactin (PRL), growth hormone (GH) and cortisol (CORT) concentrations were measured. Mean milk fat, protein, lactose and SNF content varied non-significantly between the groups. Milk somatic cell count (SCC), plasma glucose and NEFA concentration was more (P< 0.05) in machine milking group than in hand milking group. Both hand and machine milking operation significantly released plasma PRL (P< 0.01), CORT (P< 0.05) and GH (P< 0.01) hormone. Feeding of concentrate mixture resulted in a significant release in PRL and CORT hormone and their concentration increased further after teat message. Plasma PRL, CORT and GH varied significantly (P< 0.01) before, during and after milking (P< 0.01), and levels were normal 20-minutes post-milking in both the groups. It was concluded that feeding of concentrate mixture, teat message and milking operations either by hand or machine significantly stimulate release of plasma PRL, CORT and GH hormone in Murrah buffaloes.