The onset of pregnancy may be associated with hormonal changes and decline in milk yield of buffaloes. To investigate this, forty lactating buffaloes from 1st to 23rd weeks post-conception were selected. The animals were assigned to three treatments: PRT, PRS, NPRT and three milk yielding groups HMY, 66 to 75 l/week, n = 12; MMY, 56 to 65 l/week, n = 16; LMY, 46 to 55 l/week, n = 12). Milk samples were collected on alternate weeks and analyzed with ultrasonic milk analyzer. EIA was used for milk progesterone concentrations. Group means were compared and correlation analysis was conducted. Progesterone concentrations increased in almost similar pattern with the advancing weeks post-conception. The high and low yielder showed greater progesterone concentrations in the supplemented than the animals on traditional ration (P < 0.001). Progesterone concentrations correlated positively with fat (%), negatively with milk yield, protein (%) and lactose (%). Decline in milk yield became drastic when progesterone concentrations rose above 6.44 ng/ml. The pregnant animals on traditional ration exhibited a sharper decline in milk yield with the increasing progesterone concentrations as compared to pregnant animals with supplemented ration. It is concluded that concentrates supplementation induced a raise in progesterone levels. Progesterone concentrations and milk yield showed an inverse relationship.