AbstractAn experiment was performed to access the comfort level with the help of blood biochemical profile in buffalo’s heifers. Randomly eighteen healthy Murrah buffaloes of almost uniform in age, body weight and health status selected for experiment at Livestock Farm Complex (LFC), College of Veterinary Sciences & Animal Husbandry, DUVASU, Mathura up to three month duration. The selected animals were randomly divided into three groups viz. Concrete flooring (T1), Cow dung bed flooring (T2) and Rubber mat installed flooring. The study revealed that mean of total protein (mg/dl) in buffalo heifers were observed significantly (P>0.05) higher in T2 & T3 group (6.08 and 6.11) than T1 (5.45), while the total globulin (mg/dl) was significantly higher in T3 (2.60) than T1 & T2 (1.24 and 1.74) after 30 days of experimental period. The blood urea nitrogen (mg/dl), plasma creatinine (mg/dl), plasma glucose (mg/dl), plasma cholesterol (mg/dl) and plasma albumen (mg/dl) have on any significant changes during different phases of experimentation. Keywords: Heifer; Plasma; Lameness.