The effect of different ratios of urea to amino acid N at a fixed concentration of soluble sugars as energy source and varying levels of soluble sugars at optimum urea cell suspension was obtained from the rumen fluid of buffalo (Bubalus bubalis) calves fed on a growth ration. Under glucose fermentation, the bacterial protein content of the incubation mixture (I. M.) was increased to 3.91, 6.31 and 5.08 times the control value (urea alone) when 25, 50 and 75% of urea-N was replaced with amino acid N, respectively. With cellobiose, the corresponding increase was 4.06, 5.29 and 5.63 times. At 50% urea-N replacement with amino acid N (a ratio for maximum protein synthesis), the bacterial content was maximum when 1 g glucose or cellobiose per 100 ml of I. M. was added. Per cent incorporation of radioactivity from amino acids into bacterial protein was maximum at 25% amino acid N level with both the soluble sugar sources. The total amino acids incorporated into bacterial protein were, however, more at 50% than at 25% amino acid N level.