The objective of the study was to investigate the effects of inclusion of Pleurotus florida treated wheat straw in the total mixed rations (TMRs) on feed intake, growth performance, nutrient digestibility, and nitrogen retention in male buffalo calves. As a pilot study, four TMRs, i.e., TMR1 having 0% P. florida treated wheat straw (FTWS), TMR2 (20% FTWS), TMR3 (40% FTWS), and TMR4 (60% FTWS) with berseem hay as basal diet, were formulated. Sixteen Nili-Ravi male buffalo calves (aged 10-12 months, weighing 73 ± 2.50 kg) were divided into four equal groups and randomly assigned one of four TMRs. A significant increase (P < 0.05) was observed in all nutrients intake, their digestibility, weight gain, and nitrogen retention with TMRs incorporated with FTWS. Highest feed conversion ratio (FCR) of 2.63 was noted with TMR1-0% and the lowest FCR (1.80) with TMR4-60%, on the other hand. In conclusion, the TMR4 (60% FTWS) has the potential to increase the weight gain, nutrient digestibility, nitrogen retention, and feed efficiency in buffalo calves. Therefore, inclusion of 60% Pleurotus florida treated wheat straw is recommended as TMRs with berseem hay based basal diet for feeding buffaloes calves.