The purpose of this study was to investigate the role of urea-N recycling, rumen bacterial community, and rumen epithelial gene expression in nitrogen utilization of calves. Here, 12 Holstein calves were divided into two dietary treatments: a high-protein diet (HP, 173 g/kg), and a low-protein diet (LP, 125 g/kg). Urea-N kinetics was evaluated using urea-15N15N isotope labeling method. Gene expression in rumen epithelium and liver, bacterial diversity, and metabolites in rumen were characterized using transcriptomic,...