The objectives of this study were to evaluate the contribution of urea on the nutritional quality and microbial community of ensiled alfalfa (Medicago sativa L.). Alfalfa silage was control group without urea (AL), supplementation with 0.5% urea (AU1), or supplementation with 1% urea (AU2). The silage tanks were opened and sampled after silage at 0, 15, 30, and 60 d. Results showed that AU2 had ... [Show full abstract] higher pH, ratio of (ammonia–N)/(total nitrogen) (NH3–N/TN) and crude protein (CP) content than those in AL and AU1, while AU1 had higher acid detergent fiber (ADF) than that in AL and AU2 after 15 d silage. Richness and diversity indices of microbial communities in silage were no significant differences among AL, AU1 and AU2 group. Proteobacteria (58.23%) and Firmicutes (40.95%) were the predominant phylum in three groups during the silage process. The percent of community abundances on genera level of Enterobacteriaceae (37.61%) and Klebsiella (41.78%) in AL were a little higher than those in AU1 (30.39%, 25.02%) and AU2 (33.48%, 26.92%). These results showed that silage with urea alone could not improve the quality of alfalfa.