This study isolated and characterized a novel lytic bacteriophage, vB_EfS_C1, targeting Enterococcus faecium, and evaluated its antibacterial and antibiofilm activities against both vancomycin-susceptible and vancomycin-resistant strains. The phage exhibited strong lytic activity, broad stability across temperature and pH ranges, and its genome lacked virulence, lysogeny, and antibiotic resistance genes. Furthermore, it significantly inhibited bacterial growth and reduced biofilm biomass and viable cell counts, highlighting its potential as a safe and effective biocontrol and therapeutic agent against multidrug-resistant E. faecium.