This study explores the green synthesis of copper nanoparticles (Cu-NPs) using Panchagavya, a traditional organic substance composed of five cow-derived components, as a reducing and stabilizing agent. Cu-NPs were characterized using UV-Vis, FT-IR, XRD, SEM-EDS, TEM, DLS, and zeta potential analysis, which revealed their size, shape, and elemental properties. They exhibited strong antibacterial activity against Bacillus cereus and Pseudomonas aeruginosa. Biofilm inhibition was observed at various concentrations, with 38.98% at ½ MIC, 67.48% at MIC, and 84.03% at 2× MIC. SEM analysis confirmed that Cu-NPs disrupted the bacterial cell membrane, causing leakage of cellular contents. Antioxidant assays (DPPH, FRAP) revealed high scavenging activity, with percentages of 88.50% and 92.54%, respectively. Cu-NPs showed anticancer activity on MCF7 cells, with an IC