The present research deals with the development of a novel bioinspiredin situfabrication of reduced graphene oxide (rGO)-silver nanoparticle (AgNPs) nanocomposite (rGO@AgNCs) using microbes namelyPseudomonas aeruginosa(PA) andStaphylococcus aureus(SA). The fabricated rGO@AgNCs were characterized using Ultraviolet-visible (UV-Vis) spectroscopy, Fourier-transform infrared spectroscopy (FTIR), particle size analysis, polydispersity index (PDI), zeta potential analysis, energy dispersive x-ray analysis (EDAX), Raman spectroscopy, powder x-ray diffraction (PXRD), high-resolution transmission electron microscopy (HR-TEM) analysis, etc. Furthermore, the rGO@AgNCs-PA and rGO@AgNCs-SA interaction with serum protein, pH stability study, andin vitrodissolution of AgNPs were also performed. The research findings of the proposed study demonstrated the simultaneous reduction of graphene oxide (GO) and AgNPs and the formation of rGO@AgNCs in the presence of microbes. Thein vitrodissolution studies of rGO@AgNCs composites showed better AgNPs dissolution with controlled release and offered remarkable matrix integrity throughout the dissolution period. The size and stability of rGO@AgNCs-PA and rGO@AgNCs-SA had no significant changes at physiological pH 7.4. A minimal decrease in the zeta potential of rGO@AgNCs was observed, which may be due to the weak interaction of nanocomposites and albumin. The antibacterial application of the synthesized nanocomposite was evaluated against a pathogenic mastitis-forming bacterium. The obtained results suggested an admirable antibacterial activity of synthesized nanocomposites against the tested microbes. This knowledge will assist the scientific fraternity in designing novel antibacterial agents with enhanced antibacterial activity against various veterinary pathogens in near future.