Silver nanoparticles (AgNPs) synthesized by an implicitly environmentally gracious route using Acanthospermum hispidum aqueous leaves extract at room temperature reported. This approach is facile, swift, cost-effective and stable for a long time, reproducible at room temperature and in an eco-friendly manner to obtain a self-assembly of AgNPs. These fabricated AgNPs were investigated by ultraviolet-visible spectroscopy, field emission scanning electron microscopy, energy-dispersive X-ray spectroscopy, transmission electron microscope with selected area electron diffraction, X-ray diffraction, Fourier transform Infrared spectroscopy, particle size, zeta potential, photoluminescence with fluorescence lifetime spectroscopy and surface area with porosity studies of synthesized nanoparticles were analyzed by Brunauer-Emmett-Teller and Barrett-Joyner-Halenda curve. Besides, these AgNPs displayed antibacterial, antifungal antimycobacterial and antimalarial activity against some bacterial pathogens. From the outcomes obtained it is suggested that AgNPs could be used effectively in future nanobiotechnology and medical concerns.