The eco-friendly and cost-effective strategies for the preparation of nanoparticles are the cutting edge researches in the field of nanotechnology. The objective of the study was the eco-friendly synthesis, characterization and their antibacterial activity of AgNPs using stem extract of Phyllanthus pinnatus. The appearance of the dark brown color showed the biofabrication of AgNPs and its characterised by UV-Vis, SEM, XRD, and FT-IR analysis. Consequently, the antibacterial activity of AgNPs was tested against different bacterial pathogens. The UV-Vis spectrum of colloidal AgNPs exhibited a strong absorption peak at 490 nm. Besides, the SEM image showed that the AgNPs were fairly uniform in the nanoscale with mostly cubical morphology. The XRD analysis showed the crystalline nature of AgNPs. In addition, the FT-IR spectrum revealed the presence of functional groups attached to the surface of AgNPs that may have a role in bioreduction and stabilization of AgNPs. The AgNPs showed dose-dependent toxicity against bacterial pathogens. The maximum zone of inhibition was found at 1.8 mm in diameter for both Vibrio cholera and Shigella flexneri at 40 mu L concentration. Although the current study showed potent in vitro antibacterial activity of AgNPs, further studies are required to establish the bactericidal potential of AgNPs on animal models for drug development.