Green synthesis is one of the fastest and best ways for ecofriendly nanoparticle synthesis. This study aims to investigate the use of the green microalgaeParachlorella kesseleriandCyclotellaspp. for the biological synthesis of silver nanoparticles (AgNPs). This work focuses on optimizing various parameters necessary for the production and stability of AgNPs. The nanoparticle formation was confirmed by UV-Visible analysis, which revealed the surface plasmon resonance band at 420 nm. The characterization of the AgNPs was performed using UV-visible spectroscopy, X-ray diffraction (XRD), scanning electron microscopy coupled with energy dispersive X-ray microanalysis (SEM-EDS), Fourier transform infrared spectroscopy (FTIR), and inductively coupled plasma atomic emission spectroscopy (ICP-AES). The antimicrobial properties of these bioactive AgNPs were also tested, showing excellent antibacterial activity against six bacterial strains,Escherichia coli, multidrug-resistantEscherichia coli,Bacillus clausii,Pseudomonas aeruginosa,Staphylococcus aureus, andSalmonella typhi. The biosynthesis of AgNPs from living cultures of microalgae has remarkable antibacterial properties. Other studies are underway in our laboratory to clarify the mechanism of the biosynthesis of these nanoparticles, and their action on bacteria.