Background Green synthesis of silver nanoparticles (AgNPs) using plant extracts has gained increasing attention as an environmentally friendly alternative to conventional chemical methods. Salix alba L contains bioactive phytochemicals that can act as reducing and stabilizing agents during nanoparticle synthesis. This study aimed to synthesize AgNPs using an aqueous bark extract of S. alba and to evaluate their antimicrobial and cytogenetic effects. Methods Silver nanoparticles were synthesized by mixing S. alba aqueous extract with silver nitrate solution. Nanoparticle formation and surface characteristics were assessed using ultraviolet visible spectroscopy, atomic force microscopy (AFM), and scanning electron microscopy (SEM). Antimicrobial activity against Escherichia coli, Staphylococcus aureus, and Candida albicans were evaluated using the agar well diffusion method. Cytogenetic effects were assessed in mice bone marrow cells by measuring the mitotic index. Results AFM and SEM analyses showed that AgNP size and morphology varied with silver nitrate concentration, with smaller and more uniform nanoparticles observed at lower concentrations. The synthesized AgNPs exhibited antimicrobial activity against all tested microorganisms, with inhibition zones increasing in a dose dependent manner. Cytogenetic analysis demonstrated a significant reduction in mitotic index in AgNP treated groups at higher concentrations, while S. alba extract alone did not induce genotoxic effects and maintained normal or slightly elevated mitotic index values. Conclusions Salix alba L mediated silver nanoparticles demonstrated antimicrobial activity and concentration dependent cytogenetic effects. These findings indicate the importance of careful dose optimization when considering biologically synthesized AgNPs for potential applications.