Nowadays nanoparticles are used in many fields of life all over the world, and there are numerous ways to obtain them: chemical, physical and biological processes. In recent times, the biological method for the synthesis of nanoparticles associated with using plant extract is widely spread. Optimal conditions for synthesis of silver nanoparticles using aqueous seeds extract of Myristica fragrance were highlighted in this research, such as type of plant extract, weight of extracted plant material, volume ratio of plant extract to AgNO3 and temperature of reaction. The study proved that the optimal status for AgNPs synthesis by using 10 g of M. fragrance seeds powder were added to 100 mL boiled distilled water, then homogenized and filtered after 24 hours. Aliquot of 5 mL of hot aqueous extract were added to 45 mL of 1*10-3 M AgNO3 solution in the water bath with a magnetic stirrer for the bio-reduction process at 60 °C. The biological activity of AgNPs nanoparticles was evaluated by using well diffusion method and biofilm formation for G+ and G- bacteria including Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis and Klebsiella pneumonia, while the effect of AgNPs nanoparticles on plasmid curing was investigated for Escherichia coli and Staphylococcus aureus only. Atomic Force Microscopy (AFM) images proved that Ag particles are in nanometer- size and have granular shape, the size of silver nanoparticle is (74.55 nm) for the sample taken after 16 min of the reaction. Nanoparticles of various concentrations have proven effective in inhibiting bacterial growth after antimicrobial activity test, biofilm formation and plasmid curing as they exhibited a remarkable effect in inhibiting the growth of both Gram-positive and negative bacteria