Cyclotricium origanifolium is a plant belonging to the Lamiaceae family and is a species that grows in the Western and Southern Anatolian regions of Turkey. In our study, the antimicrobial activities of silver nanoparticles (AgNP) were investigated through Cyclotricium origanifolium plant extract. Characterization processes of the obtained AgNPs, suitable spectral analysis methods; Uv-Vis was determined by FT-IR, SEM-EDX, XRD. According to the results of the analysis, it was determined that the AgNPs were spherical in shape and had an average diameter of 17.60 nm. The antimicrobial effect of AgNPs was determined by the minimum inhibition concentration (MIC) method. Gram positive as test microorganisms; Staphylococcus aureus, Bacillus subtilis and gram negative; Escherichia coli, Pseudomonas aeruginosa bacteria, and Candida albicans fungal pathogen species were used. The suppression of microorganism growth was investigated by comparing the efficacy of standard antibiotics used in our study with AgNPs produced by the green synthesis method. It has been observed that the obtained AgNPs have a very strong effect on gram-positive B. subtilis and gram-negative E. coli bacteria, and are more effective against C. albicans than the normal antifungal drug. It was determined that the antimicrobial activity of AgNPs produced from C. origanifolium L. plants showed a stronger effect than standard antibiotics.