Background: Nowadays, the environmentally friendly procedures must be developed to avoid using harmful compounds in synthesis methods. Their increase interest in creating and researching silver nanoparticles (AgNPs) because of their numerous applications in many fields especially medical fields such as burn, wound healing, dental and bone implants, antibacterial, viral, fungal, and arthropodal activities.  Biosynthesis of nanoparticles mediated pigments have been widely used as antimicrobial agent against microorganisms. Silver nanoparticles had synthesized by using melanin from locally isolate Pseudomonas aeruginosa, and used as antimicrobial activity against pathogenic microorganisms. Aim of the study: Isolation of Pseudomonas aeruginosa that produce melanin and extraction of melanin. Synthesis and characterization silver nanoparticle and study of the antimicrobial activity of silver nanoparticles in the presence of melanin against UTI pathogens. Materials and methods: The samples swab inoculated on cetrimide agar as selective media and incubated aerobically for 24 hours at 37 °C. Used nutrient agar with nutrient broth supplement with 1% tyrosine for screening for melanin production by P. aeruginosa isolates,silver nanoparticles synthesis from P. aeruginosa was done according to biological method and was characterized with AFM, UV-Visible, XRD, FTIR and FE-SEM. Agar well diffusion method was used to examine the effect of combination against UTI pathogens. Results: The synergistic effects of AgNPs and melanin were evaluated to compare between the two treatments (silver nanoparticles alone and combination of silver nanoparticles and melanin). The results revealed that the combination showed the highest antimicrobial activity in compare with silver nanoparticles alone.