Along with the progress of nanoscience and nanotechnology came the means to synthesize nanometric scale materials. While changing their physical and chemical properties, they implicitly changed their application area. The aim of this paper was the synthesis of colloidal silver nanoparticles (Ag-NPs by ultrasonic disruption), using soluble starch as a reducing agent and further as a stabilizing agent for produced Ag-NPs. In this context, an important parameter for Ag-NPs preparation is the pH, which can determine the particle size and stability. The physical-chemical behavior of the synthesized Ag-NPs (shape, size, dispersion, electric charge) is strongly influenced by the pH value (experiment being conducted for pH values in the range between 8 and 13). The presence of a peak located at 412 nm into the UV-VIS spectra demonstrates the presence of silver nano-spheres into the produced material. In UV/VIS spectra, we observed a specific peak for yellow silver nano-spheres located at 412 nm. Samples characterization was performed by scanning electron microscopy, SEM, energy-dispersive X-ray spectroscopy, EDX, Fourier-transform infrared spectroscopy, and FT-IR. For all Ag-NP samples, we determined the zeta and observed that the Ag-NP particles obtained at higher pH and have better stability. Due to the intrinsic therapeutic properties and broad antimicrobial spectrum, silver nanoparticles have opened new horizons and new approaches for the control of different types of infections and wound healing abilities. In this context, the present study also aims to confirm the antimicrobial effect of prepared Ag-NPs against several bacterial strains (indicator and clinically isolated strains). In this way, it was confirmed that the antimicrobial activity of synthesized Ag-NPs was good against Staphylococcus aureus (ATCC 25923 and S. aureus MSSA) and Escherichia coli (ATTC 25922 and clinically isolated strain). Based on this observation, we conclude that the prepared Ag-NPs can represent an alternative or auxiliary material used for controlling important nosocomial pathogens. The fungal reference strain Candida albicans was more sensitive at Ag-NPs actions (zone of inhibition = 20 mm) compared with the clinically isolated strain (zone of inhibition = 10 mm), which emphasizes the greater resistance of fungal strains at antimicrobial agent’s action.