Domestic wastewater contains numerous pathogens that lead to waterborne diseases. Therefore, ensuring effective disinfection during wastewater treatment is essential for protecting public health. However, conventional disinfection approaches such as chlorination, UV, and ozone have many drawbacks. Silver nanoparticles (Ag NPs) is promising alternative disinfectant for wastewater treatment. The use of biosynthetic Ag NPs is eco-friendly and cost-effective. This study investigates the synthesis of Ag NPs using the cell-free extract of three actinomycete strains (βB1, βC1 and UA15). The confirmation of Ag NPs formation was conducted through UV-visible spectroscopy, X-ray diffraction (XRD), and transmission electron microscopy (TEM). Domestic wastewater samples from various sewage treatment plants were treated with minimum inhibitory concentration (MIC) of Ag NPs and then analyzed for various physicochemical variables and microbial groups. Compared to control the treated samples characteristics complied with standards for discharging liquid wastes with reduction in microbial groups exceeded 99%. Different Ag NPs concentrations and exposure times were investigated to minimize time and concentration of Ag NPs while maintaining disinfection activity. Treatment with 15 µg/ml concentration of Ag NPs by βB1 was found efficient in the disinfection. The efficiency of the designated concentration versus UV and chlorination was compared. Although chlorination and UV eliminate all the targeted groups, a comparable disinfection capability of the tested Ag NPs was proved. In conclusion, the investigated Ag NPs hold significant promise as a cost-effective and eco-friendly disinfection solution, particularly in domestic wastewater treatment in compare to the common disinfection techniques such as chlorination or UV with less drawbacks.