Background As a consequence of their eco-friendliness, simplicity and non-toxicity, the fabrication of metal and metal oxide nanoparticles using greener chemistry has been a highly attractive research area over the last decade. Aim In this study focused on the fabrication of silver-Zinc oxide nanocomposite (Ag-ZnO NCs) using Ruta chalepensis leaf extract and evaluating its potential biological activities, against Echinococcus granulosus in an in vitro and in vivo model using BALB/c mice. Methods In this study, the synthesis of Ag-ZnO NCs was accomplished using local R. chalepensis leaf extracts. The synthesized nanocomposites were identified using UV-Vis, SEM-EDX, XRD, and FTIR. For a short-term assessment of acute toxicity, BALB/c mice were given the prepared NCs orally. Dual sets of mice were also intraperitoneally injected with protoscoleces for secondary echinococcosis infection. Furthermore, a blood compatibility test was carried out on the nanocomposites. Results The synthesized Ag-ZnO NCs presented a surface plasmon peak at 329 and 422 nm. The XRD, SEM, and EDX confirmed the purity of the Ag-ZnO NCs. The FTIR spectra indicated the formation of Ag-ZnO NCs. Compared to the untreated infected mice, the treated-infected animals displayed an alteration in the appearance of the hepatic hydatid cysts from hyaline to whitish cloudy with a rough surface appearance. Lysis of RBCs at various doses of Ag-ZnONCs was significantly less than the positive contro,. Conclusion These findings revealed that the Ag-ZnO NCs didn’t cause any adverse symptoms and no mortality was observed in all administered groups of mice. The obtained outcomes confirmed that concentrations of up to 40 μg/mL of the bio-fabricated Ag-ZnONCs induced no notable harm to the red blood cells.