Although toxoplasmosis is worldwide spread, the available medications have low efficacy and high toxicity. This research aims to study the effect of artesunate loaded nanofiber and its combination with spiramycin as an alternative treatment for murine toxoplasmosis. Two groups of male Swiss albino male mice were used for acute and chronic phases of Toxoplasma infection. Mice were infected with 100 viable tachyzoites of RH type-1 virulent strain intraperitoneally and orally with 100 viable tissue cysts of ME 49 Toxoplasma strain. They were orally treated with the drugs. The doses of the drug were administered from the first to the fifth day post-infection in acute infection and after six weeks in chronic one. Tachyzoites in peritoneum and cysts burden in the brain were subjected for parasitological analysis, Histopathological study and ultrastructural study by Scanning Electron microscopy (SEM). Liver enzymes analysis was done for biochemical assessment drugs. The combination artesunate loaded nanofiber and spiramycin gave the least number of tachyzoites (139) and brain tissue cysts (125) with the highest percentage reductions (85%, 84.3%) consequently. SEM revealed a reduction in the size of tachyzoites with surface irregularity and abnormal protrusions in the treated group with the drug combination. Brain tissue cyst of the same group showed a distortion in the shape and size with patches all over the surface.