Background: Cryptosporidium parvum is a dangerous intestinal pathogen due to its devastating effect on immunocompromised individuals. Considering low efficacy, high toxicity in addition to the development of resistance for the drugs used, this study aimed to find a new alternative treatment having the advantage of lower doses and minimal toxicity. We used a novel combination between artesunate loaded polymeric nanofiber (ALPN) and nanazoxide that had not been tried yet. Methods: Sixty Swiss Albino mice aged 6–7 wk, weighting 20-24 gm were used in Theodor Bilharz Research Institute (TBRI) Cairo, Egypt in 2017. C. parvum oocysts collected from patients were identified by polymerase chain reaction to be used for infecting animals. The effect of combination between ALPN and nanazoxide were assessed by oocyst count in stool of experimental animals using modified Ziehl- Neelsen stain and histopathological changes in intestinal tissue. Antioxidant activity of nanofiber-loaded artesunate was estimated in serum, renal, hepatic and intestinal tissues by demonstrating the reactive oxygen species and the total antioxidant capacity. It was confirmed by detection of inducible nitric oxide synthase (iNOS) antibody. Results: The novel combination between ALPN and nanazoxidehas a harmonizing effect in reducing oocyst shedding (94.4%), the mean value of the antioxidant levels in liver, intestine, kidney, and serum were the highest level (10.15, 22.4, 6.22, 14.08 respectively) resulting in the decrease of oxidative stress in tissues. Marked improvement of histopathological features was obtained. Conclusion: This combination has a promising therapeutic effect against cryptosporidiosis particularly in immunocompromised individuals considering minor toxicity.