Natural products play an important role as environmentally friendly agents that can be used against parasitic diseases. Many Eimeria species cause eimeriosis in poultry. The negative effects of synthetic anti-coccidiosis medications necessitate the quest for alternative treatments derived from medicinal plants in the treatment of eimeriosis. The study was conducted to evaluate the effects of Calotropis procera leaf extract (CPLE) (Madar) on the sporulation of Eimeria oocysts and sporozoites that affect mammalian jejunum and to obtain the best concentration for sporulation inhibition and infection prevention. Extracts were tested in vitro to prevent oocyst sporulation, wall deformity, and anti-sporozoite activity with Eimeria papillata. The plant-chemical compounds analysis of CPLE some active compounds were shown as well as CPLE in vitro effects at various concentrations (200, 100, 50, 25,12.5, and 6.25 mg/mL), while potassium dichromate solution 2.5% and Toltrazuril 25 mg/mL were administered as the control groups. C. procera leaf extract showed the highest inhibitory percentage on E. papillata oocyst at 200 mg/mL of extract, approximately 91%. In addition, CPLE showed the sporozoite highest viability inhibitory percentage on E. papillata at 200 mg/mL of extract, approximately 88%, and the lowest efficacy was 5% at 6.25 mg/mL. Also, we noticed the deformation and destruction of the oocyst wall based on the concentration rate. Sporulation inhibition rate is significantly affected by incubation time and treatment concentration ratio. The results showed that Madar has an effective, inhibitory potential, and protective effect on coccidian oocyst sporulation and sporozoites of E. papillata.