Leishmaniasis is still health treating problem in low health literacy regions of the world. On the other hand, toxicity, side-effects and expenses of the drugs, increased demand for natural sources material. Aim: Antileishmanial activity of Yazd spirogyra spp extracts (SSE) on the viability of Leishmania (L) major promastigote (PMs) an in vitro study. Sufficient root of spirogyra spp were minced, sterilized and prepared as tropica with concentration of 5, 25,125 and 725 μg. Leishmania major [MRHO/IR/75/ER] cultured then Amastigote was transferred to screw capped vials containing 5 ml of RPMI1640 media to which different concentrations of 5, 25,125 and 725 μg SSE were added and each concentration was done in triplicates. Each run also included control. The mortality of parasitoids was measured by the enzyme-linked assay (ELISA) methods. The percentage viability of SSE and showed that viability of parasites in PMs significantly decreased in SSE 5, 25,125 and 725 μg treatment compare to control group (P=0.000, F=30.65). As shown, after 48hour, the percentage inhibition of spirogyra spp and stationary and logarithmic phases of Leishmania major [MRHO/IR/75/ER] that the percent inhibition is density in time (P=0.000, F=16.03).Then 24 h after, the Antileishmanial bioassay were determined by XTT detecting kit using chemiluminescent assay. According to the results dose dependent decrease detected on variety on Cutaneous Leishmaniasis using different levels of Glocantime and hydroalcholic extracts of green prevalent algae (P=0.000, F=24.985). Also, no significant difference detected on anti-Leishmania activity of the SSE compared to Glocantime (P>0.05). These results suggest green prevalent algae had medical potential similar to the Glocantime.