Because of developing resistance to the existing anthelmintic drugs, there is a need for new anthelmintic agents. Tobacco plant has alkaloid materials that have antiparasitic effect. We investigated the in vitro anthelminthic effect of aqueous and alcoholic extract of Tobacco (Nicotiana tabacum) against M. marshalli. For investigating this effect, we prepared three dilutions of aqueous and alcoholic extract of Tobacco (25, 50 and 75 mg/ml). The worms exposed to extracts for 10 h at 25-30°C. The buffer PBS used as negative control and 50 mg/ml dilution of Levamisole used as standard reference. In each group, 50 worms were examined. We used an inhibition mobility test for our study. Survival analysis with Cox proportional hazard model was used for data analysis. The result showed that compared with Levamisole 50 mg/ml, dilution of 25 and 50 mg/ml of the aqueous extract had the same anthelminthic effects (P [ 0.05), but 75 mg/ml dilution of the aqueous extract and dilution of 25, 50 and 75 mg/ml of alcoholic extract had more anthelminthic effect (P \ 0.05). Overall, extracts of Tobacco possess considerable anthelminthic activity and more potent effects were observed with the highest concentrations. Therefore, the in vivo study on Tobocco in animal models is recommended.