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The in vitro susceptibilities of the isolates strain of Leishmania donovani (EMA, EAO, ZAA, HIH, MYIO and HYEA) to Sodium stibogluconate (SSG) and the experimental crude extract of Ambrosia maritime L. (Asteraceae) were determined for extracellular log-phase promastigotes and interacellular amastigotes in peripheral blood mononuclear cells (PBMCs). Susceptibility to reference drug (Pentostam) with different concentrations (1000, 500, 250, 125, 62.5 and 31.25) showed an increase in growth inhibition with the reduction in the concentration till then decrease in growth inhibition with the more reduction in concentration. While the Ambrosia maritima crude extract showed the decrease of growth inhibition with the increasing in concentration after 24, 48 and 72 hours. Ethanolic crude extract of A. maritime, it caused growth inhibition of L. donovani promastigotes at (1000, 500, 250, 125, 61.5, 31.25 mg/ mL), but the concentration 500ml/ml of the extract showed high biological activity 100% the study indicated that further analysis still to be done on the active crudes; bio-guided fractionation should also be conducted and may lead to the isolation of the major components in the active crude. Higher potency was observed against intercellular amastigots stage (IC₅₀ is different for all parasites isolates EMA=1.339 to.541 EAO=1.473 to1.719, ZAA=1.447 to 1.719, HIH=1.465 to 1.686, MYIO= 1.280 to 1.559 and HYEA=1.473 to 1.707 mg/mL), a similar was observed for reference drug (SSG) (IC₅₀ =1.945 to2.788mg/mL).