Effect of ALE and ASE on intra-macrophagic amastigotes. (a). Estimation of anti-amastigote efficacy. Parasitized RAW 264.7 macrophages were cultured with ALE and ASE at different concentrations and percent reduction in amastigote growth was determined as described in Methods. ***P < 0.001in comparison to infected control. (b). Microscopic imaging of L. donovani infected macrophages. Images depict giemsa stained intracellular amastigote forms residing in RAW macrophages. Arrow indicates the internalized parasites, where a decrease in amastigote number in treated samples is indicative of their anti-amastigote potential. (c). Cytotoxicity of ALE and ASE against mammalian macrophages. RAW 264.7 cells (2 × 106 ml−1) were incubated with ALE and ASE (0 to 500 μg ml−1) and cell viability was ascertained by MTT assay. Data shown are from one of the three independent experiments and are expressed as mean ± SEM of samples in triplicates. *P < 0.05, #P < 0.01 with respect to macrophage control without any treatment.