Abstract: Cuscuta epithymum Murr. (Convolvulaceae) is found in Bangladesh and has been used in traditional medicine. The present study was designed to investigate the preliminary phytochemical screening, cytotoxicity and antibacterial activities of the methanol extract of the plant stem. The preliminary phytochemical analysis was performed on the basis of the standard procedures and thin layer chromatography to confirm the presence of chemical compounds in the methanol extract of the plant. The minimum inhibitory concentration and cytotoxic activities of the extract were carried out according to broth dilution assay and brine shrimp lethality bioassay, respectively. The results of preliminary phytochemical analysis showed the presence of flavonoid, glycoside, alkaloids, carbohydrates, saponins and steroids in the plant extract. In this study, the methanol extract was subjected to Thin Layer Chromatography (TLC) in order to detect the phytoconstituents present in the extract and the best result of TLC was obtained from the solvent system comprising of toluene and ethyl acetate at the ratio of 5.5:4.5 which identified the maximum number of spots. Moreover, the extract showed moderate antibacterial activities against Bacillus megaterium, Pseudomonas aeruginosa, Escherichia coli and Salmonella typhi with MIC values of 4.96±0.20, 3.03±0.16, 3.47±0.20 and 4.07±0.08 mg mL-1, respectively. The methanol extract showed lethality against brine shrimp nauplii (LC50: 36.31 μg mL-1 and LC90: 83.18 μg mL-1 ). The results indicated that the methanol extract of the stem of the plant possessed antibacterial and cytotoxic properties with several numbers of chemical compounds.