The aim of this work was to investigate and compare the phytochemical screening and antimicrobial activities of different crude extracts from dry and fresh leaves of Datura metel. Different organic solvents including methanol, chloroform, hexane, ethyl acetate and butanol were used to prepare the crude extracts from the fresh and dry leaves. Antimicrobial activities of different crude extracts from dry and fresh leaves of Datura metel were determined by agar disc diffusion method with minor modification. In vitro phytochemical screening for all crude extracts from both dry and fresh leaves was tested and shown positive result for alkaloid, flavonoid, saponin and tannin compounds. However, all the crude extracts did not show positive results for steroids and triterpenoid compounds. The methanol crude extract and its derived fractions from dry and fresh leaves showed small and moderate antibacterial potential with one gram positive (Staphylococcus aureus) and three gram negative (Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa) bacteria in the range of 0–17%. In conclusion, all organic crude extracts from both fresh and dry leaves could be used as potential sources of new antimicrobial properties.