Many recent studies have shown the benefits of scorpion venom, as it components may be used as potential candidates for drug development, especially as antimicrobial compounds against both gram-negative or gram-positive bacterial strains and fungi. Therefore, the present study was conducted to evaluate the antimicrobial activity of venom from two scorpion species that inhabit Saudi Arabia (Leiurus quinquestriatus and Androctonus crassicauda) against four bacterial strains: two gram-negative bacteria (Escherichia coli and Salmonella sp.) and two gram-positive bacteria (Staphylococcus aureus and Paenibacillus larvae). Scorpions were collected from two cities (Riyadh and Taif) and the venom was extracted using electrical stimulation, and then lyophilized. The antimicrobial activity of the lyophilized venoms was evaluated in vitro by an agar well diffusion method against standard bacterial strains by measuring the inhibition zone of each bacterial strain upon treatment with scorpion venom. Our results showed that the crude venom of the two scorpion species had dose-dependent inhibitory effects on the gram-positive and gram-negative bacteria studied.