An experimental study was undertaken to assess the efficiency of Pseudomonas aeruginosa, Bacillus subtilis, and Acinetobacter lwoffi isolated from petroleum contaminated water and soil samples to degrade crude oil, separately and in a mixed bacterial consortium. Capillary gas chromatography was used for testing the effect of those bacterial species on the biodegradation of crude oil. Individual bacterial cultures showed less growth and degradation than did the mixed bacterial consortium. At temperature 22°C, the mixed bacterial consortium degraded a maximum of 88.5% of Egyptian crude oil after 28 days of incubation. This was followed by 77.8% by Pseudomonas aeruginosa, 76.7% by Bacillus subtilis, and 74.3% by Acinetobacter lwoffi. The results demonstrated that the selected bacterial isolates could be effective in biodegradation of oil spills individually and showed better biodegradation abilities when they are used together in mixed consortium.