Bacterial lipases are a highly versatile class of hydrolytic enzymes which catalyze the hydrolysis of triglycerol to glycerol and fatty acids. They play an important role in biotechnological and industrial processes, including oleo chemical, food, pharmaceutical detergent formulation, cosmetic, leather, textile, and paper industries applications. Tremendous interest among scientists and industrialists has been arisen in the last decade in microbial lipases due to their versatility and ease of mass production.In the current work, natural oils such as Triglyceride, Tween 20, Tween 80, Olive oil, Sweet almond, Celery oil, Sesame, Rosemary oils were implemented to enhance lipase production by soil and pathogenic bacterial isolates. Results revealed that triglyceride and tween 80 had the best enhancing activity for lipase production in most isolates. It was also shown that Pseudomonas and Bacillus were the most potential lipase producing bacteria.A novel modified tributary agar was prepared by the addition of Sudan blue dye and used for the detection of lipase producers. It was shown to give a very clear zone even in the case of weak producing strains as compared to the original medium.Specific lipase activity had been determined Staphylococcus aureus (in the present research. The highest lipase activity was demonstrated by Pseudomonas (4.5u/ml), while Staphylococcus aureus did not show any lipase activity.