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Bacillus thuringiensis spore forming, Gram positive aerobic bacteria whose spores consist of the crystal protein which are found to have the insecticidal activity to the specific insect orders. Bacillus thuringiensis are considered to be cosmopolitan in nature and a great difference occurs between the two strains from different habitat. The present study was conducted on the collection of samples and solation of the B.t. from these samples. These isolates were identified on the basis of microscopic techniques and staining techniques. Culturing of these isolates was made on the specific media such as T3. Culturing of these bacteria was done to get enough amount of the B.t. spore cell. Rearing of the Tribolium castaneum a flour beetle was done by understanding the physiology of the beetles. Then biotoxic effect of these bacteria was determined by the process of Biotoxicity assay of the Bacillus thuringiensis spores against Triboium castaneum in glass vials in triplicates. The sevenday repeated observation of these isolates was presented in the tabular form and LC50 is calculated by the Probit analysis programme.