Stored tobacco is commonly attacked by insects such as Lasioderma serricorne (F.) (Coleoptera: Anobiidae). The objective of this research was to isolate and to identify strains of Bacillus thuringiensis with potential to reduce L. serricorne populations in stored tobacco. In order to reach such objective, 168 samples from the tobacco warehouses and soil were investigated. Nine strains from COLTABACO warehouses were isolated with parasporal rhomboidal and bypiramidal crystals. PCR analysis indicated that these strains contained cry1Ab, cry1Ac, cry1B y cry3A genes. SDS-PAGE and Western Blot of the Cry proteins showed that these strains contain crystal proteins of 130, 70, 67, and 30 kDa; and a positive immunological relationship to the crystal proteins of B. thuringiensis subsps. tenebrionis and kurstaki. The insecticidal activity of the spore crystal suspension indicated that these strains are active against larvae of L. serricorne and Spodoptera frugiperda. The half lethal dose obtained was in the range of 428 an 3042.3 ng/cm²; in L. serricorne mortality was between 65 to 67% with a dose of 0.1 mg/ml (diet/spore crystal). b-exotoxin production was checked by bioassays Musca domestica larvae and indicated that these do not produce this toxin. The results show that these strains could be considered as components for a integrated perst management program of L. serricorne.