BaciUlus thuringiensis was isolated from dried tobacco residues and dead tobacco beetles (Lasioderma serricorne (F.); Coleoptera: Anobiidae) collected in a large number of locations worldwide. Eighty-eight samples of stored tobacco were analyzed and yielded 78 B. thuringiensis strains which were characterized on the basis of parasporal crystal morphology, sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles, and the results of an immunoblot analysis of the insecticidal crystal proteins. Flagellar antigen identification was used to differentiate selected isolates. Strains that produced rhomboidal crystals associated with the Coleoptera-specific pathotype (Cry Ill group) were the most abundant strains (59%o of the isolates). Preliminary toxicity assays were performed with L. serricorne larvae, and the results suggested that activity is not restricted to isolates related to the Coleoptera-specific group. The results of our survey indicate that B. thuringiensis is part of the natural microflora in the stored-tobacco environment and that this special habitat represents a source of B. thuringiensis isolates that may be used to control stored-product pests.

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A worldwide survey was conducted to evaluate the frequency and distribution of Bacillus thuringiensis populations on cured tobacco leaves during post-harvest storage. In total, 133 tobacco samples of different types and origins were analyzed. Nine percent of the samples showed the presence of B. thuringiensis, and 24 B. thuringiensis strains were isolated and characterized. The majority of the isolates produced bipyramidal crystals, and three fourths of them showed a second type of crystal protein (cuboidal or heterogeneous crystals). Only three isolates showed the rhomboidal crystal morphology characteristic of the anti-coleopteran B. thuringiensis subsp. tenebrionis. PCR analysis with primers specific for cry1 and cry3 genes revealed eight distinct cry gene profiles. The results of this study indicate that B. thuringiensis is naturally present at low frequency on the phylloplane of cured tobacco leaves and that its distribution is worldwide.

Among the insect species causing infestations and serious damages to stored commodities, the cigarette beetle, Lasiodermaserricorne (F.) and the tobacco moth, Ephestiaelutella (Hübner) are the major pests of both raw and manufactured tobacco. Post-harvest tobacco control is achieved through sanitation, insect monitoring, and fumigation with phosphine. However, insect resistance to phosphine and control failures have been reported, and increasing regulatory pressure is being exerted on fumigants. Biological control agents such as Bacillus thuringiensis (Bt) appear to be environmentally sound and potentially viable alternatives to chemical control. Bt is a bacterium that produces insecticidal crystal proteins during the sporulation phase and has been, for more than 40 years, the microorganism of choice for the biocontrol of phytophagous insect pests. It produces insecticidal crystal proteins that display specific activity against certain orders of insects and become active upon ingestion by the insect. Our laboratory has conducted extensive research and worldwide surveys to evaluate the presence of Bt in stored tobacco and has confirmed previous findings indicating that Bt may be considered part of the naturally occurring phylloplanemicroflora. Several Bt strains were isolated from tobacco and characterized by DNA and protein profiling. The insecticidal activity of selected strains and of two commercial products against the larvae of L. serricorne was determined by diet incorporation assays. Moreover, the stability of Bt spores and crystal proteins on cured tobacco leaves was assessed over a storage period of time of 30 months. Cigarette prototypes were made with Bt-treated tobacco. Standard cigarette and smoke evaluations did not show any significant difference between the test and control cigarettes. Although the tested Bt strains and products did not yield satisfactory levels of mortality at the required times and doses, the experimental results summarized in the present review indicate that B. thuringiensishas potential for the control of the cigarette beetle. The integration of conventional control approaches with novel systems based on biological agents with different modes of action should offer new avenues for the effective management of stored tobacco pests in line with integrated pest management (IPM) concepts.