A modified Ames assay reveals the mutagenicity of native cigarette mainstream smoke and its gas vapour phase

The evaluation of the mutagenic activity of cigarette smoke is mostly based on studies with condensates or extracts in the standard Ames assay. These samples only insufficiently reflect the composition of the actual generated aerosol. Therefore, such atmospheres should be analysed in their native composition to gain a real signal of its mutagenic capacity. Based on the technical difficulties of testing native air contaminants, there are no accepted methods for effective exposure of bacteria under such conditions. Therefore, we established a new experimental approach for direct exposure of bacteria in a modified CULTEX® system (Patent no. DE 19801763/PCT/EP99/00295) connected to a smoking machine. This allowed us to investigate the mutagenic activity of native mainstream smoke of the research cigarette K2R4F by exposure of Salmonella Typhimurium strains. In comparison to studies using the plate incorporation assay, the direct exposure of bacteria to smoke on the agar surface enhances contact to the aerosols. By using this modification of the Ames assay, we demonstrate that it is possible to analyse the effects of native whole smoke and the gas vapour phase of cigarettes directly and achieve a dose-dependent induction of revertants. In a number of experiments, the treatment of strains TA98 and TA100 with whole smoke and the gas vapour phase of K2R4F cigarettes resulted in the induction of revertants dependent on the dilution of smoke and the number of cigarettes smoked. Our alternative procedure of exposing bacteria directly to gases and complex mixtures offers new possibilities in the field of inhalation genotoxicology for the evaluation of genotoxicity in the Ames assay.