Conditions for the optimal use of the L-arabinose-resistance mutagenesis test with Salmonella typhimurium

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Abstract
Different conditions of mutagenesis have been compared in order to optimize the use of the L-arabinose resistant test with Salmonella typhimurium. The mutagenesis protocols compared were the plate-incorporation, the pre-incubation and the liquid tests. Fourteen chemicals were used in the comparison: six direct-acting mutagens and eight pre-mutagens. Five concentrations of S9 (3, 7.5, 10, 15 and 33% v/v) were compared in the liquid test with pre-mutagens, and three densities of bacteria (ranging from 108 to 106 cells) were used in the comparison between the plate-incorporation and the pre-incubation mutagenesis test. In general, the liquid test proved the most sensitive mutagenesis protocol. When carrying out this test in a mass screening of mutagens, we propose to select the L-arabinose-resistant mutants in plates supplemented with 0.5 mg of D-glucose, and to express the mutagenic response as the absolute number of induced mutants. The plate-incorporation and the pre-incubation mutagenesis protocols could be considered as alternative procedures in the case of previous negative results with the liquid test. Two recommendations can be finally made in order to avoid false negative results: (a) a large population of bacteria (ranging from 108 to 107 cells) must be exposed to the mutagens in both the plate-incorporation and the pre-incubation mutagenesis tests, and (b) ideally, several S9 concentrations (ranging from 3 to 30% v/v) would be employed for testing in liquid samples of unknown mutagenicity.

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