A miniaturized Ames mutagenicity assay employing bioluminescent strains of Salmonella typhimurium.
Increased awareness of the role of environmental factors in carcinogenesis has led to an emphasis on preventing or minimizing exposure to genotoxicants. This is presently promoting the development of simple, rapid, cost-effective mutagenicity screening assays. We have developed a test system based on the well-known Salmonella mutagenicity assay. The lux genes, which permit cells to emit light through bioluminescence, were introduced into Salmonella typhimurium strain TA98. These bacteria were exposed for 48 h to chemicals or complex mixtures in 48-well microplates containing an appropriate liquid medium. Cells were subsequently centrifuged and resuspended in buffer. The final postexposure revertant biomass was then estimated using a microluminometer. Replication trials confirmed methodological reproducibility. Clear dose-response relationships were obtained with the direct frameshift mutagens 4NQO and 2NF. Mutagenicity threshold effect concentrations found for these compounds were comparable to those reported in the literature. Industrial effluents and environmental extracts (effluents, suspended solids) were also tested and results compared well with those of the SOS Chromotest. While further validation of this new adaptation of the Ames test will be required, it appears at this time that it could be well suited for routine screening of xenobiotics and environmental samples.
Côté
C
C
Ecole Polytechnique de Montréal, Québec, Canada.
Blaise
C
C
Delisle
C E
CE
Meighen
E A
EA
Hansen
P D
PD
D015112:4-Nitroquinoline-1-oxide / Q000633:toxicity; D002274:Carcinogens, Environmental / Q000633:toxicity; D005449:Fluorenes / Q000633:toxicity; D005798:Genes, Bacterial; D007220:Industrial Waste / Q000009:adverse effects; D008156:Luciferases / Q000235:genetics; D008163:Luminescent Measurements; D009152:Mutagenicity Tests / Q000379:methods*; D009153:Mutagens / Q000633:toxicity; D015203:Reproducibility of Results; D012486:Salmonella typhimurium / Q000201:enzymology / Q000235:genetics*
D002274:Carcinogens, Environmental; D005449:Fluorenes; D007220:Industrial Waste; D009153:Mutagens; C019499:2-nitrofluorene; D015112:4-Nitroquinoline-1-oxide; D008156:Luciferases