A semi-automated, microplate version of the SOS Chromotest for the analysis of complex environmental extracts

doi:10.1016/S0165-1161(96)90237-X

Mutation Research/Environmental Mutagenesis and Related Subjects

Volume 360, Issue 1, 17 May 1996, Pages 51-74

https://doi.org/10.1016/S0165-1161(96)90237-X Get rights and content

Abstract

Environmental monitoring for genotoxicity requires that a large number of measurements be made across space and time. This requirement demands a rapid and efficient bioassay system. The SOS Chromotest is a rapid, efficient bacterial system for the detection of DNA damaging agents. Over 100 publications have described its use on a variety of samples. Relatively few studies have used the test to examine complex mixtures. Effective testing of complex samples poses a variety of problems. Although solutions have been proposed, few have validated the resulting protocol. In this work we present a semi-automated microplate version of the SOS Chromotest for the examination of complex mixtures. Experiments were conducted to determine the optimal cell concentration, exposure time, substrate conversion time and S9 enzyme concentration. The performance of the method was evaluated using 6 reference genotoxins and 3 complex mixtures. The complex mixtures examined are extracts of diesel particulate matter, urban dust and coal tar. The results obtained indicate that optimal responses often require fewer cells (∼ 5–10 × 10⁶ CFU/ml) and a longer exposure (3 h) than that recommended in the original protocol. Interfering effects of colored and turbid samples are removed using centrifugation and initial optical density readings taken 60 min after cell resuspension and lysis. The performance of the established protocol was evaluated using mitomycin C and benzo[a]pyrene results for 207 microplates and solvent control results for 293 microplates. The results indicate that the established method is accurate, sensitive and precise. Coefficient of variation on mean SOSIP values for mitomycin C and benzo[a]pyrene are <5%. Solvent control data indicate that the standard threshold for determination of a positive response (induction factor > 1.5) is excessively conservative. All liquid transfers were automated using the Biomek^TM automated laboratory workstation. Automation permits a throughput of up to 72 samples per day and maintains excellent precision and accuracy.

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^☆: A joint contribution of The St. Lawrence Center, Environment Canada and the Department of Biology, McGill University.

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A semi-automated, microplate version of the SOS Chromotest for the analysis of complex environmental extracts☆

Abstract

Phytochemistry

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Aquat. Toxicol.

Mutation Res.

Zentralbl. Mikrobiol.

Zentralbl. Mikrobiol.

Mutation Res.

Fd. Chem. Toxicol.

Wat. Res.

Chemosphere

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Sci. Tot. Environ.

J. Biol. Chem.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Mutation Res.

Chemosphere

Mutation Res.

Mutation Res.

Mutation Res.

Carcinogens are mutagens: a simple test system combining liver homogenates for activation and bacteria for detection

Proc. Natl. Acad. Sci. USA

Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test

Mutation Res.

The prospects for a simplified and internationally harmonized approach to the detection of possible human carcinogens and mutagens

Mutagenesis

Nickel effects on phosphate uptake, alkaline phosphatase, and ATPase of a cyanobacterium

Bull. Environ. Contam. Toxicol.

Genotoxic activity of some mycotoxins using the SOS Chromotest

Mycopathologia

Biological testing-Development, application and trends in Canadian environmental protection

Toxicol. Assess.

Testing human faecal extracts for genotoxic activity with the SOS Chromotest: the importance of controlling for faecal enzyme activity

Mutagenesis

Genotoxicity studies in semiconductor industry. 1. In vitro mutagenicity and genotoxicity studies of waste samples resulting from plasma etching

J. Toxicol. Environ. Health

Modifying the SOS Chromotest for complex mixtures: solvent effects

Environ. Mol. Mutagen.

Lactose genes fused to exogenous promotors in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences

Proc. Natl. Acad. Sci. USA

Purification et propriétés de la β-galactosidase (lactase) d'Escherichia coli

Biochim. Biophys. Acta

Genotoxicity of organic extracts from atmospheric particles

Ann. NY Acad. Sci.

Applicability of the SOS Chromotest to detect urinary mutagenicity caused by smoking

Mutagenesis