A sensitive, rapid ferricyanide-mediated toxicity bioassay developed using Escherichia coli |
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Authors: | Kylie Catterall David Robertson Peter R. Teasdale David T. Welsh |
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Affiliation: | a Gold Coast Water, PO Box 5042, Gold Coast Mail Centre, QLD 9729, Australia b School of Environment, Griffith University Gold Coast Campus, QLD 4222, Australia c Environmental Futures Centre, Griffith University, QLD 4222, Australia |
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Abstract: | A need for rapid toxicity techniques has seen recent research into developing new microbiological assays and characterising their toxicity responses using a range of substances. A microbiological bioassay that determines changes in ferricyanide-mediated respiration for toxicity measurement (FM-TOX) shows particular promise. The development and optimisation of an improved FM-TOX method, incorporating novel features, is described using Escherichia coli as the biocatalyst. Omission of an exogenous carbon source, used in previously described FM-TOX assays, substantially improves the assay sensitivity. In addition, the development of a two-step procedure (toxicant exposure followed by determination of microbial respiratory activity) was found to enhance the inhibition of E. coli by 3,5-dichlorophenol and four other toxicants, compared to single-step procedures. Other assay parameters, such as the ferricyanide concentration, exposure times and optical density of the biocatalyst were also optimised, sometimes based on practical aspects. Toxicity tests were carried out using the adopted technique on both organic and inorganic toxicants, with classic sigmoid-shaped dose-response curves observed, as well as some non-standard responses. IC50 data is presented for a number of common toxicants. The optimised assay provides a good foundation for further toxicity testing using E. coli, as well as the potential for expanding the technique to utilise other bacteria with complementary toxicity responses, thereby allowing use of the assay in a range of applications. |
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Keywords: | BOD, biochemical oxygen demand FM-BOD, ferricyanide-mediated biochemical oxygen demand FM-RES, ferricyanide-mediated respiration FM-TOX, ferricyanide-mediated toxicity GGA, glucose glutamic acid IC, inhibitory concentration IQ, inhibition quotient OECD, organisation for economic co-operation and development PB, phosphate buffer rcf, relative centrifugal force UNSW, University of New South Wales |
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