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A new approach to toxicity testing in Daphnia magna: application of high throughput FT-ICR mass spectrometry metabolomics

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Abstract

Currently there is a surge of interest in exploiting toxicogenomics to screen the toxicity of chemicals, enabling rapid and accurate categorisation into classes of defined mode-of-action (MOA), and prioritising chemicals for further testing. Direct infusion FT-ICR mass spectrometry-based metabolomics can provide a sensitive and unbiased analysis of metabolites in only 15 mins and therefore has considerable potential for chemical screening. The water flea, Daphnia magna, is an OECD test species and is utilised internationally for toxicity testing. However, no metabolomics studies of this species have been reported. Here we optimised and evaluated the effectiveness of FT-ICR mass spectrometry metabolomics for toxicity testing in D. magna. We confirmed that high-quality mass spectra can be recorded from as few as 30 neonates (<24 h old; 224 μg dry mass) or a single adult daphnid (301 μg dry mass). An OECD 24 h acute toxicity test was conducted with neonates at copper concentrations of 0, 5, 10, 25, 50 μg l−1. A total of 5447 unique peaks were detected reproducibly, of which 4768 were assigned at least one empirical formula and 1017 were putatively identified based upon accurate mass measurements. Significant copper-induced changes to the daphnid metabolome, consistent with the documented MOA of copper, were detected thereby validating the approach. In addition, N-acetylspermidine was putatively identified as a novel biomarker of copper toxicity. Collectively, our results highlight the excellent sensitivity, reproducibility and mass accuracy of FT-ICR mass spectrometry, and provide strong evidence for its applicability to high-throughput screening of chemical toxicity in D. magna.

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Acknowledgements

We thank Dr. Amanda Callaghan, Prof. Richard Sibly, Dr. Lars Heckmann and Chris Hill (University of Reading) for help with establishing the D. magna culturing facility at Birmingham, Rhiannon David for assistance with the culturing, and Richard Green for valuable discussions on the mechanisms of copper toxicity. NST and MRV thank the NERC for a PhD studentship (NER/S/A/2006/14053) and an Advanced Fellowship (NER/J/S/2002/00618), respectively. RJMW acknowledges the financial support of the Darwin Trust of Edinburgh. ADS thanks the BBSRC and the Centre for Environment, Fisheries & Aquaculture Science for a PhD CASE studentship (under Cefas Seedcorn contract DP195), and TGP and TNA thank the EPSRC for a PhD studentship and DTA support.

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  1. School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK

    Nadine S. Taylor, Andrew D. Southam & Mark R. Viant

  2. Centre for Systems Biology, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK

    Ralf J. M. Weber, Olga Hrydziuszko & Mark R. Viant

  3. School of Electronic, Electrical & Computer Engineering, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK

    Tristan G. Payne & Theodoros N. Arvanitis

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  1. Nadine S. Taylor
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Taylor, N.S., Weber, R.J.M., Southam, A.D. et al. A new approach to toxicity testing in Daphnia magna: application of high throughput FT-ICR mass spectrometry metabolomics . Metabolomics 5, 44–58 (2009). https://doi.org/10.1007/s11306-008-0133-3

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