A metabolomics based approach to assessing the toxicity of the polyaromatic hydrocarbon pyrene to the earthworm Lumbricus rubellus

The biochemical response of the earthworm Lumbricus rubellus to pyrene exposure was assessed using 1H nuclear magnetic resonance (NMR) spectroscopy, gas chromatography mass spectrometry (GC-MS) and pattern recognition techniques. Both analytical methods enabled the establishment of reproducible metabolic profiles. NMR analysis identified a total of 32 metabolites while GC-MS identified 51. The results demonstrate that not only is pyrene toxic to L. rubellus, but that alterations in its normal metabolic profile could be observed even when individuals were exposed to concentrations of 40 mg kg-1: a pollution level that is both below the concentration previously found to significantly reduce reproduction and within the range of polycyclic aromatic hydrocarbons (PAHs) found on some contaminated sites. Pyrene was found to cause a dose dependant decrease in lactate and the concentrations of the saturated fatty acids tetradecanoic, hexadecanoic and octadecanoic acid and an increase in production of the amino acids alanine, leucine, valine, isoleucine, lysine, tyrosine and methionine. It is proposed that this indicates impaired glucose metabolism, with an associated increase in fatty acid metabolism and changes in TCA cycle intermediates. This study demonstrates the versatility of metabolomics as a tool to monitor toxicity in the environment as opposed to utilising model species studied in a laboratory setting. Since it is a non-carcinogenic PAH, we propose that the metabolic changes observed in worms may reflect the non-specific toxic effects of pyrene as a typical, non-polar organic compound.