Size-dependent effects of low level cadmium and zinc exposure on the metabolome of the Asian clam

The toxic effects of low level metal contamination in sediments are currently poorly understood. We exposed different sized Asian clams, Corbicula fluminea, to sediment spiked with environmentally relevant concentrations of either zinc, cadmium or a zinc-cadmium mixture for one week. This freshwater bivalve is well suited for sediment toxicity tests as it lives partly buried in the sediment and utilises sediment particles as a food resource. After one week, the whole tissue composition of low molecular weight metabolites was analysed by nuclear magnetic resonance spectroscopy (NMR) and gas chromatography-mass spectrometry (GC-MS). The condition index (ratio of tissue dry weight to volume inside the shell valves) was also measured. Small and large clams were clearly differentiated by their metabolic composition and the two size classes showed opposite responses to the mixture spiked sediment. No effects of zinc alone on the metabolome were found and cadmium only influenced the smaller size class. The main perturbations were seen in amino acid and energy metabolism, with small clams using amino acids as an energy resource and larger clams primarily drawing on their larger storage reserves of carbohydrates. Our study demonstrates that metabolomics is a useful technique to test for low level toxicity which does not manifest in mortality or condition index changes. The differing effects between the two size classes stress that it is important to consider age/size when conducting metabolomic and ecotoxicology assessments, since testing for the effects on only one size class makes it more difficult to extrapolate laboratory results to the natural environment