Anchoring metabolic changes to phenotypic effects in the chlorophyte Scenedesmus vacuolatus under chemical exposure

In order to derive a causal understanding of toxic effects in organisms, ecotoxicology may benefit from linking molecular changes, evaluated by 'omics'-techniques, to phenotypic observations. However, an approach to link these observation levels is still lacking. The aim of this study was to relate metabolic changes in the chlorophyte Scenedesmus vacuolatus to established parameters of toxicity. Therefore, synchronized cultures of the alga were exposed for 14 h to the phytotoxicant N-phenyl-2-naphthylamine (PNA) in the range of 0.00089 μmol L⁻¹ (environmental concentrations) up to 1.82 μmol L⁻¹. Cell growth and photosynthesis inhibition were evaluated but revealed no effect of PNA at experimental concentrations below 0.456 μmol L⁻¹. Changes in the biochemical composition of algae were measured by GC-MS in both polar and non-polar phases. PCA uncovered no separation in the multivariate pattern of mass spectral features at exposure concentrations below 0.00356 μmol L⁻¹ of PNA. However, a clear separation was detected at concentrations higher than 0.00713 μmol L⁻¹. A combined visualization of PCA results for metabolic changes and concentration-response relationships for growth and photosynthesis inhibition revealed (I) a two orders of magnitude higher sensitivity of metabolomics to detects changes after PNA exposure compared to the phenotypic parameters measured and (II) two types of metabolic responses: one group of features was reflecting pharmacological effects at low exposure concentrations and the second group corresponded to adverse effects along with conventional observations of toxicity.