Abstract
Environmental metabolomics is a powerful approach to investigate the response of organisms to contaminant exposure at a molecular scale. However, metabolomic responses to realistic environmental conditions can be hindered by factors intrinsic to the environment and the organism. Hence, a well-designed experimental exposure associated with adequate statistical analysis could be helpful to better characterize and relate the observed variability to its different origins. In the current study, we applied a multifactorial experiment combined to Analysis of variance Multiblock Orthogonal Partial Least Squares (AMOPLS), to assess the metabolic response of wild marine mussels, Mytilus galloprovincialis, exposed to a wastewater treatment plant effluent, considering gender as an experimental factor. First, the total observed variability was decomposed to highlight the contribution of each effect related to the experimental factors. Both the exposure and the interaction gender × exposure had a statistically significant impact on the observed metabolic alteration. Then, these metabolic patterns were further characterized by analyzing the individual variable contributions to each effect. A main change in glycerophospholipid levels was highlighted in both males and females as a common response, possibly caused by oxidative stress, which could lead to reproductive disorders, whereas metabolic alterations in some polar lipids and kynurenine pathway were rather gender-specific. This may indicate a disturbance in the energy metabolism and immune system only in males. Finally, AMOPLS is a useful tool facilitating the interpretation of complex metabolomic data and is expected to have a broad application in the field of ecotoxicology.
Highlights
Coastal areas around the world are subjected to multiple pressures, including an increase of submarine sewage outfalls
We propose to apply the AMOPLS method for the first time in environmental metabolomics in order to (i) bring out metabolites modulated in the same way in male and female mussels, and (ii) gain insight into how gender influences responses to exposure
liquid chromatography—high resolution mass spectrometry (LC-HRMS) analysis of digestive glands collected from mussels exposed either to solvent control (SC, n = 9 males and n = 10 females) or wastewater treatment plants (WWTPs) effluent extract (n = 10 males and n = 10 females) was carried out in both ESI+ and ESI− modes
Summary
Coastal areas around the world are subjected to multiple pressures, including an increase of submarine sewage outfalls. Approaches helping to characterize the response of marine organisms to such multi-contamination without preconception are key for understanding the impact of WWTP effluents on coastal areas. Environmental metabolomics has been commonly and efficiently applied to investigate more deeply mechanisms and modes of action of single contaminants or mixtures [9,10,11]. This approach is based on the identification of low molecular weight metabolites (50–1500 Da) whose production and levels vary with the physiological, developmental, or pathological state of cells, tissues, organs, or whole organisms [10,12]
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