engleski

A Quick, Comprehensive and Reproducible Protocol for the Integrative Multi Stressor - Multi OMICS Analyses of Environmental Samples

Man-made contaminants have become part of our everyday lives and their presence in the environment is readily acknowledged. Due to their omnipresence in the environment, microplastics and certain chemicals such as endocrine disrupting compounds (EDCs) and pharmaceuticals (PhACs) have become contaminants of emerging concern. The general approach in (eco)toxicological studies has mainly focused on the impact of individual stressors (e.g. EDCs, PhACs or microplastics) on aquatic biota. As individual stressors in natural habitats rarely acts in isolation and many ecosystems are characterized by the cooccurrence of stressors, it has been suggested that future studies should consider multiple stressor environments. The response of biological systems to stress can range from the individual to the community level. On the individual level, stress may alter the physiology of the cellular and molecular network. Incorporating stress information encoded at the molecular level will certainly provide a more layered, complete and informative view of the processes occurring in multiple stress environments. The integration of multiple measurements (i.e. quantification of stressors, proteomics and metabolomics) may provide a more informative view on the effects of multiple stressors on aquatic organisms. The possibility of extracting a variety of molecules and performing multiple measurements on single samples is very attractive, but also challenging and a first step is to develop and validate analytical methods for their extraction and determination in biotic matrices. Our study focused on the development of a rapid and comprehensive protocol for the simultaneous extraction of emerging contaminants (EDCs, PhACs and microplastics), metabolites and proteins for an integrative multi-stressor and multi-omics approach. We based the protocol on extraction with organic solvents and solid phase extraction clean up procedures. Special focus was placed on testing the purification efficiency of complex matrices, taking into account the values of the matrix effect and recovery, and on accelerating and simplifying the process of isolation of microplastic particles from biota samples. To evaluate the broad applicability of this method, we performed extraction procedures on a variety of biotic samples. Here we present the results of the application of this protocol in a mesocosm study examining the impact of stressors on freshwater biota.

metabolites ; proteines ; microplastics

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