• Innovative Strategies for the Application of Molecular Biomarkers in Ecotoxicology
• Mass spectrometry enables rapid assessment of organism health status.
• Ecotoxicoproteomics is a relevant approach for environmental monitoring.

The routine use of biomarkers in environmental biomonitoring faces several drawbacks, especially in invertebrates. Among these limitations, the lack of validated species-specific biomarkers and multibiomarker methodologies comprise major constraints. Based on gene and protein catalogs obtained from proteogenomics experiments previously performed on the ecotoxicological-relevant species Gammarus fossarum, this thesis aimed at identifying and validating molecular biomarkers for the diagnostic of toxic perturbations in this species. Following recent methodologies for the development of biomarkers in human disease diagnosis, we implemented a fast, specific, quantitative multiplexed targeted proteomics assay (using Selected Reaction Monitoring mass spectrometry) to study dozens of protein biomarker candidates simultaneously. The assay was applied to assess the physiological importance of protein candidates, and to assess their pertinence as biomarkers after laboratory and field exposures to chemical contamination. The second part of this thesis aimed at the development of specific endocrine disruption biomarkers, through two complementary approaches. The first approach, based on a comparative shotgun proteomic analysis using a known endocrine disruptor for arthropods, allowed exhaustively increasing the whole-proteome analysis through the detection of roughly 4000 proteins (53 modulated by the exposure). The second comprised sequence homology searches, phylogenetic analyses, and gene expression studies for proposing new biomarker candidates identified from literature searches

Publications

• Charnot, A., D. Gouveia, J. Armengaud, C. Almunia, A. Chaumot, J. Lemoine, O. Geffard and A. Salvador (2017). "Multiplexed assay for protein quantitation in the invertebrate Gammarus fossarum by liquid chromatography coupled to tandem mass spectrometry." Analytical and Bioanalytical Chemistry 409(16): 3969-3991. DOI: 10.1007/s00216-017-0348-0
• Gouveia, D.; Chaumot, A.; Charnot, A.; Queau, H.; Armengaud, J.; Almunia, C.; Salvador, A.; Geffard, O., Assessing the relevance of a multiplexed methodology for proteomic biomarker measurement in the invertebrate species Gammarus fossarum: a physiological and ecotoxicological study. Aquatic Toxicology 2017, 190, 199-209. DOI:10.1016/j.aquatox.2017.07.007
• Gouveia, D.; Chaumot, A.; Charnot, A.; Almunia, C.; François, A.; Navarro, L. ; Armengaud, J.; Salvador, A.; Geffard, O., Ecotoxico-proteomics for aquatic environmental monitoring: first in situ application of a new proteomics-based multibiomarker assay using caged amphipods. Environmental Science and Technology, 2017, 51 (22), 13417-13426. DOI: 10.1021/acs.est.7b03736.
• Gouveia, D., F. Bonneton, C. Almunia, J. Armengaud, H. Quéau, D. Degli-Esposti, O. Geffard and A. Chaumot (2018). "Identification, expression, and endocrine-disruption of three ecdysone-responsive genes in the sentinel species Gammarus fossarum." Scientific Reports 8(1): 3793. DOI:10.1038/s41598-018-22235-7
• Charnot, A., D. Gouveia, S. Ayciriex, J. Lemoine, J. Armengaud, C. Almunia, A. Chaumot, O. Geffard and A. Salvador (2018). "On-Line Solid Phase Extraction Liquid Chromatography-Mass Spectrometry Method for Multiplexed Proteins Quantitation in an Ecotoxicology Test Specie: Gammarus fossarum." Journal of Applied Bioanalysis 4(3): 81-101. DOI: 10.17145/jab.18.012
• *Trapp, J., *D. Gouveia, C. Almunia, O. Pible, D. Degli Esposti, J.-C. Gaillard, A. Chaumot, O. Geffard and J. Armengaud (2018). "Digging Deeper Into the Pyriproxyfen-Response of the Amphipod Gammarus fossarum With a Next-Generation Ultra-High-Field Orbitrap Analyser: New Perspectives for Environmental Toxicoproteomics." Frontiers in Environmental Science 6(54). DOI : 10.3389/fenvs.2018.00054. *1st co-authors

Cite the thesis

Duarte Domingos Gouveia. Molecular approaches for the discovery, development and application of toxicity biomarkers in gammarids. Ecotoxicology. Université de Lyon, 2017. English. ⟨NNT : 2017LYSE1318⟩. ⟨tel-01723905⟩

Access manuscript on HAL thèses

And then?

Since 2021, Duarte is Research Associate in Translational Proteomics & Metabolomics at Evotec France.

Follow his career path on ORCID

And connect him on LinkedIn