Vitellogenin-like protein measurement in caged Gammarus fossarum males as a biomarker of endocrine disruptor exposure: Inconclusive experience

International audienceA vitellogenin (Vg) mass spectrometry-based assay was recently developed to actively biomonitor and assess the exposure of the amphipod Gammarus fossarum to endocrine-disrupting chemicals in freshwater hydrosystems. This paper focuses on the appropriate use of this biomarker, which requires good knowledge of its basal level in males and its natural variability related to intrinsic biotic and environmental abiotic factors. To obtain the lowest biomarker variability, we first studied some of these confounding factors. We observed that the spermatogenesis stage did not have an impact on the Vg level, allowing flexibility in the choice of transplanted gammarids. In the second part of the study, males were transplanted in two clean stations for 21 days, with results indicating a spatial and temporal variability of Vg levels. These Vg changes could not be correlated to environmental factors (e.g., temperature, pH and hardness of waters). Vg induction was then assessed in 21 stations having various levels of contamination. Inductions were observed for only two of the impacted stations studied. Under reference and contaminated conditions, a high interindividual variability of Vg levels was observed in caged organisms, severely limiting the sensitivity of the biomarker and its ability to detect a significant endocrine-disruptor effect. This may be explained by unidentified environmental factors that should later be determined to improved the use of Vg as a biomarker in male G. fossarum. Moreover, as discussed in this paper, recent advancements regarding the pleiotropic functions of the Vg gene in some species may complicate the application of this biomarker in males of invertebrate specie

In Situ Reproductive Bioassay with Caged Gammarus fossarum (Crustacea): Part 2—Evaluating the Relevance of Using a Molt Cycle Temperature‐Dependent Model as a Reference to Assess Toxicity in Freshwater Monitoring

International audienceActive biomonitoring approaches are now recognized as relevant for monitoring water contamination and toxicity. Nevertheless, due to the confounding influence of variable and uncontrolled environmental conditions such as temperature, biological markers measured on transplanted individuals to assess water quality are difficult to interpret. The purpose of the present study is to propose a methodology for adapting a laboratory test of chronic sublethal toxicity based on the molting cycle of Gammarus fossarum to in situ assays. To this end, we 1) adapted the molt cycle temperature-dependent model developed in Part 1 (Chaumot et al. 2020, this issue) to the fluctuating temperatures measured in the field; 2) assessed the predictive power of our approach as a "reference value" from gammarids caged in 9 nonimpacted sites at different seasons; and 3) tested the relevance of our tool to interpret in situ reproductive bioassays from 5 upstream/downstream studies and a large-scale deployment in 12 sites. Our approach based on modeling the progress of gammarid molting cycle as a function of temperature appeared to be a relevant and robust tool for interpreting in situ observations in different environmental contexts in time and space. By avoiding using a "reference" or upstream situation as a baseline from which water quality could be assessed, this approach provides a real added value to water quality diagnosis in biomonitoring programs. Environ Toxicol Chem 2020;00:1-14. (c) 2019 SETA

Vitellogenin-like Proteins among Invertebrate Species Diversity: Potential of Proteomic Mass Spectrometry for Biomarker Development

Cost-effective methodologies along with cross-species applicability constitute key points for biomarker development in ecotoxicology. With the advent of cheaper affordable genomic techniques and high throughput sequencing, omics tools could facilitate the assessment of effects of environmental contaminants for all taxa biodiversity. We assessed the potential of absolute quantification of proteins using mass spectrometry to develop vitellogenin­(Vg)-like protein assays for invertebrates. We used available sequences in public databases to rapidly identify Vg-proteotypic peptides in seven species from different main taxa of protostome invertebrates (mollusk bivalves, crustacean amphipods, branchiopods, copepods and isopods, and insect diptera). Functional validation was performed by comparing proteomic signals from reproductive female tissue samples and negative controls (male or juvenile tissues). In a second part, we demonstrate in gammarids, daphnids, drosophilids, and gastropods that the assay validated in Vg-sequenced species can be applied to Vg-unsequenced species thanks to the evolutionary conservation of Vg-proteotypic peptide motifs. Finally, we discuss the relevance of mass spectrometry for biomarker development (specific measurement, rapid development, transferability across species). Our study supplies an illustration of the promising strategy to address the challenge of biodiversity in ecotoxicology, which consists in employing omics tools from comparative and evolutionary perspectives