show Abstracthide AbstractChemical contamination is a common threat to biota thriving in estuarine and coastal ecosystems. In particular, trace metals tend to accumulate and exert deleterious effects on small invertebrates such as zooplankton, which are essential trophic links between phytoplankton and higher-level consumers in aquatic food webs. Beyond the direct effects of the contamination, we hypothesized that metal exposure could also affect the zooplankton microbiota, which in turn might further impair host fitness. To assess this assumption, copepods (Eurytemora affinis) were sampled in the oligo-mesohaline zone of the Seine estuary and exposed to dissolved copper (25 µg.L-1) over a 72-hour time period. Copepod response to copper treatment was assessed by determining transcriptomic changes in E. affinis along with shifts in its microbiota. Unexpectedly, very few genes were differentially expressed in copper-treated copepods compared to controls, with most of the reported differences involving genes upregulated in males compared to females. In contrast, copper increased the taxonomic diversity indices of the microbiota and resulted in substantial compositional changes at both the phyla and genus levels. Phylogenetic reconstruction of the microbiota further suggested that copper mitigated phylogenetic relatedness of taxa at the basal tree structure of the phylogeny, whereas it strengthened it at the terminal branches. Increased terminal phylogenetic clustering in copper-treated copepods concurred with higher proportions of bacterial genera previously identified as copper resistant (e.g., Pseudomonas, Acinetobacter, and Alkanindiges) and a higher relative abundance of the copA gene encoding a periplasmic inducible multi-copper oxidase. Overall, these results revealed very contrasting responses of E. affinis and its microbiota to copper exposure. The enrichment in micro-organisms likely to perform copper sequestration and/or enzymatic transformation processes, underlines here the need to follow the microbial component during the evaluation of the vulnerability of the zooplankton to the metallic stress. Overall design: Comparative gene expression analysis of RNA-seq data for adult male and female copepods Eurytemora affinis exposed to copper via the water. Triplicates were made for each condition (copper and control) and sex (male and female). Each sample correspond to a pool of 30 organisms.