International audience | Watersheds and estuaries are impacted by multiple anthropogenic stressors that affect their biodiversity and functioning. Assessing their ecological quality has consequently remained challenging for scientists and stakeholders. In this paper, we propose a multidisciplinary approach to identify the stressors in seven small French estuaries located in agricultural watersheds. We collected data from landscape (geography, hydrobiology) to estuary (pollutant chemistry) and fish individual scales (environmental signatures, proteomics). This integrative approach focused on the whole hydrosystems, from river basins to estuaries. To characterize each watershed, we attempted to determine the land use considering geographic indicators (agricultural and urbanised surfaces) and landscape patterns (hedges density and riparian vegetation). Juveniles of European flounder (Platichthys flesus) were captured in September, after an average residence of five summer months in the estuary. Analyses of water, sediments and biota allowed to determine the concentrations of dissolved inorganic nitrogen species, pesticides and trace elements in the systems. Environmental signatures were also measured in flounder tissues. These environmental parameters were used to establish a typology of the watersheds. Furthermore, data from proteomics on fish liver were combined with environmental signatures to determine the responses of fish to stressors in their environments. Differential protein abundances highlighted a dysregulation related to the detoxification of xenobiotics (mainly pesticides) in agricultural watersheds, characterized by intensive cereal and vegetable crops and high livestock. Omics also revealed a dysregulation of proteins associated with the response to hypoxia and heat stress in some estuaries. Furthermore, we highlighted a dysregulation of proteins involved in urea cycle, immunity and metabolism of fatty acids in several systems. Finally, the combination of environmental and molecular signatures appears to be a relevant method to identify the major stressors operating within hydrosystems.

Like many urban catchments, the River Thames in London is contaminated with plastics. This pollutant is recorded on the river banks, in the benthic environment and in the water column. The present study was conducted to assess the extent of microplastic ingestion in two River Thames fish species, the European flounder (Platichthys flesus) and European smelt (Osmerus eperlanus). Samples were collected from two sites in Kent, England; Erith and Isle of Grain/Sheppey, near Sheerness, with the latter being more estuarine. The results revealed that up to 75% of sampled European flounder had plastic fibres in the gut compared with only 20% of smelt. This difference may be related to their diverse feeding behaviours: European flounder are benthic feeders whilst European smelt are pelagic predators. The fibres were predominantly red or black polyamides and other fibres included acrylic, nylon, polyethylene and polyethylene terephthalate and there was no difference in occurrence between the sites sampled.

Although the use of tributyltin in antifouling paints has been banned, this compound is still a serious pollutant of the marine environment. This paper describes a unique study in which European flounder (Platichthys flesus) were chronically (8 months) exposed to bis(tri-n-butyltin)oxide (TBTO) in the water under controlled laboratory conditions. Residue levels in selected tissues (liver, muscle) and general health status indices were measured and the effects on several organs (gills, liver, mesonephros, ovary/ testis, spleen, and gastrointestinal tract) were examined histopathologically. Additionally, morphometric analysis of the thymus was performed. The major finding is that exposure of flounder to 5 mg TBTO/l over a period of 8 months, resulting in body burdens comparable to high field levels, induced significant reduction of thymus volume, possibly affecting immunocompetence of the animals. Chronic exposure of European flounder to tributyltin is therefore likely to affect the general health status of this species in heavily polluted aquatic environments.

Watersheds and estuaries are impacted by multiple anthropogenic stressors that affect their biodiversity and functioning. Assessing their ecological quality has consequently remained challenging for scientists and stakeholders. In this paper, we propose a multidisciplinary approach to identify the stressors in seven small French estuaries located in agricultural watersheds. We collected data from landscape (geography, hydrobiology) to estuary (pollutant chemistry) and fish individual scales (environmental signatures, proteomics). This integrative approach focused on the whole hydrosystems, from river basins to estuaries. To characterize each watershed, we attempted to determine the land use considering geographic indicators (agricultural and urbanised surfaces) and landscape patterns (hedges density and riparian vegetation). Juveniles of European flounder (Platichthys flesus) were captured in September, after an average residence of five summer months in the estuary. Analyses of water, sediments and biota allowed to determine the concentrations of dissolved inorganic nitrogen species, pesticides and trace elements in the systems. Environmental signatures were also measured in flounder tissues. These environmental parameters were used to establish a typology of the watersheds. Furthermore, data from proteomics on fish liver were combined with environmental signatures to determine the responses of fish to stressors in their environments. Differential protein abundances highlighted a dysregulation related to the detoxification of xenobiotics (mainly pesticides) in agricultural watersheds, characterized by intensive cereal and vegetable crops and high livestock. Omics also revealed a dysregulation of proteins associated with the response to hypoxia and heat stress in some estuaries. Furthermore, we highlighted a dysregulation of proteins involved in urea cycle, immunity and metabolism of fatty acids in several systems. Finally, the combination of environmental and molecular signatures appears to be a relevant method to identify the major stressors operating within hydrosystems.

Frequencies of eight nuclear abnormalities (NAs) reflecting environmental genotoxicity and cytotoxicity, were examined in 739 specimens of herring (Clupea harengus), flounder (Platichthys flesus) and cod (Gadus morhua) collected between 2009 and 2017 at 50 study stations located in the Gulf of Gdansk and the southern part of the Gdansk Deep (the Baltic Sea). The highest levels of geno-cytotoxicity were recorded in fish caught at stations located in close proximity to chemical and conventional munitions dumping sites or in zones polluted by Chemical Warfare Agents (CWAs). The values of NAs were significantly higher (up to 51-fold compared with the reference level) in herring caught at seven stations and in cod from one station located close to the munitions dumping sites. Exceptionally high total genotoxicity (∑Gtox) risk was found for flounder collected from 18 (72.0%) stations, herring caught at 12 (38.7%) stations and cod caught at four (17.4%) studied stations.

Microplastic ingestion has been reported for several marine species, but the level of contamination in transitional systems and associated biota is less known. The aim of this study was to assess the occurrence of microplastic ingestion in three commercial fish species: the sea bass (Dicentrarchus labrax), the seabream (Diplodus vulgaris) and the flounder (Platichthys flesus) from the Mondego estuary (Portugal). Microplastics were extracted from the gastrointestinal tract of 120 individuals by visual inspection and digestion solution. A total of 157 particles were extracted from 38% of total fish (96% fibers), with 1.67 ± 0.27 (SD) microplastics per fish. Significantly higher amount of ingested microplastics was recorded for D. vulgaris (73%). The dominant polymers identified by μ-FTIR were polyester, polypropylene and rayon (semi-synthetic fiber). It is reported for the first time the presence of this pollutant in fish populations from the Mondego estuary raising concerns on their potential negative effects.

A multi-biomarker approach was developed to evaluate responses of European flounder (Platichthys flesus) in three contrasted estuaries over the English Channel: the Canche (pristine site), Tamar (heavy metals and PAHs contamination) and Seine (heavily pollution with a complex cocktail of contaminants). The condition factor and several biomarkers of the immune system, antioxidant enzymes, energetic metabolism and detoxification processes were investigated in young-of-the-year (0+) and one-year-old (1+) flounder. Results underlined the difference between the pristine site and the Seine estuary which showed a lower condition factor, a modulation of the immune system, a higher Cytochrome C oxidase activity, and an up-regulation of BHMT expression. The moderate biomarker responses in the Tamar fish could be linked to the specific contamination context of this estuary. Flounder life history traits were analyzed by otolith microchemistry, in order to depict how the fish use their habitat and thus respond to chemical stress in estuaries.

Platichthys flesus is often used as a sentinel species to monitor the estuarine water quality. In this study, we carried out an experimental contamination of fish using a PAHs/PCBs mixture, which was designed to mimic the concentrations found in the Seine estuary (C1) and 10 times these concentrations (C2). We used a proteomic approach to understand the molecular mechanisms implied in the response of P. flesus to these xenobiotics. We showed that 54 proteins were differentially accumulated in one or several conditions, which 34 displayed accumulation factors higher than two. 18 of these proteins were identified by MALDI TOF–TOF mass spectrometry. The results indicated the deregulation of oxidative stress- and glutathione metabolism-(GST, GPx) proteins as well as of several proteins belonging to the betaine demethylation pathway and the methionine cycle (BHMT, SHMT, SAHH), suggesting a role for these different pathways in the P. flesus response to chemical contamination.