peer reviewed | The northwestern shelf of the Black Sea has been affected by eutrophication and bottom hypoxia since the sixties. Consequently, the macrozoobenthos has suffered a well-established decline in biodiversity. However, the nature of the current benthic communities remains questionable. From 1995 to 2017, we compiled species and abiotic data for 138 sites over the shelf. Through an appropriate multivariate analytical approach, we identified benthic community changes solely due to organic pollution variations. Our results show signs of recovery with an increase in biodiversity and proportion of species vulnerable to organic enrichment. These changes were related to a decrease in riverine loads and subsequent eutrophication. However, some long-lived species typical of the area still did not exhibit noticeable recovery, which suggests that either the recovery process has not yet been achieved or some environmental conditions are still not met to warrant a sea floor ecosystem state substantially healthy. | 14. Life below water

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.

Hypoxia (low oxygen) often occurs in aquatic ecosystems that receive effluent from municipal wastewater treatment plants (WWTP). The combination of hypoxia and WWTP effluent could impair fish health, because WWTP effluent contains multiple contaminants that could disrupt the physiological pathways fish use to cope with hypoxia, but the interactive effects of these stressors on fish physiology are poorly understood. We have examined this issue by exposing mummichog killifish (Fundulus heteroclitus) to hypoxia (5 and 2 kPa O₂) and/or 100% WWTP effluent for 21 days in a full factorial design. We then measured hypoxia tolerance, whole-animal metabolism, gill morphology, haematology, and tissue metabolites. In clean water, killifish responded to chronic hypoxia with improvements in hypoxia tolerance, as reflected by increases in time to loss of equilibrium at 0.5 kPa (tLOE). These improvements occurred in association with increases in the exposed surface of gill lamellae that resulted from a regression of interlamellar cell mass (ILCM). Concurrent exposure to wastewater attenuated the increases in tLOE and gill remodeling in chronic hypoxia, and nearly depleted brain glycogen stores. Therefore, exposure to WWTP effluent can disrupt the physiological mechanisms fish use to cope with chronic hypoxia and impair hypoxia tolerance. Our research suggests that the combination of stressors near WWTPs can have interactive effects on the physiology and health of fish.

Nitrogen (N) pollution and the resulting eutrophication can have deleterious consequences on estuaries, such as hypoxia, fish kills, and loss of biotic diversity. An understanding of N sources and cycling in estuaries is fundamental to determining how to effectively manage these ecologically and commercially important areas. We applied a multiple-isotopic approach to examine the transformations and sources of the N pools in the Pearl River Estuary (PRE) during winter. The surface water in the West PRE was characterized by low salinity and high NO₃⁻, while that in the east had high salinity and low NO₃⁻. The NO₃⁻ in the West PRE was largely regulated by a conservative mixing process. In contrast, assimilation and nitrification dominated in the East PRE, which was attributed to the long water-residence time. For the first time, the source contributions of NO₃⁻ and NH₄⁺ were estimated by isotope mixing models. Our results suggest that river discharge and nitrification contributed 81% and 12% to the NO₃⁻ pool, respectively. A major portion (68%) of the NH₄⁺ was from river discharge, with the remainder likely from sewage and the aquitard-aquifer system. Our study demonstrates that internal nitrification can potentially be of pivotal importance in determining the NO₃⁻ level in an estuary and its export to coastal waters.

Laboratory experiments with intact sediment cores from a hypertrophic very windy exposed shallow lake were conducted to assess the combined effect of anoxia and sediment resuspension on phosphorus (P) dynamics after adding different P adsorbents (CFH-12® and Phoslock®). In this study we hypothesize that the addition of geoengineering materials will increase P retention in the sediment even at the worst physic-chemical conditions such as anoxia and sediment resuspension. Both adsorbents significantly reduced the P release from the sediments after a 54 days-anoxic incubation period (CFH-12® by 85% and Phoslock® by 98%) and even after resuspension events (CFH-12® by 84% and Phoslock® by 88%), indicating that both adsorbents are suitable P inactivating agents for restoring shallow eutrophicated lakes under such circumstances. CFH-12® did not release dissolved Fe to the water column neither after the anoxic period nor after resuspension events compared to Control (no adsorbents addition). The La concentration was significantly higher in Phoslock® (3.5–5.7 μg L⁻¹) than in Control at all sampling days but it was not affected by resuspension. The high efficiency in P removal under anoxia and resuspension, the low risk of toxicity and the high maximum adsorption capacity makes CFH-12® a promising adsorbent for lake restoration. Nevertheless, further research about the influence of other factors (i.e. pH, alkalinity, interfering substances or strict anoxia) on the performance of CFH-12® is needed.

Pyraclostrobin is a widely used and highly efficient fungicide that also has high toxicity to aquatic organisms, especially fish. Although some research has reported the toxic effects of pyraclostrobin on fish, the main toxic pathways of pyraclostrobin in fish remain unclear. The present study has integrated histopathological, biochemical and hematological techniques to reveal the main toxic pathways and mechanisms of pyraclostrobin under different exposure routes. Our results indicated that pyraclostrobin entered fish mainly through the gills. The highest accumulation of pyraclostrobin was observed in the gills and heart compared with accumulation in other tissues and gill tissue showed the most severe damage. Hypoxia symptoms (water jacking, tummy turning and cartwheel formation) in fish were observed throughout the experiment. Taken together, our results suggested that the gills are important target organs. The high pyraclostrobin toxicity to gills might be associated with oxidative damage to the gills, inducing alterations in ventilation frequency, oxygen-carrying substances in blood and disorders of energy metabolism. Our research facilitates a better understanding of the toxic mechanisms of pyraclostrobin in fish, which can promote the ecotoxicological research of agrochemicals on aquatic organisms.

1,2-dibromo-4-(1,2-dibromoethyl)-cyclohexane (DBE-DBCH) is a brominated flame retardant used in commercial and industrial applications. The use of DBE-DBCH containing products has resulted in an increased release into the environment. However, limited information is available on the long-term effects of DBE-DBCH and its effects in aquatic invertebrates. Thus, the present study was aimed at determining how DBE-DBCH diastereomers (αβ and γδ) affects aquatic invertebrates using Daphnia magna as a model organism. Survival, reproduction, feeding, swimming behavior and toxicogenomic responses to environmental relevant concentrations of DBE-DBCH were analyzed. Chronic exposure to DBE-DBCH resulted in decreased lifespan, and reduced fecundity. Expression of genes involved in reproductive processes, vtg1 and jhe, were also inhibited. DBE-DBCH also induced hypoxia by inhibiting the transcription of genes involved in heme biosynthesis and oxygen transport. Furthermore, DBE-DBCH also inhibited feeding resulting in emptiness of the alimentary canal. Increased expression of the stress response biomarkers was observed following DBE-DBCH exposure. In addition, DBE-DBCH diastereomers also altered the swimming behavior of Daphnia magna. The present study demonstrates that DBE-DBCH cause multiple deleterious effects on Daphnia magna, including effects on reproduction and hormonal systems. These endocrine disrupting effects are in agreement with effects observed on vertebrates. Furthermore, as is the case in vertebrates, DBE-DBCH γδ exerted stronger effects than DBE-DBCH αβ on Daphnia magna. This indicate that DBE-DBCH γδ has properties making it more toxic to all so far studied animals than DBE-DBCH αβ.

Hypoxia is a major stressor in aquatic environments and it is frequently linked with excess nutrients resulting from sewage effluent discharges and agricultural runoff, which often also contain complex mixtures of chemicals. Despite this, interactions between hypoxia and chemical toxicity are poorly understood. We exposed male three-spined stickleback during the onset of sexual maturation to a model anti-androgen (flutamide; 250 μg/L) and a pesticide with anti-androgenic activity (linuron; 250 μg/L), under either 97% or 56% air saturation (AS). We assessed the effects of each chemical, alone and in combination with reduced oxygen concentration, by measuring the transcription of spiggin in the kidney, as a marker of androgen signalling, and 11 genes in the liver involved in some of the molecular pathways hypothesised to be affected by the exposures. Spiggin transcription was strongly inhibited by flutamide under both AS conditions. In contrast, for linuron, a strong inhibition of spiggin was observed under 97% AS, but this effect was supressed under reduced air saturation, likely due to interactions between the hypoxia inducible factor and the aryl hydrocarbon receptor (AhR) pathways. In the liver, hypoxia inducible factor 1α was induced following exposure to both flutamide and linuron, however this was independent of the level of air saturation. This work illustrates the potential for interactions between hypoxia and pollutants with endocrine or AhR agonist activity to occur, with implications for risk assessment and management.

The Yangtze River, which is the largest in Euro-Asian, receives tremendous anthropogenic nitrogen input and is typically characterized by severe eutrophication and hypoxia. Two major processes, denitrification and anaerobic ammonium oxidation (anammox), play vital roles for removing nitrogen global in nitrogen cycling. In the current study, sediment samples were collected from both latitudinal and longitudinal transects along the coastal Yangtze River and the East China Sea (ECS). We investigated community composition and distributions of nosZ gene-encoded denitrifiers by high throughput sequencing, and also quantified the relative abundances of both denitrifying and anammox bacteria by q-PCR analysis. Denitrifying communities showed distinct spatial distribution patterns that were impacted by physical (water current and river runoffs) and chemical (nutrient availability and organic content) processes. Both denitrifying and anammox bacteria contributed to the nitrogen removal in Yangtze Estuary and the adjacent ECS, and these two processes shifted from coastal to open ocean with reverse trends: the abundance of nosZ gene decreased from coastal to open ocean while anammox exhibited an increasing trend based on quantifications of hzsB and 16S rRNA genes. Further correspondence correlation analysis revealed that salinity and nutrients were the main factors in structuring composition and distribution of denitrifying and anammox bacteria. This study improved our understanding of dynamic processes in nitrogen removal from estuarine to open ocean. We hypothesize that denitrification is the major nitrogen removal pathway in estuaries, but in open oceans, low nutrient and organic matter concentrations restrict denitrification, thus increasing the importance of anammox as a nitrogen removal process.

Hydrocarbons contamination and hypoxia are two stressors that can coexist in coastal ecosystems. At present, few studies evaluated the combined impact of these stressors on fish physiology and behavior. Here, we tested the effect of the combination of hypoxia and petrogenic hydrocarbons on the anti-predator locomotor performance of fish. Specifically, two groups of European sea bass (Dicentrarchus labrax) were exposed to clean water (Ctrl) or oil-contaminated water (Oil). Subsequently, fish of both groups were placed in normoxic (norx) or hypoxic (hyp) experimental tanks (i.e. four groups of fish were formed: Ctrl norx, Ctrl hyp, Oil norx, Oil hyp). In these tanks, escape response was elicited by a mechano-acoustic stimulus and recorded with a high speed camera. Several variables were analyzed: escape response duration, responsiveness (percentage of fish responding to the stimulation), latency (time taken by the fish to initiate a response), directionality (defined as away or toward the stimulus), distance-time variables (such as speed and acceleration), maneuverability variables (such as turning rate), escape trajectory (angle of flight) and distancing of the fish from the stimulus. Results revealed (i) effects of stressors (Ctrl hyp, Oil norx and Oil hyp) on the directionality; (ii) effects of Oil norx and Oil hyp on maneuverability and (iii) effects of Oil hyp on distancing. These results suggest that individual stressors could alter the escape response of fish and that their combination could strengthen these effects. Such an impact could decrease the probability of prey escape success. By investigating the effects of hydrocarbons (and the interaction with hypoxia) on the anti-predator behavior of fish, this work increases our understanding of the biological impact of oil spill. Additionally, the results of this study are of interest for oil spill impact evaluation and also for developing new ecotoxicological tools of ecological significance.