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The warming and acidification of surface waters as predicted by the IPCC leads aquatic species to face major multifaceted changes in their environment. Although teleosts have efficient regulatory systems to cope with these changes, such changes clearly have the potential to impact their physiological functions. Hence, it is crucial to estimate the ability of teleost fishes to cope with multi-stresses to predict how they will deal with future environments. In this context, we investigated the joint effect of warming and acidification on three-spined stickleback (Gasterosteus aculeatus) from the juvenile stage to adulthood, focusing on parameters linked to growth, sexual maturation, and reproduction. Juvenile sticklebacks were split in 2 climate scenarios: a "Current" scenario corresponding to the current seasonal physico-chemical parameters of the water of the "Rade de Brest" in France, and a "RCP8.5" scenario with a warming of 3 °C and an acidification of 0.4 pH units. After 7 months, fish in the RCP8.5 scenario reached the same size and mass as those in the Current scenario, but they needed greater amounts of food to reach satiety. Furthermore, the mortality rate over the experiment was higher in the RCP8.5 scenario. Muscle lipid content, an indicator of energy reserves, was lower in females in the RCP8.5 scenario, suggesting an increased need for energy to maintain homeostasis and other physiological functions or a divergence in energy allocation strategy. Moreover, females exhibited lower sexual maturation and egg quality under the RCP8.5 scenario, which could have contributed to the lower fertilisation rate observed. Males were more resilient to the RCP8.5 scenario, exhibiting only a trend for lower kidney somatic index scores. Altogether, these results suggest a delay and/or an inhibition of gametogenesis and maturation in fish in warmed and acidified waters. The analysis of blood sex steroid concentrations, brain gene expression profiles, and physiological indexes did not allow us to discriminate between a delay and an inhibition of maturation in the RCP8.5 scenario. Overall, these findings clearly indicate that there is a long-term global impact of combined acidification and warming on the mortality and reproductive performance of three-spined stickleback. [ABSTRACT FROM AUTHOR]

Water is impacted by a variety of increasing pressures, such as contaminants, including genotoxic pollutants. The proposed multi-biomarker approach at a sub-individual level gives a complementary indicator to the chemical and ecological parameters of the Water Framework Directive (WFD, 2000/60/EC). By integrating biomarkers of genotoxicity and erythrocyte necrosis in the sentinel fish species the three-spined stickleback (Gasterosteus aculeatus) through active biomonitoring of six stations of the Artois-Picardie watershed, north France, our work aimed to improve the already existing biomarker approach. Even if fish in all stations had high levels of DNA strand breaks, the multivariate analysis (PCA), followed by hierarchical agglomerative clustering (HAC), improved discrimination among stations by detecting an increase of nuclear DNA content variation (Etaing, St Rémy du Nord, Artres and Biache-St-Vaast) and erythrocyte necrosis (Etaing, St Rémy du Nord). The present work highlighted that the integration of these biomarkers of genotoxicity in a multi-biomarker approach is appropriate to expand physiological parameters which allow the targeting of new potential effects of contaminants. [ABSTRACT FROM AUTHOR]

A grass soda technical lignin (PB1000) underwent a process combining solvent fractionation and treatment with an ionic liquid (IL), and a comprehensive investigation of the structural modifications was performed by using high‐performance size‐exclusion chromatography, 31P NMR spectroscopy, thioacidolysis, and GC–MS. Three fractions with distinct reactivity were recovered from successive ethyl acetate (EA), butanone, and methanol extractions. In parallel, a fraction deprived of EA extractives was obtained. The samples were treated with methyl imidazolium bromide ([HMIM]Br) by using either conventional heating or microwave irradiation. The treatment allowed us to solubilize 28 % of the EA‐insoluble fraction and yielded additional free phenols in all the fractions, as a consequence of depolymerization and demethylation. The gain of the combined process in terms of antioxidant properties was demonstrated through 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH.) radical‐scavenging tests. Integrating further IL safety‐related data and environmental considerations, this study paves the way for the sustainable production of phenolic oligomers competing with commercial antioxidants. [ABSTRACT FROM AUTHOR]

Caging is an active biomonitoring strategy that employs a sentinel species, sometimes a species naturally absent from the studied site, in the surveillance of water bodies to verify whether biota may be at risk. The main advantage of caging is the possibility to standardize several biotic and abiotic parameters. However, little knowledge is available about the effects of confinement on physiology and metabolism of caged organisms. The aim of this study is to characterize confinement and food access restriction effects, induced via caging experiments using a multi-biomarker approach (biometric data, immunity, antioxidant, metabolic detoxication, and digestive enzymes). The study has been undertaken using the same experiment conducted in ecosystem conditions using three-spined stickleback (Gasterosteus aculeatus) during two different periods: one in April, corresponding to breeding season, and the other in October, outside breeding season. Fifteen fish were maintained for 21 days in different conditions (caged or uncaged and with or without food supply). The main result was that confinement stress had little impact on the biological markers of sticklebacks. However, the stressors seemed to increase the negative effects of food restriction on these biomarkers, when sticklebacks needed more energy, that is, during their breeding period. Outside breeding period, most investigated biomarkers were not impacted by caging. This study showed a way to specify the conditions of application and interpretation of biomarkers during active monitoring to ensure an effective, reliable diagnosis of water body quality. [ABSTRACT FROM AUTHOR]

With a growing interest in the concept of a circular economy, the use of lignocellulosic residues, such as lignins and humins, as potential renewable feedstock for biorefining processes looks increasingly promising. Many challenges remain for the sustainable use of humins, starting from the need to provide reference data reflecting actual usable feedstocks of such materials. This paper offers the first study, from this perspective, of the potential environmental fate of those materials and components, all related to furanics, a family of compounds the toxicity of which is still a matter of debate. Conventional ecotoxicity and biodegradability tests required by European regulation on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH) for the primary evaluation of environmental hazards were conducted in combination with fish immunomarker tests to study possible long‐term effects on aquatic ecosystems. These first results are promising as humins did not give rise to any immediate ecotoxicological concerns, and hence their use could be considered in environment‐friendly applications. © 2018 Society of Chemical Industry and John Wiley & Sons, Ltd [ABSTRACT FROM AUTHOR]

Determining digestive enzyme activity is of potential interest to obtain and understand valuable information about fish digestive physiology, since digestion is an elementary process of fish metabolism. We described for the first time (i) three digestive enzymes: amylase, trypsin and intestinal alkaline phosphatase (IAP), and (ii) three gut morphometric parameters: relative gut length (RGL), relative gut mass (RGM) and Zihler’s index (ZI) in threespine stickleback (Gasterosteus aculeatus), and we studied the effect of temperature and body size on these parameters. When mimicking seasonal variation in temperature, body size had no effect on digestive enzyme activity. The highest levels of amylase and trypsin activity were observed at 18°C, while the highest IAP activity was recorded at 20°C. When sticklebacks were exposed to three constant temperatures (16, 18 and 21°C), a temporal effect correlated to fish growth was observed with inverse evolution patterns between amylase activity and the activities of trypsin and IAP. Temperature (in both experiments) had no effect on morphometric parameters. However, a temporal variation was recorded for both RGM (in the second experiment) and ZI (in both experiments), and the later was correlated to fish body mass. [ABSTRACT FROM AUTHOR]

The aims of this work are to develop suitable analytical methods to determine the widely used anticonvulsant carbamazepine and 12 of its degradation/transformation products in water, sediment, fish ( Gasterosteus aculeatus) and mollusc ( Dreissena polymorpha). Protocols based on solid phase extraction for water, pressurized-liquid extraction for sediments and QuEChERS (quick easy cheap efficient rugged and safe) extraction for both organisms followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) are developed, validated and finally applied to samples collected during a 6-month experiment in outdoor mesocosms. Very low detection limits are reached, allowing environmentally realistic doses (namely, 0.05, 0.5 and 5 μg/L nominal concentrations) to be employed. The results indicate several metabolites and/or transformation products in each compartment investigated, with concentrations sometimes being greater than that of the parent carbamazepine. Biotic degradation of carbamazepine is demonstrated in water, leading to 10,11-dihydrocarbamazepine and 10,11-epoxycarbamazepine. In sediment, the degradation results in the formation of acridine, and 2- and 3-hydroxycarbamazepine. Finally, in both organisms, a moderate bioaccumulation is observed together with a metabolization leading to 10,11-epoxycarbamazepine in fish and 2-hydroxycarbamazepine in mollusc. Acridone is also present in fish. This study provides new and interesting data, helping to elucidate how chronic exposure to carbamazepine at relevant concentrations may affect impact freshwater ecosystems. [ABSTRACT FROM AUTHOR]

To get closer to the environmental reality, ecotoxicological studies should no longer consider the evaluation of a single pollutant, but rather combination of stress and their interaction. The aim of this study was to determine if responses of a fish to a sudden biological stress could be modified by a prior exposure to a chemical stress (a polymetallic contamination). For this purpose, in situ experiment was conducted in three ponds in the Haute-Vienne department (France). One pond was chosen for its high uranium concentration due to uranium mine tailings, and the two other ponds, which were not submitted to these tailings. Three-spined sticklebacks ( Gasterosteus aculeatus) were caged in these ponds for 14 days. After this period, fish were submitted to a biological stress, exerted by lipopolysaccharides injection after anesthesia, and were sacrificed 4 days after these injections for multi-biomarkers analyses (leucocyte viability, phagocytic capacity and reactive oxygen species production, antioxidant peptide and enzymes, lipid peroxidation and DNA damage). The pond which received uranium mine tailings had higher metallic concentrations. Without biological stress, sticklebacks caged in this pond presented an oxidative stress, with increasing of reactive oxygen species levels, modification of some parts of the antioxidant system, and lipid peroxidation. Caging in the two most metal-contaminated ponds resulted in an increase of susceptibility of sticklebacks to the biological stress, preventing their phagocytic responses to lipopolysaccharides and modifying their glutathione contents and glutathione-S-transferase activity. [ABSTRACT FROM AUTHOR]

This work aimed at assessing the influence of different exposure systems to perform the commonly used OECD 201 freshwater algal growth inhibition test in the context of nanoparticles hazard assessment. Two distinct TiO2nanoparticles were considered and three different exposure systems were investigated: Erlenmeyers flasks and 24-well microplates (both using an orbital shake system), and an alternative system using cylindrical vials and magnetic stirring. All three systems are in accordance with the OECD 201 test guideline recommendations. We concluded that the exposure systems applied to achieve the test can substantially affect the ecotoxicological results and the subsequent calculated ECx. The selected systems influenced both the interaction between algal cells and TiO2nanoparticles as well as the growth inhibition recorded. Disparities in ecotoxicity relative to the TiO2nanoparticles tested were also observed and are finally discussed. [ABSTRACT FROM AUTHOR]

Human activities have led to increased levels of various pollutants including metals in aquatic ecosystems. Increase of metallic concentrations in aquatic environments represents a potential risk to exposed organisms, including fish. The aim of this study was to characterize the environmental risk to fish health linked to a polymetallic contamination from former uranium mines in France. This contamination is characterized by metals naturally present in the areas (manganese and iron), uranium, and metals (aluminum and barium) added to precipitate uranium and its decay products. Effects from mine releases in two contaminated ponds (Pontabrier for Haute-Vienne Department and Saint-Pierre for Cantal Department) were compared to those assessed at four other ponds outside the influence of mine tailings (two reference ponds/department). In this way, 360 adult three-spined sticklebacks ( Gasterosteus aculeatus) were caged for 28 days in these six ponds before biomarker analyses (immune system, antioxidant system, biometry, histology, DNA integrity, etc.). Ponds receiving uranium mine tailings presented higher concentrations of uranium, manganese and aluminum, especially for the Haute-Vienne Department. This uranium contamination could explain the higher bioaccumulation of this metal in fish caged in Pontabrier and Saint-Pierre Ponds. In the same way, many fish biomarkers (antioxidant and immune systems, acetylcholinesterase activity and biometric parameters) were impacted by this environmental exposure to mine tailings. This study shows the interest of caging and the use of a multi-biomarker approach in the study of a complex metallic contamination. [ABSTRACT FROM AUTHOR]

Pharmaceuticals are emerging organic contaminants ubiquitously present in the environment due to incessant input into the aquatic compartment mainly resulting from incomplete removal in wastewater treatment plants. One of the major preoccupations concerning pharmaceuticals released into surface waters is their potential for bioaccumulation in biota, possibly leading to deleterious effects on ecosystems especially as they could affect a broad variety of organisms living in or depending on the aquatic environment. Thus, the development of accurate and sensitive methods is necessary to detect these compounds in aquatic ecosystems. Considering this need, this study deals with the analytical development of a methodology to quantify traces of diclofenac together with some of its biotic and abiotic transformation products in whole-body tissue of three-spined stickleback. A simple and reliable extraction method based on a modified QuEChERS extraction is implemented on 200 mg of fish. The detection and quantification of the ten target compounds are performed using liquid chromatography-tandem mass spectrometry. The whole process was successfully validated regarding linearity, recovery, repeatability, and reproducibility. The method limits of detection and quantification do not exceed 1 ng/g. To reproduce environmental conditions, we measured the concentration of DCF and its transformation products in three-spined sticklebacks after a 6-month exposure in mesocosms at several levels of DCF ranging from 0.05 to 4.1 μg/L. The phase I metabolite 4′-hydroxydiclofenac was detected in fish samples exposed at the highest DCF concentration. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

To characterize environmental risks linked to former uranium mines in the Limousin region of France, a study was conducted on fish health effects from uranium releases. Two private ponds were compared in this study, one with uranium contamination and one background site, upstream of the mining zone. Roach, Rutilus rutilus, were caged for 28 days in both ponds. Physico-chemical parameters of water and sediments and bioaccumulation of metals in several organs were determined. After 14 and 28 days of caging, immune, oxidative stress, biotransformation, neurotoxicity and physiological parameters were measured. Iron and aluminium were quantified in the water of both sites; however, barium and manganese were only present in the water of the uranium contaminated site. Uranium was present in both sites but at very different concentrations. The sediments from the uranium contaminated site contained high levels of radioactive elements coming from the disintegration chain of uranium. Results of biological parameters indicated stimulation of immune parameters and of oxidative stress and a decrease of AChE in fish caged in the uranium contaminated pond compared to the uranium-free pond. Overall, the results determined roach health status in the context of pollution from poly-metallic mining. The data strengthen our knowledge of the environmental risk assessment associated with radioactive substances in the environment. [ABSTRACT FROM AUTHOR]

In this study, we investigated the effects of depleted uranium (DU), the byproduct of nuclear enrichment of uranium, on several parameters related to defence system in the zebrafish, Danio rerio, using flow cytometry. Several immune cellular parameters were followed on kidney leucocytes: cell proportion, cell mortality, phagocytosis activity and associated oxidative burst and lysosomal membrane integrity (LMI). Effects of DU were tested ex vivo after 17 h of contact between DU and freshly isolated leucocytes from 0 to 500 µg DU/L. Moreover, adult zebrafish were exposed in vivo during 3 days at 20 and 250 µg DU/L. Oxidative burst results showed that DU increased reactive oxygen species (ROS) basal level and therefore reduced ROS stimulation index in both ex vivo and in vivo experiments. ROS PMA-stimulated level was also increased at 250 µg DU/L in vivo only. Furthermore, a decrease of LMI was detected after in vivo experiments. Cell mortality was also decreased at 20 µg DU/L in ex vivo experiment. However, phagocytosis activity was not modified in both ex vivo and in vivo experiments. A reduction of immune-related parameters was demonstrated in zebrafish exposed to DU. DU could therefore decrease the ability of fish to stimulate its own immune system which could, in turn, enhance the susceptibility of fish to infection. These results encourage the development and the use of innate immune analysis by flow cytometry in order to understand the effects of DU and more generally radionuclides on fish immune system and response to infectious diseases. [ABSTRACT FROM AUTHOR]

The risk posed by complex chemical mixtures in the environment to wildlife and humans is increasingly debated, but has been rarely tested under environmentally relevant scenarios. To address this issue, two mixtures of 14 or 19 substances of concern (pesticides, pharmaceuticals, heavy metals, polyaromatic hydrocarbons, a surfactant, and a plasticizer), each present at its safety limit concentration imposed by the European legislation, were prepared and tested for their toxic effects. The effects of the mixtures were assessed in 35 bioassays, based on 11 organisms representing different trophic levels. A consortium of 16 laboratories was involved in performing the bioassays. The mixtures elicited quantifiable toxic effects on some of the test systems employed, including i) changes in marine microbial composition, ii) microalgae toxicity, iii) immobilization in the crustacean Daphnia magna, iv) fish embryo toxicity, v) impaired frog embryo development, and vi) increased expression on oxidative stress-linked reporter genes. Estrogenic activity close to regulatory safety limit concentrations was uncovered by receptor-binding assays. The results highlight the need of precautionary actions on the assessment of chemical mixtures even in cases where individual toxicants are present at seemingly harmless concentrations. [ABSTRACT FROM PUBLISHER]

This work focuses on the effects of two commercial formulations of dispersants on juvenile turbot after 48 h of contamination and 15 days of recovery. Oxidative stress, gill, and immune functions were assessed in seven conditions: exposition to the water-soluble fraction of an oil, mechanical dispersion, two dispersants alone, two types of chemical dispersion and a control group. In the contaminated groups, nominal concentrations of oil and dispersants were 66 and 3.3 mg L, respectively. Dispersants alone had weak effects; the soluble fraction induced leucopenia and gill alteration. Chemical and mechanical dispersion induced similar effects. After contamination, a principal component analysis showed two distinct areas: the first one included the control and dispersants groups, the second one dispersion of the oil. After the 15-day recovery period, it was not possible to differentiate the groups. This study shows that, in the experimental conditions tested, the dispersion, either chemical or mechanical, enhances the consequences of exposure to crude oil without long-lasting consequences. [ABSTRACT FROM AUTHOR]

The aim of this study was to identify a signal that could be used as an androgen exposure indicator in the European bullhead ( Cottus sp.). For this purpose, the ultra-structure of the kidney was characterized to identify normal structure of this organ, and histological changes previously described in the kidney of breeding male bullheads were quantified using the kidney epithelium height (KEH) assay previously developed and validated for the stickleback. In the next step, the effect of trenbolone acetate (TbA), a model androgen, was assessed to identify potential androgenic regulation of bullhead kidney hypertrophy. Measurement of KEH performed on adult non-breeding male and female bullheads exposed for 14 and 21 days to 0, 1.26 and 6.50 μg/L showed that kidney hypertrophy is induced in a dose-dependent manner, confirming the hypothesis that the European bullhead possesses a potential biomarker of androgen exposure. Combined with the wide distribution of the European bullhead in European countries and the potential of this fish species for environmental toxicology studies in field and laboratory conditions, the hypothesis of a potential biomarker of androgen exposure offers interesting perspectives for the use of the bullhead as a relevant sentinel fish species in monitoring studies. Inducibility was observed with high exposure concentrations of TbA. Further studies are needed to identify molecular signals that could be more sensitive than KEH. [ABSTRACT FROM AUTHOR]

The neutral red retention assay has been proposed to determine the lysosomal membrane stability in immune cells. Nevertheless, this assay implies many examinations under a microscope at short time intervals and therefore the analysis of few samples. The present study proposes two more rapid, efficient, and sensitive sample analyses using flow cytometry method. Lysosomal presence and lysosomal membrane integrity (LMI) were evaluated on the three-spined stickleback, Gasterosteus aculeatus (L.), a well-described model fish species for aquatic ecotoxicology studies. After development of the two biomarkers, they were validated by ex vivo contamination with endosulfan and copper and by in situ sampling. These immunomarkers were clearly modulated by pollutants and their variations seemed to be correlated with leucocyte mortality. Thus, from a practical point of view, lysosomal presence and LMI may provide novel and efficient means of evaluating immune capacities and indicating the toxic effects of environmental pollution. [ABSTRACT FROM AUTHOR]

Cover Feature: Enhancing the Antioxidant Activity of Technical Lignins by Combining Solvent Fractionation and Ionic-Liquid Treatment (ChemSusChem 21/2019) B The Cover Feature b shows a cascade biorefinery process for the upgrading of technical lignins and sustainable production of phenolic oligomers of interest as substitutes for commercial antioxidants. A grass soda technical lignin undergoes a process combining solvent fractionation and treatment with an ionic liquid, and a comprehensive investigation of the structural modifications is performed. [Extracted from the article]

The article presents a study which examined how depleted uranium affects the immune parameters of zebra fish using several endpoints measured by flow cytometry. Topics discussed include the ex vivo exposure of freshly isolated leucocytes and analysis of cell sub-population, cell mortality, and oxidative burst. Reactive oxygen species stimulation, lysosomal integrity, and phagocytosis are explained.