One-year monitoring of core biomarker and digestive enzyme responses in transplanted zebra mussels (Dreissena polymorpha).

A 12-month active biomonitoring study was performed in 2008-2009 on the Vesle river basin (Champagne-Ardenne, France) using the freshwater mussel Dreissena polymorpha as a sentinel species; allochthonous mussels originating from a reference site (Commercy) were exposed at four sites (Bouy, Sept-Saulx, Fismes, Ardre) within the Vesle river basin. Selected core biomarkers (acetylcholinesterase (AChE) activity, glutathione-S transferase (GST) activity, metallothionein concentration), along with digestive enzyme activities (amylase, endocellulase) and energy reserve concentrations (glycogen, lipids), were monitored throughout the study in exposed mussels. At the Fismes and Ardre sites (downstream basin), metallic and organic contamination levels were low but still high enough to elicit AChE and GST activity induction in exposed mussels (chemical stress); besides, chemical pollutants had no apparent deleterious effects on mussel condition. At the Bouy and Sept-Saulx sites (upstream basin), mussels obviously suffered from adverse food conditions which seriously impaired individual physiological state and survival (nutritional stress); food scarcity had however no apparent effects on core biomarker responses. Digestive enzyme activities responded to both chemical and nutritional stresses, the increase in energy outputs (general adaptation syndrome-downstream sites) or the decrease in energy inputs (food scarcity-upstream sites) leading to mid- or long-term induction of digestive carbohydrase activities in exposed mussels (energy optimizing strategy). Complex regulation patterns of these activities require nevertheless the use of a multi-marker approach to allow data interpretation. Besides, their sensitivity to natural confounding environmental factors remains to be precised.

Polychlorinated biphenyls in freshwater salmonids from the Kerguelen Islands in the Southern Ocean.

The Subantarctic Kerguelen Islands (49°S, 70°E) contain freshwater ecosystems among the most isolated in the world. Concentrations of polychlorinated biphenyls (PCBs) were assessed in the muscle of 48 brook trout and 38 brown trout caught during summer and spring 2006 in the rivers, lakes and ponds of Kerguelen. The sum of 29 PCBs averaged 404 and 358 ng g(-1) lipid, and dioxin-like PCB was 19 and 69 ng g(-1) lipid, in brook and brown trout, respectively. The values showed a high variability and some fish accumulated PCBs at levels similar to those of fish from impacted areas. While inter-sex differences were limited, the season and the morphotype appeared to have the most influence. Fish captured in summer had muscle PCB concentrations about three times higher than those caught in spring and the 'river' morphotype of brook trout showed the highest PCB levels.

One-year monitoring of reproductive and energy reserve cycles in transplanted zebra mussels (Dreissena polymorpha).

A 12-month active biomonitoring study was performed in 2008-2009 on a northern French river system using the freshwater mussel Dreissena polymorpha as a sentinel species. Allochtonous mussels originating from a reference site (Commercy) were caged at four sites (Bouy, Sept-Saulx, Fismes, Ardre) within the Vesle River basin. The main objective of the study was to characterize the influence of biotic (sex, food availability) and abiotic (temperature, chemicals) factors on the reproductive and energy reserve (glycogen, lipids) cycles of exposed mussels. Both cycles were markedly disturbed at the Bouy and Sept-Saulx sites where the lowest chlorophyll a levels were recorded during the study. At these sites, mussels obviously faced a negative energy balance, as confirmed by the impairment of their physiological state and byssal attachment. At other exposure sites, reproductive and energy reserves cycles were less impacted but were still dependent on the nutritional state of mussels. The latter appeared as a significant natural confounding factor in ecotoxicological survey performed in low polluted areas.

Lessons from a transplantation of zebra mussels into a small urban river: An integrated ecotoxicological assessment.

It is often difficult to evaluate the level of contamination in small urban rivers because pollution is mainly diffuse, with low levels of numerous substances. The use of a coupled approach using both chemical and biological measurements may provide an integrated evaluation of the impact of micro-pollution on the river. Zebra mussels were transplanted along a metal and organic pollution gradient in spring 2008. For two months, mussels and water samples were collected from two sites every two weeks and analyzed for metal and PAH content as well as water physicochemical parameters. Diffusive gradients in thin film (DGT) were also used to assess levels of labile metals. Exposure of mussels to contaminants and potential impact were evaluated using physiological indices and various biomarkers including condition index (CI), defense mechanisms (glutathione-S-transferase: GST), digestive enzymes (amylase and cellulase) and genotoxicity (micronucleus test: MN and comet assay: CA). For most contaminants, the water contamination was significantly higher downstream. Bioaccumulation in zebra mussels was related to water contamination in the framework of the biodynamic model, which allowed us to take into account the biological dilution that was caused by the growth of soft tissue downstream. Thus, metal influxes were on average two times higher downstream than upstream in particular for Zn, Cr, Cu and Cd. Significant differences in condition index were observed (final CI was 0.42 ± 0.03 downstream and 0.31 ± 0.03 upstream) reflecting a better food availability downstream. Moreover a significant decrease of GST activity and digestive enzymes activity in the cristalline style was observed downstream. Interpreting this decrease requires considering not only micro-pollution but also the trophic status related to the water's physicochemistry. The MN test and the CA on gill cells highlighted genotoxicity in mussels transplanted downstream compared to upstream.

Effects of metals on feeding rate and digestive enzymes in Gammarus fossarum: an in situ experiment.

The feeding activity and afterward the assimilation of the products resulting of the food digestion, allow organisms to obtain energy useful for growth, maintenance and reproduction. These biological parameters may be studied to assess the impact of contaminants on the energy metabolism of organisms, which could induce potential effects at an individual level. The studied species was an amphipod Gammarus fossarum, which has a high ecological relevance since it is widespread in European streams and plays a major role in the breakdown of leaf litter. Thus some G. fossarum were transplanted in four sites of a river characterized by metal contamination (Amous River, France). The following parameters were studied: digestive enzymes activities (esterase, beta-glucosidase, beta-galactosidase, amylase and endoglucanase), feeding rate, metal bioaccumulation and survival. Results showed a strong relationship between digestive enzymes activities, feeding rate and metal contents.

Importance of metallothioneins in the cadmium detoxification process in Daphnia magna.

Good knowledge of the relationship between toxic metals and biological systems, particularly the sub-cellular fraction, could be a suitable early indicator of toxic effects. These effects and the sub-cellular behaviour of cadmium were studied with a widely used species in freshwater toxicity bioassays, Daphnia magna. In spite of this very commonplace usage in ecotoxicological studies, very few data are available on its toxicant metabolism and in particular metal homeostasis. Combining multi-tools analysis, a soluble protein was found: it is heat-stable, rich in sulfhydryl groups (differential pulse polarography), characterised by a molecular mass of approximately 6.5 kDa, with a G-75 chromatographic profile corresponding to the rabbit metallothioneins monomer, with few if any aromatic-containing amino acids, it binds metals (e.g. Cd, Cu), and its concentration increases with Cd exposure. This evidence led us to hypothesise that metallothioneins (MTs) are present in D. magna. Up to 75% of the Cd body burden with Cd exposure is bound to the MTs fraction. The increase in the Cd concentration in the surrounding medium and concomitantly in daphnids induces sub-cellular reorganisation of essential metals such as Cu and Zn. The rate of metals in the soluble cellular fraction and associated with MTs increases with the Cd body burden. Monitoring sub-cellular distribution of metals after exposure in the natural environment could be very useful for ecotoxicological assessment.

Metallothionein induction in aquatic oligochaete tubifex tubifex exposed to herbicide isoproturon.

Metallothioneins (MTs) are low-molecular-weight proteins mainly involved in metal ion detoxification. Recently it has been demonstrated that MTs participate in several cellular functions such as regulation of growth and antioxidative defenses. Moreover, pesticides can induce their synthesis. The aim of the current work was to determine the effects of isoproturon, either pure or formulated as Matin (suspension containing an isoproturon concentration of 500 g. L(-1)), on the metallothionein and total protein contents of the aquatic worm Tubifex tubifex. MT levels in exposed worms increased significantly after 7 and 15 days of exposure to a concentration of the herbicide of 50 mg. L(-1). Isoproturon reduced the metal (Cu, Zn, and Cd) content of metallothioneins, and it also increased the total protein content of the worms. These results suggest that MT induction may not be considered a specific biomarker of metal exposure but that it can be used as a nonspecific biomarker of the effect of isoproturon effect in aquatic worms.

Metallothioneins in liver of Rutilus rutilus exposed to Cu2+. Analysis by metal summation, SH determination and spectrofluorimetry.

Metallothioneins (MTs) have important roles in the homeostasis of essential metals and in the detoxication of heavy metals. They also represent a potential indicator of aquatic contamination by metals. Routine methods are needed for MTs quantification in ecotoxicological studies. This paper investigates the possibility to use the spectrofluorescent properties of Cu-MTs for MTs quantification. Cu displacement of metals coordinated to MTs and spectrofluorimetric determination of the obtained Cu-MTs was tested with commercial MTs and Cu2+-induced MTs in roach liver (Rutilus rutilus). Results of this original and simple spectrofluorimetric quantification of MTs presented a good correlation with data obtained with SH quantification, but not with metal summation evaluation of MTs (analysis of Zn, Cu and Cd coordinated to MTs). The three methods showed an clear induction of MTs in roach liver after 7 days of Cu2+ exposure. After 14 days of contamination, a reduction of hepatic MTs content was observable and not correlated to liver recovery. Results show that this low cost spectrofluorimetric method is useful to quantify MTs.

Biochemical and (ultra)structural hepatic perturbations of Brachydanio rerio (Teleostei, Cyprinidae) exposed to two sublethal concentrations of copper sulfate.

The aim of this study was to compare biochemical and (ultra)structural perturbations induced by Cu in the liver of Brachydanio rerio exposed for 14 days to sublethal concentrations of copper and then replaced for 14 days to clean water. Toxicity of Cu was clearly demonstrated: simultaneously to Cu accumulation, the liver developed large lysed areas and hepatocytic alterations. However, the majority of the parenchyma was composed of functional dark hepatocytes displaying typical feature of increased metabolism: development of rough reticulum, increase in size of nucleus and nucleolus, glycogenic depletion. Increase in hepatic protein content and of anti-oxidative defences (glutathione content, catalase and glutathione-S-transferase (GST) activities) indicated that the overall response of the liver was adaptative. In all hepatocytes a cord-like structure of the nucleoli was suspected to be associated to metal deposition. After 14 days depuration, the liver still contained high Cu concentrations and the hepatic alterations were not reversed. Such complementary studies are necessary for a better understanding of the deleterious effects of pollutants and for the development of biomarkers for metal toxicity.

Toxic effects and bioaccumulation of cadmium in the aquatic oligochaete Tubifex tubifex.

Although Tubifex tubifex (Oligochaeta, Tubificida) has been proposed as a test organism for ecotoxocological studies, very few data concerning sublethal toxicity and bioaccumulation are available on this worm. The aims of this work were to assess the toxicity of cadmium, one of the most toxic metals frequently encountered in polluted areas, on T. tubifex and the ability of the worm to accumulate this metal. Acute toxicity was analyzed by measurement of the 96-h LC(50) and daily survival rates. Results indicated that T. tubifex undergoes an adaptation period to Cd, the duration of which decreases with increasing Cd concentration. The various parameters affecting toxicity are discussed. Sublethal toxicity was studied by scanning electron microscopy. Observations revealed that Cd induced autotomy of the caudal region and mucus production. Autotomy is proposed as a criterion for sublethal toxicity. The results of bioaccumulation studies revealed that Cd is highly and rapidly taken up by the worm, suggesting involvement of efficient detoxification mechanisms. Consequently, the ability to accumulate large amounts of Cd may represent a potential toxicological risk to predators of the worm if Cd is accumulated in bioavailable forms.

Effects of platinum (Pt4+) on Lumbriculus variegatus Müller (Annelida, Oligochaetae): acute toxicity and bioaccumulation.

The acute toxicity and bioaccumulation (rates, kinetic) of tetravalent platinum in Lumbriculus variegatus under different physicochemical conditions (temperature and total water hardness) were investigated. Increased Pt4+ concentration (from 0.05 to 50 mg/L), exposure (up to 30 days), temperature (from 4 to 20 degrees C) and decreasing water hardness (from 300 to 0 mg/L CaCO3) increased Pt toxicity. The metal accumulated at a constant rate that was concentration, temperature and time dependent. The median lethal concentration (96h LC50) varied greatly from 0.397 mg/L in distilled water to 30 mg/L in the hard water from Champagne. Thus, L. var. can tolerate high levels of Pt. As a result, L. var. can be used in the laboratory to analyze mechanisms of adaptation to the induced stress and in the field as an indicator of Pt pollution.