Among freshwater invertebrates, Gammarus fossarum is an important test organism and is currently used in ecotoxicology for acute and chronic assays; nevertheless, reproductive toxicity test methods are not yet available for these species. In the present study, the reproductive cycle in Gammarus fossarum was characterized in order to propose a reproductive toxicity test encompassing molting, follicle growth, and embryonic development that will provide a better understanding of the mode of action of chemicals disrupting these hormone-regulated processes. A detailed description of the reproductive cycle in Gammarus fossarum was obtained. As in some amphipods, molt and reproductive cycles of G. fossarum females occur concurrently, lasting 30 d at 12°C. Each molt stage is characterized by a specific marsupial embryonic development stage and the size of developing follicles visible on the ovarian membrane. Based on these results, a 21-d reproductive toxicity test is proposed for this species. This new bioassay was applied to identify the specific impact of different stressors: cadmium, methomyl, nonylphenol, and a starvation diet. Good reproducibility was obtained for different endpoints under control conditions and throughout the experiments. Preliminary robust reference values or benchmarks were proposed for these endpoints. Cadmium was found to specially inhibit secondary vitellogenesis. Nonylphenol had a specific concentration-dependent effect on embryonic development, with an increase in the percent abnormality from a concentration of 0.05 µg/L. A restricted food diet led to a significant delay in the molt cycle, which in turn induced inhibition of secondary vitellogenesis.

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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.

The bioavailability of Cd, Cu and Zn from a metal-contaminated sediment was studied using Mytilus galloprovincialis embryos and larvae exposed to sediment elutriates. The elutriate concentrations tested were always lower than the threshold at which abnormal larval development appears. The Cd, Cu and Zn contents of the sediment, of the elutriate and of the larvae, as well as the larval growth, the condition index (CI), and the induction of metallothionein (MT) in the larvae were measured. The CI was only affected after 96 h of exposure, principally at the highest elutriate concentration (25%, corresponding to 0.1435 µg Cd l -1 and 14 µg Zn l -1). Cd, Cu and Zn bioaccumulation was observed (48 and 96 h), in whole larvae as well as in the cytosolic fraction of their tissues. For the individual treatments (controls or different degrees of exposure), the lowest metal concentrations were observed in larvae after 96 h exposure, indicating biological dilution. Metallothionein induction in larvae was observed after 96 h exposure at the lowest elutriate concentrations tested, and was always strongly correlated with increasing metal contents in the cytosolic fraction. These relationships were significant for all 3 metals studied either individually or combined. The results indicate that these 3 metals are bioavailable to M. galloprovincialis larvae and that MT induction constitutes a more sensitive indicator of heavy metal pollution than embryotoxicity or larval growth experiments. KEY WORDS: Sediment elutriate · Mytilus galloprovincialis larvae · Metal bioaccumulation · MetallothioneinResale or republication not permitted without written consent of the publisher

The feeding activity and subsequent assimilation of the products resulting from food digestion allow organisms to obtain energy for growth, maintenance and reproduction. Among these biological parameters, we studied digestive enzymes (amylase, cellulase and trypsin) in Gammarus fossarum to assess the impact of contaminants on their access to energy resources. However, to enable objective assessment of a toxic effect of decreased water quality on an organisms' digestive capacity, it is necessary to establish reference values based on its natural variability as a function of changing biotic and abiotic factors. To limit the confounding influence of biotic factors, a caging approach with calibrated male organisms from the same population was used. This study applied an in situ deployment at 23 sites of the Rhone basin rivers, complemented by a laboratory experiment assessing the influence of two abiotic factors (temperature and conductivity). The results showed a small effect of conductivity on cellulase activity and a significant effect of temperature on digestive enzyme activity but only at the lowest temperature (7 °C). The experimental conditions allowed us to define an environmental reference value for digestive enzyme activities to select sites where the quality of the water impacted the digestive capacity of the organisms. In addition to the feeding rate, this study showed the relevance of digestive enzymes as biomarkers to be used as an early warning tool to reflect organisms' health and the chemical quality of aquatic ecosystems.

Abstract-Surface sediment samples from 10 coastal locations were analyzed with respect to their physical and chemical characteristics as well as their biological quality. The toxicity of the decanted fresh and freeze-dried sediments and of their elutriates was assessed by bioassays with embryos of the oysters Crassostrea gigas. The concentration inhibiting normal embryogenesis in 20% of the larvae (EC20) was used as a measure of toxicity and compared with the physical and chemical properties of the sediments and elutriates by principal components analysis. Decanted whole sediments were one to four orders of magnitude more toxic than their elutriates because the latter have lower contaminant concentrations. The biological effects of decanted whole sediment depended mostly on the levels of heavy metals and polycyclic aromatic hydrocarbons (PAHs), while in the case of the elutriates, the toxicity was mainly correlated with the concentrations of total ammonia and of dissolved organic carbon (DOC). Freeze-dried sediments and their elutriates were up to one order of magnitude more toxic than fresh sediments and elutriates, respectively, presumably because the freeze-drying procedure increases pollutant bioavailability or causes a release of unidentified organic compounds having toxic effects.