In this study, exposure and ecotoxicity data of six human pharmaceuticals (carbamazepine, clofibric acid, diclofenac, ofloxacin, propranolol, and sulfamethoxazole) were collected, including our own experimental data and literature data. From this data collection, the two-tiered European draft guideline on the environmental risk assessment of human pharmaceuticals was tested. Measured environmental concentrations in effluents from France and in effluents and surface waters from Germany were compared to the predicted environmental concentrations (PECs) in both countries. In a similar manner, predicted no-effect concentrations (PNECs) derived from acute data and PNECs derived from chronic data were estimated for each pharmaceutical and corresponding PEC/PNEC ratios then were compared in both countries. Globally, results demonstrated that all environmental concentrations (predicted or measured) for each considered pharmaceutical exceeded the 10-ng/L cutoff value, which requires the implementation of the second-tier assessment based on ecotoxicity data. Moreover, the six pharmaceuticals showed a relatively limited acute toxicity, and carbamazepine and propranolol were inaccurately identified as having negligible risks under the current European draft procedure. Such results lead to discussion of the actual procedure on pharmaceuticals, especially on the need of appropriate ecotoxicity tests.

No abstract

EXECUTIVE SUMMARYOne of the most internationally used bioassays for toxicity screening of chemicals and for toxicity monitoring of effluents and contaminated waters is the acute toxicity test with daphnid crustaceans, and in particular that performed with Daphnia magna. Standard methods have been developed for this assay that were gradually endorsed by national and international organisations dealing with toxicity testing procedures, in view of its application within a regulatory framework. As for all toxicity tests, the organisms used for the acute D. magna assay have to be obtained from live stocks which are cultured in the laboratory on live food (micro-algae). Unsurprisingly the various standard protocols of this particular assay differ -at least to a certain extent -with regard to the test organism culturing conditions. In addition, some technical aspects of the toxicity test such as the effect criterion (mortality of immobility), the exposure time, the type of dilution water, etc., also vary from one standard to another. Although this particular assay is currently used in many countries, the technical and biological problems inherent in year-round culturing and availability of the biological material and the culturing/maintenance costs of live stocks restrict its application to a limited number of highly specialised laboratories. This fundamental bottleneck in toxicity testing triggered investigations which brought forward the concept of "microbiotests" or "small-scale" toxicity tests. "Culture/maintenance free" aquatic microbiotests with species of different phylogenetic groups were developed in the early 1990s at the Laboratory for Environmental Toxicology and Aquatic Ecology at the Ghent University in Belgium. These assays which were given the generic name "Toxkits", are unique in that they employ dormant stages ("cryptobiotic eggs") of the test species, which can be stored for long periods of time and "hatched" at the time of performance of the assays. One of these microbiotests is the Daphtoxkit F magna, which is currently used in many laboratories worldwide for research as well as for toxicity monitoring purposes. The microbiotest technology has several advantages in comparison to the "traditional" tests based on laboratory cultures, especially its independence of the stock culturing burden. However, the acceptance (or possible non-acceptance) of performing assays with test organisms obtained from "dormant eggs" should be clearly dictated by the "sensitivity" and "precision" criteria of the former assays in comparison to the latter. The first part of this review therefore thoroughly reviews the scientific literature and of data obtained from various laboratories for assays performed with either D. magna test organisms obtained from lab cultures or hatched from dormant eggs. Attention has focused on data of quality control tests performed on reference chemicals, and in particular on potassium dichromate (K 2 Cr 2 O 7 ) for which an acceptability range of 0.6-2.1 mg·L -1 has been set in ISO standard 6...

Fluoxetine is a serotonin re-uptake inhibitor, generally used as an antidepressant. It is suspected to provoke substantial effects in the aquatic environment. This study reports the effects of fluoxetine on the life cycle of four invertebrate species, Daphnia magna, Hyalella azteca and the snail Potamopyrgus antipodarum exposed to fluoxetine spiked-water and the midge Chironomus riparius exposed to fluoxetine-spiked sediments. For D. magna, a multi-generational study was performed with exposition of newborns from exposed organisms. Effects of fluoxetine could be found at low measured concentrations (around 10microgl(-1)), especially for parthenogenetic reproduction of D. magna and P. antipodarum. For daphnids, newborns length was impacted by fluoxetine and the second generation of exposed individuals showed much more pronounced effects than the first one, with a NOEC of 8.9microgl(-1). For P. antipodarum, significant decrease of reproduction was found for concentrations around 10microgl(-1). In contrast, we found no effect on the reproduction of H. azteca but a significant effect on growth, which resulted in a NOEC of 33microgl(-1), expressed in nominal concentration. No effect on C. riparius could be found for measured concentrations up to 59.5mgkg(-1). General mechanistic energy-based models showed poor relevance for data analysis, which suggests that fluoxetine targets specific mechanisms of reproduction.

The method concerning in vivo proton HR-MAS NMR metabolic profiling of the freshwater cladoceran Daphnia magna is presented. Viability tests of D. magna under different spinning rates were performed. All surviving daphnids after analysis have developed eggs and embryos like control animals. Better survival rate at the slowest rotation speed were observed. The maximum length of analysis during which the integrity of the daphnid is maintained was assessed. The recorded proton spectra of in vivo daphnia were attributed to lipids from the triglycerol category. Saturated and unsaturated omega-3 like fatty acid moieties of triacylglycerol were well identified. The relationship between physiological state of daphnids and lipid profile are discussed.

No abstract

Regulatory assessments of metal toxicity on freshwater organisms assume that toxic effects are caused by dissolved metals. In aquatic systems, organisms are exposed to both dissolved and particulate-bound metals. In this study, the chronic toxicity of dietary cadmium (Cd) on the reproduction and Cd body burden of Daphnia magna was investigated. Daphnids (<24 h) were successively exposed to dissolved Cd (8 h) and then to uncontaminated or contaminated algae (16 h) for 21 d. The results show a higher Cd burden in daphnids because of the addition of contaminated food and reveal that Cd uptake by D. magna from water and food was additive for the lowest Cd concentrations tested. Similar Cd distributions (cytosolic and insoluble fractions) were observed in the two groups of organisms, showing similar potential toxicity of Cd accumulated from the two exposure routes. Dietary Cd induces deleterious effects on D. magna reproduction. On the basis of Cd body burden of daphnids, the results support the claim that waterborne and dietary Cd exposures were additive in causing toxicity for Cd concentrations lower than 25 microg/L. At the highest Cd concentrations, the importance of dietary Cd on the daphnid contamination level decreases and confounding factors such as feeding rate reduction seem to appear, which induce an effect on neonate reproduction. In this study, we illustrate the need to take the dietary pathway into account in regulatory assessments and to establish effective concentrations with particulate-bound metals.

The conventional analysis of bioassays does not account for biological significance. However, mathematical models do exist that are realistic from a biological point of view and describe toxicokinetics and effects on test organisms of chemical compounds. Here we studied a biology-based model (DEBtox) that provides an estimate of a no-effect concentration, and we demonstrated the ability of such a model to adapt to different situations. We showed that the basic model can be extended to deal with problems usually faced during bioassays like time-varying concentrations or unsuitable choices of initial concentrations. To reach this goal, we report experimental data from Daphnia magna exposed to zinc. These data also showed the potential benefit of the model in understanding the influence of food on toxicity. We finally make some recommendations about the choice of initial concentrations, and we propose a test with a depuration period to check the relevance and the predictive capacity of the DEBtox model. In our experiments, the model performed well and proved its usefulness as a tool in risk assessment.