Search

International audience; Variable environmental availability of metal ions represents a constant challenge for most organisms, so that during evolution, they have optimised physiological and molecular mechanisms to cope with this particular requirement. Metallothioneins (MTs) are proteins that play a major role in metal homeostasis and as a reservoir. The MT gene/protein systems of terrestrial helicid snails are an invaluable model for the study of metal-binding features and MT isoform-specific functionality of these proteins. In the present study, we characterised three paralogous MT isogenes and their expressed products in the escargot (Cantareus aspersus). The metal-dependent transcriptional activation of the three isogenes was assessed using quantitative Real Time PCR. The metal-binding capacities of the three isoforms were studied by characterising the purified native complexes. All the data were analysed in relation to the trace element status of the animals after metal feeding. Two of the three C. aspersus MT (CaMT) isoforms appeared to be metal-specific, (CaCdMT and CaCuMT, for cadmium and copper respectively). A third isoform (CaCd/CuMT) was non-specific, since it was natively recovered as a mixed Cd/Cu complex. A specific role in Cd detoxification for CaCdMT was revealed, with a 80-90% contribution to the Cd balance in snails exposed to this metal. Conclusive data were also obtained for the CaCuMT isoform, which is involved in Cu homeostasis, sharing about 30-50% of the Cu balance of C. aspersus. No apparent metal-related physiological function was found for the third isoform (CaCd/CuMT), so its contribution to the metal balance of the escargot may be, if at all, of only marginal significance, but may enclose a major interest in evolutionary studies.

To enlarge the possibilities of using organisms of the soil fauna to assess the bioaccumulative potential of chemicals, the kinetic of soil cadmium (Cd) transfer to the terrestrial gastropod Helix aspersa was investigated under laboratory conditions during a long-term experiment (6 months). During the exposure phase (3 months), juvenile snails were subjected to three different concentrations of Cd spiked in artificial ISO soil (ISO 0, 20 and 100 mg Cd kg−1) and to a field soil (ME4) industrially contaminated by 20 mg Cd kg −1. For both soils, internal steady-state Cd concentrations were reached in the viscera of the snails, the main storage organ for Cd, after 2 weeks of exposure whatever the Cd concentration in soil. The equilibrium concentrations in the viscera were 0.7 (±0.1), 11.3 (±2.4), 73.3 (±4.8) and 6.3 (±1.3) mg Cd kg−1 dry mass for ISO 0, ISO 20, ISO 100 and ME4, respectively. During the depuration phase (3 months), from 0 to 52% of the accumulated Cd in the viscera were removed by excretion or relocation in the foot. However, the snails were not able to depurate down to initial concentrations. Data were modelled by integrating a specific growth rate constant into one-compartment toxicokinetic models. This allowed the calculation of Cd uptake rates that can be used as indicators of metal bioavailability. Since this parameter was found to be lower for snails exposed to the field soil ME4, we concluded that lower Cd bioavailability in this field soil was responsible of the lower transfer to the snails compared to the ISO 20 soil, even though they were polluted to similar extents. Internal validation showed that the toxicokinetic models could be applied for predictive purposes, promising for the development of a bioaccumulation directive for terrestrial environment. Keywords: Accumulation, Depuration, Bioavailability, Modelling, Long-term exposure

International audience; Isotopic dilution techniques were initially used to evaluate the bioavailability of trace metals contained in soils to plants, i.e., the phytoavailability. Here, we use for the first time an isotopic technique to evaluate the zooavailability of cadmium (Cd), i.e., the bioavailability to an animal organism. A terrestrial invertebrate, the snail Helix aspersa, was exposed for 14 days to a polluted soil that was spiked with 109Cd. Isotopic composition of snail tissues was then determined, allowing the computing of the L value, which can be considered as a biological estimate of the bioavailable Cd pool in the soil. It showed that the bioavailable pool of Cd to H. aspersa represented 58% of the total soil Cd. The Evalue, a chemical estimate of the soil Cd labile pool, was calculated with short-term isotopic exchange kinetics experiments and a time-dependent model describing the isotopic exchange over time. The E(14 days) value indicated that the labile soil Cd pool represented 49.6% of the total soil Cd. The L value was significantly higher than the E value, showing that snails accessed the nonlabile soil Cd pool, generally considered as nonbioavailable. The nonlabile pool contributed for 16% to the total Cd accumulated by snails. These results showed that the uptake of Cd in the nonlabile pool by soil organisms could be important and should be considered in risk assessment procedures for metal polluted soils.

International audience; Juvenile Helix aspersa snails exposed in field microcosms were used to assess the transfer of Cd, Cu, Ni, Pb and Zn from forest soils amended with liquid and composted sewage sludge. Zn concentrations and contents were significantly higher in snails exposed to liquid and composted sludge after 5 and 7 weeks of exposure, when compared with control. Trends were less clear for the other metals. Present results show that Zn, among the cocktail of metallic trace elements (MTE) coming from sewage sludge disposal, represents the principal concern for food chain transfer and secondary poisoning risks. The microcosm design used in this experiment was well suited for relatively long-term (about 2 months) active biomonitoring with H. aspersa snails. The snails quickly indicated the variations of MTE concentrations in their immediate environment. Therefore, the present study provides a simple but efficient field tool to evaluate MTE bioavailability and transfer.

To evaluate the contribution of digestive and epithelial transfer of Cd from soil to snail, Helix aspersa was exposed to an artificial substrate contaminated with 0,100,500, and 1000 μg Cd · g −1 for 4 weeks under laboratory conditions. Two modes of exposure were used: (1) the snails were in direct contact with the substrate (DC) or (2) were separated from substrate with a perforated plate (no contact (NC)) which allowed ingestion of substrate but avoided epithelial contact. Cd concentrations in DC snails were twice as high as in NC snails. The bioaccumulation factors were 0.51±0.13 in DC snails and 0.26±0.04 in NC snails. Dose-dependent growth inhibition was noted in DC snails (500 EC 50 μg · g −1 and EC10 around 100 μg · g −1 ) while only slight effects were noted in NC snails. Internal Cd concentrations in DC and NC snails were linearly correlated with growth effect whatever the contamination route.

International audience; The transfer and the toxic effects of Cd were studied in an experimental food chain involving the snail Helix aspersa as prey organism and one of its natural predators, the carabid beetle Chrysocarabus splendens. Juvenile snails were fed plant-based food enriched with 0, 10, 50 and 100 microg g(-1) of Cd, then were offered as prey to beetle larvae from egg hatching to pupation stage. Cd concentrations in snail tissues increased with increasing Cd concentration in food and with duration of exposure. Bioaccumulation factors ranged from 1.87 to 3.39, showing that H. aspersa snails, even in their early life stages, belong to macroconcentrator species for Cd. No significant reduction of snail consumption by beetles was found in exposed groups. Cd concentrations in beetle larvae remained very low (lower than 1 microg g(-1) for all groups), demonstrating a very effective regulation capacity in beetle larvae. However, Cd concentrations in highest exposed groups were higher than those found in control groups. Cd contents in adult beetles were lower than in larvae, showing a loss of Cd during metamorphosis. Despite the low Cd concentrations found in beetles, their exposure to Cd contaminated snails led to 31% of mortality, which occurred only during pupation and for the highest exposure level. No clear sublethal effects were found. These results showed that snails inhabiting heavily polluted areas may represent a risk of secondary poisoning for predatory invertebrates and provided quantitative data on the transfer of Cd between two compartments of a terrestrial food chain.