Evaluation de l'écotoxicité de nanotubes de carbone en milieu aquatique à l'aide du modèle amphibien xenopus laevis : synthèse, protocoles d'exposition, détection et dosage

L'augmentation de la production et l'utilisation de nanotubes de carbone (NTC) soulèvent des inquiétudes vis-à-vis de leur dissémination dans l'environnement. Pourtant, les connaissances relatives à leur toxicité et leur bioaccumulation potentielles chez des organismes aquatiques sont encore très limitées. Les travaux rapportés ici sont basés sur une approche prospective à l'échelle du laboratoire, à l'aide d'un modèle aquatique, la larve d'amphibien Xenopus laevis. Les toxicités aiguë, chronique et génétique ont été évaluées après exposition des larves à des suspensions de DWNT ou de MWNT préparées selon différents protocoles de dispersion. Les effets potentiels de synergie entre des DWNT et du plomb ont par ailleurs été évalués. L'ingestion d'une quantité importante de NTC par les larves perturberait leur transit intestinal et l'assimilation de nutriments. Les réponses toxiques seraient modulées à la fois par le type de NTC, leur concentration, leur état de dispersion, la nature de l'agent dispersant, mais également par la présence de plomb et sa concentration. La localisation et le dosage des NTC dans des matrices biologiques demeurent des défis considérables. D'une part, le principe de détection de NTC par dosage isotopique du carbone (13C/12C) a été exploré chez des larves exposées à des NTC synthétisés à partir d'une source carbonée enrichie en 13C. Des essais préliminaires suggèrent la bioaccumulation hétérogène de 13C-DWNT dans le foie, mais elle reste à confirmer. D'autre part, une technique originale de dosage de DWNT a été mise au point par mesure de la permittivité complexe de suspensions de larves soumises à un champ électromagnétique dans la gamme micro-ondes.The exponential rise in the production and the use of carbon nanotubes (CNTs) raises concerns about their spread in the environment. Nevertheless, few information is available about their potential ecotoxicity and bioaccumulation in aquatic organisms. Our work was based on a forward-looking approach at the laboratory scale, using an aquatic model, the amphibian larva of Xenopus laevis. The acute, chronic and genetic toxicities were assessed after the exposure of larvae to DWCNT or MWCNT suspensions prepared according to various dispersion protocols. The potential synergistic effects of raw DWCNTs and lead were also assessed. The ingestion of CNT could disrupt the intestinal transit and the assimilation of nutrients. The toxic responses seem to be modulated both by the kind of CNTs, their concentration, and their dispersion state, the nature of the dispersing agent, as well as the presence of lead and its concentration (co-exposure). The tracking and the quantitative analysis of CNTs in biological samples are still huge challenges. On the one hand, larvae were exposed to CNTs synthesized from 13C-enriched sources in order to investigate the principle of probing their presence using the 13C/12C ratio (isotopic labeling). Preliminary studies suggest an heterogeneous biomagnification of 13C-DWCNTs in the liver, which has to be confirmed by more extensive investigations. On the other hand, a new technique for the quantitative detection of CNTs was developed on larvae exposed to DWCNTs. It is based on the dielectric relaxation of the sample suspensions in a microwave electromagnetic field which allows the extrapolation of CNT concentrations from complex permittivity measurements

Factors affecting methylmercury biomagnification by a widespread aquatic invertebrate predator, the phantom midge larvae Chaoborus

MeHg biomagnification by the phantom midge Chaoborus in relation to MeHg concentrations in their prey and its migratory behavior was investigated in two Canadian Precambrian Shield lakes. Three Chaoborus species with contrasted migratory behavior were collected in a fishless and a fish-inhabited lake. All species accumulated MeHg through their ontogenic development. In the lake inhabited by fish, all instars of Chaoborus punctipennis displayed a marked migratory behavior and were unable to biomagnify MeHg, whereas in the fishless lake, Chaoborus americanus and Chaoborus trivittatus biomagnified MeHg. Reduced biomagnification capacity of C. trivittatus, the coexisting species living with C. americanus, was also ascribed to a progressive vertical segregation with age. Growth dilution, amount and type of prey items or trophic position could not explain the different patterns of biomagnification. Our findings demonstrate that the most common invertebrate predator of temperate planktonic food webs can biomagnify mercury, contrarily to previous reports

Biocompatible polymer-assisted dispersion of multi walled carbon nanotubes in water, application to the investigation of their ecotoxicity using Xenopus laevis amphibian larvae

Carbon nanotubes (CNTs) tend to readily agglomerate and settle down in water, while the adsorption of compounds present in natural aquatic media could enhance their dispersion and stabilization in the water column. We designed a new exposure protocol to compare the biological responses of Xenopus laevis larvae exposed in semi-static conditions to size-reduced agglomerates of multi-walled carbon nanotubes (MWCNTs) in suspension in the water column and/or to larger agglomerates. Suspensions were prepared using a combination of a non-covalent functionalization with a non-toxic polymer (either carboxymethylcellulose, CMC, or gum arabic, GA) and mechanical dispersion methods (mainly ultrasonication). The ingestion of agglomerates which have settled down was incriminated in the disruption of the intestinal transit and the assimilation of nutrients, leading to acute and chronic toxicities at the highest tested concentrations. Rise in mortality, decrease in the growth rate and induction of genotoxicity from low concentrations (1 mg/L in the presence of CMC) were evidenced in presence of suspended MWCNTs in the water column. The biological responses seemed to be modulated when GA, a potential antioxidant, was used. We hypothesized that MWCNTs should interfere mainly at the surface of the gills, acting as a potential respiratory toxicant and generally inducing indirect effects

Multi-walled carbon nanotubes, natural organic matter, and the benthic diatom Nitzschia palea: “A sticky story”

Different effects of multi-walled carbon nanotubes (MWCNTs) on the freshwater diatom Nitzschia palea were examined. MWCNTs used in this study (MWCNT) were dispersed either by sonication without (MWCNTsonicated) or with a realistic concentration (10mgL-1) of Natural Organic Matter (MWCNT+NOM). A pocket-size device was designed to distinguish shading effect (using MWCNT suspensions as external filters) from total exposure effect of MWCNTsonicated and MWCNT+NOM on benthic algae. This study demonstrates that cell division was strongly inhibited after a 48 h exposure to MWCNT+NOM compared to MWCNTsonicated. This device did not yield a quantifiable contribution of shading to growth inhibition of MWCNTsonicated and below 10mgL-1 of MWCNT+NOM. In all cases, neither lethal effects nor drops in photosynthetic quantum yield were observed. After a 6-d exposure, a complete growth recovery was observed for all conditions except at the highest concentration of MWCNT+NOM. Different microscopic approaches using carbohydrates markers revealed the strong affinity between MWCNT and extracellular polymeric substances (EPS) produced by N. palea. These seem to constitute a defensive mechanism against MWCNT

Évaluation de l'écotoxicité de nanotubes de carbone en milieu aquatique à l'aide du modèle amphibien xenopus laevis (synthèse, protocoles d'exposition, détection et dosage)

TOULOUSE3-BU Sciences (315552104) / SudocSudocFranceF

Quantitative detection of carbon nanotubes in biological samples by an original method based on microwave permittivity measurements

International audienceDue to their nanoscale, morphology, and chemical composition, the tracking and thequantitative analysis of carbon nanotubes (CNTs) in biological samples still represent hugechallenges. A new technique for the quantitative and accurate detection of CNTs in variousbiological samples at different scales (whole organisms to organs) was developed usingamphibian larvae exposed to double-walled CNTs (DWCNTs). This technique is based onthe dielectric relaxation of ultra-low volume suspensions under a microwave electromagneticfield. CNT concentrations were consequently extracted from complex permittivitymeasurements at 5 GHz, making possible to quantitatively assess the animal exposure toCNTs. Our results indicate a detection threshold of 0.02 lg of DWCNTs, which is the lowestachieved in the literature to dat