The existence of fertile hybrids of closely related model earthworm species, Eisenia andrei and E. fetida

Lumbricid earthworms Eisenia andrei (Ea) and E. fetida (Ef) are simultaneous hermaphrodites with reciprocal insemination capable of self-fertilization while the existence of hybridization of these two species was still debatable. During the present investigation fertile hybrids of Ea and Ef were detected. Virgin specimens of Ea and Ef were laboratory crossed (Ea+Ef) and their progeny was doubly identified. 1 -identified by species-specific maternally derived haploid mitochondrial DNA sequences of the COI gene being either 'a' for worms hatched from Ea ova or 'f' for worms hatched from Ef ova. 2 -identified by the diploid maternal/paternal nuclear DNA sequences of 28s rRNA gene being either 'AA' for Ea, 'FF' for Ef, or AF/FA for their hybrids derived either from the 'aA' or 'fF' ova, respectively. Among offspring of Ea+Ef pairs in F1 generation there were mainly aAA and fFF earthworms resulted from the facilitated self-fertilization and some aAF hybrids from aA ova but none fFA hybrids from fF ova. In F2 generation resulting from aAF hybrids mated with aAA a new generations of aAA and aAF hybrids were noticed, while aAF hybrids mated with fFF gave fFF and both aAF and fFA hybrids. Hybrids intercrossed together produced plenty of cocoons but no hatchlings independently whether aAF+aAF or aAF+fFA were mated. These results indicated that Ea and Ef species, easy to maintain in laboratory and commonly used as convenient models in biomedicine and ecotoxicology, may also serve in studies on molecular basis of interspecific barriers and mechanisms of introgression and speciation. Hypothetically, their asymmetrical hybridization can be modified by some external factors

Développement de biomarqueurs d'exposition des métaux sur les fonctions physiologiques de l'annélide oligochète Eisenia fetida

Les activités métallurgiques entrainent l'accumulation d'éléments traces métalliques dans les couches . superficielles des sols, où l'on peut observer des concentrations largement supérieures au fond pédogéochimique et aux normes en vigueur. La contamination des sols est une menace pour la santé publique et la présence de grandes quantités d'éléments traces métalliques peut générer un stress susceptible d'affecter les organismes exposés à des sols pollués. Les Annélides Oligochètes vivent en contact étroit avec ces sols pollués et sont parmi les organismes vivants présentant une sensibilité exacerbée aux métaux. Très peu de choses sont connues quant à l'identification et la mise en place des mécanismes de réponse à ces métaux au niveau moléculaire. En exploitant, la conservation phylogénique observée entre espèces nous avons été capables de cloner et de caractériser un ensemble de biomarqueurs de pollution potentiels à partir des cœlomocytes de l'Annélide Oligochètes Eisenia fetida. Deux approches ont été mises en place. Premièrement, une approche qualifiée de ciblée, consistant à identifier tous les effecteurs parmi des protéines fortement conservées pour lesquels une variation lors d'une exposition métallique était reportée dans la littérature. Deuxièmement, nous avons entrepris une approche qualifiée de globale consistant en la construction de banques soustractives pour identifier chez Eisenia fetida, les gènes dont l'expression est affectée lors d'une exposition à un mélange complexe de métaux, représentatif d'un site naturel fortement contaminé. Ces deux approches ont permis l'identification de 4 candidats biomarqueurs de pollution métallique.Metallurgical activities lead to accumulation of metal trace elements in the topsoils, where one typically observes concentrations largely exceeding the natural background. Contaminations of soils are a threat for public health and the presence of high amount of metals can generate an environmental stress likely to affect the organisms exposed to polluted soils. Earthworms living in close contact with polluted soils were studied in order to better understand the physiological changes, the mechanisms of acclimation and of detoxification caused by metals. By exploiting the conservation observed between species and using molecular biology techniques, we have been able to clone and transcriptionally characterise potential biomarkers form the immune ceIls of the ecotoxicologically important earthworm species Eisenia fetida. During this Ph. D. we developed two approaches. First, a targeted approach based on bibliographie work, enabled us to identify and select effectors among highly preserved proteins for which variations following metal exposure were reported in invertebrates. Second, an open strategy was undertaken by performing a Suppression Subtractive Hybridization in order to identify genes which are differentially expressed following exposure to a complex mixture of metals representative of a strongly polluted soil. These approaches allowed the description of a group of genes which expression varies following metallic exposure as weIl as the analysis of their expression profiles may be used to analyse mechanistically the immune response to a metallic stress. Moreover, expression profiles of selected genes may constitute a signature of changes due to MTE.LILLE1-Bib. Electronique (590099901) / SudocSudocFranceF

Interest and validation of early markers of environmental genotoxicity

Le présent projet propose de mettre au point des marqueurs biologiques de génotoxicité environnementale permettant une approche intégrée de la complexité des contaminations. Les préoccupations relatives aux pollutions des sols et des sédiments sont récentes. La contamination métallique des sols par exemple, due aux activités anthropiques est cependant préoccupante. Ce, en termes de risques pour la santé humaine et environnementale, du fait de la persistance, de l accumulation, et du transfert possible de ces métaux vers les nappes souterraines et vers les chaînes trophiques. Le projet de recherche porte sur la validation au laboratoire de marqueurs de génotoxicité, mis en œuvre sur plusieurs espèces représentatives en écotoxicologie terrestre, Eisenia fetida et Trifolium repens. Des travaux préliminaires, dans le cadre d un programme Ecotoxicité des sols contaminés par les éléments traces métalliques soutenu par l ANR et les FEDER, ont permis de mettre en évidence la pertinence du test des comètes avec Trifolium vis-à-vis des sols contaminées en éléments traces métalliques ou à des hydrocarbures en mélanges (lixiviats de déchet industriels). Le test des comètes (Single Cell Gel Electrophoresis Assay) permet la détection et la quantification des dommages à l ADN (cassures double et simple brin, sites alcali-labiles) au sein d une population cellulaire, induits par certains agents environnementaux. Il permet d évaluer l atteinte au génome d organismes modèles exposés. Cette atteinte de l ADN est une composante essentielle à estimer car elle contribue à l origine des phénomènes de cancérogenèse.This project proposes to develop biomarkers of environmental genotoxicity for an integrated approach to the complexity of the contamination. Concerns about pollution of soils and sediments are recent. Metal contamination of soils, for example, due to human activities, however, is worrying. This, in terms of risks to human health and environmental, because of the persistence, accumulation, and the possible transfer of these metals into groundwater and into food chains. The research project presented consists in the validation of laboratory markers of genotoxicity, implemented on several representative species in terrestrial ecotoxicology, Eisenia fetida and Trifolium repens. Preliminary work, as part of a program "Ecotoxicity of soils contaminated with trace elements" supported by the ANR and the ERDF, helped to highlight the relevance of the comet assay with Trifolium vis-à-vis Soil contaminated by trace metals or mixtures of hydrocarbons (industrial waste leachate). The comet assay (Single Cell Gel Electrophoresis Assay) allows detection and quantification of DNA damage (DNA double and single-stranded, alkali-labile sites) in a cell population, induced by certain environmental agents. It assesses the impairment of the genome of model organisms exposed. This DNA damage is an essential component to estimate because it contributes to the origin of the phenomena of carcinogenesis.LILLE1-Bib. Electronique (590099901) / SudocSudocFranceF

Caractérisation et rôle des peptides antimicrobiens dans l'immunité innée de la sangsue Theromyzon tessulatum

LILLE1-BU (590092102) / SudocSudocFranceF

Transcriptomic underpinning of toxicant-mediated physiological function alterations in three terrestrial invertebrate taxa: A review.

Diverse anthropogenic activities often lead to the accumulation of inorganic and organic residues in topsoils. Biota living in close contact with contaminated soils may experience stress at different levels of biological organisation throughout the continuum from the molecular-genetic to ecological and community levels. To date, the relationship between changes at the molecular (mRNA expression) and biochemical/physiological levels evoked by exposures to chemical compounds has been partially established in a limited number of terrestrial invertebrate species. Recently, the advent of a family of transcriptomic tools (e.g. Real-time PCR, Subtractive Suppressive Hybridization, Expressed Sequence Tag sequencing, pyro-sequencing technologies, Microarray chips), together with supporting informatic and statistical procedures, have permitted the robust analyses of global gene expression changes within an ecotoxicological context. This review focuses on how transcriptomics is enlightening our understanding of the molecular-genetic responses of three contrasting terrestrial macroinvertebrate taxa (nematodes, earthworms, and springtails) to inorganics, organics, and agrochemicals

Relationships between metal compartmentalization and biomarkers in earthworms exposed to field-contaminated soils

Partitioning tissue metal concentration into subcellular compartments relecting 'toxicologically available' pools may provide good descriptors of the toxicological efects of metals on organisms. Here we investigated the relationships between internal compartmentalization of Cd, Pb and Zn and biomarker responses in a model soil organism: the earthworm. The aim of this study was to identify metal fractions reflecting the toxic pressure in an endogeic, naturally occurring earthworm species (Aporrectodea caliginosa) exposed to realistic field-contaminated soils. After a 21 days exposure experiment to 31 field contaminated soils, Cd, Pb and Zn concentrations in earthworms and in three subcellular fractions (cytosol, debris and granules) were quantified. Different biomarkers were measured: the expression of a metallothionein gene (mt), the activity of catalase (CAT) and of glutathione-s-transferase (GST), and the protein, lipid and glycogen reserves. Biomarkers were further combined in an integrated biomarker index (IBR). The subcellular fractionation provided better predictors of biomarkers than the total internal contents hence supporting its use when assessing toxicological bioavailability of metals to earthworms. The most soluble internal pools of metals were not always the best predictors of biomarker responses. metallothionein expression responded to increasing concentrations of Cd in the insoluble fraction (debris + granules). Protein and glycogen contents were also mainly related to Cd and Pb in the insoluble fraction. On the other hand, GST activity was better explained by Pb in the cytosolic fraction. CAT activity and lipid contents variations were not related to metal subcellular distribution. The IBR was best explained by both soluble and insoluble fractions of Cd and Pb. This study further extends the scope of mt expression as a robust and specific biomarker in an ecologically representative earthworm species exposed to field-contaminated soils. The genetic lineage of the individuals, assessed by DNA barcoding with cytochrome c oxidase subunit I, did not inuence mt expression

Modelling assisted phytoremediation of soils contaminated with heavy metals \textendash Main opportunities, limitations, decision making and future prospects

ACLThe heavy metals (HMs) soils contamination is a growing concern since HMs are not biodegradable and can accumulate in all living organisms causing a threat to plants and animals, including humans. Phytoremediation is a cost-efficient technology that uses plants to remove, transform or detoxify contaminants. In recent years, phytoremediation is entering the stage of large-scale modelling via various mathematical models. Such models can be useful tools to further our understanding and predicting of the processes that influence the efficiency of phytoremediation and to precisely plan such actions on a large-scale. When dealing with extremely complicated and challenging variables like the interactions between the climate, soil and plants, modelling before starting an operation can significantly reduce the time and cost of such process by granting us an accurate prediction of possible outcomes. Research on the applicability of different modelling approaches is ongoing and presented work compares and discusses available models in order to point out their specific strengths and weaknesses in given scenarios. The main aim of this paper is to critically evaluate the main advantages and limitations of available models for large-scale phytoremediation including, among others, the Decision Support System (DSS), Response Surface Methodology (RSM), BALANS, PLANTIX and various regression models. Study compares their applicability and highlight existing gaps in current knowledge with a special reference to improving the efficiency of large-scale phytoremediation of sites contaminated with heavy-metals. The presented work can serve as a useful tool when choosing the most suitable model for the phytoremediation of contaminated sites. (C) 2020 Elsevier Ltd. All rights reserved

Modeling and optimizing the removal of cadmium by Sinapis alba L. from contaminated soil via Response Surface Methodology and Artificial Neural Networks during assisted phytoremediation with sewage sludge

ACLThe study was aimed to model and optimize the removal of cadmium from contaminated post-industrial soil viaSinapis albaL. by comparing two modeling approaches: Response Surface Methodology (RSM) and Artificial Neural Networks (ANN). The experimental design was done using the Box-Behnken Design method. In the RSM model, the quadratic model was shown to predict the closest results in comparison to our experimental data. For ANN approach, a two-layer Feed-Forward Back-Propagation Neural Network model was designed. The results showed that sewage sludge supplementation increased the efficiency of theSinapis albaplant in removing Cd from the soil. After 28 days of exposure, the removal rate varied from 10.96% without any supplementation to 65.9% after supplementation with the highest possible (law allowed) dose of sewage sludge. The comparison proved that the prediction capability of the ANN model was much higher than that of the RSM model (adjustedR-square: 0.98, standard error of the Cd prediction removal: 0.85 +/- 0.02). Thus, the ANN model could be used for the prediction of heavy metal removal during assisted phytoremediation with sewage sludge. Moreover, such approach could also be used to determinate the dose of sewage sludge that will ensure highest process efficiency

Effects of sewage sludge supplementation on heavy metal accumulation and the expression of ABC transporters in Sinapis alba L. during assisted phytoremediation of contaminated sites

ACLATP binding cassette (ABC) transporters, types C, G, and B were monitored via qPCR in order to investigate the influence of heavy metal (HM) contamination of post-industrial and post-agricultural soils and the effects of its supplementation with sewage sludge, on Sinapis alba plants. Five house-keeping genes were selected and validated to ensure the best reference points. The relative expression of ABC types C and G genes was profoundly affected by experimental conditions and included their upregulation after plants exposure to heavy metals and downregulation after supplementation with sewage sludge. However, ABC type C was more responsive then type G. The experimental conditions altered the expression of ABC type C gene faster than ABC type G and thus, the expression of ABC type C can therefore potentially be used as a bioindicator during assisted phytoremediation of degraded sites. In clean soil, supplementation with sewage sludge with a slight content of heavy metals still caused an upregulation in the expression of ABC types C and G, which showed that proper toxicity assessments are necessary to ensure safe application of sewage sludge into soils. Results showed that the analysed genes take a significant part in plants metal detoxification and that their expression is regulated at transcriptional level after exposure to soil contaminated with heavy metals by both, industrial activities and by sewage sludge supplementation. Thus, their expression can potentially be used as an early-warning biomarker when soil supplementation with sewage sludge is incorporated into the soil-management process