Toxicity evaluation of psychotropic pharmaceuticals studying behavioural, physiological and transcriptomic responses of a freshwater snail Radix balthica and a platyhelminthes Schmidtea polychroa

Les médicaments sont fréquement retrouvés dans les effluents de STEP relargués dans l’environnement aquatique. Dans le but de prévenir et de mieux comprendre les impacts des médicaments sur les écosystèmes aquatiques, il semble pertinent d’évaluer les perturbations comportementales, physiologiques et transcriptomiques des psychotropes sur les organismes aquatiques. Dans ce contexte, l’objectif de cette étude a été d’évaluer les perturbations induites par 4 psychotropes (oxazépam, carbamazépine, cyamémazine et sertraline) chez deux organismes, Radix balthica et Schmidtea polychroa. Pour ce faire, des embryons de Radix au stade trochophore et des planaires adultes ont été exposés à chaque psychotrope (du μg/L jusqu’à 100 μg/L). Il en ressort que les psychotropes allongent la durée du développement embryonnaire du Radix et perturbent le déplacement, la reproduction et la digestion, mais pas la régénération de la planaire. D’un point de vue transcriptomique, nous avons réalisé le séquençage du transcriptome en conditions différentielles chez le Radix. Ainsi, nous avons obtenu d’une part les séquences du transcriptome et d’autre part, après analyse en différentiel, 144 contigs différentiellement exprimés par l’oxazépam parmi lesquels 94 ont été vérifiés en RT-qPCR chez des Radix exposés à chaque psychotrope. Il ressort de cette analyse que les psychotropes impactent principalement la voie de signalisation Notch, mais aussi les voies de biosynthèse des polyamines et des catécholamines. Les psychotropes modulent aussi l’expression de gènes codant des protéines de la Matrice Extra Cellulaire (MEC), comme la Matriline ou encore la Dentine sialophosphoprotéine. Chez la planaire, nous avons analysé l’expression de 87 gènes impliqués dans différentes fonctions. Il ressort de cette étude que les 4 psychotropes modulent l’expression de nombreux gènes impliqués dans la mobilité ciliaire et musculaire et dans les systèmes nerveux, reproducteur, excréteur et digestif.Pharmaceuticals are often found in WWTP effluents realesed in surface water. In order to prevent and to understand the pharmaceuticals impact on aquatic ecosystems, it seems relevant to evaluate behavioural, physiological and transcriptomic disturbances of psychotropics on freshwater organisms. The aim of this study was thus to analyse disturbances of 4 psychotropics (oxazepam, carbamazepine, cyamemazine and sertraline) on 2 freshwater organisms, Radix balthica and Schmidtea polychroa. In our experiments, both Radix embryos at the trochophore stage and mature planarian were exposed to each psychotropic (from 1 to 100 μg/L). This psychotropic exposure results in an increase of the duration of Radix embryonic development and a disturbance of the mobility, the reproduction and the digestion but not the regeneration of planarian. Regarding the transcriptomic impact, we performed RNA sequencing in differential conditions of Radix embryos exposed or not to oxazepam. On one hand, this analysis allowed us to obtain the transcriptome sequences and, on the other hand, 144 contigs differentially expressed among which 94 genes were verified by RT-qPCR. The results showed that psychotropics impact mainly the Notch signalling pathways, but also the biosynthetic pathways of polyamines and catecholamines. Psychotropics also disturb the gene expression encoding some Extra Cellular Matrix (ECM) protein such as Matrilin and Dentin sialophosphoprotein. Regarding the molecular study of the psychotropics impact on planarian, we analysed the expression of 87 genes involved in different functions. The results showed that the 4 psychotropics modulate expression of genes involved in ciliary and muscular motility and in the nervous, reproductive and excretory systems. Genes from the digestive system are also impacted by the psychotropics. All the observed impacts on the 2 organisms suggest a possible disturbance on the population fitness and therefore on the freshwater ecosystems

Évaluation de la toxicité de molécules médicamenteuses par une étude des réponses comportementales, physiologiques et transcriptomiques d’un mollusque dulçaquicole (Radix balthica) et d’un plathelminthe (Schmidtea polychroa)

Pharmaceuticals are often found in WWTP effluents realesed in surface water. In order to prevent and to understand the pharmaceuticals impact on aquatic ecosystems, it seems relevant to evaluate behavioural, physiological and transcriptomic disturbances of psychotropics on freshwater organisms. The aim of this study was thus to analyse disturbances of 4 psychotropics (oxazepam, carbamazepine, cyamemazine and sertraline) on 2 freshwater organisms, Radix balthica and Schmidtea polychroa. In our experiments, both Radix embryos at the trochophore stage and mature planarian were exposed to each psychotropic (from 1 to 100 μg/L). This psychotropic exposure results in an increase of the duration of Radix embryonic development and a disturbance of the mobility, the reproduction and the digestion but not the regeneration of planarian. Regarding the transcriptomic impact, we performed RNA sequencing in differential conditions of Radix embryos exposed or not to oxazepam. On one hand, this analysis allowed us to obtain the transcriptome sequences and, on the other hand, 144 contigs differentially expressed among which 94 genes were verified by RT-qPCR. The results showed that psychotropics impact mainly the Notch signalling pathways, but also the biosynthetic pathways of polyamines and catecholamines. Psychotropics also disturb the gene expression encoding some Extra Cellular Matrix (ECM) protein such as Matrilin and Dentin sialophosphoprotein. Regarding the molecular study of the psychotropics impact on planarian, we analysed the expression of 87 genes involved in different functions. The results showed that the 4 psychotropics modulate expression of genes involved in ciliary and muscular motility and in the nervous, reproductive and excretory systems. Genes from the digestive system are also impacted by the psychotropics. All the observed impacts on the 2 organisms suggest a possible disturbance on the population fitness and therefore on the freshwater ecosystems.Les médicaments sont fréquement retrouvés dans les effluents de STEP relargués dans l’environnement aquatique. Dans le but de prévenir et de mieux comprendre les impacts des médicaments sur les écosystèmes aquatiques, il semble pertinent d’évaluer les perturbations comportementales, physiologiques et transcriptomiques des psychotropes sur les organismes aquatiques. Dans ce contexte, l’objectif de cette étude a été d’évaluer les perturbations induites par 4 psychotropes (oxazépam, carbamazépine, cyamémazine et sertraline) chez deux organismes, Radix balthica et Schmidtea polychroa. Pour ce faire, des embryons de Radix au stade trochophore et des planaires adultes ont été exposés à chaque psychotrope (du μg/L jusqu’à 100 μg/L). Il en ressort que les psychotropes allongent la durée du développement embryonnaire du Radix et perturbent le déplacement, la reproduction et la digestion, mais pas la régénération de la planaire. D’un point de vue transcriptomique, nous avons réalisé le séquençage du transcriptome en conditions différentielles chez le Radix. Ainsi, nous avons obtenu d’une part les séquences du transcriptome et d’autre part, après analyse en différentiel, 144 contigs différentiellement exprimés par l’oxazépam parmi lesquels 94 ont été vérifiés en RT-qPCR chez des Radix exposés à chaque psychotrope. Il ressort de cette analyse que les psychotropes impactent principalement la voie de signalisation Notch, mais aussi les voies de biosynthèse des polyamines et des catécholamines. Les psychotropes modulent aussi l’expression de gènes codant des protéines de la Matrice Extra Cellulaire (MEC), comme la Matriline ou encore la Dentine sialophosphoprotéine. Chez la planaire, nous avons analysé l’expression de 87 gènes impliqués dans différentes fonctions. Il ressort de cette étude que les 4 psychotropes modulent l’expression de nombreux gènes impliqués dans la mobilité ciliaire et musculaire et dans les systèmes nerveux, reproducteur, excréteur et digestif

Assessment of Lemna minor (duckweed) and Corbicula fluminea (freshwater clam) as potential indicators of contaminated aquatic ecosystems: responses to presence of psychoactive drug mixtures

International audienceThe pharmaceutical products are emerging pollutants continuously released into the environment, because they cannot be effectively removed by the wastewater treatment plants. In recent years, questions have been raised concerning the environmental risks related to these pollutants. The goal of this research was to evaluate the responses in Lemna minor after 7 days and in Corbicula fluminea after differing durations (1, 3, 7, and 19 days) of exposure to the psychoactive drug mixture (valproic acid, citalopram, carbamazepine, cyamemazine, hydroxyzine, oxazepam, norfluoxetine, lorazepam, fluoxetine, and sertraline) in different concentrations (0, 0 + ethanol, drug concentration (DC) 1 = river water concentration, DC2 = effluent concentration, and DC3 = 10× effluent concentration). In this aim, growth parameters of L. minor, gluthathione S-transferase (GSTs), catalase (CAT), ethoxyresorufin-O-deethylase (EROD) and/or gene expressions (pi-gst, cat, cytochrome P450 4 (cyp4), multidrug resistant 1 (mdr1), and superoxide dismutase (sod)) were measured. GST activities increased significantly in L. minor exposed to DC3, but no changes were found in CAT activity. In C. fluminea, EROD activity was induced significantly in both gill and digestive gland tissues after 3 days’ exposure to DC3, while a GST increase was observed only in digestive gland tissues, suggesting that these pharmaceuticals induced an oxidative effect. Gene expression analysis revealed transient transcriptomic responses of cyp4, sod, and mdr1 under drug concentrations 2 or 3 and no change of expression for the other genes (cat and pi-gst) or condition (environmental drug concentration) tested. Finally, the data reported in this study represent important ecotoxicological information, confirming that this enzyme family (cyp4, sod, and mdr1) may be considered as a sensible and early indicator of exposure to drugs and emphasizing the involvement of selected genes in detoxification pathways

Evaluation of psychiatric hospital wastewater toxicity: what is its impact on aquatic organisms?

International audienceThe primary source of pharmaceuticals to the aquatic environment is the discharge of wastewater effluents. Pharmaceuticals are a large and diverse group of compounds. Among them, psychotropic substances are particularly interesting to study due to their specific known mode of action. The present study was performed to investigate the effects of wastewater effluents from a psychiatric hospital wastewater treatment plant (WWTP) on several aquatic organisms. All the analyzed pharmaceuticals (10 compounds) were detected in WWTP effluents as well as in the receiving river. Although the environmental concentrations were generally at trace levels (ng L-1 to μg L-1), induce toxic effects were observed. This study showed the effects of the WWTP effluents on the oogenesis and/or embryogenesis of amphipod crustacean Gammarus fossarum, Japanese fish medaka Oryzias latipes, mollusk Radix peregra, and planarian Schmidtea polychroa. A decrease of the number of oocytes and produced embryos was observed for G. fossarum and S. polychroa. Similarly, the hatching rate of R. peregra was affected by effluents. In the receiving river, the macroinvertebrate community was affected by the wastewater effluents discharge