SETAC EUROPE 32
NDANNUAL MEETING
15–19 MAY 2022 | COPENHAGEN, DENMARK
“TOWARDS A REDUCED POLLUTION SOCIETY”
ABSTRACT BOOK
Abstract Book
SETAC Europe 32
ndAnnual Meeting
Table of Contents
About SETAC ...3
Abstracts ...5
Track 1: Ecotoxicology and human toxicology: From molecules to organisms, from omics to in vivo ...5
Track 2: Ecotoxicology becomes stress ecology: From populations to ecosystems and landscapes ...93
Track 3: Environmental chemistry and exposure assessment: analysis, monitoring, fate and modeling .167 Track 4: Ecological risk assessment and human health risk assessment of chemicals, mixtures and stress- ors and risk mitigation strategies ... 410
Track 5: Life Cycle Assessment and foot-printing ...540
Track 6: Environmental policy, risk management, and science communication ...595
Track 7: Think-outside-the-box (fundamentally new concepts, innovative and controversial ideas, and interdisciplinary issues) ...648
Track 8: Special Sessions ...662
Author Index ... 717
Affiliation Index ...724
This book compiles the abstracts from the 32
ndannual meeting of the Society of Environmental Toxicology and Chemistry – Europe (SETAC Europe), conducted from 15–19 May 2020 in Copenhagen, Denmark, and online.
The abstracts are reproduced as submitted by the author and accepted by the scientific Committee. They
appear in order of abstract code and alphabetical order per presentation type. The poster spotlight abstracts
are included in the list of poster abstracts. The presenting author of each abstract is underlined.
No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, electrostatic, magnetic tape, mechanical, photocopying, recording, or otherwise, without permission in writing from the copyright holder. SETAC Europe’s consent does not extend to copy- ing for general distribution, for promotion, for creating new works, or for resale. Specific permission must be obtained in writing from SETAC for such copying. Direct all inquiries to SETAC Europe.
PRINT ISSN 2309-8031 - ONLINE ISSN 2310-3043 © 2022
Society of Environmental Toxicology and Chemistry Europe (SETAC Europe)
About SETAC
In the 1970s, no forum existed for interdisciplinary communication among environmental scientists, biologists, chemists, toxicologists, managers, engineers or others interested in environmental issues. The Society of Environmental Toxicology and Chemistry (SETAC) was founded in North America in 1979 to fill the void and quickly saw dynamic growth in the Society’s membership, meeting attendance and publications.
A unique strength of SETAC is its commitment to balance the scientific interests of government, aca- demia and business. The Society by-laws mandate equal representation from these three sectors for officers of the World Council and Geographic Unit Boards of Directors and Councils, and in the composi- tion of committees and other society activities. The proportion of members from each of the three sectors has remained nearly equal over the years.
The Society is concerned about global environmental issues. Its members are committed to
Environmental Quality Through Science®, timely and effective communication of research, and inter- actions among professionals so that enhanced knowledge and increased personal exchanges occur.
Therefore, SETAC publishes two globally esteemed scientific journals and convenes annual meetings around the world, showcasing cutting-edge science in poster and platform presentations. Because of its multidisciplinary approach, the scope of the science of SETAC is broader in concept and application than that of many other societies.
SETAC’s growth is reflected in the founding of Geographic Units around the world. SETAC Europe was established in 1989 as an independent organisation, followed by SETAC Asia-Pacific in 1997 and SETAC Latin America in 1999. In 2002, the four existing organisations joined together under the governance of the SETAC World Council. SETAC Africa is the most recent Geographic Unit, which was adopted in 2012. As evidence of international acceptance of the SETAC model and of the great interest at the local level, regional chapters and branches have emerged in a number of countries.
SETAC publishes two journals, Environmental Toxicology and Chemistry (ET&C) and Integrated Environmental Assessment and Management (IEAM). ET&C is dedicated to furthering scientific knowl- edge and disseminating information on environmental toxicology and chemistry, including the application of these sciences to risk assessment. Integrated Environmental Assessment and Management focuses on the application of science in environmental decision-making, regulation and management, including aspects of policy and law, and the development of scientifically sound approaches to environmental prob- lem solving. Together, these journals provide a forum for professionals in academia, business, government and other segments of society involved in the use, protection and management of the environment for the enhancement of ecological health and human welfare.
SETAC books provide timely in-depth reviews and critical appraisals on scientific subjects relevant to understanding a wide range of contemporary topics pertaining to the environment. These include any aspect of environmental chemistry, toxicology, risk assessment, risk management or environmental policy.
SETAC has two administrative offices, in Pensacola, Florida, USA, established in 1992, and in Brussels, Belgium, established in 1993.
www.setac.org
Environmental Quality Through Science ®
Track 1: Ecotoxicology and human toxicology: From molecules to organisms, from omics to in vivo
1.01 Advantages of using lab and field collected invertebrates in ecotoxicology: Challenges and opportunities for Environmental Risk Assessment (Part I)
1.01.T-01 Protein Binding on Acutely Toxic and Non-Toxic Polystyrene Nanoparticles During Filtration by Daphnia magna
Ms. Egle Kelpsiene1, Irene Brandts2, Katja Bernfur1, Mikael T. Ekvall1, Mariana Teles2 and Tommy Cedervall1, (1)Lund University, Sweden, (2)Universidad Autonoma de Barcelona, Spain
Nanomaterials can adsorb biomolecules to their surface and form a protein corona. In the present study, we investigated the protein profile bound to different sizes of aminated and carboxylated polystyrene (PS) nanoparticles after passing through the digestive tract of the freshwater zooplankton Daphnia magna. We found that acutely toxic aminated 53 nm PS nanoparticles binds a different set of proteins compared to other non-toxic PS nanoparticles. A higher amount of proteins appeared to be bound to the aminated PS nanoparticles. The fact that the aminated nanoparticles bound a larger number of proteins, agrees with the aggregate size results. The protein bound to toxic nanoparticles can be divided into two groups. One group of proteins which function is related to the digestive system, whereas the other group of proteins can be related to the epithelium, intracellular structures, and processes. Finally, we observed that not only proteins bind to surfaces of the nanoparticles. Triglycerides effectively bind to 200 nm carboxylated PS nanoparticles but not to the other tested nanoparticles. These results provide information about the
composition of the corona formed on surfaces of nanoparticles after short-term (4 h) incubation with D. magna.
1.01.T-02 High-Throughput Screening of Phototactic Behaviour in Daphnia magna: From Vertical to Horizontal Responses to Light
Carlos Barata Dr1, Marina Bellot2 and Cristian Gómez-Canela3, (1)CSIC, Spain, (2)University Ramon Llull IQS, Spain, (3)School of Engineering, Institut Químic de Sarrià-Universitat Ramon Llull, Spain
Animal behavioural trait responses are increasingly being used in environmental risk assessment. Nevertheless, behavioural responses are still hampered by a lack of ecological relevance and standardisation. This is the case for diel vertical migration of zooplanktonic species. Previously we develop a vertical oriented behavioural hardware able to reproduce phototactic fish induced depth selection in Daphnia magna and its modulation by fish kairomones and psychotropic drugs. This study aims to test if it is possible to use an horizontal 24 multi-well plate maize set up to assess phototactic fish induced responses in D. magna. The study was conducted using two clones with opposed phototaxis upon exposure to fish kairomones and using psychotropic drugs known to modulate phototaxis. Acrylic strips opaque to visible light but not to the infrared one were used to cover half of the arena of each of the wells of the multi-well plate. Clone P132,85 showed positive phototaxis in either the vertical and horizontal set up and negative phototaxis when exposed to fish kairomones or to the muscarinic acetylcholine receptor antagonists scopolamine and atropine. The opposite behavior was observed for clone F. Diazepam and pilocarpine ameliorate fish kairomone induced negative phototaxis and picrotoxin increased it only on clone P132,85 in the vertical set up. The study of neurotransmitter metabolites showed much greater concentrations of dopamine and of glycine in clone F, which may be relate to its negative phototaxis and its observed lower responsiveness to fish kairomones. The results from this study suggest a simple, fast, high throughput assay for D.
magna, and gains insight on the impacts of natural and psychotropic compounds on the swimming behaviours of a model crustacean species used in ecotoxicology studies.
1.01.T-03 Differential Sensitivity and Synergy in Adult and Larvae Lepidoptera
Claire Badder, Pollution, UK Centre for Ecology and Hydrology, Wallingford, United Kingdom
Differential sensitivity to pesticides between insects may have many biological explanations. One such explanation may be the potential of a species to metabolise the toxicant. Therefore, an awareness of the presence of metabolic enzymes such as cytochrome P450’s (CYPS), glutathione-S-transferases (GSTs) and carboxylesterases in a specific species may hold an insight into the potential sensitivity or tolerances the species is likely to have towards chemical toxicants . Here we aim to we determine the toxicity of the insecticide cypermethrin and the fungicides azoxystrobin, chlorothalonil and prochloraz to adult and larval stages of the cabbage moth (Mamestra brassicae). Using single chemical toxicity data and TKTD modelling, we aim to identify cases of synergism between cypermethrin and the fungicides in binary mixtures. Finally, we begin to explore any differences in sensitivity using the levels of expression of CYPS, GSTs and carboxylesterases in the transcriptome of the larvae and adult. Single chemical tests were analysed using the GUTS_RED_SD model and revealed fungicides had no effect on mortality on cabbage moth larvae. Therefore, no LC??s could be calculated for azoxystrobin, chlorothalonil or prochloraz. However, the insecticide cypermethrin had an LC?? of 46.3 and 11.6 at 48 and 96 hours respectively. This differed in adult moths as there was a greater tolerance to cypermethrin with LC?? values of 90.5 at 48 hours and 22.7 and 96 hours, a difference of almost 50%. Larval mixture testing identified slight synergy in larvae between cypermethrin and prochloraz and a more pronounced synergistic interaction between cypermethrin and chlorothalonil. No synergism was seen between azoxystrobin and cypermethrin. Adult mixture tests are on-going but it is hypothesised that synergistic effects will be lessened as is seen in the single chemical cypermethrin test. Further, a catalogue was created of CYPS, GST’s and carboxylesterases from the transcriptome of cabbage moth larvae. The adult moth transcriptome is underway and will be compared against the larval transcriptome. We hypothesise that differences in sensitivity may begin to be explained by differences in expression of metabolic enzymes. We hope that this multi-faceted approach of lab exposure, GUTS modelling and transcriptomics provides an insight into the power of omics as a predictive tool of toxicity, particularly when used alongside traditional toxicity testing.
1.01.T-04 Experimenting the Plasticity of Sensitivity From Field-Populations of Gammarus fossarum in Response to Chronic Cadmium Contamination: Tolerance Acquisition, Transfer and Reversibility
Auréline Lalouette1, Davide Degli Esposti2, Laura Garnero3 and arnaud chaumot4, (1)INRAE Centre de Lyon-Grenoble Auvergne-Rhône-Alpes, France, (2)INRAE, France, (3)INRAE Lyon, France, (4)Irstea, France
Chronic exposure to metal contamination can induce changes in the sensitivity of populations. An increase in tolerance,
transmissible between generations, can be observed in certain cases without being due to genetic adaptation. This particular case of transgenerational acclimatization using non-genetic mechanisms of heredity has been documented in natural populations of the crustacean Gammarus historically exposed to metallic geochemical background. In these populations, the acquisition of cadmium (Cd) tolerance has been shown to be consistent with the manifestation of parental effects of exposure. The objectives of our study were then to confirm the existence of Cd tolerance among several G. fossarum populations at a regional scale, to better understand the nature of this tolerance acquisition (transitory acclimatation or permanent acquisition of tolerance by the organisms) and to study the mechanisms of transmission between generations. For that, natural Cd-contaminated and uncontaminated field G.
fossarum populations was used in three laboratory protocols, 1- the returning in clean water in the lab of Cd-contaminated and uncontaminated field G. fossarum populations to question the reversible nature of sensitivity, 2- the re-matching spawners from Cd-tolerant and sensitive field populations to study the transfer of the tolerance to progeny and, 3- the etablishment of a Cd chronic exposure in the lab at environmentally relevant concentrations to test the mechanisms seen in naturally tolerant populations. This study confirm the existence of natural populations capable of developing mechanisms of tolerance in Cd contaminated environments. Moreover, our results show that tolerance is not fixed for the life time of individuals, not depending therefore on genetic mechanisms or developmental plasticity, while this tolerance can be transmitted between generations by parental effects with a symmetrical role of the two sexes. Thus, the use of invertebrates is of real interest to increase our understanding of long-term effects of chimical contaminations in ecosystems by allowing to conduct environmentally relevant ecotoxicological studies.
1.01.T-05 Predicting the Survival of Field Gammarids Provides Additional Insights on the Impacts of Fluctuating Pesticide Exposures
Dr. Maricor Jane Arlos1, Andreas Focks2, Juliane Hollender3 and Christian Stamm4, (1)University of Alberta, Edmonton, AB, Canada, (2)University of Osnabruck, Germany, (3)Eawag - Swiss federal Institute of Aquatic Science and Technology, Switzerland, (4)Eawag, Swiss Federal Institute of Aquatic Science and Technology, Switzerland
The analytical detection and modelling approaches to assess pesticide exposure in various environmental compartments have substantially improved over the last several decades. However, the threshold values considered in the current regulatory risk assessment are still based on laboratory studies that do not mirror the conditions captured in chemical monitoring and modelling.
In this study, we employed a toxicokinetics-toxicodynamic (TKTD) model from the General Unified Threshold model of Survival (GUTS) framework to predict the impact of fluctuating pesticide concentrations on the survival of gammarids (Gammarus pulex) in an agricultural stream located in Switzerland. Individual exposure to azoxystrobin, chlorpyrifos and diazinon were predicted to have an acute and/or chronic impact on the survival of gammarids. In contrast, exposure to imidacloprid and propiconazole were predicted to have a low impact as their environmental concentrations must be multiplied by more than 100 before a survival probability of 50% is observed. Our modelling approach subsequently revealed that the duration of exposure drives the effects more than the maximum concentration as typically considered in traditional risk assessments. We further extended the GUTS model to incorporate mixture effects using concentration addition. We found that a considerable impact is expected when
gammarids are exposed to organophosphates (chlorpyrifos, chlorpyrifos-methyl, and diazinon), whereas a mild impact is predicted for neonicotinoids (imidacloprid and thiacloprid). Given that the gammarids are exposed to many other environmental
contaminants and stressors, a decline in their survival probabilities because of chemical stress will likely influence their individual fitness and/or overall population viability. Additional laboratory and/or field studies are required to validate the mixture
assumptions in our model. However, by incorporating the temporal aspects of environmental exposure and the associated consequences on survival, our study added a layer of realism that improves the current risk assessment of pesticides.
1.01 Advantages of using lab and field collected invertebrates in ecotoxicology: Challenges and opportunities for Environmental Risk Assessment (Part II)
1.01.T-06 Biomarker Development in High-Resolution Pesticide Mixture Exposures Using the Springtail Folsomia candida Ruben Rob Bakker1, Yuliya Shapovola2, Tjeerd Dijkstra3, Dick Roelofs4, Tom Heskes5, Cornelis A.M. (Kees) van Gestel6 and Katja Hoedjes7, (1)Animal Ecology, Vrije Universiteit Amsterdam, Amsterdam, Netherlands, (2)Radboud University Nijmegen, Nederland, (3)University Clinic Tübingen, Germany, (4)Vrije Universiteit, Netherlands, (5)Radboud University Nijmegen, Netherlands, (6)Vrije Universiteit Amsterdam, Netherlands, (7)Vrije Universiteit, Nederland
Invertebrate populations worldwide are in decline, at least partly due to intensive pesticide application. Traditionally, pesticide risk assessment has been based on the results of standardized tests measuring the effects of a single pesticide on the reproduction or survival of model organisms. It is difficult to extrapolate these findings to field conditions as most agricultural soils are polluted by pesticide mixtures and the effects of their (synergistic) interactions on invertebrates are largely unknown. Gene regulatory patterns could be used as biomarkers to determine the type of soil pollution present and its intensity under varying mixture compositions. Generating high-throughput gene expression data of high-resolution exposure intensities, and the development of methods to analyse this data are crucial for linking gene-expression patterns to adverse effects on reproduction, and for subsequent implementation of biomarkers in risk assessment or biomonitoring. In this study, transcriptomic data was obtained for the soil ecotoxicological model species Folsomia candida (springtails) under a high-resolution grid-design-exposure of either two
neonicotinoids, imidacloprid and clothianidin, or imidacloprid and cyproconazole, a fungicide. In total the exposure consisted of 168 samples over 25 unique conditions in two experiments . Obtained gene expression data on all pesticides was analysed simultaneously with a joint Gaussian processes (GP) additive model, as GP-models allow for the analysis of non-linear
differential gene expression patterns in a grid design. In total, this method identified 2049 differentially expressed genes (DEGs) under the exposure of neonicotinoids and 1080 DEGs under the exposure of cyproconazole. Gene set enrichment (GSE) analysis reveals shared and distinctive gene functions for DEGs under neonicotinoid and cyproconazole exposure, which can be used to identify points of synergistic interaction or used for biomarker development respectively. The candidate biomarkers are identified from distinctive GSE terms and validated on agricultural soils, providing risk assessors with metrics of adverse effects of pesticide presence that can be used on field-relevant mixture exposures.
1.01.T-07 Functional Transcriptomic Fingerprints of Neurotoxic Modes-Of-Action in Daphnia magna
Julia Pfaff1, Hannes Armin Reinwald2, Dr. Steve Uwa Ayobahan2, Julia Alvincz2, Dr. Bernd Göckener3, Orr Shomroni4, Gabriela Salinas4, Rolf-Alexander During5, Christoph Schaefers2 and Sebastian Eilebrecht2, (1)Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Germany, (2)Fraunhofer IME - Institute for Molecular Biology and Applied Ecology, Germany, (3)Fraunhofer IME - Institute for Molecular Biology and Applied Ecology, Schmallenberg, Germany, (4)NGS - Integrative Genomics Core Unit, Germany, (5)Justus Liebig University Giessen, Germany
Active ingredients of pesticides, biocides or pharmaceuticals can cause undesirable side effects in the aquatic ecosystem.
Therefore, an environmental hazard and risk assessment is required prior to the registration of new active substances. The freshwater crayfish Daphnia magna is a model organism for evaluating acute and chronic toxicity in aquatic invertebrates.
However, standardized tests using Daphnia are limited to immobility and reproductive endpoints and thus provide limited insight into underlying modes-of-action (MoA). Here, we applied transcriptome profiling to a modified acute immobilization assay of D.
magna to analyze and compare gene expression profiles induced by the GABA-gated chloride channel blocker fipronil and the nicotinic acetylcholine receptor (nAChR) agonist imidacloprid. To detect compound-induced gene expression changes at low effect concentrations, an acute immobilization test with D. magna was performed following the corresponding OECD guideline test. 48 hours after test initiation, total RNA was extracted from the daphnids and purified polyA-RNA was subjected to RNA sequencing for differential gene expression analysis. The observed signatures for fipronil and imidacloprid were readily
distinguishable between the two compounds at the gene level without major overlap. While fipronil exposure affected a number of metabolic pathways, including ATP synthesis and cholesterol biosynthesis, metabotropic glutamate receptor signaling, and a pathway associated with Alzheimer's disease - the presenilin pathway in mammals, impaired imidacloprid pathways associated with Parkinson's disease, mammalian vasopressin synthesis, GABA synthesis, and aminobutyrate degradation. Our study suggests that the integration of transcriptomics into a modified version of the acute immobilization assay of D. magna can be used to differentiate modes-of-action of test compounds of interest at low effect concentrations. Such a combination of systems biology methods with ecotoxicological test guidelines will help to understand the underlying molecular mechanisms of adverse effects on invertebrate organisms and populations, which will be important for ecotoxicological hazard assessment.
1.01.T-08 Physiological Colour Changes in Ecotoxicology
Thomas Knigge1, Anaid Gouveneaux2, Apolline Chabenat3, Eleanor Feuerhelm4, Sarah Hue5, Jeanne Legendre5, Tiphaine Monsinjon5, Alex Ford4 and Cécile Bellanger6, (1)Universite du Havre, France, (2)Université de Caen Normandie, France, (3)University Le Havre Normandie, France, (4)University of Portsmouth, United Kingdom, (5)Université Le Havre Normandie, France, (6)UniversitÈ de Caen Normandie, France
Many animals display rapid colour changes for the purpose of camouflage. Physiological colour changes can be controlled either nervously, as in cephalopods, or hormonally, as in crustaceans. Both, neuronal and neurohormonal control mechanisms are regulated by neurotransmitters. In cephalopods, serotonin induces the relaxation of the muscle crown leading to pigment concentration in the chromatophores. In crustaceans, dopamine stimulates pigment concentration via red pigment concentrating hormone, whereas serotonin ensures pigment dispersion via pigment dispersing hormone. Environmental pollutants can
potentially interfere with these mechanisms of physiological colour change and cryptic behaviour. We exposed Carcinus maenas, Crangon crangon and Sepia officinalis to waterborne selective serotonin and/or norepinephrine reuptake inhibitors, which are known to alter the levels of serotonin and other biogenic amines in wildlife. Venlafaxine provoked pigment concentration in chromatophores of S. officinalis at the same concentration (10-4M) as did serotonin. Fluoxetine alone (5 ng L–1) or in combination with venlafaxine (2.5 or 5 ng L–1) improved uniform, but not disruptive body patterns in newly hatched cuttlefish. C. crangon exposed to 10-1000 ng L–1 fluoxetine for up to one week showed a significant increase in darkening following 30 min on black substrates. Similarly, pigment dispersion in juvenile C. maenas transferred from a white to a dark background tended to increase when exposed to fluoxetine at 5 ng L–1, but when exposed to a combination of fluoxetine and venlafaxine at 5 ng L–1 each, darkening was significantly impaired. These studies demonstrate that physiological colour changes are susceptible to
environmental chemicals that interfere with the neuronal or neurohormonal control of chromatophore movements. Hence, they may serve as indicators of environmental pollution and could point to detrimental effects on behavioural ecology that are important for the animals’ survival. Colour change bioassays, therefore, provide a promising tool for environmental testing with the advantage of being non-invasive, inexpensive and relatively straightforward.
1.01.T-09 Organophosphate Pesticides in Aquatic Ecosystems: Study CASE on Chironomus riparius Impact at MOLECULAR Level
Ana-Belen Muniz-Gonzalez1, Valeria Lencioni2 and Jose-Luis Martinez-Guitarte, PhD3, (1)National Distance Education University (UNED), Spain, (2)MUSE-Museo delle Scienze Trento, Italy, (3)UNED, Spain
Organophosphate pesticides (OPPs) have gained worldwide concern by their persistent and hazard to the environment, surrounding nearly 40% of the global market and in constant increase reaching easily to aquatic systems acting as main OPPs reservoir. Chlorpyrifos (CPF) insecticide has been the most employed for crops protection in Italy derived in intensify presence in surface waters (0.12-15.38 ng/L) along the country. The harmful CPF impact is widely documented reaching to be banned by EPA in august 2021, although as a semi-persistent pollutant, continue to be present in aquatic systems for years. However, the impact on field organisms has been poorly studied apart from our previous experience on Diamesa species. Therefore, this work is focused to evaluate the CPF impact at molecular level on C. riparius larvae collected in Rio Gola stream (32T 664117 5100540 UTM, October 2020), to check the influence on benthic macroinvertebrates in close contact with polluted sediments. The larvae were exposed at 0.011 µg/L, 0.11 µg/L and 1.1 µg/L for 24 and 72 h, in controlled laboratory conditions (8 light:16 dark hours; 20
°C). Sub-organismal response was evaluated using an array (48 genes) designed for this specie by Real Time PCR. After 24h, Dis, and NHR38 (endocrine system); GSTo1 (detoxification response); Proph (immune system); XRCC1 (DNA repair), and LIP (lipid metabolism) were altered. At 72h higher response was observed JHAMT, Dis, NHR38, NHR96, and FKBP39 (endocrine system);
Cyp4c1, Cyp12a1, GSTd6 (detoxification response); and hsp70 (stress response) modulated. Hormone’s synthesis inhibition confirmed the endocrine disruptor CPF capacity. Moreover, phase II modulation reinforces the role of CPF as oxidative compound, being confirmed by the immune system activation strongly related to ROS. Lipid metabolism was modulated could limit insect’s functions because are essential source of energy (i.e: reproduction). Besides, the DNA damage seems to be drastic at 72h with apoptosis initiation (Decay). Finally, the stress response could indicate defective protein folding. Our findings confirm the risk of CPF, at environmental relevant concentrations, affecting the metabolism and future survival on a key aquatic organism.
The differences in expression observed between 24 and 72h reinforce the importance to check the molecular modulation along the time in above all organisms as C. riparius with short life cycles.
1.01.T-10 How Nano Pesticides Affect Non-Target Organisms: Toxicity of Three Nanopesticides on Daphnia magna Mahleh Eghbalinejad1, Maria del Rocio Lopez-Cabeza2, Jan Kotou?ek3 and Jakub Hofman4, (1)Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Brno, Czech republic, Czech Republic, (2)Masaryk University, Faculty of Science, RECETOX, Spain, (3)Department of Pharmacology and toxicology, Veterinary Research Institute, v.v.i., Brno, Czech Republic, Czech Republic, (4)RECETOX, Faculty of Science, Masaryk University, Czech Republic
Nanopesticides as a potential solution to solve the risk and drawbacks of conventional formulations of pesticides have been attracting attention in both research and market areas. However, the risk of these new formulations still needs to be studied on non-target species and regulated specifically. In this regard, we prepared three nanopesticides composed of nanoparticles of polycaprolactone (PCL), polyhydroxybutyrate (PHB) and nanostructured lipid carrier (NLC) loaded with the fungicide tebuconazole (TBZ (a.i.)), as well as three suspensions of the nanocarriers without a.i. Then, we assessed the toxicity of all the nanoformulations to an aquatic model, Daphnia magna, through immobilization bioassay. For that, in total 28 neonates < 24h old in four replicates were exposed to 5 dilutions of each nanopesticides obtaining a total TBZ concentration of 36, 18, 9, 4.5 and 2.25 µg mL-1. Likewise, the toxicity of the nanocarriers without a.i. was tested as control. The aim is to understand how Daphnia magna behaves in different concentrations of a.i. associated with nanocarriers and in the presence of the nanocarriers without a.i.
over 48h. The total concentrations, release rate of a.i. and the stability of NPs at time 0 and 48h were checked. According to the results, D. magna reacted differently to exposure with each NPs. The dose-response results showed that the EC50 values of all the nanopesticides were high compared to the value for pure TBZ, which shows that the toxicity of all nanopesticides was lower than that of pure TBZ. The toxicity of the NPs decreased depends on the type of nanocarrier as follows: PCL-TBZ< PHB-TBZ< NLC- TBZ. The toxicity of the nanocarrier without a.i. was diverse. As, PCL showed some toxicity while PHB nanocarrier had almost no effect on the immobilization of D. magna. Consequently, since the toxicity of the tested NPs were less than that of pure TBZ, they can be a promising solution to adress the concern regarding the risk of TBZ in one hand and in other hand as they showed improvement of a.i. solubility. These results are the effect of a 48h exposure of D. magna to nanopesticides. Therefore, longer- term exposure and reproduction bioassays are important for robust results, which they are ongoing. In general, as the different nanopesticides and their nanocarriers in the present study showed different responses, it highlights the importance of relevant guidelines for environment risk assessment of nanopesticides specifically.
1.01 Advantages of using lab and field collected invertebrates in ecotoxicology: Challenges and opportunities for Environmental Risk Assessment (Poster)
1.01.P-Mo001 The Comet Assay With Standard Aquatic Test Organisms As an Alternative Test System for an Environmental Risk Assessment of Pharmaceuticals
Marwin Jafari1, Maria Vogt2, Dr. Elke Eilebrecht3, Lena Kosak4, Jasmin Brueckner5, Kathi Westphal-Settele5, Ute Kühnen6 and Karsten Schlich4, (1)Fraunhofe IME, Schmallenberg, Germany, (2)Fraunhofer IME - Institute for Molecular Biology and Applied Ecology, Germany, (3)Ecotoxicology, Fraunhofer IME, Schmallenberg, Germany, (4)Ecotoxicology, Fraunhofer IME - Institute for Molecular Biology and Applied Ecology, Schmallenberg, Germany, (5)Umweltbundesamt / German Environment Agency, Germany, (6)German Environment Agency, Germany
Human pharmaceuticals find their way into aquatic ecosystems e.g. via municipal wastewater, where they pose a potential threat to aquatic organisms. A comprehensive environmental risk assessment (ERA) is necessary to minimize their risks. In accordance to the guidance on human pharmaceuticals for an ERA, effects on aquatic organisms are determined based on standardized guidelines including, among others, the Daphnia magna Reproduction Test (OECD 211). The EMA guideline for ERA was published in 2006 based on the efficacy of active substances in previous years. Meanwhile there are more potent active
substances, putting into question whether the test systems still offer sufficient protection for aquatic organisms. In a previous project (German Environmental Agency, FKZ 3718 65 420 1) alternative test systems for an ERA were identified. One of the identified test systems was the Comet Assay, a genotoxicity test that quantifies DNA damage by measuring the fraction of DNA that migrates out of a nucleus during gel electrophoresis (Tail intensity or TI%), with environmentally relevant organisms.
Therefore, we established the Comet assay with the invertebrate D. magna and tested the effect of several pharmaceuticals either used in oncology (e.g. Cyclophosphamid) or cardiologica (e.g. Edoxaban) to account for genotoxicity as a potential adverse effect of pharmaceuticals. D. magna were exposed for 48 hours in accordance to the OECD 202 with test concentrations chosen to result in sublethal effects. After exposure, immobility was recorded and immobile daphnids were discarded. Daphnia cells were isolated and purified via filtration. The cells were mixed with agarose and pipetted onto microscopy slides. The slides were then incubated in a lysis solution to remove membranes. A treatment in an alkaline solution was performed causing the DNA to unwind and separate into single strands. Gel electrophoresis was performed and the slides were washed and dried overnight. The gels were stained using the DNA dye Vista Green and examined under a fluorescence microscope. Images were evaluated using the
software CaspLab in order to determine tail intensity (TI%) values. The comet assay on D. magna was successfully established. In a calibration with hydrogen peroxide an increase in median TI% of over 60% was determined confirming that the method is capable of detecting genotoxicity. Of the pharmaceuticals tested so far, only Edoxaban showed a dose dependent increase in TI%.
1.01.P-Mo002 Impact of Carbon Nanotubes on an Emerging Cellular Organism: Physarum polycephalum
Manon Ternois1, Audrey Dussutour2 and Emmanuel Flahaut3, (1)CRCA and CIRIMAT, France, (2)CRCA-CBI UMR CNRS 5169 UPS, France, (3)CIRIMAT UMR CNRS 5085 UPS INPT, France
Carbon nanotubes (CNTs) are a category of nanomaterials with applications in all fields (energy, transports, composite materials and environment) due to their exceptional mechanical, optical and electronic properties. However, their use should depend on the non-toxicity of these nanomaterials, and little is still known on their impact on living beings1,3,4. This study evaluated the toxicity of double-walled carbon nanotubes (as a model of CNTs in general) on the unicellular organism Physarum polycephalum, which, due to its simplicity of organization and behavioural complexity2, is a model of choice. The objective was to assess the fate of CNTs within the cell of Physarum polycephalum, and to evaluate their impact on the behaviour of the latter. For this purpose, organisms were contaminated along three routes of exposure (topical, food, environment) in order to determine the internalization of CNTs. The food exposure was the more efficient. Accumulation and excretion of CNTs within the cell were then assessed only by food exposure of the organism. No bioaccumulation and a fast excretion from the cell were observed. From a behavioural point of view, impacts on the migration speed, the fusion process and the cognitive performance were also investigated.
1.01.P-Mo003 Polyethylene Impacts in Fatty Acids and Carbohydrate Profiles of the Estuarine Species Scrobicularia plana
Paola Talarico1, Silvia Lomartire2, Carolina Rocha3, João Marques3 and Ana Marta Goncalves4, (1)MARE - Marine and Environmental Sciences Centre, University of Coimbra, Pianopoli, Italy, (2)MARE -Marine and Environmental Sciences Centre, Faculty of Sciences and Technology, University of Coimbra, Italy, (3)University of Coimbra, MARE - Marine and Environmental Sciences Centre, Portugal, (4)MARE - Marine and Environmental Sciences Centre, University of Coimbra, Coimbra, Portugal Microplastic (MP) pollution in marine environments is a topic of emerging concern. Due to their ubiquity and small size, these particles are easily internalized by marine organisms, causing adverse biological responses. Bivalves are considered bioindicators of environmental health due to their wide distribution, ability to filter large volumes of water, sessile behaviour and ecological and commercial importance. Biomarkers, such as fatty acids (FAs) and carbohydrates (CHs), have been recently proven to be
powerful tools to detect and evaluate an organism response to contaminants’ exposure. The present study assesses the biochemical (FAs and CHs content) and ecotoxicological responses of two size classes (big and small) of the estuarine bivalve Scrobicularia plana exposed to cospheric polyethylene (PE) microparticles (75-90 ?m). Organisms were exposed to environmental relevant and also higher concentrations of virgin PE microparticles (30 particles L-1, 90 particles L -1, 150 particles L-1, 450 particles L-1) for 96h. Results showed that PE ingestion occurred at all concentrations and microparticles accumulated in the visceral mass of S.
plana, with the small size being the most affected class. PE exposure caused significant decreases in polyunsaturated fatty acids (PUFAs) content, including some essential fatty acids (EFAs), which are important compounds for their growth, reproductive ability and overall fitness. Concerning CHs results, PE exposure significantly decreased xylose, galactose and glucose contents.
The decrease in glucose content can be interpreted as an exploitation of glycogen reserves for glucose availability caused by the condition of stress induced by the presence of the pollutant. The decrease in long-chain fatty acids, such as PUFAs, might be a consequence of depletion of glycogen reserves. As available glucose declines, S. plana started exploiting other energy sources to compensate such loss of energy, triggering lipids hydrolysis. These alterations may affect the overall fitness of the organisms and may cause changes in their nutritive value and thus along the trophic food chain, with potential implications for human health. As bivalves are consumed without removal of the digestive tract, they represent an important route of MP exposure for humans and ecotoxicological data on the effects of MPs are urgently needed as potential consequences on human health are still unknown.
1.01.P-Mo004 Polyethylene Impacts on the Antioxidant Defence System of the Estuarine Species Scrobicularia plana Mr. Jorge Gutierrez Serrano1, Andreia Filipa Mesquita2, Silvia Lomartire3, João Marques4 and Ana Marta Goncalves5, (1)MARE - Marine and Environmental Sciences Centre, University of Coimbra, Las Rozas de Madrid, Spain, (2)CESAM &
University of Aveiro, Portugal, (3)MARE -Marine and Environmental Sciences Centre, Faculty of Sciences and Technology, University of Coimbra, Italy, (4)University of Coimbra, MARE - Marine and Environmental Sciences Centre, Portugal, (5)MARE - Marine and Environmental Sciences Centre, University of Coimbra, Coimbra, Portugal
Plastics have become a substantial part of modern life and ubiquitous in all environments. Microplastics (MPs) are emerging
contaminants in marine environments and pose a serious threat to the marine ecology. MPs arise from land sources, deterioration of larger plastics, or intentionally manufactured as “primary” or “virgin” microbeads. MPs can be found in diverse shapes and sizes, and their long-term effect is yet unknown. Estuarine ecosystems, as transitional systems between marine, fluvial, and terrestrial environments, represent a hotspot for MPs pollution, where sediments are most likely the final sink. MPs ingestion has been reported in a wide variety of marine species, still the ecotoxicological effectson marine ecosystems is poorly understood. The benthic bivalve Scrobicularia plana was selected for its capacity to filter pollutants and the role it plays in the functioning of estuarine communities. For this study, organisms of two size classes (big and small) were exposed to relevant environmental virgin polyethylene (PE) microparticles (75-90 µm) and also higher concentrations (C1 = 30 MPs L-1, C2 = 90 MPs L-1, C3 = 150 MPs L-1, and C4 = 450 MPs L-1) to assess the ecotoxicological and biochemical effects of this pollutant. A battery of biomarkers was selected to determine the ecotoxicological effects of PE microparticles and to assess the potential use of enzymatic
biomarkers for PE bioassays, including the activity of antioxidant enzymes (glutathione peroxidase (tGPx), glutathione reductase (GRed), glutathione-S-transferase (GST), Catalase (CAT) and Superoxide dismutase (SOD)) and the lipid peroxidation
occurrence (TBARS). PE microparticles did not show lethal effects for S. plana. However, alterations occurred in the activities of the antioxidant enzymes and lipid peroxidation. Clams of both size classes showed dose-dependent responses at the highest concentrations (450 MPs L-1), significantly increasing the activities of GST and GRed. A trend is observed between the
antioxidant enzymes CAT, SOD, and tGPx for both size classes. The results indicate the suitability of S. plana as an indicator for the presence of virgin PE microparticles in aquatic environments. Furthermore, this work reports that GST activity increases significantly under the highest concentration of MPs (450 MPs L-1), suggesting to be the best enzymatic biomarker to assess the presence of PE microparticles
1.01.P-Mo005 Toxicity of Cigarette Butts and Their Associated Chemicals on the Freshwater Invertebrate Chironomus riparius
Ms. Francesca Molinari1, Marion Blanchard2, Therese Nitschke3, Bethanie Carney Almroth4 and Agathe Bour3, (1)Department of Biology and Environmental Sciences, Univerity of Gothenburg, Göteborg, Sweden, (2)Ecole Nationale Supérieure
Agronomique de Toulouse (ENSAT), France, (3)University of Gothenburg, Sweden, (4)Goteborg University, Sweden
Cigarette butts are one of the most commonly found litter items in the global environment. Their distribution spans from streets to beaches, rivers and oceans. Once ignited, the smoke produced by the tobacco mixture contains many harmful substances such as nicotine, PAHs, formaldehyde, hydrogen cyanide, lead and arsenic. These substances are partially trapped into the cigarette filter, which, once present in the environment, can release the chemicals into the environment. While the effects of smoking cigarettes are well understood, the effect of cigarette butts on the environment is still to be better researched. The aim of this study was to investigate the effects of cigarette butt leachates on the aquatic environment. For this purpose, we used the model organism Chironomus riparius (midge larvae), a widely spread and abundant organism in freshwater ecosystems. The invertebrate larvae are in direct interaction with both sediment and water, making them representative of both compartments, an interesting trait for the experiments we carried out. Cigarette butts were left to leach in containers containing sand and freshwater. The two matrices were then separated. Chironomid larvae were exposed either to contaminated sand or contaminated water, complemented with clean water or sand, respectively. This experimental design allowed us to study where the cigarette leachates are more prone to accumulate and therefore which organisms, benthic or pelagic, would be most at risk. We also followed a multi-endpoint approach, exposing the organisms at different life stages and for different amounts of time (from 7 days to 3 months), assessing endpoints such as growth, development and teratogenicity. Our results show strong evidence that leachates from cigarette butts impact C. riparius. More pronounced effects were observed when comparing the sediment phase to the water phase. This indicates that chemicals contained in cigarette butts can leach and enter both the sediment and water phases, but with a stronger affinity to sediment, potentially putting benthic organisms more at risk. Benthic organisms are essential for ecosystem
functioning. In cigarette polluted basins, a loss of benthic species might mean the collapse of an entire ecosystem. We suggest that more experiments be conducted on different species of benthic organisms in order to confirm that cigarette butts are particularly toxic to sediment dwelling organisms.
1.01.P-Mo006 Relevance of Mysid Shrimps Toxicity Tests for Risk Assessment in Europe
Guido Gonsior, Aquaculture, Ecotox, Marine Science, GG BioTech Design, Homberg (Ohm), Germany
Mysid shrimp (Americamysis bahia) shows high sensitivity to pollutants. This finding increases the impact of marine test systems on risk assessment. But it is not clear if this is also relevant for the European zone and if sensitivity of the mysid shrimps is comparable to common species in Europe. We present data of the sensitivity of the mysid shrimp Americamysis bahia, native to the east coast of the United States in comparison to species in the East and North Sea (e.g. Praunus flexuosus). These data might be useful to evaluate the relevance of mysid shrimps tests for the risk assessment in Europe.
1.01.P-Mo007 Biochemical, Physiological, and Reproductive Effects of Glyphosate (FAENA®) and 2,4-D (HERBIDEX®) on the Tropical Cladoceran Ceriodaphnia rigaudi
Alma Rodríguez-Miguel1, Armando Vega López2 and Felipe Fernando Martínez-Jerónimo2, (1)Instituto Politécnico Nacional- Escuela Nacional de Ciencias Biológicas, Mexico, (2)Escuela Nacional de Ciencias Biologicas, IPN, Mexico
Herbicides have contributed to increasing agricultural production, but the intensive usage of these chemicals can produce
significant damage in the environment, mainly through the affection of no target species. The herbicides glyphosate and 2,4-D are used worldwide for weed control; the first is a non-selective contact herbicide that inhibits the synthesis of aromatic amino acids needed for protein formation, whereas 2,4-D is an auxin hormone that inhibits the cellular division of meristematic tissue, causing the reduction of apical growth in plants. Aquatic organisms are exposed to herbicides reaching the water by leaching, running-off,
or adsorption to particulate matter. Daphnia magna is a Cladoceran frequently used in toxicity studies. However, it is important to consider tropical species such as Ceriodaphnia rigaudi, a zooplankter distributed in México. The objective of this study was to determine the toxic effects that the mixture of glyphosate- and 2,4-D-based commercial formulations produce in the reproduction, physiology, and biochemistry of C. rigaudi. LC50’s (48h) (25°C, 16:8h light: darkness) were determined to be 1.09 mg L-1 for glyphosate and 233 mg L-1 for 2,4-D. Then, a chronic toxic assay was carried out during a whole life cycle. The tested concentrations were 0.0625, 0.125, 0.25, and 0.5 Toxic Units (TU) of the mixture of FAENA® and HERBIDEX®. During the tests, samples of 30 neonates from each clutch were picked up to analyze effects. The concentration of proteins, carbohydrates, and lipids and the activity of the enzymes CAT, SOD, and GPx were determined for each clutch. The 0.5 UT mixture of herbicides significantly decreased longevity. There was a delay of one day at the age of first reproduction in 0.125 and 0.5 UT concentrations. The accumulated progeny and the number of clutches decreased significantly at 0.5 UT. Abortions increased as the concentration of the mixture did. The content of proteins increased in the highest concentrations (0.125, 0.25, and 0.5 UT).
Carbohydrates decreased up to 80% compared with the control, and lipids were significantly different in the treatment of 0.125 UT. The activity of the SOD was stimulated twice, but the activity of CAT and GPx was inhibited. The simultaneous presence of two herbicides in the water may lead to increased toxicity, causing more negative effects on aquatic organisms than that expected in single exposures.
1.01.P-Mo008 Finding the Optimal Conditions for Ecotoxic Experiments With the Larvae of the Terrestrial Land Crab Cardisoma guanhumi
Concepcion Rodriguez-Fourquet2, Concepcion Rodriguez-Fourquet2 and Ms. Marie Corujo2, (1)University of Puerto Rico at Bayamon, United States, (2)University of Puerto Rico at Bayamon, Puerto Rico
The land crab Cardisoma guanhumi, one of the keystone species on the island of Puerto Rico, has a complex life cycle composed of a terrestrial adult stage and a larval aquatic stage; composed of zoea and megalopa stages. During this aquatic phase,
C.guanhumi zoeas are affected by environmental factors such as food availability, temperature, and salinity, affecting the
metamorphosis during the larval stage. Given this, we established the goal of discovering the ideal conditions for the development of C. guanhumi larval stages and determining the set of conditions necessary for developing phthalates bioassays. Gravid C.
guanhumi females were captured in the wild and transported to the laboratory. Zoeas were removed from the females and immediately transferred to the experimental vials. The zoeas were exposed to three different temperatures, three different salinities, and two different food regimes. The zoeas were also subjected to several treatments such as various water quality, buffers prepared with various water quality, and different glassware cleaning conditions. The survival was recorded in each of the experiments. We found that the zoea exposed to 35 ppt and temperatures between 21°C and 27°C showed the highest survival.
The food regimes showed that an excess and constant supply of Artemia salina newly hatched nauplii showed a tendency to higher survival of zoeas. The exposure of C. guanhumi zoeas to different water quality conditions, to buffers prepared with different water quality conditions, and to different glassware cleaning regimes showed no survival difference among the treatments. We concluded that we had identified the optimal conditions for future toxicity experiments with phthalate.
1.01.P-Mo009 Investigation of Different Food Sources for a Chronic Laboratory Test With Cloeon dipterum Julius Alberti, RWTH University Aachen, Aachen, Germany
In routine toxicity testing or regulatory evaluation of chemicals, standard organisms that are easy to maintain are often used.
These organisms are not always sensitive enough to ensure a correct evaluation of the chemical. Besides the standard organisms, there are non-standard organisms like mayflies which are very sensitive to certain chemicals. In recent years, there has been a growing interest in keeping a mayfly species in the laboratory for toxicity testing. An important aspect for keeping mayflies is the food source. It has been shown that the development rate of the mayfly larvae is directly related to the quality of the offered feed.
To reproduce comparable results, the food source must provide enough energy for the larvae to develop well, and any mortality or developmental delay effect must be due to the substance being tested. In this work, different food sources for the mayfly species Cloeon dipterum were investigated regarding their practicability and quality. The tests were carried out with locally collected larvae of C. dipterum. For each test set 6 replicates with 5 larvae each were prepared. First two fish foods and periphyton cultivated on tiles were tested. Secondly, incubated poplar leaves as whole and as a powder were tested. Again, in parallel with periphyton. The experiments were stopped as soon as all larvae had emerged or had died. The development of the larvae was examined 3 times a week. In order to investigate an increase of mortality caused by the monitoring procedure, a further non- invasive test with periphyton tiles as food source was performed. Here, only the vitality was checked every 7 days. In the test with the artificial fish food 1 15% of the larvae emerged while, 85% of the larvae died. With the artificial fish food 2 only 55%
mortality was observed. The parallel periphyton test showed the lowest mortality (30%). In the second run, the mortality in the periphyton test was higher (80%). Feeding with leaves resulted a slower development of the larvae, resulting in an increased duration of the test. The mortality was 55% - 60% and first dead larvae were observed early in the test. In general, larvae fed with fish feed developed slower compared to the test with periphyton tiles. Also, larvae died earlier and in smaller stages. In the non- invasive test, 40% of the larvae died. The regular examination of the larvae does not seem to have a negative influence on the test.
Overall, the periphyton tiles turned out to be a promising feed for larvae of C. dipterum.
1.01.P-Mo010 Behavioural and Enzymic Activities Changes Under Chronic Exposure of Neurotoxic and Non-Neurotoxic Pharmaceutical in Benthic Invertebrate
Ms. Wing Sze Chan1, Henriette Selck2 and Martina Santobuono2, (1)Roskilde University, Roskilde, Sweden, (2)Roskilde University, Roskilde, Denmark
Among the emerging contaminants, pharmaceuticals are considered one of the most pertinent substances that may pose threats to
the aquatic ecosystems. Pharmaceuticals are designed to be directed at specific metabolic and molecular pathways in selected organisms. Thus, they are assumed to still be biologically active when entering the ecosystem and may therefore result in unpremeditated impacts on non-target organisms. Neurotoxic and non-neurotoxic pharmaceuticals have different mode of action and therefore cause different impacts on the organisms. In addition, environmental stressors, such as food availability and temperature, may interact with contaminant exposure to affect uptake and action of pharmaceuticals in organisms. An enhanced understanding of the mode of action of pharmaceuticals in aquatic organisms under different environmental stressors is crucial to assess the possible impacts at organism to population level.This project examines the chronic impacts of low concentrations of sediment-associated pharmaceuticals: Fenofibrate (lipid regulator), naproxen (nonsteroidal anti-inflammatory drug), fluoxetine and sertraline (antidepressants) to the deposit-feeding oligochaete, Tubifex tubifex, using non-conventional endpoints (i.e., burrowing, feeding activity) and enzymatic activity (e.g., AChE, GST) following chronic exposure under different environmental settings.The presentation will discuss the approach to construct Adverse Outcome Pathways (AOPs) for the selected
pharmaceuticals: 1) a study on the changes of enzyme activities in T. tubifex under chronic exposure of neurotoxic and non- neurotoxic pharmaceuticals, 2) the use of non-conventional endpoints to assess the impact of chronic pharmaceuticals exposure on organisms, 3) an understanding of the impacts of environmental stressors on the mode of action of the pharmaceuticals and the response of the organisms, and 4) construction of AOPs to link behavioural and enzymic changes under chronic exposure of pharmaceuticals in benthic invertebrates.
1.01.P-Mo011 Effect of Diesel Particulate Matter on Caenorhabditis elegans: Neuro-Behaviour and Neurodegeneration Dr. Nivedita Chatterjee, INL International Iberian Nanotechnology Laboratory, Vila Nova de Gaia, Portugal
This study aims to elucidate whether the Diesel particulate matter (DPM) cause alterations in neuro-behaviours in C.elegans.
Potential long term toxicity (7 days) and acute toxicity (24h) were assessed in the dose range of 0.1 to 100 µg/ml. We found significant alterations in stress response, locomotion behaviour at sub-lethal dose. In connection with neuro-behavioral (dat-1) analysis, we found that marked impairment in dopaminergic function. C.elegans is an excellent model for neurodegenerative disease, such as Alzheimer’s disease (AD), characterized by amyloid protein aggregation, otherwise soluble proteins. Our preliminary data evidenced that DPM cause amyloid protein aggregation in the model system (ongoing study). In summary, DPM exposure could impair neuro-behaviour and accelerate the ageing-related neurodegenerative disease in the model system.
1.01.P-Mo012 High Throughput Investigation of Amphipod Reproductive Behaviour
Bidemi Green-Ojo1, Lena Grinsted2, Matthew Parker2 and Alex Ford2, (1)University of Portsmouth, Portsmouth, United Kingdom, (2)University of Portsmouth, United Kingdom
Precopulatory pairing is a reproductive strategy in which a mature male holds a mature female until she moults and copulation occurs. The use of precopulatory pairing as a behavioural endpoint in ecotoxicology was first demonstrated in the 1970s where precopulatory pairing was used to determine mating success. This qualitative behavioural response can be measured as time of disruption of pairs, repairing time or post-dosing observation of pairs for reproductive success. Although this mechanism is sensitive, there has been a steady decline in its use in toxicity testing as it can be onerous. This study aims to provide an overview of past and current trends on the use of precopulatory pairing and discuss the optimisation of this technique using video tracking technology to evaluate the effects of plastic additives on the marine amphipod, Echinogammarus marinus. Pairs were separated and individuals were exposed to high concentrations (50 and 5000) µg/l of di-ethylhexyl phthalate (DEHP) and dibutyl phthalate (DBP). The repairing activity was recorded using Zantiks MPV unit for 15 mins. The repairing time and the percentage of animals paired in the control and the exposed group were analysed. This approach to toxicity testing is a sensitive, reliable and cost- effective means of detecting the early warning effects of harmful environmental pollutants.
1.01.P-Mo013 Sertraline and Fluoxetine Chronic Exposure - Possible Sublethal Effects on the Key Benthic Invertebrate Capitella teleta
Martina Santobuono1, Ms. Wing Sze Chan2 and Henriette Selck1, (1)Roskilde University, Roskilde, Denmark, (2)Roskilde University, Roskilde, Sweden
Sertraline and fluoxetine are two selective serotonin reuptake inhibitors commonly prescribed to treat depression, anxiety, and post traumatic disorders in humans. Because Wastewater Treatment Plants (WWTPs) are not equipped to efficiently remove pharmaceuticals from wastewater, antidepressants are widely detected in the environment in the range of ng L-1 and µg L-1. These low concentrations might pose a risk to the resident species, especially in a long-term exposure. Since hydrophobic compounds easily sorb to organic matter, they tend to concentrate in sediments. Sertraline and fluoxetine are within these lipophilic chemicals, thus their presence in the water column could be less relevant than in sediment. However, not many studies have focused on sediment exposure of pharmaceuticals. Polychaetes are fundamental organisms for the geochemistry of benthic environments, and among them, Capitella teleta is a key species in organically enriched habitats, where it can reach densities of more than 100.000 individual/m2. We tested the impact of sediment-associated fluoxetine and sertraline on conventional (e.g., mortality) and non- conventional (e.g., burrowing- and feeding behavior) endpoints. Since antidepressants are neurotoxic chemicals, we also assessed acetylcholinesterase (AChE) enzymatic activity changes. The results will be discussed.
1.01.P-Mo014 Integument Colour Change: Tracking the Inhibition of Oppia Nitens Growth As a Sub-Lethal Indicator of Soil Toxicity
Kayode Jegede1, Hamzat Fajana2 and Steven Siciliano2, (1)Wageningen Environmental Research, Netherlands, (2)University of Saskatchewan, Canada
Growth is an important toxicity end-point in ecotoxicology but is rarely used in soil ecotoxicological studies. Here, we assessed
the growth change of Oppia nitens when exposed to reference and heavy metal toxicants. To assess mite growth, we developed an image analytic technique to measure colour spectrum change of the mite integument as a proxy for growth change. We linked the values of the red, green, blue, key-black and light colours of mites to different growth stages. Based on this concept, we assessed the growth change of mites exposed to cadmium, copper, zinc, lead, boric acid or phenanthrene at sublethal concentrations in LUFA 2.2 soil for 14 days. Sublethal effects were detected after 7 days of exposure. The growth of O. nitens was more sensitive than survival and reproduction when exposed to copper (EC50growth = 1360 mg/kg compared to EC50reproduction = 2896 mg/kg).
Mite growth sensitivity was within the same order of magnitude as mite reproduction, when exposed to zinc (EC50growth = 1785;
EC50reproduction = 1562 mg/kg). At least 25% sublethal effects of boric acid and phenanthrene were detected in the mites but growth was not impacted when O. nitens were exposed to lead. Consistent with previous studies, cadmium was the most toxic metal to O.
nitens. The mite growth pattern was comparable to mite survival and reproduction from previous studies. Mite growth is a sensitive toxicity endpoint, ecologically relevant, fast, easy to detect, and can be assessed in a non-invasive fashion, thereby complementing existing O. nitens testing protocols.
1.01.P-Mo015 The Effects of Life Stage on the Sensitivity of Folsomia candida to Cyproconazole and Teflubenzuron Saúl Fernandes1, Kees Buskermolen2, Diana Ilyaskina3, Matty Berg2 and Cornelis A.M. (Kees) van Gestel1, (1)Vrije Universiteit Amsterdam, Netherlands, (2)Vrije University Amsterdam, Netherlands, (3)Environment & Health, Vrije Universiteit Amsterdam, Amsterdam, Netherlands
To halt the loss of ecosystem services provided by terrestrial ecosystems, it is increasingly important to understand the effects of chemical stressors on the role of service providing units (SPU). Collembola are soil micro-arthropods with relevant roles in organic matter decomposition and nutrient cycling. For this reason, the registration of active ingredients used in plant protection products requires the assessment of their long-term effects on species of Collembola, such as Folsomia candida. The guidelines provided by the Organisation for Economic Cooperation and Development for such tests recommend the use of juvenile age- synchronized animals with 10 to 12 days old. In several studies these guidelines are modified by, e.g., using older animals at the beginning of the test, to compare the effects of an active ingredient with species of Collembola with slower life cycles. To determine the influence of the age of F. candida on the results of the toxicity test, we assessed the effect of two active ingredients on F. candida with an age difference of 10 days old. The fungicide cyproconazole (CYP) and the insecticide Teflubenzuron (TEF) were selected to clarify if the differences in sensitivity caused by the age of animals are observed for chemicals with distinct modes of action. After exposure to the chemicals, the dose-response curves obtained for mortality and reproduction of 10-12 or 20-22 days old springtails were compared. For CYP, the EC50 (122 mg kg-1soildw) for the effects on reproduction was significantly lower for the older compared to the juvenile springtails (EC50 of 241 mg kg-1soildw). For TEF, the LC50 for the younger
springtails was significantly lower (0.51 mg kg-1soildw) compared to that for the older animals (0.91 mg kg-1soildw). Even though the reproduction of both life stages was significantly inhibited by TEF, no significant difference was observed between different ages. These results show that the life stage of F. candida at the beginning of the toxicity tests can significantly influence the results obtained. Additionally, the influence of animal age on the test outcome was partially dependent on the active substance tested. This life-stage dependent effect should be considered in future studies to understand how pesticides can influence the role of SPU.
1.01.P-Mo016 Untargeted Lipidomics to Assess the Response of the Non-Targeted Species Folsomia candida to Sub-Lethal Concentrations of Teflubenzuron
Diana Ilyaskina1, Marissa Kateva2, Saúl Fernandes2, Marja Lamoree3 and Pim Leonards2, (1)Environment & Health, Vrije Universiteit Amsterdam, Amsterdam, Netherlands, (2)Vrije Universiteit Amsterdam, Netherlands, (3)VU University, Department Environment & Health, Netherlands
Increased rates of pesticide production brought serious concerns regarding their impact on non-targeted organisms in the
environment. Soil organisms as Collembola (or springtails) widely contribute to soil fertility and the nutrient cycle in ecosystems.
Therefore, springtails are an important model for ecotoxicological risk assessment (ERA). Classical approaches as reproduction, growth, and mortality are the main test systems used in ERA. Molecular ""omics"" studies have become an important part of ERA to study the mode of action of pesticides for non-target species. Untargeted screening of molecular changes can provide a more detailed picture of the sub-lethal effects of chemicals present in low concentrations in the environment. In this work, untargeted lipidomics was applied to study the potential toxicity effects of Teflubenzuron on the springtails Folsomia candida. Springtails were exposed (7 days) to three environmentally relevant concentrations (0.0056, 0.0139, and 0.0348 mg/kg soil) of
Teflubenzuron. Teflubenzuron showed a wide range of significant alterations in the lipid profile. Mainly phospholipids (PC and PE), triglycerols (TG), and fatty acids (FA) were affected. These lipids are primarily involved in membrane synthesis, lipid storage and energy metabolism. Teflubenzuron exposure showed an increase of oxidised forms of TG and FA in springtails which can be indicators of oxidative stress. Moreover, the increase of oxidised polyunsaturated FA (PUFA), shown on all levels of exposure, may cause severe problems since PUFA play a vital role in organisms' development. The decrease of TG can probably be linked to stored lipids which were used as an additional energy source to cope with stress. The decrease in TG in combination with the upregulation of monounsaturated FA and sterol esters indicates the balancing of lipid storage in the body. Dysregulation of PC and PE lipids, associated with the endoplasmic reticulum (ER) and mitochondria, can indicate ER stress. ER is closely connected with lipid droplets synthesis and regulation. Alteration of PE can indicate the disruption of the mitochondria membrane and could further lead to oxidative stress and potential malfunction of glucose metabolism. Hence, a combined study of lipidomics and classical ERA approaches can contribute to a better understanding of the toxicity of Teflubenzuron from the molecular to the population level.
