Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

MRNA-Seq of daphnia magna juveniles exposed to different sub-lethal concentrations of Fipronil in a modified acute immobilisation test below against untreated control groups

ABSTRACT: The aim of this mRNA expression profiling experiment was to screen for ecotoxicogenomic fingerprints in juvenile daphnids (daphnia magna) as aquatic vertebrate non-target model exposed to sub lethal concentrations of Fipronil. Fipronil is a widely used insecticide applied for crop protection against field pests and Ectoparasites. The Insecticide Resistance Action Committee (IRAC) classified Fipronil after its mode of action (MoA) in the target organism as a GABA-gated chloride channel blocker (Group 2). The goal is to identify toxicogenomic profiles with predictive character and identify potential molecular key events (KE) explaining upstream adverse effects. This will provide useful information to refine and improve existing adverse outcome pathways (AOP). Furthermore, integrating the obtained profiles for this and other tested chemicals in a collective database will enable us in the future to derive predictions about the ecotoxicological hazard for chemcials with unknown apical effects, based on similarly altered transcriptomic and proteomic profiles. In a modified version of the acute immobilisation test (OECD 202), 50 juvenile Daphnids were exposed to two sub lethal concentrations of Fipronil for 48 hours under static conditions. Each test comprised of a low exposure (LE), high exposure (HE) and a negative control (NC) group and was performed in triplicates. At 48 hours after introducing the daphnids into the test solutions, RNA and protein was extracted from living daphnids with NucleoSpin RNA/Protein kit (Macherey-Nagel). RNA quality was assessed with a 2100 Bioanalyzer system (Agilent) before messenger RNA was purified (PolyA selection with TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina HiSeq 4000 System (Illumina) in 50 bp single read mode, producing roughly 30 million reads per sample. Adapter sequences were removed with trimmomatic and sequences were aligned to the D. magna reference genome (daphmag2.4) using STAR. Counting of feature mapped reads was performed through featureCounts. Library gene count tables were then merged to a single count matrix as input for count normalization and differential gene expression analysis with DESeq2.

INSTRUMENT(S): Illumina HiSeq 4000

ORGANISM(S): Daphnia magna

SUBMITTER: Julia Pfaff

PROVIDER: E-MTAB-9829 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Toxicogenomic differentiation of functional responses to fipronil and imidacloprid in Daphnia magna.

Pfaff Julia J Reinwald Hannes H Ayobahan Steve U SU Alvincz Julia J Göckener Bernd B Shomroni Orr O Salinas Gabriela G Düring Rolf-Alexander RA Schäfers Christoph C Eilebrecht Sebastian S

Aquatic toxicology (Amsterdam, Netherlands) 20210727

Active substances of pesticides, biocides or pharmaceuticals can induce adverse side effects in the aquatic ecosystem, necessitating environmental hazard and risk assessment prior to substance registration. The freshwater crustacean Daphnia magna is a model organism for acute and chronic toxicity assessment representing aquatic invertebrates. However, standardized tests involving daphnia are restricted to the endpoints immobility and reproduction and thus provide only limited insights into the u ...[more]

PMID: 34340001

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Project description:The aim of this mRNA expression profiling experiment was to screen for ecotoxicogenomic fingerprints in juvenile daphnids (daphnia magna) as aquatic vertebrate non-target model exposed to sub lethal concentrations of Imidacloprid. Imidacloprid is is a popular insecticide widely applied in agriculture and by veterinarians against ectoparasites.The Insecticide Resistance Action Committee (IRAC) classified Imidacloprid after its mode of Action (MoA) in the target organism as a nicotinic acetylcholine receptor (nAChR) competitive modulator (Group 4). The goal is to identify toxicogenomic profiles with predictive character and identify potential molecular key events (KE) explaining upstream adverse effects. This will provide useful information to refine and improve existing adverse outcome pathways (AOP). Furthermore, integrating the obtained profiles for this and other tested chemicals in a collective database will enable us in the future to derive predictions about the ecotoxicological hazard for chemcials with unknown apical effects, based on similarly altered transcriptomic and proteomic profiles. In a modified version of the acute immobilisation test (OECD 202), 50 juvenile Daphnids were exposed to two sub lethal concentrations of Imidacloprid for 48 hours under static conditions. Each test comprised of a low exposure (LE), high exposure (HE) and a negative control (NC) group and was performed in triplicates. At 48 hours after introducing the daphnids into the test solutions, RNA and protein was extracted from living daphnids with NucleoSpin RNA/Protein kit (Macherey-Nagel). RNA quality was assessed with a 2100 Bioanalyzer system (Agilent) before messenger RNA was purified (PolyA selection with TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina HiSeq 4000 System (Illumina) in 50 bp single read mode, producing roughly 30 million reads per sample. Adapter sequences were removed with trimmomatic and sequences were aligned to the D. magna reference genome (daphmag2.4) using STAR. Counting of feature mapped reads was performed through featureCounts. Library gene count tables were then merged to a single count matrix as input for count normalization and differential gene expression analysis with DESeq2.

Project description:The aim of this mRNA expression profiling experiment was to screen for ecotoxicogenomic fingerprints in zebrafish (Danio rerio) embryos as aquatic vertebrate non-target model exposed to sub lethal concentrations of the heavily used organophosphate insecticide Fipronil (CAS 2921-88-2). The Insecticide Resistance Action Committee (IRAC) classified Fipronil after its mode of action (MoA) in the target organism as an acetylcholinesterase (AChE) inhibitor (Group 1B). The goal is to identify toxicogenomic profiles with predictive character and potential molecular key events (KE) explaining upstream adverse effects in aquatic non-target organisms. This will provide useful information to refine and improve existing adverse outcome pathways (AOP). Furthermore, integrating the obtained profiles for this and other tested chemicals in a collective database will enable us in the future to derive predictions about the ecotoxicological hazard for chemcials with unknown apical effects, based on similarly altered transcriptomic and proteomic profiles. In a modified version of the zebrafish embryo toxicity test (OECD 236), 15 fertilized eggs were exposed to two different sub lethal concentrations of Fipronil for 96 hours under semi-static conditions. Each test comprised of a low exposure (LE, 0.075 mg/L), high exposure (HE, 0.3 mg/L) and negative control (NC) group and was performed in triplicates. At 96 hours post fertilization (hpf), 10 larvae were randomly picked for each sample and pooled for RNA and protein extraction with NucleoSpin RNA/Protein kit (Macherey-Nagel). RNA quality was assessed with a 2100 Bioanalyzer system (Agilent) before coding RNA was purified (PolyA selection with TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina HiSeq 4000 System (Illumina) in 50 bp single read mode, producing roughly 30 million reads per sample. Adapter sequences were removed with trimmomatic and sequences were aligned to the D.rerio reference genome GRCz11 with STAR. Counting of feature mapped reads was performed through featureCounts. Library gene count tables were then merged to a single count matrix as input for differential gene expression analysis with DESeq2.

Project description:Testing for toxicity in a number of aquatic organisms is necessary for risk assessments of substances. Yet, aquatic larvae of the so-called EPT (Ephemeroptera, Plecoptera, Trichoptera) taxa are regularly exposed to several environmental contaminants and have been demonstrated to be extremely vulnerable to a variety of environmental pollutants. These results show that existing toxicity testing can result in an underestimating of the risk for EPT taxa and that more toxicity data using EPT taxonomic representatives are needed. Unfortunately, there is a dearth of standardized test techniques and scant published data, particularly for European EPT species. Our study's objective was to create a testing framework for a variety of endpoints in the mayfly Cloeon dipterum. Due to its high prevalence in local waterbodies and its brief lifecycle of a few weeks in the right environmental conditions, C. dipterum was selected as the test organism. To this end, two chronic toxicity tests with semi-static test design and two media renewal per week were performed. Small larvae in the stage L3 based on the wing pad development described by Cianciara (1976) were used in the tests. Four replicates per test concentration and control containing five individuals per replicate were installed. The emergence was determined daily on working days. The tests were conducted at a temperature of 20 °C (± 1 °C) and the illumination was < 1 µE m2s 1 with a light and dark rhythm of 16:8 h. The physico-chemical parameters pH, oxygen concentration, and oxygen saturation were measured using the multiparameter device WTW Multi 1970i at test start, throughout each media renewal, and at test end. Continuous temperature measurements were taken, and weekly lighting assessments were made. Fipronil was administered to C. dipterum larvae in the long-term exposure experiment at nominal concentrations ranging from 0.038 to 0.60 g/L. This test was conducted for 38 days until all larvae had emerged. In the short-term exposure experiment, C. dipterum larvae were exposed to Fipronil in nominal concentrations between 0.038 and 0.30 µg/L for a test duration of seven days. At test end, the larvae were sampled for transcriptome analysis.

Project description:The aim of this mRNA expression profiling experiment was to screen for ecotoxicogenomic fingerprints in zebrafish (Danio rerio) embryos as aquatic vertebrate non-target model exposed to sub lethal concentrations of Abamectin (CAS 71751-41-2). Abamectin is a heavily used insecticide applied for crop protection against sucking insects (i.e. Acari). The Insecticide Resistance Action Committee (IRAC) classified Abamectin after its mode of action (MoA) in the target organism as a Glutamate-gated chloride channel (GluCl) allosteric modulator (Group 6). In vertebrates, GluCl do not exist, but they are closely related to vertebrate glycine receptors (Wolstenholme 2012). The goal is to identify toxicogenomic profiles with predictive character and potential molecular key events (KE) explaining upstream adverse effects in aquatic non-target organisms. This will provide useful information to refine and improve existing adverse outcome pathways (AOP). Furthermore, integrating the obtained profiles for this and other tested chemicals in a collective database will enable us in the future to derive predictions about the ecotoxicological hazard for chemcials with unknown apical effects, based on similarly altered transcriptomic and proteomic profiles. In a modified version of the zebrafish embryo toxicity test (OECD 236), 15 fertilized eggs were exposed to two different sub lethal concentrations of Abamectin for 96 hours under semi-static conditions. Each test comprised of a low exposure (LE, 0.11 mg/L), mid exposure (ME, 0.22 mg/L), high exposure (HE, 0.44 mg/L) and negative control (NC) group and was performed in triplicates. At 96 hours post fertilization (hpf), 10 larvae were randomly picked for each sample and pooled for RNA and protein extraction with NucleoSpin RNA/Protein kit (Macherey-Nagel). RNA quality was assessed with a 2100 Bioanalyzer system (Agilent) before coding RNA was purified (PolyA selection with TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina HiSeq 4000 System (Illumina) in 50 bp single read mode, producing roughly 30 million reads per sample. Adapter sequences were removed with trimmomatic and sequences were aligned to the D.rerio reference genome GRCz11 with STAR. Counting of feature mapped reads was performed through featureCounts. Library gene count tables were then merged to a single count matrix as input for differential gene expression analysis with DESeq2.

Project description:The aim of this mRNA expression profiling experiment was to screen for ecotoxicogenomic fingerprints in zebrafish (Danio rerio) embryos (96 hpf) as aquatic vertebrate non-target model exposed to sub lethal concentrations of the carbamate insecticide Carbaryl (CAS 63-25-2). The Insecticide Resistance Action Committee (IRAC) classified Carbaryl after its mode of action (MoA) in the target organism as an acetylcholinesterase (AChE) inhibitor (Group 1A). The goal is to identify toxicogenomic profiles with predictive character and potential molecular key events (KE) explaining upstream adverse effects in aquatic non-target organisms. This will provide useful information to refine and improve existing adverse outcome pathways (AOP). Furthermore, integrating the obtained profiles for this and other tested chemicals in a collective database will enable us in the future to derive predictions about the ecotoxicological hazard for chemcials with unknown apical effects, based on similarly altered transcriptomic and proteomic profiles. In a modified version of the zebrafish embryo toxicity test (OECD 236), 15 fertilized eggs were exposed to two different sub lethal concentrations of Carbaryl for 96 hours under semi-static conditions. Each test comprised of a low exposure (LE, 275 µg/L), high exposure (HE, 1100 µg/L) and negative control (NC) group and was performed in triplicates. At 96 hours post fertilization (hpf), 10 larvae were randomly picked for each sample and pooled for RNA and protein extraction with NucleoSpin RNA/Protein kit (Macherey-Nagel). RNA quality was assessed with a 2100 Bioanalyzer system (Agilent) before coding RNA was purified (PolyA selection with TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina HiSeq 4000 System (Illumina) in 50 bp single read mode, producing roughly 30 million reads per sample. Adapter sequences were removed with trimmomatic and sequences were aligned to the D.rerio reference genome GRCz11 with STAR. Counting of feature mapped reads was performed through featureCounts. Library gene count tables were then merged to a single count matrix as input for differential gene expression analysis with DESeq2.

Project description:The aim of this mRNA expression profiling experiment was to screen for ecotoxicogenomic fingerprints in zebrafish (Danio rerio) embryos as aquatic vertebrate non-target model exposed to sub lethal concentrations of synthetic organochloride insecticide Methoxychlor (CAS 72-43-5). It is structurally highly similar to its precursor molecule DDT (Dichlorodiphenyltrichloroethane). Like its precursor, today Methoxychlor is banned from the use as pesticide in the United States and the European Union due to its acute toxicity, high bioaccumulation potential and endocrine disruption activity. The Insecticide Resistance Action Committee (IRAC) classified Methoxychlor after its mode of action (MoA) in the target organism as a sodium channel modulator (Group 3B). Our goal is to identify toxicogenomic profiles with predictive character and potential molecular key events (KE) explaining upstream adverse effects in aquatic non-target organisms for this substance of particularly high environmental concern. This will provide useful information to refine and improve existing adverse outcome pathways (AOP). Furthermore, integrating the obtained profiles for this and other tested chemicals in a collective database will enable us in the future to derive predictions about the ecotoxicological hazard for chemcials with unknown apical effects, based on similarly altered transcriptomic and proteomic profiles. In a modified version of the zebrafish embryo toxicity test (OECD 236), 15 fertilized eggs were exposed to two different sub lethal concentrations of Methoxychlor for 96 hours under semi-static conditions. Each test comprised of a low exposure (LE, 20 µg/L), mid exposure (ME, 60 µg/L), high exposure (HE, 180 µg/L) and negative control (NC) group and was performed in triplicates. At 96 hours post fertilization (hpf), 10 larvae were randomly picked for each sample and pooled for RNA and protein extraction with NucleoSpin RNA/Protein kit (Macherey-Nagel). RNA quality was assessed with a 2100 Bioanalyzer system (Agilent) before coding RNA was purified (PolyA selection with TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina HiSeq 4000 System (Illumina) in 50 bp single read mode, producing roughly 30 million reads per sample. Adapter sequences were removed with trimmomatic and sequences were aligned to the D.rerio reference genome GRCz11 with STAR. Counting of feature mapped reads was performed through featureCounts. Library gene count tables were then merged to a single count matrix as input for differential gene expression analysis with DESeq2.

Project description:The aim of this mRNA expression profiling experiment was to screen for ecotoxicogenomic fingerprints in zebrafish (Danio rerio) embryos as aquatic vertebrate non-target model exposed to sub lethal concentrations of the heavily used neonicotinoid insecticide Imidacloprid (CAS 138261-41-3). The Insecticide Resistance Action Committee (IRAC) classified Imidacloprid after its mode of action (MoA) in the target organism as a nicotinic acetylcholine receptor (nAChR) competitive modulator (Group 4A) The goal is to identify toxicogenomic profiles with predictive character and potential molecular key events (KE) explaining upstream adverse effects in aquatic non-target organisms. This will provide useful information to refine and improve existing adverse outcome pathways (AOP). Furthermore, integrating the obtained profiles for this and other tested chemicals in a collective database will enable us in the future to derive predictions about the ecotoxicological hazard for chemcials with unknown apical effects, based on similarly altered transcriptomic and proteomic profiles. In a modified version of the zebrafish embryo toxicity test (OECD 236), 15 fertilized eggs were exposed to two different sub lethal concentrations of Imidacloprid for 96 hours under semi-static conditions. Each test comprised of a low exposure (LE, 0.015 mg/L), medium exposure (ME, 0.03 mg/L), high exposure (HE, 0.06 mg/L) and negative control (NC) group and was performed in triplicates. At 96 hours post fertilization (hpf), 10 larvae were randomly picked for each sample and pooled for RNA and protein extraction with NucleoSpin RNA/Protein kit (Macherey-Nagel). RNA quality was assessed with a 2100 Bioanalyzer system (Agilent) before coding RNA was purified (PolyA selection with TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina HiSeq 4000 System (Illumina) in 50 bp single read mode, producing roughly 30 million reads per sample. Adapter sequences were removed with trimmomatic and sequences were aligned to the D.rerio reference genome GRCz11 with STAR. Counting of feature mapped reads was performed through featureCounts. Library gene count tables were then merged to a single count matrix as input for data normalization and differential gene expression analysis with DESeq2.

Project description:The aim of this mRNA expression profiling experiment was to screen for ecotoxicogenomic fingerprints in zebrafish (Danio rerio) embryos as aquatic vertebrate non-target model exposed to sub lethal concentrations of the heavily used organophosphate insecticide Chlorpyrifos (CAS 2921-88-2). The Insecticide Resistance Action Committee (IRAC) classified Chlorpyrifos after its mode of action (MoA) in the target organism as an acetylcholinesterase (AChE) inhibitor (Group 1B). The goal is to identify toxicogenomic profiles with predictive character and potential molecular key events (KE) explaining upstream adverse effects in aquatic non-target organisms. This will provide useful information to refine and improve existing adverse outcome pathways (AOP). Furthermore, integrating the obtained profiles for this and other tested chemicals in a collective database will enable us in the future to derive predictions about the ecotoxicological hazard for chemcials with unknown apical effects, based on similarly altered transcriptomic and proteomic profiles. In a modified version of the zebrafish embryo toxicity test (OECD 236), 15 fertilized eggs were exposed to two different sub lethal concentrations of 6PTU for 96 hours under semi-static conditions. Each test comprised of a low exposure (LE, 0.75 mg/L), high exposure (HE, 3 mg/L) and negative control (NC) group and was performed in triplicates. At 96 hours post fertilization (hpf), 10 larvae were randomly picked for each sample and pooled for RNA and protein extraction with NucleoSpin RNA/Protein kit (Macherey-Nagel). RNA quality was assessed with a 2100 Bioanalyzer system (Agilent) before coding RNA was purified (PolyA selection with TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina HiSeq 4000 System (Illumina) in 50 bp single read mode, producing roughly 30 million reads per sample. Adapter sequences were removed with trimmomatic and sequences were aligned to the D.rerio reference genome GRCz11 with STAR. Counting of feature mapped reads was performed through featureCounts. Library gene count tables were then merged to a single count matrix as input for data normalization and differential gene expression analysis with DESeq2.

Project description:The aim of this experiment was to develop an approach for the assessment of immunotoxicity in zebrafish embryos (Danio rerio) using transcriptomic profiling. Clobetasol propionate, a highly potent synthetic corticosteroid, which is known to have immunosuppressive effects on vertebrates including zebrafish, was used for continuous exposure from fertilization until extraction. As immunotoxicity can only be comprehensively assessed when the effect is observed during an acute infection, the embryos were injected with a mixture of different pathogen-associated molecular patterns (PAMPs) consisting of Pam3CSK4 (synthetic lipopeptide), poly(I:C) (synthetic viral dsRNA) and flagellin (isolated from Bacillus subtilis). Functional gene expression analysis from samples obtained by different kinds and combinations of treatments in relation to appropriate controls is used to identify pathways that are upregulated by artificial infection and suppressed by treatment with clobetasol propionate. In a modified version of the zebrafish embryo toxicity test (OECD 236), 20 fertilized eggs were exposed to a sub-lethal concentration (250 nM) of clobetasol propionate for 48 hours under semi-static conditions. The test comprised of three treatments with clobetasol propionate and two negative control (NC) groups and was performed in triplicates. At 46 hours post fertilization (hpf), the eggs were manually dechorinated. At 48 hpf, the embryos were immobilized by anaesthetization using 200 mg/mL tricaine. Using microinjection, 8 nL of the mixed PAMPs (0.33 µg/mL Pam3CSK4, 0.33 µg/mL poly(I:C) and 0.033 µg/mL flagellin) were injected directly into the embryos bloodstream. One of each exposure groups was injected with PAMPs, while another one was injected with ultrapure water to control for effects introduced by the injection process. One control group remained completely untreated. After injection, the embryos were further incubated for 3 hours before 10 embryos were randomly picked for each sample and pooled for RNA and protein extraction with NucleoSpin RNA/Protein kit (Macherey-Nagel). RNA quality was assessed with a 2100 Bioanalyzer system (Agilent) before coding RNA was purified (PolyA selection with TruSeq RNA Library Prep Kit v2) and sequenced on an Illumina HiSeq 4000 System (Illumina) in 50 bp single read mode. Adapter sequences were removed with trimmomatic and sequences were aligned to the D.rerio reference genome GRCz11 with STAR. Counting of feature mapped reads was performed through featureCounts. Library gene count tables were then merged to a single count matrix as input for differential gene expression analysis with DESeq2.

Dataset Information

MRNA-Seq of daphnia magna juveniles exposed to different sub-lethal concentrations of Fipronil in a modified acute immobilisation test below against untreated control groups

Publications

Toxicogenomic differentiation of functional responses to fipronil and imidacloprid in Daphnia magna.

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