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Ecotoxicology
Effects on individuals
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Significance/Relevance of acute tests
Effects on individuals – toxicity tests
• Acute toxicity values are no biological constants• Variations between laboratories and experimental
replicates • Differences among species and individuals at different life-
stages • Acute toxicity values are not sufficient for ecotoxicity-
evaluation • Results not transferable to chronic toxicity and no
inclusion of indirect (ecological) effects• Enhancing the significance by using several species, e.g.
SSDs
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Species Sensitivity Distributions (SSDs): e.g. CPF
Effects on individuals – toxicity tests Maltby et al. 2002
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Lecture contents
• IntroductionPhenomenons and principles
• Exposure (Environmental chemistry)Chemicals in the environment Classes of chemicals Chemical properties Processes
• Effects on individualsDose-response-relationships Acute and chronic toxicity Toxicity testsSubcellular tests, BiomarkerCombined effects
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Effects on individuals
Toxicity tests (contd)
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Overview of aquatic toxicity tests
Effects on individuals – toxicity tests Fent 2003
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Aquatic tests - Algae
Effects on individuals – toxicity tests
72-h EC50: inhibition of cell proliferation and photosynthesis
Fent 2003
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Aquatic tests - invertebrates
Effects on individuals – toxicity tests
Daphnia: Acute (48 h) swimming capacity, 21-days repro testChironomus: Acute (96 h) mortality, 28-days reproduction test
(sediment and water exposure)
Daphnia magna EUDaphnia pulex USChironomus riparius EUChironomus tentans US
Fent 2003
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Aquatic tests - fish
Effects on individuals – toxicity tests
Fish: - acute (96 h) mortality, - chronic 14-28 days mortality,-ELS, FFLC (fertility, reproduction),- in-vitro cell cultures (genotoxicity)
TroutFathead minnowBluegillZebrafishMedakaCarp
Fent 2003
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Comparison ELS and FFLC: NOECs for zebrafish
Effects on individuals – toxicity tests Fent 2003
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Effects on individuals
Subcellular tests, Biomarkers
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Molecular effects on cells
• Adverse effects on cellular processesa. Disturbance of the Ca 2+ - and Ion-metabolismIncrease of Ca-conc.: Activation of degradation enzymes, destruction
of the cytosceleton, membranes and DNA b. Inhibition of the energy synthesis (ATP, photosynthesis)c. Disturbance/inhibition of enzymatical processesGlutathione to protect Proteinthiole against binding of pollutantsd. Disturbance of the growth regulation
• Effects on cellular structuresInteraction with membrane structuresChange or destruction of membranesInfluence on organelles: mitochondria, lysosomesInfluence on cell nucleus: genotoxicity, cancerogenity
Effects on individuals - Subcellular tests, Biomarkers
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Expl: Molecular effects of organotin-compounds
Effects on individuals - Subcellular tests, Biomarkers Fent 2003
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Effects on structure and function of cell membranes
Effects on individuals - Subcellular tests, Biomarkers Fent 2003
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Permanent cell cultures: Permanent Hepatoma cell line
Germany, 1991: ca. 200 000 fish used in toxicity testing
Effects on individuals - Subcellular tests, Biomarkers Fent 2003
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Fish cell test for cytotoxicity
Effects on individuals - Subcellular tests, Biomarkers Fent 2003
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In vitro Neutralred-Assay
Effects on individuals - Subcellular tests, Biomarkers Fent 2003
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Biomarkers - Summary
• (Subcellular) biologic reaction to environmental chemicals Receptor bindingBiochemical response: Induction of detoxification enzymes, inhibition of
enzymesPhysiological response: avian eggshell-thinning, imposex of fish embryos
• Advantages and disadvantages of subcellular tests or biomarkers?• Advantages
SensitivityDetection of exposure and assessment of mechanisms of toxicityEthical aspects concerning vertebratesScreening tests
• DisadvantagesLack of knowledge on the whole organism: metabolism Chronic effects not well representedResults with little relevance for populations/communities/ecosystems
Effects on individuals - Subcellular tests, Biomarkers
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Effects on individuals
Combined effects
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Combined effects: overview
Effects on individuals – combined effects
• Two or more substances coexistant: „Mixture toxicity“• Pollutant and another abiotic stressor
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Mixture toxicity: Additive effects, Synergism, Antagonism
Effects on individuals – combined effects Moriarty 1988
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Calculating mixture toxicity (assumption: additive effect)
Effects on individuals – combined effects Hund & Kördel 1996
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Prediciting the mixture toxicity of an increasing no. of substances?
Effects on individuals – combined effects
Funnel Hypothesis
Warne & Hawker 1995
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Funnel Hypothesis: real data
Effects on individuals – combined effects Warne & Hawker 1995
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Mixture toxicity: substances with similar MoA
Effects on individuals – combined effects
Concentration addition
Grimme et al. 1998
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Mixture toxicity: substances with differing MoA
Effects on individuals – combined effects
Response addition
Grimme et al. 1998
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Fluoranthene and UV: Effects on frog embryos
Effects on individuals – combined effects
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Heavy metal contamination and UV in the Antarctic
Effects on individuals – combined effects
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Heavy metal contamination and UV in the Antarctic
Effects on individuals – combined effects
Survival of the amphipodParamorea walkeri
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Combined effects: overview
Effects on individuals – combined effects
• Two or more substances coexistentAdditivity, Antagonism, SynergismMixture toxicityFunnel Hypothesis
• Pollutant and another abiotic stressorUV-induced toxicity of PAHsHeavy metals and UV in the Antarctic
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• Examinations on different levels
• Important termsAcute, chronic, 96-h LC50, 48-h EC50, NOEC, LOEC, NOEAEC
• Linear and non-linear dose-response-relationships
• Acute and chronic toxicitySpecies differences up to factor 10000Acute (mortality) : chronic (sublethal) = 1 : 1 to 1 : 10000
Effects on individuals - summary
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• Species Sensitivity Distributions (SSDs)
• Ecotoxicological testsOECD (GLP), 91/414/EC, REACHAlgae, water-flea, fish as standardset
• Subcellular tests, BiomarkersAlternatives to fishtests?
• Combined effectsMixture toxicityChemical and abiotic stressor
Effects on individuals - summary
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Lecture contents (cont.)
• Effects on populations and communitiesMicro- and MesocosmsInteractions, indirect effectsIn situ systemsField studies: ecosystem-effects
• Risk assessment and risk managementAssessment schemesProbabilistic methodsAuthorization of chemicalsRisk mitigation strategies
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Organisational
• See you next week• Everybody to think about a question/statement related to
ecotox at the level of individuals for next Tuesday