1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif 1212570_28446780.jpg logo_mu_cerne.gif trojlogo.gif Introduction to Ecotoxicology Ludek Blaha + ecotox colleagues 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Global anthropogenic threats ? A safe operating space for humanity & the nine planetary boundaries Rockstrom et al. 2009 (Ecology and Society 14(2): 32; Nature 461, 472-475) 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •1996 - Chemicals in the environment •Do you believe that chemicals in products sold to consumers have been proven safe? • •Think again • • most chemicals in modern use have simply not been tested for their impacts on •human, even very basic effects. • •… what about the effects in nature, then ? OSF%20Hardback%20Large •How we stand 20 years later? 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif • 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif http://i.dailymail.co.uk/i/pix/2012/12/05/article-2243155-165C6117000005DC-311_634x522.jpg 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif • •Environmental pollution • • •Examples and ecological cosequences • 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Contamination of water - chemicals ? •Degradable „nontoxic“ •organic material + nutrients / fertilizers (N/P) •„Other“ chemicals 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Major anthropogenic threats – example: waters Impacts 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Loss of biodiversity – Major impacts 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Changes in biodiversity 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Changes in biodiversity •NATURE (2012) 482: 20 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Loss of biodiversity Major impacts •Impairment of ecosystem services –Unbalanced water cycles •Water scarcity •Draughts/floods –Impaired water quality •Drinking waters •Bathing waters •Toxicants in food chain –Shrinking of food supplies •Direct à lowering fish amounts •Indirect à crop yield 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Impacts on fish à decreased crop yields C:\Documents and Settings\Ludek\Dokumenty\katedra\vyuka\ObecEkotox\Nature-Ecosystem\nature03962-f1.jpg •NATURE (2005) 437: 880 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Impacts on biota à global effects •Mixing oceans •à cooling the atmosphere •[Nature 447, p.522, May 31, 2007] •Marine life supplies up to 50% of the mechanical energy required worldwide to mix waters from the surface to deeper cool layers •[Dewar, Marine Res 64:541 (2006)] •[Katija a Dabiri, Nature 460:624 (2009)] 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Ecotoxicology • •assessment o hazards •and risks of chemicals in ecosystems 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif • •Assessment of chemical hazards • •…to… • • Humans Other organisms • (TOXICOLOGY) (ECOtoxicology) • https://encrypted-tbn1.google.com/images?q=tbn:ANd9GcTE7qm7tLqavKV1VUwQpKDwC6XUksMmHXLKvoBzqHIKaqjv q_mACQ https://encrypted-tbn2.google.com/images?q=tbn:ANd9GcRorg12adBHNbxSdT8AiWMQmO7b7vltAdBZohD_pqfDehNn re7fkg https://encrypted-tbn1.google.com/images?q=tbn:ANd9GcQvHwZACb-TU516T7ZMDFe-QBU598rfeCvHryOWorunBunH oi4zpA 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Aim: to maintain the natural structure and function of ecosystems • •Definitions: §ecotoxicology is concerned with the toxic effects of chemical and physical agents on living organisms, especially on populations and communities within defined ecosystems; it includes the transfer pathways and their interactions with the environment §science of contaminants in the biosphere and their effect on constituents of the biosphere, including humans’ (Newman & Unger, 2002) §science that provides critical information on effects of toxic compounds on living organisms which SERVE various practical aims (environmental protection) ECOTOXICOLOGY by definition 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •CHEMICAL ENTERS THE ENVIRONMENT •Bioavailable fraction • •“EXPOSURE” •acute •chronic • •Toxicokinetics •biotransformation bioactivation excretion / sequestration •Target site •“EFFECT” •LEVELS, FATE, PROCESSES • •CHEMICAL ENTERS THE ORGANISM •biomonitoring • • • • 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Ecotoxic effects •Escher, B. I., Behra, R., Eggen, R. I. L., Fent, K. (1997), "Molecular mechanisms in ecotoxicology: an interplay between environmental chemistry and biology", Chimia, 51, 915-921. 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif • •Ecotoxicology - from molecules to ecosystems •… and backwards • • • 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •From ecosystems • •down the mechanisms •From mechanisms (molecules) • •up to effects and ecosystems •OR •? 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif 1962 •© Patuxent Wildlife Refuge, MA, USA •http://www2.ucsc.edu/scpbrg/ 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Bitman et al. Science 1970, 168(3931): 594 •Biochemistry bird carbonate dehydratase •In vivo: shell thinning •In situ: bioaccumulation -> bird population decline 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif 1) From molecules to individuals http://www.epa.gov/ncer/rfa/2010/comptox_001.jpg •MECHANISMS OF TOXICITY 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif 2) From molecules to individuals - AOPs •ADVERSE OUTCOME PATHWAYS Models for ecotoxicology and risk assessment. Toxicokinetics are "what the organism does with the chemical" and toxicodynamics are "what the chemical does to the organism". 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Ethinylestradiol •Binds to •ESTROGEN RECEPTOR https://www-pls.llnl.gov/data/assets/images/science_and_technology/life_sciences/cancer_diet/kulp1. jpg •Target genes - Proliferation/Apoptosis (sexual organs) - Synthesis of egg yolk (fish, amphibia) •Effects - Females: reproduction regulation - Males: feminization • (+ e.g. cancer promotion, development, • immunomodulation) AOP Example: ethinylestradiol 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Kidd, K.A. et al. 2007. Collapse of a fish population following exposure to a synthetic estrogen. Proceedings of the National Academy of Sciences 104(21):8897-8901 •Controls +Ethinylestradiol •5 ng/L (!) •7 years 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Molecular §Nonspecific effects §Hydrophobic interactions with phospholipid membranes (baseline = narcotic toxicity) §Direct reactivity: electrophilic compounds à nucleophilic organism (e.g. oxidation of PROTEINS, lipids (membranes), DNA …) §Specific effects §Activation of ER, AR and other „nuclear receptors“ §Inhibition of enzymes (e.g. CN- inhibits hemes in mitochondria/hemoglobin, insecticides …) § Effects at different levels - molecular https://encrypted-tbn2.google.com/images?q=tbn:ANd9GcSG1cHajHLtmVDjpZ78t4F7TH0Se4qLRsuMTInEY17Eupfv mEa4LQ 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Cellular §Effects on structure §Effects on metabolism (maintenance) §Effects on regulation àChanges in functions (e.g. Ethinylestradiol) àRepair, survival, growth àDeath (apoptosis or necrosis) àProliferation àDifferentiation Effects at different levels - cellular • • http://www.helmholtz-muenchen.de/uploads/pics/stem-cell-fate-options_09.gif 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Organism level – important in ecotoxicology (see Bioassays) §Effects on structure §Effects on metabolism (maintenance) §Effects on regulation § àChanges in functions (e.g. Ethinylestradiol) àRepair, survival, growth àDeath àProliferation = Reproduction Effects at different levels - ORGANISM • • •3 key apical endpoints •(reflected e.g. in regulations) 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Metabolism • •Control, •Interactions with environment •Defence against pathogens predators … • • • Defence against toxicants •Energy •hv •food •Losses •heat •faeces •Life •(maintenance) •Growth •to sexual maturity •Reproduction •Chemical stress 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Metabolism • •Control, •Interactions with environment •Defence against pathogens predators … • • • Defence against toxicants •Energy •hv •food •Losses •heat •faeces •Life •(maintenance) •Growth •to sexual maturity •Reproduction •Chemical stress •Chemical stress à energy re-allocation •à „insufficient“ resourses elsewhere 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Metabolism • •Control, •Interactions with environment •Defence against pathogens predators … • • • Defence against toxicants •Energy •hv •food •Losses •heat •faeces •Life •(maintenance) •Growth •to sexual maturity •Reproduction •Chemical stress •Chemical stress + ... another stress (food scarcity) 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Population –(… all the organisms that both belong to the same group or species (i.e. can sexually reproduce) and live in the same time within the same geographical area) §Effects on structure §elderly vs. young, males vs. females §Effects on maintenance & growth §Natality, mortality, reproduction fitness § Effects at different levels • • • • https://encrypted-tbn1.google.com/images?q=tbn:ANd9GcRy8TZLY8w9X0x--nqSFW4CqPPTUX3RdYucJtb5pGmKOWt- n2r_ 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Community & Ecosystem –(… a group of interacting living organisms sharing a populated environment) §Effects on structure §Loss of species, loss of biodiversity §Effects on functioning §(including „ecosystem functions“) § § § Effects at different levels • • • • http://www.combat-fishing.com/LowestReachesOZStreamSFW.JPG 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Ecotoxicology • •Science of doses / concentrations • •HAZARDS vs RISKS 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif • • • • • •RETROSPECTIVE •PROSPECTIVE •Bioassessment •Field assessment •Monitoring •Bioassessment •Field assessment •Monitoring •Lab studies •Lab studies •Simulated small •ecosystems •DISASTERS •PREDICTIONS for future •Time: NOW ! Ecotoxicology: problems and approaches •Most common in practice 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif 1 • •Bioassays •- single / multiple species •- acute / chronic effects •- standardized (practical) • vs. experimental (research) • •Simulation of the ecosystem •- major trophic levels • - producers • - consumers • - decomposers figure4 Eisenia_fetida scenedesmus_quadricauda_upr Testing ecotoxicity – basics 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif • • auto0 AcuteToxScheme_RM HaF-adult arenic~4 Daphnia Dania pict •Cu addition •Effect concentrations expressed in total/dissolved Cu •??? Safe concentrations ??? • • Ecotoxicology methods 1. - standardized assays 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif • •Expensive & time consuming (e.g. Pesticide testing) •Variable results (natural variability …) •Higher ecological relevancy • 1 1 figure4 Ecotoxicology – methods 2. Micro & Mesocosms 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •- complex issue (geology, climate, chemistry, biology ..) • Ecotoxicology mixes with Ecology •- comparing „contaminated“ with „control“ sites figure4 figure4 figure4 figure4 Ecotoxicology – methods 3. Field assessment / biomonitoring 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Notes on practical testing •Testing chemicals –Traditional / bioassays developed to assess individual chemicals –Advantages: Standardized approaches –Disadvantage: Limited ecological relevance •often acute tests only •„too standardized…“ (? Less representative ?) •does not assess/consider bioavailability •no consideration of mixture effects •no consideration of specific modes of action •no consideration of ecological situation – •Example: Acute (96h) fish toxicity assay with ethanol –No deaths (but fish are passive – slow swimming) à OK ? –Real life: easy prey à population decline • 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Notes on practical testing •Testing toxicity of natural contaminated matrices – –Rather new in ecotoxicology – many open challenges •Whole effluent toxicity testing (WET) •Contact soil toxicity assays – –More complex and more complicated •„cause-effects“ often not clear –Natural variability in matrices –Algal tests - nutrients (Nitrogen, Phosporus) >> Toxic compounds – 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Ecotoxicology in current practice •Most legislations on chemicals) (e.g. REACH, Pharmaceuticals, Pesticides) –have very simple (basic) requirements •EC50 from acute toxicity •Of 3 basic assays –Algae –Daphnia –Fish – –Ecotox database: –www.epa.gov/ecotox 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Ecotoxicology in current practice •How to extrapolate 3 (or few more) EC50 values to get legally binding safe concentration, which is protecting virtually all organisms? • • PNEC • (Predicted No Effect Concentration) • “value recommended by scientists” • • • EQS • (Environmental Quality Standard) • “value that occurs in legislation” • ? ? 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif • • • • Atmospheric Deposition • • Erosion & Runoff • Untreated discharges • • • • •Predicted Environmental •Concentration (PEC) • • bosmina FISH Laboratory (and field) studies Ecotoxicity tests sample56 lab2 faCTORY1 • •WWTP • j0173962 •effective concentrations •(PNEC) •Exposure •(resulting from load) •Effects •(what exposures cause effects ?) Cause – effect à Risk assessment 2 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Extrapolation approaches • •Data •Assessment factor •L(E)C50 short-term toxicity tests •NOEC for 1 long-term toxicity test •NOEC for additional long-term •toxicity tests of 2 trophic levels •NOEC for additional long-term •toxicity tests of 3 species of 3 •trophic levels •1000 •100 •50 •10 •Ecotoxicological data •Assessment / Extrapolation factors • •PNEC 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Extrapolation approaches • •Data •Assessment factor •L(E)C50 short-term toxicity tests •NOEC for 1 long-term toxicity test •NOEC for additional long-term •toxicity tests of 2 trophic levels •NOEC for additional long-term •toxicity tests of 3 species of 3 •trophic levels •1000 •100 •50 •10 •Protection level: 95% •Ecotoxicological data •Species sensitivity distribution (SSD) • •HC5 = 95% protection level •[C] •Assessment / Extrapolation factors • •PNEC 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif • Environmental quality standards / criteria Risk assessment & management • • • • ‘Hazard’ identification Data compilation Exposure assessment PEC Effect assessment PNEC Risk characterisation HQ = PEC/PNEC •< 1 > 1 •Hazard quotient •HQ Risk management 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Results of ecotoxicology • •WHAT IS IT GOOD FOR ? • •SOLVING PRACTICAL PROBLEMS 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif http://www.scotland.gov.uk/Resource/Img/237458/0069907.jpg EQS in reality – example EU Water Framework Directive 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif List of priority compounds EU WFD (selection/examples) •Most recent (2015) 44 priority compounds (table here) + additional “watch list” à see further • 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •http://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri=uriserv:OJ.L_.2015.078.01.0040.01.ENG 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Another example where ecotoxicology results are used • • •European strategy •how to deal with chemicals •REACH •à discussed elsewhere • 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Risks of chemicals: a balancing act …. between perception, uncertainties, science and pragmatism? Final considerations 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif janus • – dna Wetlands janus • • •Cellular and molecular effects •Population/community effects Effects of chemicals 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif • – • • • •Local •Global •Value •Other Technologies •Other Laws and Regulations •Other Social Needs janus •Other Sciences Risks vs. Benefits 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif j0173962 Greenpeace International faCTORY1 EU%2520Flag • Scientists Society is a balancing act … 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Closing remarks •Ecotoxicology is exciting science! •Interface: science and society •Many opportunities •Sometimes hard work –10% inspiration and 90% „perspiration“ • •Be creative: move frontiers •Keep the purpose in mind •Be critical: do not accept perceptions as facts •Speak up: you have something to say! • unclesam 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Introduction to ecotoxicology Ludek Blaha blaha@recetox.muni.cz http://www.recetox.cz