1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif 1212570_28446780.jpg logo_mu_cerne.gif trojlogo.gif Ecotoxicology HAZARDS & RISKS Ludek Blaha + ecotox colleagues 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Take home messages from this presentation •Hazards are properties, while Risks are probabilities that hazards actually happen •Hazards are primarily estimated by ecotoxicity assays (resulting in LCx, NOEC,LOEC) –Regulatory example shown (including requests for standardization - OECD guidelines): REACH •The results are further used to derive PNEC (dividing tox results by uncertainty factors) •PNEC can be used as a basis for development of regulatory values (limits such as EQS) •Risks are most commonly assessed by dividing EXPOSURE data (e.g. measured concentrations or PEC values) with NO-EFFECT limit (e.g PNEC or EQS) –Regulatory example shown: EQS for priority pollutants in the EU Water Framework Directive • • • 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 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 https://encrypted-tbn3.gstatic.com/images?q=tbn:ANd9GcS52E4zw_1IWgj6a-zMCfagRjY0GxrrXUj7RPtc7h6W2mD _trk2 http://www.lfu.bayern.de/analytik_stoffe/biol_analytik_toxizitaetstests/pic/159498_gr.jpg 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif ASSESSMENT and MANAGEMENT of RISKS isk ASSESSMENT MANAGEMENT 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif WHAT IS HAZARD ? RISK? risk 1 HAZARDS (Inherited properties of stressors … including chemicals) RISKS (Probability of the actual occurrence of hazard under specific situation) 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif (Eco)Toxicology – science of „doses“ •‘What is there which is not a poison? • „Cause-effect paradigm“ •All things are poison and nothing without poison. •Solely the dose determines that a thing is not a poison. • 2 Paracelsus (1493 - 1541) 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Toxicology – ultimate goal ? To identify (or predict) safe vs hazardous levels 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif „magnitude“ or „impact“ of actual risk https://encrypted-tbn3.gstatic.com/images?q=tbn:ANd9GcQE8RGQKzHMgAfHMhqd5VcCAt-CsWdYxc7G4xMMeKH8vJ6 Suj7a Vs. Example - „P“ (probability) is the same for both events - impact is very different In ecotoxicology – e.g. fish species: 10% risks of malformations … is compensated by large numbers of eggs 10% risks of feminization à much large impact on population 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 RETROSPECTIVE PROSPECTIVE Bioassessment Field assessment Monitoring Bioassessment Field assessment Monitoring Lab studies BIOASSAYS Simulated small ecosystems DISASTERS PREDICTIONS for future Time: NOW ! Ecotoxicology: what approaches are available? Most common in practice Lab studies BIOASSAYS 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif http://3.bp.blogspot.com/-dokK9_P3A4I/UZ38uN8AoUI/AAAAAAAAAJc/d1N8vDjsmAw/s1600/what+is+a+chemical+ reactions.jpg http://upyourtalent.com/wp-content/uploads/2012/04/white-rat.jpg https://encrypted-tbn3.gstatic.com/images?q=tbn:ANd9GcS52E4zw_1IWgj6a-zMCfagRjY0GxrrXUj7RPtc7h6W2mD _trk2 http://www.lfu.bayern.de/analytik_stoffe/biol_analytik_toxizitaetstests/pic/159498_gr.jpg Organism Chemical Adverse Effects Death Altered Reproduction Inhibition of Growth Tumorigenicity Skin irritation … + Traditionally – Evaluation of adverse effects using the whole organism models Hazard assessment REGULATORY FOCUS (APICAL ENDPOINTS) Výsledek obrázku pro cyp19 Výsledek obrázku pro caco2 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif AcuteToxScheme_RM Exposure to toxicant (for defined time) Effective doses Effective concentrations (Eco)Toxicology methods 1 - standardized assays http://upyourtalent.com/wp-content/uploads/2012/04/white-rat.jpg 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 50 100 LD50 LC50 [concentration] in mg/L or % effluent Threshold: TOXICOLOGY No Obervable Adverse Effect Level (NOAEL) ECOTOXICOLOGY No Observed Effect Concentration (NOEC) Laboratory data and results http://upyourtalent.com/wp-content/uploads/2012/04/white-rat.jpg How are LC50 from ecotox assays used ? 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 Extrapolation for Risk Assessment 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% (Eco)Toxicological data Species sensitivity distribution (SSD) HC5 = 95% protection level [C] Assessment / Extrapolation factors PNEC 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Regulatory approach: risk assessment and management Hazard Exposure assessment Dose (or PEC) Effect assessment TDI (or PNEC) Risk characterisation RQ = Dose / TDI (or PEC / PNEC) RQ < 1 RQ > 1 Risk management 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Hazard Exposure assessment Dose (or PEC) Effect assessment TDI (or PNEC) Risk characterisation RQ = Dose / TDI (or PEC / PNEC) § EU Directive 98/83/EC (in addition to others) pesticide in drinking water DW in city of Bruno … atrazine 0.15 mg/L No pesticide in DW >0.1 mg/L RQ < 1 RQ > 1 Risk management RQ = 0.15/0.1 = 1.5 DWTP company $$ for penalty $$ for DWTP improvement $$ lobbying to affect legislation Regulatory approach: risk assessment and management 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif http://navier.engr.colostate.edu/whatische/images/ChEL02t01f01big.jpg • Industrial chemicals • • Cosmetics • PPP (pesticides) • Biocides • • Human pharmaceuticals • • Veterinary pharmaceuticals • Chemical laws („bulk“) REACH (ECHA) PPP (EFSA) MPs (EMA) §§ §§ WFD – surface w. GWD – ground w. Air quality Food and feed Soil & Sediments Wastes SOIL AIR WATER Two §§ approaches: ØProspective (chemicals…) Ø ØRetrospective (mixtures …) 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 Results of ecotoxicology WHAT IS IT GOOD FOR ? SOLVING PRACTICAL PROBLEMS 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Example 1 Application of ecotoxicity results (ECx à EQs) in regulatory context European Water FrameworkDIRECTIVE 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 PRACTICE: Chemical measurements vs limits (EQs) 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 is required European strategy how to deal with chemicals REACH 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •REACH Registration, Evaluation and Authorisation •of Chemicals –27-2-2001: White Paper on the Strategy for Future Chemicals Policy –23-10-2003: Commission’s proposal REACH –December 2008: Pre-registration mandatory (all chemicals in EU must be registered at ECHA European Chemicals Agency (http://echa.europa.eu) 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif –> 95,000,000 known chemicals (…and counting http://www.cas.org/) –100,000 substances in EINECS (i.e. commercial use) –30,000 relevant for REACH –cc 3000 HPVCs (High Production Volume Chemicals) Existing substances and REACH http://www.sigmaaldrich.com/content/dam/sigma-aldrich/safc/safc-reach-timeline.jpg 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif REACH legislation in EU Registration, Evaluation and Authorisation and Restriction of Chemicals 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif REACH: what data type must be registered? EU%2520Flag •Physico-chemical properties, e.g.: –Vapour pressure, boiling point, Kow,… • •Human toxicology, e.g.: –Acute and chronic toxicity, skin irritation, carcinogenity,… • •Environment/ Ecotoxicological information, e.g.: –Acute and/or chronic toxicity for aquatic organisms, biodegradation, … • • 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif REACH: testing EU%2520Flag •Total costs: 2,8 to 5,6 billion € (industry pays) •Testing costs (50-60% of total): 86% for Human, 14% Ecotox 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif •Depends on legislation (… of course !) •… but current EU legislations tend to be harmonized (use similar approaches) • à example of REACH • •Assays must be STANDARDIZED for REACH should follow OECD Guidelines • •Other standardization agencies (also include toxicity tests) e.g. ISO, ASTM • What assays and how exactly? Organisation for Economic Co-operation and Development EU%2520Flag 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif OECD guidelines for testing of chemicals •5 main sections –Section 1: Physical Chemical Properties –Section 2: Effects on Biotic Systems (i.e. Ecotoxicity) –Section 3: Degradation and Accumulation –Section 4: Health Effects (i.e. Toxicity) –Section 5: Other Test Guidelines Organisation for Economic Co-operation and Development Organization for Economic Cooperation Development 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif OECD guidelines (examples – selection) Organisation for Economic Co-operation and Development Test No. 201: Alga, Growth Inhibition Test 11 July 2006 Test No. 221: Lemna sp. Growth Inhabition Test 11 July 2006 Test No. 202: Daphnia sp. Acute Immobilisation Test 23 Nov 2004 Test No. 211: Daphnia magna Reproduction Test 16 Oct 2008 Test No. 203: Fish, Acute Toxicity Test 17 July 1992 Test No. 204: Fish, Prolonged Toxicity Test: 14-Day Study 04 Apr 1984 Test No. 210: Fish, Early-Life Stage Toxicity Test 17 July 1992 Test No. 212: Fish, Short-term Toxicity Test on Embryo and Sac-Fry Stages 21 Sep 1998 Test No. 215: Fish, Juvenile Growth Test 21 Jan 2000 Test No. 229: Fish Short Term Reproduction Assay 08 Sep 2009 Test No. 230: 21-day Fish Assay 08 Sep 2009 Test No. 231: Amphibian Metamorphosis Assay 08 Sep 2009 SECTION 2 - Aquatic organisms 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif OECD guidelines (examples – selection) Organisation for Economic Co-operation and Development Test No. 401: Acute Oral Toxicity Test No. 402: Acute Dermal Toxicity Test No. 403: Acute Inhalation Toxicity Test No. 404: Acute Dermal Irritation/Corrosion Test No. 405: Acute Eye Irritation/Corrosion Test No. 406: Skin Sensitisation Test No. 407: Repeated Dose 28-day Oral Toxicity Study in Rodents Test No. 408: Repeated Dose 90-Day Oral Toxicity Study in Rodents Test No. 409: Repeated Dose 90-Day Oral Toxicity Study in Non-Rodents Test No. 410: Repeated Dose Dermal Toxicity: 21/28-day Study Test No. 411: Subchronic Dermal Toxicity: 90-day Study Test No. 412: Subacute Inhalation Toxicity: 28-Day Study SECTION 4 – Human health effects 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif Try it! … download and study your guideline for free! 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 • – Local Global Value Other Technologies Other Laws and Regulations Other Social Needs janus Other Sciences SOCIETY – RISKS vs BENEFITS (!) 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif j0173962 Greenpeace International faCTORY1 EU%2520Flag Scientists Society is a balancing act … Scientists should contribute (provide valid data) for decision making 1212569_21823227.jpg logo_mu_cerne.gif trojlogo.gif WRAP UP and take home message – part 2 •What are hazards vs risks? –Risk assessment and management? –IMPACTS of risks? Risks vs benefits? •How are the risks calculated –By comparing EXPOSURES (PEC) with HAZARDS (PNEC) •How are hazards assessed? –By toxicity assays •there are 3 most widely used! •They must be done by standardized approaches (OECD guidelines) –What are results of toxicity assays (ICx, NOEC, LOEC)? –How are the results used? •Predictions of PNEC (by application of AFs) •Regulatory acceptance à EQS •Know examples of ecotox applications –PNEC (EQs) in Water framework directive –Predictive risk assessment in REACH regulation •