Ecotoxicology Part 2 - HAZARDS & RISKS Ludek Blaha + ecotox colleagues Ecotoxicology Science of doses / concentrations HAZARDS vs RISKS Assessment of chemical hazards …to… Humans Other organisms (TOXICOLOGY) (ECOtoxicology) ASSESSMENT and MANAGEMENT of RISKS isk ASSESSMENT  MANAGEMENT  WHAT IS HAZARD ? RISK? HAZARDS (Inherited properties of stressors … including chemicals) RISKS (Probability of the actual occurrence of hazard under specific situation) Atmospheric Deposition Erosion & Runoff Untreated discharges Predicted Environmental Concentration (PEC) Laboratory (and field) studies Ecotoxicity tests WWTP effective concentrations (PNEC) Exposure (resulting from load) Effects (what exposures cause effects ?) Cause – effect  Risk assessment 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 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 µg/L No pesticide in DW >0.1 µg/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 „magnitude“ or „impact“ of actual risk 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  much large impact on population (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. Paracelsus (1493 - 1541) Toxicology – ultimate goal ? To identify (or predict) safe vs hazardous levels 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 OrganismChemical 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) Exposure to toxicant (for defined time) Effective doses Effective concentrations (Eco)Toxicology methods 1 - standardized assays 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 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 How are LC50 from ecotox assays used ? 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” ? ? 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) C. riparius S. fontanilis O. kisuth O. reinhardtii P. fluviatilis P. partenog. P. notatus H. azteca N. barablutus P. promelas D. pulex C. dubia I. punctatus D. magna O. mykiss G. pulex C. magnifica C. decisum 0 20 40 60 80 100 1 10 100 1000 dissolved copper (µg/l) cumulativedistributionfunction Hazen plotting Loglogistic fitting HC5 = 95% protection level [C] Assessment / Extrapolation factors PNEC • Industrial chemicals • Cosmetics • PPP (pesticides) • Biocides • Human pharmaceuticals • Veterinary pharmaceuticals Chemical laws („bulk“) nanonanonanonano 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:  Prospective (chemicals…)  Retrospective (mixtures …) 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)  OK ? – Real life: easy prey  population decline 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 Results of ecotoxicology WHAT IS IT GOOD FOR ? SOLVING PRACTICAL PROBLEMS Example 1 Application of ecotoxicity results (ECx  EQs) in regulatory context European Water FrameworkDIRECTIVE EQS in reality – example EU Water Framework Directive List of priority compounds EU WFD (selection/examples) Most recent (2015) 44 priority compounds (table here) + additional “watch list”  see further PRACTICE: Chemical measurements vs limits (EQs) http://eur-lex.europa.eu/legal-content/EN/TXT/PDF/?uri=uriserv:OJ.L_.2015.078.01.0040.01.ENG Another example where ecotoxicology results are used European strategy how to deal with chemicals REACH 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) > 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 REACH legislation in EU Registration, Evaluation and Authorisation and Restriction of Chemicals REACH: what data type must be registered? • 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, … REACH: testing • Total costs: 2,8 to 5,6 billion € (industry pays) • Testing costs (50-60% of total): 86% for Human, 14% Ecotox Depends on legislation (… of course !) … but current EU legislations tend to be harmonized (use similar approaches)  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? 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 Organization for Economic Cooperation Development OECD guidelines (examples – selection) 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 OECD guidelines (examples – selection) 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 Try it! … download and study your guideline for free! Risks of chemicals: a balancing act …. between perception, uncertainties, science and pragmatism? Final considerations Local Global Value Timetobenifit Other Technologies Other Laws and Regulations Other Social Needs Other Sciences SOCIETY – RISKS vs BENEFITS (!) Scientists Society is a balancing act … Scientists should contribute (provide valid data) for decision making 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  EQS • Know examples of ecotox applications – PNEC (EQs) in Water framework directive – Predictive risk assessment in REACH regulation