Biologiske effekter av veiavrenning hvordan knytte eksponering til miljørisiko? Knut Erik Tollefsen 1,2 1 NIVA-Norwegian Institute for Water Research, Oslo (NO) 2 UMB-University of Life Sciences (isotope lab), Ås (NO) Contact: ket@niva.no 1
Multiple pollutants & sources Metals Biocides Salt Pollutants (Halogenated) hydrocarbons Industrial chemicals Organometals 2
Key questions What is the hazard? Are we adressing the right effects? Are we targeting the right stressors? Is there a causality between exposure and effect? Do we adequately assess the risk? 4
What is the hazard? Biosphere Ecosystem Acute & Chronic tests Community Population Individual 5
Fjærmygg livssyklustest 28 dagers kronisk test av sediment-vann giftighet (ferskvann) OECD test 218 og ASTM test E1383-93 Biotilgjengelighet ivaretaes 4 replikater à 20 larver pr sedimentprøve Endepunkter: tid til første klekking, klekkesuksess (28 d), kumulativ klekkerate, kjønnsrate. Kontakt: Eivind Farmen (eff@niva.no) 6
Sub-lethal effects Exposure Interaction Response Pertubation Adverse effect Targets Receptor, Enzymes Membranes DNA Response Transcriptome Proteome Metabolome Interactome Effects Growth Development Metamorphosis Reproduction Death 7
Are we addressing the right effects? Sexual development Reproduction Death Embryo Fertilisation Fecundity Adults Larvae Morphogenesis Organ differentiation Growth Juvenile Metamorphosis Growth Organ development 8
Experimental approaches In vivo (2-100 days exposure) In vitro (1-4 days exposure) Air Control 1 ng/l 10 ng/l 100 ng/l 9
Transcriptomics Results interpretation Global gene expression Rapid response Response integration Characterise Mode of Action Identify key processes affected Bioinformatics Biostatistics Microarray analysis G C T A G C T G C T A G C T G C T A G C T G C T A G C T G C T A G C T RNA extraction Exposure study 10
Mortality SaltSmart 331 down Muscle development Organ development 636 Up Eye development Ion regulation Control 5000 Salt mg/l Salt (mg/l) 11
Are we targeting the right stressors? 12
Norby tunnel wash Wash water Ower flow 1 000 L 5 L/min Tap water (20 L/min) 5 L/min N = 30 N = 30 N = 30 N = 30 Same exposure, but the fish is held in 4 tanks to reduce density problems and minimizing handling stress! Outlet 10 L/min 600 L/time Meland, S., Farmen, E., Heier, L.S., Rosseland, B.O., Salbu, B., Song, Y., Tollefsen, K.E. (2011), Hepatic gene expression profile in brown trout (Salmo trutta) exposed to traffic related contaminants. Sci Total Environ 409, 1430-1443. 13
Causality between exposure & effect Environmental sample Biological analysis Chemical analysis Fractionation Biological analysis Responsible toxicant 14
Toxicity of Road-side snow Methodological approach Bioassay - Toxic potency Chemical characterisation Muusse, M., Langford, K., Tollefsen, K.E., Cornelissen, G., Haglund, P., Hylland, K., Thomas, K.V. (In press). Characterization of AhR agonist compounds in road-side snow. Analytical and Bioanalytical Chemistry. 15
Do we adequately assess the risk? Comp A Comp B Comp C Comp D.. Additivity (1+1=2) Antagonism (1+1<2) Synergy (1+1>>2) 16
Tire additives Napthenic acids (NA) Single NA 2-compound mixture 6-compound mixture Combined toxicity 2-compounds 6-compounds Prediction modelling Tollefsen, K.E., Petersen, K., Rowland, S. (In press). Toxicity of synthetic Naphthenic Acids and mixtures of these to fish liver cells. Environ. Sci. Technol. http://dx.doi.org/10.1021/es204124w 17
Combined toxicity (EDCs) Petersen and Tollefsen (2011). Assessing combined toxicity of estrogen receptor agonists in a primary culture of rainbow trout (Oncorhynchus mykiss) hepatocytes. Aquatic Toxicology 101, 186-195 18
Environmental risk assessment Exposure Effect Challenge: how to implement sub-lethal (long term) and combined effects in risk assessment! 19
Summary/recommendations Emissions from road-related activities are complex and often poorly characterised Potential adverse effects and especially sub-lethal effects are often unknown Chemical and ecotox assessment are required for thorough risk assessment High-throughput screening approaches may provide initial prioritation tools Sensitive species and life-stages should be identified Use of ecotoxcicological test batteries should be considered Causal links between exposure and effects should be established Risk assessment should consider combined toxicity and sub-lethal effects 20
Acknowledgements Financial contribution Statens Vegvesen NFR-project 178621 MixTox NFR-project 17451 UCM ERC Adv. Inv. Grant 228149 Ministry of Environment (Institutional funding) Infrastructure funding NFR-AViT 183929 Coworkers Merete Kleiven (UMB) Urma Mahrosh (UMB) You Song (UMB/NIVA) Karina Petersen (NIVA/UiO) Martine Muuse (NIVA/UiO) Eivind Farmen (NIVA) Lene Heier (UMB) Hans Christian Teien (UMB) Sondre Meland (Statens Vegvesen/UMB) Gerard Cornelissen (NGI/UoS) Peter Haglund (UoO) Kathy Langford (NIVA) Ketil Hylland (UiO) Bjørn Olav Rosseland (UMB) Brit Salbu (UMB) Kevin V. Thomas (NIVA) Steven J. Rowland (UoP) 21