Sondre Meland Philosophiae Doctor (P Department of Plant and EnvironmentalNorwegian Sciences University of Life Sciences • Universitetet for miljø- og biovitenskap ISBN 978-82-575-0935-4 ISSN 1503-1667 Ecotoxicological Effects of Highway and h D) Thesis 2010:25 Tunnel Wash Water Runoff Økotoksikologiske effekter av vegavrenning og tunnelvaskevann Sondre Meland Philosophiae Doctor (P The Norwegian Public Roads Administration Brynsengfaret 6A, 0667 OSLO Postbox 8142 Dep, NO–0033 Oslo Phone: +47 915 02030 www.vegvesen.no, e-mail:[email protected] h D) Thesis 2010:25 Norwegian University of Life Sciences NO–1432 Ås, Norway Phone +47 64 96 50 00 www.umb.no, e-mail: [email protected] Ecotoxicological effects of highway and tunnel wash water runoff Økotoksikologiske effekter av vegavrenning og tunnelvaskevann Philosophiae Doctor (PhD) Thesis Sondre Meland Dept. of Plant and Environmental Sciences Norwegian University of Life Sciences Ås 2010 Thesis number 2010:25 ISSN 1503-1667 ISBN 978-82-575-0935-4 Preface and acknowledgements This thesis is based on research conducted throughout the years 2005 to 2010 at the Norwegian University of Life Sciences, Department of Plant- and Environmental Sciences in collaboration with the Norwegian Public Roads Administration (NPRA). The research will hopefully contribute to increased knowledge about environmental impacts regarding highway and tunnel wash water runoff, and it will provide knowledge to the scientific community as well as to the NPRA and their sectorial environmental responsibility. The ideas and the planning of the experiments were conducted by me under supervision and guidance from Professor Brit Salbu, Professor Bjørn Olav Rosseland and Professor Oddvar G. Lindholm. The doctoral thesis is based on four scientific papers which are either published or accepted for publication (Papers I, II and III) or submitted (Paper IV) to international peer reviewed journals. First of all I would like to express my gratitude to my main supervisor Brit who admirably encouraged and helped me to finalise my plans regarding getting a PhD grant from my employer (NPRA). You have through this work provided me with constructive critics and valuable comments. Bjørn Olav, a great acknowledgement to you for sharing your knowledge regarding fish physiology and toxicology. I will also thank you for giving me the opportunity to be a guest lecturer in your course in ecotoxicology. Finally, your annual Christmas dinner has certainly been a highlight through all these years. Oddvar, you were not initially involved in the PhD-planning but joined the supervisor group in 2006. I acknowledge your contribution to Paper I. I would also like to acknowledge Knut Erik Tollefsen and Eivind Farmen at the Norwegian Institute for Water Research (NIVA) for introducing me to the field of genomics and for many interesting discussions during the work with Papers II and IV. I am full of gratitude to Professor Reidar Borgstrøm who kindly shared his data from years of sampling in Årungselva. That was an important contribution to Paper I. Lene, you have been a major support during these years, both as a friend and colleague. We have had a lot of interesting and valuable discussions which certainly have improved my thesis. Thanks to all other colleagues at the IPM/isotope laboratory, it has been a great time for me! I would also like to express my gratitude to my employer, NPRA, and especially to my former and current managers, and of course, all my colleagues. Finally, and most of all, I am very grateful for all the support and love from my wife, Åsne. I could not have done this without you! 3 Abstract In Norway, the traffic loadings have shown a substantial increase during the last decades. From 1948 to 2008 the transportation load has increased from 2.5 to 60.6 million passenger km. Hence, the ever increasing traffic has without doubt a significant negative effect on the environment. For example, highway runoffs typically contain a cocktail of both organic and inorganic contaminants being able to cause detrimental effects on the aquatic ecosystem. The present thesis, which is part of the Norwegian Public Roads Administrations ongoing work with the European Water Framework Directive, has addressed questions related to ecotoxicological effects of highway runoff. In addition, manmade runoff from tunnel wash maintenance, being far less described in the scientific literature compared to natural occurring runoffs, was included. Hence, exposure characteristics (e.g. source characterisation), environmental impacts (fish toxicity) and mitigation strategies (sedimentation ponds) were essential aspects in the present thesis. The results presented in this thesis showed that runoff water, caused by precipitation as well as by manmade tunnel wash maintenance, were contaminated. According to various environmental quality standards (EQS) from Europe and North America, the water concentrations of metals such as aluminum (Al), cupper (Cu), iron (Fe), lead (Pb), zinc (Zn) and polycyclic aromatic hydrocarbons (PAH) such as pyrene, fluoranthene, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(ghi)perylene and indeno(1,2,3- cd)pyrene were of most concern. To gain knowledge about the sources of the traffic related contaminants, enrichment factors (EF) for metals and ratios between various PAHs were calculated. The calculations were based on tunnel wash water runoff as this runoff has little or no impact from other anthropogenic sources (e.g. deposition of long range air pollution or other industrial emissions). The source characterization of tunnel wash water runoff revealed that metals such as Zn, Cu and antimony (Sb) originated mostly from the vehicle (tires and brakes). The high EFs of chloride (Cl) and sodium (Na) were most likely due to road salt applications, while the low EFs for Al, barium (Ba), calcium (Ca), chromium (Cr), Fe, potassium (K), magnesium (Mg) and nickel (Ni) indicated contribution from the pavement material. Finally, the PAH ratios revealed contribution from a mix of sources such as asphalt, tires and combustion. Generally, the concentrations of both organic and inorganic contaminants were higher in untreated highway and tunnel wash water compared to concentrations measured in the pond outlets, indicating removal of contaminants from the water phase within the pond by e.g. sedimentation. However, the removal of dissolved metal species (< 0.45 µm) was less obvious, and the investigation of a tunnel wash event in the Nordby tunnel showed that 24 % of the metal masses were discharged as low molecular mass species (< 10 4 kDa). High concentrations of road salt and dissolved organic matter (DOC) together with oxygen depletion are in this respect important as they may contribute to increased transportation and remobilization of contaminants in the sedimentation ponds. In a biological context, poor removal of contaminants as LMM species is worrying as they are considered more mobile and bioavailable than contaminants associated with particles. By using brown trout (Salmo trutta) as a model organism in the toxicity assessment of highway and manmade tunnel wash water runoff, it was demonstrated that several metals, such as Al, Cu, cobalt (Co), Fe, Pb and Sb were gill reactive. However, the metal accumulation and probably the accumulation of other contaminants as well (e.g. PAHs) were most likely modified by the high concentrations of Ca and DOC in the water. Nevertheless, the accumulation of metals in gills of exposed fish most likely provoked short term biological effects manifested by increased glucose levels in blood, being a good biomarker for general stress. Typically, this was followed by a small but notable decrease in blood plasma concentrations of Cl and Na indicating impairment of the ion regulatory system. In addition, several components associated with the antioxidant defense system were triggered in liver of exposed fish. For example, a slight increase in enzymatic activity of superoxide dismutase (SOD) and catalase (CAT) together with increased concentrations of metallothionein (MT) were measured in fish exposed to highway runoff from the Skullerud junction, while a modest up-regulation of the mRNA transcriptions of the oxidative stress biomarkers thioredoxin (TRX) and γ- glutamylcysteine synthetase (GCS) were observed in fish exposed to tunnel wash water runoff from the Nordby tunnel. The fish exposed to tunnel wash water demonstrated that PAHs and/or other organic contaminants were readily bioavailable, although normally strongly attached to particles, as a significant up-regulation of the mRNA transcription of the mixed function oxidase enzyme (phase I) cytochrome P450 1A (CYP1A) was observed. In addition to confirming increased expression of the antioxidant defense system and CYP1A in tunnel wash water exposed fish, the DNA microarray analysis revealed that traffic related contaminants also could suppress several immunological processes, as well as inhibiting the cholesterol biosynthesis several hours after the exposure. The microarray analysis also indicated the presence of organophosphorus compounds (OP), due to the apparent up-regulation of the paraoxonase enzyme (PON) which is the main protector against OP mediated neurotoxicity. Finally, an apparent up-regulation of arsenite methyltransferase (AMT) indicated that metalloids such as arsenic (As) and Sb were bioreactive, despite that no accumulation of these metalloids were observed in the liver. The short term fish exposure studies were conducted in undiluted