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NOAA FISHERIES Initial assessment of the bioavailability of PAHs in Powder River Basin coal to fish early life history stages John Incardona Ecotoxicology Program NOAA Northwest Fisheries Science Center, Seattle John Incardona Ecotoxicology Program NOAA Northwest Fisheries Science Center, Seattle Slide 2 Fish embryos are sensitive indicators of PAH bioavailability crude oil bunker fuel creosote coal tar in sediments diesel exhaust particles coal tar pavement sealants stormwater runoff coal and coal dust? Slide 3 No fish species escapes PAH toxicity pink salmon (Oncorhynchus gorbuscha) Pacific herring (Clupea pallasi) Atlantic herring (Clupea harrengus) mummichog (Fundulus heteroclitus) Gulf killifish (Fundulus grandis) crimson-spotted rainbowfish (Melanotaenia fluviatilis) medaka (Oryzias latipes) zebrafish (Danio rerio) olive flounder Japanese sea perch bluefin tuna yellowfin tuna yellowtail amberjack red drum Atlantic haddock pink salmon (Oncorhynchus gorbuscha) Pacific herring (Clupea pallasi) Atlantic herring (Clupea harrengus) mummichog (Fundulus heteroclitus) Gulf killifish (Fundulus grandis) crimson-spotted rainbowfish (Melanotaenia fluviatilis) medaka (Oryzias latipes) zebrafish (Danio rerio) olive flounder Japanese sea perch bluefin tuna yellowfin tuna yellowtail amberjack red drum Atlantic haddock Slide 4 Methodologies are tailored to specific PAH sources Slide 5 Slide 6 The aryl hydrocarbon receptor pathway and cytochrome P4501A induction after Whitlock 1999, Annu Rev Pharmacol Toxicol 39:103 Slide 7 Windsor Aguirre Slide 8 Stage-specific toxicity of individual PAHs via distinct mechanisms 24 hpf 48 hpf 72 hpf 120 hpf 1.25 hpf pattern formation organogenesis hatching feeding AHR-independent cardiac dysfunction (via ion channel blockade?), CYP1A protective 36 hpf 48 hpf AHR-dependent cardiac dysfunction via inhibition of cardiomyocyte proliferation, CYP1A protective 96-120 hpf Systemic toxicity dependent on hepatic AHR and CYP1A activity Slide 9 3-ring PAH exposure phenocopies a cardiac function mutant crude oil56 M phenanthrene silent heart (troponin T) GC-MS analysis of PAHs in embryos exposed to crude oil naphthalene fluorene dibenzothiophene phenanthrene Slide 10 Slide 11 chrysenebenz(a)anthracenepyrene CYP1A IF cyp1a QPCR (TaqMan) PAHs vary in potency of CYP1A induction Slide 12 Phenanthrene is toxic with minimal CYP1A induction CYP1A MHC phenanthrenesolvent control OA CrDI 48 hpf Slide 13 Tricyclic PAHs cause bradycardia and AV conduction block heart rate (bpm) ppm phenanthrene Slide 14 Quantification of edema by pericardial area crude oil dose response (PAH) Slide 15 Scoring edema presence by pulse Slide 16 What we have tested so far Powder River Basin coal (Boardman Plant, Portland General Electric) Pulverized coal dust slurries (static exposures) Coal dust slurry filtrates (static) Coal chunk column (continuous flow) Slide 17 coal dust slurry 1% agarose embryo agarose pore size = 0.1-0.3 m embedded embryos exposed to diffusible dissolved PAH only Simultaneous exposure to either coal dust or dissolved components only Slide 18 coal chunk column