NOAA FISHERIES

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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
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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?

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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

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Methodologies are tailored to specific PAH sources
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The aryl hydrocarbon receptor pathway and
cytochrome P4501A induction
after Whitlock 1999, Annu Rev Pharmacol Toxicol
39103
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Windsor Aguirre
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Stage-specific toxicity of individual PAHs via
distinct mechanisms
pattern formation
1.25 hpf
AHR-independent cardiac dysfunction (via ion
channel blockade?), CYP1A protective
36 hpf
24 hpf
organogenesis
AHR-dependent cardiac dysfunction via inhibition
of cardiomyocyte proliferation, CYP1A protective
48 hpf
48 hpf
72 hpf
hatching
Systemic toxicity dependent on hepatic AHR and
CYP1A activity
96-120 hpf
120 hpf
feeding
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3-ring PAH exposure phenocopies a cardiac
function mutant
crude oil
56 µM phenanthrene
silent heart (troponin T)
fluorene
naphthalene
dibenzothiophene
phenanthrene
GC-MS analysis of PAHs in embryos exposed to
crude oil
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PAHs vary in potency of CYP1A induction
chrysene
benz(a)anthracene
pyrene
CYP1A IF
cyp1a QPCR (TaqMan)
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Phenanthrene is toxic with minimal CYP1A induction
phenanthrene
solvent control
48 hpf
CrDI
OA
CYP1A MHC
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Tricyclic PAHs cause bradycardia and AV
conduction block
heart rate (bpm)
ppm phenanthrene
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Quantification of edema by pericardial area
crude oil dose response (?PAH)
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Scoring edema presence by pulse
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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)

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Simultaneous exposure to either coal dust or
dissolved components only
agarose pore size 0.1-0.3 µm embedded embryos
exposed to diffusible dissolved PAH only
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coal chunk column