How was the biology of a marsh fish affected by the Deepwater Horizon oil spill?
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1 How was the biology of a marsh fish affected by the Deepwater Horizon oil spill? Fernando Galvez Dept. of Biological Sciences, Louisiana State University
2 Research model Gulf Killifish (Fundulus grandis) most abundant vertebrate in coastal marshes easy to raise in captivity relatively sensitive to chemical toxicity transparent embryos extensive characterization of embryology, physiology, biochemistry, ecology, and population genetics growing genomics tool kit (genome sequence, RNA-seq, bioinformatics, morpholinos) Photo: A. Whitehead
3 Overview of Presentation
4 Experimental Questions 1. What are the physiological and molecular impacts of oil exposure at different stages of fish development? - fitness level consequences 2. What is the geographical and temporal extent of oiling in the field? 3. How does evolutionary history and exposure history affect the sensitivity of fish to DWH oil?
5 Basic Facts of the Deepwater Horizon Oil Spill Approximately 800 million liters of oil was released during 87 days Oil was spread over 180,000 km 2 Approximately 6.9 million liters of chemical dispersant was added Exxon Valdez oil spill released approximately 40 million liters
6 Maps of Shoreline Oiling Data 1,770 km of shoreline were oiled 790 km of Louisiana shoreline were oiled
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8 Sample Sites for Gulf Killifish Monitoring BSL BFP BLB MB FMA GT Grande Terre (GT), Louisiana Bay St. Louis (BSL) and Belle Fontaine Point (BFP), Mississippi Bayou La Batre (BLB), Fort Morgan (FMA) and Upper Mobile Bay (MB), Alabama
9 Track marsh fish health before, during DWH oil exposure, and during a recovery period MB FMA BSL BFP BLB GT In our sampling, GT was the only location that oil made landfall (between June 9-14, 2010). Extremely high concentrations of polyaromatic hydrocarbons in the sediment.
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12 Experimental Design 1. Measured tissue-specific responses in fieldcollected fish. Physiology and protein-level expression changes Genome-wide mrna transcript changes 2. Performed early-life exposures to field-collected waters and sediments. time-to-hatch, hatching success, gene expression. physiology
13 Genome expression: Microarray
14 Genome Expression Response in Livers of Field-Collected Gulf Killifish AHR ligand regulated Divergent gene expression in livers of GT fish Coincident with location & timing of oil AHR ligand regulated genes were differentially expressed GT BSL BFP BLB MB FMA PNAS (2012) 50:
15 CYP1A The Aryl-hydrocarbon Receptor (AhR) PAH -Biomarkers of Exposure AhR AIP Hsp90 Arnt Endoplasmic Reticulum Nuclear mrna PNAS (2012) 50:
16 AhR-mediated CYP1A Protein Expression in Gulf Killifish Gills Trip 4 8/28/2011 Dark red stain = CYP1A protein in gills from fish collected in situ Blue stain = hematoxylin (nuclei) PNAS (2012) 50:
17 Genome Expression Response in Livers of Field-Collected Gulf Killifish AHR ligand regulated HYPOTHESIS: Exposure to toxic Divergent gene expression in livers of GT fish components of oil Coincident with location & timing of oil AHR ligand regulated genes were differentially expressed GT BSL BFP BLB MB FMA PNAS (2012) 50:
18 Genome Expression Response in Livers of Field-Collected Gulf Killifish Reproduction GT BSL BFP BLB MB FMA Genes turned OFF coincident with timing/location of oiling: Zona pellucida Choriogenin Vitellogenin PNAS (2012) 50:
19 Genome Expression Response in Livers of Field-Collected Gulf Killifish Proteolysis Ligand (PAH)-activated AHR Ubiquitin-ligase activity Proteolytic degradation of sex steroid receptors Suppression of sex hormone induced gene expression GT BSL BFP BLB MB FMA PNAS (2012) 50:
20 Genome Expression Response in Livers of Field-Collected Gulf Killifish HYPOTHESIS: Proteolysis Compromised Ligand (PAH)-activated AHR reproduction Ubiquitin-ligase activity Proteolytic degradation of sex steroid receptors (altered estrogen signaling) GT BSL BFP BLB MB FMA Suppression of sex hormone induced gene expression PNAS (2012) 50:
21 Genome Expression Response in Livers of Field Collected Gulf Killifish Cardiovascular Cardiovascular GT BSL BFP BLB MB FMA PNAS (2012) 50:
22 Genome Expression Response in Livers of Field Collected Gulf Killifish Cardiovascular Cardiovascular HYPOTHESIS: Altered cardiac physiology GT BSL BFP BLB MB FMA PNAS (2012) 50:
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25 Conclusions Large signal of hydrocarbon exposure exists in Louisiana marsh fish collected in situ. CYP1a expression is highly elevated in the gills and livers of fish collected from oil-contaminated sites. Transcriptomics data suggest potential for reduced reproductive capacity and alterations in cardiovascular physiology
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27 Sampling Sites in Southern Louisiana Bay Jimmy Bay Batiste Barataria Bay Grand Terre Island, LA Gulf of Mexico
28 Sampling Sites in Southern Louisiana Bay Jimmy Bay Batiste Grand Terre Island, LA Barataria Bay Total [PAH] µg/g Gulf of of Mexico NBS WB GT GT
29 Killifish embryo exposures to field-collected sediments EST (2013) 47:
30 Killifish embryo exposures to field-collected sediments Clean 12 ppt water Sediment EST (2013) 47:
31 Effects of PAHs on Heart Function 120 Heart Rate (BPM) P 0.05 NBS2011 WB2011 GT GT2011 GT GT 2010 Sampling Locations tpahs (ng/g) 350x x x x x NBS WB GT GT NBS 2011WB 2011 GT 2011 GT 2010 Sampling Locations EST (2013) 47:
32 Hatching and tpahs Hatch Percentage Hatch NBS 2011 WB 2011 GT 2011 GT Days Post Fertilization P tpahs (ng/g) 350x x x x x NBS WB GT GT NBS 2011WB 2011 GT 2011 GT 2010 Sampling Locations EST (2013) 47:
33 Influence of Oiled Sediments on Fish Hatch Percent of Total NBS WB GT GT Unhatched Hatched Dead EST (2013) 47:
34 PAH-induced Teratogenesis Elongated Heart and Improper Folding Pericardial Edema Craniofacial Deformities Hemorrhage
35 Conclusions Exposure to field-collected sediments increases time-to-hatch and reduces the percent hatch of killifish embryos. DWH oil leads to teratogenic effects and affects cardiovascular fitness
36 DWH Oil Spill: NEXT?... Oceans and Human Health Centers: Mechanisms of reproductive, developmental, and early life stage impacts of marine Fernando Galvez Christopher Green Andrew Whitehead
37 Health risk Early-life development Juvenile/adult performance Reproduction
38 Health risk Early-life development + Juvenile/adult performance Reproduction Low level exposures during critical developmental stages impacts in later life
39 Health risk Early-life development + Juvenile/adult performance Reproduction Low level exposures during critical developmental stages impacts in later life Adult exposures impacts on reproduction
40 Health risk Early-life development + Juvenile/adult performance Reproduction Low level exposures during critical developmental stages impacts in later life Adult exposures impacts on reproduction Adult & early-life exposures trans-generational impacts
41 Health risk Early-life development Juvenile/adult performance Reproduction Low level exposures during critical developmental stages impacts in later life Adult exposures impacts on reproduction Adult & early-life exposures trans-generational impacts Mechanisms morphology, physiology, genome expression, genome imprinting
42 Health risk Early-life development Juvenile/adult performance Reproduction 1 TX 2 LA Genetic diversity 3 FL 4 LA recent exposure history 5 TX long exposure history Low level exposures during critical developmental stages impacts in later life Adult exposures impacts on reproduction Adult & early-life exposures trans-generational impacts Mechanisms morphology, physiology, genome expression, genome imprinting Genetic variation & exposure history protect or sensitize individuals
43 Gulf killifish with different exposure histories were collected along the ngom Gang s Bayou, TX 2 LSU Aquaculture, LA 3 St. Teresa, FL Reference populations 4 Grande Terre, LA (2015) prior DWH oil exposure 5 Vince Bayou, TX long exposure history
44 Gulf killifish with different exposure histories were collected along the ngom Brood fish collection Vince Bayou, TX Gang s Bayou, TX Grand Terre, LA (2015) LSU, LA Parental exposures PC PE PC PE PC PE PC PE Fertilization Each population of adult fish were exposed to control water (i.e., parental controls; PC) or to 25% highenergy oiled WAF for at least 40 days (i.e., parental exposed; PE)
45 Fertilization (%) Gulf killifish with different exposure histories were collected along the ngom Brood fish collection Vince Bayou, TX Gang s Bayou, TX Grand Terre, LA (2015) LSU, LA Parental exposures PC PE PC PE PC PE PC PE Fertilization PC PE Vince Bayou Gang's Bayou Grande Terre LSU-LA
46 Gulf killifish with different exposure histories were collected along the ngom Brood fish collection Vince Bayou, TX Gang s Bayou, TX Grand Terre, LA (2015) LSU, LA Parental exposures PC PE PC PE PC PE PC PE Fertilization Embryo DWH Oil exposures for all treatments 0% 10% 32% 56%
47 Conclusions Genetic variation & exposure history can either protect or sensitize individuals to subsequent oil exposure F1 embryos from Louisiana populations are more sensitive to DWH oil F0 adults from an adapted population are recalcitrant to DWH oil (i.e., retains high reproductive capacity F1 embryos from an adapted population is more resistant to cardiovascular teratogenesis
48 Whitehead lab Jennifer Roach Jan Park Green lab Chelsea Bonner Zachary McDaniel Galvez lab Charles Brown Tiffany Simms Chelsea Adams Ben Dubansky Acknowledgements
49 QUESTIONS?
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