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Polycyclic Aromatic Hydrocarbon Metabolites As a Biomarker Of University of South Florida Scholar Commons Graduate Theses and Dissertations Graduate School 11-5-2014 Polycyclic Aromatic Hydrocarbon Metabolites as a Biomarker of Exposure to Oil in Demersal Fishes Following the Deepwater Horizon Blowout Susan Susan Snyder University of South Florida, [email protected] Follow this and additional works at: https://scholarcommons.usf.edu/etd Part of the Marine Biology Commons, and the Oceanography Commons Scholar Commons Citation Snyder, Susan Susan, "Polycyclic Aromatic Hydrocarbon Metabolites as a Biomarker of Exposure to Oil in Demersal Fishes Following the Deepwater Horizon Blowout" (2014). Graduate Theses and Dissertations. https://scholarcommons.usf.edu/etd/5436 This Thesis is brought to you for free and open access by the Graduate School at Scholar Commons. It has been accepted for inclusion in Graduate Theses and Dissertations by an authorized administrator of Scholar Commons. For more information, please contact [email protected]. Polycyclic Aromatic Hydrocarbon Metabolites as a Biomarker of Exposure to Oil in Demersal Fishes Following the Deepwater Horizon Blowout by Susan M. Snyder A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science with a concentration in Marine Resource Assessment Department of Marine Science College of Marine Science University of South Florida Major Professor: Steven A. Murawski, Ph.D. Dana L. Wetzel, Ph.D. Gina M. Ylitalo, M.S. Date of Approval: November 5th, 2014 Keywords: PAH, Fish, Oil Spill, DWH, Tilefish, Gulf of Mexico Copyright© 2014, Susan M. Snyder Dedication This research is dedicated to the 11 men that lost their lives during the explosion of the Deepwater Horizon rig: Jason Anderson, Aaron Dale Burkeen, Donald Clark, Stephen Ray Curtis, Gordon Jones, Roy Wyatt Kemp, Karl Kleppinger, Keith Blair Manuel, Dewey A. Revette, Shane M. Roshto and Adam Weise. Acknowledgments I would like to acknowledgement my major advisor, Dr. Steve Murawski, for his ideas, time, effort and dedication to this project and development of his students into researchers. Steve’s concept of an integrated approach to studying the impacts of the Deepwater Horizon blowout on the benthos has lead the Center for Integrated Modeling and Analysis of Gulf Ecosystems (C-IMAGE) and this study to do great research in the wake of the blowout. Steve also put in long days leading us in the fishing and sampling effort, along with planning, coordinating and executing all cruises. I would like to thank the other two members of my committee, Dr. Dana Wetzel and Gina Ylitalo. Thank you Dana for the time in your laboratory, the uninterrupted use of your HPLC and your expertise as an analytical chemist. Thank you Gina for the analysis of the 2011 samples, your guidance on method development and instrumentation, for taking the trip to attend my defense and for your expertise as an analytical chemist. I would like to acknowledge my funding sources: the Gulf of Mexico Research Initiative (GoMRI), C-IMAGE and the endowed fellowships offered at the College of Marine Science. Out on the water, there were many people that helped in cruise planning and execution, fishing and sample collection. Thank you to Steve Murawski, David Hollander, Sherryl Gilbert, Patrick Schwing and the staff of the Florida Institute of Oceanography for your efforts in planning the cruises. A big thank you to the crews of both the C/V Pisces and the R/V Weatherbird II, for your dedication to keeping us safe while at sea and the time you spent helping us collect our samples. Thank you to our fishermen, Willie, Tia, Jorge and Chris. Thank you to all the students who helped in sample collection, Liz Herdter, Kristina Deak, Amy Wallace, Sarah Grasty, Marcy Cockrell, Jacquelin Hipes, Jenny Fenton, Nirlei Hirachy, Erin Pulster, Sherryl Gilbert and the rest of the Mud and Blood crew. On land, there were many people who contributed to the success of my data analysis, development of ideas and laboratory analysis. First, my biggest thank you is to Erin Pulster. Erin dedicated hours of her time, almost daily during the laboratory analysis, to teach me instrumentation and basic bench chemistry skills. She also provided insight while trying to understand patterns and present the data. Other thanks to the Environment Chemistry Laboratory at the Northwest Fisheries Science Center, NOAA, especially Bernadita Anulacion, for processing the 2011 bile samples and helping with the inter-lab comparison. Thanks to the other members of the Murawski lab for your moral and mental support. Thank you to Sherryl Gilbert, for all the hard work that you do running C-IMAGE and your moral support. Thank you to other C-IMAGE researchers, Dr. Isabel Romero, Dr. Patrick Schwing and Dr. David Hollander for your ideas, graphics and suggestions. Thank you to Dr. David Jones and Josh Kilborn for your help with my data analysis and statistics. Thanks to Dr. Ken Able for the insight into our study species, golden tilefish. Lastly, a large thank you to the people who have supported me personally, my family, boyfriend and my friends. Thank you to my parents, especially my father, for always pushing me to excel in the sciences and to never settle for less than 100%. Thank you to my sisters for being role models and a shoulder to lean on. Thank you to my boyfriend for your support and willingness to listen. Table of Contents List of Tables iii List of Figures iv Abstract ix Introduction 1 The Deepwater Horizon Blowout 1 Polycyclic Aromatic Hydrocarbons in Fishes 2 Study Species 9 Sediment as a Route of PAH Exposure to Fishes 14 Objectives and Hypotheses 18 Methods 19 Sample Collection 19 Laboratory Analysis 21 Bile Sample Analysis 21 Quality Assurance/Quality Control 23 Data Analysis and Statistics 24 Results and Discussion 27 Biliary PAH Metabolite Concentrations 27 Species-specific Differences in Biliary PAH Metabolite Concentration 28 Temporal Variation in Biliary PAH Metabolite Concentration 34 Biological Factors Relating to PAH Disposition 40 Spatial Variation in Biliary PAH Metabolite Concentration 44 Environmental Rationale for Biliary PAH Trends 51 Distance from the Deepwater Horizon 51 PAH Concentration in Sediment 51 Sediment Type 53 Wind Direction 57 Multivariate Analyses 59 Comparison to Historical Biliary PAH Data 63 CIMAGE Disease Survey Samples 63 Gulf of Mexico Biliary PAH Data 64 International Biliary PAH Data 67 Potential Sources of PAH Pollution 70 i Potential Biological Effects 73 Future Research 76 Conclusions 79 References 85 Appendices 97 A: PAH Metabolite Results and Biometric Data for Individual Fishes 97 B: Longline Station Locations 108 ii List of Tables Table 1: Polycyclic aromatic hydrocarbons (PAHs) analyzed in demersal fish bile samples, collected in the northern Gulf of Mexico, in this study (naphthalene, phenanthrene, benzo[a]pyrene), their abbreviations, classifications based on molecular weight (LMW = low molecular weight; HMW = high molecular weight) and molecular structures. 3 Table 2: Number of bile samples analyzed for biliary polycyclic aromatic hydrocarbon metabolites, from three species of demersal fishes collected from the northern Gulf of Mexico and year collected. 22 Table 3: 2011 biliary naphthalene and benz[a]pyrene metabolite results for red snapper (n = 30) caught in the northern Gulf of Mexico. 27 Table 4: 2012 biliary naphthalene and benz[a]pyrene metabolite results for red snapper (n = 15), golden tilefish (n = 24) and king snake eel (n = 23) caught in the northern Gulf of Mexico. 27 Table 5: 2013 biliary naphthalene and benz[a]pyrene metabolite results for red snapper (n = 63), golden tilefish (n = 72) and king snake eel (n = 44) caught in the northern Gulf of Mexico. 28 Table 6: Descriptions and references for the studies used in the international biliary PAH data comparison in Figures 43 and 44. 67 iii List of Figures Figure 1: Map of the location of the Deepwater Horizon (DWH) drilling rig when the blowout occurred on April 20th, 2010, in the Gulf of Mexico. 2 Figure 2: Profile and concentration of polycyclic aromatic hydrocarbons (PAHs) and alkylated homologs (NPH = naphthalenes; FLU = fluorenes; DBT = dibenzothiophenes; ACY = acenaphthylenes; ACE = acenaphthenes; ANT = anthracenes; PHN = phenanthrenes; FLA = fluoranthenes) measured in a sample of Deepwater Horizon (DWH) crude oil taken from the blown-out Macando well, analyzed by Reddy et al. 2012. 4 Figure 3: Profile and concentration of polycyclic aromatic hydrocarbons and alkylated homologs in a sample of Deepwater Horizon (DWH) crude oil taken from the blown-out Macando well, analyzed by British Petroleum. 5 Figure 4: Schematic of a golden tilefish vertical burrow, courtesy of Dr. Ken Able. 11 Figure 5: Map of the locations of longline stations conducted in the northern Gulf of Mexico in 2011 for the analysis of bile contamination in demersal fish species. 20 Figure 6: Map of the locations of stations conducted in the northern Gulf of Mexico in 2012 and 2013 for the analysis of bile contamination in demersal fish species. 20 Figure 7: Correlation between biliary naphthalene (NPH) and phenanthrene (PHN) metabolite concentrations (p = 0.001; r = 0.92) measured in bile of three species of demersal fishes collected in the northern Gulf of Mexico, 2011 - 2013. 24 Figure 8: Correlation between biliary naphthalene (NPH) and benzo[a]pyrene (BaP) metabolite concentrations (p = 0.001; r = 0.30) measured in bile of three species of demersal fishes collected in the northern Gulf of Mexico, 2011 - 2013. 25 Figure 9: Difference in biliary naphthalene (NPH) metabolite concentrations among the three study species sampled in 2012 and 2013 in the northern Gulf of Mexico, tested by a distribution-free analysis of variance. 29 iv Figure 10: Map of longline stations in the northern Gulf of Mexico where golden tilefish and king snake eel co-occur, and red snapper and king snake eel co-occur, sampled in 2012 and 2013.
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