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Non-Lethal Determination of Heavy Metals in Spiny Dogfish (Squalus Suckleyi) Spines Using LA-ICP-MS

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Non-Lethal Determination of Heavy Metals in Spiny Dogfish (Squalus Suckleyi) Spines Using LA-ICP-MS Western Washington University Western CEDAR WWU Graduate School Collection WWU Graduate and Undergraduate Scholarship 2012 Non-lethal determination of heavy metals in spiny dogfish (Squalus suckleyi) spines using LA-ICP-MS Clayton L. (Clayton Louis) Bailes Western Washington University Follow this and additional works at: https://cedar.wwu.edu/wwuet Part of the Marine Biology Commons Recommended Citation Bailes, Clayton L. (Clayton Louis), "Non-lethal determination of heavy metals in spiny dogfish (Squalus suckleyi) spines using LA-ICP-MS" (2012). WWU Graduate School Collection. 238. https://cedar.wwu.edu/wwuet/238 This Masters Thesis is brought to you for free and open access by the WWU Graduate and Undergraduate Scholarship at Western CEDAR. It has been accepted for inclusion in WWU Graduate School Collection by an authorized administrator of Western CEDAR. For more information, please contact [email protected]. Non-lethal determination of heavy metals in Spiny Dogfish (Squalus suckleyi) spines using LA-ICP-MS By Clayton Louis Bailes Accepted in Partial Completion Of the Requirements for the Degree Master of Science Kathleen L. Kitto, Dean of the Graduate School ADVISORY COMMITTEE Chair, Dr. Leo Bodensteiner Dr. Ruth Sofield Dr. Polly Berseth MASTER’S THESIS In presenting this thesis in partial fulfillment of the requirements for a master’s degree at Western Washington University, I grant to Western Washington University the non‐exclusive royalty‐free right to archive, reproduce, distribute, and display the thesis in any and all forms, including electronic format, via any digital library mechanisms maintained by WWU. I represent and warrant this is my original work, and does not infringe or violate any rights of others. I warrant that I have obtained written permissions from the owner of any third party copyrighted material included in these files. I acknowledge that I retain ownership rights to the copyright of this work, including but not limited to the right to use all or part of this work in future works, such as articles or books. Library users are granted permission for individual, research and non‐commercial reproduction of this work for educational purposes only. Any further digital posting of this document requires specific permission from the author. Any copying or publication of this thesis for commercial purposes, or for financial gain, is not allowed without my written permission. Clayton Bailes 20 September 2012 Non-lethal determination of heavy metals in Spiny Dogfish (Squalus suckleyi) spines using LA-ICP-MS A Thesis Presented to The Faculty of Western Washington University In Partial Fulfillment Of the Requirements for the Degree Master of Science by Clayton Louis Bailes August 2012 ABSTRACT Biological structures that develop incremental growth patterns over time present a unique opportunity to study chronological aspects of the organism’s chemical environment. Spiny Dogfish (Squalus suckleyi), an abundant shark species, develop two dorsal spines that exhibit this type of growth pattern. The growth patterns on these spines have been used extensively as indicators of age. However, the chronological patterns of trace metal deposits in these spines have yet to be assessed. The main goals of this study were to develop the methods for analyzing this chronology and to explore techniques to analyze these data. Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) is a recently developed analytical technique for studying spatially-distributed elemental compositions in solids. LA-ICP-MS was used to quantify the concentrations of zinc and strontium across the life histories of 18 Spiny Dogfish. Metal accumulation and size differed between sharks caught at two sampling locations. This method was able to chronologically relate metal deposition to age in individuals of this species. IV ACKNOWLEDGEMENTS I thank my advisor, Dr. Leo Bodensteiner, and my committee members, Dr. Ruth Sofield and Dr. Polly Berseth, for their guidance and support. Graham Bailes and Karl Mueller assisted in fishing and with sample collection. George Mustoe showed me how to cut thin sections of my spine samples. Diane Peterson, Dorene Gould and Fraser Wilkinson were always there to help me keep things going smoothly. Robin Matthews took time, on numerous occasions, to offer her thoughts on my statistical design. Sandy Rosenfield at the Washington Department of Fish and Wildlife taught me their aging methodology. Emily Linroth edited, proofread, and generally improved this thesis. My parents offered support and encouragement throughout the entire process. Finally, I thank everyone else who contributed their thoughts and time to this project. V TABLE OF CONTENTS ABSTRACT…………………………………………………………………………….IV AKNOWLEDGEMENTS………………………………………………………………V LIST OF TABLES……………………………………………………………………VIII LIST OF FIGURES…………………………………………………………………….IX LIST OF EQUATIONS……………………………………………………………….XII LIST OF ABBREVIATIONS………………………………………………………..XIII INTRODUCTION……………………………………………………………………….1 NON-LETHAL SAMPLING IN FISH…………………………………………....1 SPINY DOGFISH…………………………………………………………………1 LA-ICP-MS……………………………………………………………………….3 METAL ACCUMULATION IN FISH…………………………………………...3 AGE DETERMINATION………………………………………………………...4 RESEARCH OBJECTIVES………………………………………………………6 METHODS……………………………………………………………………………….7 SAMPLE COLLECTION…………………………………………………………7 SAMPLE PROCESSING…………………………………………………………9 AGE DETERMINATION………………………………………………….……12 LA-ICP-MS METHODOLOGY………..……………………………………….13 DATA ANALYSIS METHODOLOGY…………………………………………17 METHODS DEVELOPMENT………………………………………………………...21 LASER ABLATION SETTINGS………………………………………………..21 TRANSECT ENDPOINT DETERMINATION…………………………………21 VI DIRECTION OF SCAN…………………………………………………………24 TIME RESOLVED VERSUS SPECTRUM MODE…………………………….24 RESULTS AND DISCUSSION………………………………………………………..26 EVALUATION OF DETECTION AND QUANTITATION LIMITS…….……26 TRENDS IN ZINC AND STRONTIUM TRANSECT DATA………………….29 VARIABILITY BY SEX AND CAPTURE LOCATION………………………50 RELATION AMONG METALS, AGE AND SIZE…………………………….53 DISTRIBUTION OF METAL CONCENTRATIONS, SIZE AND AGE………55 PREDICTION OF SEX AND SAMPLE LOCATION………………………….59 CONCLUSIONS………………………………………………………………………..68 REFERENCES………………………………………………………………………….69 APPENDIX A…………………………………………………………………………...75 VII LIST OF TABLES Table 1. Detection and quantitation limits in mg/kg of chromium, copper, and zinc for individual spines based on blank and NIST 614 measures………………………………27 Table 2. Detection and quantitation limits in mg/kg of strontium, cadmium, and lead for individual spines based on blank and NIST 614 measures………………………………28 Table 3. Pairwise comparisons of mean total metal accumulation, size, and age according to sample location and sex……………………………………………………………….52 Table 4. Relations among metal concentrations, sizes, and ages………………………...53 Table 5. Kmeans cluster means by group for each of five runs.…………………………60 Table 6. Kmeans clustering of groups by sex……………………………………………61 Table 7. Kmeans clustering of groups by location………………………………………62 Table 8. Results of Pearson’s Chi-Squared Test for kmeans clustering based on metal content, length, and age………………………………………………………………….63 VIII LIST OF FIGURES Figure 1. Representation of inner dentine cones of a four-year-old fish………………….5 Figure 2. External annuli seen on the mantle of the spine………………………………...5 Figure 3. Spine cross-section mounted in epoxy resin to a frosted glass slide…………..11 Figure 4. Inner dentine annuli from a cross-sectioned spine of shark B viewed at 100 X using transmitted light.…………………………………………………………………...12 Figure 5. Laser ablation transects lines across the inner dentine of dogfish spines viewed under transmitted light………………………………………………………………...…17 Figure 6. Platinum wire wrapped around spine cross-section…………………………...23 Figure 7. LA-ICP-MS transects for zinc from shark A………………………………….31 Figure 8. LA-ICP-MS transects for zinc from shark B………………………………..…31 Figure 9. LA-ICP-MS transects for zinc from shark C………………………………….32 Figure 10. LA-ICP-MS transects for zinc from shark D………………………………...32 Figure 11. LA-ICP-MS transects for zinc from shark E………………………………....33 Figure 12. LA-ICP-MS transects for zinc from shark G………………………………...33 Figure 13. LA-ICP-MS transects for zinc from shark H………………………………...34 Figure 14. LA-ICP-MS transects for zinc from shark I………………………………….34 Figure 15. LA-ICP-MS transects for zinc from shark K………………………………...35 Figure 16. LA-ICP-MS transects for zinc from shark L…………………………………35 Figure 17. LA-ICP-MS transects for zinc from shark M………………………………...36 Figure 18. LA-ICP-MS transects for zinc from shark N………………………………...36 Figure 19. LA-ICP-MS transects for zinc from shark O………………………………...37 Figure 20. LA-ICP-MS transects for zinc from shark P…………………………………37 Figure 21. LA-ICP-MS transects for zinc from shark Q………………………………...38 IX Figure 22. LA-ICP-MS transects for zinc from shark R…………………………………38 Figure 23. LA-ICP-MS transects for zinc from shark S…………………………………39 Figure 24. LA-ICP-MS transects for zinc from all sharks and estimated corresponding dates……………………………………………………………………………………...40 Figure 25. LA-ICP-MS transects for strontium from shark A…………………………...41 Figure 26. LA-ICP-MS transects for strontium from shark B…………………………...41 Figure 27. LA-ICP-MS transects for strontium from shark C…………………….……..42 Figure 28. LA-ICP-MS transects for strontium from shark D……………………...……42 Figure 29. LA-ICP-MS transects for strontium from shark E…………………..……….43 Figure 30. LA-ICP-MS transects for strontium from shark F………………………..….43 Figure 31. LA-ICP-MS transects for strontium
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