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Effects of Ultraviolet Radiation on Developing Variegated EFFECTS OF ULTRAVIOLET RADIATION ON DEVELOPING VARIEGATED SEA URCHINS, LYTECHINUS VARIEGATUS by Eric Cary Tauchman B.S., The University of Wisconsin—Madison, 2001 A thesis submitted to the Department of Biology College of Arts and Sciences The University of West Florida In partial fulfillment of the requirements for the degree of Master of Science 2008 The thesis of Eric Cary Tauchman is approved: ____________________________________________ _________________ Theodore C. Fox, Ph.D., Committee Member Date ____________________________________________ _________________ Wade H. Jeffrey, Ph.D., Committee Member Date ____________________________________________ _________________ Christopher M. Pomory, Ph.D., Committee Chair Date Accepted for the Department/Division: ____________________________________________ _________________ George L. Stewart, Ph.D., Chair Date Accepted for the University: ____________________________________________ _________________ Richard S. Podemski, Ph.D., Dean of Graduate Studies Date ACKNOWLEDGEMENTS I would like to thank my advisor, Dr. Pomory, for presenting me with the opportunity to work on this thesis. He offered teachings and advice on all things science and many things not. I also had the most knowledgeable, available, and reasonable committee members a budding scientist could ask for—something I truly appreciate Drs. Fox and Jeffrey. When one takes twice as long to complete this program as expected, focus wanders and new ideas pop up. The UWF biology and even chemistry faculty displayed wonderful patience and generosity of time and resources in abetting some less- than-entirely thought out ideas on where this research could go (some of them enough to have their names on this paper). I also did a bit of teaching during my tenure at UWF. Human Anatomy and Physiology and Cell Biology were my homes away from home sometimes. I could not have been influenced by better mentors in teaching and life than Mr. Davis and Dr. Pritchard. I’ve also made some great friends in my time at UWF (aforementioned definitely not excluded). I am grateful for you. I think that’s all. kidding. I will boldly put on paper that I owe my beautiful, new wife Jenny forever. She stayed on my side for four years of grad school, then still married me. She’s also great at data entry and proofreading and knows more about UVR effects on sea urchin larvae than she ever wanted. iii TABLE OF CONTENTS ACKNOWLEDGEMENTS ............................................................................................... iii LIST OF TABLES ............................................................................................................... v LIST OF FIGURES ........................................................................................................... vi ABSTRACT ..................................................................................................................... vii INTRODUCTION ............................................................................................................... 1 METHODS .......................................................................................................................... 8 A. Collection and Maintenance of Adults ..................................................... 8 B. Gamete Extraction and Fertilization ......................................................... 8 C. Ultraviolet Exposure ................................................................................. 9 D. Phytoplankton Culture for Larval Food .................................................. 10 E. Larval Maintenance ................................................................................ 10 F. Measurement of Morphological Effects ................................................. 11 G. Induction of Settlement ........................................................................... 12 H. Experiment 1 ........................................................................................... 13 I. Experiment 2 ........................................................................................... 14 J. Experiment 3 ........................................................................................... 15 RESULTS .......................................................................................................................... 18 A. Larval Condition ..................................................................................... 18 B. Experiment 1 ........................................................................................... 18 C. Experiment 2 ........................................................................................... 21 D. Experiment 3 ........................................................................................... 27 DISCUSSION .................................................................................................................... 32 A. Experimental Outcomes .......................................................................... 32 B. Effects on Larval Post-oral Arms ........................................................... 33 C. Effects on Larval Settlement .................................................................. 35 D. Implications of UVR Effects .................................................................. 36 REFERENCES .................................................................................................................. 42 iv LIST OF TABLES 1. Number of Lytechinus variegatus Larvae Transferred to Glass Bowls by Treatment to Examine Settlement Success in Experiment 2 ......................................... 12 2. Number of Lytechinus variegatus Larvae Transferred to Glass Bowls by Treatment to Examine Settlement Success in Experiment 3 ......................................... 13 3. Total Exposure in kJ m-2 Administered to Lytechinus variegatus Larvae in Experiment 1 for 0, 45, 90, or 135 Minutes. .................................................................. 14 4. Experiment 1: Statistical Outcome of ANOVA for Linear Regression of Right and Left Post-Oral Arm Lengths of Lytechinus variegatus Larvae Exposed to UV Radiation for Different Lengths of Time at the Gastrula Stage ..................................... 21 5. Experiment 2: Statistical Outcome from One-Way ANOVA of Mean Left and Right Post-Oral Arm Lengths of Lytechinus variegatus Larvae Exposed to 30 Minutes of UV Radiation at the Blastula, Gastrula, or Pluteus Stage and an Unexposed Control ........................................................................................................ 24 6. Experiment 2: Fisher-Hayter Multiple Comparison Tests (Α = 0.01) for Differences in Mean Left and Right Post-Oral Arm Length by Day of Measure in Lytechinus variegatus Larvae Exposed to 30 Minutes of UV Radiation at the Blastula, Gastrula, or Pluteus Stage and an Unexposed Control. Different Letters Indicate Difference, A < B < C ..................................................................................... 25 7. Experiment 3: Statistical Outcome from One-Way ANOVA of Mean Left and Right Post-Oral Arm Lengths of Lytechinus variegatus Larvae Following Exposure to Natural Sunlight with UV Filters of 280 nm, 320 nm, 395 nm or Completely Covered (Control) at the Gastrula Stage. No Statistically Significant Differences Are Present .............................................................................. 31 v LIST OF FIGURES 1. Experiment 1: Right post-oral arm length (µm) of Lytechinus variegatus larvae exposed to UVR for different lengths of time at the gastrula stage by days following fertilization. .......................................................................................... 19 2. Experiment 1: Left post-oral arm length (µm) of Lytechinus variegatus larvae exposed to UVR for different lengths of time at the gastrula stage by days following fertilization. .......................................................................................... 20 3. Experiment 2: Mean right post-oral arm lengths (µm ±SE) of Lytechinus variegatus larvae exposed to UVR for 30 minutes at the blastula, gastrula, or pluteus stage and an unexposed control by days following fertilization. ..................... 22 4. Experiment 2: Mean left post-oral arm lengths (µm ±SE) of Lytechinus variegatus larvae exposed to UVR for 30 minutes at the blastula, gastrula, or pluteus stage and an unexposed control by days following fertilization. ..................... 23 5. Experiment 2: Mean percent settlement (±SE) of Lytechinus variegatus larvae 28 and 48 days following 30 minutes of UVR exposure at the blastula, gastrula, or pluteus stage and an unexposed control. ....................................................................... 26 6. Experiment 3: Mean right post-oral arm lengths (µm ±SE) of Lytechinus variegatus larvae following exposure to natural sunlight with UVR filters of 280 nm, 320 nm, 395 nm or completely covered (control) at the gastrula stage. ......... 28 7. Experiment 3: Mean left post-oral arm lengths (µm ±SE) of Lytechinus variegatus larvae following exposure to natural sunlight with UVR filters of 280 nm, 320 nm, 395 nm or completely covered (control) at the gastrula stage. ......... 29 8. Experiment 3: Mean percent settlement (±SE) of Lytechinus variegatus larvae 17 and 24 days following exposure to natural sunlight with UVR filters of 280 nm, 320 nm, 395 nm or completely covered (control) at the gastrula stage. ......... 31 vi ABSTRACT EFFECTS OF ULTRAVIOLET RADIATION ON DEVELOPING VARIEGATED SEA URCHINS,
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