Nova Southeastern University NSUWorks HCNSO Student Theses and Dissertations HCNSO Student Work 12-7-2015 The ffecE ts of Petroleum Pollutants on Sea Urchins Reproduction and Development Kellie C. Pelikan Nova Southeastern University, [email protected] Follow this and additional works at: https://nsuworks.nova.edu/occ_stuetd Part of the Marine Biology Commons, and the Oceanography and Atmospheric Sciences and Meteorology Commons Share Feedback About This Item NSUWorks Citation Kellie C. Pelikan. 2015. The Effects of Petroleum Pollutants on Sea Urchins Reproduction and Development. Master's thesis. Nova Southeastern University. Retrieved from NSUWorks, . (401) https://nsuworks.nova.edu/occ_stuetd/401. This Thesis is brought to you by the HCNSO Student Work at NSUWorks. It has been accepted for inclusion in HCNSO Student Theses and Dissertations by an authorized administrator of NSUWorks. For more information, please contact [email protected]. NOVA SOUTHEASTERN UNIVERSITY OCEANOGRAPHIC CENTER THE EFFECTS OF PETROLEUM POLLUTANTS ON SEA URCHINS REPRODUCTION AND DEVELOPMENT By Kellie C. Pelikan Submitted to the Faculty of Nova Southeastern University Oceanographic Center In partial fulfillment of the requirements for The degree of Master of Science with a specialty in: Marine Biology & Coastal Zone Management Nova Southeastern University December 2015 ACKNOWLEDGEMENTS Firstly, I would like to express my sincere gratitude to my advisor Dr. Nicole D. Fogarty for the continuous support of my thesis research, for her patience, motivation, and immense knowledge. Her guidance helped me in all aspects my research and the writing of this thesis. I could not have imagined having a better advisor and mentor for my thesis study. Besides my advisor, I would like to thank the rest of my thesis committee: Dr. Jose Lopez and Dr. Donald McCorquodale, for their insightful comments and encouragement. My sincere thanks also go to Dr. Patricia Blackwelder, who provided me unlimited knowledge and invaluable help with the scanning electron microscopic. I thank my fellow labmates and classmates: especially Maggie Rushmore, Justin Voss, Glenn Goodwin, Josh Stocker, Hunter Noren, Leah Harper, Ashley Dungan, and Charlie Gandia, for the many long days and nights counting endless amounts of gametes, and for all the fun we have had in the last three years. Also I thank my editing friends, Brea and Veronica, for all their valuable input and numerous edit corrections at all hours of the night. I would also like to thank my family, especially my parents and sister for their endless patience while I completed this chapter of my life. Lastly, I would like to especially thank Shelly and Jeremiah, for their endless optimism and support whenever it was needed. TABLE OF CONTENTS i LIST OF TABLES iii LIST OF FIGURES iv ABSTRACT vi CHAPTER 1: Introduction 1 1.1 The ecological effects of oil spills 1 1.2 Effects on invertebrate reproduction 5 1.3 Ecological importance of echinoids 7 1.4 Purpose 9 1.5 Objectives 9 CHAPTER 2: “The influences of petroleum pollutants on the reproductive success of two sea urchin species” 10 2.1 Introduction 10 2.2 Methodology 11 2.2.1 Species description 11 2.2.2 Collection 12 2.2.3 Water accommodated fraction preparation 13 2.2.4 Fertilization 14 2.2.5 Development and Larvae viability 15 2.2.6 Scanning electron microscope (SEM) 15 2.2.7 Statistical analysis 16 2.3 Results: Species; Lytechinus variegatus 17 2.3.1 Demographic data 17 2.3.2 Fertilization 19 2.3.3 Viability 20 2.3.4 Scanning Electron microscopy 22 2.4 Results: Species; Eucidaris tribuloides 24 2.4.1 Demographic data 24 2.4.2 Fertilization 25 2.4.3 Viability 26 2.4.4 Scanning Electron microscopy 28 2.5 Species Comparison 29 2.6 Discussion 29 2.7 Conclusion 31 i CHAPTER 3: Discussion 32 3.1 Discussion 32 3.1.2 Egg Integrity 33 3.1.3 Fertilization success 33 3.1.4 Viability 35 3.2 Conclusion 36 APPENDICES: Supplementary Material 38 REFERENCES 43 ii LIST OF TABLES Table 1.1 Port Everglades Waterborne Commerce Chart, FY 2011-2002 Table 2.1 ANCOVA results of fertilization crosses for each treatment, Lytechinus variegatus iii LIST OF FIGURES Figure 1.1 Bilge water Figure 2.1 Experiment 1: Lytechinus variegatus fertilization as a function of log sperm among treatments Figure 2.2 Experiment 1: Lytechinus variegatus fertilization control and treatment comparison Figure 2.3 Lytechinus variegatus larvae survival comparison between experiments Figure 2.4 Experiment 1: Lytechinus variegatus larval survivorship Figure 2.5 Experiment 2: Lytechinus variegatus larval survivorship Figure 2.6 Comparing the developmental stages between the two experiments for Lytechinus variegatus Figure 2.7 Lytechinus variegatus development comparison between experiments Figure 2.8 Lytechinus variegatus eggs distribution in qualitative categories Figure 2.9 Lytechinus variegatus eggs categorized by damage sustained Figure 2.10 Eucidaris tribuloides control and treatment fertilization comparison Figure 2.11 Experiment 1: Eucidaris tribuloides fertilization Figure 2.12 Experiment 2: Eucidaris tribuloides fertilization Figure 2.13 Eucidaris tribuloides larvae survival comparison between experiments Figure 2.14 Comparing the developmental stages between the two experiments for Eucidaris tribuloides Figure 2.15 Eucidaris tribuloides development comparison between experiments Figure 2.16 Eucidaris tribuloides eggs distribution in qualitative categories Figure 2.17 Eucidaris tribuloides eggs categorized by damage sustained Figure 2.18 Comparison of overall survivorship between the two urchin species Figure A.1 Dispermic sea urchin egg with Aberrant development Figure A.2 Sex Ratio for Lytechinus variegatus Figure A.3 Sex Ratio for Eucidaris tribuloides Figure A.4 Average Test Size of Lytechinus variegatus iv Figure A.5 Average Test Size of Eucidaris tribuloides Figure A.6 Spawning individuals among Lytechinus variegatus Figure A.7 Spawning individuals among Eucidaris tribuloides v Abstract Disturbances, such as mass pollution events, threaten the health of vulnerable ecosystems. Recent media attention has focused on the devastating mass oil spills, but daily petroleum input from recreational and commercial ship bilge release has been overlooked. The focus of this study was the effect of petroleum products found in bilge water on fertilization success and larval viability of two sea urchin species, Lytechinus variegatus and Eucidaris tribuloides. Unlike other pollutant studies that have focused on sperm characteristics and concentrations, I chose to examine how egg integrity was compromised by petroleum products. Scanning electron microscopy revealed eggs were degraded when exposed to low levels of these pollutants. Of the three, oil was the most detrimental to Lytechinus variegatus fertilization, while gas was highly detrimental to Eucidaris tribuloides . Dosing the eggs for only two hours before introducing sperm demonstrated significant reduction in fertilization and larval survivorship. These data suggest that even relatively low and brief exposure to petroleum pollutants can have devastating effects on sea urchin reproductive success. New regulations may need to be considered when determining the safe petroleum concentration in bilge discharge. Keywords : Echinoids, Toxicity, Fertilization, Petroleum, SEM vi The influences of petroleum pollutants on the reproductive success of two sea urchins species Chapter 1 1.1 The ecological effects of oil spills The ecological significance of oil spills are complex and can have long lasting and persistent effects (Woodley et al. 1978). Lethal toxic effects for adult marine organisms when exposed to high concentrations of soluble hydrocarbons range from 1- 100 ppm, while the larval stages are significantly more sensitive with a range beginning as low as 0.01 ppm (Moore & Dwyer 1974). Oil spills often contaminate the sediments and can impact the benthic invertebrates for decades (Teal & Howarth 1984). In recent decades, millions of gallons of oil have been accidently spilled worldwide. To give a few examples, Ixtoc 1 Oil Well in the Bay of Campeche, Mexico spilled 450,000 to 1.40 million gallons of oil in 1979. The same year, the SS Atlantic Express, a Greek oil tanker, collided with another oil tanker and spilled 88.3 million gallons near Trinidad and Tobago. The Exxon Valdez oil spill occurred on March 24, 1989 when a tanker ran aground on the Bligh Reef in the Prince William Sound, Alaska, spilling approximately 11 million gallons of crude oil (Tietenberg and Lewis 2009). In 1991, during the Gulf War in Kuwait, 240-336 million gallons entered the Persian Gulf. Most recently, the Deepwater Horizon rig explosion released 210 million gallons of oil into the Gulf of Mexico on April 20, 2010. Subsequently, 1.84 million gallons of dispersants were applied to dissipate the oil (Cleveland 2010, Dept. of Interior 2012). This was the largest marine oil spill in history. Only 33 million gallons were collected in cleanup efforts (Dorsett 2010). The oil gushed from approximately 5,000 feet below the surface creating underwater plumes of oil throughout the water column and rose to create oil slicks on the surface (Soysa et al. 2012). Despite the environmental damages that oil spills may cause, the long term harm to marine organisms is unclear. After the Exxon oil spill occurred, it was assumed that the oil would dissipate over time and the affected ecosystems would recover. This unfortunately was not the case. Twelve years after the spill, Jeffrey Short, a chemist with NOAA, sampled 91 beaches in the vicinity of Prince William Sound and an estimated 1 15,000 gallons remained in the intertidal zones mostly below the rocky surface (Short et al. 2004). Twenty years later, the new estimate of remaining oil is 16,000 gallons, even more than Short’s prediction (Bernton 2009). The rate of dissipation was rapid during the first 3.5 years after the spill, but the rate of dissipation has slowed drastically since then (Short et al. 2004). The residual oil has impacted the salmon and the herring fisheries the greatest (Ott 2009).