Independent and Interacting Effects of Multiple
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A Dissertation Submitted to the Temple University Graduate Board In Partial Fulfillment of the Requirements for the Degree by Examining Committee Members: © Copyright 2020 by Alexis M Weinnig All Rights Reserved ii ABSTRACT Human population growth and global industrial development are driving potentially irreversible anthropogenic impacts on the natural world, including altering global climate and ocean conditions and exposing oceanic environments to a wide range of pollutants. While there are numerous studies highlighting the variable effects of climate change and pollution on marine organisms independently, there are very few studies focusing on the potential interactive effects of these stressors. The deep-sea is under increasing threat from these anthropogenic stressors, especially cold-water coral (CWC) communities which contribute to nutrient and carbon cycling, as well as providing biogenic habitats, feeding grounds, and nurseries for many fishes and invertebrates. The primary goals of this dissertation are to assess the vulnerability of CWCs to independent and interacting anthropogenic stressors in their environment; including natural hydrocarbon seepage, hydrocarbon and dispersant concentrations released during an accidental oil spill (i.e. Deepwater Horizon), and the interacting effects of climate change-related factors and hydrocarbon/dispersant exposure. To address these goals, multiple stressor experiments were implemented to assess the effects of current and future conditions [(a) temp: 8°C and pH: 7.9; (b) temp: 8°C and pH: 7.6; (c) temp: 12°C and pH: 7.9; (d) temp: 12°C and pH: 7.6] and oil spill exposure (oil, dispersant, oil + dispersant combined) on coral health using the CWC Lophelia pertusa. Phenotypic response was assessed through observations of diagnostic characteristics that were combined into an average health rating at four points during exposure and recovery. Regardless of environmental condition, average health significantly declined during 24-hour exposure to dispersant alone and increased temperature resulted in a delay in recovery (72 hours) from dispersant exposure. The overall gene expression patterns varied by coral colony, but the dispersant exposure elicited the strongest response. Gene ontology (GO) enrichment analysis revealed that L. pertusa likely experienced varying stages of the cellular stress response (CSR) during exposure to iii oil, dispersant, and a decrease in pH. The most severe responses were associated with the dispersant exposure including GO terms related to apoptosis, the immune system, wound healing, and stress-related responses. However, the oil exposure induced an upregulation of metabolic pathways and energy transfer but a downregulation of cell growth and development, indicating that the coral nubbins could have been reallocating resources and reducing growth to maintain cellular homeostasis. The decrease in seawater pH elicited a similar response to oil through the enrichment of terms associated with a reduction in the cell cycle and development. Interestingly, the increase in temperature did not elicit a CSR that was detectable in the gene expression data. To further investigate the influence of hydrocarbon exposure on CWCs, comparisons of gene expression profiles were conducted using Callogorgia delta colonies that live in close proximity to active hydrocarbon seepage (“seep”) areas with no current active seepage (“non-seep”) at two different sites in the Gulf of Mexico. There were fewer differentially expressed genes in the “seep” versus “non- seep” comparison (n=21) than the site comparison (n=118) but both analyses revealed GO terms indicating slight alterations in natural biological housekeeping processes, as opposed to a CSR. Our results indicate that distinct stages of the CSR are induced depending on the intensity of stress. This bolsters the idea that there is a stress response shared by all corals in response to a variety of stressors. These data provide evidence that CWCs can be more negatively impacted, both on the phenotypic and molecular levels, by exposure to chemical dispersants than to hydrocarbons alone. Gaining an understanding of how these communities respond, not only to independent stressors, but the combination of these stressors, provides vital information about how CWC communities will fair in current and future conditions. iv DEDICATION To my late grandmother, Ellen Conwell Joslin, for showing me that women belong at the forefront of Science, Math, and Technology. v ACKNOWLEDGEMENTS I cannot fully express the gratitude I have for my mentor and friend, Erik Cordes. Without his unwavering support, advice, comradery, and compassion, I would not be where I am today. Erik has not only helped me grow as a scientist, he also emphasized the importance of being a mindful global citizen and a good person. In times when I let self-doubt cloud my view, Erik was the first one to lift me back up and help me recognize how far I had come. The opportunities that Erik provided have changed my career and my life, from trips around the world to trips to the bottom of the ocean. I know this is just the beginning of many scientific collaborations and a lifelong friendship. I look forward to our future adventures together. I am also indebted to my dissertation committee for the encouragement they have provided over the years. Thank you to Robert Sanders and Rob Kulathinal for your mentorship and support, and to Cheryl Morrison who warmly welcomed me into the deep-sea coral community and who’s guidance and friendship I have been fortunate enough to treasure on land and at sea. I am eternally thankful for my family. They have always supported my dreams and aspirations of becoming a marine scientist and I would not be the person I am today without the guidance, love, and honesty from both of my parents and my sister. My mother, Gail Joslin Weinnig, instilled in me from a young age to stand up for what I believe in and to forge my own path. My father, Michael Weinnig, taught me to always go above and beyond what is asked of you and to always keeps smiling. My sister, Melissa J. Kampshmidt, has always encouraged me to pursue what I love and to not take flak from anybody. I also have to thank The Moutons for their constant support and vi providing us with a welcoming home base in New Orleans when hurricanes tried to drive us out of the Gulf of Mexico - there is no place I would prefer to ride out a storm. I feel so fortunate to have been able to call the Cordes Lab home throughout my doctoral degree. I have never worked in an environment that fostered so much laughter, collaboration, and friendship. I was lucky enough to overlap with amazing lab mates including danielle DeLeo, Carlos Gomez, Steve Auscovitch, Jay Lunden, Amanda Glazier, Abby Keller, Sam Georgian, April Stabbins, Melissa Betters, Ryan Gasbarro, and Emily Cowell. I am truly fortunate to have all of you in my life as fellow scientists and friends. I am also incredibly appreciative to the undergraduate students that assisted in my research over the years. This work would not have been possible (and much less fun) without the diligent work of Adam Hallaj, Heejin Chung, Dan Deegan, Jessi Hart, and Alex Barkman. My Temple Biology friends have played a vital role in my success and happiness over the years. I am so thankful to have had Michele Repetto, Marianna Bonfim, Diana Lopez, Sarah DeVaul Princiotta, Elizabeth Rielly, Marianne Gagnon, Nicole Mazouchova, Janne Pfeiffernberger, and many others by my side throughout the many stages of this process. In addition, I have to thank the dedicated staff of the Biology department that always kept things running smoothly, including Toni Matthews, Tania Stephens, Carla Woodson, and Sandya Verma. I thank all of my friends, including Ryan Hulett, Shayle Matsuda, Sean Edgerton, Noel Graham, and Alex McKinzie for their enduring love, support, and constructive criticism throughout the years – I would not be where I am today without you. vii Thank you to the many programs and funding sources that made it possible for me to spend 199 days at sea during this journey, and even more so, thank you to the people that made those experiences so enjoyable and memorable. I am grateful to the ship crews aboard the D/V Ocean Inspector, M/V Ocean Intervention II, M/V Ocean Project, R/V Falkor, R/V Atlantis, M/V Alucia, R/V Brooks McCall, NOAAS Ron Brown, NOAAS Okeanos Explorer, E/V Nautilus and the vehicle crews of the ROV Global Explorer, ROV Comanche, ROV SuBastian, HOV Alvin, HOV Nadir, ROV Jason, ROV Deep Discoverer, and ROV Hercules for their commitment to facilitating successful research expeditions and sample collections. I have been extremely fortunate to have worked alongside amazing scientists and people while at sea including Caitlin Adams, Amanda demopolous, Randi Rotjan, Sarah Seabrook, Sofia Ledin, Shannon Hoy, Kasey Cantwell, Jenni McClain Counts, Luke McCartin, Sam Vohsen, and many others – I thank all of you for your hard work, comradery, and the many laughs along the way. I am extremely thankful for the funding that helped to support my dissertation research from the Gulf of Mexico Research Initiative’s “Ecosystem Impacts of Oil and Gas Inputs to the Gulf” (ECOGIG) program and Temple University for funding through teaching assistantships and the Doctoral Dissertation Completion Grant. viii TABLE OF CONTENTS ABSTRACT ....................................................................................................................... iii dEdICATION ....................................................................................................................v