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Cassidy Thesis AN ABSTRACT OF THE THESIS OF Katelyn M. Cassidy for the degree of Master of Science in Fisheries Science presented on December 5, 2008. Title: Use of Extractable Lipofuscin as an Age Biomarker to Determine Age Structure of Ghost Shrimp ( Neotrypaea californiensis ) Populations in West Coast Estuaries Abstract approved: ___________________________________________________________ Brett R. Dumbauld Christopher J. Langdon Determining age in crustaceans is inherently imprecise because they molt periodically and do not retain hard structures throughout their lifespan. Morphological measurements, such as carapace length, are often used to estimate age because methods for direct ageing do not exist. However, variability in individual growth rate and molt frequency can result in a wide distribution of sizes in a single age class, making length a poor predictor of true age. Research examining the autofluorescent age pigment, lipofuscin, suggests that concentration of the pigment in neural tissues is directly related to actual age. Analysis of lipofuscin concentration has already proven to be a more effective method for determining true age in several species of crustaceans than traditional, length-based methods. The ghost shrimp, Neotrypaea californiensis , negatively impacts oyster aquaculture in Pacific Northwest estuaries. Current efforts to develop an integrated pest management plan for this species would benefit from better information on the age and growth of these animals. This study 3 assessed the potential of using extractable lipofuscin as a method for determining age in N. californiensis . A growth study was conducted to validate the lipofuscin aging technique and develop a practical method of age determination for this species . Lipofuscin-based aging was used to determine age structures for three populations of N. californiensis and these were compared to age structures determined using traditional length-based methods. Age structures determined with analysis of lipofuscin concentration revealed several more age classes than assessments based on carapace length measurements in all sampled populations. Comparison of mean size- at-age among populations in Oregon and Washington estuaries showed growth rate varied spatially. Site-specific environmental factors like food availability, population density and tidal elevation may affect individual and population growth patterns. In this study, analysis of extractable lipofuscin proved to be a more accurate method of age determination than body-length measurements. The data presented here show that biochemical-based aging can now be widely used to assess age in N. californiensis , facilitating more in-depth investigations of basic biological and ecological processes for this species. ©Copyright by Katelyn M. Cassidy December 5, 2008 All Rights Reserved Use of Extractable Lipofuscin as an Age Biomarker to Determine Age Structure of Ghost Shrimp ( Neotrypaea californiensis ) Populations in West Coast Estuaries by Katelyn M. Cassidy A THESIS Submitted to Oregon State University in partial fulfillment of the requirements for the degree of Master of Science Presented December 5, 2008 Commencement June 2009 Master of Science thesis of Katelyn M. Cassidy presented on December 5, 2008 . APPROVED: ____________________________________________________________________ Co-Major Professor, representing Fisheries Science ____________________________________________________________________ Co-Major Professor, representing Fisheries Science ____________________________________________________________________ Head of the Department of Fisheries and Wildlife ____________________________________________________________________ Dean of the Graduate School I understand that my thesis will become part of the permanent collection of Oregon State University libraries. My signature below authorizes release of my thesis to any reader upon request. ____________________________________________________________________ Katelyn M. Cassidy, Author ACKNOWLEDGEMENTS I would like to express my sincere appreciation to my major advisor, Dr. Brett Dumbauld. Thank you for providing me with all the resources I needed to complete this project. Your undying support, encouragement, patience and guidance has been my driving force over the last three years. Thank you for being a true inspiration and teaching me to love all things that live in the mud. Thanks to my other committee members: Chris Langdon for keeping me on track, and Tony D’Andrea for always reminding me to look at the big picture. Thanks to Lee McCoy, for not only being my technical and mechanical support, but also a friend. Thanks Lee for taking the monotony out of fieldwork and always making me laugh. I would like to recognize Cara Fritz for helping with processing samples and being a second set of hands in the hatchery; Roy Hildenbrand for driving the boat and helping in the field; and all others that I have dragged out on the tideflat to assist with sampling. Digging holes in the mud is a tough job. My sincere appreciation to Dr. Jim Dombrowki of the USDA-ARS Forage Seed and Cereal Unit for the use of your autosampler over last three years; and the employees of the USEPA Coastal Ecology Branch in Newport, OR, especially Bob Randall for always making sure that I had everything I need and Jim Power for the use of the spectrophotometer. Also, thanks to Se-Jong Ju, who answered my questions and provided me guidance when I was struggling along in the early stages of this project.; Pat Clinton of EPA and Keven Bennett of the Olympic Natural Resources Center for providing tidal elevation data for my study sites. Thanks to my mother and father, who always believed that I could do anything and for their unconditional love and support. 8 Thanks to Marisa Litz, for joining me in charging Oregon waves over the last three years and Tristan Britt for reminding me to relax and have fun. And finally, thanks to Keith Bosley for being my number one fan and teaching me that you can have everything you want in life. None of this work would have been possible without the help of all the people mentioned here. This work was supported by the USDA–ARS and the 2007 and 2008 Mamie Markham Awards. CONTRIBUTION OF AUTHORS Dr. Brett R. Dumbauld provided recruitment and long-term monitoring data for chapters 2 and 3. He also reviewed and provided comments on Chapters 1, 2 and 3. Drs. Christopher J. Langdon and Anthony D’Andrea reviewed and provided comments on chapters 1, 2, and 3. TABLE OF CONTENTS Page Chapter 1: General introduction and overview …........................................ 1 General biology and ecology of the ghost shrimp, Neotrypaea californiensis.................................................................................... 1 Pitfalls in assessing age with length-based methods ........................ 5 Overview of lipofuscin and its use as an age biomarker .................. 6 Ghost shrimp as pests for commercial oyster culture ...................... 11 Significance of research ................................................................... 14 LITERATURE CITED ..................................................................... 16 Chapter 2: Verification of lipofuscin-based age determination for the ghost shrimp, Neotrypaea californiensis .......................................... 26 ABSTRACT ..................................................................................... 26 INTRODUCTION ............................................................................ 28 MATERIALS AND METHODS...................................................... 30 RESULTS ........................................................................................ 37 DISCUSSION .................................................................................. 40 LITERATURE CITED..................................................................... 49 Chapter 3: Assessment of population age structure for the ghost shrimp, Neotrypaea californiensis, in West Coast estuaries using extractable lipofuscin as an age biomarker ...................... 68 ABSTRACT ..................................................................................... 68 INTRODUCTION ............................................................................ 70 MATERIALS AND METHODS...................................................... 72 RESULTS ....................................................................................... 79 11 TABLE OF CONTENTS (Continued) Page DISCUSSION.................................................................................. 83 LITERATURE CITED..................................................................... 93 Chapter 4: Summary and future directions.................................................... 120 LITERATURE CITED..................................................................... 123 Bibliography ................................................................................................. 124 APPENDICES............................................................................................... 137 Appendix A........................................................................................ 138 Appendix B........................................................................................ 140 Appendix C........................................................................................ 143 Appendix D....................................................................................... 145 LIST OF FIGURES Figure Page 1.1 Long-term
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