SPIRORCHIID TREMATODES OF SEA TURTLES IN FLORIDA: ASSOCIATED DISEASE, DIVERSITY, AND LIFE CYCLE STUDIES By BRIAN ADAMS STACY A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY UNIVERSITY OF FLORIDA 2008 1 © 2008 Brian Stacy 2 To my family 3 ACKNOWLEDGMENTS This project would not have been possible without the substantial contributions and support of many agencies and individuals. I am grateful for the encouragement and assistance provided by my committee members: Elliott Jacobson, Ellis Greiner, Alan Bolten, John Dame, Larry Herbst, Rick Alleman, and Paul Klein. The sea turtle community, both government agencies and private, were essential contributors to the various aspects of this work. I greatly appreciate the contributions of my colleagues in the Florida Fish and Wildlife Conservation Commission (FWC), both present and past, including Allen Foley, Karrie Minch, Rhonda Bailey, Susan Shaf, Kim Sonderman, Nashika Brewer, and Ed DeMaye. Furthermore, I thank the many participants in the Sea Turtle Stranding and Salvage Network. I also am grateful to members of the turtle rehabilitation community, including the faculty and staff of The Turtle Hospital, Mote Marine Laboratory and Aquarium, Marinelife Center at Juno Beach, the Marine Science Center, Clearwater Marine Aquarium, and the Georgia Sea Turtle Center. Specifically, I would like to thank Richie Moretti, Michelle Bauer, Corrine Rose, Sandy Fournies, Nancy Mettee, Charles Manire, Terry Norton, Ryan Butts, Janine Cianciolo, Douglas Mader, and their invaluable support staff. Essential to the life cycle aspects of this project were the critical input and collaboration of the members and volunteers of the In-water Research Group, including Michael Bressette, Blair Witherington, Shigatomo Hirama, Dean Bagley, Steve Traxler, Richard Herren, and Carrie Crady. I also thank the staff and board of the Cayman Turtle Farm, Limited for their hospitality and collaboration in life cycle studies. Joe Parsons, Catherine Bell, Brian Andrysak, and Nancy Andrysak were essential for field efforts on Grand Cayman. In addition, I am grateful to Mario Santoro and Terry Work for providing input and specimens from their considerable projects in the Costa Rica and Hawaii, respectively. 4 I am deeply indebted to Tom Frankovich who has been an essential collaborator and good friend throughout this experience. None of the field studies would have been possible without his expertise, enthusiasm, and field savvy. I also thank our volunteer divers Carlos, Mike, and Benny for their time and effort. Friends and colleagues at the University of Florida, College of Veterinary Medicine have provided support and productive collaboration for many aspects of this work. I thank Hendrik Nollens, Jim Wellehan, April Childress, Toni McIntosh, Michael Sapper, Charlie Courtney and Ruth Francis-Floyd. My family has been their throughout the long journey through undergraduate school, veterinary school, residency, and graduate school. Each has provided their own unique contributions, for which I am ever grateful. I thank my parents, James and Sandy, and my siblings, Jim, Jennifer, Chad, Brad, Marc, and Stacy for their friendship and unwavering encouragement. Furthermore, the achievements over the last four years would not have been possible without the commitment and dedication of my wife Nicole. Portions of this research were funded by the Florida Sea Turtle Tag Grants Program (2006 and 2007) and Awards 2003-0206-011 and 2006-0087-004 from the National Fish and Wildlife Foundation using funding from the National Oceanic and Atmospheric Administration. All sea turtle-related studies were performed under the authorization of the NMFS permit No. 086 and under University of Florida IACUC Project No. D994. Gastropod collections from protected areas were conducted under permissions and permits granted by the Marine Conservation Commission (Grand Cayman Island), U.S Fish and Wildlife Service (Key West National Wildlife Refuge), and National Park Service (Everglades National Park, permit no. EVER-2007-SCI-0055). 5 TABLE OF CONTENTS page ACKNOWLEDGMENTS ...............................................................................................................4 LIST OF TABLES...........................................................................................................................9 LIST OF FIGURES .......................................................................................................................11 ABSTRACT...................................................................................................................................17 CHAPTER 1 INTRODUCTION ..................................................................................................................19 Background.............................................................................................................................19 Goals and Objectives ..............................................................................................................20 Review of the Literature .........................................................................................................21 Sea Turtle Biology: Florida Species Records, Life History, and Diet ...........................21 Spirorchiid Trematodes ...................................................................................................22 Family diagnosis ......................................................................................................22 Taxonomy and phylogeny........................................................................................23 Marine spirorchiids ..................................................................................................24 Prevalence, microhabitat, and host pathology..........................................................27 Spirorchiids and serological studies.........................................................................36 Pathology associated with freshwater spirorchiids ..................................................37 Fibropapillomatosis and spirorchiids .......................................................................38 Life cycles ................................................................................................................39 Molecular studies of digeneans and applications to spirorchiid research ................44 2 SPIRORCHIID TREMATODES OF STRANDED SEA TURTLES IN FLORIDA AND ASSOCIATED PATHOLOGY ....................................................................................57 Introduction.............................................................................................................................57 Materials and Methods ...........................................................................................................58 Necropsies and Parasite Collection .................................................................................58 Parasitological Methods ..................................................................................................59 Laboratory Methods ........................................................................................................60 Categorization of Duration of Illness ..............................................................................60 Primary Diagnosis / Cause of Death Designations..........................................................61 Spirorchiid Impact Rating ...............................................................................................62 Body Condition Calculations for Immature C. mydas ....................................................62 Statistical Methods ..........................................................................................................63 Results.....................................................................................................................................63 Caretta caretta.................................................................................................................63 Study population demographics, duration of illness categories...............................63 Primary diagnosis or cause of death by duration of illness......................................64 6 Prevalence by genus and identifiable species (category 1 & 2 data). ......................66 Site predilection and associated pathological lesions ..............................................67 Spirorchiid impact rating..........................................................................................75 Chelonia mydas ...............................................................................................................77 Study population demographics and duration of illness categories .........................77 Primary diagnosis or cause of death by duration of illness......................................77 Prevalence by genus and identifiable species (category 1 & 2 data). ......................78 Site predilection and associated pathological lesions ..............................................78 Spirorchiid impact rating..........................................................................................82 Body condition indices in hypothermic-stunned immature C. mydas .....................83 Discussion...............................................................................................................................83 Conclusions...........................................................................................................................100