University of North Dakota UND Scholarly Commons Theses and Dissertations Theses, Dissertations, and Senior Projects January 2015 Prevalence And Distribution Of Ranavirus, Chytrid Fungus, And Helminths In North Dakota Amphibians Melanie Paige Firkins Follow this and additional works at: https://commons.und.edu/theses Recommended Citation Firkins, Melanie Paige, "Prevalence And Distribution Of Ranavirus, Chytrid Fungus, And Helminths In North Dakota Amphibians" (2015). Theses and Dissertations. 1895. https://commons.und.edu/theses/1895 This Thesis is brought to you for free and open access by the Theses, Dissertations, and Senior Projects at UND Scholarly Commons. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of UND Scholarly Commons. For more information, please contact [email protected]. PREVALENCE AND DISTRIBUTION OF RANAVIRUS, CHYTRID FUNGUS, AND HELMINTHS IN NORTH DAKOTA AMPHIBIANS by Melanie Paige Firkins BaChelor of SCienCe, Iowa State University, 2012 Master of ScienCe, University of North Dakota, 2015 A Thesis Submitted to the Graduate FaCulty of the University of North Dakota in partial fulfillment of the requirements for the degree oF Master of ScienCe Grand Forks, North Dakota DeCember 2015 i This thesis, submitted by Melanie P. Firkins in partial fulfillment of the requirements for the Degree of Master of Science from the University of North Dakota, has been read by the FaCulty Advisory Committee under whom the work has been done and is hereby approved. ____________________________________ Dr. Robert A. Newman ____________________________________ Dr. Vasyl V. TkaCh ____________________________________ Dr. JeFFerson A. Vaughan This thesis is being submitted by the appointed advisory committee as having met all of the requirements of the School of Graduate Studies at the University of North Dakota and is hereby approved. __________________________________________ Wayne Swisher Dean of the SChool of Graduate Studies __________________________________________ Date ii PERMISSION Title Prevalence and distribution of Ranavirus, chytrid fungus, and helminths in North Dakota amphibians. Department Biology Degree Master of ScienCe In presenting this thesis in partial fulfillment of the requirements for a graduate degree from the University of North Dakota, I agree that the library of this University shall make it freely available for inspeCtion. I further agree that permission for the extensive copying for sCholarly purposes may be granted by the professor who supervised my thesis work or, in her absence, by the Chairperson of the department or the dean of the School of Graduate Studies. It is understood that any copying or publiCation or other use of this thesis or part thereof for finanCial gain shall not be allowed without my written permission. It is also understood that due reCognition shall be given to me and to the University of North Dakota in any sCholarly use which may be made of any material in my thesis. Melanie Firkins DeCember 2015 iii TABLE OF CONTENTS LIST OF TABLES ............................................................................................................. vi LIST OF FIGURES ........................................................................................................... ix ACKNOWLEDGMENTS ................................................................................................. xi ABSTRACT ...................................................................................................................... xii CHAPTER I. AMPHIBIAN PARASITES AND PATHOGENS ...........................................1 INTRODUCTION .................................................................................1 Ranavirus ...................................................................................3 Chytrid fungus .........................................................................12 Helminths ................................................................................21 OBJECTIVES ......................................................................................28 LITERATURE CITED ........................................................................29 II. PREVALENCE AND GEOGRAPHIC DISTRIBUTION OF RANAVIRUS IN NORTH DAKOTA AMPHIBIANS .....................................55 ABSTRACT .........................................................................................55 INTRODUCTION ...............................................................................56 OBJECTIVES AND HYPOTHESES ..................................................57 METHODS ..........................................................................................59 RESULTS ............................................................................................67 DISCUSSION ......................................................................................78 iv LITERATURE CITED ........................................................................82 III. PREVALENCE AND GEOGRAPHIC DISTRIBUTION OF CHYTRID FUNGUS IN NORTH DAKOTA AMPHIBIANS .......................89 ABSTRACT .........................................................................................89 INTRODUCTION ...............................................................................90 OBJECTIVES AND HYPOTHESES ..................................................91 METHODS ..........................................................................................92 RESULTS ............................................................................................97 DISCUSSION ......................................................................................98 LITERATURE CITED ....................................................................... 101 IV. PREVALENCE AND GEOGRAPHIC DISTRIBUTION OF HELMINTHS IN NORTH DAKOTA AMPHIBIANS .................................. 107 ABSTRACT ........................................................................................ 107 INTRODUCTION .............................................................................. 108 OBJECTIVES AND HYPOTHESES ................................................. 109 METHODS ......................................................................................... 112 RESULTS ........................................................................................... 121 DISCUSSION ..................................................................................... 138 LITERATURE CITED ....................................................................... 143 APPENDICES .................................................................................................................. 151 v LIST OF TABLES Table Page 1. The name and definition of predictor variables used in statistiCal models ....................................................................................................................67 2. Amphibians colleCted during 2013 field season from 68 diFFerent sites ........................................................................................................................67 3. Amphibians colleCted during 2014 field season from 103 diFFerent sites ........................................................................................................................68 4. Number (N) and percentage of sites where Ranavirus was found in at least one amphibian speCimen of any speCies in eaCh eCoregion ......................69 5. Number (N) and percentage of sites where Ranavirus was found in at least one amphibian speCimen of any speCies in eaCh land use Category ..................................................................................................................70 6. Ranavirus infeCtions by amphibian speCies ...........................................................70 7. Top models (AICC < 2) for Ranavirus ocCurrenCe pooled aCross all amphibians .............................................................................................................76 8. Model averaged coeFFiCients of Ranavirus ocCurrence aCross all amphibian speCimens using an D AIC < 2 Criterion. ............................................76 9. Top models (AICC < 2) for Ranavirus ocCurrenCe in northern leopard frogs ..........................................................................................................77 10. Model averaged coeFFiCients for Ranavirus ocCurrence in northern leopard frogs using an D AIC < 2 criterion.. .........................................................78 11. Primers and probe used for real time PCR.............................................................96 12. PCR and sequencing primers for 18S, ITS, 28S, Cox1, Nad1, 12S genes C = Cestode, D = digenean, and N = nematode. ................................117 vi 13. Number and percentage of sites where Common helminths were found in at least one amphibian speCimen of any speCies per eCoregion. .............................................................................................................124 14. Number and percentage of sites where Common helminths were found in at least one amphibian speCimen of any speCies per land use. ........................................................................................................................ 125 15. Prevalence of helminth parasites at the broadest taxonomiC level in amphibian host species. ....................................................................................... 130 16. The perCentage (%) of variable