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The Utilization of Morphological and Genetic Diagnostic Techniques For Minnesota State University, Mankato Cornerstone: A Collection of Scholarly and Creative Works for Minnesota State University, Mankato All Graduate Theses, Dissertations, and Other Graduate Theses, Dissertations, and Other Capstone Projects Capstone Projects 2016 The Utilization of Morphological and Genetic Diagnostic Techniques for the Description of Trematode Species Collected from Waterbirds from Lake Winnibigoshish, Minnesota, USA Tyler Joseph Achatz Minnesota State University Mankato Follow this and additional works at: https://cornerstone.lib.mnsu.edu/etds Part of the Biology Commons, and the Parasitology Commons Recommended Citation Achatz, T. J. (2016). The Utilization of Morphological and Genetic Diagnostic Techniques for the Description of Trematode Species Collected from Waterbirds from Lake Winnibigoshish, Minnesota, USA [Master’s thesis, Minnesota State University, Mankato]. Cornerstone: A Collection of Scholarly and Creative Works for Minnesota State University, Mankato. https://cornerstone.lib.mnsu.edu/etds/2016/ This Thesis is brought to you for free and open access by the Graduate Theses, Dissertations, and Other Capstone Projects at Cornerstone: A Collection of Scholarly and Creative Works for Minnesota State University, Mankato. It has been accepted for inclusion in All Graduate Theses, Dissertations, and Other Capstone Projects by an authorized administrator of Cornerstone: A Collection of Scholarly and Creative Works for Minnesota State University, Mankato. The utilization of morphological and genetic diagnostic techniques for the description of trematode species collected from waterbirds from Lake Winnibigoshish, Minnesota, USA By Tyler J. Achatz A Thesis Submitted in Partial Fulfillment of the Requirements for the Degree of Masters of Science In Biology Minnesota State University, Mankato Mankato, Minnesota April 2016 04/07/16 The utilization of morphological and genetic diagnostic techniques for the description of trematode species collected from waterbirds from Lake Winnibigoshish, Minnesota, USA. Tyler J. Achatz This thesis has been examined and approved by the following members of the student’s committee. ________________________________ Advisor Dr. Robert Sorensen ________________________________ Committee Member Dr. Timothy Secott ________________________________ Committee Member Dr. David Sharlin i Abstract The utilization of morphological and genetic diagnostic techniques for the description of trematode species collected from waterbirds from Lake Winnibigoshish, Minnesota, USA. Name: Tyler J. Achatz Degree: Masters of Science Instituiton: Minnesota State University, Mankato Mankato, Minnesota 2016 Historically, morphological techniques for species identification were the leading diagnostic methodology, however, the increased usage of genetic techniques has led to a decrease in reports of morphometrics. The decrease in morphological reports increases the chance of missing diagnostic morphometrics. The three studies described herein used morphological and genetic diagnostic methods to identify trematodes from five families in order to improve genetic and morphological information for trematode species identification. The first study identified ten species of trematodes from intestines of waterbirds previously collected from Lake Winnibigoshish, Minnesota. Nine of the species were sequenced for 28S ribosomal DNA (rDNA). Two species were also examined using ITS rDNA sequences. One species was sequenced for a portion of CO1 mitochondrial DNA as well. Morphology for all nine species was reported along with one additional species identified through morphology alone. ii The second study identified morphological and genetic variation of 28S rDNA of Neopsilotrema lisitsynae from North American waterfowl along with an analysis ofultrastructure using scanning electron microscopy. This was the first report of N. lisitsynae in North America, along with identification in four new hosts. Morphometrics of North American worms were found to vary highly in comparison to the original description from Ukraine-collected worms. Additionally, three features of Neopsilotrema were shown inaccurate in some cases: tegumental spines may be absent, egg number may be greater than 5, and the ovary may be located in a dextral, sinistral or medial position relative to the body. One variable nucleotide site was identified as well. The final study identified a new species Neopsilotrema itascae from lesser scaup using identical methods as the N. lisitsynae study. Psilotrema mediopora was also reclassified based upon morphology into Neopsilotrema.All three studies reported expansions of currently described morphometrics and diagnostic genetic sequences which may be used for future work involving species diagnosis. iii Acknowledgements First and foremost, I thank Dr. Robert Sorensen for his guidance, support, and patience. His door was always open whenever I needed help in any regard. He continually supported my work and my desire to help others learn. His passion for parasitology was always contagious to the undergraduates who helped work on the project. Without his guidance and assistance, this thesis would not have been possible. I extend thanks to Dr. Timothy Secott and Dr. David Sharlin for their help throughout the project. I greatly appreciate their assistance with both technical and logical problems. I also thank Dr. Bentley for his assistance with scanning electron microscopy. I also thank Dr. Vasyl Tkach of the University of North Dakota for his assistance obtaining literature. Additionally, thank you Holly Bloom and Scott Malotka for your support and help completing the project along with all of the undergraduates who helped throughout. I thank Dana Bennett for her immense amount of help with every aspect of the project. The scale of what was accomplished would not have been possible without her assistance. I also thank Minnesota State University, Mankato for the opportunity along with funding from both the Biology and Psychology departments. In addition, I thank the Minnesota Department of Natural resources for granting permits for waterbird collection. iv Lastly, I thank my family for all of their support throughout my time at Minnesota State, University. To my father Gordon, I thank you for continually supporting my desire to learn and pursue Biology along with your guidance and love. To my late mother Nancy, I thank you for more than I could ever write. v Table of Contents Abstract ...................................................................................................................... i Acknowledgements ....................................................................................................... iii List of Tables ................................................................................................................ vii List of Figures ............................................................................................................... ix Chapter 1: Genetic and morphological description of select trematodes from waterbirds harvested at Lake Winnibigoshish, Minnesota, USA. .............. 1 Introduction ..................................................................................................................... 2 Materials and Methods .................................................................................................... 6 Parasite collection and preparation .............................................................................. 6 Molecular analysis ....................................................................................................... 8 Alignment and Phylogenetic analysis ......................................................................... 9 Results ........................................................................................................................... 10 Notes on Family Echinostomatidae Looss, 1899: ..................................................... 11 Echinoparyphium recurvatum Linstow, 1873 ........................................................... 11 Echinoparyphium speotyto Buscher, 1978 ................................................................ 13 Echinoparyphium sp. ................................................................................................. 15 Leyogonimus polyoon Braun, 1902 ........................................................................... 17 Maritrema obstipum van Cleave & Mueller, 1932 ................................................... 18 Sphaeridiotrema pseudoglobulus McLaughlin, Scott, & Huffman, 1993................. 19 Undefined Psilostomidae Species A .......................................................................... 20 Undefined Psilostomidae Species B .......................................................................... 21 Undefined Psilostomidae Species C .......................................................................... 22 Zygocotyle lunata Diesing, 1836 ............................................................................... 23 Discussion ..................................................................................................................... 24 Echinoparyphium recurvatum Linstow, 1873 ........................................................... 25 Echinoparyphium speotyto Buscher, 1978 ................................................................ 26 Echinoparyphium sp. ................................................................................................. 28 Leyogonimus polyoon Braun, 1902
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