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Sharealike 4.0 International License. How to Cite This This work is licensed under a Creative Commons Attribution-NonCommercial- ShareAlike 4.0 International License. How to cite this thesis / dissertation (APA referencing method): Surname, Initial(s). (Date). Title of doctoral thesis (Doctoral thesis). Retrieved from http://scholar.ufs.ac.za/rest of thesis URL on KovsieScholar Surname, Initial(s). (Date). Title of master’s dissertation (Master’s dissertation). Retrieved from http://scholar.ufs.ac.za/rest of thesis URL on KovsieScholar Taxonomic status of Trichodina heterodentata Duncan, 1977 (Ciliophora: Peritrichia) using standard morphology, as well as molecular techniques by Gerhard P. de Jager DISSERTATION submitted in fulfillment of the requirements for the degree MAGISTER SCIENTIAE IN ZOOLOGY Department of Zoology and Entomology Faculty of Natural and Agricultural Sciences Supervisor: Professor L. Basson Co-supervisor: Prof. J.G. van As UNIVERSITY OF THE FREE STATE Bloemfontein, South Africa December 2017 Table of Contents LIST OF FIGURES......................................................................................................... vi LIST OF TABLES ........................................................................................................... x CHAPTER 1 - INTRODUCTION ..................................................................................... 1 CHAPTER 2 - LITERATURE REVIEW ........................................................................... 6 THE OKAVANGO RIVER SYSTEM ............................................................................ 6 The Okavango Delta ................................................................................................ 9 Okavango Panhandle ............................................................................................. 11 Nxamasere Floodplains ......................................................................................... 12 General Hydrobiology of the Nxamasere Floodplain .............................................. 14 SOUTHERN AFRICAN HOSTS ................................................................................ 16 TELEOST HOSTS ................................................................................................. 16 ANURAN HOSTS ................................................................................................... 20 THE FAMILY TRICHODINIDAE RAABE, 1959 ...................................................... 24 GENERAL INTERNAL STRUCTURE AND ORGANELLES ................................... 26 TRICHODINID REPRODUCTION .......................................................................... 32 HISTORY OF TRICHODINID RESEARCH AND DESCRIPTIONS ........................... 34 GENERAL HISTORY OF THE GENUS TRICHODINA EHRENBERG, 1838 ......... 34 HISTORY OF TRICHODINID TAXONOMY AND SPECIES TAXONOMY AND DESCRIPTIONS .................................................................................................... 36 TRICHODINID-HOST INTERACTIONS .................................................................... 46 Trichodinid-amphibian/Trichodinid-anuran relationship .......................................... 46 BACKGROUND AND REVIEW OF TRICHODINA HETERODENTATA DUNCAN, 1977........................................................................................................................... 51 MOLECULAR RESEARCH ....................................................................................... 58 THE USE OF DNA IN MODERN TAXONOMY ...................................................... 58 MOLECULAR WORK DONE ON TRICHODINIDS ................................................ 61 CHAPTER 3 - MATERIAL AND METHODS ................................................................. 63 COLLECTION OF MATERIAL ................................................................................... 63 COLLECTION SITE ............................................................................................... 63 ii RESEARCH CAMP ................................................................................................ 64 COLLECTION AND TENDING OF HOST SPECIES ............................................. 64 HOST NOMENCLATURE ...................................................................................... 68 SEASONALITY ...................................................................................................... 68 MORPHOLOGY ........................................................................................................ 72 SILVER IMPREGNATION ...................................................................................... 72 HÆMATOXOLIN STAINING .................................................................................. 72 MEASUREMENTS ACCORDING TO LOM (1958) ................................................ 73 DENTICLE DESCRIPTIONS ACCORDING TO VAN AS AND BASSON (1989) ... 74 MOLECULAR ............................................................................................................ 75 SAMPLES, STRAINS AND PLASMIDS ................................................................. 75 ISOLATION OF genomic DNA (gDNA) .................................................................. 76 CLONING ............................................................................................................... 76 CHAPTER 4 - SILVER NITRATE EXPERIMENTS ....................................................... 81 INTRODUCTION ....................................................................................................... 81 LITERATURE AND METHODOLOGY....................................................................... 82 CHEMICAL OVERVIEW ........................................................................................ 82 HISTORIC USE IN TAXONOMY ............................................................................... 83 THE DRY SILVER IMPREGNATION AS PROPOSED BY KLEIN (1958) .............. 84 MATERIAL AND METHODS ..................................................................................... 86 EXPERIMENT 1: SILVER NITRATE IMPREGNATION OF OLD SLIDES ............. 88 EXPERIMENT 2: ALBUMEN AS SUBSTRATE ..................................................... 89 EXPERIMENT 3: GELATIN AS SUBSTRATE ....................................................... 90 EXPERIMENT 4: FISH MUCUS AS SUBSTRATE ................................................ 92 EXPERIMENT 5: RADIATION ............................................................................... 93 RESULTS AND DISCUSSION .................................................................................. 95 EXPERIMENT 1: SILVER NITRATE IMPREGNATION OF OLD SLIDES ............. 95 Method A: No substrate ......................................................................................... 95 Method B: Albumen substrate ................................................................................ 95 Method C: Gelatin substrate ................................................................................... 95 Method D: Fish mucus substrates .......................................................................... 95 iii IMPREGNATION OF OLD SLIDES SUMMARY .................................................... 99 EXPERIMENT 2: ALBUMEN SUBSTRATE ......................................................... 100 EXPERIMENT 3: GELATIN SUBSTRATE ........................................................... 100 EXPERIMENT 4: FISH MUCUS SUBSTRATE .................................................... 102 SUMMARY OF SUBSTRATES AND RADIATION EXPERIMENTS .................... 104 GENERAL AND CONCLUDING REMARKS ........................................................... 106 CHAPTER 5 - RESULTS – MORPHOLOGY .............................................................. 109 HÆMATOXYLIN STAINING .................................................................................... 109 NUCLEAR APPARATUS: .................................................................................... 109 ADORAL SPIRAL ................................................................................................. 109 SILVER NITRATE IMPREGNATIONS..................................................................... 109 BIOMETRIC DATA FROM ANURAN HOSTS ...................................................... 111 BIOMETRIC DATA FROM TELEOST HOSTS ..................................................... 116 COMPARISON OF T. HETERODENTATA DUNCAN, 1977 BIOMETRIC DATA FROM ANURAN AND TELEOST HOSTS ........................................................... 119 DENTICLE DESCRIPTIONS FROM ANURAN HOSTS ....................................... 121 DENTICLE DESCRIPTIONS FROM TELEOST HOSTS...................................... 141 CHAPTER 6 - Article Format .................................................................................... 148 TITLE: Two trichodinid species, Trichodina heterodentata Duncan, 1977 and Trichodina hypsilepis Wellborn, 1969 (Oligohymenophorea: Peritrichia) re-evaluated using historical morphological data and museum type material .................................... 148 CHAPTER 7 - RESULTS – MOLECULAR ................................................................. 163 Collection and Extraction of gDNA .......................................................................... 163 PCR Amplification and Gel Electrophoresis ............................................................. 167 Cloning and Sequencing of the 18S SSU Gene ...................................................... 169 PHYLOGENY ACCORDING TO 18S SSU SEQUENCES .....................................
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