THE EVOLUTIONARY POTENTIAL OF A PARASITE HOST: DIVERSITY, SYSTEMATICS AND PHYLOGEOGRAPHY OF THE FRESHWATER SNAILS GENUS TAREBIA H. & A. ADAMS, 1854 By MISS Nuanpan VEERAVECHSUKIJ A Thesis Submitted in Partial Fulfillment of the Requirements for Doctor of Philosophy BIOLOGY Department of BIOLOGY Graduate School, Silpakorn University Academic Year 2018 Copyright of Graduate School, Silpakorn University ววิ ฒั นาการความเกี่ยวเนื่องระหวา่ งโฮสตก์ บั ปรสิต: ความหลากหลาย ซิสเทมาติคส์ และ ชีวภูมิศาสตร์ ของหอยน้า จืดสกลุ Tarebia H. & A. Adams, 1854 โดย นางสาวนวลพรรณ วีระเวชสุกิจ วทิ ยานิพนธ์น้ีเป็นส่วนหน่ึงของการศึกษาตามหลกั สูตรปรัชญาดุษฎีบณั ฑิต สาขาวิชาชีววิทยา แบบ 1.1 ปรัชญาดุษฎีบัณฑิต ภาควิชาชีววิทยา บัณฑิตวิทยาลัย มหาวิทยาลัยศิลปากร ปีการศึกษา 2561 ลิขสิทธ์ิของบณั ฑิตวทิ ยาลยั มหาวิทยาลัยศิลปากร THE EVOLUTIONARY POTENTIAL OF A PARASITE HOST: DIVERSITY, SYSTEMATICS AND PHYLOGEOGRAPHY OF THE FRESHWATER SNAILS GENUS TAREBIA H. & A. ADAMS, 1854 By MISS Nuanpan VEERAVECHSUKIJ A Thesis Submitted in Partial Fulfillment of the Requirements for Doctor of Philosophy BIOLOGY Department of BIOLOGY Graduate School, Silpakorn University Academic Year 2018 Copyright of Graduate School, Silpakorn University 4 Title The evolutionary potential of a parasite host: diversity, systematics and phylogeography of the freshwater snails genus Tarebia H. & A. Adams, 1854 By Nuanpan VEERAVECHSUKIJ Field of Study BIOLOGY Advisor Duangduen Krailas Graduate School Silpakorn University in Partial Fulfillment of the Requirements for the Doctor of Philosophy Dean of graduate school (Associate Professor Jurairat Nunthanid, Ph.D.) Approved by Chair person (Assistant Professor Supanyika Sengsai , Ph.D.) Advisor (Associate Professor Duangduen Krailas , Ph.D.) Co Advisor (Professor Matthias Glaubrecht , Ph.D.) Examiner ( Wivitchuta Dechruksa , Ph.D.) External Examiner (Assistant Professor Col. Tunyarut Koonchornboon , Ph.D.) D ABST RACT 56303802 : Major BIOLOGY Keyword : Shell morphology, Genotypic variation, Phenotypic variation, Reproductive strategy, Trematoda, Cercariae, Intermediate host MISS NUANPAN VEERAVECHSUKIJ : THE EVOLUTIONARY POTENTIAL OF A PARASITE HOST: DIVERSITY, SYSTEMATICS AND PHYLOGEOGRAPHY OF THE FRESHWATER SNAILS GENUS TAREBIA H. & A. ADAMS, 1854 THESIS ADVISOR : ASSOCIATE PROFESSOR DUANGDUEN KRAILAS, Ph.D. Tarebia sp. is widespread and abundant in various lentic and lotic water bodies in Southeast Asia, with its range extending onto islands in Indo-West-Pacific. This snail is one of snail in family Thiaridae, as one of the most frequent and major primary intermediate host, an important vector for digenic trematodes causing several animal and human diseases. In Thailand Tarebia has been reported with the occurrence of one species only and have highly variations of shell morphology, which makes it difficult to identify them using only shell morphology. In this study, the snail samples were collected from 90 locations in Thailand, between 2014-2016. They were separated by distint shell, three groups called morphs A, B and C here, without implying morphotypes in the sense of species under a respective species concept, but for convenience only and to facilitate further research into the potential correlation of phenotypical and genetic proprinquity. Thai specimens were compared with specimens from the Centre of Natural History (CeNak), Zoological Museum, Universität Hamburg, Germany (including 12 locations from Timor-Leste). The objective of this study was integrate evidence of phylogeographical analyses based on phenotypic variation (shell morphology, using biometry and geometrical morphometrics) and genotypic variation from the gene fragments cytochrome C oxidase I and ribosomal 16S (using several phylogenetic analyses, including haplotype networks and a dated molecular tree). The results showed that Tarebia snails were found variation of both phenotype and genotype. The biometric and geometric morphometric analyses and reproductive strategy of difference morphs and genetic clades were found similarity and widely overlap. The phylogenetic trees were found two genetically distinct clades (clade A and B) and hint at possible species differentiation within what has been traditionally considered as T. granifera. All specimens from Timor-Leste were included in clade A together with specimens mostly from the southern to southern-central parts of Thailand. The clade B specimens were more frequent in the northern part of Thailand. These two lineages started to split about 5 Mya. The collected snails were investigated for cercarial infections by using shedding and crushing methods. The infection rate was 5.80% (493/8,493). The cercariae were categorized into eleven species and seven types, viz. (i) virgulate xiphidiocercariae (Loxogenoides bicolor and Loxogenes liberum), (ii) armatae xiphidiocercariae cercariae (Maritreminoides caridinae and Maritreminoides obstipus); (iii) parapleurophocercous cercariae (Haplorchis pumilio, Haplorchis taichui and Stictodora tridactyla), (iv) pleurophocercous cercariae (Centrocestus formosanus), (v) megarulous cercariae (Philophthalmus gralli), (vi) Echinostome cercariae and (vii) Gymnocephalous cercariae. In addition, a phylogenetic marker (internal transcribed spacers II, ITS2) was employed in generic and infrageneric level classifications of these trematodes. Thus, this analysis combines the parasites data on morphology and geographical occurrence with molecular phylogeny, aiming to provide the groundwork for future studies looking into more details of the parasite-snail evolutionary relationships. E ACKNOWLEDGEMENTS ACKNOWLEDGEMENTS I would like to thanks Assoc. Prof. Dr. Duangduen Krailas, who is my major advisor for her kindness, comments, suggestions, and support me throughout the course of study. I also would like to Thanks my co-advisor, Prof. Dr. Matthias Glaubrecht for him gave comment and suggestion for checked, experiment and corrected the mistake of thesis and paper until successful. I would like to special thanks for Assist. Prof. Col. Dr. Tunyarut Koonchornboon, who is the external examiner of my thesis defense for her kindness, suggestions, valuable comments and thesis correction. Also, I would like to thanks Assist. Prof. Dr. Supanyika Sengsai and Dr. Wivitchuta Dechruksa for their all of valuable suggestions, comments and reading the manuscript of this thesis. I wish to thanks Dr. Marco Thomas Neiber, Mrs. Katharina Gebauer and Miss Sarah Maria Gaugel for teaching a technique about molecular laboratory to me through support for the tips and hints of phylogenetic tree. Also, I would like to thanks for Mrs. Nora Lentge-Maaß and Mr. Benedikt Wiggering for teaching and assistance the data about geometric morphometrics. I would like to thanks SM1. Somphob khamsangeinm and Sgt. Parinya Chaiwchan for collection specimens in Thailand. In particular, thanks Mrs. Suluck Namchote for teaching a technique about identification of trematodes. I wish to thanks for all staff at Parasitology and Medical Malacology Research Unit, Silpakorn University, Thailand and Centre for Natural History (CeNak), Zoological Museum, University Hamburg, Germany for their helpful in providing facilities and materials for my thesis experiments. Thanks the collection of the Zoological Museum, University Hamburg (ZMH) and Department of Biology, Faculty of Science, Silpakorn University (SUT). I also thank the Department of Biology, Faculty of Science, Silpakorn University. I am grateful for financial support from the Thailand Research Fund through the Royal Golden Jubilee Ph. D. Program (Grant No. PHD/0093/2556) and the Deutsche Akademische Austauschdienst (DAAD) to Nuanpan Veeravechsukij, Duangduen F Krailas and Matthias Glaubrecht. The study was also supported through a collaboration grant from the Deutsche Forschungsgemeinschaft (DFG) to MG, which is thankfully acknowledged here. Finally, I would like to express my deepest gratitude to my parents and friend for their love, help, encouragement and support me complete this study. This thesis was supported by the Research and Development Institute, Silpakorn University and funded by Academic Services Deposit (Intensive English Training Program) Graduate School, Silpakorn University, Fiscal year 2016. Nuanpan VEERAVECHSUKIJ G TABLE OF CONTENTS Page ABSTRACT .................................................................................................................. D ACKNOWLEDGEMENTS .......................................................................................... E TABLE OF CONTENTS .............................................................................................. G List of tables ................................................................................................................... I List of figures ................................................................................................................. J CHAPTER I ................................................................................................................... 1 Introduction ................................................................................................................ 1 CHAPTER II ................................................................................................................ 12 Objective .................................................................................................................. 12 CHAPTER III .............................................................................................................. 13 Materials