Download (1MB)

UNIVERSITI PUTRA MALAYSIA

HEALTH EVALUATION, PARASITE DIVERSITY AND REPRODUCTIVE PROFILING IN THE CRITICALLY ENDANGERED PAINTED TERRAPIN (Batagur borneoensis, Schlegel and Muller, 1844)

VISHWANEE KOLANDAIVELOO

FPV 2019 7

HEALTH EVALUATION, PARASITE DIVERSITY AND REPRODUCTIVE PROFILING IN THE CRITICALLY ENDANGERED PAINTED TERRAPIN (Batagur borneoensis, Schlegel and Muller, 1844) UPM

VISHWANEE KOLANDAIVELOO

COPYRIGHT Thesis Submitted to School of Graduate Studies, Universiti Putra Malaysia in Fulfillment of the Requirements for the Degree of Master of © Veterinary Science January 2018 All material contained within the thesis, including without limitation text, logos, icons, photographs and all other artwork, is copyright material of Universiti Putra Malaysia unless otherwise stated. Use may be made of any material contained within the thesis for non- commercial purposes from the copyright holder. Commercial use of material may only be made with the express, prior, written permission of Universiti Putra Malaysia.

UPM

HEALTH EVALUATION, PARASITE DIVERSITY AND REPRODUCTIVE PROFILING IN THE CRITICALLY ENDANGERED PAINTED TERRAPIN (Batagur borneoensis, Schlegel and Muller, 1844)

VISHWANEE KOLANDAIVELOO January 2018 UPM Chairman : Reuben Sharma (DVM, MVSc, PhD, MRSB, CBiol) Faculty : Veterinary Medicine

Painted terrapin (Batagur borneoensis) are critically endangered, hard-shelled freshwater chelonians that inhabit rivers and estuaries of Malaysia, Brunei Darussalam, Thailand and Indonesia. The natural range of this terrapin species has reduced dramatically over the years due to pressures from environmental disturbances, habitat loss, and poaching. One of the ways to increase this species population is through captive breeding. However, there have been no successful ex situ captive breeding programmes of this species in South East Asia and this could be due to several factors like physical health, hormones, environment and nutrition. The causative factors could not be determined without conducting comparative health evaluation and reproductive profiling between captive and wild painted terrapins. Therefore the present study was conducted to evaluate the health and reproductive profile of wild B. borneoensis from Sungai Linggi, Melaka, and a captive population from a zoological facility in Melaka, Peninsular Malaysia. Blood was collected from 142 wild (50 adult male, 66 adult female, 7 sub-adult male, 11 sub-adult female, and 8 juveniles) and 12 adult (3 male and 9 female) captive painted terrapins for haematology and clinical biochemistry analyses, and monthly reproductive hormone (progesterone, oestrogen and testosterone) profiling through enzyme immunoassays (ELISA). All wild and captive painted terrapins were also subjected to morphometry, physical examination and health screening which included parasite detection and radiography. Changes in breeding colouration was monitored and scored for the males. Normal ranges for haematology and plasma biochemistry was established for the first time for wild B. borneoensis. Results indicated thatCOPYRIGHT the captive terrapins are in poorer health condition based on physical abnormalities, haematology, clinical biochemistry, and parasite burden compared to their wild conspecifics. This could be due to several factors such as stress of captivity, improper enclosure and poor nutrition. The most common lesions for the wild terrapins occurred on the carapace region (65.5%), while that of the captive individuals were found on the © limbs (100.0%). The parasites that were encountered in this species include Falcaustra (9.4%), Orientodiscus (1.9%), Ozobranchus (30.7%), and haemogregarines (45.3%), which constitutes the first report for B. borneoensis. Helminth ova burden was more

prevalent in the captive (66.7%) compared to the wild (50%) terrapins. Six wild females were found to be gravid, while only one male and one female captive terrapin showed indications of a normal breeding cycle. In the wild population, hormone profiles indicate that mating and spermiation occurs in October-November before ovulation in December- January. The breeding coloration of the wild male terrapins corresponded well with the temporal cycle of reproductive hormones. In the captive population, mating and ovulation occurs in October-November before spermiation in December-January, which explains why the eggs produced by the captive females are not fertilised. The wild population provided a baseline on the natural reproductive cycle of this species and through this study, there was asynchrony of timing between spermiation in the captive males and ovulation in the females. The reason for this hormonal imbalance could be related to suboptimal health and husbandry in captivity. It is envisaged that the data obtained will assist in the protection and conservation of this terrapin species in the wild and also assist in the management of captive individuals meant for conservation breeding in zoological facilities. UPM

Abstrak thesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai memenuhi keperluan untuk ijazah Master Sains Veterinar

PENILAIAN KESIHATAN, DIVERSITI PARASIT DAN PROFIL REPRODUKTIF DALAM TUNTUNG LAUT (Batagur borneoensis, Schlegel dan Muller, 1844) YANG HAMPIR PUPUS

Oleh

VISHWANEE KOLANDAIVELOO

Januari 2018 UPM

Pengerusi : Reuben Sharma (DVM, MVSc, PhD, MRSB, CBiol) Fakulti : Perubatan Veterinar

Tuntung laut (Batagur borneoensis) merupakan spesies chelonia air tawar yang bercangkerang keras dan hampir pupus, mendiami sungai dan muara Malaysia, Brunei Darussalam, Thailand dan Indonesia. Sejak beberapa tahun kebelakangan ini, populasi spesies ini semakin berkurangan secara mendadak disebabkan oleh gangguan persekitaran, kehilangan habitat dan pemburuan haram. Salah satu kaedah untuk meningkatkan populasi spesies ini ialah melalui program pembiakan dalam kurungan. Walaubagaimanapun, program pembiakan dalam kurungan tidak menunjukkan sebarang kejayaan di Asia Tenggara dan ini mungkin kerana beberapa faktor seperti kesihatan fizikal, hormon, persekitaran dan nutrisi. Akan tetapi, faktor penyebab utama tidak dapat dikenalpasti tanpa membuat perbandingan tahap kesihatan dan profil reproduksi antara tuntung liar dan tuntung dalam zoo. Oleh itu, kajian ini dilaksanakan bagi menilai tahap kesihatan dan profil reproduktif B. borneoensis liar dari Sungai Linggi, Melaka dan populasi dari zoo di Melaka, Semenanjung Malaysia. Sampel darah yang diperolehi daripada 142 tuntung liar (50 jantan dewasa, 66 betina dewasa, 7 sub-dewasa jantan, 11 sub-dewasa betina, and 8 juvenil) dan 12 dewasa di zoo (3 jantan dan 9 betina) digunakan untuk analisis hematologi dan biokimia klinikal, profil hormon reproduktif bulanan (progesterone, oestrogen and testosterone) melalui enzyme immunoassays (ELISA). Semua tuntung liar dan di zoo telah melalui morfometri, pemeriksaan fizikal dan kesihatan yang meliputi pengesanan parasit dan radiografi. Perubahan dalam corak pembiakan jantan dipantau dan diberi skor. Julat normal untuk hematologi dan biokimia telah dibuat kali pertama untuk tuntung liar B. borneoensis. Hasil kajian menunjukkan tahapCOPYRIGHT kesihatan tuntung di zoo kurang memuaskan berdasarkan kepada perubahan fizikal, hematologi, biokimia klinikal dan beban parasit berbanding dengan tuntung liar. Ini mungkin disebabkan oleh beberapa faktor seperti tekanan dalam kurungan, tempat tinggal yang tidak sesuai dan kekurangan nutrisi. Lesi yang paling ketara pada tuntung © liar adalah di karapas (65.5%), manakala tuntung di zoo pada kaki (100.0%). Parasit yang dijumpai pada spesis ini termasuk Falcaustra (9.4%), Orientodiscus (1.9%), Ozobranchus (30.7%), dan haemogregarines (45.3%), dan ini merupakan penemuan

iii

pertama untuk B. borneoensis. Beban telur cacing lebih ketara pada tuntung di zoo (66.7%) berbanding dengan tuntung liar (29%). Enam ekor tuntung betina didapati mempunyai telur manakala hanya satu ekor tuntung jantan dan satu ekor tuntung betina di zoo menunjukkan kitaran pembiakan normal. Dalam populasi liar, profil hormon menunjukkan proses pengawanan dan kematangan sperma berlaku pada Oktober- November before ovulasi yang berlaku pada Disember-Januari. Corak pembiakan tuntung jantan liar selaras dengan kitaran hormone reproduktif. Dalam populasi di zoo, pengawanan dan ovulasi berlaku pada Oktober-November sebelum sperma menjadi matang pada Disember-Januari, menunjukkan telur yang dihasilkan oleh tuntung betina di zoo tidak disenyawakan. Populasi tuntung liar telah menunjukkan kitaran reproduktif yang semulajadi dan melalui kajian ini didapati ketidakselarasan antara tempoh kematangan sperma dan ovulasi pada populasi di zoo. Punca ketidakseimbangan hormon muungkin disebabkan oleh kesihatan yang sub-optima dan keadaan di dalam kurungan yang kurang memuaskan. Data yang diperolehi melalui kajian ini akan membantu dalam perlindungan dan pemuliharaan tuntung liar dan membantu dalam pengurusan populasi dalam kurungan bagi tujuan pembiakan di zoo. UPM

ACKNOWLEDGEMENTS

I would like to express my deepest gratitude to my supervisor, Dr.Reuben Sharma for all his advices, guidance and moral support. Thank you for always challenging and motivating me to do better. I also would like to thank my other supervisory committee members, Prof. Madya Dr. Gurmeet Kaur and Prof. Madya Dr. Malaika Watanabe for all their help and moral support.

I would like to thank the Parasitology staff members of Faculty Veterinary Medicine, UPM, Pn. Maizatul Akmal and Mr.Rashid for their help in identification of the parasites and for their care and love. Not to forget my wonderful labmates, Ruviniyia, Syahir, Zarith, Dilaila, Dila, Dr.Gimba, Dr.Shola, Pradeep and Donea for helping me during my sampling, laboratory works and caring for the terrapins. UPM

Special thanks to WWF, Malaysia and their staff, Richard for collaborating in this project and helping me with the trapping of the terrapins and sampling. Richard, your dedication to wildlife conservation always inspires me to do more. Thank you.

I would like to thank zoo veterinarian, Dr. Boon Nie and staffs from Zoo Melaka for allowing and helping me to conduct my research there.

Nothing is impossible if you have supportive people around you and I am blessed with wonderful family (dad, mum, sis and in laws) who always pushes me forward. Thank you for always believing in me even when I don’t believe in myself. Last but not least I would like to thank my husband for being my pillar of strength. Marriage and pregnancy does not make a women weak, it makes her stronger. I am stronger now because of my husband and my daughter. I personally would to dedicate this thesis to my daughter. “Do not let anything/anyone stop you from achieving your dreams. Take one step at a time, you will eventually reach your destination.”

UPM

COPYRIGHT © This thesis was submitted to the Senate of Universiti Putra Malaysia and has been accepted as fulfillment of the requirement for the degree of Master of Veterinary Science. The members of the Supervisory Committee were as follows:

Reuben Sharma, PhD Senior Lecturer Faculty of Veterinary Medicine Universiti Putra Malaysia (Chairman)

Malaika Watanabe, PhD Associate Professor Faculty of Veterinary Medicine Universiti Putra Malaysia UPM (Member)

Gurmeet Kaur Dhaliwal, PhD Associate Professor Faculty of Veterinary Medicine Universiti Putra Malaysia (Member)

ROBIAH BINTI YUNUS, PhD Professor and Dean School of Graduate Studies Universiti Putra Malaysia

Date:

vii

Declaration by graduate student

I hereby confirm that:  this thesis is my original work;  quotations, illustrations and citations have been duly referenced;  this thesis has not been submitted previously or concurrently for any other degree at any other institutions;  intellectual property from the thesis and copyright of thesis are fully-owned by Universiti Putra Malaysia, as according to the Universiti Putra Malaysia (Research) Rules 2012;  written permission must be obtained from supervisor and the office of Deputy Vice-Chancellor (Research and Innovation) before thesis is published (in the form of written, printed or in electronic form) including books, journals, modules, proceedings, popular writings, seminar papers, manuscripts, posters, reports, lecture notes, learning modules or any other materials as stated in the UniversitiUPM Putra Malaysia (Research) Rules 2012;  there is no plagiarism or data falsification/fabrication in the thesis, and scholarly integrity is upheld as according to the Universiti Putra Malaysia (Graduate Studies) Rules 2003 (Revision 2012-2013) and the Universiti Putra Malaysia (Research) Rules 2012. The thesis has undergone plagiarism detection software.

Signature: ______Date: ______

Name and Matric No.: Vishwanee Kolandaiveloo (GS39763)

viii

Declaration by Members of Supervisory Committee

This is to confirm that:  the research conducted and the writing of this thesis was under our supervision;  supervision responsibilities as stated in the Universiti Putra Malaysia (Graduate Studies) Rules 2003 (Revision 2012-2013) are adhered to.

Signature: Name of Chairman of Supervisory Committee:

UPM

Signature: Name of Member of Supervisory Committee:

TABLE OF CONTENTS

Page ABSTRACT i ABSTRAK iii ACKNOWLEDGEMENTS v APPROVAL vi DECLARATION viii LIST OF TABLES xii LIST OF FIGURES xiii LIST OF ABBREVIATIONS xvi

CHAPTER 1 INTRODUCTION 1

2 LITERATURE REVIEW UPM Natural history and conservation of Batagur borneoensis 3 The reproductive biology of chelonians 7 Chelonian reproductive anatomy 7 Chelonian reproductive physiology 8 Secondary sexual characteristic and gender determination 11 Methods to profile reproductive cycles in chelonians 12 Radiography 12 Ultrasonography 14 Endoscopy 14 Hormone Assays 15 Health profiling in chelonians Haematology and clinical biochemistry as indicators of health 16 Parasites of chelonians: diversity and pathogenicity 21 Parasite diversity of chelonians in Malaysia Helminths 23 Arthropods and Protozoa 24

3 HEALTH EVALUATION OF WILD AND CAPTIVE Batagur borneoenis Introduction 28 Methodology Animals 29 Morphometry and physical examination 31 Sample collection 33 Haematology and plasma biochemistry 33 Parasite detection COPYRIGHTWet mount and thin blood film 33 Formal ether sedimentation 33 Identification of ectoparasites and helminths 34 Statistical analysis 34 Results © Morphometry 35 Abnormalities and deformities 35

Haematology and clinical biochemistry Effects of environment Wild adult versus captive adult 40 Wild juvenile versus captive juvenile 40 Wild adult female versus captive adult female 40 Effects of gender Wild adult male versus wild adult female 48 Wild sub-adult female versus wild sub-adult male 48 Effects of age Wild adult female versus wild sub-adult female 48 Wild adult male versus wild sub-adult male 48

Species reference values and comparative clinical pathological 49 condition between the wild and captive terrapin populations Parasitology Adult worms 59 Helminth ova UPM60 Haemoparasites 68 Ectoparasites 69 Discussion 71 Conclusion 76

4 TEMPORAL REPRODUCTIVE PROFILING OF WILD AND CAPTIVE Batagur borneoensis Introduction 77 Methodology Animals, blood collection and radiography 78 Breeding colouration score 78 Enzyme-linked immunoassay (ELISA) 81 Statistical analysis 81 Results 82 Breeding colouration score and gravid status 82 Temporal reproductive hormone profiling Wild adult male Batagur borneoensis 87 Captive adult male Batagur borneoensis 89 Wild adult female Batagur borneoensis 94 Captive adult female Batagur borneoensis 96 Discussion 101 Conclusion 106

5 CONCLUSION AND RECOMMENDATIONs 107

REFERENCES 109 COPYRIGHTBIODATA OF STUDENT 120 LIST OF PUBLICATION 121

LIST OF TABLES

Table Page

2.1 Diversity of parasites previously reported from chelonians in 25 Peninsular Malaysia.

3.1 Morphometry (mean ± SD, range in parenthesis) of Batagur 36 borneoensis from Melaka, Peninsular Malaysia according to age and gender.

3.2 Categories of external abnormalities and their prevalence in wild 39 and captive Batagur borneoensis from Melaka, Peninsular Malaysia. UPM 3.3 Haematology values (mean ± SD, range in parenthesis) for wild and 41 captive Batagur borneoensis from Melaka, Peninsular Malaysia.

3.4 Plasma biochemistry values (mean ± SD, range in parenthesis) for 44 wild and captive Batagur borneoensis from Melaka, Peninsular Malaysia.

3.5 Corrected haematologic values (mean ± SD, reference 52 interval/reference range in parenthesis) for wild and captive Batagur borneoensis from Melaka, Peninsular Malaysia, with outliers removed.

3.6 Corrected plasma biochemistry values (mean ± SD, reference 55 interval/reference range in parenthesis) for wild and captive Batagur borneoensis from Melaka, Peninsular Malaysia, with outliers removed.

3.7 Prevalence (%) of parasites in wild and captive Batagur borneoesis 63 from Melaka, Peninsular Malaysia.

3.8 Morphometry of Falcaustra sp. recovered from Batagur 65 borneoensis from Melaka, Peninsular Malaysia.

3.9 Morphometry of Orientodiscus sp. recovered from Batagur 67 borneoensis from Melaka, Peninsular Malaysia. COPYRIGHT4.1 Breeding colouration score (0 – 3) for adult male Batagur 81 borneoensis from Sungai Linggi, Melaka, Peninsular Malaysia.

xii

LIST OF FIGURES

Figure Page

2.1 Distribution range (green) of the Painted Terrapin, Batagur 4 borneoenis in Southeast Asia.

2.2 External features of Batagur borneoensis. 6

2.3 Diagnostic imaging of chelonians can assist in the reproductive 13 profiling of females.

3.1 Satellite view (Google map image 2017) of Batagur borneoensis 30 trapping stations along Sungai Linggi, Melaka, Peninsular Malaysia and its tributaries. UPM 3.2 The enclosure of captive Batagur borneoensis at a zoological 30 facility in Melaka, Peninsular Malaysia.

3.3 Morphometry measurements done on wild and captive Batagur 32 borneoensis from Melaka, Peninsular Malaysia.

3.4 Different types of lesions in Batagur borneoensis from Melaka, 38 Peninsular Malaysia.

3.5 Prevalence of physical abnormalities between captive and wild 39 Batagur borneoensis from Melaka, Peninsular Malaysia.

3.6 Comparison of haematologic pathological conditions between 58 wild and captive from Batagur borneoensis from Melaka, Peninsular Malaysia.

3.7 Comparison of clinical biochemistry pathological conditions 58 between wild and captive Batagur borneoensis from Melaka, Peninsular Malaysia.

3.8 Falcaustra sp. recovered from Batagur borneoensis from 62 Melaka, Peninsular Malaysia.

3.9 The trematode, Orientodiscus sp. recovered from Batagur 64 COPYRIGHTborneoensis from Melaka, Peninsular Malaysia 3.10 Falcaustra sp. and Trematode eggs recovered from Batagur 66 borneoensis from Melaka, Peninsular Malaysia.

3.11 Unidentified helminth ova recovered from Batagur borneoensis 67 © from Melaka, Peninsular Malaysia.

xiii

3.12 Haemogregarines recovered from wild Batagur borneoensis 69 from Melaka, Peninsular Malaysia.

3.13 The marine leech, Ozobranchus sp. recovered from wild 70 Batagur borneoensis from Melaka, Peninsular Malaysia

4.1 Breeding colouration of male Batagur borneoensis from Sungai 80 Linggi, Melaka, Peninsular Malaysia.

4.2 Number of wild Batagur borneoensis sampled bi-monthly from 83 April 2015 to March 2016 at Sungai Linggi, Melaka, Peninsular Malaysia.

4.3 Gender ratio of Batagur borneoensis sampled bi-monthly from 83 April 2015 to March 2016 at Sungai Linggi, Melaka, Peninsular UPM Malaysia.

4.4 Breeding colouration score of adult wild male Batagur 84 borneoensis by month from April 2015 to March 2016 at Sungai Linggi, Melaka, Peninsular Malaysia.

4.5 Graphic representation of the Cumulative Breeding Colouration 84 Index (CBCI) (score x percentage) of wild adult male Batagur borneoensis over a one-year period at Sungai Linggi, Melaka, Peninsular Malaysia.

4.6 Breeding colouration of one captive adult male at a zoological 85 facility in Melaka, Peninsular Malaysia followed over a period of one year.

4.7 Gravid status of adult female Batagur borneoensis by month 86 from April 2015 to March 2016 at Sungai Linggi, Melaka, Peninsular Malaysia.

4.8 Radiograph of two wild adult female Batagur borneoensis from 86 Sungai Linggi, Melaka, Peninsular Malaysia with presence of eggs in the coelomic cavity

4.9 Temporal hormone concentration (Mean ± SE) of wild adult 88 male Batagur borneoensis from Sungai Linggi, Melaka, Peninsular Malaysia.

COPYRIGHT4.10 Temporal hormone concentration (Mean ± SE) of captive adult 88 male Batagur borneoensis at a zoological facility in Melaka, Peninsular Malaysia. © 4.11 Comparison of testosterone concentration (Mean ± SE) between 90 adult male wild and captive population of Batagur borneoensis.

xiv

4.12 Individual testosterone concentration of captive male Batagur 90 borneoensis by month.

4.13 Comparison of progesterone concentration (Mean ± SE) 91 between wild and captive male Batagur borneoensis.

4.14 Individual progesterone concentration of captive male Batagur 91 borneoensis by month.

4.15 Comparison of oestrogen (Mean ± SE) concentration between 93 wild and captive male Batagur borneoensis.

4.16 Oestrogen concentration of three captive males by month. 93 4.17 Temporal hormone concentration (Mean ± SE) of wild adult UPM95 female Batagur borneoensis from Sungai Linggi, Melaka, Peninsular Malaysia.

4.18 Temporal hormone concentration (Mean ± SE) of captive adult 95 female Batagur borneoensis at a zoological facility in Melaka, Peninsular Malaysia.

4.19 Comparison of testosterone (Mean ± SE) concentration between 97 wild and captive female Batagur borneoensis.

4.20 Testosterone concentration of individual captive females by 97 month.

4.21 Comparison of progesterone (Mean ± SE) concentration 98 between wild and captive female Batagur borneoensis.

4.22 Progesterone concentration of individual captive females by 98 month.

4.23 Comparison of oestrogen (Mean ± SE) concentration between 100 wild and captive female Batagur borneoensis.

4.24 Oestrogen concentration of individual captive female by month. 100

LIST OF ABBREVIATIONS

CCL Curved carapace length

CCW Curved carapace width

SCL Straight carapace length

SCW Straight carapace width

CPL Curved plastron length

CPW Curved plastron width SPL Straight plastron length UPM SPW Straight plastron width

An-Cl Length from the anal scute to the cloaca

Cl-Tp Length from the cloaca to the tip of the tail

Pc-Ab Pectoral to abdominal scute length

WAS Width of the anal scutes

Ht Height

Wt Weight

PCV Packed cell volume

TRBC Total red blood cell count

Hb Total haemoglobin

WBC White blood cells

MCHC Mean corpuscular haemoglobin concentration

AST Aspartate aminotransferase COPYRIGHTLDH Lactate dehydrogenase ALP Alkaline phosphatase © CK Cretinine kinase TBL Total body length

xvi

GBW Greatest body width

PHL Pharynx length

PHW Pharynx width

OCL Oesophagus corpus length

OCW Oesophagus corpus width

OIL Oesophagus isthmus length

OIW Oesophagus isthmus width

OBL Oesophagus bulb length

OBW Oesophagus bulb width UPM

SPL Spicule length

SPW Spicule width

STail Spicule from the tail tip

GL Gubernaculum length

PS tail Pseudosucker from tail tip

PP Papillae pattern

VuTail Vulva from tail tip

TAL Anus from tail tip

EGGL Egg length

EGGW Egg width

OSL Oral sucker length

OSW Oral sucker width COPYRIGHTOS Ant Oral sucker from anterior extremity Ant TEL Anterior testis length

Ant TEW Anterior testis width

xvii

Post TEW Posterior testis width

PosTe Ant Position of anterior testis from anterior extremity

PosTe Post Position of anterior testis from posterior extremity

OV Ant Position of ovary from anterior extremity

OVL Ovary length

OVW Ovary width

VS Ant Ventral sucker from anterior extremity VS L Ventral sucker length UPM VSW Ventral sucker width

Bif Ant Caecal bifurcation from anterior extremity

EsoL Oesophagus length

EGGL Eggs length

EGGW Eggs width

xviii

CHAPTER 1

GENERAL INTRODUCTION

Chelonians (Testudines) which comprise the turtles, terrapins and tortoises, are believed to be the earliest evolved reptiles with fossil records dating back as early as 215 million years ago, when the dinosaurs were still alive (Hall, 2011). These ancient reptiles occupy a diverse array of niches throughout the globe including marine, freshwater, and terrestrial habitats. Chelonians that live in seawater are known as marine turtles, those found in fresh water are termed as terrapins or freshwater turtles, and the ones living predominantly on land are known as tortoises (Hall, 2011). To date 94 genera and 335 species of chelonians have been recorded globally (Turtle Taxonomy Working Group, 2014). Of these, 57 species are considered to be critically endangered, 50 species endangered, 60 species vulnerable and 39 species near threatened due to the rapidly dwindling numbers in the wild (Turtle Taxonomy Working Group, 2014). CheloniansUPM throughout the world, and especially in Asia, are under major threat due to over exploitation and habitat degradation. They are being constantly harvested from the wild to supply the pet and ornament trade, and to be used as food and traditional medicine (Turtle Conservation Coalition, 2011). Over the years, eight species and three subspecies of turtle have gone extinction (Turtle Conservation Coalition, 2011; Turtle Taxonomy Working Group, 2014).

One of the critically endangered chelonian species is the Painted terrapin (Batagur borneoensis), which is a hard-shelled species from the subfamily Batagurinae. Its extant range includes Southern Thailand, Peninsular Malaysia, Borneo and Sumatra (International Union for Conservation of Nature - IUCN, 2000; Bonin et al., 2006). Although these terrapins are spread across Southeast Asia, their actual distribution is extremely limited to only a number of large river systems. In Peninsular Malaysia, viable populations can only be found in Sungai Setiu (Terengganu), Sungai Linggi (Melaka/Negeri Sembilan) and Sungai Paka (Terengganu) (Convention on International Trade in Endangered Species of Wild Fauna and Flora - CITES, 2006). There has been a dramatic (80%) reduction in the B. borneoensis population over the last three generations causing it to be listed as critically endangered by the IUCN Red List of Threatened Species (IUCN, 2000), and under Appendix II of the Convention on International Trade of Endangered Species (CITES, 2006). Wild populations of this chelonian species are rapidly declining due to the international trade of live terrapins for pets, food and medicine, and also the local consumption of eggs and meat, habitat loss and improper coastal and estuarine development (Sharma et al., 1995; CITES, 2006).

PaintedCOPYRIGHT terrapins are aquatic estuarine turtles whereby juveniles can be found in the freshwater areas of rivers while the adults are able to tolerate higher amounts of salinity and can also be found in estuarine areas (Ernst and Barbour, 1989; IUCN, 2000; Bonin et al., 2006). The adults are mainly herbivorous, feeding on leaves and fruits such as fruits of the mangrove tree (Sonneratia sp.) while the juveniles are omnivorous. (Ernst © and Barbour, 1989; Sharma et al., 1995). Adults have marked sexual dimorphism. The female is generally larger with an olive coloured head. The head of the non-breeding male is charcoal grey, which turns white with a red dorsal streak during the breeding

season (Bonin et al., 2006), hence the name Painted terrapin. This beautiful colouration causes them to be a target for the illegal pet trade. The hatchlings and juveniles are monomorphic which makes them impossible to be sexed by appearance alone. Breeding is seasonal, and females will swim out to the estuary and into the sea, to lay their eggs on the sandy beaches (CITES, 2006).

As the nesting sites for the terrapins are the sandy beaches, the eggs have higher risk of being poached or succumb to predation. Thus, to increase the chances of survival, hatcheries have been set up at the major nesting sites. In Malaysia, these hatcheries can be found in Pahang, Terengganu and Malacca, which were set up by the State Fisheries Department (Sharma et al., 1995). The terrapin eggs are collected and re-buried in a safer and enclosed environment (hatchery) for ex situ incubation, away from predators and poachers. However, this method has caused human manipulation of the eggs and its environment. Manipulation of these eggs leads to changes in the incubation temperature which influences the gender of the hatchlings (temperature dependent sex determination), and may lead to a bias in the sex ratio. UPM

One method to increase the population of these terrapins is through captive breeding. However, to date there has not been a successful ex situ conservation breeding program for B. borneoensis (CITES, 2006). This could be due to many factors such as physical health, hormones, environment and nutrition which may not be optimum in captive conditions. Therefore, it is important to conduct comparative health evaluation and reproductive profiling between the captive and wild painted terrapins to determine the causative factors that may prevent them from breeding in captivity.

The present study was therefore undertaken to provide the much needed information on the health and reproductive biology of these critically endangered chelonians inhabiting Sungai Linggi Melaka, Peninsular Malaysia, and also the captive population housed at a zoological facility in Melaka. It is envisaged that the data obtained will assist in the protection and conservation of this turtle species in the wild and also assist in the management of captive individuals in zoological facilities.

The specific objectives of this study are:

1. To evaluate the health (morphological abnormalities, haematology, clinical biochemistry, and parasite burden) of wild and captive Batagur borneoensis in Melaka, Peninsular Malaysia.

2. To profile the temporal reproductive status (breeding colouration, gravid status, COPYRIGHTand reproductive hormone profiles) of wild and captive Batagur borneoensis in Melaka, Peninsular Malaysia.

REFERENCES

Asian Turtle Trade Working Group. (2000). Batagur borneoensis. In: IUCN 2013. IUCN Red List of Threatened Species. Version 2013.2. Retrieved December 24, 2013, from www.iucnredlist.org Baylis, H. (1933). Nematodes from Malayan reptiles. Annals and Magazine of Natural History, 11(10), 615–633. Berry, J. K. (1984). Redescription of Falcaustra duyagi (Tubanguil & Villaamil, 1933) Freitas & Lent, 1941(Nematode: Cosmocercoidea) from Cuora amboinensis (Testudinidae) of Malaysia. Can. J. Zool, 62(9), 1685–1688. Bonin, F., Devaux, B., & Dupre, A. (2006). Geoemydidae. In Turtles of the World. London, UK: A&C Black Publishers. UPM Boyer, T. H., & Boyer, D. M. (2006). Turtle, Tortoises and Terrapins. In D. R. Mader (Ed.), Reptile Medicine and Surgery (Second edi, pp. 85–87). Elsevier Inc. Braun-Mc, J., Epperly, S. P., Owens, D. W., Avens, L., Williams, E., & Harms, C. A. (2007). Seasonal reliability of testosterone radioimmunoassay (RIA) for predicting sex ratio of juvenile Loggerhead (Caretta caretta) turtles. Herpetologica, 63(3), 275–284. Brooks, D. R., & Palmieri, J. R. (1978). Pronocephalid Trematodes from a Malaysian turtle including a new species of Renigonius Mehra, 1939. In Proceedings of the Helminthological Society of Washington (pp. 34–36). Brooks, D. R., & Palmieri, J. R. (1979). Neopronocephalus orientalis sp. n. (Digenea: Pronocephalidae) and Sirhapalum elongatum Rohde, Lee, and Lim, 1968 (Digenea: Spirorchiidae) from Cuora amboinensis (Daudin) in Malaysia. In Proceedings of the Helminthological Society of Washington (pp. 55–57). Brooks, D. R., & Sullivan, J. J. (1981). Hapalorhynchus rugatus sp.nov. (Digena: Spirochiidae) from Malaysian freshwater turtle. Can. J. Zool, 59, 1335–1338. Brophy, T. R. (2006). Allometry and sexual dimorphism in the Snail- eating turtle Malayemys macrocephala from the Chao Phraya River Basin of Central Thailand. Chelonian Conservation and Biology, 5(1), 159–165. Bunkley-Williams, L., Williams, E. H., Horrocks, J. A., Horta, H. C., Mignucci- Giannoni, A. A., & Poponi, A. C. (2008). New leeches and diseases for the Hawksbill sea turtle and the West Indies. Comparative Parasitology, 75(2), 263– COPYRIGHT270. https://doi.org/10.1654/4252.1 Bursey, C. R., & Kinsella, J. M. (2003). Falcaustra greineri n. sp. (Nematoda: Kathlaniidae) from Orlitia borneensis (Testudines: Emydidae). Journal of © Parasitology, 89(5), 961–964.

109

Callard, I. P., Lance, V., Salhanick, A. R., & Barad, D. (1978). The annual ovarian cycle of Chrysemys picta: Correlated changes in plasma steroids and parameters of vitellogenesis. General and Comparative Endocrinology, 35, 245–257. Campbell, T. W. (2006a). Clinical chemistry of reptiles. In M. A. Thrall (Ed.), Veterinary Hematology and Clinical Chemistry (pp. 493–498). Iowa, USA: Blackwell Publishing. Campbell, T. W. (2006b). Clinical pathology of reptiles. In D. R. Mader (Ed.), Reptile Medicine and Surgery (Second). Missouri, USA: Elsevier Saunders. Campbell, T. W. (2006c). Haematology of reptiles. In M. A. Thrall (Ed.), Veterinary Hematology and Clinical Chemistry (pp. 259–274). Iowa, USA: Blackwell Publishing. Campbell, T. W. (2014). Clinical pathology. In D. R. Mader & S. J. Divers (Eds.), Current Therapy in Reptile Medicine and Surgery (pp. 70–89). Missouri, USA: Elsevier Saunders. UPM Casal, A. B., Camacho, M., López-Jurado, L. F., Juste, C., & Orós, J. (2009). Comparative study of hematologic and plasma biochemical variables in Eastern Atlantic juvenile and adult nesting loggerhead sea turtles (Caretta caretta). Veterinary Clinical Pathology, 38(2), 213–218. Casares, M., Rübel, A., & Honegger, R. E. (1997). Observations on the female reproductive cycle of captive Giant tortoises (Geochelone spp .) using ultrasound scanning. Journal of Zoo and Wildlife Medicine, 28(3), 267–273. Cayot, L., & Morillo, G. (1997). Rearing and repatriation of Galápagos tortoises: Geochelone nigra hoodensis, a case study. In J. Van Abbema (Ed.), Proceedings: Conservation, Restoration, and Management of Tortoises and Turtles—An International Conference (pp. 178–183). New York Turtle and Tortoise Society, New York: State University of New York, Purchase. Chabaud, A. G. (1978). No.6. Keys to genera of Superfamilies Cosmocercoidea, Seuratoidea, Heterakoidea and Subuluroidea. In R. C. Anderson, A. G. Chabaud, & S. Willmott (Eds.), Chi keys to the nematode parasites of veterbrates (pp. 9– 17). England: Commonwealth Agricultural Bureaux. Chansue, N., Sailasuta, A., Tangtrongpiros, J., & Wangnaitham, S. (2011). Hematology and clinical chemistry of adult yellow-headed temple turtles (Hieremys annandalii) in Thailand. Veterinary Clinical Pathology, 2, 174–184. https://doi.org/10.1111/j.1939-165X.2011.00306.x Chen, T.-Y., Lee, Y.-T., & Chi, C.-H. (2011). Observation of Reproductive Cycle of Female Yellow-Margined Box Turtle (Cuora flavomarginata) Using Radiography COPYRIGHTand Ultrasonography. Zoo Biology, 30, 689–698. https://doi.org/10.1002/zoo.20376 Cheng-Tsung, T., & I-Jiunn, C. (2013). Two new Records of marine Ozobranchid leeches (Oligochaete : Ozobranchidae ) in Taiwan. Comparative Parasitology, © 80(1), 105–109. Christopher, M. . (1999). Physical and biochemical abnormalities associated with

110

prolonged entrapment in a desert tortoise. Journal of Wildlife Diseases, 35, 361– 366. Combes, C., & Rohde, K. (1979). Polystomoides platynotae n.sp. (Monogenea, Polystomatidae), parasite du chélonien d’eau douce Notochelys platynotae (Gray, 1834) en Malaisie. Vie Milieu, 28–29, 69–75. Congdon, J. D., Breitenbach, G. L., van Loben Sels, R. C., & Tinkle, D. W. (1987). Reproduction and nesting ecology of snapping turtles (Chelydra serpentina) in Southeastern Michigan. Herpetologica, 43(1), 39–54. Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). (2006). Callagur borneoensis. Lima, Peru. Crews, D., Godwin, J., Hartman, V., Grammer, M., Prediger, E. A., & Sheppherd, R. (1996). Intrahypothalamic implantation of progesterone in castrated male whiptail lizards (Cnemidophorus inornatus) elicits courtship and copulatory behavior and affects androgen receptor and progesterone receptor-mRNA expressionUPM in the brain. The Journal of Neuroscience, 16(22), 7347–7352. Dawes, B. (1941). On Multicotyle purvisi n.g., n.sp., an aspidogastrid trematode from the river turtle, Siebenrockiella crassicollis in Malaya. Journal of Parasitology, 33(3), 300–305. Dickinson, V. M., Jarchow, J. L., & Trueblood, M. H. (2002). Hematology and plasma biochemistry reference range values for free-ranging desert tortoises in Arizona. Journal of Wildlife Diseases, 38(1), 143–153. https://doi.org/10.7589/0090-3558- 38.1.143 Divers, S. J. (1997). A clinician’s approach to liver disease in tortoises. In Proceedings of Association of Reptilian and Amphibian Veterinarians (pp. 75–79). Divers, S. J. (2010). Reptile diagnostic endoscopy and endosurgery. The Veterinary Clinics of North America. Exotic Animal Practice, 13(2), 217–42. https://doi.org/10.1016/j.cvex.2010.01.004 du Preez, L. H., & Lim, L. H. S. (2000). Neopolystoma liewi sp. n. (Monogenea : Polystomatidae) from the eye of the Malayan box turtle (Cuora amboinensis). Folia Parasitologica, 47, 11–16. Duguy, R. (1970). Number of blood cells and their variation. In C. Gans & T. Parson (Eds.), Biology of the reptilia (Volume 3, pp. 93–110). London, UK: Academy Press. E.Balasingam. (1962). Some helminth parasites of Malayan reptiles. University of Singapore. Ernst,COPYRIGHT C., & Barbour, R. (1989). Turtles of the World. Washington, D.C. Famelli, S., Adriano, L. R., Pinheiro, S. C. P., Souza, F. L., & Bertoluci, J. (2014). Reproductive biology of the freshwater turtle Hydromedusa maximiliani (Chelidae) from Southeastern Brazil. Chelonian Conservation and Biology, 13(1), © 81–88.

111

Fischthal, J. H., & Kuntz, R. E. (1965). Digenetic trematodes of amphibians and reptiles from North Borneo (Malaysia). Proceedings of the Helminthological Society of Washington, 32(2), 124–136. Franklin, C. J. (2007). Suborder Cryptodira. Hidden necked turtles. In Turtles. An extraordinary Natural History 245 million years in making. U.S.A: Voyageur Press. Garces-Restrepo, M. F., Giraldo, A., Carr, J. L., & Brown, L. D. (2013). Turtle ectoparasites from the Pacific coastal region of Colombia. Biota Neotropica, 13(3), 74–79. Geffré, A., Concordet, D., Braun, J., & Trumel, C. (2011). Reference Value Advisor: a new freeware set of macroinstructions to calculate reference intervals with Microsoft Excel. Veterinary Clinical Pathology, 40(1), 107–112. Gerald, K., Eric, G., & Peter, P. (2011). Endoscopic Imaging of Gonads, Sex Ratio and Temperature Dependent Sex Determination in Captive Bred Juvenile BurmeseUPM Star Tortoises Geochelone platynota. Asian Herpetological Research, 2(4), 240– 244. https://doi.org/10.3724/SP.J.1245.2011.00240 Gibbons, J. W., & Greene, J. L. (1979). X-Ray photography: A technique to determine reproductive patterns of freshwater turtles. Herpetologica, 35(1), 86–89. Graham, L., Schwarzenberger, F., Möstl, E., Galama, W., & Savage, A. (2001). Technical report: A versatile enzyme immunoassay for the determination of progestogens in feces and serum. Zoo Biology, 20, 227–236. Greenblatt, R. J., Work, T. M., Balazs, G. H., Sutton, C. A., Casey, R. N., & Casey, J. W. (2004). The Ozobranchus leech is a candidate mechanical vector for the fibropapilloma-associated turtle herpesvirus found latently infecting skin tumors on Hawaiian green turtles (Chelonia mydas). Virology, 321, 101–110. https://doi.org/10.1016/j.virol.2003.12.026 Gregory, L. F., & Schmid, J. R. (2001). Stress responses and sexing of wild Kemp’s Ridley sea turtles (Lepidochelys kempii) in the Northeastern Gulf of Mexico. General and Comparative Endocrinology, 124(1), 66–74. Greiner, E. C., & Mader, D. R. (2006). Parasitology. In D. R. Mader (Ed.), Reptile Medicine and Surgery (pp. 343–363). Missouri, USA: Elsevier Saunders. Guntoro, J. (2012). The body size and some field notes of Painted terrapin (Batagur borneoensis) in district of Aceh Tamiang, Indonesia. Asian Journal of Conservation Biology, 1(2), 74–77. Hall, D. (2011). Tortoises and turtles. In The ultimate guide to snakes and reptiles (pp. COPYRIGHT78–137). UK: Hermes House. Hernandez-divers, S. J. (2014a). Diagnostic endoscopy. In D. Mader & S. J. Hernandez- divers (Eds.), Current Therapy in Reptile Medicine and Surgery (pp. 154–178). Elsevier Saunders. ©

112

Hernandez-divers, S. J. (2014b). Endoscopy-assisted and Endoscopic Surgery. In D. . Mader & S. J. Hernandez-divers (Eds.), Current Therapy in Reptile Medicine and Surgery (pp. 179–196). Elsevier Saunders. Hernandez-divers, S. J., Stahl, S. J., & Farrell, R. (2009). An endoscopic method for identifying sex of hatchling Chinese box turtles and comparison of general versus local anesthesia for coelioscopy. Journal of the American Veterinary Medical Association, 234(6), 800–804. Hidalgo-Vila, J., Díaz-Paniagua, C., Pérez-Santigosa, N., Plaza, A., Camacho, I., & Recio, F. (2007). Hematologic and biochemical reference intervals of free-living Mediterranean pond turtles (Mauremys leprosa). Journal of Wildlife Diseases, 43(4), 798–801. Hofmeyr, M. D. (2004). Egg production in Chersina angulata : An unusual pattern in a Mediterranean climate. Journal of Herpetology, 38(2), 172–179. Huot-Daubremont, C., Bradshaw, S. D., Bradshaw, F. J., Kuchling, G., & Grenot,UPM C. J. (2003). Variation of plasma sex steroid concentrations in wild and captive populations of Hermann’s tortoise (Testudo hermanni hermanni) in Southern France. General and Comparative Endocrinology, 130, 299–307. https://doi.org/10.1016/S0016-6480(02)00623-8 Innis, C. (2008). Clinical parasitology of the Chelonia. In Small animal and exotics. Proceedings of the North American Veterinary Conference, Volume 22 (pp. 1783– 1785). Orlando, Florida, USA. Innis, C. J., & Boyer, T. H. (2002). Chelonian reproductive disorders. Veterinary Clinics of North America: Exotic Animal Practice, 5(3), 555–578. https://doi.org/10.1016/S1094-9194(02)00013-0 Johnson, J. H. (2004). Husbandry and medicine of aquatic reptiles. Seminars in Avian and Exotic Pet Medicine, 13(4), 223–228. https://doi.org/10.1053/j.saep.2004.04.008 Jones, A. (2001). Family Cladorchiidae Fischoeder, 1901. In A. Jones, R. A. Bray, & D. I. Gibson (Eds.), Keys to the Trematoda Volume 2 (pp. 278–294). UK: CABI Publishing. Joseph, V. (2015). Reptile clinical pathology. In Exotics Conference (pp. 155–162). Kegler, B. (2011). Callagur borneoensis. Retrieved January 10, 2017, from http://animaldiversity.org/accounts/Callagur_borneoensis/ Klingenberg, R. J. (2007). Understanding reptile parasites. California, USA: Advanced Vivarium System. Kuchling,COPYRIGHT G. (1989). Assessment of ovarian follicles and oviductal eggs by ultrasound scanning in live freshwater turtles, Chelodina oblonga. Herpetologica, 45(1), 89– 94. Kuchling, G. (1999). The Reproductive Biology of the Chelonian. Germany: Springer- © Verlag Berlin Heidelberg.

113

Kuchling, G. (2006). Endoscopic sex determination in juvenile freshwater turtles, Erymnochelys madagascariensis : Morphology of gonads and accessory ducts. Chelonian Conservation and Biology, 5(1), 67–73. Kuchling, G., & Kitimasak, W. (2009). Short note endoscopic sexing of juvenile softshell turtles, Amyda cartilaginea. The Natural History Journal of Chulalongkorn University, 9(1)(April), 91–93. Legler, J. M. (1993). Morphology and physiology of Chelonia. In C. Glasby, G. J. Ross, & P. Beesley (Eds.), Fauna of Australia Volume 2A (pp. 1–23). AGPS Canberra. Retrieved from https://www.environment.gov.au/system/files/pages/dc11235d- 8b3b-43f7-b991-8429f477a1d4/files/16-fauna-2a-chelonia-morphology.pdf Licht, P. (1982). Endocrine patterns in the reproductive cycle of turtles. Herpetologica, 38(1), 51–61. Litzgus, J. D., & Mousseau, T. A. (2006). Geographic variation in reproduction in a freshwater turtle (Clemmys guttata). Herpetologica, 62(2), 132UPM–140. https://doi.org/10.1655/05-26.1 Loehr, V. J., Henen, B. T., & Hofmeyer, M. D. (2004). Reproduction of the smallest tortoise, the Namaqualand Speckled padloper, Homopus signatus signatus. Herpetologica, 60(4), 444–454. https://doi.org/10.1655/03-59 Mahmoud, I. Y., & Licht, P. (1997). Seasonal changes in gonadal activity and the effects of stress on reproductive hormones in the common Snapping turtle, Chelydra serpentina. General and Comparative Endocrinology, 107, 359–372. McArthur, S., Meyer, J., & Innis, C. J. (2004). Anatomy and physiology. In S. McArthur, R. Wilkinson, & J. Meyer (Eds.), Medicine and surgery of tortoises and turtles (pp. 57–72). Blackwell Publishing. McPherson, R. J., Boots, L. R., MacGregor, R., & Marion, K. R. (1982). Plasma steroids associated with seasonal reproductive changes in a multiclutched freshwater turtle, Sternotherus odoratus. General and Comparative Endocrinology, 48, 440–451. https://doi.org/10.1016/0016-6480(82)90179-4 Mendonca, M. T., & Licht, P. (1986). Seasonal cycles in gonadal activity and plasma gonadotropin in the Musk turtle, Sternotherus odoratus. General and Comparative Endocrinology, 62, 459–469. Metin, K., Başimoǧlu Koca, Y., Kargin Kiral, F., Koca, S., & Türkozan, O. (2008). Blood cell morphology and plasma biochemistry of captive Mauremys caspica (Gmelin, 1774) and Mauremys rivulata (Valenciennes, 1833). Acta Veterinaria Brno, 77(2), 163–174. https://doi.org/10.2754/avb200877020163 Mueller,COPYRIGHT J. M., Sharp, K. R., Zander, K. K., Rakestraw, D. L., Rautenstrauch, K. R., & Lederle, P. E. (1998). Size-specific fecundity of Desert tortoise (Gopherus agassizii). Journal of Herpetology, 32(3), 313–319. Norton, T., & Wyneken, J. (2015). Body condition scoring the sea turtle. Retrieved © January 26, 2018, from https://lafeber.com/vet/body-condition-scoring-the-sea- turtle/

114

Oliveira-Junior, A., Tavares-Dias, M., & Marcon, J. L. (2009). Biochemical and hematological reference ranges for Amazon freshwater turtle, Podocnemis expansa (Reptilia : Pelomedusidae), with morphologic assessment of blood cells. Research in Veterinary Science, 86, 146–151. https://doi.org/10.1016/j.rvsc.2008.05.015 Owens, D. W. (1997). Hormones in the life history of sea turtles. In P. L. Lutz & J. A. Musick (Eds.), The Biology of Sea Turtles (pp. 315–337). CRC Press. Owens, D. W., Hendrickson, J. R., Lance, V., & Callard, I. P. (1978). A technique for determining sex of immature Chelonia mydas using a radioimmunoassay. Herpetologica, 34(3), 270–273. Palmieri, J. R., & Sullivan, J. T. (1977). Stunkardia minuta sp. n. (Trematoda: Paramphistomidae) from the Malayan box tortoises, Cuora amboinensis. Journal of Helminthology, 51, 121–124. Pichelin, S., Cribb, T., & Bona, F. (1998). Glossocercus chelodinae (MacCallumUPM 1921) n comb (Cestode: Dilepididae) from freshwater turtles in Australia and a redefinition of genus Bancofiella johnston. Systemic Parasitology, 39, 165–181. Platt, T. R., Firth, A., & Sharma, R. S. K. (2011). Redescription of Neopolystoma liewi Du Preez and Lim, 2000 (Monogenea: Polystomatidae), from Cuora amboinensis (Testudines: Geomydidae) with notes on specimen preparation. Comparative Parasitology, 78(2), 286–290. Platt, T. R., & Sharma, R. S. K. (2012). Two new species of Hapalorhynchus (Digenea : Spirorchiidae) from freshwater turtles (Testudines: Geoemydidae) in Malaysia. Comparative Parasitology, 79(2), 202–207. Ramsay, E., & Dotson, T. (1995). Tissue and serum enzyme activities in the yellow rat snake (Elaphe obsoleta quadrivittata). Am J Vet Res, 56, 423–428. Rangel-Mendoza, J., Weber, M., Zenteno-Ruiz, C. E., & López-Luna, M. A. (2009). Hematology and serum biochemistry comparison in wild and captive Central American river turtles (Dermatemys mawii ) in Tabasco, Mexico. Research in Veterinary Science, 87, 313–318. https://doi.org/10.1016/j.rvsc.2009.03.006 Raś-noryńska, M., & Sokół, R. (2015). Internal parasites of reptiles. Annals of Parasitology, 61(2), 115–117. Richardson, J. P. M., & Brooks, D. R. (1987). Polystomoidella mayesi n.sp. (Monogenea: Polystomatidae) from the urinary bladder of a Malaysian box turtle, Cuora amboinensis. Can. J. Zool, 65, 1567–1569. Richardson, L. R. (1969). The family Ozobranchidae redefined, and a novel COPYRIGHTOzobrachiform leech from Murray river turtles (Class Hirudinoidea; Order Rhynchobdelliformes). In Proceedings of the Linnean Society of New South Wales (Vol. 94, pp. 62–80). Rivera, S. (2008). Health assessment of the reptilian reproductive tract. Journal of Exotic © Pet Medicine, 17(4), 259–266. https://doi.org/10.1053/j.jepm.2008.07.003

115

Roberts, J. R., Platt, T. R., & Orélis-Ribeiro, Raphael Bullard, S. A. (2016). New genus of blood fluke (Digenea: Schistosomatoidea) from Malaysian freshwater turtles (Geoemydidae) and its phylogenetic position within Schistosomatoidea. Journal of Parasitology, 102(4), 451–462. Rohde, K. (1962a). A new trematode, Diaschistorchis multitesticularis sp. n., from a Malaysian tortoise, Hieremys annandalei (Boulenger). Journal of Parasitology, 48, 296–297. Rohde, K. (1962b). Parorientodiscus magnus n.g, n.s.p., Ein trematode aus dem darm von Cyclemys amboinensis (Daud) in Malaya. Z. F. Parasitenkunde, 21, 457–464. Rohde, K. (1963). Cotylaspis malayensis n.sp., Der zweite vertreter der Aspidogastrea aus Malaya. Z. F. Parasitenkunde, 22, 283–286. Rohde, K. (1963a). Orientodiscus fernandoi n.sp. and O. hendricksoni n.sp. (Trematoda, Paramphistomata) from the intestine of Trionyx spp. in Malaya. Journal of Helminthology, XXXVII(4), 349–358. UPM Rohde, K. (1963b). Polystomoides malayi n.sp. (Monogenea, Polystomatidae) aus der harnblase von Cyclemys amboinensis in Malaya. Z. F. Parasitenkunde, 22, 278– 282. Rohde, K. (1963c). Uber einige Malayische trematoden. Z. F. Parasitenkunde, 22, 268– 277. Rohde, K. (1965). Studies on the genus Polystomoides Ward, 1917 (Monogenea). Description of 4 Malayan species, a key to the known species, and comparison of the subcuticular layer in Polystomoides and some digenetic trematodes. Zool. Jb. Syst. Bd., 92, 345–368. Rohde, K., Lee, S. K., & Lim, H. W. (1968). Ueber drei Malayische trematoden. Annales de Parasitologie Humaine et Comparee, 43, 33–43. Rostal, D. C., Grumbles, J. S., Lance, V. A., & Spotila, J. R. (1994). Non-lethal sexing techniques for hatchling and immature desert tortoises (Gopherus agassizii). Herpetological Monographs, 8, 83–87. Rostal, D. C., Lance, V. A., Grumbles, J. S., & Alberts, A. C. (1994). Seasonal reproductive cycle of Desert tortoise (Gopherus agassizii) in the Eastern Mojave desert. Herpetological Monographs, 8, 72–82. Rostal, D. C., Owens, D. W., Grumbles, J. S., Mackenzie, D. S., & Amoss, M. S. (1998). Seasonal reproductive cycle of the Kemp’s Ridley sea turtle (Lepidochelys kempi). General and Comparative Endocrinology, 109, 232–243. Rostal, D. C., Paladino, F. V, Patterson, R. M., & Spotila, J. R. (1996). Reproductive COPYRIGHTphysiology of nesting Leatherback turtles (Dermochelys coriacea) at Las Baulas National Park, Costa Rica. Chelonian Conservation and Biology, 2(2), 230–236. Rostal, D. C., Robeck, T. R., Owens, D. W., & Kraemer, D. C. (1990). Ultrasound imaging of ovaries and eggs in Kemp’s Ridley sea turtles (Lepidochelys kempi). © Journal of Zoo and Wildlife Medicine, 21(1), 27–35.

116

Sanjeeva Raj, P. (n.d.). A synopsis of the species of the genus Ozobranchus (De Quatrefages 1852) Hirudinea- Annelida. The Journal of the Bombay Natural History Society, 52, 473–480. Sanjeeva Raj, P., & Penner, L. R. (1962). Concerning Ozobranchus branchiatus (Menzies, 1791) (Piscicolidae : Hirudinea) from Florida and Sarawak. Transactions of the American Mircoscopical Society, 81(4), 364–371. Sawyer, R. T., Taylor, A., & Sahat, H. M. (1982). The leeches of Brunei (Annelida: Hirudinea), with a checklist and key to the known and expected freshwater, terrestial and marine leeches of Borneo. Journal of Brunei Museum, 5(2), 168– 201. Schildger, B. (2013). Endoscopy in reptiles and amphibians. In M. J. Murray, B. Schildger, & M. Taylor (Eds.), Endoscopy in Birds, Reptiles, Amphibians and Fish (pp. 31–54). Endo:Press. Sharma, D. S. (1995). Conservation Of The Painted Terrapin Callagur BorneoensisUPM In Peninsular Malaysia. Sharma, D. S. K. (1996). An update on the Painted terrapin (Callagur borneoensis) research and monitoring at Sg. Linggi, Negeri Sembilan/Melaka. Sharma, D. S. K., Mohd.Nasir, A. S., & Giles, A. (1995). Incubation and hatchling emergence of Painted terrapin (Callagur borneoensis) eggs under hatchery conditions at Paka, Terengganu. In V. M.K, I. Zulkifli, & S. Jasni (Eds.), Proceedings of Seventh Veterinary Association Malaysia Scientific Congress (pp. 122–125). Sharma, D. S. K., Zainal-Zahari, Z., & Sharma, R. S. K. (1995). Husbandry of chelonians at Zoo Melaka: Current practises and recommendations for improvement. In V. M.K, I. Zulkifli, & S. Jasni (Eds.), Proceedings of Seventh Veterinary Association Malaysia Scientific Congress (pp. 101–106). Sharma, R. S. K. (2001). Descriptive morphology, morphometry, community structure and spatial distribution of helminth parasites in the Malayan Box Turtle, Cuora amboinensis (Chelonia : Bataguridae) from Peninsular Malaysia. Universiti Putra Malaysia. Sharma, R. S. K., Rigby, M. C., Sumita, S., Sani, R. A., Vidyadaran, M. ., Jasni, S., & Dailey, M. D. (2002). Redescription of Serpinema octorugatum (Baylis, 1933) (Nematoda:Camallanidae) from the Malayan box turtle Cuora amboinensis (Daudin) (Chelonia: Bataguridae). Systemic Parasitology, 53, 19–28. Sharma, R. S. K., Sharma, D. S. K., & Vidyadaran, M. . (1999). Amblyomma geoemydae (Acari: Ixodidae) collected off the Asian Brown tortoise, Manouria emys from COPYRIGHTTerengganu, Malaysia. Tropical Biomedicine, 16, 43–47. Sharma, R. S. K., Vidyadaran, M., & Mohd Azlan, J. (1998). Amblyomma geoemydae (Acari: Ixodidae) parasite of the Spiny Hill Turtle, Heosemys spinosa. Journal © Veterinar Malaysia, 10(2), 85. Silvestre, M. A. (2013). Hepatic lipidosis in reptiles. In Southern European Veterinary Conference. Barcelona, Spain.

117

Skryabin, K. (1962). Keys to the trematodes of animals and man. (H. P. Arai, Ed.). Illinois, USA: University of Illinois Press. Skryabin, K., Shikhobalova, N., & Mozgovoi, A. (1991). Key to parasitic Nematode Volume 2. (K. Skryabin, Ed.). Netherlands: E.J.Brill. Southwell, T., & Kirshner, A. (1937). A description of a new species of amphistome Chiorchis purvisi with notes on the classification of the genera within the group. Ann. Trop. Med. Parsit, 31, 215–244. Sugnaseelan, S. (1999). A comparative study of the parasitic fauna between invasive turtle species, Trachemys scripta elegans and local chelonians from Peninsular Malaysia. Universiti Putra Malaysia. Sullivan, J. J. (1976). Parapleurogonius brevicecum gen. et sp. n. (Trematoda: Pronocephalidae) from a freshwater turtle in Peninsular Malaysia. Southeast Asian J. Trop. Med. Pub. Hlth., 7(4), 540–542. UPM Telford Jr, S. R. (2009). The Hemogregarines. In Hemoparasites of the Reptilia : Color Atlas and Text (pp. 199–259). U.S.A: Taylor & Francis Group. Tkach, V. V., Platt, T. R., & Greiman, S. E. (2012). A new species of Opisthioglyphe (Trematoda: Telorchiidae) from gall bladder of turtles in Malaysia. Journal of Parasitology, 98(4), 863–868. Tristan, T. (2008). Introduction to hematology and biochemistry in sea turtles. In Proceeding of the ACVP / ASVCP (pp. 2–6). Texas, USA. Turtle Conservation Coalition [Rhodin, A.G.J., Walde, A.D., Horne, B.D., Van Dijik, P.P., Blanck, T., and Hudson, R. (EDS. ). (2011). Turtles in Trouble : The World’s 25+ Most Endangered Tortoises and Freshwater Turtles. Retrieved from http://www.iucn-tftsg.org/wp-content/uploads/file/Top 25/Top_25+_Turtles_in_Trouble_2011.pdf Umapathy, G., Deepak, V., Kumar, V., Chandrasekhar, M., & Vasudevan, K. (2015). Endocrine profiling of endangered tropical chelonians using noninvasive fecal steroid analyses. Chelonian Conservation and Biology, 14(1), 108–115. Van Dijk, P. P., Iverson, J. B., Rhodin, A. G. J., Shaffer, H. B., & Bour, R. (2014). Turtles of the World, 7th Edition : Annotated checklist of taxonomy, synonymy, distribution with maps, and conservation status. Conservation Biology of Freshwater Turtles and Tortoises : Chelonian Research Monographs, 5, 329–479. https://doi.org/10.3854/crm.5.000.checklist.v7.2014 Wilkinson, R. (2004). Clinical pathology. In S. McArthur, R. Wilkinson, & J. Meyer (Eds.), Medicine and surgery of tortoises and turtles (pp. 141–162). Blackwell COPYRIGHTPublishing. Wilson, S. C., & Carpenter, J. W. (1996). Endoparasitic diseases of reptiles. Seminars in Avian and Exotic Pet Medicine, 5(2), 64–74. Witt, D. M., Young, L. J., & Crews, D. (1994). Progesterone and sexual behaviour in © males. Psychoneuroendocrinology, 19(5–7), 553–562.

118

Zhang, F., Gu, H., & Li, P. (2011). A review of chelonian hematology. Asian Herpetological Research, 2(1), 12–20. https://doi.org/10.3724/SP.J.1245.2011.00012

UPM

119

BIODATA OF STUDENT

Vishwanee Kolandaiveloo was born on February 19, 1986 in Penang. She attended Sekolah Rendah Kebangsaan Minden Heights, Penang from 1993 to 1998 and St.George’s Girls’ School, Penang from 1999 to 2003. She then further her studies at Universiti Putra Malaysia for Diploma of Animal Health and Husbandry from 2004 to 2006 and did her degree in Doctor of Veterinary Medicine from 2007 to 2012 at the same university. After her studies, she worked as a veterinarian in small animal clinic, Life Veterinary Clinic in Penang from 2012 to 2013. She then moved to Kuala Lumpur and worked at Aquaria KLCC from 2013 to 2014 before furthering her studies in Master of Veterinary Science in Universiti Putra Malaysia. She is currently working as veterinarian in a small animal clinic, Petcare Veterinary Clinic, Melawati.

UPM

120

LIST OF PUBLICATIONS

Vishwanee, K., Sharma, R.S.K, Sugnaseelan, S. (2012). Effects of olfactory enrichment on diurnal activity and stereotypic behaviour of captive Malayan tigers (Panthera tigris jacksoni). 7th Proceedings of the Seminar in Veterinary Sciences, University Putra Malaysia, Selangor. p. 123.

Vishwanee, K., Sharma, R.S.K, Watanabe, M., Dhaliwal, G.K. (2016). Endoscopic gender determination in the critically endangered painted terrapin (Batagur borneoensis). VAM Congress 2016 Proceedings, Kuching, Sarawak. p. 17. UPM Basripuzi, N.H.H.B, Mohd Shahrom, S., NurMahiza, M.I., Sharma, R.S.K, Nurulaini, R., Mehru, N., Khalida, H., Muhammad Syahir, M., Vishwanee, K., Stear, M. The 26th International Conference of the World Association for the Advancement of Veterinary Parasitology, WAAVP 2017, Kuala Lumpur, Malaysia. p. 258.

Vishwanee, K., Richard, M.S., Ruviniyia, K., Syahir-Mustapa, M., Dhaliwal, G.K., Watanabe, M., Tan, M.M., Sharma, R.S.K. (2018). Reproductive profiling in the critically endangered painted terrapin (Batagur borneoensis). The 11 th International Conference of Asian Society of Conservation Medicine (ASCM), Bali, Indonesia. p. 60

121 UPM