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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 By 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. Copyright © Universiti Putra Malaysia UPM COPYRIGHT © Abstract of the thesis presented to the senate of the Universiti Putra Malaysia in fulfillment of the requirement for the degree, Master of Veterinary Science HEALTH EVALUATION, PARASITE DIVERSITY AND REPRODUCTIVE PROFILING IN THE CRITICALLY ENDANGERED PAINTED TERRAPIN (Batagur borneoensis, Schlegel and Muller, 1844) By 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 i 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 COPYRIGHT © ii 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 COPYRIGHT © iv 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