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EPIDEMIOLOGY of EMERGING DISEASES and DISORDERS in CAGE CULTURED Pangasius Spp

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EPIDEMIOLOGY of EMERGING DISEASES and DISORDERS in CAGE CULTURED Pangasius Spp EPIDEMIOLOGY OF EMERGING DISEASES AND DISORDERS IN CAGE CULTURED Pangasius spp. IN PAHANG, MALAYSIA BY NUR SYAKEERA BINTI MAHMUD A thesis submitted in fulfilment of the requirement for the degree of Master of Science (Biotechnology) Kulliyyah of Science International Islamic University Malaysia JULY 2019 ABSTRACT Pangasius hypophthalmus or famously known by local Malaysians as Patin Hitam is one of the most important sources of food in Malaysia. It is widely cultured in the Peninsular Malaysia especially in Pahang due to the fact that it is a popular consumed freshwater fish. Global economic interest in the fish has increased based on increasing demand in USA and Europe countries. However, high mortality rate of the fish due to bacterial and viral infections gave a negative impact towards the interest which needs to be solved. Therefore, bacteria in P. hypophthalmus in Pahang were being focused including risk factors associated to the prevalence of bacteria and virus in P. hypophthalmus. This research was conducted for two cycles (February – September 2016 and January – August 2017) in several different farms in Temerloh and Pekan, Pahang. Bacteria and virus samples were taken from three organs of P. hypophthalmus; kidney, liver and spleen. Physical parameters for water quality were measured using a multi-parameter probe sensor (YSI, USA) and chemical parameters were analyzed with DR900 colorimeter (Hach, USA). Bacteria samples were identified using biochemical test kits, API 20NE and 20E, followed by confirmation of the bacteria using Polymerase Chain Reaction (PCR). Virus samples were identified using conventional PCR. There were several bacteria isolated throughout the culture period. The highest prevalence of bacteria found was Aeromonas hydrophila (63%) followed by Photobacterium damselae (23%), Plesiomonas shigelloides (7%), Pseudomonas luteola (4%) and Pseudomonas fluorescens (3%). There was no virus isolated from these two sites. The susceptible size of Pangasius spp. that was infested by A. hydrophila was 300 – 400 g. Furthermore, risk factors such as ammonia, nitrite, iron, sulfide, temperature, pH, total suspended solid (TSS), dissolved oxygen (DO) and rainfall rate affected the presence of bacteria were different at all sites. However, only ammonia was present throughout the sampling period at all sites. Sulfide, nitrite, iron and ammonia were the most significant parameters that have strong associations with the presence of A. hydrophila. Significantly, these results could contribute to better treatment of fish disease and development of standard operating procedure of future fish culture for early disease prevention. i خﻻصة البحث إن نبات )Pangasius hypophthalmus( املهشور ند اتجممع ب) Patin Hitam( هي واح ة من املصاد الغذائية املوعة يف ماليزيا. وهي تريب بهشكل واس يف ماليزيا وخاصة يف وﻻية باهدج، وهي معروفة وهلا أمهية اﻻقمصادية العاملية ولذلك يزداد الطلب نليوا من الرﻻيات املمح ة اﻷمرييكية وأو وبا. وم ذلك فإن الراقيات العالية ق يسبب اﻻلموابات البكمريية والفريوسية هلاو تأثري سليب نلى الصحة اﻷمر الذي يمطلب البحث .واحلل وبالمايل فإن البكمرييا املرجردة يف Pangasius hypophthalmus يف باهدج جتري د اسموا مبا يف ذلك نرامل اخلطر املرتبطة باﻻنمهشا البكمريي والفريوسي يف Pangasius hypophtalmus. متت هذه المجربة يف دو تني )فرباير- سبمعرب 6102 ويداير- أغسطس 6102( يف اقفاص مساك خمملفة يف Temerloh و Pekan يف وﻻية باهداج. مت أخذ نيدات البكمريية والفريوسات من ثﻻثة انظاء Pangasius hypophthalmus الكلر والكب والطحاي الطحال. مت قياس املعلعات الفيزيائية جلردة املياة باسمخ ام اجوزة اسمهشعا مبسا املعلعة املمع دة ) ,YSI AMERICA( واملعلعات الكيعيائية حتليلوا باسمخ ام ميقاس اﻷلران ,DR900 (HACH (AMERICA. مت حت ي العيدات البكمريية باسمخ ام مجعرنات اخميا الكيعياء احليرية API 20NE و 20E وبالمايل المحقق من البكمرييا باسمخ ام تفانل البلعرة املمسلسل Polymerase Chain Reaction (PCR) ومت حت ي نيدات الفريوسات باسمخ ام PCR المقلي ية. هداك بدمرييا معزولة حﻻل ة فت حت ي العيدات أنلى البكمرييا املرجردة هي (Aeromonas hydrophila (63% تليوا Photobacterium (Pseudomonas luteola (4%) ,Plesiomonas shigelloides (7%) ,damselae (23% و (Pseudomonas fluorescens (3%. مل يمم العثر نلى فريوس من مراق حت ي العيدات. مقاس .Pangasius spp الذي يصاب بسورلة من طريق A. hydrophila هر 400-300 جرام باﻹضافة إىل ذلك نرامل اخلطر مثل اﻷمرنيا والديتيات واحل ي والكربيمي ود اجة احلرا ة ود اجة احلعرضة واملراد الصلبة العالقة (TSS) واﻷكسيجني املذاب (DO) ومع ل املطر يؤثر إىل وصرل البكمرييا ختملف يف مجي أماكن اليت حت د العيدات. وم ذلك ترج اﻷمرنيا فقط طرال فتة اخذ وحت ي العيدات يف مجي املراق . تعمرب الكربيمي ات والدتيت واحل ي واﻷمرنيا من أهم العرامل اليت ترتبط ابمباطا قريا برجرد A. hydrophila بهشكل كبري. ميكن أن تسوم هذه الدمائج يف حتسيس نﻻج اﻷمراض اﻷمساك وتطرير إجراءات المهشغيل القياسية لهشربية اﻷمساك يف املسمقبل للرقاية من اﻷمراض يف وقت مبكر. ii APPROVAL PAGE I certify that I have supervised and read this study and that in my opinion, it conforms to acceptable standards of scholarly presentation and is fully adequate, in scope and quality, as a thesis for the degree of Master of Science (Biotechnology) ………………………………..… Nur Nazifah binti Mansor Supervisor ………………………………..… Ahmed Jalal Khan Chowdhury Co-Supervisor I certify that I have read this study and that in my opinion it conforms to acceptable standards of scholarly presentation and is fully adequate, in scope and quality, as a thesis for the degree of Master of Science (Biotechnology) ………………………………..… Mohd Fuad bin Miskon Internal Examiner ………………………………..… Fahrul Zaman bin Huyop External Examiner This thesis was submitted to the Department of Biotechnology and is accepted as a fulfilment of the requirement for the degree of Master of Science (Biotechnology) ………………………………..… Mardiana binti Mohamad Ashaari Head, Department of Biotechnology This thesis was submitted to the Kulliyyah of Science and is accepted as a fulfilment of the requirement for the degree of Master of Science (Biotechnology) ………………………..………… Shahbudin bin Saad Dean, Kulliyyah of Science iii DECLARATION I hereby declare that this thesis is the result of my own investigations, except where otherwise stated. I also declare that it has not been previously or concurrently submitted as a whole for any other degrees at IIUM or other institutions. Nur Syakeera binti Mahmud Signature ........................................................... Date .................................. iv INTERNATIONAL ISLAMIC UNIVERSITY MALAYSIA DECLARATION OF COPYRIGHT AND AFFIRMATION OF FAIR USE OF UNPUBLISHED RESEARCH EPIDEMIOLOGY OF EMERGING DISEASES AND DISORDERS IN CAGE CULTURED Pangasius spp. IN PAHANG, MALAYSIA I declare that the copyright holder of this thesis are jointly owned by the student and IIUM. Copyright © 2019 Nur Syakeera binti Mahmud and International Islamic University Malaysia. All rights reserved. No part of this unpublished research may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, recording or otherwise without prior written permission of the copyright holder except as provided below 1. Any material contained in or derived from this unpublished research may be used by others in their writing with due acknowledgement. 2. IIUM or its library will have the right to make and transmit copies (print or electronic) for institutional and academic purposes. 3. The IIUM library will have the right to make, store in a retrieval system and supply copies of this unpublished research if requested by other universities and research libraries. By signing this form, I acknowledged that I have read and understand the IIUM Intellectual Property Right and Commercialization policy. Affirmed by Nur Syakeera binti Mahmud ……..…………………….. …………………….. Signature Date v ACKNOWLEDGEMENTS In the name of Allah, the Most Gracious, the Most Merciful. I would like to express my deepest gratitude to the Almighty Allah. Without the blessing of Him I would not have been able to complete this project. I am also would like to thank to my supervisor Dr. Nur Nazifah Mansor and my co-supervisor Prof. Dr. Ahmed Jalal Khan Chowdhury for their sincere guiding and outstanding supervising. Their wide knowledge, experience and untiring encouragement give me strength along the journey of this research. Special credits to all lab assistants especially to Br. Masrul and Br. Yahya for their kind and sincere assistance and being very supportive throughout this project. I am indebted beyond the words to my entire colleague and friends from Biotechnology department especially Nurul Fatihah, Nur Faiqah, Safwan, Hairi, Nik Nadiah, Suphia Amiera, Nur Amalin Nadia for moral support, professional help and indirect involvement during the hard time have greatly contributed to the completion of this project. I am also would like to give a big appreciation to the National Fish Health Research Division staffs especially to Dr Siti Zahrah, Madam Rimatulhana, Mr. Shahidan, Mr. Fahmi, Ms. Hilma for their guidance and help throughout this study. I wish to express my personal appreciation to my parents, Mahmud bin Makka and Rohani binti Lapo, to my little sister Nur Farha Adila, other siblings, to my husband Mohammad Faiz Hizzuan bin Hanapi and to my daughter Wardah Qaisara for their support, advice and love who always there in my hours of need. Last but not least, thank you very much for those who are directly and indirectly lends a hand in making if this project success. May Allah bless all of you. vi TABLE OF CONTENTS Abstract ........................................................................................................................ i Abstract in Arabic
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