enhancing poultry health and production for sustainable ... · p.x. lim and n.m.a.n.m. faiz 73...

Enhancing Poultry Health and Production for Sustainable Poultry Industry

Proceedings of 3rd World’s Poultry Science Association (Malaysia Branch) and

World Veterinary Poultry Association (Malaysia Branch) Scientific Conference 2018

18 – 19 April 2018

Faculty of Veterinary Medicine, Universiti Putra Malaysia &

Kuala Lumpur Convention Centre, Kuala Lumpur, Malaysia

Editors

Lokman Hakim Idris Azlan Che’ Amat Saleha Abdul Aziz Farina Mustaffa Kamal

Hasliza Abu Hassim Tengku Rinalfi Putra Tengku Azizan Mohd Hezmee Mohd Noor Mohd Mokrish Md Ajat

Hafandi Ahmad Mohd Shahrom Salisi Nor Yasmin Abd Rahaman Intan Shameha Abd Razak

Lau Seng Fong Nik Mohd Faiz Nik Mohd Azmi

Jointly organised by

World’s Poultry Science Association (Malaysia Branch)

World Veterinary Poultry Association (Malaysia Branch) Faculty of Veterinary Medicine, Universiti Putra Malaysia

Department of Veterinary Services, Malaysia United Business Media (M) Sdn Bhd

In conjunction with

Livestock Asia Expo & Forum 2018

Proceedings of 3rd

WPSA (Malaysia Branch) and WVPA (Malaysia Branch) Scientific Conference 2018

ii

©Faculty of Veterinary Medicine

Universiti Putra Malaysia 2018

All right reserved. No parts this publication many be reproduce stored in a retrieval system

or transmitted in any form, or any by means, electronic, mechanical, photocopying,

recording or otherwise, without the prior permission of the copyright owner.

ISBN

978-967-960-364-4

Printed by

Syarikat Perniagaan Weng Sing Taman Sri Serdang, 43300 Seri Kembangan, Selangor, Malaysia

03-89487244 email: [email protected]

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CONTENTS Page

Message from The Vice Chancellor of Universiti Putra Malaysia

Prof. Datin Paduka Dato’ Dr. Aini Ideris 2

Message from The President of World’s Poultry Science Association (Malaysia Branch) & Chairman of The Organising Committee Prof. Dr Mohd Hair Bejo

3

Message from The President of World Veterinary Poultry Association (Malaysia Branch) & Co-Chairman of The Organising Committee Prof. Dr Abd Rahman Omar

4

Organising Committee of the 3rd WPSA-WVPA (Malaysia Branch) Scientific Conference 2018

5

Scientific Programme of the 3rd WPSA-WVPA (Malaysia Branch) Scientific Conference 2018

6

Invited Paper OVERVIEW OF MALAYSIA POULTRY INDUSTRY N.C.N. Jeffrey Federation of Livestock Farmers Associations of Malaysia

8

REGULATIONS ON VETERINARY DRUGS USAGE FROM REGULATORY BODIES AND ITS CHALLANGES Z. Marzuki Department of Veterinary Services of Malaysia

9

THE ROLE AND PRUDENT USE OF ANTIBIOTICS IN POULTRY INDUSTRIES IN MALAYSIA A.K. Tee Malaysian Animal Health and Nutrition Industries Association

10

ISSUES AND CHALLENGES IN PREVENTION AND CONTROL OF FOWL ADENOVIRUS OUTBREAKS M. Hair-Bejo and M.S. Norfitriah Faculty of Veterinary Medicine, UPM

11

CONTROL OF HIGHLY PATHOGENIC AVIAN INFLUENZA IN MALAYSIA M.A. Rohaya, K. Suratan and A.R.S. Bahari Department of Veterinary Services of Malaysia

15

GLOBAL COCCIDIOSIS CONTROL: VACCINATION STRATEGIES AND PERSPECTIVES J. Villa and M. Dardi HIPRA Asia & Oceania

16

WASTE AND ODOUR MANAGEMENT IN POULTRY FARMS N. Nurdiana Neu-Bio Sdn Bhd

17

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TECHNOLOGIES IN CLOSED HOUSE SYSTEM K. Low and E.S. Tan AGCO GSI (Malaysia) Sdn Bhd

19

MANAGING GUT HEALTH FOR OPTIMUM PERFORMANCE IN BROILERS 20 L.W.J. Nernberg BIOMIN Asia-Pasific

CURRENT STATUS AND FUTURE PROSPECTS IN THE DIAGNOSIS OF POULTRY DISEASES A.R. Omar Faculty of Veterinary Medicine, UPM

21

THE IMPACT OF CHICKEN ASTROVIRUS TO POULTRY INDUSTRY T.M.L. Jessica CEVA Animal Health Asia Sdn Bhd

23

MODERN TRACE MINERAL NUTRITION - GETTING MORE FROM LESS T. Tiller and S.A. Elliott Alltech Biotechnology Malaysia Sdn Bhd

26

Poster Paper FEEDING LARVAE MEAL IN BROILER DIETS AND ITS EFFECT ON GROWTH PERFORMANCE M.I. Alshelmani, T.C. Loh, K.Y. Kareem, N.R. Abdulla, H.L. Foo, A.N.M. Zamri and N. Shazali

27

MOLECULAR CHARACTERIZATION OF FOWL ADENOVIRUS ISOLATED FROM COMMERCIAL BROILER FARMS IN MALAYSIA S.N. Syazana, S.W. Tan and A.R. Omar

29

HAEMATOLOGICAL AND BIOCHEMICAL ANALYSIS OF BROILER CHICKENS FED CHITIN AND CHITOSAN ISOLATED FROM CRICKET AND SHRIMP E.B. Ibitoye, I.H. Lokman, M.N.M. Hezmee, Y.M. Goh and A.Z.B. Zuki

32

EFFECT OF SUPPLEMENTING DIETARY ESSENTIAL LIMITING AMINO ACIDS ON FEED FORMULATION, DIGESTIBILITY AND GROWTH PERFORMANCE OF BROILER CHICKENS A. Mujahid, M.K.N. Norzahidi, Z. Zainuddin and N.A. Bakar

34

NUTRIENT DIGESTIBILITY AND GROWTH PERFORMANCE OF BROILER CHICKENS FED WITH DIFFERENT LEVELS OF PALM KERNEL MEAL (PKM) M.K.N. Norzahidi, Z. Zainuddin, N.A. Bakar and A. Mujahid

36

ANTIMICROBIAL USE AND RESISTANCE IN COMMERCIAL VILLAGE CHICKENS: A STUDY ON KNOWLEDGE AND PRACTICES AMONG FARMERS M.I. Nur, A.K. Arifah, N.M.A.N.M. Faiz and M. Rozaihan

38

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FACTORS AFFECTING DRINKING WATER VACCINATION INTAKE IN COMMERCIAL POULTRY FARMS M.Y.V. Chong, N.M Faiz and J.E Lee

40

PATHOGENICITY AND IMMUNOGENICITY OF FOWL ADENOVIRUS OF MALAYSIAN ISOLATE ATTENUATED IN CHICKEN EMBRYO LIVER CELLS IN COMMERCIAL BROILER CHICKENS M.S. Norfitriah, M. Hair-Bejo, A.R. Omar, A. Aini and M.I. Nurulfiza

43

CHARACTERIZATION OF THREE FOWL ADENOVIRUS ISOLATES OF MALAYSIA PROPAGATED IN CHICKEN EMBRYO LIVER CELLS C.C. Ugwu, M. Hair-Bejo, M.I. Nurulfiza, A.R. Omar and I. Aini

45

THE STUDY OF MEAT QUALITY IN MALAYSIAN TURKEY N.J.S. Hawa and I.H. Lokman

47

PATHOGENICITY OF SALMONELLA STANLEY ISOLATE IN COMMERCIAL BROILER CHICKENS R. Balqis and M. Hair-Bejo

49

SEQUENCE ANALYSIS OF S1 GENE OF MALAYSIAN INFECTIOUS BRONCHITIS VIRUS VARIANT AND QX-LIKE STRAINS FOLLOWING SERIAL PASSAGE IN SPECIFIC-PATHOGEN-FREE EMBRYONATED CHICKEN EGGS M.I. Ismail, M. Suwaibah, S.W. Tan, M. Hair-Bejo and A.R. Omar

51

DETECTION OF GASTROINTESTINAL PARASITES IN JAPANESE QUAILS (CORTUNIX CORTUNIX JAPONICA) L. Nursakinah, M.N.M. Hezmee, I.H. Lokman and N.M.M. Isa

54

DETECTION OF HAEMOPARASITES IN JAPANESE QUAIL (CORTUNIX CORTUNIX JAPONICA) A.N.S. Shareen, M.N.M. Hezmee, I.H. Lokman and N.M.M. Isa

56

PATHOGENICITY AND PROPAGATION OF UPMT27 FIELD ISOLATE OF MALAYSIA FOWL ADENOVIRUS IN CHICKEN EMBRYO LIVER CELLS A. Salisu, I. Nurulfiza, A.R. Mariatulqabtiah, M. Hair-Bejo, A.R. Omar and I. Aini

58

EFFECTS OF FEEDING DIFFERENT POSTBIOTICS ON GROWTH PERFORMANCE OF BROILERS UNDER HEAT STRESS A.M. Humam, T.C. Loh, H.L. Foo, S.A. Asmara, M.I. Alshelmani and M.M. Noordin

60

A STUDY OF INTERNAL PARASITES IN FREE RANGE TURKEYS IN SIMPANG RENGGAM FARM, JOHOR, MALAYSIA P.A. Shivani and I.H. Lokman

62

UPREGULATION OF CHOLESTEROL STORAGE SIGNALLING GENES IN HEPG2 CELLS SUPPLEMENTED WITH EDIBLE BIRD NEST (EBN) EXTRACT D. Tawai, M.A.M. Noor, F. Salleh, I.S. Razak, R. Mansor, A. Ideris and M. Ajat

64

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FEEDING PAOB AT DIFFERENT LEVEL OF AMINO ACIDS IN THE DIET OF BROILER CHICKENS C.H. Goh, T.C. Loh and H.L. Foo

66

MYCOFLORA AND NATURAL OCCURRANCE MYCOTOXINS IN CORN, SOYBEAN MEAL AND WHEAT POLLARD IN MALAYSIA N. Shazali, T.C. Loh, H.L. Foo, H. Akit and M.H. Kamalludin

68

SALMONELLA ENTERICA SEROVAR ENTERITIDIS IN MEAT PRODUCTS E. Khoo, H.W. Ho, M.N. Norazariyah, R.S.N. Hanani, M. Nafizah, M.A. Noormah, R.S. Nazri, Y. Rosnah, M. Fhitri, R. Roseliza and A.R. Sohayati

70

EPIDEMIOLOGICAL STUDY OF MYCOPLASMA GALLISEPTICUM AND MYCOPLASMA SYNOVIAE: CASES FROM LABORATORIES IN MALAYSIA FROM 2012- 2017 P.X. Lim and N.M.A.N.M. Faiz

73

DETERMINATION OF NUTRIENT COMPOSITION AND PRESENCE OF BACTERIA IN RAW VERSUS BOILED CHICKEN LIVER DIET K.N. Tan, M.A.R.P. Azaziah, H.A. Hassim and S. Khairani-Bejo

77

THE EFFECT OF DIFFERENT PROTEIN LEVELS ON THE PERFORMANCE OF MARDI KAMPUNG (VILLAGE) CHICKENS AT GROWER PHASE N. Samat, J. Hamed, A.A. Ismail Affendee, S. Rasol, M.N. Ismail, R. Ismail and N. Ishak

79

COMPARISON OF GROWTH PERFORMANCES OF BROILERS FED ON DIETS CONTAINING LOCAL AND IMPORTED GRAIN CORN J. Hamed, S. Rasol, A.H. Zainudin, R. Ismail, N. Ishak and N. Samat

81

INCLUSION EFFECT OF SAGO FLOUR IN DIET ON GROWTH PERFORMANCE OF BROILER QUAILS (COTURNIX COTURNIX JAPONICA) M.S Muhammad and M.N. Hassan

83

OBSERVED AND EFFECTIVE NUMBER OF ALLELES ON COMMERCIAL KAMPUNG CHICKENS IN PENINSULAR MALAYSIA I.A. Azlina-Azma and J.M. Panandam

85

DETECTION OF FUNGAL ORGANISMS IN PIGEONS AT SELECTED AREAS IN SELANGOR A.R.N. Nazihah, A. Jalila, Z. Zunita, K. Krishnammah and R. Rabiatuladawiyah

87

PRELIMINARY STUDY ON EFFECT OF FEEDING MILK EXTRACTED COCONUT MEAT ON EGG PRODUCTION OF KHAKI CAMPBELL DUCKS M.N. Azlian, B.M. Norhafifi, A.P.M. Syahmi, A.A. Hakim and N. Samat

89

Acknowledgements 91

Proceeding of 3rd


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MESSAGE FROM THE VICE CHANCELLOR OF UNIVERSITI PUTRA MALAYSIA

Assalamualaikum W.B.T and Salam Sejahtera It gives me great pleasure to extend a very warm welcome to all participants to Universiti Putra Malaysia (UPM), which is among the top 1% of the best universities in the world and fifth best within ASEAN. UPM's 229th position in the QS World University Ranking, and 36th place in the QS Asia University Ranking are the highest ever achieved. These achievements have made UPM the best university in Malaysia, taking into account the history of its establishment. UPM also won the National Intellectual Property (AHIN) Award, the Most Entrepreneurial Public University award, received a 6 Star in SETARA , produced a World Champion; UPM Olympian Cheong Jun Hoon who became the first Malaysian athlete to win gold medal in the 10m platform diving at the 2017 Aquatic World Championship in Budapest, Hungary, and many other awards at national and international levels - a list of achievements too long to mention here. Greetings to all WPSA and WVPA members, participants, distinguished speakers and guests, to the 3rd WPSA-WVPA (Malaysia Branch) Scientific Conference 2018, with the theme “Enhancing Poultry Health and Production for Sustainable Poultry Industry” at the Faculty of Veterinary Medicine, UPM, and at the Kuala Lumpur Convention Centre, in conjunction with Livestock Asia and Asia MEATEC Expo & Forum from 19 to 21 April 2018. I also wish to take this opportunity to welcome all the invited speakers and various experts from universities, research centres and industries. This conference is jointly organized by WPSA, WVPA, UPM and with the support from UBM and various industry partners. It is gratifying to note that the agenda of this 2-day scientific conference covers a wide range of very interesting topics in both diseases and health as well as nutrition and production. In line with the theme of the conference, papers on current and new innovation and effective strategies in controlling diseases, enhancing health and also maximization of production via nutrition and feed technology will be discussed. Additionally, the best practice in management as well as welfare and breeding will also be covered during the conference. I hope all participants will take this opportunity to interact actively with the speakers and experts in deliberating key strategies and practices in addressing the current and future challenges of the poultry industry. This conference will not be a success without the hard work and commitments of various organisers. I would like to express my sincere appreciations to the members of the organizing committee for their dedication and unwavering efforts in making this event a success. I also would like to thank the sponsors for their support and contributions. I wish you all a successful and productive conference and we look forward to seeing you in UPM and KLCC for a memorable scientific conference and experience. I wish everyone all the best! Thank you “WITH KNOWLEDGE, WE SERVE” Professor Datin Paduka Dato’ Dr. Aini Ideris, FASc.

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MESSAGE FROM THE PRESIDENT OF WORLD’S POULTRY SCIENCE ASSOCIATION (MALAYSIA BRANCH) & CHAIRMAN OF THE ORGANISING COMMITTEE

The poultry sector is an integral part of the livestock industry in Peninsular Malaysia. The tremendous growth in the sector has been largely propelled by the private sector. It has evolved into a progressive, organised and developed industry with an annual production of eggs and meat valued between RM 1.78 billion to RM 6.03 billion annually. The ever increasing Malaysian population requires the poultry industry to continue increasing the supply of food. The Malaysian poultry industry is undergoing major challenges both domestically and globally. To continue to provide future food for Malaysians the current poultry industry has to innovate. Thus, it is indeed timely that the WVPA-WPSA Scientific Conference 2018 focus on the theme “Enhancing Poultry Health and Production for Sustainable Poultry Industry” is organised to address some of these issues. On behalf of the World’s Poultry Science Association (WPSA) (Malaysia Branch), I would like to extend a warm welcome to all participants to the 3rd WPSA-WVPA (Malaysia Branch) Scientific Conference 2018. The WPSA (Malaysia Branch) is pleased to jointly organize the 3rd WPSA-WVPA (Malaysia Branch) Scientific Conference this year together with WVPA (World Veterinary Poultry Association-Malaysia Branch), Universiti Putra Malaysia (UPM), Department of Veterinary Services (DVS) and UBM in conjunction with Livestock Asia 2018. The WPSA (Malaysia Branch) is along established and unique organization that advance knowledge and understanding of all aspects of poultry science and the poultry industry. This year the Conference also include workshop is part of the Livestock Asia 2018. The Conference brings together prominent scientists, academicians, students, industry players, policy makers and individuals, who are passionate with poultry health and production to deliberate on current issues related to innovation of the poultry industry to generate new knowledge for a sustainable industry. I am very delighted to note that the response to this year meeting is overwhelming. Let me take this opportunity to record my sincere thanks and appreciation to the joint organisers, invited speakers, presenters, delegates, sponsors and all individuals who have contributed to the success of this Conference and workshop. The organising committee has done an outstanding job in the promotion and preparation of this event. Their hard work will be rewarded with a very successful Conference. I am fully confident that the presentations and discussions during the Conference will open new horizons in Poultry Science. Finally, I wish you all a truly enjoyable and productive time at the Conference and, for our overseas visitors, a wonderful and memorable time in Kuala Lumpur and a safe return home. Professor Dr Mohd Hair Bin Bejo, FASc.

Proceeding of 3rd


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MESSAGE FROM THE PRESIDENT OF WORLD VETERINARY POULTRY ASSOCIATION (MALAYSIA BRANCH) & CO-CHAIRMAN OF THE ORGANISING COMMITTEE

On behalf of the World Veterinary Poultry Association Malaysia, I would like to extend a warm welcome to all participants of the 3rd World’s Poultry Science Association (WPSA) - World Veterinary Poultry Association (WVPA) - (Malaysia Branch) Scientific Conference 2018 at the Faculty of Veterinary Medicine, Universiti Putra Malaysia and at the Kuala Lumpur Convention Centre, in conjunction with Livestock Asia and Asia MEATEC Expo & Forum from 19 to 21 April 2018. Livestock Asia 2018 is not just a show; it is a venue where expertise and business are evenly combined to foster long-term relationship between clients and traders. WPSA together with WVPA (Malaysia Branch) are honoured to be a part of this event by organising a two-day scientific meeting with the theme of “Enhancing Poultry Health and Production for Sustainable Poultry Industry” with the co-organiser, UBM and support from various industry partners. The WVPA is a global partnership, with more than half a decade of existence, whose objective is to organise meetings to share information on diseases and conditions relating to the avian species, to encourage research in this field, to promote the exchange of information and material for study between individuals and organisations who are interested in the poultry species, and to establish and maintain liaison with other bodies with related interests. I am confident that the 3rd WPSA-WPSA Scientific Conference will provide the latest developments in avian health management, disease control program and new innovations in feed technology and production management through the compilation of papers and presentations from poultry scientists, experts and veterinarians from academia, research centres and industries. I would like to express my deepest gratitude and sincere thanks to the organisers namely WPSA, UPM, UBM and also the support from Department of Veterinary Services, various industry partners, speakers, delegates and all individuals who have contributed directly or indirectly and played a crucial part to the success of this Conference. May I extend to you warm greetings on behalf of the WVPA and the rest of the organising committee and we look forward to seeing you in UPM and KLCC for a memorable scientific conference and experience. Professor Dr Abdul Rahman Omar

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3rd WPSA-WVPA (MALAYSIA BRANCH) SCIENTIFIC CONFERENCE 2018

ORGANISING COMMITTEE

Chairman: Prof. Dr Mohd Hair Bejo Co-Chairman: Prof. Dr Abdul Rahman Omar Secretariat: CENTRAS: Assoc. Prof. Dr Zunita Zakaria Assoc. Prof Dr Siti Khairani Bejo Mr Muhammad Akees Abu Mansor Mr Muhammad Nazmi Zulkifli Ms Shazatul Nazha Secretary: Assoc. Prof. Dr Jalila Abu Assistant Secretary: Dr Rahmat SM Sheriff Treasurer: Prof. Dr Siti Suri Arshad Assistant Treasurer: Dr Suriya Kumari Ramiah Scientific & Proceedings: Dr Lokman Hakim Idris (Head) Dr Hasliza Abu Hassim Prof Dr Saleha Abdul Aziz Assoc. Prof. Dr Mohd Hezmee Mohd Noor Dr Hafandi Ahmad Dr Tengku Rinalfi Putra Tengku Azizan Dr Azlan Che’ Amat Dr Nik Mohd Faiz Nik Mohd Azmi Dr Intan Shameha Abd Razak

Sponsorship: Dato’ Dr Tee Ah Kiat (Head) Prof. Dr Siti Suri Arshad Datuk Jeffrey Ng Choon Ngee Dr Phang Yuen Fun Dr Ahmad Mujahid Prof. Dr Loh Teck Chwen Prof. Dr Zulkifli Idrus Workshop: Prof. Dr Mohd Hair Bejo (Head) Dr Nor Azlina Abdul Aziz Dr Norfitriah Mohamed Sohaimi Dr Azlan Che’Amat Dr Mohd Shahrom Salisi Dr Ahmad Afifi Abdul Ghani Mrs Maizatul Akmal Mokhtar Mr Abd Rashid Abdul Rahman Mr Saipuzaman Ali Mr Mohd Adha P Rameli Mr Siva Soorian Ramasamy Mr Azha Sardi Mr Muhammad Zaid Nor Akahbar Technical & Logistics: Mr Mohd Jamil Samad (Head) Mr Saipuzaman Ali Registration: Mr Muhammad Nazmi Zulkifli Ms Nur Alina Jabir Mrs Noor Saidatul Akma Abu Bakar Mr Mohamad Haikal Hafiz Mohd Razali Protocol & Souvenirs: CENTRAS

Dr Farina Mustaffa Kamal Dr Lau Seng Fong Dr Mohd Shahrom Salisi Dr Mohd Mokrish Md Ajat Dr Nor Yasmin Abd. Rahaman

Hospitality: Dr Hasliza Abu Hassim (Head) Dr Farrah Alias Dr Noraini Samat Mr Ahmad Khairul Wafi Osman

Emcee: Dr Mohd Mokrish Md Ajat

Mr Muhammad Afiq Ghazali Mr Mohd Alif Hairullah Abdul Manaf

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SCIENTIFIC PROGRAMME - DAY 1

18 APRIL 2018 - AUDITORIUM, FACULTY OF VETERINARY MEDICINE, UPM

0815 – 0845 REGISTRATION

0845 – 0910 WELCOMING ADDRESS Prof. Dr Mohd Hair Bejo

Chairman of Organising Committee

OPENING CEREMONY Prof. Datin Paduka Dato’ Dr Aini Ideris

Vice Chancellor, UPM

SCIENTIFIC SESSION 1 Chairman: Dato’ Dr Vincent Ng

0910-0930 OVERVIEW OF MALAYSIA POULTRY INDUSTRY Datuk Jeffrey Ng

Federation of Livestock Farmers Associations of Malaysia

0930 – 0950 REGULATIONS ON VETERINARY DRUGS USAGE FROM REGULATORY BODIES AND ITS CHALLANGES

Dr Marzuki Zakaria Department of Veterinary Services of Malaysia

0950 – 1010 THE ROLE AND PRUDENT USE OF ANTIBIOTICS IN POULTRY INDUSTRIES IN MALAYSIA

Dato’ Dr Tee Ah Kiat Malaysian Animal Health and Nutrition Industries Association

1010 – 1040 FORUM SESSION 1

1040 – 1100 REFRESHMENT

SCIENTIFIC SESSION 2 Chairman: Prof. Dr Abdul Rahman Omar

1100 – 1120 ISSUES AND CHALLENGES IN PREVENTION AND CONTROL OF FOWL ADENOVIRUS OUTBREAKS

Prof. Dr Mohd Hair Bejo Faculty of Veterinary Medicine, UPM

1120 – 1140 CONTROL OF HIGHLY PATHOGENIC AVIAN INFLUENZA IN MALAYSIA Dr Rohaya Mohd Ali

Department of Veterinary Services of Malaysia

1140 – 1200 GLOBAL COCCIDIOSIS CONTROL: STRATEGIES AND PERSPECTIVES Dr Jorge Villa Vinas

HIPRA Asia & Oceania


1220 – 1300 POSTER VIEWING AND JUDGING SESSION

1300 – 1400 LUNCH

WORKSHOP: POSTMORTEM AND DIAGNOSIS ON ADENOVIRUS, NEWCASTLE DISEASE AND COCCIDIOSIS INFECTION IN CHICKENS

1430 – 1530 NECROPSY SESSION

1530 – 1730 MICROSCOPY SESSION


Proceeding of 3rd


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SCIENTIFIC PROGRAMME - DAY 2

19 APRIL 2018 - KUALA KUMPUR CONVENTION CENTER, KLCC 0900 – 1330 EXHIBITION AND OPENING CEREMONY OF THE LIVESTOCK ASIA EXPO AND

FORUM 2018

1330 – 1400 REGISTRATION

1400 – 1410 WELCOMING ADDRESS Prof. Dr Mohd Hair Bejo

Chairman of Organising Committee

SCIENTIFIC SESSION 3 Chairman: Prof. Dr Zulkifli Idrus

1410 – 1430 WASTE AND ODOUR MANAGEMENT IN POULTRY FARMS Nurdiana Nordin Neu-Bio Sdn Bhd

1430 – 1450 TECHNOLOGIES IN CLOSED HOUSE SYSTEM Dr Tan Ee Seng

AGCO GSI (Malaysia) Sdn Bhd

1450 – 1510 MANAGING GUT HEALTH FOR OPTIMUM PERFORMANCE IN BROILER Lesley Nernberg

BIOMIN Asia-Pasific



SCIENTIFIC SESSION 4 Chairman: Prof. Dr Mohd Hair Bejo

1600 – 1620 CURRENT STATUS AND FUTURE PROSPECTS IN THE DIAGNOSIS OF POULTRY DISEASES

Prof. Dr Abdul Rahman Omar Faculty of Veterinary Medicine, UPM

1620 – 1640 THE IMPACT OF CHICKEN ASTROVIRUS TO POULTRY INDUSTRY Dr Jessica Lee

CEVA Animal Health Asia Sdn Bhd

1640 – 1700 MODERN TRACE MINERAL NUTRITION- GETTING MORE FROM LESS Tara Tiller

Alltech Biotechnology Malaysia Sdn Bhd


1720 – 1800 POSTER AWARD AND CLOSING CEREMONY

Proceeding of 3rd


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OVERVIEW OF MALAYSIA POULTRY INDUSTRY

N.C.N. JEFFREY Federation of Livestock Farmers Associations of Malaysia

Corresponding author: [email protected] The presentation will first overview of the Malaysia poultry production for layer and broiler. Review the problem facing by the industry in current situation both internally and externally and also will discuss about the direction for the poultry industry and how to stay sustainable for the future.

mailto:[email protected]

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REGULATIONS ON VETERINARY DRUGS USAGE FROM REGULATORY BODIES AND ITS CHALLANGES

Z. MARZUKI Department of Veterinary Services of Malaysia, Putrajaya

Corresponding author: [email protected]

ABSTRACT

Regulation of veterinary products by relevant authorities depends on product classification. Products containing Scheduled Poison (as in First Schedule of Poison Act 1952), Non-Scheduled Poison / OTC, Pesticides for Internal Use and Pesticides for External Use (control of endoparasite) comes under the purview of National Pharmaceutical Regulatory Agency (NPRA), Pharmacy Division of Ministry of Health (MOH). Products containing animal feed and feed additives regulated under the Animal Feed Act 2009, comes under the jurisdiction of Department of Veterinary Services (DVS), Ministry of Agriculture and Agro based Industries (MOA). There are 5 challenges faced by DVS, pertaining to the 1. Antimicrobial usage (AMU) and Antimicrobial Resistance (AMR) the unwanted phenomena as a result of AMU; 2. Various farms practise different livestock biosecurity measures such closed or open house for protecting its assets from infectious diseases and managing risks to prevent infectious diseases entering livestock properties. AMU tends to be used excessively and intensively in open house as it practices outdated farm biosecurity measures. 3. The concept of organic farming is to use AM as little as possible if not none at all as opposed to conventional farming practices to use AM much as required can lead to reduction of AMR in animal and thus reduced the consequence of AMR in human health, but of course organic livestock products tend to cost more and posed dilemmas. 4. Veterinarians are guided by ethics when administering veterinary products in the food animal practice but the temptation from commercial profits of using more AM may lead to disregard of optimal and ethical use of AM. 5. Legislation need to be enforced to prevent unlawful use of AM by unscrupulous persons, but sometimes the laws maybe incomplete or lack of enforcement which creates loopholes that allows unqualified persons to practice unlawfully.


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THE ROLE AND PRUDENT USE OF ANTIBIOTICS IN POULTRY INDUSTRIES IN MALAYSIA

A.K. TEE

Malaysian Animal Health and Nutrition Industries Association Corresponding author: [email protected]

Poultry Industries are the largest livestock sector in Malaysia. In poultry, antibiotics plays important roles at Prevention, Control, Treatment and Antibiotic Growth Promoter (AGP). In February 2018, FAO said the intense food and agriculture production (including poultry production) in Southeast Asia increased the risks for drug-resistant bacterial infections in humans; and expressed the needs of educating farmers about the dangers of using medically important antibiotics to promote growth in food animals. Local authorities must have an effectives Control System whereby animals on markets and slaughterhouse must be able to trace, so in case of breaking the law farmers can found back and punished. Veterinary companies to promote “alternative to antibiotics” products, including enzymes, probiotics, prebiotics, herbal products and others. On the farming side, farmers should improve farm biosecurity level. Taking these AGP’s out will at this moment have a very big impact on our poultry husbandry. The poor management and farm hygiene will result in an a much higher disease problem, less healthy animals and overnight drop in production, which in return will result in high meat prices and import of poultry meat which does not fit in our economy policies to remain self-sufficiency in poultry meat production. Severe diseases in poultry farming could be reduced via the effective farm hygiene, in line with Good Farming Practice, the assistance of the “alternative to antibiotics” products together with the control and punishment systems in place; thus, the GOAL toward the AGP free production of meat can be implemented. The implementation should be done in steps similar in Europe, with allowing still some AGP’s for a couple of years as a transition period.


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ISSUES AND CHALLENGES IN PREVENTION AND CONTROL OF FOWL ADENOVIRUS OUTBREAKS

M. HAIR-BEJO1,2,* AND M.S. NORFITRIAH1

1Faculty of Veterinary Medicine, 2Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor

*Corresponding author: [email protected] Fowl adenovirus (FAdV) is the causative agent of inclusion body hepatitis (IBH) (Morshed et al., 2017), hydropericardium syndrome (HPS) (Mettifogo et al., 2014) and gizzard erosion (Domanska-Blicharz, 2011) which affect broiler, layer and breeder chickens. It is a double stranded DNA virus with non-enveloped structure comprised of three major structural proteins known as hexon, penton and fiber involved in viral replication in host cells. Antigenic determinants for FAdV located in hexon and fiber proteins contain high amino acid variability between serotype and genotype for FAdV classification (Hess et al., 1998). The virus has been grouped into five molecular species designated as letter A to E and was further divided into 12 serotypes (Meulemans et al., 2004); species A (serotype 1), species B (serotype 5), species C (serotypes 4 and 10), species D (serotypes 2, 3, 9 and 11) and species E (serotypes 6, 7, 8a and 8b) with varying pathogenicity (Pallister et al., 1996). Strains with similar serotypes shown almost identical DNA restriction digestion patterns as compared to strains without cross reaction in neutralization tests with no common fragments (Raue and Hess, 1998). FAdV is endemic worldwide among poultry species involving various geographical regions including Europe (Zadravec et al., 2011), South America (Mettifogo et al., 2014), North America (Gomis et al., 2006), South Africa (Maartens et al., 2014), Middle East Asia (Abdul-Aziz and Al-Attar, 1991), East Asia (Ono et al., 2003), South Asia (Ahmad et al., 1989), Southeast Asia (Hair-Bejo, 2005) and Australia (Erny et al., 1991). From the 1970s to 1990s, IBH caused a significant impact in Australia and New Zealand broiler chickens which majority of outbreak due to FAdV serotype 8 (Steer et al., 2011). To date, the isolate of FAdV belong to serotype 8b are highly distributed in various part of the world (Morshed et al., 2017). In Malaysia, FAdV (IBH) outbreak in commercial broiler chickens due to FAdV species E of serotype 8b was first reported in 2005 (Hair-Bejo, 2005) and since then it was reported in several states of Malaysia involving major poultry producing areas (Juliana et al., 2014; Norina et al., 2016). FAdV is the main pathogen for IBH, but IBH outbreaks also has be reported associated with immunosuppressive diseases such as chicken anaemia virus (CAV) infection (Majdi and Hair-Bejo, 2015). IBH is the most common disease caused by all twelve serotypes of FAdV mostly from species D and E (Ojkić et al., 2008b). The disease occurs in susceptible broiler chickens at 3 to 5 weeks old or even in young age at 7-day-old, and as older as 25 to 27 weeks old (Hair-Bejo, 2005). IBH characterised by sudden onset of high mortality, and it may reach 10% and occasionally higher at 30% (Gomis et al., 2006), which peaked after 3 to 4 days post infection and returned to normal on day 5, but occasionally continued for 2 to 3 weeks. Morbidity is relatively low, however, the sick chickens exhibit


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clinical signs of depression, ruffled feather and reduced feed consumption prior to death (Hafez, 2011). Vertical transmission of FAdV by embryonated chicken eggs in breeder flocks resulting disease outbreak in progeny chicks with poor hatchability and chick quality as well as high mortality in young chicks (Junnu et al., 2015). In addition, oral ingestion of infected faeces in chickens triggers horizontal transmission (McFerran and Adair, 2003). The immunosuppressive effect of infectious bursal disease virus (IBDV) and CAV as well as stress factor enhanced the pathogenicity of FAdV in chickens (Rosenberger et al., 1975). Severe depletion of lymphocytes in bursa of Fabricius and thymus caused by high pathogenic strain from FAdV serotype 4 and 8 resulting immune dysfunction in lymphoid organs (Hussain et al., 2012). FAdV infection can be diagnosed based on the history, clinical signs, lesions and detection, isolation and characterization of the agent. The virus can be isolated using embryonated chicken eggs and tissue culture and detected by electron microscopy. It can be identified either by conventional diagnostic techniques or molecular techniques such as conventional PCR, real time PCR, loop-mediated isothermal amplification (LAMP), in-situ hybridization and real-time PCR. An enzyme-linked immunoabsorbent assay (ELISA) test currently used as serological assay for rapid screening of FAdV infection in chicken flocks. Biosecurity measures are fundamental tool to prevent and control FAdV infection. Proper management, disinfection of house and equipment, restriction of entry into the farm and vaccination play an important role in the prevention and control of the disease (Hafez, 2011). It appears that application of live attenuated FAdV vaccine is superior then inactivated vaccine (Mansoor et al., 2011). The autogenous formalin-inactivated vaccine was reported to be useful to control of HPS outbreak in Pakistan (Afzal and Ahmad, 1990), whilst oil emulsion inactivated vaccine was also effective (Roy et al., 1999). Similarly, in Australia, IBH was successfully prevented by administration of live attenuated vaccine in breeder flocks against homogenous serotype 8 FAdV (Steer et al., 2011). However, recently, sporadic outbreak of IBH in broilers was reported in Australia due to existence of another newly strain isolated from FAdV serotype 11 and required single dual-serotype FAdV vaccine to reduce IBH outbreak in the country (Steer et al., 2011). Proper biosecurity and vaccination programme is crucial to be practiced in Malaysia in the prevention and control of FAdV outbreaks. It is interesting to note that day old chicks from the non FAdV vaccinated flocks have high FAdV antibody titre up to 938+651 to 7795 ± 1414 (Chong, 2017; Norfitriah, 2018) suggesting that the hens were exposed or infected with the field strain of FAdV. Serial passages of field FAdV isolates in embryonated chicken eggs and cell lines especially in chicken liver cells resulting attenuation of the virus with the amino acids and nucleotide changes in the hexon and fibre genes (Alemnesh et al., 2009; Norfitriah et al, 2018). These attenuated FAdV isolates is safe and able to induce high antibody both in SPF and commercial chickens and have high potential to be used for the production of live attenuated FAdV vaccine (Norfitriah, 2018). It was reported that day old broiler chickens (hatchery vaccination) vaccinated with either inactivated, live attenuated or the both the inactivated and live attenuated FAdV serotype 8b showed high

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induction of FAdv antibody titre (3797±980, 1777±600 or 3447±2141 respectively) at 14-day post vaccination in the chickens with maternally derived antibody of 938+651. Vaccination booster at 14 days with the inactivated or the inactivated and attenuated further elevated the FAdV to 4302±2234 or 6312±2232, respectively (Chong, 2017). Vaccination of the breeder flocks with FAdV inactivated vaccine serotype 8b at 16 or 18 weeks of age could induce FadV antibody titre (mean (%CV)) ranging from 7276 (40) to 9908 (15) and 10506(10) to 10983 (7) between 25 to 27 weeks of age. In conclusion, proper biosecurity, accurate diagnosis, appropriate vaccine and vaccination programme are essential to ensure a safe and secure of poultry industry posed by FAdV infection. Enhancing of poultry health and production is vital for sustainable poultry industry in Malaysia and globally. REFERENCES Abdul-Aziz, T.A. and Al-Attar, M. (1991). New syndrome in Iraqi chicks. Veterinary Record, 129(12):

272-272. Afzal, M. and Ahmad, I. (1990). Efficacy of an inactivated vaccine against hydropericardium syndrome

in broilers. Veterinary Record, 126: 59-60. Ahmad, I., Afzal, M., Malik, M.I., Hussain, Z. and Hanif, W. (1989). Studies on the disease pattern and

etiology of hydropericardium syndrome (Angara disease) in broiler chickens in Pakistan. Pakistan Journal of Agricultural Research, 10:195-199.

Alemnesh, W., Hair-Bejo, M., Aini, I. and Omar A.R., (2012). Pathogenicity of Fowl adenovirus in specific pathogen free chickene. Journal of Comparative Pathology, 146(2-3):223-229.

Chong, Z.Z. (2017). Pathogenicity and immunogenicity of live attenuated and inactivated Fowl adenovirus in commercial broiler chickens. DVM Final Year Project 2017, Faculty of Veterinary Medicine, Universiti Putra Malaysia.

Domanska-Blicharz, K., Tomczyk, G., Smietanka, K., Kozaczynski, W. and Minta, Z. (2011). Molecular characterization of fowl adenoviruses isolated from chickens with gizzard erosions. Poultry Science, 90(5): 983–989.

Erny, K.M., Barr, D.A. and Fahey, K.J. (1991). Molecular characterization of highly virulent fowl adenoviruses associated with outbreaks of inclusion body hepatitis. Avian Pathology, 20: 597-606.

Gomis, S., Goodhope, R., Ojkić, D. and Wilson, P. (2006). Inclusion body hepatitis as a primary disease in broilers in Saskatchewan, Canada. Avian Diseases, 50(4): 550-555.

Hafez, H.M. (2011). Avian adenovirus infections with special attention to inclusion body hepatitis/hydropericardium syndrome and egg drop syndrome. Pakistan Veterinary Journal, 31: 85-92.

Hair-Bejo, M. (2005). Inclusion body hepatitis in commercial broiler chickens. Jurnal Veterinar Malaysia, 17(1):33-36.

Hess, M., Prusas, C. and Monreal, G. (1998). Growth analysis of adenoviruses isolated from pigeons in chicken cells and serological characterization of the isolates. Avian Pathology, 27: 196-199.

Hussain, I., Mahmood, M.S., Arshad, M.I., Akhtar, M., Mahmood, F. and Rafique, A. (2012). Immune system dysfunction in broiler chickens experimentally inoculated with fowl adenovirus serotype-4 associated with inclusion body hepatitis hydropericardium syndrome. Turkish Journal of Veterinary and Animal Sciences, 36(3): 223-230.

Juliana, M.A., Nurulfiza, I. Hair-Bejo, M., Omar, A.R and I. Aini (2014). Molecular characterization of fowl adenoviruses isolated from inclusion body hepatitis outbreaks in commercial broiler chickens in Malaysia. Pertanika Journal of Tropical Agricultural Science, 37(4):483-497.

Junnu, S., Lertwatcharasarakul, P., Jala, S., Phattanakulanan, S., Monkong, A., Kulprasertsri, S., Thivalai, C., Chakritbudsabong, W., Chaichoun, K. and Songserm, T. (2015). An inactivated vaccine for prevention and control of inclusion body hepatitis in broiler breeders. Thai Journal of Veterinary Medicine, 45(1):55-62.

Maartens, L.H., Joubert, H.W., Aitchison, H., and Venter, E.H. (2014). Inclusion body hepatitis associated with an outbreak of fowl adenovirus type 2 and type 8b in broiler flocks in South Africa. Journal of the South African Veterinary Association, 85: 1-5.

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Majdi, A. and Hair-Bejo, M. (2015). Pathogenicity of Malaysian fowl adenovirus isolates in specific pathogen free chickens. In: 10

th Proc. the Seminar of Veterinary Sciences, 23-27 February 2015,

Faculty of Veterinary Medicine, Universiti Putra Malaysia. pp. 15-20. Mansoor, M.K., Hussain, I., Arshad, M and Muhammad, G. (2011). Preparation and evaluation of

chicken embryo-adapted fowl adenovirus serotype 4 vaccine in broiler chickens. Tropical Animal Health and Production, 43:331-338.

McFerran J.B. and Adair B.M. (2003). Group I Adenovirus Infections. In: Saif Y.M., Fadly A.M., Glisson J.R., McDougald L.R., Nolan L.K. and Swayne D.E (Eds.). Diseases of Poultry, 12

th edition (pp. 214-

227). Blackwell Publishing, Iowa. Mettifogo, E., Nuñez, Luis F.N., Parra, S.H.S., Astolfi-Ferreira, C.S., and Ferreira, A. J. P. (2014). Fowl

adenovirus Group I as a causal agent of inclusion body hepatitis/hydropericardium syndrome (IBH/HPS) outbreak in Brazilian broiler flocks. Pesquisa Veterinária Brasileira, 34(8): 733-737.

Meulemans, G., Couvreur, B., Decaesstecker, M., Boschmans, M. and Berg, T. P. (2004). Phylogenetic analysis of fowl adenoviruses. Avian Pathology, 33(2): 164-170.

Morshed, R., Hosseini, H., Langeroudi, A.G., Fard, M.H.B. and Charkhkar, S. (2017). Fowl adenoviruses D and E cause inclusion body hepatitis outbreaks in broiler and broiler breeder pullet flocks. Avian Diseases, 61(2): 205-210.

Norfitriah, M.S. (2018). Development of live attenuated Fowl adenovirus isolate of Malaysia for future production of vaccine. PhD Thesis 2018, Faculty of Veterinary Medicine, Universiti Putra Malaysia.

Norfitriah, M.S., Hair-Bejo. M., Omar, A.R., Aini. I., and M.I. Nurulfiza (2018). Molecular detection and pathogenicity of fowl adenovirus isolated from disease outbreak In commercial layer farm. International Journal of Agriculture Sciences and Veterinary Medicine, 6(1): 73-84.

Norina, L., Norsharina, A., Nurnadiah, A., Redzuan, I., Ardy, A., and Nor-Ismaliza. (2016). Avian adenovirus isolated from broiler affected with Inclusion body hepatitis. Malaysian Journal of Veterinary Research, 7(2), 121-126.

Ojkić, D., Krell, P.J., Tuboly, T. and Nagy, E. (2008b). Characterization of fowl adenoviruses isolated in Ontario and Quebec, Canada. Canadian Journal of Veterinary Research, 72:236–241.

Ono, M., Okuda Y., Yazawa, S., Shibata I., Sato S. and Okada K. (2003). Outbreaks of adenoviral gizzard erosion in slaughtered broiler chickens in Japan. Veterinary Record, 153:775-779.

Pallister, J., Wright, P.J. and Sheppard, M. (1996). A single gene encoding the fiber is responsible for variations in virulence in the fowl adenoviruses. Journal of Virology, 70(8): 5115-5122.

Raue, R. and Hess, M. (1998). Hexon based PCRs combined with restriction enzyme analysis for rapid detection and differentiation of fowl adenoviruses and egg drop syndrome virus. Journal of Virological Methods, 73(2): 211-217.

Rosenberger, J.K., Klopp, S., Eckroade, R.J. and Krauss, W.C. (1975). The roles of the infectious bursal agent and several avian adenoviruses in the hemorrhagic-aplastic-anemia syndrome and gangrenous dermatitis. Avian Diseases, 19: 717-729.

Roy, P., Koteeswaran, A. and Manickam, R. (1999). Efficacy of an inactivated oil emulsion vaccine against hydropericardium syndrome in broilers. Veterinary Record, 145(16): 458-459.

Steer, P.A., O’Rourke, D. Ghorashi, S.A. and Noormohammadi, A.H. (2011). Application of high-resolution melting curve analysis for typing of fowl adenoviruses in field cases of inclusion body hepatitis. Australian Veterinary Journal, 89:184–192.

Zadravec, M., Slavec, B., Krape, U., Kaján, G.L., Ranik, J., Juntes, P., Cizerl, R.J., Benko, M. and Rojs, O.Z. (2011). Inclusion body hepatitis associated with fowl adenovirus type 8b in broiler flock in Slovenia – a case report. Slovenian Veterinary Research, 48: 107–113.

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CONTROL OF HIGHLY PATHOGENIC AVIAN INFLUENZA IN MALAYSIA

M.A. ROHAYA1,*, K. SURATAN2 AND A.R.S. BAHARI1 1Department of Veterinary Services Malaysia, Putrajaya 2Department of Veterinary Services Kelantan, Malaysia

*Corresponding author: [email protected] Highly Pathogenic Avian influenza (HPAI) is a zoonotic disease and listed as an OIE notifiable disease in Malaysia. The HPAI subtype H5N1 was first detected in Malaysia on 19 August 2004, followed by second and third experiences in 2006 and 2007 respectively. Thereof, Malaysia was declared free from HPAI H5N1 on 2nd September 2007. Malaysia has remained free from HPAI until February 2017. After an absence of almost 10 years, HPAI re-emerged in 2017 and spread in 6 districts in Kelantan state. In all episodes of HPAI, a stamping out policy has been practised and found to be effective in controlling HPAI in Malaysia. Some countries in the world use vaccines to control HPAI, however, in Malaysia vaccines are strictly prohibited. Since the 1st of July 2017, Malaysia has self-declared for freedom from HPAI infection.


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GLOBAL COCCIDIOSIS CONTROL: VACCINATION STRATEGIES AND PERSPECTIVES

J. VILLA* AND M. DARDI Laboratorios Hipra, S.A.

*Corresponding author: [email protected] Coccidiosis is a parasitic disease caused by coccidia: single-celled animals in the subkingdom Protozoa of the phylum Apicomplexa. All the species of coccidia affecting chickens belong to the genus Eimeria and are predominately host-specific. These species are: Eimeria acervulina, E. maxima, E. tenella, E. brunetti, E. necatrix, E. mitis and E. praecox. The infection by coccidia will receive the name of coccidiosis if there are clinical signs or coccidiasis if the infection is light and there are no clinical manifestations. Coccidia is ubiquitous and most infections in wild birds have no serious consequences for the host. Nevertheless, and due to the conditions in which we produce chickens, coccidiosis is considered one of the diseases with higher economic impact in the poultry industry. There are different strategies to control coccidia infection; we will pay special attention to vaccination strategies in short-life and long-life birds.

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WASTE AND ODOUR MANAGEMENT IN POULTRY FARMS

N. NURDIANA Neu-Bio Sdn Bhd, 38, Jalan PJS 1/28, Taman Petaling Utama 3,

46000 Petaling Jaya, Selangor, Malaysia Corresponding author: [email protected]

ABSTRACT

Nongguan Recycle Quick and Harmless Bio-fermentation system providing a one stop solution to tackle the common problems in managing organic wastes in poultry farming including odor, flies, pollution, and bio-security. Through quick and harmless aerobic bio-fermentation process, poultry manure is treated and converted into high quality organic fertilizer. The process involved is an environmental friendly process with air purification unit to treat the unwanted gases (if produced) from aerobic fermentation process. The organic fertilizer produced from the system is proven as pathogen free and marketable where the properties of organic fertilizer comply with Malaysian Standard. Results from series of laboratory analyses show that the total NPK value of organic fertilizer is more than 6% and the average of C/N ratio is 24:1 (results of analysis subjected to properties of organic wastes used as feedstock). Keywords: waste management, organic fertilizer, aerobic fermentation.

INTRODUCTION Nongguan Recycle Quick and Harmless Bio-Fermentation System is specifically designed for various organic wastes treatment to achieve bio-security in recycling organic wastes into economically valuable product with green and environmental friendly processes. Along with Taiwan in-house developed high temperature Thermophilic microbes, Nongguan Recycle bio-fermentation process can eliminate pathogens, bacteria, parasites, viruses, eggs, and odor in producing organic fertilizers. The bio-fermentation process takes place in an enclosed system where the heat energy can be recycled and the system produce minimum odor. With the aid of air purification unit in the system, fermentation gases such as ammonia and hydrogen sulfide, if produced are absorbed in the air purification column before being released to the environment.

MATERIALS AND METHODS Two main materials involved in producing organic fertilizer from poultry farms wastes are the poultry manure and carbon source material such as sawdust. The mix of poultry manure and carbon source materials is essential in order to reduce the moisture content and to adjust the C/N ratio. The moisture content required in the system must be less than 65% with C/N ratio between the ranges of 25:1 to 30:1. There are two main processes involve in the system which are high temperature aerobic fermentation process, and second aerobic fermentation process. The first fermentation process takes place for 8 hours at 70-80°C and followed by second fermentation process for another 5-7 days. Second fermentation can be conducted in open pit with regulated air system, or in patented fully enclosed cabin type system. The organic wastes are converted into fully fermented pathogen free organic fertilizer with nutrients and organic content required in agriculture. The process flow is referred in Figure 1.


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Figure 1: Process flow

RESULTS AND DISCUSSION As the fermentation process involved in the system is aerobic fermentation process, added with the air purification unit, the odor problems in poultry waste management can be greatly reduced. In addition, through quick and harmless bio-fermentation process, farm wastes are converted into pathogen free and high quality organic or bio-organic fertilizer. In order to comply with Malaysian Standards for organic fertilizer, a series of laboratory analyses of organic fertilizer produced by using this system have been carried out. The summary of the major parameters tested is found in Table 1. These results could vary depends on type and properties of organic wastes involved. Table 1: Laboratory analysis of organic fertilizer

No. Parameters Results Units

1. Total NPK content ≥ 6 % 2. C/N ratio 24:1 %

Microbiological Analysis

3. Escherichia coli ND(<10) CFU/g

4. Salmonella In 25 g Absent

5. Pseudomonas aeruginosa ND(<10) CFU/g

6. Staphyloccocus aerus ND(<10) CFU/g

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TECHNOLOGIES IN CLOSED HOUSE SYSTEM

K. LOW AND E.S. TAN AGCO GSI (Malaysia) Sdn Bhd Plot 218, Lorong Perindustrian Bukit Minyak 6,

Kawasan Perindustrian Bukit Minyak, 14100 Simpang Ampat, Penang, Malaysia Corresponding author: [email protected]

INSULATION AND VENTILATION Essentially the discussion on closed house system revolves around climate control, which based on building insulation properties and ventilation with added cooling or heating. Without these critical parts, it would be an analogy of enclosing a large box over the livestock minus the windows and air movement. Hence key technological implementation would focus on insulation value that allows livestock to thrive at an optimum level and ventilation properties that cools bird efficiently without consuming too much utility. STRUCTURAL DESIGN To maximize insulation and ventilation properties, structural design needs to support insulation materials as well as to ensure it is air-tight and that there are no air leakages during ventilation run time. Drop ceiling design is encouraged, preventing radiant heat from transferring directly down to livestock. Radiant heat can cause poor performance and high mortality rate. A column free structure aside from the main support column encourages smooth airflow and minimizing air turbulence which all leads to utility savings in term of electricity and maintenance. Structural design also needs to be designed according to geographical location in consideration of climate, wind load and snow load. A sound structural design is calculated to produce quick payback in investment and one that reduces future cost of maintenance and upkeep. INTERNET OF THINGS (IOT) The closed-house has evolved from just an environmental control building to a production house with the used of IOT; from a simplified Temperature and Humidity Controller to a full function Management Controller. With GSI proprietary intelligent controller, EDGE; growers are now able to track their farm performance over the internet at any time and place. Growers can also run farm analysis, for example feed consumption, growth rate (ADG) to benchmarking. It can be used as a predictive tool to health of flock and predictive maintenance features can signal the grower when critical equipment needs to be inspected. The seamless farm connectivity is made possible with the utilization of IOT powered by EDGE.


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MANAGING GUT HEALTH FOR OPTIMUM PERFORMANCE IN BROILERS

L.W.J. NERNBERG Biomin Singapore Pte Ltd, 3791 Jalan Bukit Merah, #10-13, E-Centre@Redhill,

159471 Singapore Corresponding author: [email protected]

It is becoming more apparent that gut health and animal performance is closely linked and imperative to efficient commercial livestock production. Any factor that affects gut health will undoubtedly influence the animal as a whole and consequently change its nutrient uptake and requirements, resulting in altered performance and profitability. The basis of gut health lies in its integrity, which is a highly complex process encompassing the macro- and micro-structural integrity of the gut, the balance of microflora, the status of gut-associated immune system and the energetic cost of metabolism. There is a growing body of evidence that stressors can have an impact on several components of the intestinal barrier function and may adversely increase epithelial permeability. With the combination of gut function, immunological, physiological and physical barriers, the gastrointestinal tract protects itself as well as the animal from various physical and physiological stressors. Gut integrity is compromised when the conditions are conducive for increased presence of or exposure to these stressors, which expose the animal to a variety of challenges. These could be either overt clinical conditions or subtle sub-clinical conditions. The sub-clinical challenges happen on a day-to-day basis and generally do not exhibit any visible symptoms. Animals channel their nutrients to overcome these challenges through various means such as the activation of immune system, which otherwise would have been used for growth and productive purposes. This loss has been estimated to be 10-12% of the nutrients absorbed at any given time. Therefore, it is imperative to understand that even a small amount of stress could affect the animal’s productivity and that maintaining an integral and healthy gut is the primary step towards efficient production.


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CURRENT STATUS AND FUTURE PROSPECTS IN THE DIAGNOSIS OF POULTRY DISEASES

A.R. OMAR

Institute of Bioscience, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia

Corresponding author: [email protected] Infectious diseases are one of the major causes of economic losses in poultry industries. In many instances, no specific signs are associated with a particular disease. Besides clinical signs and findings from postmortem examinations, diagnostic tools based on serological and molecular detections are used to confirm the causative agents. However, the identification of causative agents and the detection of specific antibody responses in relation to a clinical problem are often complicated by concurrent infections and improper use of vaccines. Currently, the poultry industry is threatened by either more virulent viruses of endemic diseases or by exotic and emerging diseases that can cause major economic losses to this sector. Catastrophic diseases caused by velogenic Newcastle disease virus (NDV) and highly pathogenic avian influenza (HPAI) are not easy to overlook. The real challenge is to confront subclinical, immunosuppression and concurrent infections which act in concert with other factors such as management, environment and nutrition which form a continuous threat to the entire poultry production system. In addition, the repeated outbreaks of diseases caused by variant strains of infectious bronchitis virus (IBV), genotype VII Newcastle disease viruses (NDV) and more virulent viruses of infectious bursal disease virus (IBDV) and Marek’s disease virus (MDV) in well management poultry flocks, have prompted the need to evaluate the underlying factors contributing to the failure of vaccination to provide complete protection against clinical infections and transmission of disease. Meanwhile, diseases associated with fowl adenovirus, reovirus and astrovirus are emerging disease of concern since these viruses are highly resistant to environment and are often associated with a broad spectrum of diseases that are difficult to diagnose and control. More importantly, these viruses are also transmitted vertically affecting breeder chickens. There is no doubt that diagnostic tools, vaccines and vaccination equipment have improved over the years through the use of innovative technology. However, despite these advancements, conventional laboratory diagnosis using serological tests and conventional live and killed vaccines are used extensively in health and disease management of poultry. However, vaccines are not evolution-proof and, in fact, may enhance virus evolution especially in the absence of sterilizing immunity allowing wild-type and variant strains of viruses to transmit through vaccinated chickens hence, complicate the detection of specific agents. As the poultry industry become more intensive, accurate, economical and practical diagnostic tools are important for effective control of disease outbreaks. Hence, the use of polymerase chain reaction (PCR) has been proven of beneficial in diagnosis of specific disease since the assay is rapid with high specificity and sensitivity. The advancements in the use of molecular detection method using real-time PCR approach, automated


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instruments for antibody detection and development of rapid on-site assays for virus antigen detection may have a significant impact in the field of disease prevention and control. Through the use of real-time PCR in vaccine efficacy study, virus shedding can be determined accurately for the detection of blocking immunity following vaccination. Another emerging application of real-time PCR assay is the detection of immunosuppression. Currently, diagnosis of immunosuppression is based on observation of clinical signs, detecting specific diseases by serologic surveillance, isolation, or molecular detection of the virus; or histological examination to detect depletion or degeneration of lymphoid tissues. Molecular-based assays for the detection of cytokines, chemokines, interferons, and other mediators involved in regulating the immune system have been proposed as a general assessment of the immune function. Hence, real-time PCR have been developed to evaluate the effect of immunosuppressive viruses on the immune system, and also following vaccination. However, the detection of specific viral induced immunosuppressive still requires in vivo experiment in susceptible chickens. Through the use of new sequencing technology based on next-generation sequencing (NGS) technology, microarrays and other functional genomic tools rapid progress has been made in elucidating the molecular mechanisms of viral infection in chickens following inoculation with pathogenic and vaccine strains. However, the clinical importance and field application of the results generated from microarray and NGS studies are difficult to analyse due to the complex interactions of various genes and their related pathways. Further study is required in elucidating the importance of functional genomics study of host-pathogen intearction in the identification of novel markers with diagnostic, therapeutic and prophylactic potentials. In conclusion, as the poultry industry expands and globalization of poultry and poultry by-product, much is needed to improve the control and prevention of diseases. The emerging and emerging of diseases especially the transboundary diseases can impact the socioeconomic of a country and global food security. The strengthening of our scientific and technical capacity, especially through innovative technologies and strategies, present useful ways to help meet the current challenges and future needs of the poultry industry.

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THE IMPACT OF CHICKEN ASTROVIRUS TO POULTRY INDUSTRY

T.M.L. JESSICA Ceva Animal Health Asia Sdn. Bhd. 27-2, Oval Damansara, No. 685, Jalan Damansara, 60000 Kuala Lumpur, Malaysia


ABSTRACT

Chicken astrovirus (CAstV) is a relatively recent emerged virus which has been associated with runting and stunting syndrome that causes poor growth of broiler flocks, enteritis and diarrhea; but also severe kidney lesion in young broilers with visceral gouts and “White Chicks” disease in hatchery. CAstV is the second astrovirus (AstV) of chickens that is separate species from Avian Nephritis Virus (ANV). Being a non-enveloped virus, CAstV are extremely stable and resistant to disinfection and cleaning and can persist in poultry houses for long time. Since its emergence, CAstV are distributed worldwide with some recent reports on its impact towards the poultry production. However, different levels of severity were observed depending of many other factors that suggest more thorough investigation; while in some cases their presence is not related with disease signs. Hence, the impact of CAstV towards poultry industry could range from subclinical to heavy losses. Keywords: Chicken astrovirus (CAstV), Avian Nephritis Virus (ANV), White Chicks disease, kidney lesion, visceral gout.

INTRODUCTION Chicken astrovirus (CAstV) is a newly emerging virus recognized as a member of the avian Astroviruses (AstV). Similar to other astroviruses, CAstV is a non-enveloped, small, round viruses typically with 25 – 30nm in diameter with positive sensed single-stranded RNA genome. Based on species of origin and the characteristics of the viral genome, astrovirus that infect chicken were classified under Avastrovirus 2 genus. In this genus, there are two astrovirus species and both are associated with growth problems, enteritis and kidney lesions in young chickens. The first is avian nephritis virus (ANV), isolated from a one week old, normal broiler chick in 1976 (Yamaguchi et al., 1979). Chicken astrovirus was the second astrovirus, separate species from ANV and commonly found in broiler chickens and hens strongly related to diseases in young birds and hatchery disease (Smyth, 2017).

INFECTION Chicken astrovirus (CAstV) infections usually happen at early age within the first week of life. The earlier the chicks are exposed, either transmitted horizontally by the fecal–oral route, or through vertically transmitted (some CAstV strain) from naive in-lay parent birds, and chicks may hatch shedding high levels of CastV. Depending on the strains (pathogenicity could varies widely), viral load (dose) during infection and presence of maternal derived antibodies (MDA) against CAstV, vertically infected chicks may result in worse outcome. Other factors such as presence of other enteric pathogens such as ANV or ubiquitous enteric viruses, fowl adenoviruses and avian orthoreoviruses could co-infection with CAstV. In addition a flock may be infected with more than one strain of CAstV concurrently. CAstV is very resistant to disinfection and cleaning as it is non-enveloped and darkling beetles can act as vectors for CAstV (Rosenberger, 2010). Smyth (2017) also described that newly placed chicks could quickly became horizontally infected even after cleaning and disinfection using proprietory disinfectants.


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3 TYPES OF DISEASE MANIFESTATION Runting and Stunting Syndrome: CAstV has been associated runting stunting syndrome (RSS) enteritis and growth problems in chicken flocks (Canelli et al. and Kang et al., 2012) but so far, CAstV has yet to be the only etiological agent. Kouwenhoven et al. (1978) described that besides runted and stunted, other symptoms such as enteritis and diarrhoea, leg weakness and irregular feathering was observed. However, CAstV could also be present in underperforming and in good performing flocks. It is still complex and unclear the factors that could lead to poor performance (Symth 2017). Kidney disease and visceral gout: Bulbule et al. (2013) reported severe kidney disease of young broiler chicks with outbreaks of visceral gout and up to 40% mortality in India in 2012. Re-inoculation of SPF chicks resulted in extremely high mortality (67.5% - 100%) 5 and 10 days post-infection with severe kidney lesions and visceral gout. In Middle East, similar CAstV strain with India was isolated from broiler kidneys related to kidney disease and visceral gout in 2010 and 2012 by AFBI’s Stormont laboratory in UK. This strain is still currently circulating based on the 2016 surveillance. Symth (2012) reported that other areas such as Europe and USA were also affected. “White Chick” syndrome (WCS): CAstV was also known for WCS in various Scandinavian countries, North America, Poland and Brazil (Smyth et al., 2013, Nuñez et al., 2016, Sajewicz-Krukowska et al., 2016) but also with the “clubbed down” problem. Affected chicks were weak, runted and died at early age; chicks showed RSS like symptoms, kidney and liver lesions, poor development and abnormal feathering. In Poland, a 4%–5% hatchability decrease was observed for a single breeder flock over a 4-week period when a maximum of 1% of chicks were pale and weak (Sajewicz-Krukowska et al., 2016). In Finland, average 29% of dead in shell embryos could be detected in CAstV affected flocks indicating vertical transmission (Smyth et al., 2013) In Canada, Long et. al. 2017, reported that affected broiler breeder flocks had egg production drops from 0% to 15% and hatchability drops from 1.8% to 49.1%. The financial impacts of WCS to affected hatching egg producers and hatcheries averaged US$4,417 and US$1,287 per 10 000 hens, respectively. CONCLUSIONS Chicken astrovirus (CAstV) is a newly emerging virus with more cases been reported. It is believed that the true impact from this virus is still uncertain due to many other factors. As there are no vaccines or other effective measures being reported, poultry producers need to continue strict biosecurity, increase down time between flocks and use of effective disinfectants to reduce and/or prevent the detrimental impact from CAstV . REFERENCES Agri-Food and Biosciences Institute (AFBI), Stormont Laboratory, Stoney Road, Belfast, BT4 3SD, UK Bulbule, N.R.; Mandakhalikar, K.D.; Kapgate, S.S.; Deshmukh, V.V.; Schat, K.A.; Chawak, M.M. (2013).

Role ofchicken astrovirus as a causative agent of gout in commercial broilers in India. Avian Pathol., 42, 464–473.

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Canelli, E.; Cordioli, P.; Barbieri, I.; Catella, C.; Pennelli, D.; Ceruti, R.; Moreno, A.; Lavazza, A. (2012). Astroviruses as causative agents of poultry enteritis: Genetic characterization and longitudinal studies on field conditions. Avian Dis., 56, 173–182.

Kang, K.I.; El-Gazzar, M.; Sellers, H.S.; Dorea, F.; Willliams, S.M.; Kim, T.; Collett, S.; Mundt, E. (2012). Investigation into the aetiology of runting and stunting syndrome in chickens. Avian Pathol., 41, 41–50.

Kathleen E. Long (2017). Economic Impacts of White Chick Syndrome in Ontario, Canada Avian Diseases, 61:402–408.

Kouwenhoven, B.; Vertommen, M.; van Eck, J.H.H. (1978). Runting and leg weakness in broilers; involvement of infectious factors. Vet. Sci. Commun., 2, 253–259.

Nuñez, L.F.N.; Santander Parra, S.H.; Carranza, C.; Astolfi-Ferreira, C.S.; Buim, M.R.; Piantino Ferreira, A.J. (2016). Detection and molecular characterization of chicken astrovirus associated with chicks that have an unusual condition known as “white chicks” in Brazil. Poult. Sci., 95, 1262–1270.

Rosenberger, J. Darkling beetles as vectors for bacterial and viral pathogens found in poultry litter. In Proceedings of the 45th National Meeting on Poultry Health and Processing, Ocean City, MD, USA, 4–6 October 2010.

Sajewicz-Krukowska, J.; Krzysztof, P.; Lisowska, A.; Pikuła, A.; Zenon, M.; Króliczewska, B.; Doma´ nska-Blicharz, K. (2016). Astrovirus-induced “white chicks” condition—Field observation, virus detection and preliminary characterization. Avian Pathol., 45, 2–12.

Smyth, V.J.; Todd, D.; Trudgett, J.; Lee, A.; Welsh, M.D. (2012). Capsid protein sequence diversity of chicken astrovirus. Avian Path., 39, 151–159.

Smyth, V.J.; Kaukonen, E.; Trudgett, J.; Wylie, M.; Jewhurst, H.; Conway, B.; Welsh, M.D.; Todd, D. (2013). Chicken astrovirus detected in hatchability problems associated with “White Chicks”. Vet. Rec., 173, 403–404.

Stacey L. Schultz-Cherry, Astrovirus Infections, Disease of Poultry 13th

edition, 391. Victoria J. Smyth (2017). A Review of the Strain Diversity and Pathogenesis of Chicken Astrovirus.

Viruses, 9, 29 Yamaguchi, S.; Imada, T.; Kawamura, H. (1979). Characterization of a picornavirus isolated from

broiler chicks. Avian Dis., 23, 571–581.

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MODERN TRACE MINERAL NUTRITION – GETTING MORE FROM LESS

T. TILLER AND S.A. ELLIOTT Alltech Biotechnology, Rama 9 Road, Bangkok, Thailand


The Asian poultry industry continues to grow and remains that largest global chicken producing region in the world, according to the Food and Agriculture Organisation (FAO). This continued growth is due to advances in nutrition, genetics and animal management. The use of organic trace minerals and the total replacement of inorganic minerals in animal diets, is a growing trend in global agriculture. Organic minerals consist of molecules associated with proteins and/or amino acids. The protected chemical structure of organic minerals, reduces interactions with other trace minerals or dietary components in the gastrointestinal tract. For the last 10 years, research into organic trace mineral nutrition requirements has been carried out globally. This body of research demonstrates that it’s possible to use organic minerals, at significantly lower levels than inorganic trace minerals are typically formulated without negatively impacting bird performance. This strategy gives benefits in terms of protection of components such as vitamins and enzymes, protection from contamination risks, while reducing mineral excretion into the environment. Developments in trace mineral nutrition and understanding the true requirements of production birds using organic trace mineral sources, has continuously improved in recent years. Organic mineral technologies enable producers to farm healthier animals while advancing productivity.

mailto:[email protected]*

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FEEDING LARVAE MEAL IN BROILER DIETS AND ITS EFFECT ON GROWTH PERFORMANCE

M.I. ALSHELMANI1,2, T.C. LOH1,3*, K.Y. KAREEM3, N.R. ABDULLA4, H.L. FOO5,6, A.N.

M. ZAMRI3 AND N. SHAZALI3

1Institute of Tropical Agriculture, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia; 2Department of Animal Production, Faculty of Agriculture,

University of Benghazi, Benghazi, Libya; 3Department of Animal Science, Faculty of Agriculture; 4Department of Animal Resource, Salahaddin University - Erbil, Iraq;

5Department of Bioprocess Technology, 6Faculty of Biotechnology and Biomolecular Sciences; Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia

*Corresponding author: [email protected]

ABSTRACT

Larvae meal has been reported as a suitable alternative protein source to fish meal and soybean meal. It contained necessary essential amino acids required in poultry feeding. An experiment was conducted to assess the effects of different levels of larvae meal on broiler performance. A total of 216 one-day-old male broiler chicks were raised for 42 days. Birds were randomly allotted into open house cages and 6 chicks per cage. The dietary treatments were supplemented with larvae meal by 0%, 2%, 4%, 6%, 8% and 10% to represent T1, T2, T3, T4, T5 and T6, respectively. The findings showed that body weight, average daily gain and total weight gain for T1 were significantly decreased (P<0.05) compared to T5 and T6. Keywords: larvae meal, broiler, growth performance, poultry industry.

INTRODUCTION In many countries, soybean meal and fish meal are conventional feedstuffs used as the main source of protein for poultry diets. This is due to their high nutritious content (Rezaeipour et al., 2014). Malaysian livestock industry is heavily dependent on imported feedstuffs, including soybean meal and fish meal (Loh, 2004). There are attempts to substitute yellow corn and soybean meal with local feedstuffs in order to decrease the cost of feed, and to achieve the food security in the developing countries (Alshelmani et al., 2017). Hence, it is important to determine alternative sources of protein to replace aforementioned meals. Earth larvae meal is reported as an alternative feedstuff to fish meal (Prayogi, 2011). The experiment was conducted to investigate the effects of different levels of larvae meal on the growth performance. MATERIALS AND METHODS A total of 216 one-day-old male broiler (Cobb500) chicks purchased from a local company and housed for 42 days in battery cages. Birds were randomly allotted into open house cages containing 6 chicks per cage. Water and feed were provided ad libitum. From 0 to 21 and 22 to 42 days of age, starter and finisher diets were offered to the chickens, respectively. The larvae meal was obtained from black soldier fly (Hermetia illucens). All diets were formulated as isocaloric and isonitrogenous. The dietary treatments were supplemented with larvae meal by 0%, 2%, 4%, 6%, 8% and 10% to represent T1, T2, T3, T4, T5 and T6, respectively. Diets were formulated to meet the nutrient requirements of broiler chickens based on NRC (1994). The experimental animals received humane care as outlined and approved by the

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Institutional Animal Care and Use Committee (IACUC), Universiti Putra Malaysia. Individual body weight (BW) was weekly recorded, and feed intake (FI) per cage was measured. Body weight gain (BWG), average daily weight gain (ADG), total weight gain (TWG) and feed conversion ratio (FCR) were calculated. Data were analyzed using General linear model Procedure of SAS (2014). Duncan’s Multiple Range Test was used to compare the significant differences between treatments. Orthogonal polynomial contrasts were used to test the linear or quadratic nature of the response to incremental concentrations of larvae meal. Test of statistical significance was considered at P< 0.05. RESULTS AND DISCUSSION BW, ADG and TWG was linear and quadratic significantly improved (P<0.05) by including larvae meal comparing to the control. T5 (8% larvae meal) had a quadratic significant improvement (P<0.05) in FCR as compared to the control. The results of growth performance are in accordance with many previous findings (Loh et al., 2009; Julendra H et al., 2012; Rezaeipour et al., 2014). REFERENCES Alshelmani, M. I., Loh, T. C., Foo, H. L., Sazili, A. Q., and Lau, W. H. (2017). Effect of solid state

fermentation on nutrient content and ileal amino acids digestibility of palm kernel cake in broiler chickens. Indian Journal of Animal Sciences 87:1135-1140.

Julendra H, Damayanti E, Lusty Istiqomah SN, and MF, K. (2012). The effectiveness of earthlarva meal supplementation as antibiotic growth promoter replacer with different processing method. The 1

st

Poultry International Seminar:146-157. Loh, T. Year. Livestock production and the feed industry in Malaysia. Proc. 2004. FAO Animal

Production and Health Proceedings (FAO). Loh, T. C., Fong, L. Y., Foo, H. L., Thanh, N. T., and Sheikh-Omar, A. R. (2009). Utilisation of Earthworm

Meal in Partial Replacement of Soybean and Fish Meals in Diets of Broilers. Journal of Applied Animal Research 36:29-32.

NRC. (1994). Nutrient Requirements of Poultry (9th Rev. Ed.). National Academy Press, Washington, DC.

Prayogi, H. S. (2011). The effect of earthworm meal supplementation in the diet on quail’s growth performance in attempt to replace the usage of fish meal. International Journal of Poultry Science 10:804-806.

Rezaeipour, V., Aghajan Nejad, O., and Youseftabar Miri, H. (2014). Growth performance, blood metabolites and jejunum morphology of broiler chickens fed diets containing earthworm (Eisenia foetida) meal as a source of protein. International Journal of Advanced Biological and Biomedical Research 2:2483-2494.

SAS. (2014). SAS 9.4 Output Delivery System: User's Guide. SAS institute.

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MOLECULAR CHARACTERIZATION OF FOWL ADENOVIRUS ISOLATED FROM COMMERCIAL BROILER FARMS IN MALAYSIA

S.N. SYAZANA1, S.W. TAN1 AND A.R. OMAR1*

1Laboratory of Vaccines and Immunotherapeutics, Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor. *Corresponding author: [email protected]

ABSTRACT A total of 55 PCR-positive cases of fowl adenovirus (FAdV) were detected from commercial broiler farms in Malaysia between 2016 and 2017. However, sequence analysis of the FAdV is largely unknown. In this study, molecular characterization of three Malaysian pathogenic FAdV isolates namely UPM/FAdV/392/2017, UPM/FAdV/420/2017 and UPM/FAdV/424/2017 was characterized. Virus isolation was carried out by inoculating 10-day-old SPF embryonated eggs with liver homogenate via chorioallantoic membrane (CAM) route. Virus inoculation result showed 100% mortality of embryo within 7 days of incubation with typical lesion associated with FAdV such as thickening of CAM and multifocal liver necrosis. Pooled CAM and liver of the inoculated chicken embryo were processed for viral DNA extraction and followed with PCR to amplify 1219 bp hexon gene of FAdV. Nucleotide sequence analysis of the amplicon revealed high similarity reaching >99% identity where all of the isolates belonged to a single serotype of FAdV-8b, species E. Pairwise distance analysis between the current isolates with previous Malaysian isolates isolated between 2004 and 2008 also revealed high similarities at the nucleotide level (>95% identity). In addition, the result of phylogenetic analysis also consistent with the clustering of Malaysian FAdV isolates into groups FAdV-8b species E. Keywords: Fowl adenovirus, hexon, Malaysia, PCR, phylogenetic analysis.

INTRODUCTION Fowl adenovirus is a non-enveloped double-stranded DNA virus belongs to family Adenoviridae. Currently, the virus can be grouped into 12 different FAdV serotypes (FAdV-1 to -8a and -8b to -11) with each of them assigned into five different species (FAdV A to E) (Hess, 2000). Different FAdV serotypes are known to cause several diseases in chickens where certain serotypes are linked with particular disease conditions (Schachner et al., 2018). Inclusion body hepatitis (IBH) is the most notable FAdV-associated infection in Malaysia, affecting mainly broilers with high levels of mortality. Molecular detection based on DNA analysis using PCR and sequencing has been used as genetic differentiation tools for genotyping of FAdV. Sequencing and phylogenetic analysis of loop 1 (L1) region of hexon gene have been used to identify the different serotypes of FAdV (Raue and Hess, 1998; Maulemans et al., 2004; Rodriguez et al., 2014; Changjing et al., 2016, Zhang et al., 2016). To date, little is known regarding the molecular typing of FAdV strains circulating in Malaysian broiler farms that cause sporadic outbreaks of FAdV-infections. Identification of FAdV serotypes/genotypes that affect commercial poultry in Malaysia is an important step for efficient vaccination strategies in order to control the disease. Therefore, this study aimed to determine molecular typing of FAdV isolated from commercial broiler farms in Malaysia, based on PCR and phylogenetic analysis of hexon gene. MATERIALS AND METHODS Three FAdV isolates designated as UPM/FAdV/392/2017, UPM/FAdV/420/2017 and UPM/FAdV/424/2017 that were isolated from broiler farms in different states of


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Malaysia were considered in this study. Virus isolation was carried out by inoculation

of 10-day-old SPF embryonated chicken eggs with 200l of liver homogenate. At 8 day post inoculation (DPI), pooled CAM and liver of infected embryos were harvested and homogenized up to 10% (w/v) in 0.01 M phosphate-buffered saline (pH 7.4). Viral DNA extraction was carried out using DNeasy Blood and Tissue Kit (Qiagen, Germany) as described by the manufacturer’s instructions. PCR was carried out to amplify 1219 bp of hexon gene fragment using published primer pair H1/H2 (Raue and Hess, 1998) using TopTaq Master Mix Kit (Qiagen, Germany). The amplified products were sent for sequencing (First BASE, Malaysia). Sequence analysis and comparisons were carried out with other FAdV isolates retrieved from NCBI GenBank database. The phylogenetic analysis of the FAdV isolates was constructed using the Neighbor-Joining method based on Tamura 3-parameter model with 1000 bootstrap replicate values in MEGA 7.0 software. RESULTS AND DISCUSSION SPF embryonated chicken eggs inoculated with the isolates showed 100% mortality within 7 days of incubation. The infected embryo showed typical lesion associated with inclusion body hepatitis (IBH) such as thickened CAM and multifocal liver necrosis (data not shown). The hexon gene of all three isolates was successfully amplified by conventional PCR with the expected band of 1219 bp (data not shown). Analysis of hexon nucleotide sequence revealed that all Malaysian FAdV isolates used in this study were genetically clustered in a single group of FAdV-8b species E. Isolate UPM/FAdV/392/2017 shared 99.2% nucleotide identity with UPM/FAdV/420/2017 and UPM/FAdV/424/2017 isolates respectively. Whereas, isolate UPM/FAdV/420/2017, and UPM/FAdV/424/2017 shared 100% nucleotide identity. The highest nucleotide identity was measured between UPM/FAdV/420/2017 and UPM/FAdV/424/2017 isolates with FAdV-8b strain FAV-LNDL-121011-B previously isolated in China at 98.4% similarity. Pairwise distance analysis was also performed to compare the nucleotide differences between isolates used in this study with Malaysian FAdV isolates collected between 2004 and 2008. The current FAdV isolates have a high percentage of nucleotide similarity ranging from 95.1% to 95.5% with the previously isolated Malaysian FAdV isolates UPM04217, UPM08158 and UPM08136. In addition, phylogenetic analysis supports the clustering of Malaysian FAdV isolates in the same branch with other published FAdV-8b strains (Figure 1).

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Figure 1: Phylogenetic tree of Malaysian FAdV isolates based on hexon gene. Bootstrap values were indicated for the major nodes. The published FAdV strains were used as reference strains for comparison purpose. The Malaysian FAdV isolates used in this study (marked with dot) were grouped with other FAdV-8b reference strains.

ACKNOWLEDGEMENT This study was supported by HICOE grant, Ministry of Higher Education, Malaysia. REFERENCES Changjing, L., Haiying, L., Dongdong, W., Jingjing, W., Youming, W., Shouchun, W., … Yanbo, Y. (2016).

Characterization of fowl adenoviruses isolated between 2007 and 2014 in China. Veterinary Microbiology, 197, 62–67.

Hess, M. (2000). Detection and differentiation of avian adenoviruses: A review. Avian Pathology, 29(3), 195-206.

Meulemans, G., Couvreur, B., Decaesstecker, M., Boschmans, M., & van den Berg, T. P. (2004). Phylogenetic analysis of fowl adenoviruses. Avian Pathology, 33, 164-170.

Raue, R. & Hess, M. (1998). Hexon based PCR combined with restriction enzyme analysis for rapid detection and differentiation of fowl adenoviruses and egg drop syndrome virus. Journal of Virological Methods, 73, 211-217.

Rodriguez, J., Koga, Y., Alvarado, A., & Tinoco, R. (2014). Molecular characterization of Peruvian Adenovirus (FAdV) isolates. Advances in Microbiology, 4, 596-603.

Schachner, A., Matos, M., Grafl, B., & Hess, M. (2017). Fowl adenovirus (FAdV) induced diseases and strategies for their control – a review on the current global situation. Avian Pathology, 1–77.

Zhang, T., Jin, Q., Ding, P., Wang, Y., Chai, Y., Li, Y., Zhang, G. (2016). Molecular epidemiology of hydropericardium syndrome outbreak-associated serotype 4 fowl adenovirus isolates in central China. Virology Journal, 13(1), 1-7.

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HAEMATOLOGICAL AND BIOCHEMICAL ANALYSIS OF BROILER CHICKENS FED CHITIN AND CHITOSAN ISOLATED FROM CRICKET AND SHRIMP

E.B. IBITOYE1, 2, I.H. LOKMAN1*, M.N.M. HEZMEE1, Y.M. GOH1 AND A.Z.B. ZUKI1 1Deptartment of Veterinary Preclinical Sciences, Faculty of Veterinary Medicine,

Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. 2Department of Therio & Animal Production, Faculty of Veterinary Medicine, UDUS,

Nigeria. *Corresponding author: [email protected]

ABSTRACT Dietary components can affect haematological and serum biochemical indices which dictates health status. Chitin and chitosan are known feed additives. Due to paucity of information, this study aims at comparing the effect of cricket and shrimp chitin and chitosan on the haematology and serum biochemistry of broilers. 150 days old Cobb broiler chicks were randomly fed one of the five dietary treatments. Birds in treatment: T1 only basal diet (control); T2, T3, T4 and T5 basal diet+0.5g cricket chitin, cricket chitosan, shrimp chitin and shrimp chitosan/kg diet respectively. It was observed that cricket chitin and chitosan did not (p>0.05) affect the haematology and serum biochemistry, but they reduce serum cholesterol (chol) and triglycerides (trigl), hence they may have a beneficial effect on growth and as hypolipidaemic agent. Although cricket chitin appears to be more effective as a hypolipidaemic agent. Also, cricket or shrimp chitin and chitosan did not vary significantly (p>0.05) regarding blood and serum chemistry. Cricket chitin and chitosan, based on this study are potential feed additives and can substitute commercial chitin and chitosan in broiler production. Keywords: Animal health and production, broiler chickens, chitin, chitosan, haematological and serum biochemistry.

INTRODUCTION The dietary application of antibiotics to promote growth in animal has been prohibited in many countries, and this has led to the evaluation of alternative feed additives (European Union, 2018). Chitin and chitosan are known to improve growth performance of animals with hypocholesterolemic potentials (Keser et al., 2012). Blood picture gives information regarding the health status, so alteration in haematological parameters are used to evaluate health status of animals, and can be influenced by nutrition (Afolabi et al., 2011). However, data on the comparative effect of chitin and chitosan on haematological parameters of broiler chickens are scanty. Presently, available reports on haematological, biochemical and hypolipidaemic profiles of broiler are based on crustacean chitin and chitosan. Similar information on insect chitin and chitosan is needed. Therefore, this study compared the effects of dietary cricket and shrimp chitin and chitosan on haematological and serum biochemical profiles of broiler chickens.

MATERIALS AND METHODS A total of 150 days old Cobb broiler chicks were randomly allotted to one among five dietary treatments (T) replicated thrice. T1 fed basal diet alone (Control); T2, T3, T4

and T5 basal diet+0.5g/kg each of cricket chitin, cricket chitosan (Ibitoye et al., 2018), shrimp chitin and shrimp chitosan (Sigma-Aldrich USA) respectively and were fed ad libitum for 42 d. At d 21 and 42, blood was collected for haematological and serum biochemical profiling. Data were subjected to one-way ANOVA with LSD as post-hoc test at 0.05 significant level using SPSS.


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RESULTS AND DISCUSSIONS At d 42, T4 significantly (p<0.05) reduced Hb and WBC of broiler but within normal ranges. This shows that chitin and chitosan exact no adverse effect on the heart, lungs and bone marrow. The spleen and liver functions in hematopoiesis and organ function depends on its size (Shingleton, 2010). Therefore, significant reduction of Hb and WBC in T4 suggested a decrease in the sizes of these organs. In T3 and T5 non-significant (p>0.05) eosinophilia was noted. In agreement with Aathi et al. (2013), T3 and T5 had increased lymphocyte more than in chitin groups. Mild lymphocytosis is seen in excited or frightened animals (Cossio et al., 2012). It is however not clear if feed additives elicit excitement in birds. The AST was above reference intervals. However, method of determination can influence result of serum chemistry. Wet method used in this study, usually give higher values than the dry slide (Andreasen et al., 1996). Also, higher AST recorded in some treatments in our study might be due to corresponding higher Hb (Khan and Elderderya, 2018). These indicate that the integrity of the internal organs was not affected. In this study, T2-T5 had lowered serum chol and trigl relative to the control. This agreed with Wandira et al. (2016) that dietary chitin have no significant effect on total chol and trigl in ducks. This might be due to similar short study duration, which may have minimized changes in lipid fractions (Bahijri et al., 2017). These results contrast those of Hernawan et al. (2017), probably due to difference in animal health and physiological status. Here, dietary chitin and chitosan reduced serum trigl. Synthesis and secretion of trigl by hepatocytes is triggered by insulin (Zammit, 2013). In chickens, glucosamine is absorbed following partial hydrolysis of chitin and chitosan. This glucosamine impairs glucose-induced insulin secretion (Yanagida et al., 2014), consequently lowering hepatic lipogenesis and trigl synthesis in broiler chickens. It was concluded that, there was no significant different between cricket and shrimp chitin and chitosan on their effect on haematological and serum biochemical profile of broiler chickens.

REFERENCES Aathi, K., Ramasubramanian, V., Uthayakumar, V., Munirasu, S., (2013). Effect of Chitosan

Supplemented Diet on Survival, Growth, Hematological, Biochemical and Immunological Responses of Indian Major Carp Labeo Rohita. Int. Res. J. Pharm. 4, 141–147.

European Union, (2018). Register of Feed Additives. pursuant to Regulation (EC) No 1831/2003, Edition

1/. ed. European Union legislation on feed additives. doi:10.2875/48257 Hernawan, E., Adriani, L., Mushawwir, A., Cahyani, C., Darmawan, (2017). Effect of Dietary

Supplementation of Chitosan on Blood Biochemical Profile of Laying Hens. Pakistan J. Nutr. 16, 696–699.

Ibitoye, E.B., Lokman, I.H., Hezmee, M.N.M., Goh, Y.M., Zuki, A.B.Z., Jimoh, A.A., (2018). Extraction and physicochemical characterization of chitin and chitosan isolated from house cricket. Biomed. Mater 13. doi:10.1088/1748-605X/aa9dde

Khan, M.N., Elderderya, A., (2018). Alterations of Haematological Parameters, Haemoglobin and Haematocrit with Chronic Liver Disease Undergoing Hemodialysis in Aljouf Region, Saudi Arabia 7, 1-6.

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EFFECT OF SUPPLEMENTING DIETARY ESSENTIAL LIMITING AMINO ACIDS ON FEED FORMULATION, DIGESTIBILITY AND GROWTH PERFORMANCE OF BROILER

CHICKENS

A. MUJAHID1*, M.K.N. NORZAHIDI1, Z. ZAINUDDIN1 AND N.A. BAKAR1 1Animal Nutrition Technology Centre, Sumitomo Chemical Enviro-Agro Asia Pacific


ABSTRACT

The effect of supplementing limiting synthetic amino acids (SAA, DL-Methionine, L-Lysine and L-Threonine) in broiler feed on least-cost feed formulation, nutrient composition, digestibility, feed cost and growth performance of broiler chickens was studied. Chicks were fed four isocaloric-feeds ad libitum with various protein and amino acid levels. Feed cost, protein and fat contents linearly decreased with SAA addition. Feed without SAA supplementation increased the percentage use of soybean meal and vegetable oil while decreased corn levels and was higher in crude protein, fat, fiber, ash, linoleic acid, arginine, lysine, cystine, threonine, tryptophan, leucine, isoleucine, histidine, phenylalanine, tyrosine, glycine and valine contents. Average body weight, feed consumption, FCR and production efficiency factor (PEF) were similar among different treatments either with or without SAA. Supplementing SAA in broiler feed did not significantly affect production performance and feed efficiency of broiler chickens (P>0.05). SAA during broiler starter phase improved methionine and protein digestibilities and subsequently decreased the excreta nitrogen while supplementing synthetic DL-Methionine in broiler grower feed improved fat digestibility. In conclusion, supplementing DLM improved fat digestibility while SAA supplementation decreased excreta nitrogen, feed protein and fat contents, and cost of broiler feeds without affecting the growth performance and PEF of broiler chickens. Keywords: limiting amino acids, digestibility, excreta nitrogen, least-cost feed formulation, broiler.

INTRODUCTION A better understanding of nutritional requirements of amino acids allows a more precise nutrition, offering the possibility for the formulator to optimize the requirement levels of crude protein by essential amino acids requirements, generating better result and lower costs for the producer (Pesti, 2009). Reducing dietary crude protein is an efficient way to limit nitrogen excretion in broilersit often reduces performance, probably because of an inadequate provision of amino acids (Belloir et al., 2017). Diets are usually formulated using linear least cost software frequently setting a minimum concentration of crude protein in a strategy to have minimum levels of indispensable and dispensable amino acid intake so that performance is not limited (Vieira et al., 2016). This present study aims to determine the effect of supplementing synthetic limiting amino acids (DL-Methionine, L-Lysine and L-Threonine) in broiler feed on feed formulation, feed protein, fat and amino acid levels, digestibility, feed cost and growth performance of broiler chickens. MATERIALS AND METHODS Two hundred unsexed Ross308 broiler chicks were divided into 4 treatment groups with 5 replicates each (10 birds per replicate). Chicks fed with four isocaloric feeds with various protein and amino acid levels that were balanced without or with supplementation of different synthetic amino acids, (SAA). For digestibility experiment (total excreta collection), 90 unsexed Ross308 broiler chickens were used.

mailto:*Corresponding%20author:%[email protected]

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RESULTS AND DISCUSSION Cost of feed (Figure 1), protein and fat contents linearly decreased with addition of SAA. Feed without SAA supplementation increased the percentage use of soybean meal and vegetable oil while decreased the corn levels. Feed without SAA was higher in crude protein, fat, fiber, ash, linoleic acid, arginine, lysine, cystine, threonine, tryptophan, leucine, isoleucine, histidine, phenylalanine, tyrosine, glycine and valine contents. Average body weight, feed consumption, FCR and PEF were similar among groups either with or without SAA (P>0.05, Table 1). SAA during broiler starter phase improved methionine and protein digestibilities and subsequently decreased the excreta nitrogen (Figure 2) while supplementing synthetic DL-Methionine in broiler grower feed improved fat digestibility. In conclusion, supplementing DLM improved fat digestibility while SAA supplementation decreased excreta nitrogen, feed protein and fat contents, and cost of broiler feeds without affecting the growth performance and PEF of broiler chickens. Table 1. Performance of broiler chickens fed without or with synthetic amino acid supplementation

a-b Values within row with different superscripts differ significantly (p<0.05)

REFERENCES Belloir, P., Meda B., Lambert, W., Corrent, E., Juin, H., Lessire, M., Tesseraud, S. (2017). Reducing the

CP content in broiler feed: impact on animal performance, meat quality and nitrogen utilization. Animal, 11 (11):1881-1889.

Pesti, G.M. (2009). Impact of dietary amino acid and crude protein levels in broiler feeds on biological performance. Journal of Applied Poultry Research, 18 (3):477-486.

Vieira, S.L, Stefanello, C. and Cemin, H.S. (2016). Lowering the dietary protein levels by the use of synthetic amino acids and the use of a mono component protease. Animal Feed Science and Technology. 221: 262-266.

Parameter No SAA DLM DLM+Lys DLM+Lys+Thr

Broiler Starter

AW (g) 235.00 ± 1.05b

260.80 ± 4.33a

235.80 ± 2.08b

253.60 ± 6.73a

FC (g) 214.80 ± 3.60 234.40 ± 7.67 229.20 ± 1.07 234.00 ± 12.35

FCR 0.914 ± 0.013 0.901 ± 0.038 0.972 ± 0.012 0.922 ± 0.030

Broiler Grower

AW (g) 995.80 ± 13.93b

1072.00 ± 21.44ab

1057.00 ± 7.16ab

1104.00 ± 34.44a

FC (g) 1289.40 ± 17.86 1394.80 ± 23.41 1349.20 ± 15.79 1400.40 ± 50.76

FCR 1.297 ± 0.034 1.302 ± 0.013 1.276 ± 0.012 1.268 ± 0.015

Broiler Finisher

AW (g) 1956.60 ± 32.60 2038.40 ± 49.51 2055.40 ± 38.03 2059.80 ± 55.83

FC (g) 3133.60 ± 40.51 3263.80 ± 73.28 3176.02 ± 47.66 3312.60 ± 117.56

FCR 1.601 ± 0.035 1.602 ± 0.023 1.546 ± 0.012 1.609 ± 0.041

PEF 337.322 ± 17.852 356.646 ± 13.727 364.478 ± 9.589 344.52 ± 18.393

150

160

170

180

190

200

No SAA DLM+Lys

Pri

ce/1

00

Kg

(RM

)

Figure 1. Ingredient cost of feeds

StarterGrowerFinisher

DLM+Lys+ThrDLM

a

b

abb

0

1

2

3

4

5

6

7

8

No SAA DLM DLM+Lys DLM+Lys+Thr

Nit

rog

en

co

nte

nt

(%)

Figure 2. Excreta nitrogen contents (DMB)

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NUTRIENT DIGESTIBILITY AND GROWTH PERFORMANCE OF BROILER CHICKENS FED WITH DIFFERENT LEVELS OF PALM KERNEL MEAL (PKM)

M.K.N. NORZAHIDI, Z. ZAINUDDIN, N.A. BAKAR AND A. MUJAHID*

Animal Nutrition Technology Centre, Sumitomo Chemical Enviro-Agro Asia Pacific *Corresponding author: [email protected]

ABSTRACT

Palm kernel meal (PKM), a by-product obtained after the extraction of palm kernel oil from the palm kernel seed that is extensively cultivated in tropical countries and it is a potentially valuable feed ingredient for poultry. Different dietary levels of PKM were evaluated in broiler feed for least-cost feed formulation, nutrient contents, digestibility, feed cost and growth performance of broiler chickens. Chicks were fed ad libitum with isocaloric and isonitrogenous diets with various level of PKM (0%, 5% and 10%). The replacement of corn and soya bean meal (SBM) with PKM in least-cost feed formulation subsequently increased vegetable oil ratio in feed. The addition of PKM in broiler feed increased the feed cost up to 3%. No significant differences were found (P>0.05) in average body weight, feed consumption and FCR. Feeding 10% PKM in starter phase resulted in significant decrease (P<0.05) of the digestibility of ash, glutamic acid, methionine, arginine and proline while crude fat and histidine digestibilities were increased as compared to control feed. In conclusion, data indicated that PKM inclusion in broiler feed replaced corn and SBM up to 10% without any statistically significant effect on production performance of broiler chickens; however, it increased the feed cost up to 3%. Keywords: Palm kernel meal, least-cost feed formulation, broiler, growth performance, digestibility.

INTRODUCTION The poultry industry today is highly intensified and has contributed greatly to cheap, abundant meat supply and improved quality of life. High demand for conventional feed ingredients due to competition with humans and other livestock species for feed resources and ever-increasing cost of these ingredients have motivated poultry industry to maximize the use of locally available by-products (Abdollahi et al., 2016). Palm kernel meal (PKM), a by-product obtained after the extraction of palm kernel oil from the seed that is extensively cultivated in tropical countries and it is an interesting feed ingredient for poultry due to its availability (Alshelmani et al., 2016). This study was aimed to investigate growth performance of broiler chickens and nutrient digestibility of feeds with different dietary inclusion levels of palm kernel meal. MATERIALS AND METHODS A total of 135 one-day-old unsexed broiler chicks (Ross308) was raised in closed house system for growth performance (90 chicks) and nutrient digestibility of feeds (45 chicks) with different dietary levels of PKM in broiler chickens. Chicks were individually weighed and randomly allocated into 18 pens (10 chicks per pen for growth trial and 5 chicks per pen for digestible trial). Birds were offered ad libitum isocaloric and isonitrogenous diets containing 0, 5 and 10% PKM at three different phases (Starter, d0-10; Grower, d11-24; Finisher, d25-35). Chicken feed intake, body weight, FCR and digestibilities of PKM were evaluated at day 10, 25, and 35 of age. The digestibility values were calculated from a total excreta collection method. Data were analysed using computer software IBM SPSS Statistics version 23.0. Data were evaluated through ANOVA, and means were compared by Tukey’s test with value P<0.05 considered significant.


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RESULTS AND DISCUSSION Replacement of corn and SBM with PKM in isocaloric and isonitrogenous broiler feed subsequently increased vegetable oil ratio. The addition of PKM (5 and 10%) in broiler feed increased the feed cost up to 3% (Fig. 1). Feeding 10% PKM in starter phase resulted in significant decrease (P<0.05) in the digestibilities of ash, glutamic acid, methionine, arginine and proline while crude fat and histidine digestibilities were significantly increased as compared to control feed. However, digestibilities of crude protein, fiber and other amino acids under study were not affected. During broiler grower phase, feeding 10% PKM showed significantly increased digestibilities of crude fat and serine as compared with control feed. During broiler finisher phase, the digestibilities of crude protein, fat, fiber, ash and amino acids under study were not affected except for threonine that was significantly higher in 10% PKM as compared with control feed but similar with 5% PKM feed. In growth performance trial, levels of PKM (5% or 10%) did not significantly (P>0.05) affect average body weight, feed consumption and FCR of broiler chickens (Fig. 2). In conclusion, data indicated that PKM inclusion in broiler feed replaced corn and SBM up to 10% without any statistically significant effect on production performance of broiler chickens; however, it increased the feed cost up to 3%.

REFERENCES Abdollahi M. R., Hosking B. J., Ning D., Ravindran V., (2016). Influence of Palm Kernel Meal Inclusion

and Exogenous Enzyme Supplementation on Growth Performance, Energy Utilization, and Nutrient Digestibility in Young Broilers. Asian Australasian. J. Anim. Sci. 29 (4):539-548

Alshelmani M.I., Loh T.C., Foo H.L., Sazili A.Q., Lau W.H., (2016). Effect of feeding different levels of palm kernel cake fermented by Paenibacillus polymyxa ATCC 842 on nutrient digestibility, intestinal morphology, and gut microflora in broiler chickens. Animal Feed Science and Technology. 216:216-224

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ANTIMICROBIAL USE AND RESISTANCE IN COMMERCIAL VILLAGE CHICKENS: A STUDY ON KNOWLEDGE AND PRACTICES AMONG FARMERS

M.I. NUR1, A.K. ARIFAH1*, N.M.A.N.M. FAIZ2 AND M. ROZAIHAN3 1Department of Veterinary Preclinical Sciences, 2Department of Clinical Studies,

3Department of Farm and Exotic Animal Medicine and Surgery, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor


ABSTRACT

The growth of commercial village chicken farming in Malaysia is improving with the involvement of new farmers due to its huge potential. A study was carried out in selected states of Malaysia to determine the practice of antimicrobial usage, the level of awareness on antimicrobial resistance (AMR) among the commercial village chicken farmers, and to identify the factors that are associated with the level of awareness on AMR among farmers. Questionnaire was distributed to the farmers via two methods, farm visit and online survey. The results from this study showed that majority (77%) of the commercial village chicken farmers use antimicrobial in their farms. Most of the farmers have an average level of awareness on AMR (56%). The results also suggested that there is association between the education level with the level of awareness on AMR (P<0.05). Therefore, the level of awareness among farmers was only moderate which means the farmers were not fully aware of AMR. Keywords: antimicrobial resistance, awareness, commercial village chicken.

INTRODUCTION Nowadays, village chicken farming is popular in Malaysia. Before, village chickens were reared as backyard farming and only for self-consumption (Apun et al., 2008) but recently, village chicken has been commercialized as the farmers rear chickens in a large scale as their source of living. Village chicken farming has grabbed the youngsters’ attention as there is active emergence of new young farmers that realized on village chicken potential in Malaysia. Village chicken was claimed or believed to be free of antibiotics but one study done on free-range village chicken in Sarawak in 2008 stated that Escherichia coli (E. coli) isolated from the village chicken was 56% resistant to tetracycline (Apun et al., 2008) and another study in 2016 stated that E. coli isolated from chicken in various part of Malaysia showed that 81.6% multidrug resistance (Roseliza et al., 2016). Based on those findings, there is a possibility that antimicrobials have been used in village chicken farming and antimicrobial resistance can happen due to misuse or overuse of antimicrobial (Saleha, 2007). Therefore, the aims of this study were to determine the level of (i) knowledge on antimicrobial, and practice of antimicrobial usage among farmers, and (ii) awareness on AMR among farmers. MATERIALS AND METHODS A cross-sectional study was conducted using questionnaire. The number of respondents was 25 commercial village chicken farmers from 25 different farms in the selected states of Malaysia: Selangor, Melaka, Johor, Pahang, Kedah and Perak. This study was conducted by distributing the questionnaire to the farmers via two methods, farm visit and online survey. The data collected was tabulated in the Microsoft Excel 2010 and analysed using SPSS statistics version 20 (IBM Corp.). In this study, three tests were used, Multiple Response Sets, Pearson Chi-square (X2) test and Fisher’s Exact Test.


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RESULTS AND DISCUSSION The knowledge of majority of commercial village chicken farmers (56%) on antimicrobial was at an average level as most of them seem to be aware of the term antimicrobial, but they seem not to fully understand or know the meaning of antimicrobials. Most of the commercial village chicken farmers (64%) have their own antimicrobial stock. Ong et al. (2014) reported that E. coli isolated from avian and porcine samples from 2010 to 2013 showed resistance to amoxicillin (92.9%, n=130/140) and florfenicol (54.1%, n=20/37). Majority of the commercial village chicken farmers (60%) in Malaysia had moderate level of awareness on AMR. This finding contradicted with the result of the survey obtained by Etienne et al. (2016) which stated that pig and poultry farmers in European Union (EU) had high levels of awareness on antibiotic resistance as they were aware that a variety of circumstances in the use of antibiotics could lead to antimicrobial resistance. There was a significant association between farmers’ knowledge on antimicrobial and level of awareness on AMR. There was a significant association between education level and farmers’ knowledge on antimicrobial (P<0.05). There was also a significant association between education level and level of awareness on AMR among commercial village chicken farmers. This study finding was similar to Eltayb et al. (2012) that there was a significant relation between the level of education and awareness on antibiotic resistance as the P-value (0.004) was less than α-value (0.05).

REFERENCES Apun K., Chong Y.L., Abdullah M.T. & Micky V. (2008), Antimicrobial susceptibilities of Escherichia coli

isolates from food animals and wildlife animals in Sarawak, East Malaysia, Asian Journal of Animal and Veterinary Advances 3 (6): 409-416.

Eltayb A., Marrone G., Lundborg C. S., Barakat S. & Shaddad S. A. I. (2012), Antibiotic use and resistance in animal farming: a quantitative and qualitative study on knowledge and practices among farmers in Khartoum, Sudan, Zoonoses and Public Health 59(5): 330-8.

Etienne J., Chirico S., Gunabalasingham T., Dautzenberg S. & Gysen S.(2016), EU Insights – Perceptions on the human health impact of antimicrobial resistance (AMR) and antibiotics use in animals across the EU, External Scientific Report, EFSA Supporting Publications, Volume 14, Issue 3: pp.62.

Ong L. P., Muniandy K., How S. P., Tang S. T. P. & Lim B. K. (2014), A report on antibiotic resistance of Eschericia coli isolated from veterinary samples in Malaysia from 2010 to 2013, Malaysian Journal of Veterinary Research, 26th Veterinary Association Malaysia Congress, Volume 5 Supplement: pp 0-71.

Roseliza R., Khairani-Bejo S., Zunita Z., Ramlan M., Khoo E. & Rosnah Y. (2016), Antibiotic resistance of Escherichia coli isolated from chicken in Malaysia, Malaysian Journal of Veterinary Research, Volume 7 No. 1 January 2016: pp 65-76.

Saleha, A. A. (2007), Pathogens and residues: how safe is our meat?, Universiti Putra Malaysia, Universiti Putra Malaysia Press: 1-34.

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FACTORS AFFECTING DRINKING WATER VACCINATION INTAKE IN COMMERCIAL POULTRY FARMS

M.Y.V. CHONG1, N.M. FAIZ1* AND J.E. LEE2

1Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor

2Ceva Animal Health Malaysia Sdn. Bhd. *Corresponding author: [email protected]

ABSTRACT

Vaccines are important means of prevention and control of poultry diseases worldwide. Drinking water vaccination remains as one of the most common mass vaccine administration methods. Its efficiency relies on the vaccinated water consumed by the chickens. This study aims to determine the success rate of drinking water vaccination of commercial poultry farms in Malaysia and to evaluate factors affecting such practice. 5 poultry farms that would undergo drinking water vaccination were selected for study. Prior to vaccination, blue dyes (Cevamune®) were added in the drinking water tank and farm parameters were recorded. 2 hours post vaccination, 100 to 250 chickens (1% of house population) were randomly sampled and evaluated for vaccine intake. From the study, only 10% of the houses fulfilled the requirement of yielding a minimum of 90% positive vaccine intake. Result further revealed that ambient temperature, water restriction hours, stocking density, age of chick, and effort of workers to chase chicks during vaccination, have significantly (p>0.01) affected the outcome of drinking water vaccination which could be due to multiple factors. Existing practice of drinking water vaccine delivery should also be revised to better the outcome of this vaccine administration method. Keywords: Commercial poultry farms, drinking water, vaccination, temperature, stocking density.

INTRODUCTION Different poultry production systems, ranging from backyard to intensive system, have been established to meet the increasing demand for animal proteins (Marangon and Busani, 2006). Apart from feed, the cost of production lies the highest in vaccination. This is because vaccination is important in prevention and control of diseases worldwide (Bell and Willia, 2002). Vaccines can be delivered via various methods of administration. Drinking water vaccination remains as one of the easiest, quickest and most common method of mass vaccine administrations. It is not laborious and requires minimum handling of chicks (Bernardi and Johnson, 2011). However, cases of uneven coverage of flock following drinking water vaccination have been reported and have consequently led to outbreaks of diseases, especially for Infectious Laryngotracheitis (ILT) outbreaks in the USA and worldwide. (Menendez et al., 2014). It is important to note that drinking water vaccination efficiency relies on the vaccinated water consumption by the chicks. To preserve the efficacy of this preferred method of mass administration, the need to research on factors affecting water vaccination intake is warranted to reduce drinking water vaccine failures. MATERIALS AND METHODS Five poultry farms that would undergo drinking water vaccination had been selected for study. Two houses (first and last house) were evaluated in the study. Prior to vaccination, Cevamune® (Ceva Santé Animale, France), that act as blue tracer and as chlorine neutralizing agent, were added in the drinking water tank.

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Farm parameters were recorded in an evaluation form, which included ambient temperature, water restriction hours, stocking density, water volume used for vaccination, and age of chicks. Next, chickens were vaccinated according to the farm routine vaccination practice. 2 hours post vaccination, 100 to 250 chickens (1% of house population) were randomly sampled and evaluated based on scorings for vaccine intake based on the blue dye stain in their mouth. Findings were recorded in a scoring chart. All data were further tested for normality using IBM SPSS Statistics version 20 (IBM Corporation, USA) and analyzed using Kruskal-Wallis Test and Chi-square Test. Significant data were further elucidated using Pairwise Comparison Test. RESULTS AND DISCUSSION A successful drinking water vaccination practice is defined by 90% of birds in flock having well-stained crops and tongues, indication of positive vaccine intake (Fernandez, 2008). In this study, among the ten poultry houses (n=10) selected, only one house (1/10) achieved this rate of success, at 97.11%. Nine out of ten houses (9/10) scored below 90%, and the lowest rate was at 39.80%. Farm parameters recorded were evaluated and it was found that as ambient temperature increased, percentage of vaccinated chicks increased significantly, especially at 34.40°C to 34.87°C. Similar trend was observed in parameters like water restriction duration and stocking density, where evident increase was observed at 250 to 280 minutes and 0.45 to 0.70ft2 respectively. This is because ambient temperature is one of the most important factors affecting water consumption (Vermeulen et al., 2002) and an increase in water restriction hours will result in a higher degree of thirst. In addition, as stocking density increased, space available per chick decreased, leading to a reduction of airflow at the level of flock, reducing the ability of chicks to dissipate body heat (Feddes et al., 2002). Thus, water intake would increase as compensation. This study also shows that younger chicks will result in higher percentage of vaccinated chicks. This is because flock with younger chicks would be smaller in size, occupying less space, which lead to an increase in number of chicks available per drinker. An additional finding also indicated that workers showing effort to chase chicks during vaccination would result in an increase in vaccinated chicks. This action encourages chicks to move towards waterer and allows the detection and immediate correction of clogged waterers (CEVA Santé Animale, 2007). Trends observed from the findings may yield high percentage of vaccinated chicks, but the parameters can be a source of stressor for chickens. Further studies are indicated and this study calls for the need to improve in drinking water vaccine delivery in Malaysia by evaluating existing farm routine practice, to ensure high efficiency of vaccination programme in the future.

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REFERENCES Bell, D.D. and William D.W.J. (2002). Commercial Chicken Meat and Egg Production 5th Edition. Bernardi, E. and Johnson, J. (2011). Drinking Water Vaccination: Easy and Effective. Retrieved from

http://www.pacificvet.co.nz/media/11933/Tech-Bulletin-3-Water-Vaccination.pdf CEVA Santé Animale, A.B.U. (2007). Vaccines and Vaccination in Poultry Production. Ceva Santé

Animale 2007. Feddes, J.J.R., Emmanuel, E.J. and Zuidhoft, M.J. (2002). Broiler performance, body weight variance,

feed and water intake, and carcass quality at different stocking densities. Poultry Science, 81:774–779.

Fernandez, A. (2008). Drinking Water Vaccination - a few simple rules. Ross Tech Notes. Marangon, S. and Busani, L. (2006). The use of vaccination in poultry production. Scientific and

Technical Review of the Office International des Epizooties (Paris), 26(1):265–274. Menendez, K.R., García, M., Spatz, S. and Tablante, N.L. (2014). Molecular epidemiology of infectious

laryngotracheitis: a review. Avian Pathology, 43(2):108–117. Vermeulen, B., De Backer, P. and Remon, J.P. (2002). Drug administration to poultry. Advanced Drug

Delivery Reviews, 54(6):795–803.

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PATHOGENICITY AND IMMUNOGENICITY OF FOWL ADENOVIRUS OF MALAYSIAN ISOLATE ATTENUATED IN CHICKEN EMBRYO LIVER CELLS IN COMMERCIAL BROILER

CHICKENS

M.S. NORFITRIAH1, M. HAIR-BEJO1,2,*, A.R. OMAR1,2, I. AINI1,2 AND M.I. NURULFIZA2,3

1Faculty of Veterinary Medicine, 2Institute of Bioscience, 3Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor.


ABSTRACT

The objective of this study was to determine pathogenicity and immunogenicity of Fowl adenovirus (FAdV) isolate of Malaysia (UPM1137) attenuated in chicken embryo liver (CEL) cells at passage 35 (CELP35) in commercial broiler chickens. The study demonstrated that the FAdV CELP35 isolate caused cytopathic effect (CPE) in the CEL cells with refractile, rounding and clumping of cells within 48 to 72 hours post-inoculation (pi). Four nucleotide (nt) bases changes were detected with 3 amino acids (aa) substitutions in L1 loop of hexon gene. In fiber gene, 4nt bases were substituted in knob region resulting 2aa changes. The CELP35 isolate belongs to FAdV group E serotype 8b and is safe and non-pathogenic in broiler chickens. High FAdV antibody response was detected at day 21pi as a result from the virus-induced immunity which is prominent via parenteral route. It was concluded that the FAdV isolate was attenuated, non-pathogenic and able to induce FAdV antibody response in chickens. Keywords: Fowl adenovirus (FAdV); chicken embryo liver (CEL) cells; pathogenicity; immunogenicity, attenuated. INTRODUCTION Fowl adenovirus (FAdV) serotype 8b has been identified as causative agent of Inclusion body hepatitis (IBH) in commercial broiler and layer chickens in Malaysia (Juliana et al., 2014; Norfitriah et al., 2018). Since FAdV is pathogenic in chickens, it is important to develop vaccine against the virus infection for control and prevention of disease outbreaks. The development of live attenuated FAdV vaccine has been major focus of attempt recently. Cell cultures and chicken embryos routinely used as medium for attenuation of FAdV (Schonewille et al., 2010; Mansoor et al., 2011). In addition, live attenuated FAdV vaccine elicits strong immune response with high protection rate against IBH than inactivated vaccine in chickens (Mansoor et al., 2011). Molecular characterisation and pathogenicity of the virus in chicken are crucial to distinguish between wild and attenuated FAdV isolate. Although hexon and fiber genes encoded for antigenic determinant, yet, the role for virus virulence remained obscure in both proteins. The objective of this study was to determine pathogenicity and immunogenicity of FAdV isolate of Malaysia (UPM1137) attenuated in chicken embryo liver (CEL) cells at passage 35 (CELP35) in commercial broiler chickens. MATERIALS AND METHODS Fowl adenovirus isolate of Malaysia (UPM1137) was originated from an outbreak of IBH and gizzard erosion in commercial layer chickens with 2% mortality. Liver from affected chicken was processed for virus isolation into SPF chicken embryonated eggs for two passages (E2). The isolate (E2) was further adapted and attenuated in chicken embryo liver (CEL) cells for 35th passages (CELP35). Viral supernatant from


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original (E2) and CELP35 was subjected for molecular characterisation based on hexon and fiber genes sequence analysis. The pathogenicity and immunogenicity of CELP35 was determined in commercial broiler chickens. Sixty-four, day-old commercial broiler chicks were divided into three major groups namely; groups A, B and C. Each group was divided further into sacrificed and mortality groups. All chickens in groups A and B were inoculated with 0.5mL propagated FAdV isolate (CELP35) with virus titer of 106.7TCID50/mL via oral and intraperitoneal (IP) route, respectively at day old. The chickens in group C remain uninoculated and acted as the control group. Feed and water were given ad libitum, and the chickens were monitored daily for any abnormal signs until 3-week post-inoculation (pi). Four chickens were sacrificed at day 0pi in group C followed by days 3, 7, 14 and 21pi in all groups. The body weight and blood samples were collected for FAdV antibody detection by ELISA. On necropsy, samples of trachea, liver and gizzard were collected for histological examination. RESULTS AND DISCUSSION The FAdV CELP35 isolate caused cytopathic effect in CEL cells with refractile, rounding and clumping of cells. Substitution of 4 nucleotide (nt) bases was detected in L1 loop of hexon gene with 3 amino acids (aa) changes at position 44, 133 and 185 in CELP35. In fiber gene, 4 out of 6nt bases were substituted in knob region resulting 4aa changes at position 189, 274, 348 and 360. The isolate belongs to FAdV group E serotype 8b. The study demonstrated that the CELP35 was safe and non-pathogenic in commercial broiler chickens. Neither clinical signs nor gross and histological lesions of the trachea, liver and gizzard were observed in all groups throughout the trial. The body weight of chickens in groups A, B and C were significantly increased (p<0.05) from day 0pi to 21pi. Interestingly, the CELP35 isolate induced high antibody response in group B (IP route) at day 21pi (2348 ± 1800) compared to control (69 ± 34) and group A (190 ± 136). The parenteral route stimulates high antibody titer as a result of virus induced immunity without interference by maternal derived antibody (Ojkic and Nagy, 2003). The antigen rapidly absorbed into systemic circulation and multiplied directly in the liver prior induction of antibody response. It was concluded that the propagated FAdV isolate in CEL cells (CELP35) was attenuated, non-pathogenic and able to induce FAdV antibody response in chickens. REFERENCES Ojkić, D. and Nagy, É. (2003). Antibody response and virus tissue distribution in chickens inoculated

with wild-type and recombinant Fowl adenoviruses. Vaccine, 22(1): 42-48. Juliana, M.A., Nurulfiza, M.I., Hair-Bejo, M., Omar, A.R. and Aini, I. (2014). Molecular characterization

of Fowl adenoviruses isolated from Inclusion body hepatitis outbreaks in commercial broiler chickens in Malaysia. Pertanika Journal of Tropical Agriculture Science, 37(4):483-497.

Mansoor, M.K., Hussain, I., Arshad, M. and Muhammad, G. (2011). Preparation and evaluation of chicken embryo-adapted fowl adenovirus serotype 4 vaccine in broiler chickens. Tropical Animal Health and Production, 43: 331-338.

Norfitriah, M.S., Hair-Bejo, M., Omar, A.R., Aini, I. and Nurulfiza M.I. (2018). Molecular detection and pathogenicity of fowl adenovirus isolated from disease outbreak in commercial layer farm. International Journal of Agricultural Sciences and Veterinary Medicine, Vol. 6(1): 73-84.

Schonewille, E., Jaspers, R., Paul, G. and Hess, M. (2010). Specific pathogen free chickens vaccinated with a live FAdV-4 vaccine are fully protected against a severe challenge even in the absence of neutralizing antibodies. Avian Diseases, 54(2): 905-910.

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CHARACTERIZATION OF THREE FOWL ADENOVIRUS ISOLATES OF MALAYSIA PROPAGATED IN CHICKEN EMBRYO LIVER CELLS

C.C. UGWU1,4, M. HAIR-BEJO1,2,*, M.I. NURULFIZA1,3, A.R. OMAR1,2 AND I. AINI1,2

1Faculty of Veterinary Medicine, 2Institute of Bioscience, 3Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 4Department of Animal Science

and Technology, Federal University of Technology, Owerri *Corresponding author: [email protected]

ABSTRACT

The objectives of this study were to characterize fowl adenovirus (FAdV) isolates of Malaysia propagated in chicken embryo liver (CEL) cells. Three isolates of FAdV namely; UPM11142, UPM11134 and UPM08136 were used. They were first propagated in specific pathogen free (SPF) chicken embryonated eggs (CEE) once (E1) and further propagated in CEL cells for 5 consecutive passages (CELP1 to CELP5). The E1, CELP1 and CELP5 of all of the isolates were characterized by PCR technique. The PCR products were then sequenced and analyzed. The study showed that all the isolates caused mortality in SPF CEE eggs and adapted well in CEL cells. Eosinophilic intranuclear inclusion bodies were observed in the cells. The UPM11142 and UPM11134 exhibited more pathogenic in CEE and CEL cells than UPM08136. All of the FADV isolates were characterized as FAdV group E serotype 8b. Nucleotide and amino acid changes were recorded in CELP1 and CELP5 when compared to E1. These isolates have potential to be used for vaccine development against the disease. Keywords: Fowl adenovirus; propagation; characterisation, chicken embryo liver (CEL) cells.

INTRODUCTION Fowl adenovirus (FAdV) causes hydropericadium-hepatitis syndrome with resultant heavy losses in meat type poultry industry worldwide. FAdV is an emerging pathogen in Malaysian poultry industry. It has been reported to cause inclusion bodies hepatitis among broiler flocks in Perak (Hair-Bejo, 2005), and Johore and Malacca (Norina et al., 2016) with 9.6% and 30% mortality, respectively. It causes heavy economic losses in poultry industry worldwide (Ojkic et al., 2008; Dar et al., 2012). One of the challenges of this disease in Malaysia is the unavailability of vaccine for the prevention and control of the disease. Although vaccines have been developed in some countries with varying successes, the need for a Malaysian vaccine developed with similar serotype of the local field virus is pertinent. The objectives of this study were to propagate 3 FAdV isolates of Malaysia in chicken embryo liver (CEL) cells and determine their characteristics for development of vaccine. MATERIALS AND METHODS Three FAdV isolates of Malaysia identified as UPM11142, UPM11134 and UPM08136 were used for this study. Each isolate was first inoculated into specific pathogen free (SPF) chicken embryonated eggs (CEE) via chorioallontoic membrane (CAM). The eggs were incubated at 37oC and observed daily for mortality. Liver and CAM of dead embryo were harvested and homogenized, and the homogenate was centrifuged at 1500rpm for 5 mins (Embryo passage 1 (EP1)). The supernatant was used to infect chicken embryo liver (CEL) cells. The CEL cells were prepared from 13-15 days old SPF eggs in 25m2 tissue culture flasks. The EP1 virus inoculum (0.1ml) was inoculated onto 80% confluent layer of CEL cells, incubated and observed daily for cytopathic effects (CPE). At 100% CPE, the cells were harvested, freeze-thawed thrice at -20oC


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and centrifuged at 1500rpm for 5 mins. The supernatant (CELP1) was then used to infect the next CEL cells and the process repeated till passage 5 (CELP5). The CELP1 and CELP5 of each isolate were examined for inclusion bodies by HE staining. The DNA from EP1, CELP1 and CELP5 of each isolate were extracted and used for PCR targeting hexon and fibre genes. The PCR products were purified, sequenced and analysed. RESULTS AND DISCUSSION The UPM11142 and UPM11134 FAdV isolates caused 100% mortality to SPF CEE within 6-9 days post inoculation (pi) while UPM08136 caused 85.7% mortality at day 11 pi. The dead embryo showed typical FAdV lesions which includes friable, discoloured and inflamed liver with haemorrhages. Typical CPE of FAdV on CEL cells were surface shining, rounding, clumping, enlargement and detachment of CEL cells. The UPM08136 and UPM11134 isolates (CELP1) caused CPE at days 4 and 5 pi, respectively. However, UPM11142 isolate did not produce CPE at CELP1. At CELP5, CPE were observed within 24 hours in UPM11142 infected flasks, and at days 2 and 3 for UPM11134 and UPM08136, respectively. Histological examination showed intra-nuclear inclusion bodies (INIB) in the CEL cells (Soumyalekshmi et al., 2014). The study showed that FAdV UPM11142 and UPM11134 exhibited more pathogenic in CEE and CEL cells than UPM08136. All the 3 isolates were positive for hexon and fibre genes and were confirmed as FAdV serotype 8b (Meulemans et al., 2001). Various nucleotide and amino acid changes were observed in CELP1 and CELP5 isolates when compared to EP1. They were 98-100% related to UPM04217, a FAdV isolate of Malaysia. It suggests that the FAdV outbreaks in Malaysia are probably from the same origin resulting to infection from one chicken farm to the other farms. In conclusion, the 3 FAdV isolates of Malaysia were able to adapt well in CEE and CEL cells. The propagation of the FAdV in CEL cells could lead to attenuation and the development of vaccine against the disease. REFERENCES Dar A., Gomis S., Shirley I., Mutwiri G., Brownlie R., Potter A., Gerdts V. and Tikoo S.K. (2012).

Pathotypic and molecular characterization of a Fowl adenovirus associated with Inclusion body hepatitis in Saskatchewan chickens. Avian Diseases, 56:73-81.

Hair-Bejo M. (2005). Inclusion body hepatitis in a flock of commercial broiler chickens. Journal of Veterinary Malaysia, 17:23-26.

Meulemans G., M. Boschmans, T.P. van den Berg & M. Decaesstecker (2001). Polymerase chain reaction combined with restriction enzyme analysis for detection and differentiation of fowl adenoviruses, 30(6): 655-660.

Norina L., Norsharina A., Nurnadiah A.H., Redzuan I., Ardy A. and Nor-Ismaliza I. 2016. Avian adenovirus isolated from broiler affected with inclusion body hepatitis. Malaysian Journal of Veterinary Research, 7(2):121-126.

Ojkic D, Martin E, Swinton J, Vaillancourt JP, Boulianne M and Gomis S. (2008). Genotyping of Canadian isolates of fowl adenoviruses. Avian Pathol 37: 95–100.

Soumyalekshmi S., M. K. Ajith and Meshram Chandraprakash (2014). Isolation of fowl adenovirus in chicken embryo liver cell culture and its detection by hexon gene based PCR. Ind. J. Sci. Res. and Tech. 2(3):33-36.

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THE STUDY OF MEAT QUALITY IN MALAYSIAN TURKEY

N.J.S. HAWA AND I.H. LOKMAN1,* 1Department of Veterinary Preclinical Sciences, Faculty of Veterinary Medicine,

Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia *Corresponding author: [email protected]

ABSTRACT

This study was conducted to investigate the meat quality of Malaysian turkey (Meleagris gallopavo) that have received little attention from the researchers. Malaysian turkey or known as well as ‘Ayam piru’ carcass was subjected to an experiment on meat quality in which involved of carcass composition, pH, color, cooking loss, meat tenderness and proximate analysis. The carcass composition experiment was conducted according to dissection method by using the whole carcass while breast muscle was used for pH, color, cooking loss, muscle tenderness and proximate analysis which include of ash, dry matter, crude protein, crude fibre and fat. At the age of 50 days, 5 carcasses of Malaysian turkey were obtained. Percentage for carcass composition showed that 12.5% was skin and fat, 18.2% was bone and 35.2% was muscle. The pH for meat turkey was 5.6 which slightly acidic. Cooking loss for meat was 10% and colour result showed that L* 57.32 a* 9.09 b* 12.89. Proximate analysis showed that ash content 1.6%, dry matter and moisture 97.2% and 2.7%, crude fat 11.6%, crude fibre 10% and crude protein was 23%. Keywords: Carcass composition, colour, cooking loss, Ayam piru, Meleagris gallopavo. INTRODUCTION Poultry are domesticated birds from members of the order Galliformes (chickens, quails and turkeys) and Anseriformes that consist of family Anatidae or commonly known as “water fowl” (Eaton, 1992). Wild turkeys (Meleagris gallopavo) are one of the most widely distributed game bird species in North America and found throughout most of the eastern and western United States. Wild turkeys have been introduced to Germany and New Zealand (Eaton, 1992). Turkey meat is less consumed in Malaysia compared to western countries such as Europe and United States. Turkey grows faster like broiler chickens and become suitable for slaughter purpose within a short period. Commercial turkey farming is a profitable business idea. Most people eat broiler chickens that are available in many places and the price is more reasonable than turkey meat. In Malaysia, turkey or locally known as ‘ayam piru’ is not new in the poultry industry. However, the lack of awareness makes turkey meat unpopular as a delicacy or dish. The study of meat quality in Malaysian turkey had received little attention from the researchers. Thus, the aim of study is to establish the meat quality of turkey meat in Malaysian turkey by measuring the pH, color, tenderness and also carcass composition. Besides, nutritional quality of the turkey meat also being evaluated by using proximate analysis which include of dry matter, ash, crude protein, crude fibre and fat. MATERIALS AND METHODS Five (n=5) female Malaysian turkey (Meleagris gallopavo) have been randomly selected. These turkeys are from AAF agro farm in Simpang Renggam, Johor that applied semi intensive system with a population of almost 90 turkeys. At the age of 50 days, they were randomly taken and slaughtered according to a halal slaughter procedure. Carcass composition measurement and the whole carcass for each turkey


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was weighted and recorded. The procedure was followed by removing the fat, the meat and bone, weighted and recorded. The pH analysis was performed by using portable pH meter that directly inserted into the meat. Colour analysis was analysed on pectoralis muscle by using Chroma meter CR 410 (Konica Minolta, Jpn.; based on CIElab method). Cooking loss was calculated by the loss of samples weighted before (w1) and after cooking (w2). The breast meat was kept in air tight plastic bag and cooked in the water bath at temperature 80°C for 20 min. Tenderness was analysed by using breast meat from the cooking loss procedure performed using Stable Micro System Texture Analyzer (Model TA-XT plud, UK) equipped with a flat Volodkovich shear blade. Proximate analyses were carried out according to certification procedures outlined by the Manual of Laboratory Techniques, UPM. RESULTS AND DISCUSSION The mean for carcass composition for bone, muscle and skin, and fat were from 18.22%, 35.17% and 12.45% respectively. Excessive energy intake leads to increasing fat deposition in the body (Moravej et al., 2006). The pH analysed mean is 5.684, this acidic pH value of the meat is results from the process of rigor mortis (Kim et al., 2016). Mean for cooking loss was 12%, Moutney and Arganosa (1989) reported that increasing of fat percentage will increase the percentage of cooking loss. Tenderness mean is 45.092, Kim et al, (2016) stated that mainly the tenderness is due to the effect of increase glycolytic fiber contents in the muscle of faster growing animals. Dry matter percentage was 97%, young turkeys had higher dry matter in the breast meat than older turkeys. Ash percentage is 1.6 %, young turkeys had higher ash in the breast meat than older turkeys. Crude protein percentage is 23%, value for protein is varied between 24-25%. Crude fat percentage was 11.6% which supported with study from Fernandez et al. (2001) and Werner et al. (2008), decreasing of moisture percentage will increase the value of fat percentage in turkey meat. Crude fibre percentage is 10%, Werner et al. (2008) stated that increasing of moisture percentage will increase the fiber percentage. REFERENCES Eaton, S. (1992). Wild Turkey (Meleagris gallopavo). In: Poole P., Stetthenheim P. and Gill F. (eds). The

Birds of North America, Vol. 22. The Academy of Natural Sciences, Philadelphia. Washington, DC: The American Ornithologists’ Union.

Fernandez, X., Santé, V., Baeza, E., Le Bihan-Duval, E., Berri, C., Rémignon, H., Babilé, R., Le Pottier, G., Millet, N., Berge, P., and Astruc, T. (2001). Postmortem muscle metabolism and meat quality in three genetic types of turkey. British Poultry Science. 42:462-469.

Kim, H., Yan, F., Hu, J., Cheng, H. and Kim, Y. (2016). Effects of probiotics feeding on meat quality of chicken breast during postmortem storage. Poultry Science. 95(6): 1457-1464.

Moravej, H., Khazali, H., Shivazad, M. and Yeganeh, H.M. (2006). Plasma concentrations of thyroid hormone and growth hormone in Lohmann male broilers fed on different dietary energy and protein levels. International Journal of Poultry Science. 5: 457-462.

Mountney, G. J. and Arganosa, F.C. (1989). The effect of phosphates on moisture absorption, retention and cooking losses of broiler carcasses. Poultry Science. 42: 384-88.

Werner, C., Riegel, J. and Wicke, M. (2008). Slaughter performance of four different turkey strains, with special focus on the muscle fiber structure and the meat quality of the breast muscle. Poultry. Science. 87:1849-1859.

https://scialert.net/fulltext/?doi=ajas.2009.13.17#78301_ja

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PATHOGENICITY OF SALMONELLA STANLEY ISOLATE IN COMMERCIAL BROILER CHICKENS

R. BALQIS1 AND M. HAIR-BEJO1,2,*

1Faculty of Veterinary Medicine, 2Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia


ABSTRACT

Salmonella Stanley (S. Stanley) is gram negative rods of the Enterobacteriaceae family commonly found in ducks, a reservoir of the bacteria. The potential risk for exposure to Salmonella via contaminated food rises with growth in consumption of meat and poultry products. The objectives of this study were to determine pathogenicity of S. Stanley isolate (UPM517) in chickens and to isolate the agent from the organs. The study showed that body weight of chickens in inoculated (A) and controlled (B) groups increased throughout the trial with no significant difference (p>0.05) between groups. No abnormal clinical signs were recorded in all groups, except on day 6 post inoculation (pi) one chick (1/8 or 12.5%) from mortality group A showed weakness and was found dead on the next day (day 7 pi). Necropsy revealed no significant findings in all groups, except splenomegaly in chicks from group A on day 4, 7 and 14 pi. Histologically the liver remained normal in all group of chickens. Salmonella was isolated from group A on day 1 pi from liver and caecal tonsils; on day 4 pi from liver, caecal tonsils and spleen; on day 7 pi from liver, spleen, caecal tonsils, cloacal swab; but on day 14 pi Salmonella was not isolated. No Salmonella was isolated from group B. Salmonella antibody titre declined in all groups throughout the trials with no significant difference (p>0.05) in the titre between groups. In conclusion, the S. Stanley isolate is pathogenic in commercial broiler chickens. Keywords: Salmonella Stanley, commercial broiler chickens, pathogenicity.

INTRODUCTION Salmonella Stanley (S. Stanley), also known as S. enterica subspecies enterica serovar Stanley, is gram negative rods of the Enterobacteriaceae family that commonly cause human foodborne disease, with ducks is the common reservoir for the bacteria (Bangtrakulnonth et al., 2004). Salmonella Stanley infections were among the 15 most common serovars in 12 out of 104 countries (Bangtrakulnonth et al., 2004). A study in Thailand showed from 2002 to 2007, the second most common serovar causing human salmonellosis cases is due to S. Stanley and also prevalent for human Salmonellosis in a few other countries (Hendriksen et al., 2012). However, not much is known regarding S. Stanley infection in chickens. The objectives of the study were to determine the clinical signs, mortality percentage, gross and histological lesions, and Salmonella antibody titre of chickens inoculated with S. Stanley isolate. MATERIALS AND METHODS A total of 52 days old chicks was divided into two groups; groups A (24 chicks) and B (28 chicks). The chicks in group A were inoculated intraperitoneally with 0.1 ml of 1x10⁸ colony forming unit (cfu) of S. Stanley at day old, whilst the group B which were remain uninoculated (control group). The S. Stanley isolate (UPM517) was obtained from 21 days old healthy broiler chickens in Perak, Malaysia. The chicks in each group were further divided into two groups by separating 8 chicks from each group for mortality group and the rest of the chicks were placed in sacrifice group. The chicks were provided with feed and water ad libitum and monitored for abnormal clinical signs and mortality at least twice daily throughout the trial. At the


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beginning of experiment prior to bacterial inoculation, four chicks from the control group were sacrificed. Four chicks from each group A and B were sacrificed on day 1, 4, 7 and 14 post inoculation (pi). Prior to necropsy body weights were recorded and blood samples were collected for detection of Salmonella antibody using ELISA technique. On necropsy, gross lesions were recorded and samples of liver were fixed in 10% buffered formalin for histological examination. Samples of liver, spleen, caecal tonsils and cloacal swabs were collected for bacterial isolation and identification. RESULTS AND DISCUSSION The study revealed that the body weight of chickens was increased from 51.8±0.75g on day 0 pi to 512.5±7.7g on day 14 pi in the control group. A similar pattern of increment was observed in the inoculated group. No abnormal clinical signs and gross lesions were recorded in all chickens in both groups, except splenomegaly were seen from chicks in group A at day 4, 7 and 14 pi. One chick (1/8 or 12.5%) from the mortality group A showed weakness on day 6 pi and died on the next day. Intracellular localization Salmonella bacteria that survived despite being phagocytosed by macrophages and cellular proliferation in organ occurred as an instant response to invasion of pathogen resulting in enlarged organs such as splenomegaly (Al-Abadi and Al-Mayah, 2013). However, histology revealed no lesion in liver in all groups. Salmonella was isolated from liver and caecal tonsils at day 1, 4 and 7 pi, from spleen at days 4 and 7 pi and from cloacal swab at day 7 pi. It was recovered from liver, spleen and caecal tonsils of a dead chicken at day 7 pi. Liver and spleen were the indication of systemic infection; caecal tonsils showed intestinal colonization; and cloacal swab was sign of shedding (Gast, 2013). The cause of death in the chicken is likely due to septicaemia (Henderson et al., 1999) and burden with organism in the organs that cause multiple organ failure. Salmonella antibody titre decreased from 401±208 on day 0 pi to 7±2 on day 14 pi for the inoculated group A. A similar pattern of antibody titre (p>0.5) was recorded in the control group. The decrement of antibody titre can be explained by maternal antibody received by the chicks from the immunized hens as a result of vaccination or previous exposure of salmonellosis (Gast, 2013). Therefore, presence of maternal antibody might interfere with the ongoing infection. In conclusion, S. Stanley isolate is pathogenic in broiler chickens following inoculation at day old of age. REFERENCES Al-Abadi, I.K.M. and Al-Mayah, A.A.S. (2013). Observations on the pathogenesis of orally inoculated

Salmonella typhimurium in a newly hatched broiler chicks. Iraqi Poultry Sciences Journal. 7(1):42-51.

Antunes, P., Mourão, J., Campos, J. and Peixe, L. (2016). Salmonellosis: the role of poultry meat. Clinical Microbiology and Infection. 22(2): 110-121.

Bangtrakulnonth, A., Pornreongwong, S., Pulsrikarn, C., Sawanpanyalert, P., Hendriksen, R.S., Lo Fo Wong, D.M.A. and Aarestrup, F.M. (2004). Salmonella serovars from humans and other sources in Thailand, 1993-2002. Emerging Infectious Diseases. 10(1):131-136.

Gast, R.K. (2013). Salmonella Infections. In: D.E. Swayne, J.R. Glisson, L.R. McDougald, L.K. Nolan, D.L. Suarez, and V. Nair (Eds.), Diseases of Poultry (13th ed., pp. 693–706). Oxford, UK: John Wiley and Sons, Inc.

Henderson, S.C., Bounous, D.I. and Lee, M.D. (1999). Early events in the pathogenesis of avian Salmonellosis. Infection and Immunity. 67(7): 3580-3586.

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SEQUENCE ANALYSIS OF S1 GENE OF MALAYSIAN INFECTIOUS BRONCHITIS VIRUS VARIANT AND QX-LIKE STRAINS FOLLOWING SERIAL PASSAGE IN SPECIFIC-

PATHOGEN-FREE EMBRYONATED CHICKEN EGGS

M.I. ISMAIL1, M. SUWAIBAH1, S.W. TAN1, M. HAIR-BEJO1,2 AND A.R. OMAR1,2,* 1Institute of Bioscience, 2Faculty of Veterinary Medicine, Universiti Putra Malaysia,

43400 UPM Serdang, Selangor, Malaysia *Corresponding author: [email protected]

ABSTRACT Infectious bronchitis virus (IBV) causes infectious bronchitis (IB) disease in chickens is primarily a disease associated with respiratory tract. However, depending on virus strains, IBV can also infect the gut, reproductive and kidney of chickens. Controlling the disease is problematic because of the presence of diverse serotypes and strains, and poor cross-protection against different serotypes/strains. Recently, we have characterized the IBV strains circulating in Malaysia. Two of the prevalent strains, Malaysian local variant (MLV) (IBS037A/2014) and QX-like (IBS130/2015) were attenuated by serial passage of the virus in SPF embryonated chicken eggs. The virus strains at different passages were characterized based on sequence analysis of the partial S1 gene. Overall, IBS130/2015 showed a higher number of nucleotide and amino acid variations compared to IBS037A/2014 following serial passage in embryonated chicken eggs. Keywords: Infectious bronchitis, live attenuated, molecular characterization.

INTRODUCTION Avian coronavirus infectious bronchitis virus (IBV) is the causative agent of infectious bronchitis (IB) disease in chickens, a highly contagious disease of the upper respiratory tract in chickens. However, some IBV strains can cause nephritis which associated with high mortality. Controlling the disease is challenging due to the presence of different IBV strains that show partial cross-protection against each other, and only some strains show the evidence of protectotypic (Cook et al., 2012). Hence, effective control of IB requires the use of vaccine strains that able to provide protection against field IBV (Cavanagh and Gelb, 2008). Recently, we have identified two predominate nephropathogenic IBV strains, MLV and QX-like circulating in poultry farms in Malaysia (Khanh et al., 2017). A preliminary study indicated the currently available vaccines unable to provide full protection against these two strains. The S1 subunit of S protein is responsible for attachment of IBV to host cell receptor and contains epitopes that can induce neutralization (Cavanagh, 2007). In addition, the differences of 2 to 3% (10 to 15 amino acids) in the S1 region may produce a new IBV serotype (Cavanagh, 2003). The ultimate objective of this study is to develop a live attenuated IBV vaccine from the MLV and QX-like strains. In this study, we reported the sequence analysis of the partial S1 gene of the Malaysian IBV strains following passaging the viruses in SPF embryonated chicken eggs. MATERIALS AND METHODS Two IBV isolates used in this study, MLV (IBS037A/2014) and QX-like (IBS 130/2015) were attenuated by serial passage the virus in SPF embryonated chicken eggs. Allantoic fluids collected from the inoculated eggs were extracted for viral RNA using the RNeasy® Plus Mini Kit (QIAGEN, Germany) according to the manufacturer’s instructions. The extracted RNAs were subjected to cDNA synthesis and RT-PCR

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amplification. The cDNA was generated by reverse transcription using SensiFAST cDNA Synthesis Kit (Bioline, UK) following the procedures as recommended by the manufacturer’s instructions. The cDNA produced was used as a template to amplify a partial S1 gene of 464 bp fragment using the KAPA HiFi HotStart PCR kit (KAPA Biosystems Pty Ltd., South Africa). The final amplified PCR products were electrophoresed in 1% TAE agarose gel and purified using a MEGAquick-spinTM Total Fragment DNA Extraction (iNtRON Biotechnology, South Korea). The PCR products were submitted for gene DNA sequencing using Sanger method (First BASE Laboratories Sdn. Bhd., Malaysia). Nucleotide sequence and deduced amino acid sequences were aligned and edited using ClustalW and MEGA software Version 6.0.6. The identity of nucleotide and amino acid sequences at different passages were compared using the BioEdit Software version 7.1.3.0. RESULTS AND DISCUSSION As shown in Table 1, the Malaysian QX-like strain, IBS130/2015 showed a higher number of nucleotide variations compared to the variant strain, IBS037A/2014 following serial passage in embryonated chicken eggs. Our results show that IBS037A/2014 has 95.6 to 99.7% nucleotide sequence identity with the highest similarity between virus passage 30 and 50 (99.7%) and the lowest similarity between virus passage 6 and 10 (95.6%). On the other hand, the sequence identity of IBS130/2015 range between 75.9 to 98.2% with the highest similarity between virus passage 30 and 50 (98.2%) and the lowest similarity between virus passage 8 and 50 (75.9%). The amino acids identity of IBS037A/2014 and IBS130/2015 ranged between 94.0 to 99.3% and 50.0 to 96.6%, respectively. In IBS037A/2014, nine amino acids substitutions were observed among the different virus passages between the positions 235 to 387. In IBS130/2015, 76 amino acids substitutions were observed between the positions 238 to 390. Sequence analysis of TW I isolate (2296/95 strain) between passage 4 and 76 found six substitutions of S1 amino acid at the position of 47, 56, 63, 117, 131 and 161 (Huang and Wang, 2006). Hence, attenuation of IBV strains associated with multiple amino acid variations in the S1 gene. Characterization of the amino acid substitutions and pathogenicity study of the attenuated isolates will provide valuable insight into the development of live attenuated IBV vaccine. Table 1. Comparison of the percent nucleotide (bottom and left) and amino acids (top and right) identities sequence of the partial S1 protein of IBS037A/2014 and IBS130/2015

1 2 3 4 5 6 7 8

1. IBS037A P6 - 94.0 96.0 96.6 46.7 89.4 90.1 90.7 2. IBS037A P10 95.6 - 96.6 97.3 46.0 89.4 91.4 92.1 3. IBS037A P30 98.0 96.9 - 99.3 45.3 88.8 90.7 90.1 4. IBS037A P50 98.2 97.1 99.7 - 45.3 89.4 90.1 90.7 5. IBS130 P8 77.0 74.7 76.4 76.6 - 50.0 50.6 50.3 6. IBS130 P10 96.0 94.1 96.5 96.7 76.5 - 94.6 96.0 7. IBS130 P30 95.6 95.4 96.5 96.3 76.3 96.4 - 96.6 8. IBS130 P50 95.4 96.3 95.8 96.1 75.9 96.2 98.2 -

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The identity of IBS037A/2014 and IBS130/2015 in the amino acid levels range between 94.0 to 99.3% and 50.0 to 96.6%, respectively. In IBS037A/2014, nine amino acids substitutions were observed among the different virus passages between the positions 235 to 387. In IBS130/2015, 76 amino acids substitutions were observed between the positions 238 to 390. Previous study has shown that serial passage of TW I isolate (2296/95 strain) between passage 4 and 76 show six substitutions of S1 amino acid at the position of 47, 56, 63, 117, 131 and 161 (Huang and Wang, 2006). Characterization of the amino acid substitutions and pathogenicity study of the attenuated isolates will provide valuable insight into the development of live attenuated IBV vaccine ACKNOWLEDGEMENT This study was supported by HICOE grant, Ministry of Higher Education, Malaysia. REFERENCES Cavanagh, D. (2003). Severe acute respiratory syndrome vaccine development, experiences of

vaccination against avian infectious bronchitis coronavirus. Avian Pathology, 32:567-582. Cavanagh, D. (2007). Coronavirus avian infectious bronchitis virus. Veterinary Research, 38: 281-297. Cavanagh, D. and Gelb, Jr.J. (2008). Infectious bronchitis. In: Diseases of Poultry, 12

th ed., Y.M. Saif,

A.M. Fadly, J.R. Glisson, L.R. McDougald, L.K. Nolan and D.E. Swayne (Eds.), pp: 117-135. Iowa, USA: Blackwell Publishing.

Cook, J.K.A., Jackwood, M. and Jones, R.C. (2012). The long view: 40 years of infectious bronchitis research. Avian Pathology, 41: 239-250.

Huang, Y.P. and Wang, C.H. (2006). Development of attenuated vaccines from Taiwanese infectious bronchitis virus strains. Vaccine, 24: 785-791.

Khanh, N.P., Tan, S.W., Yeap, S.K., Satharasinghe, D.A., Hair-Bejo, M., Bich, T.N. and Omar, A.R. (2017). Molecular characterisation of QX-like and variant infectious bronchitis virus strains in Malaysia based on partial genomic sequences comprising the S-3a/3b-E-M-Intergenic region-5a/5b-N gene order. Avian Diseases, 61: 442-452.

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DETECTION OF GASTROINTESTINAL PARASITES IN JAPANESE QUAILS (CORTUNIX CORTUNIX JAPONICA)

L. NURSAKINAH1, M.N.M. HEZMEE1,*, I.H. LOKMAN1 AND N.M.M. ISA2

1Department of Veterinary Preclinical Sciences 2Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400

UPM Serdang, Selangor, Malaysia *Corresponding author: [email protected]

ABSTRACT

Quails are known for its ability to have high resistance toward common poultry diseases. Therefore, the health status and prevalence of diseases affecting quails is often neglected. One of the most common diseases that affect poultry is endoparasites infection. In Malaysia, currently there is no study or published data in detection of gastrointestinal parasites in Japanese quails. Hence, this study was conducted to detect gastrointestinal parasites in Japanese quails. 30 quails were purchased from Farm X, Malaysia. For this purpose, clinical signs of parasitism were observed through physical examination, fecal examination via Modified McMaster technique, morphological identification via compound microscope and gross lesions via necropsy were performed. Physical examination revealed no clinical signs of parasitism. There was presence of Eimeria oocysts in pooled fecal sample from Farm X. Three common Eimeria species in Japanese quails were found and identified as E. tsunodai, E. uzura and E. bateri. Necropsy revealed normal serosa and mucosa of gastrointestinal tract without presence of any parasites. In conclusion, the presence coccidiasis in this study might indicate natural infection for Japanese quails which beneficial to stimulate immune response in body. Keywords: Japanese quails, gastrointestinal parasites, coccidiosis, E. tsunodai, E. uzura, E. bateri.

INTRODUCTION Gastrointestinal parasites such as nematodes, cestodes and sporozoa can infect quails worldwide. For example, nematodes such as Heterakis gallinarum, cestodes such as Raillietina echinobothrida, Raillietina cysticillus, Choanotaenia infundibulum and Hymenolepis, sporozoa such as Coccidia and Crytosporidium. The importance of this study is to know whether Japanese quails in Farm X, Malaysia show susceptibility or resistance towards gastrointestinal parasites infection. If quails show susceptibility, prevention and control program could be initiated such as extensive biosecurity measure and deworming program. If quails not susceptile, farmers will get the financial benefit on the treatment cost of anti-parasite. MATERIALS AND METHODS Physical examination was done as flock examination and individual examination. Clinical signs of parasitism were observed on all 30 quails. Next, Fecal Egg/Oocyst Count via Modified McMaster Technique was performed. 4g of pooled faecal sample from 30 quails were collected and 56ml of saturated NaCl was added. Then, the filtrates were pipette enough to load in both counting chambers of the Mc Master slide by using pipette. Next, Mc Master slide counting chamber was seen under compound microscope to count the eggs or oocysts seen within the grids. Lastly, all observations and result were recorded and eggs/oocysts per gram of faeces was calculated. In addition, oocyst morphological identification was performed by using compound microscope under x40 objectives (Motic 2.0 Image Plus). Parameters such as shape, length, width, shape index, polar granule and oocyst wall were measured


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and recorded (Berto et. al, 2013). Lastly, post mortem was carried out for all 30 quails. Gross lesions on gastrointestinal tract and presence of helminths such as nematodes or cestodes were examined. RESULTS AND DISCUSSION This study showed presence of gastrointestinal parasite infection in Japanese quails due to Eimeria species. There was presence of coccidian occysts in fecal examination. According Ettinger and Feldman (1995), animal that have natural infection of coccidia can show no clinical signs. This is supported by Texeira (2004) who reported that natural infection was featured as subclinical infection which means Japanese quails can be infected with coccidiosis but asymptomatically. The Eimeria sp. oocysts count for House A and House B of total 30 quails were 20, 272 oocysts per g of faeces and 20, 393 oocysts per g of faeces respectively. In quails, currently there is no study about standard values of calculated OPG number to determine the level and severity of coccidiosis infection. Umar (2014) claimed that high number of oocysts in fecal examination does not determine a serious pathogenic condition because difference in host genetics, nutritional factors, concurrent diseases, and species of the Eimeria. In addition, coccidian oocysts found in this study might indicate normal flora which helps inhibit other pathogenic coccidian in gastrointestinal tract. From the morphological of oocyst identification, we suspected all three common species of Eimeria were presence in all 30 Japanese quails such as E. tsunodai, E. uzura, E. bateri. Three most common Eimeria species found in Japanese quails were reported as low to mild pathogenicity. This morphological identification was compared in the guidelines reported by Berto et. al (2013). Other Eimeria sp. oocysts were hardly to identify. This is because Eimeria oocysts that does not fall in the guidelines might be from other species of Eimeria which are commonly found in quails such as E. dispersa, E. cortunicis, E. taldykurganica, E. tahamensis, E. flumenesis. Identification of different Eimeria species based on morphology of oocysts is very challenging and requires expertise (Umar, 2014). From post mortem examination, there was no significant gross lesion and absence of other parasites along the gastrointestinal tract. This is because of the prolong life cycle of parasites that unable to complete its life cycle within the range and cause diseases. This result was significant since all 30 Japanese quails were healthy without clinical signs of parasitism. Also, there might be insufficient numbers of parasites replication in the gastrointestinal tract to cause pathological lesion. It was concluded that, the presence coccidiasis in this study might indicate natural infection for Japanese quails which beneficial to stimulate immune response in body. REFERENCES Berto, B. P., Borba, H. R., Lima, V. M., Flausino, W., Teixeira-Filho, W. L., & Lopes, C. W. G. (2013).

Eimeria spp. from Japanese quails (Coturnix japonica): new characteristic features and diagnostic tools. Pesquisa Veterinaria Brasileira, 33(12), 1441-1447.

Ettinger, S.J and Feldman, E.C. (1995). Textbook of Veterinary Internal Medicine (Fourth edition). W.B. Saunders Company. pp 59-60

Teixeira, M., Teixeira Filho, W. L., & Lopes, C. W. G. (2004). Coccidiosis in Japanese quails (Coturnix japonica): characterization of a naturally occurring infection in a commercial rearing farm. Revista Brasileira de Ciência Avícola, 6(2), 129-134.

Umar, H. A. (2014). Occurrence Morphometric Identification and Histopathological Lesions of Eimeria Species in Japanese Quails (Coturnix coturnix japonica) in Zaria, Nigeria.

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DETECTION OF HAEMOPARASITES IN JAPANESE QUAIL (CORTUNIX CORTUNIX JAPONICA)

A.N.S. SHAREEN1, M.N.M. HEZMEE1,*, I.H. LOKMAN1 AND N.M.M. ISA2

1Department of Veterinary Preclinical Sciences 2Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400

UPM Serdang, Selangor, Malaysia *Corresponding author: [email protected]

ABSTRACT

Quails are known for its small size and unique qualities of low feed requirements, rapid growth, short gestation, making it an alternative for cheap source of protein other than chickens. However, there is limitation regarding the study of disease in quail production in Malaysia. In the present study, it focuses on detection of haemoparasites in Japanese quails (Cortunix coturnix japonica). Total of 30 quails were taken from different housing, consisting of 25 to 35 days of age. A thorough physical examination was done but all quails do not exhibit any typical signs such as emaciation, loss of appetite, listlessness, difficulty in breathing, and weakness. Blood sample collected for thin blood smear stained revealed that, both houses demonstrated the presence of Plasmodium sp. and Leucocytozoon sp. The results were as follows; high detection of Plasmodium sp. was noted;93.3% in house A and 100.0% in house B and low detection of Leucocytozoon sp.; 40.0% in house A and 46.7% in house B. Post-mortem examination shows no significant changes. In conclusion, there is presence of haemoparasites but absent of clinical signs and significant changes in the organs of affected quails. The current report will improve the understanding on effect of haemoparasites among quails. Keywords: Japanese quails, haemoparasites, Plasmodium sp., Leucocytozoon sp.

INTRODUCTION In worldwide, Japanese quails can be found in many countries which influenced by its good performances in meat and egg production described as small size, low feed requirements, short life cycle, good reproductive potential, good meat taste, better laying ability, rapid growth rate, shorter time of hatching and resistant to common poultry diseases as compared with different species of poultry. Thus, making the quail production an important poultry business. However, there are limitations of studies in quail production which includes; occurrence of infectious disease and non-infectious disease. Based on this study, it focuses on infectious disease involving haemoparasites. In haemoparasites, the most common species found in quails is Leucocytozoon sp., Plasmodium sp. and Haemoproteus sp. Hence, this study was designed for detection of haemoparasites infection in Japanese quails (Cortunix cortunix japonica) in determine the presence of haemoparasites in quail and its clinical signs and post-mortem lesions of quail with haemoparasites infection. Additionally, the hypothesis for this study is there is presence of haemoparasites in Japanese quails. MATERIALS AND METHODS Total of 30 Japanese quails were randomly picked from the marketing age of 28 to 35 days of age from Puyumas FarmBest Sdn Bhd farm and isolated into House A and House B. A thorough physical examination was executed to detect any clinical signs associated with haemoparasites infections which includes checking the body such as its vital signs, body condition score, mentation of animals, posture and gait including


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the presence of ectoparasites. Blood sampling were taken from the brachial vein of each quail and kept in ethylene-diamine-tetra acetic acid (EDTA) tubes and plain tubes for the use of thin blood smear analysis and molecular study using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Post-mortem examinations were carried out after slaughtering of all 30 quails via halal slaughtering to check for any significant lesions. Examination initially checks for external parasites and lesion before opening the carcass. Then the examinations of the organs continued at in situ emphasizing on the liver, spleen, kidney and muscle tissue. Laboratory diagnosis continues at demonstration of type of hemoparasites via 30 thin blood smears stained with Giemsa stain. A smear with no parasites present will be considered negative. Demonstration of hemoparasites genes and determination of hemoparasites species continued using RT-PCR method. RESULTS AND DISCUSSION Thin blood smear demonstrated positive presence of hemoparasites. Plasmodium sp. and Leucocytozoon sp. were detected, resulting in 93.3% (14/15) in house A and 100.0% (15/15) in house B for Plasmodium sp. and 40.0% (6/15) in house A and 46.7% (7/15) in house B for Leucocytozoon sp. Even there is positive infections of haemoparasites, physical examination and post-mortem examination shows no significant findings. This might be indicating that quails can compensate even if its positive for infection. RT-PCR was used as a gold standard in detection of hemoparasites by detecting its gene and identify specific species of hemoparasites. However, the result came out negative which may be due to lack of optimization of the protocol or possibility of new species of these hemoparasites obtained. High detection of Plasmodium sp. and moderate detection on Leucocytozoon sp. which may influence by several common factors; high presence of vectors such as Aedes sp. which are typical in the tropical climate of Malaysia and lack of proper management against parasitic infections in quail’s production. In quail production, it applied basic principle of a good biosecurity practice without usage of drug and vaccine to prevent or treat the diseases based on quails being resistant to common poultry diseases which also included the haemoparasites. Nevertheless, avian haemoparasites can still be pathogenic to domestic poultry causing high mortalities, reproductive failure, retardation of growth and reduced productivity. Thus, this present study would serve as a reference point for further study and a pin-point to start emphasize on effect of diseases; hemoparasitic infections in quails and improve management of quails in controlling the disease. REFERENCES Mohammed, B. R., & Ejiofor, C. (2015). The Prospects and Limitations of Japanese Quail (Coturnix

coturnix japonica) Production in Nigeria. International Journal of Multidisciplinary and Current Research, 3, 920-926

Friend, M., Franson, J. C., Ciganovich, E. A., & Geological Survey (U.S.). (1999). Field manual of wildlife diseases: General field procedures and diseases of birds. Washington, DC: U.S. Dept. of the Interior, U.S. Geological Survey.

Hellgren, O., Waldenström, J., & Bensch, S. (2004). A new pcr assay for simultaneous studies of Leucocytozoon, Plasmodium, and Haemoproteus from avian blood. Journal of Parasitology, 90(4), 797-802. doi:10.1645/ge-184r1

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PATHOGENICITY AND PROPAGATION OF UPMT27 FIELD ISOLATE OF MALAYSIA FOWL ADENOVIRUS IN CHICKEN EMBRYO LIVER CELLS

A. SALISU1,4, I. NURULFIZA1,3,*, A.R. MARIATULQABTIAH1,3, M. HAIR-BEJO2,3,

A.R. OMAR2,3 AND I. AINI2,3 1Faculty of Biotechnology and Biomolecular Sciences, 2Faculty of Veterinary Medicine,

3Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor 4Department of Science Laboratory Technology, Jigawa State Polytechnic, Dutse,

Nigeria *Corresponding author: [email protected]

ABSTRACT

The poultry industry in Malaysia is fast growing but has become under threat due to inclusion bodies hepatitis (IBH) caused by strains of fowl adenovirus (FADV). The present study was carried out to propagate a field isolate of FAdV (UPM T27) and examine its pathogenicity in chicken egg embryo and chicken embryo liver cells. Five Specific Pathogen Free (SPF) chicken eggs were inoculated with 0.1mls of the FAdV UPMT27 isolate via chorioallantoic membrane (CAM) sac route. Supernatant from liver homogenate of dead embryo were used to infect freshly prepared chicken embryo liver (CEL) cells and observed for cytopathic effects for serial passages. Subsequently, passage 1 and 2 isolates were stained with H&E and observed under the microscope. Results show 100% mortality of the eggs at 11 days post inoculation and 100% CPE were observed at 4 dpi respectively. Eosinophilic inclusion bodies were also observed in the hepatocyte after H&E staining. These results confirmed that UPMT27 FAdV isolated from Malaysia are highly pathogenic and can be successfully propagated in CEL cells for future studies of the virus. Keywords: Propagation, Pathogenicity, Fowl Adenovirus, Chicken Embryo Liver Cell.

INTRODUCTION The global Poultry farming is affected by emerging and re-emerging of diseases and parasites, some of which are threat to certain types of birds (United Kingdom Department for enviroment food and rural affairs, 2012). Fowl adenovirus is a member of the family Adenoviridae, infecting chickens and responsible for sporadic disease condition, Inclusion body hepatitis (IBH) (Soumyalekshmi et al., 2014). It is an economically important disease in broilers chicken industry. Inclusion body hepatitis associated FAdV of serotype 8b was first reported in 2–5 weeks broiler chicks which was characterized by enlarged, mottled and friable liver with two to 30% intranuclear inclusion bodies in the hepatocytes. While many infections are sub clinical and appear to be of little economic or welfare importance, significant outbreaks of disease associated with adenovirus continue to occur. Cross-neutralization assay classified FAdV into five species and further divided into 12 serotypes whereglobal distribution and reported cases show the virus are frequently isolated both from healthy and affected chickens (McFerran and Smyth, 2000). In Malaysia, the first cases of IBH was isolated in 2005 from commercially broilers chickens with similar cases reported from neighboring states where high mortality and poor broiler performance were identified. Thus, cause significant impact on the poultry industry. However, the infection was diagnosed based on the histopathological lesions and detection of non- enveloped icosahedral adenovirus (Hair-Bejo, 2005). Propagation of the virus is an important phenomenon for the subsequent identification and typing of the virus. Previous findings reported the use


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of embryonated chicken eggs as a sensitive medium for the isolation and propagation of FAdVs (Soumyalekshmi. et al., 2014). Therefore, the aim of the present study was to propagate and examine the pathogenicity of FAdV UPMT27 isolate in chicken egg embryo and chicken embryo liver cells. MATERIAL AND METHODS The field isolate of FAdV UPM T-27 strain used was isolated from a broiler chicken, in Malaysia from samples that was submitted for the detection of FAdV to the Faculty of Veterinary Medicine, Universiti Putra Malaysia. The virus was inoculated into 9- day old SpecificPathogen Free chicken eggs and observed for mortality. Chorioallantoic membrane (CAM) and liver was then harvested from 100% cumulative mortality and homogenized in 1:2 PBS w/v (PH 7.4). The supernatant was inoculated into the 80% confluency of CEL cells, prepared from liver obtained from 13-15 days old SPF chicken egg embryos. The CEL cells was incubated at 370C and observed for 100% cytopathic effects (CPE) until passage 2. CEL cells were then seeded on cover slips in 6-well tissue culture plates and infected with the harvested Passage 1 and Passage 2 isolates, and further observed for 100% CPE. Four percent paraformaldehyde was used to inactivate the virus for 15 mins before subjected to H&E staining. The slips were then removed and inverted unto clean microscope slides and viewed under light microscope. RESULTS AND DISCUSSION Results show 80% and 100% mortality of the SPF eggs inoculated with the FAdV on day 3 and 11 post inoculation (dpi) respectively, which explored that the isolate is highly pathogenic, which is capable of producing heavy mortality in poultry farms. The liver was inflamed, friable with spots of haemorrhages, pale and shows 100 CPE within 4 dpi. Similarly, CEL cells was observed to produce theCPE from the 1st passage onward and were in the formed of clumping, rounding and detachment of cells. There were no observed changes in the control cells. The finding showed that this isolate is capable of inflicting serious damages to the liver of infected chicken. In H&E staining, Eosinophilic inclusion bodies in the hepatocytes of the infected cells were also observed. Thus, a typical histopathological lesion associated with FAdV infection were identified. REFERENCES Balamurugan, V. and Kataria, J.M. (2014). The Hydropericardium Syndrome in Poultry a Current

Division of Avian Diseases, Indian Veterinary Research Institute, Izatnagar, Bareilly, UP 243 122. India. Vet Res Commun pp 127-48

Hair-Bejo, M (2005). Inclusion Body Hepatitis in a Flock of Commercial Broilers Chickens. Journal of Veterinary Malaysia, 31: Pp-23-26.

McFerran, J.B. and Smyth, J.A. (2000): Avian Adenoviruses. Rev. Sci. Tech. 19: Pp-589-601. Soumyalekshmi, S., M. K., Ajith and Meshram, C. (2014). Isolation of Fowl Adenovirus in Chicken

Embryo Liver Cell Culture and its Detection by Hexon gene-based PCR. Ind. J. Sci. Res. and Tech. 2(3): Pp-33-36

United Kingdom. (2011). Development for Enviroment Food and Rural Affairs

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EFFECTS OF FEEDING DIFFERENT POSTBIOTICS ON GROWTH PERFORMANCE OF BROILERS UNDER HEAT STRESS

A.M. HUMAM1,5, T.C. LOH1,2*, H.L. FOO3, S.A. ASMARA1, M.I. ALSHELMANI2 AND

M.M. NOORDIN4 1Faculty of Agriculture, 5Animal resources department, Faculty of Agriculture,

University of Baghdad, Iraq. 2 Institute of Tropical Agriculture and Food Security, 3Faculty of Biotechnology and Biomolecular Sciences, 4Faculty of Veterinary Medicine,

Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. *Corresponding author: [email protected]

ABSTRACT

A feeding trial was conducted to investigate the effects of different postbiotics produced by Lactobacillus plantarum on growth performance of broiler under heat stress. A total of 252 broiler chicken were allocated into 6 dietary treatments: (1) negative control (free antibiotic); (2) positive control (0.02% antibiotic of oxytetracycline); (3) 0.02% ascorbic acid; (4) 0.3% postbiotic RI11; (5) 0.3% postbiotic RS5; and (6) 0.3% postbiotic UL4. Treatments were started in finisher diet at 21 day-olds until 42 day-olds with acute heat stress (36 ± 1°C) for 3 hours per day. The results showed that birds fed postbiotic (RI11) had higher (P < 0.05) final body weight, total weight gain and best FCR compared with other treatments. However, a total feed intake (FI) were not different (P>0.05) between all the treatments. The results obtained in this study showed that feeding of 0.3% postbiotic RI11 could improve the growth performance in broilers under heat stress. Keywords: postbiotic, growth performance, heat stress, broiler.

INTRODUCTION For several decades, feeding antibiotics to livestock for improving growth rate and food efficiency has been practiced (Choe et al., 2013). There is some evidence which suggests that antibiotics may also counteract the adverse consequences of stress responses such as cold and hot stress (Zulkifli et al., 2000). On the other hand, there is growing concern regarding the development of antibiotic-resistant bacteria which attributed to continuous feeding of antibiotics at sub-therapeutic levels. Furthermore, the possible antibiotic residue effects on animal production and health (Shazali et al. 2014). Because of these consequences, there is increasing public awareness and pressure to search for alternatives to antibiotics (Loh et al., 2014). Recently, postbiotics were used as a feed additive in poultry industries to promote the immune response and the performance of birds. Postbiotics are substances produced in the final or intermediate stage of metabolic process in Lactic acid bacteria. Various findings have reported that postbiotic improved the growth of animals when used as additive in broiler diet (Kareem et al., 2016). Various studies have been conducted to test the efficacy of postbiotics. However, no study has been conducted to examine the postbiotic in the broiler diets under heat stress. Thus, the present study objective was conducted to determine the effects of feeding different postbiotics on growth performance of broiler under heat stress. MATERIALS AND METHODS Experimental Design and Samples collection A total of 252, one-day-old (cobb500) chicks were obtained from a commercial hatchery. The chicks were randomly allotted into 6 treatment groups. Each group


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was contained 7 replicates, and 6 birds per replicate. The treatment groups included T1= basal diet (negative control), T2=basal diet + 0.02% oxytetracycline (positive control). T3=basal diet + 0.02% ascorbic acid, T4, T5 and T6=Basal diet + 0.3% postbiotic RI11, RS5 and UL4, respectively. An ambient temperature of 32 ± 1°C on d 1 was gradually reduced to 24 ± 1°C by d 21. The heat stress temperature was 36 ± 1°C for 3 hours per day from 21 to 42 day-olds. Feed and water were provided for ad libitum consumption. Birds were fed standard broiler starter crumble (3,069 kcal of ME/ kg; 22.1% CP) and finisher diet (3,140 kcal of ME/kg; 19.6% CP) from d 1 to 20 and d 21 to 42, respectively. Birds were offered on the same feed in the starter diet without treatment, and the treatments were started with finisher diet from 21 to 42 day-olds. Body weight (BW) and feed intake (FI) were measured weekly. These data were used to calculate body weight gain (BWG) and feed conversion ratio (FCR). RESULTS AND DISCUSSION The results of growth performance were significant differences between the groups. The final body weight, total weight gain and FCR of chickens supplemented with 0.3% RI11 (T4) were the highest (P<0.05) among the treatments, while the -ve control (T1) and +ve control (T2) showed the lowest FCR compared with the other groups. However, total feed intake (FI) were not different (P>0.05) between all the groups. Postbiotics produced by L. plantarum improved significantly the growth performance of broiler chickens. This effect could be explained by feeding of postbiotics containing antimicrobial and antioxidant substances produced by L. plantarum that provide nutrients and enhance biophysiological activities in the gut of animals. This would have resulted in an improved absorption and intestinal morphology of the animals (Thanh et al. 2009). On the other hand, a similar result was obtained for growth performance in broiler chicken and piglets when fed with 0.3% postbiotic (Kareem et al., 2016). REFERENCES Choe, D., Foo, H., Loh, T., Hair-Bejo, M., & Awis, Q. (2013). Inhibitory property of metabolite

combinations produced from lactobacillus plantarum strains. Pertanika Journal of Tropical Agricultural Science, 36, 79-87.

Kareem, K.Y., Loh, T.C., Foo, H.L., Asmara, S.A., Akit, H. (2016.) Effects of Dietary Postbiotic and Inulin on Growth Performance, IGF1 and GHR mRNA Expression, Faecal Microbiota and Volatile Fatty Acids in Broilers. BMC Veterinary Research 12(1): 163.

Loh, T. C., Choe, D. W., Foo, H. L., Sazili, A. Q., & Bejo, M. H. (2014). Effects of feeding different postbiotic metabolite combinations produced by Lactobacillus plantarum strains on egg quality and production performance, faecal parameters and plasma cholesterol in laying hens. BMC Veterinary Research, 10(1):149.

Shazali, N., Foo, H. L., Loh, T. C., Choe, D. W., & Abdul Rahim, R. (2014). Prevalence of antibiotic resistance in lactic acid bacteria isolated from the faeces of broiler chicken in Malaysia. Gut pathogens, 6 (1), 1

Thanh, N., Loh, T. C., Foo, H. L., Hair-Bejo, M., & Azhar, B. (2009). Effects of feeding metabolite combinations produced by Lactobacillus plantarum on growth performance, faecal microbial population, small intestine villus height and faecal volatile fatty acids in broilers. British poultry science, 50, 298-306.

Zulkifli, I., N. Abdullah, N. Mohd. Azrin, and Y.W. Ho. 2000. Growth Performance and Immune Response of Two Commercial Broiler Strains Fed Diets Containing Lactobacillus Cultures and Oxytetracycline under Heat Stress Conditions. British Poultry Science 41(5): 593–97.

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A STUDY OF INTERNAL PARASITES IN FREE RANGE TURKEYS IN SIMPANG RENGGAM FARM, JOHOR, MALAYSIA

P.A. SHIVANI AND I.H. LOKMAN*

Department of Veterinary Preclinical Sciences, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia


ABSTRACT Free range rearing is an integral part of turkey farming in most of the developing countries of world. Turkeys raised in free-range system are free to live and browse outside. Very scarce reports regarding internal parasites on free ranging turkeys are available from Malaysia. The present research project was designed to study internal parasites in free range turkeys in Malaysia from Simpang Renggam Farm, Kluang District, and Johor. The blood parasites found in turkeys were Leucocytozoon spp and Plasmodium spp at 1 (20%) and 2 (40%).Three species of fecal oocyst/egg infected the turkeys. Eimeria spp had the highest prevalence of 3(60%) followed by Heterakis gallinarum 2(40%) and Capillaria sp 1(20%). For the gastrointestinal worms, Capillaria spp worm were found at 20% and Acanthocephalan worm was found at 2(40%).Generally, of these 5 female birds, 2 (40%) were infected with blood parasites and 4(80%) were infected with gastrointestinal parasites. The study is second in its nature in providing the valuable information regarding endoparasites in turkeys from Malaysia. This information will essentially be helpful for the researchers and local veterinarians to develop strategies for both treatment and control of these endoparasites affecting turkeys. Keywords: turkey, free range, internal parasites, Malaysia.

INTRODUCTION The poultry industry provides meat that is preferred by almost all cultures, is affordable and of good quality. Turkey and chicken productions make up the main component of the commercial poultry. Global turkey meat production is about 5.7 m tonnes/year (Poultry Hub, 2018) Turkey meat is less consumed in Malaysia compared to western countries such as Europe and United States. Normally, turkey meat is sold at RM25 to RM30 per kg on average in some poultry farm (Suhaila and Khadijah, 2017). In addition, the lack of awareness makes turkey meat not popular as a delicacy or dish. Turkey parasites commonly seen include protozoa, helminths and arthropods. Parasitism causes tremendous loss in poultry production. whereby it decreases the growth rate and increases mortality. It is obvious that poultry maintained under free-range conditions may be heavily parasitized due to the fact that they scavenge in the farm area. It was the objective of this study to identify internal parasites harboured by free-range turkeys in Malaysia (Simpang Renggam Farm). MATERIALS AND METHODS Free-range female turkeys (5) were randomly chosen from the Simpang Renggam farm in order to collect the blood and faecal samples. All fowls were slaughtered using Halal – slaughtering method and the gastrointestinal tract was collected. Thin blood smears were made and stained with Giemsa stain to detect blood parasites under the microscope. Each fecal sample was analysed using the McMaster technique to identify quantitatively the helminthic eggs and coccidia oocysts that were possibly present. The intestines were separately cut open using a dissecting

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Myoris scissors and soaked in normal saline. The worms were collected and preserved in 70% alcohol. Then worms were directly examined and observed under a stereomicroscope. For confirmation, the worms were directly mounted with Lactophenol medium and viewed under compound microscope. In order to obtain results of parasite occurrence, prevalence was calculated. As stated by Thrusfield (2005), prevalence, P, is the number of instances of disease or related attributes (infection or presence of antibodies) in a known population, at a designated time, without distinction between old and new cases. RESULTS AND DISCUSSION Internal parasites constitute a major impediment to efficient poultry production including turkeys, thereby leading to substantial economic losses. Turkeys examined in this study were infected with Plasmodium spp (40%) and Leucocytozoon sp (20%). Out of all the internal parasites found, coccidian oocyst had the highest percentage. Eimeria spp had the occurrence of 80%. In keeping with the research done in Malaysia, oocyst of Eimeria spp. recorded the highest faecal egg count with 7300 epg (Suhaila and Khadijah, 2017). The fowls examined were infected with the nematodes, Capillaria spp and Heterakis gallinarum. This could be due to the reason that nematodes establish themselves in their hosts faster than the others trematodes and cestodes (Hoghoghi et al., 2014). Eggs (40%) and worms (20%) of Heterakis gallinarum were found from the faeces and intestinal tract samples. Capillaria spp (20%) eggs were only detected in the faeces. Endoparasite screening of free range turkeys done in Johor, Malaysia detected Capillaria spp. with 1200 epg. (Suhaila and Khadijah, 2017) Thorny-headed worms or Acanthocephalan was found in two out of five turkeys (40%) in this study. None of the studies done on turkeys in Malaysia stated the presence of Acanthocephalan. The occurrence of blood parasites of turkeys in this study was 40%. The overall high occurrence rate of 80% of gastrointestinal parasites in turkeys suggest that there are favourable environmental conditions around the farm, which are necessary for coccidian oocyst/helminth eggs survival and high reproductive efficiency of these parasites. Free range and scavenging behaviour of turkey’s increases exposure to soil and intermediate host, which may lead to parasitic infection. Therefore, turkeys raised in free-range condition of breeding should have a boundary and food served to them to reduce their search for food, which exposes them to intermediate hosts of parasites.

REFERENCES Hoghoghi Rad, N., Ramezani, A., Babazadeh, D., Falah, S., & Ghavami, S. (2014). Evaluation of

Gastrointestinal Helminths of Native Turkeys in Amol, Iran. J. World’s Poult. Res. Journal Homepage: J. World’s Poult. Res, 4(44), 86–88.

Opara, M. N., Osowa, D. K., & Maxwell, J. A. (2014). Blood and Gastrointestinal Parasites of Chickens and Turkeys Reared in the Tropical Rainforest Zone of Southeastern Nigeria. Open Journal of Veterinary Medicine, 4(12), 308–313.

Poultry Hub. (2018). Turkey. Retrieved January 26, 2018, from http://www.poultryhub.org/species/commercial-poultry/turkey/

PoultryDVM. (2018). Capillariasis in Chickens. Retrieved January 27, 2018, from http://www.poultrydvm.com/condition/capillaria-worms

Suhaila, A. H., & Khadijah, S. (2017). Parasites Prevalence In Poultry: Focusing On Free Range Turkeys (Meleagris gallopavo), 1–9.

http://www.poultryhub.org/species/commercial-poultry/turkey/

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UPREGULATION OF CHOLESTEROL STORAGE SIGNALLING GENES IN HEPG2 CELLS SUPPLEMENTED WITH EDIBLE BIRD NEST (EBN) EXTRACT

D. TAWAI, M.A.M. NOOR, F. SALLEH, I.S. RAZAK, R. MANSOR, A. IDERIS AND

M. AJAT* Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang,

Selangor, Malaysia *Corresponding author: [email protected]

ABSTRACT

“Caviar of the East” or better known as Edible-Bird Nest (EBN) is a highly valuable product composed of solidified salivary secretion of a few insectivorous swiftlet species. Swiftlets falls under the family Apodidae and the two main genera with the highest commercial values are Aerodramus (echolating swiftlets) and Collocalia (non-echolating swiftlets). The objective of this study is to determine the qualitative genes expression level associated in lipid signalling pathways such as Acetyl-CoA acetyltransferase 2 (ACAT2), Diacylglycerol O-acyltransferase 2 (DGAT2), Low density lipoprotein receptor (LDLR), Sterol regulatory element binding transcription factor 2 (SREBP2), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), Proprotein Convertase Subtilisin/kexin type 9 (PCSK9) in HepG2 mammalian cell with or without supplementation of EBN extract and exogenous lipid. Raw EBN were manually cleaned, dried in 60°C oven and left overnight. In this project, EBN supernatant were added with acetone using 1:2 ratio and kept in -80°C for an hour. HepG2 mammalian cells were cultured in six wells plates and each wells were treated according to Base Control, Positive Control, Negative Control, Treatment 1, Treatment 2 and Treatment 3. In this study, there were significant differences in terms of genes expression especially in ACAT2 and LDLR genes with supplementation of EBN extract and exogenous lipid concentrate. Keywords: Edible bird nest, swiftlet, lipid metabolism, genes expression.

INTRODUCTION Aerodramus fuciphagus is the most commonly found swiftlet species in Malaysia, producing the premium grade white nest due to its composition: purely solidified salivary secretion with high concentrations of N-acetylneuraminic acid (sialic acid) and epidermal growth factor (EGF) (Marcone, 2005); Wong et al., 2013; Looi et al., 2017). It is believed that EBN offers abundant of medicinal and health-boosting properties however there are not much scientific reports to prove this. Lipid droplets (LDs) are the major intracellular organelles specializing in storage of neutral lipids such as triglycerides and sterol esters. Excessive accumulation of lipid in LDs or disruption in energy homeostasis will result to various metabolic diseases such as obesity, diabetes, atherosclerosis and fatty liver disease (Ference et al., 2017; Krahmer et al., (2013). In this study, we determine the expression level of lipid signalling pathways genes such as Acetyl-CoA Acetyltransferase (ACAT), Diacylglycerol O-acyltransferase 2 (DGAT2), Sterol regulatory element binding transcription factor 2 (SREBP2), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), Low density lipoprotein receptor (LDLR), Proprotein Convertase Subtilisin/kexin type 9 (PCSK9) in HepG2 mammalian cells after supplemented with exogenous lipid and treated with or without Edible Bird Nest (EBN) extract. MATERIALS AND METHODS Raw EBN were cleaned with distilled water and feathers were removed manually before being dried in an oven at 60°C overnight. Dried EBN were finely grounded and


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mixed with Phosphate Buffer Saline (PBS) subsequently subjected to stewing in a waterbath at 70°C for 5 hours. EBN supernatant were collected and subjected to acetone protein precipitation. EBN extract was filtered through 0.22 µm filter to get a pure EBN extract. HEPG2 mammalian cells derived from liver tissue were used in this study. Cells were cultured in completed medium supplemented exogenous lipids and cholesterol with or without EBN extract. Simvastatin was used as positive control. RNA was extracted from cells according to supplier manual and were subject to RT-PCR. RESULTS AND DISCUSSION Among the six genes related to lipid signalling pathways tested, EBN extract have been found to have significant effects on ACAT2 and LDLR genes. ACAT2 gene is responsible for the synthesis of sterol ester where it will be incorporated in lipid droplets for lipid storage whereas LDLR gene is the most important key player in internalization of LDL from the blood stream into the cells. Hence, upregulation of LDLR gene will increase the number of LDL receptors on the outer surface of the cells thus reducing the amount of circulating LDL (bad cholesterol) in the blood which is very beneficial so as to lower the risk of atherosclerosis or cholesterol deposition in the vessels. As for PCSK9 gene, result remains inconclusive for now. However, EBN extract supplementation may have a positive effect in reducing PCSK9 expression. Further research to look into significant of EBN extract towards lipid signaling pathways genes expression via quantitative method in the future will be conducted. REFERENCES Ference, B. A, Ginsberg, H.N., Graham, I., Ray, K.K., Packard, C.J., Catapano, A.L., (2017). Low-density

lipoproteins cause atherosclerotic cardiovascular disease. Evidence from genetic, epidemiologic, and clinical studies. A consensus statement from the European Atherosclerosis Society Consensus Panel, European Heart Journal, 38:32(21): 2459–2472.

Krahmer, R.V. Farese Jr., T.C. Walther, (2013). Balancing the fat: lipid droplets and human disease, EMBO Mol. Med. 5 973–983.

Looi, Q. H., Amin H., Aini, I., Zuki, M., Omar, A. R. (2017). De novo transcriptome analysis shows differential expression of genes in salivary glands of edible bird’s nest producing swiftlet. BMC Genomics. 18, 504

Marcone M.F. (2005). Characterization of the edible bird’s nests the “Caviar of the East”. Food Res Int: 38(10): 1125– 1134. Wong, R. S. Y. (2013) Edible Bird's Nest: Food or Medicine? Chinese Journal of Integrative Medicine,

19, 643-649.

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FEEDING PAOB AT DIFFERENT LEVEL OF AMINO ACIDS IN THE DIET OF BROILER CHICKENS

C.H. GOH1, T.C. LOH1,2,* AND H.L. FOO3,4

1Department of Animal Science, Faculty of Agriculture; 2Institute of Tropical Agriculture, 3Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences; 4Institute of Biological Science; Universiti Putra Malaysia,

43400 UPM Serdang, Selangor, Malaysia *Corresponding author: [email protected]

ABSTRACT

The purpose of this study was to investigate the effect of dietary supplementation with organic acids (OA) with different level of amino acids on the growth performance, digestibility and microflora count in broiler chickens. The diet supplemented with organic acid and palm fat (POAB) in broiler diet showed an increase in body weight gain (2963.00 g) compared to control group (2694.55 g) at the end of 42 day of experiment. A similar trend was found for crude fat, methionine and lysine digestibility. It showed that the OAPF able to improve the nutrients digestion of the chickens. From the results, the OAPF able to improve the growth performance by increase the digestibility of the chickens. Moreover, the treatment groups supplemented with OAPF promote the growth of beneficial bacteria which was indicated by higher lactic acid bacteria (LAB) count compared to the treatment without organic acids. Keywords: Organic acids, broiler, growth performance, digestibility, microflora count.

INTRODUCTION Antibiotics are commonly used in poultry diets in order to prevent diseases and to improve the performance. However, there has been growing public concern about the risk of bacterial resistance associated with the routine use of Antibiotic Growth Promoter (AGP) in livestock feeds. The organic acid mixtures might be more efficient than some antibiotic growth promoter in improving broiler performance. Organic acids are natural constituents of plant and animal tissues and their use as feed additives is now being studied worldwide to replace antibiotics. A wide range of organic acids with variable physical and chemical properties are available for poultry, of which many are used in the drinking water or mixed with the feed. Besides that, organic acids also contributed greatly to the profit in poultry production as it provides consumer with health and nutritious poultry products (Adil et al., 2010). The mode of action of organic acids in animal diets has not been clearly clarified; this inadequate understanding has limited the application of organic acids in broiler diets. However, several possible mechanisms have been proposed and most of them have been associated with: (1) reduced pH in diets and subsequent reduction of the pH in the GIT, (2) better nutrient utilization in diets by increasing nutrient retention, and (3) inhibition of pathogenic bacterial growth (Mroz, 2005). MATERIALS AND METHODS A total of 384-day-old chickens were used in this study. This experiment will be conducted in a poultry research unit, UPM. The chickens were randomly assigned into 8 treatment groups with 6 replicates per treatment. Each experimental unit was consisted of 8 chickens. Starter diet was feed from day 0 to day 21, while grower diet was feed from day 21 to day 42. The POAB used in this study was supplied by Sunzen Lifescience Sdn Bhd. The chickens were allocated to the following treatments: For


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starter diet, T1: 21% Crude protein (CP) (Lys 1.18%; Met 0.45%), T2: 21% CP (Lys 1.0%; Met 0.45%), T3: 21% CP (Lys 1.18%; Met 0.4%), T4: 21% CP (Lys 1.0%; Met 0.4%), T5: 21% CP (Lys 1.18%; Met 0.45%) + POAB, T6: 21% CP (Lys 1.0%; Met 0.45%) + POAB, T7: 21% CP (Lys 1.18%; Met 0.4%) + POAB, T8: 21% CP (Lys 1.0%; Met 0.4%) + POAB. For grower diet, T1: 19% CP (Lys 0.95%; Met 0.39%), T2: 19% CP (Lys 0.75%; Met 0.39%), T3: 19% CP (Lys 0.95%; Met 0.25%), T4: 19% CP (Lys 0.75%; Met 0.25%), T5: 19% CP (Lys 0.95%; Met 0.39%) + POAB, T6: 19% CP (Lys 0.75%; Met 0.39%) + POAB, T7: 19% CP (Lys 0.95%; Met 0.25%) + POAB, T8: 19% CP (Lys 0.75%; Met 0.25%) + POAB.The experiment was conducted for 42 days. The individual body weight (BW) and cage feed intake (FI) were recorded weekly and live weight gain (WG), average daily weight gain (ADG) and feed conversion ratio (FCR) were calculated. Ileal digesta samples were taken for nutrient analysis and apparent digestibility measurement. RESULTS AND DISCUSSION This result showed that improved growth performance was dependent on the OAPF. Besides that, it was also affected by the reduced level of amino acids in the diets. The lower inclusion of lysine and methionine caused the decreased of BWG of the chickens. However, the inclusion of OAPF in the diets improved the BWG of the chickens although the AA levels were reduced in the diets. There was no significant difference among the treatment groups during the initial body weight. The treatment group supplemented with POAB had a significant higher performance. Most of the treatment groups with POAB also showed a significant lower FCR compared to the treatment group without supplementation of POAB. The improvement in the FCR could be possibly due to better utilization of nutrients resulting in increased body weight gain (Adil et al. 2010). Moreover, the dietary supplementation of organic acids increased the body weight and FCR in broiler chicken (Fascina et al. 2012). Treatment groups supplemented with POAB clearly showed a better digestibility to the treatment without POAB. Besides that, Garciá et al. (2007) reported that supplementation of formic acid (0.5% or 1.0%) in broiler finisher diet was found to improve apparent ileal digestibility (AID) of CP (72.5% or 73.5%, respectively) as compared with control (60.7% CP). From the result, it showed that the supplementation of POAB able to increase the LAB and reduced ENT cell population. Decreased faecal pH values may also affected the decreased of pathogenic bacteria in broiler excreta. However, reduction of amino acids of the feed had no effect on the microbial population in the faecal samples. REFERENCES Adil, S., Tufail, B., Gulam, A.B., Masood, S. and Manzoor, R. (2010). Effect of dietary supplementation

of organic acids on performance, intestinal histomorphology, and serum biochemistry of broiler chicken. Vet Med Int:1–7.

Fascina, V.B., Sartori, J.R., Gonzales, E., Barros De Carvalho, F., Pereira De Souza, I.M.G., Polycarpo, G.V., Stradiotti, A.C. and Pelícia, V.C. (2012). Phytogenic additives and organic acids in broiler chicken diets. Revista Brasileira de Zootecnia. 41(10):2189–2197.

Garciá, V., Catalá-Gregori, P., HernáNdez, F., Megiás, M.D. and Madrid, J. (2007). Effect of formic acid and plant extracts on growth, nutrient digestibility, intestine mucosa morphology, and meat yield of broilers. J Appl Poultry Res. 16:555–562.

Mroz, Z. (2005). Organic acids as potential alternatives to antibiotic growth promoters for pigs. Adv Pork Prod; 16:169-182.

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MYCOFLORA AND NATURAL OCCURRANCE MYCOTOXINS IN CORN, SOYBEAN MEAL AND WHEAT POLLARD IN MALAYSIA

N. SHAZALI1, T.C. LOH1,4,*, H.L. FOO2,3, H. AKIT1 AND M.H. KAMALLUDIN1

1Department of Animal Science, Faculty of Agriculture; 2Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences; 3 Institute of

Biological Science; 4Institute of Tropical Agriculture, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia


ABSTRACT

The purpose of this study was to identify the mycoflora and mycotoxins production in corn, soybean meal and wheat pollard in Malaysia. The study demonstrated that corn had significantly higher mycoflora counts as compared to other grains. Corn, wheat pollard and soybean meal contained at least one of the main mycotoxigenic genera such as Aspergillus spp., Penicillium spp., and Fusarium spp. and the most toxicogenic fungi (15.4% to 75%). All raw materials were contaminated with fungus and mycotoxins have been detected with a various concentration of aflatoxins (AF), ochratoxin A (OTA), zearalenone (ZEA), and deoxynivalenol (DON). Overall results show that corn, soybean meal and wheat pollard contained a higher concentration of AFB2, DON and ZEA contamination. As conclusion, the presence of fungi and mycotoxins indicates the existence of contamination. Keywords: Mycoflora, mycotoxins, corn, soybean meal, wheat pollard.

INTRODUCTION Animal feed plays an important part in the food chain and has implications for the composition and quality of the livestock products that people consume. However, there will be undesirable substances in the animal feed that commonly known as contaminants. Mycotoxins were first mentioned in the early 1960s when the discovery of the aflatoxins was made (Richard, 2007). These metabolites are produced from mycotoxin-producing fungi such as Aspergillus, Penicillium, and Fusarium, and of which thousands are known to be toxic to animals and humans (Ismaiel and Papenbrock 2015). The implication of mycotoxins lies on the appearance of several toxicosis and diseases in both humans and animals, especially in monogastrics (Hussein and Brasel, 2001). Mycotoxins also affect prior, during, and post-harvesting, transportation and storage of a wide variety of agricultural products, which is reflected in condemned agricultural products and important agro-economic losses (Zain, 2011). Hence as part of a proper mycotoxin risk management, surveying the mycotoxin occurrence is very important to allow feed and animal producers to assess the risk of using certain feed ingredients or feeds from different regions. Under practical conditions, no poultry feed is completely free of mycotoxins and it's indicating a clear and persistent danger (Bryden, 2012). Additionally, no feed can be expected to contain only one mycotoxin (Broom, 2015). Thus, the purpose of this study was to identify the mycoflora and mycotoxins production in corn, soybean meal and wheat pollard in Malaysia. MATERIALS AND METHODS Mycological and mycotoxins assessment were determined by using a minimal modification based on Pitt and Hocking method (2009). The grains were obtained commercially from a local company in Selangor. The enumeration was performed in


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two way which is isolation frequency and colony form unit (CFU/g dry weight). Then pure colonies were cultured in Potato Dextrose Agar and kept in 4 ºC for fungal identification. Fungal identification was carried out based on fungus morphology. Furthermore, 25 g of the ground samples were used and blended at high speed (Waring, FL, USA) and filtered through fluted filter paper (Vicam, Germany). Volume of 10 mL of filtered diluted extract was passed through Aflatoxin, DON and Fumonisin Test immunoaffinity column (Vicam, Germany). The eluents in the column was collected by passing 1 mL of HPLC grade methanol through the column and adding 1 mL purified water to the elute. Total mycotoxins occurrence was later determined by using UHPLC-MS. RESULTS AND DISCUSSIONS The total fungal population isolated from corn, wheat pollard and soybean meal show a wide range of mycoflora counts and all the analysed samples had counted over the proposed limits of 1×104 CFU/g−1(GMP, 2006). However, corn had significantly higher mycoflora counts as compared to other grains. Corn, wheat pollard and soybean meal samples contained at least one of the main mycotoxigenic genera such as Aspergillus spp., Penicillium spp., and Fusarium spp. and the most toxicogenic fungi (15.4% to 75%). Aspergillus flavus and Aspergillus niger were isolated from all raw materials. Aspergillus niger was isolated from raw materials at levels media from 25% and Aspergillus flavus from 41.7%, respectively. The relative density of isolated Penicillium spp. was occurring from 21% to 47% of total samples isolated. The relative density of isolated Fusarium spp. from the raw materials was occurring from 25% to 58.3% of the total samples and Fusarium verticillioides was found as predominant in corn and soybean meal followed by Fusarium subglutinans in wheat pollard. All raw materials were contaminated with fungus and mycotoxins have been detected with a various concentration of aflatoxins B1 (AFB1), aflatoxins B2 (AFB2), aflatoxins G1 (AFG1), aflatoxins G2 (AFG2), ochratoxin A (OTA), zearalenone (ZEA), and deoxynivalenol (DON). Overall results show that corn, soybean meal and wheat pollard contained a higher concentration of AFB2, DON and ZEA contamination. AFB1 (1.80 µg/ml) was detected in soybean meal and the concentration was above the recommended limits as compared to other grains. Soybean meal had a higher concentration of AFB2 (452.93 µg/ml) then followed by corn and wheat pollard. Soybean meal contained higher levels of AFG1 (121.94 µg/ml) than other raw materials. Wheat pollard contained a lower concentration of AFG2 (4.67 µg/ml) as compared to corn and soybean meal. All raw materials had a similar concentration of DON, OTA and ZEA. As conclusion, the presence of fungi and mycotoxins indicates the existence of contamination. REFERENCES Broom, L. (2015). Mycotoxins and the intestine. Animal Nutrition. 1: 262-265. Bryden, W.L. (2012). Mycotoxin contamination of the feed supply chain: Implications for animal

productivity and feed security. Animal Feed Science and Technology, 173: 134-158. Hussein, H.S. and Brasel, J.M. (2001). Toxicity, metabolism, and impact of mycotoxins on humans and

animals. Toxicology, 167: 101-134. Ismaiel, A. A. and Papenbrock, J. (2015). Mycotoxins: Producing Fungi and Mechanisms of

Phytotoxicity. Agriculture, 5: 492-537. Pitt, J.I. and Hocking, A.D. (2009). Fungi and Food Spoilage. Springer, Dordrecht, Heidelberg, London,

New York.

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SALMONELLA ENTERICA SEROVAR ENTERITIDIS IN MEAT PRODUCTS

E. KHOO*, H.W. HO, M.N. NORAZARIYAH, R.S.N. HANANI, M. NAFIZAH, M.A. NOORMAH, R. SAIFU NAZRI, Y. ROSNAH, M. FHITRI, R. ROSELIZA AND

A.R. SOHAYATI Veterinary Research Institute, 59 Jalan Sultan Azlan Shah,

31400 Ipoh, Perak, Malaysia. *Corresponding author: [email protected]

ABSTRACT

Salmonella enterica serovar Enteritidis is the most common serovar identified in both human and animals worldwide since 1993 until now. This study reported the positive isolation of S. Enteritidis in various meat products using 10 years serotyping data in Department of Veterinary Services (DVS) Malaysia. From 2008 to 2017, a total of 8251 Salmonella isolates were received for further serotype. On average, the percentage of positive S. Enteritidis were about 19.5% (1611/8251) and were found to be predominant in chicken meats. From 2008 to 2017, the percentage of positive cases ranged from 7% to 32%. These passive surveillance data might not represent the true risk of S. Enteritidis in Malaysia as the transmission of S. Enteritidis infections is complex due to the fact that contamination can occur at multiple stages from farms and along food supply chain includes processing, distribution, retailing and handling by consumers. DVS Malaysia implemented a control program on Salmonellosis through annual National Surveillance Program and certification scheme such as Malaysia Good Agricultural Practices (MyGAP). The data from the program was shared with Ministry of Health (MOH) for better control strategy at each designated approved authority to ensure food safety for public consumption.

Keywords: S. enterica ser. Enteritidis, animals, livestock products, serotyping. INTRODUCTION Salmonella is one of the most common foodborne pathogen that can causes gastroenteritis, bacteremia and other complications of non-typhoidal Salmonellosis in human. In Veterinary Research Institute (VRI) Annual Report, well documented Salmonella outbreak in animal dated back to ϭϵϳϬ’s with S. Blockley and S. Sofia infect poultry farms. In ϭϵϴϬ’s, cattle farms were infected with S. Dublin and S. Typhimurium, while poultry farm infected with S. Pullorum. Starting from ϭϵϵϬ’s, there is a totally increase in S. Enteritidis, where S. Dublin and S. Pullorum infection gain successful control with nationwide vaccination program. Since 1993 until now, S. Enteritidis is the most common circulating serovar identified in both human and animals (Pui et al., 2011; Shafini et al., 2017). Nevertheless, the Department of Veterinary Services (DVS) Malaysia had put in effort to control Salmonellosis where it is included in annual National Surveillance Program to ensure safe and quality meat products for public consumption. This study report on positive cases of S. Enteritidis from meat products collected from 10 years serotyping data (year 2008 to 2017) from Department of Veterinary Services (DVS) Malaysia. MATERIALS AND METHODS The Salmonella isolation and confirmation were carried out by the DVS Regional Veterinary Laboratories using conventional culture and biochemical tests. A total of 8251 Salmonella isolates were received for further serotype in Veterinary


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Research Institute (VRI, DVS) Malaysia between the study periods. Salmonella sp. were serotyped using slide agglutination test and Schworm agar method referring to the White-Kauffmann-Le Minor Scheme, 2007 using commercial specific somatic and flagellar antisera (OIE Terrestrial Manual, 2010).

RESULT AND DISCUSSION On average, the percentage of positive S. Enteritidis were about 19.5% (1611/8251) and were found to be predominant in chicken meats. From 2008 until 2010, the prevalence ranged from 17% to 22%. In year 2011, there was sudden increased up to 32% and reduce to 14% and 21% in year 2012 and 2013. In 2014 and 2015, the percentage was found to increase above 30% again and later greatly reduced to 10% and 7.3% in year 2016 and 2017. The positive isolation of Salmonella and S. Enteritidis from various meat products were tabulated in Table 1 and Table 2. Table 1: Distribution of Salmonella and S. Enteritidis isolated from year 2008 to 2017 in Malaysia. 2008 2009 2010 2011 2012 2013 2014 2015 2016 2017 Total

Total Salmonella isolates

718 784 717 725 1472 725 636 757 611 1106 8251

S. Enteritidis

159 146 126 232 210 156 195 242 64 81 1611

% S. Enteritidis

22.1 18.6 17.6 32.0 14.3 21.5 30.7 32.0 10.5 7.3 19.5

Table 2: Distribution of S. Enteritidis isolated from various meat products from year 2008 to 2017. Meat Type

2008 2009 2010 2011 2012 2013 2014 2015 2016 2017 Total

Chicken 152 145 118 226 178 143 185 239 63 79 1528

Buffalo 3 - 6 - 18 3 4 - 1 - 35

Cattle 1 1 - 5 2 8 3 3 - - 23

Goat - - - 1 11 1 2 - - - 15

Duck 3 - 1 - 1 - - - - 2 7

Pig - - 1 - - 1 1 - - - 3

Total 159 146 126 232 210 156 195 242 64 81 1611

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Although these passive surveillance data might not represent the true prevalence of S. Enteritidis in Malaysia, but it does provide information on number of positive isolation from the meat products over the years. Decreasing number of S. Enteritidis in recent 2 years highly due to other Salmonella serovar become predominant over S. Enteritidis, such as S. Brancaster, S. Corvallis and S. Albany (source from VRI unpublished data). The true prevalence of Salmonellosis is difficult to determine because it depends much on right sample size collected. The control and prevention of Salmonellosis in animal remain as a challenge to veterinarians because clinically healthy carrier animals shed the pathogen in feces and contaminate the environment. The persistence of Salmonella in farms or premises is difficult to eliminate as they are capable of surviving in varies environmental conditions and rely much on the hygienic or disinfection practices in place. The transmission of S. Enteritidis infections is complex due to the fact that contamination can occur at multiple stages from farms and along food supply chain includes processing, distribution, retailing and handling by consumers. DVS Malaysia has control program on Salmonellosis through annual National Surveillance Program and certification scheme such as Malaysia Good Agricultural Practices (MyGAP). Besides, guidelines to control Salmonellosis is written in Malaysia Veterinary Protocol (Document No: PVM 6(15):1/2011). DVS can work together with Ministry of Health (MOH) in terms of disease data sharing to come out with better control strategy at each designated approved authority to ensure food safety for public consumption. ACKNOWLEGDEMENTS The author would like to thank the Director General of Veterinary Services Malaysia (DVS) for his kind permission to publish this scientific paper. Special thanks to Bacteriology Section in all DVS Regional Veterinary Laboratories and staff of Bacteriology Section, Veterinary Research Institute (VRI, DVS) for their contribution in this study. REFERENCES OIE Terrestrial Manual, 2010. Chapter 2.9.9, Salmonellosis. Patrick A.D. Grimont and François-Xavier Weill, (2007). Antigenic Formulae of the Salmonella

Serovars. 9th edition. Paris, France: WHO Collaborating Centre for Reference and Research on Salmonella, Institut Pasteur.

Salmonellosis Unggas, Protokol Veterinar Malaysia (No. Dokumentasi: PVM 6(15):1/2011) Jabatan Perkhidmatan Veterinar, Kementerian Pertanian Dan Industri Asas Tani Malaysia.

Pui, C. F., Wong, W. C., Chai, L. C., Tunung, R., Jeyaletchumi, P., Noor Hidayah, M. S., Ubong, A., Farinazleen, M. G., Cheah, Y.K. and Son, R. (2011). Review Article Salmonella: A foodborne pathogen. International Food Research Journal 18: 465-473.

Quinn P., Carter M.E., Markey B.A. and Carter G.R. Clinical Veterinary Microbiology. London: Wolfe Publishing, 1994.

Shafini, A.B., Son, R., Mahyudin, N.A., Rukayadi, Y., and Tuan Zainazor, T.C. (2017). Prevalence of Salmonella spp. in chicken and beef from retail outlets in Malaysia. International Food Research Journal24(1):437-449.

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EPIDEMIOLOGICAL STUDY OF MYCOPLASMA GALLISEPTICUM AND MYCOPLASMA SYNOVIAE: CASES FROM LABORATORIES IN MALAYSIA FROM 2012- 2017

P.X. LIM AND N.M.A.N.M. FAIZ1,*

1Department of Veterinary Clinical Studies, Faculty of Veterinary Medicine Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia


ABSTRACT Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) are ubiquitous in major poultry producing countries. Both organisms can cause wide range of complications especially respiratory and joint problems that lead to significant economic losses to the poultry industry such as production losses, mortalities, condemnations and cost of medications. In this study, the objective was to analyse the epidemiology of MG and MS cases from data of four diagnostic laboratories in Malaysia. Bacteriology, serology and molecular detection data from samples submitted to these laboratories from 2012 to 2017 were collected. Results were analysed for laboratory tests, time trends, geographical distribution, age group and purpose of flock. In total, 54863 samples were submitted to the laboratories for analysis in the study period. The percentage of Mycoplasma sp. isolated via bacterial culture isolation is 7.5% from samples collected. Meanwhile, MG and MS data shows 54% and 28% from serology detection as well as 28% and 37% based on PCR tests respectively. MG and MS are increasing throughout the 6 years duration, and more often involving older age flock. The high number of Mycoplasma infection cases in poultry flocks in Malaysia warrants improved strategies on control and prevention in the future. Keywords: Mycoplasma gallisepticum, Mycoplasma synoviae, poultry, epidemiology, serology.

INTRODUCTION The Mollicutes or commonly known as the mycoplasmas, are the smallest known self-replicating prokaryotes surrounded by only plasma membrane as they do not possess cell walls (Bradbury, 2005). Generally, mycoplasmas are host-specific. Among the Mycoplasma species that infect avian species, M. gallisepticum (MG) and M. synoviae (MS) are considered the most important pathogenic mycoplasmas affecting poultry worldwide (OIE, 2008). MG predominantly causes respiratory distress clinical signs in poultry, while MS, also known as infectious synovitis can cause additional inflammation to joints. Diagnosis of mycoplasmas can be done via identification of the organism, detection of the antibodies, as well as detection of DNA of organism (Umar et al., 2016). Previous studies revealed a high prevalence of MG in Malaysia (Faisal et al., 2011) whereas the last study of MS in Malaysia is dated back to 2009 indicates MS is prevalent at low rate (Chuah, 2009). Hence this study is to update and analyse the epidemiology of MG and MS based on poultry samples submitted to various laboratories in the recent 6 years. MATERIALS AND METHODS Submission samples records to test for. MG and MS assay from year 2012 to 2017 were reviewed from 4 laboratories. The information was collected via computer software system ie the BioChek (BioChek B.V, The Netherlands) system, logbooks, submission forms and compiled records from all the labs. Criteria for the inclusion of study includes the date of samples, age of flock, breed of flock, purpose of the flock, geographical distribution of the poultry farms to where the samples originate, sample types, total samples, total positive and negative samples, types of diagnostic

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methods used, mean titre (for ELISA), reason of sample submission, vaccination status of farm, antibiotic sensitivity test if applicable. Data was then analysed, tabulated and transformed to charts via SPSS Statistics (IBM Corporation, New York) and Microsoft Excel (Microsoft Windows, Washington) based on types of diagnostic tests, time trends, geographical distribution, age of the flock, purpose of flock using the positive samples from the total samples submitted to all 4 laboratories. RESULTS AND DISCUSSION The total positive cases of Mycoplasma sp. out of the total samples submitted for diagnosis is 7.5%. On the other hand, out of the total samples collected from all labs that performed ELISA and PCR, positive cases for MG and MS were 54% and 28% as well as 50% and 37% respectively. The incidence of Mycoplasma sp. out of the total samples is lower via bacterial isolation method (7.5%) compared to ELISA (MG: 54%; MS: 50%) and PCR (MG: 50%; MS: 37%). Possible reasons could be due to the fastidious nature of Mycoplasma sp. Due to the long incubation period, many medias are overgrown by swarming bacteria. Furthermore, other contributing factors could be due to the presence of non-cultivable or dead pathogens, the lack of specific isolation media, condition of sample swab during transportation and type of sample swabs.

Time Trends MG and MS show a general increase in trend in the number of positive cases over the 6 years period. The lack of effective control programs and biosecurity measures could be the inciting cause of the increase.

Geographical distribution For MG, Kedah tops the overall positive cases from the total samples sent to laboratories with 96.34% frequency rate whereas Johor has the lowest with only 15.55%. For MS, Sabah is highest with 62% and lowest in Melaka (4%) among all samples submitted. The number of positive samples varies from farm to farm as each farm practises a different level of biosecurity and bio-surveillance.

Age Group For MG, the positive cases out of the total samples submitted for MG assay increases with age group starting from week old to the highest in > 20 weeks age group serologically whereas PCR results show the highest belonged to age group 9 – 20 weeks old. Serologically, this can possibly due to a complete vaccination regime in

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older age group, thus, higher seroconversion of vaccines to antibodies. The reason behind the PCR findings can be due to horizontal transmission of infected individual, or stress due to onset of lay and various health programs that are ongoing. For MS, the highest positive cases for both ELISA and PCR are found to be in the older flocks (>20 weeks old). It is supported by various studies that show high incidence of MS in layer flocks worldwide. It could be due to the multiple – age housing system in layer production and lower biosecurity standards in this field. Table: Serological and PCR results of MG in Chicken in relation to Age Group

Age Group Serology PCR

DOC 1470/4212 (34.90%) 12/27 (44.44%)

1-8 weeks 678/5024 (13.50%) 25/70 (35.71%)

9-20 weeks 2716/6008 (45.21%) 3/6 (50%)

>20 weeks 19584/29048 (67.42%) 14/50 (28%)

Purpose of Flock For MG, highest incidence is seen in layers, and least in breeders via PCR. The longer life span of layers increases the chances of field exposure to MG. However, although breeders share a long lifespan, more periodical prophylactic treatment involving the use of antibiotics and more intensive vaccination is practiced in breeder flocks compared to layers. MG vaccination is not practiced in some broiler farms in Malaysia, hence, sero-positivity of broiler towards MG is lowest. On the contrary, the comparable positive rate of MG infection among broilers (31.46%) with layers (31.65%) via PCR might signify the emergence of MG disease among broilers. For MS, PCR and ELISA results show a completely opposite trend whereby the highest MS infection based on PCR results is observed in layers, followed by broilers and breeders. Infection rate is more frequently observed in multi-age farm, which is usually practised in layers. Lowest infection rate with highest sero-positivity in breeders can be associated with the vaccination regime. The more frequently the vaccination was administered for other diseases, the lower the probability of flock infection with MS. Breeders usually have a more thorough vaccination regime than layers, hence, the higher immunity of breeders towards other diseases increases the immunity towards MS infection. Concurrent infection of MG and MS The concurrent infection of MG and MS is as high as 13% in this study. It is actually reported to be a common incident in other countries. CONCLUSION This study shows a high number of MG and MS infection in Malaysia poultry industry based on the total samples collected from all laboratories as stated. Higher cases are seen in older flocks. Increasing trends of MG and MS cases based on the laboratories

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samples received from year 2012 – 2017 warrants improvement in the control, prevention and eradication of this important poultry disease. REFERENCES Bradbury, J. M. (2005). Poultry mycoplasmas: Sophisticated pathogens in simple guise. British Poultry

Science, 46(2), 125–136. https://doi.org/10.1080/00071660500066282 Chuah, L. L. (2009). Prevalence of Myoplasma synoviae in commercial poultry farms in Malaysia. DVM

Thesis, Universiti Putra Malaysia. Faisal, Z., Ideris, A., Hair-Bejo, M., Omar, A. R., & ChingGiap, T. (2011). The prevalence of Mycoplasma

gallisepticum infection in chickens from Peninsular Malaysia. Journal of Animal and Veterinary Advances, 10(14), 1867–1874. https://doi.org/10.3923/javaa.2011.1867.1874

Umar, S., Munir, M. T., Ur-Rehman, Z., Subhan, S., Azam, T., & Shah, M. A. A. (2016). Mycoplasmosis in poultry: Update on diagnosis and preventive measures. World’s Poultry Science Journal, 73(1), 17–28. https://doi.org/10.1017/S0043933916000830

OIE (2008) Avian mycoplasmosis (Mycoplasma gallisepticum, Mycoplasma synoviae) In: Manual of diagnostic tests and vaccines for terrestrial animals, 2008, pp. 525-541.

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DETERMINATION OF NUTRIENT COMPOSITION AND PRESENCE OF BACTERIA IN RAW VERSUS BOILED CHICKEN LIVER DIET

K.N. TAN1, M.A.R.P. AZAZIAH1,*, H.A. HASSIM1 AND S. KHAIRANI-BEJO1

1Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia


ABSTRACT

The purpose of this study was to determine the nutrient composition and presence of bacteria in both raw and boiled chicken livers. A total of six samples for each raw and boiled chicken livers, were prepared for the nutrient composition analysis. Both samples were also prepared for bacteria isolation and identification. All samples were cultured on the blood agar and tryptic soy agar for bacteria isolation. The isolated bacteria were identified using biochemical tests. Result shows that all proximate analysis for moisture content, dry matter, crude fat and crude protein, were statistically difference between raw and boiled chicken liver, except for ash content percentage. The bacteria species isolated from raw chicken livers were Aeromonas species, Streptococcus species, Actinomyces species, Proteus mirabilis and Salmonella species. There was absence of viable bacteria isolated from boiled chicken livers. Thus, it can be concluded that the raw chicken liver contains higher nutrients composition but harbours numerous opportunistic bacteria that might compromise the general health in pets as well as to the public. Keywords: raw, boiled, chicken liver, proximate analysis, bacteria isolation.

INTRODUCTION There were more than 93.2% of dogs and 98.8% of cats consumed at least half of their food in the form of commercial pet foods (Laflamme et al., 2008). However, it is getting more popular among veterinarians and pet owners in providing home-prepared and raw food diets for the pets with the data of 30.4% of dogs and 13.1% of cats received non-commercial pet food (Michel et al., 2008). The preference of home-prepared and raw food diets with the usage of chicken livers for pet increases with the claims of various benefits, including the fact of chicken liver is a nutrient-rich organ and provides alternative sources of protein. However, food safety and public health are of concerns when some pets are fed with raw chicken liver diets due to the possibility of presence of various pathogenic organisms in the raw foods (Buff et al., 2014; Freeman et al., 2013). There are limited studies conducted about the pet feeding practices in Malaysia. Therefore, this study might serve as an important client education for the pet owners in terms of the food safety and nutritive values of home-prepared chicken liver diets for dogs. Hence, this study was aimed to determine the nutrient composition and presence of bacteria in both raw and boiled chicken livers. MATERIALS AND METHODS The fresh chicken livers were purchased from local wet market and further prepared for both raw and boiled chicken livers. A total of six replicates samples for each raw and boiled chicken livers, were prepared for the proximate analysis for the moisture, dry matter, ash, crude fat and crude protein content. All the analyses were carried out based on the certified procedures that outlined in Manual of Laboratory Techniques, University Putra Malaysia (2015), and additional referred to AOAC (1990). The crude fat proximate analysis was done with Soxhlet system whereas the


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crude protein proximate analysis was done with Kjeldahl’s method. There are total of twelve chicken livers for each of the raw and boiled chicken liver samples prepared for bacteria isolation and identification. All the data of the proximate analysis were analysed using IBM SPSS Statistic version 20. A probability of P < 0.05 will be accepted as statistically significant. RESULTS AND DISCUSSION The result had shown that all proximate analysis in both raw and boiled chicken livers with mean moisture content (raw: 76.29%, boiled: 72.11%), dry matter (raw: 23.71%, boiled: 27.89%), crude fat (raw: 13.15%, boiled: 12.57%) and crude protein (raw: 36.85%, boiled: 35.10%), were statistically difference, except for ash (raw: 1.48%, boiled: 1.45%) content percentage. The higher mean moisture, crude fat and crude protein in the raw chicken livers could be due to the heating process (boiling), which might cause the moisture, fat and protein loss in the chicken liver (Buff et al., 1997; Freeman et al., 2013). The variation of moisture and crude fat content of raw chicken liver as compared to some previous studies could be due to the different experiment setting (Abu-Salem et al., 2010) and age groups of the broiler chicken (Singh and Essary, 1974). The difference of crude fat and crude protein content in raw chicken liver also could be due to different sample processing method which causes the possibility of associated changes lipid oxidation and protein oxidation during frozen storage (Gray and Monahan, 1992; Uterera et al., 2013). The bacteria species isolated from raw chicken livers were Aeromonas species (100%), Streptococcus species (58.33%), Actinomyces species (50%), Proteus mirabilis (25%) and Salmonella species (16.67%). The presence of Salmonella species in the raw chicken livers was the main concern as most of the food-borne disease could be caused by Salmonellosis due to the consumption of raw foods. According to Arumugaswamy et al. (1995), 35% of liver among the raw diets in Malaysia, were contaminated with Salmonella species. REFERENCES: Abu-Salem, F. M., & Abou Arab, E. A. (2010). Chemical Properties, Microbiological Quality and Sensory

Evaluation of Chicken and Duck Liver Paste (foie gras). Grasas y Aceites, 61(2), 126-135. doi:10.3989/gya.074908

Buff, P. R., Carter, R. A., Bauer, J. E., & Kersey, J. H. (2014). Natural pet food: A review of natural diets and their impact on canine and feline physiology. Journal of Animal Science, 92(9), 3781-3791. doi:10.2527/jas.2014-7789

Freeman, L. M., Chandler, M. L., Hamper, B. A., & Weeth, L. P. (2013). Current knowledge about the risks and benefits of raw meat–based diets for dogs and cats. Journal of the American Veterinary Medical Association, 243(11), 1549-1558. doi:10.2460/javma.243.11.1549

Laflamme, D. P., Abood, S. K., Fascetti, A. J., Fleeman, L. M., Freeman, L. M., Michel, K. E., Willoughby, K. N. (2008). Pet feeding practices of dog and cat owners in the United States and Australia. Journal of the American Veterinary Medical Association, 232(5), 687-694. doi:10.2460/javma.232.5.687

Michel, K. E., Willoughby, K. N., Abood, S. K., Fascetti, A. J., Fleeman, L. M., Freeman, L. M., … Doren, J. R. (2008). Attitudes of pet owners toward pet foods and feeding management of cats and dogs. Journal of the American Veterinary Medical Association, 233(11), 1699-1703. doi:10.2460/javma.233.11.1699

Singh, S. P., & Essary, E. O. (1974). Factors Influencing Dressing Percentage and Tissue Composition of Broilers. Poultry Science, 53(6), 2143-2147. doi:10.3382/ps.0532143

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THE EFFECT OF DIFFERENT PROTEIN LEVELS ON THE PERFORMANCE OF MARDI KAMPUNG (VILLAGE) CHICKENS AT GROWER PHASE

N. SAMAT*, J. HAMED, A.A.I. AFFENDEE, S. RASOL, M.N. ISMAIL, R. ISMAIL AND

N. ISHAK Animal and Aqua Feed Program, Animal Science Research Center, Malaysian

Agricultural Research and Development Institute (MARDI), 43400, Serdang, Selangor. *Corresponding author: [email protected]

ABSTRACT

A study was conducted to investigate the effect of different crude protein (CP) levels on growth performance of MARDI kampung (village) chickens during grower phase. The experiment was conducted using completely randomized design with 4 treatments (17%, 19%, 21% and 23% CP), 4 replicates per treatment and with initial 24 birds per replicate. The level of dietary protein significantly affected the final body weight and body weight gain. The highest final body weight and body weight gain were achieved with 21% CP. The results also show that feed intake and FCR were not affected by different of dietary protein. It is concluded that dietary protein at 21 % with energy of 11 MJ/kg could be adopted to achieve high body weight for MARDI kampung chickens. Nevertheless, study is required to investigate whether or not different level of energy could improve further the performance of MARDI kampung chicken. Keywords: MARDI kampung chicken, growth performance, protein requirement, grower phase.

INTRODUCTION In Malaysia, most small holder and backyard farmers use commercial broilers diets to feed their kampung (village) chickens. The commercial broiler diets are formulated according to the standard nutrition requirements of fast growing chickens and most of these standards exceeded the needs of slow growing chickens, particularly kampung chickens (Liang et al., 1995). The use of existing commercial diets is not only contributing to the high cost of village chicken production, it also leads to nutrient wastage. Until now the diets formulated according to the requirement for kampung chicken breeds is not yet widely available. In the effort of producing cost effective formulations to suit kampung chicken breeds, nutritional requirements for commercial kampung chickens have been determined by several authors (Azahan et al., 2011, Termisal et al., 2012) but those reports focused on starter phase. While the nutritional requirements for MARDI kampung chickens, a newly breed developed via structured breeding program has not been reported elsewhere. Therefore, the objective of this study is to determine the effect of different level of dietary crude protein on the growth performance of MARDI kampung chicken at grower phase MATERIALS AND METHODS This experiment was conducted as a completely randomized design (CRD) arrangement of dietary treatments with 4 levels of crude protein namely 17, 19, 21 and 23 % crude protein (CP). Four corn-based diets were formulated to have different level of CP but isocaloric at 11 MJ/kg. Three hundred and eighty four 5-days old, mixed sex MARDI kampung chickens were weighed and randomly allocated to 4 treatments, each having 4 replicates with 24 birds per litter floor pen in a completely randomized design arrangement. The birds were fed the experimental diets in mash


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for 5 weeks starting from 4 to 9 weeks of age. Feed and water were available ad libitum at all times. Body weight and feed intake (FI) were measured weekly and body weight gains (BWG) and feed conversion ratio (FCR) were calculated. Data were analysed using general linear model (GLM) in Minitab 16, to assess the effects of different level of dietary CP with initial body weight at week 4 as co-variate. Significant differences among the treatments were identified at 95% confidence level by multiple comparison of Means Tukey’s Method. Test for normality was done by performing an Anderson-Darling Test. RESULTS AND DISCUSSION Table 1 shows that level of dietary protein significantly affected the final body weight and body weight gain. These parameters were gradually increased as dietary protein increased, peaked at 21% and reduced at 23% CP. This indicates 21% could be the optimum CP level with energy content of 11 MJ/kg to achieve highest bodyweight and weight gain. The results also show that different of dietary protein has no significant effect on feed intake and FCR, though there is numerically increased in feed intake and FCR as protein level increased. These findings contradicted those reported by Banerjee et al. (2013) where feeding Koekoeck chickens, a dual purpose free range breed, with increasing level of CP in isocaloric diets did not affected the overall body weight gain and final live weight of chickens. Table 1. Growth performance of MARDI kampung chickens fed on different level of dietary protein at 9 weeks old Level of dietary protein

Final body weights (g)

Body weight gain (g)

Feed intake (g) Feed conversion ratio

17% 1088.54c ± 7.86 702.67

c ± 10.90 2261.96 ± 77.40 3.22

± 0.09

19% 1162.50bc

± 24.10 782.29 bc

± 22.60 2179.60 ± 61.00 2.79 ± 0.08

21% 1205.57 a

± 7.58 822.76 a

± 9.43 2372.85 ± 85.60 2.88 ± 0.08

23% 1171.35 ab

± 8.22 788.54 ab

± 9.10 2481.15 ± 133 3.14 ± 0.13

Data were shown as mean ± standard deviation. Different superscript letters in each row indicate that there were significant differences (p<0.05), n=10

Based on this study, it is concluded that dietary protein at 21 % with energy of 11 MJ/kg could be adopted to achieve high body weight for MARDI kampung chickens. Nevertheless, study is required to investigate whether or not different level of energy could improve further the performance of MARDI kampung chicken. REFERENCES Banerjee, S., Melesse, A., Dotamo, E., Berihun, K. and Beyan, M. (2013) Effect of feeding different

dietary protein levels with iso-caloric ration on nutrients intake and growth performances of dual-purpose Koekoeck chicken breeds. Int. J. Appl. Poult. Res., 2(2): 27-32

Liang, J. B., Fierkens, B., Roch, J. J. and Engku Azahan, E. A. (1995) Efficiency of metabolisable energy utilisation by commercial broilers and indigenous chickens in Malaysia. MARDI Res. J. 23(1) (1995): 59–62

Engku Azahan, E.A., Azlina Azma, I.A. and Noraziah, M. (2011) Growth response of crossbred village (kampung) chickens to starter diets of differing energy contents. Mal. J. Anim. Sci. 14:51-55 (2011)

Tarmisal, A., E Engku Azahan, E.A., Azlina Azma, I.A. and Yusof Hamali, A. (2012). Utilization of a high protein starter feed by crossbred kampung chickens. Proc. 33

rd MASP Ann. Conf., 4-7 June 2012,

Langkawi, Malaysia 184-185.

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COMPARISON OF GROWTH PERFORMANCES OF BROILERS FED ON DIETS CONTAINING LOCAL AND IMPORTED GRAIN CORN

J. HAMED*, S. RASOL, A.H. ZAINUDIN, R. ISMAIL, N. ISHAK AND N. SAMAT Animal and Aqua Feed Program, Animal Science Research Center, Malaysian

Agricultural Research and Development Institute (MARDI), 43400, Serdang, Selangor. *Corresponding author: [email protected]

ABSTRACT

A study was conducted to determine the performance of the broiler chickens fed on local grain corn in comparison to imported grain corn. Two treatments, i.e. imported corn from Brazil and local corn (GWG 888) were tested as a completely randomised design (CRD) with 10 replications with 18 chickens per replicate. Weekly body weight and feed intake (FI) were measured and body weight gain (BWG) and feed conversion ratio (FCR) were calculated. Although the performance parameters between the treatments were not significantly different from each other, final body weight and BWG were shown to be higher in chickens fed on GWG 888, indicating feeding value of local corn has the tendency to be better than imported corn. In conclusion, the feeding value of local corn is comparable to those imported corn. Keywords: corn, feed intake, weight gain, feed conversion ratio.

INTRODUCTION Feed cost in poultry production has gone up substantially due to the increase in price of feed ingredients (Raghavan, 2009) and this situation has burdened the Malaysian poultry farmers. Grain corn is the main raw material in used in Malaysian poultry diets. Grain corn are imported amounting to 4 million metric tonnes worth RM 33.33 billion annually and most of them are from Argentina and Brazil. In the effort to reduce the deficit of this balance of trade, cultivation of grain corn in pilot scale was initiated in 2016. Several grain corn varieties were selected and cultivated at different locations all over Malaysia, mainly GWG 888, a local grain corn variety that was developed and produced by Malaysian corn breeder, Green World Genetic Sdn Bhd. Therefore, this study was conducted to determine the performance of the broiler chickens fed on local grain corn in comparison to imported grain corn. MATERIALS AND METHODS The work was undertaken in enclosed poultry house at Malaysian Agriculture Research and Development Institute (MARDI), Serdang, Selangor. Two treatments, which are imported corn from Brazil and local corn (GWG 888) were tested as a completely randomised design (CRD) with 10 replications with 18 chickens per replicate. The corn-based diets were formulated according to Cobb Nutrient Specification for starter and grower phases and were formulated to be isonitrogenous and isoenergetic. At one of age, male Cobb broiler chickens were weighed and randomly allocated to a total of 20 cages. The birds were fed the experimental diets in mash form for 42 days. Body weight and feed intake (FI) were measured and recorded weekly and body weight gains (BWG) and feed conversion ratio (FCR) were calculated. Meanwhile mortality data were monitored and recorded daily during the experiment period. Feed and clean water were given ad libitum at all time. The analysis of variance (ANOVA) was conducted using MINITAB software program version 14 and mean separation was determined using Turkey’s Test.


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RESULTS AND DISCUSSION The performance of broiler chickens fed on diets containing corn of different origins was shown in Table 1. Table 1: The performance of broiler chickens fed on diets containing corn of different origins

Treatment Imported corn (Brazil)

Local grain corn (GWG 888)

Initial body weight (kg)/bird 0.044 ± 0.001 0.044 ± 0.001 Final body weight (kg)/bird 2.490 ± 0.131 2.554 ± 0.107 Total body weight gain (kg)/bird 2.446 ± 0.129 2.510 ± 0.104 Total Feed intake (kg)/bird 4.638 ± 0.586 4.636 ± 0.512 Feed conversion ratio (FCR) 1.89 ± 0.192 1.85 ± 0.185

Data were shown as mean ± standard deviation. Different superscript letters in each row indicate that there were significant differences (p<0.05), n=10

The performance parameters between the treatments were not significantly different from each other. However, final bodyweight and body weight gain were shown to be higher in chickens fed on GWG 888, indicating feeding value of local corn has the tendency to be better than imported corn. In conclusion, the feeding value of local corn is comparable to those imported corn. REFERENCE Raghavan, V. (2009). Malaysia feed industry-issues and challenges. In Proceedings of 3rd International

Conferences on Animal Nutrition (ICAN) 2008: Enhancing Feed Utilization Through Technology (MARDI. Serdang), pp. 3-14.

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INCLUSION EFFECT OF SAGO FLOUR IN DIET ON GROWTH PERFORMANCE OF BROILER QUAILS (COTURNIX COTURNIX JAPONICA)

M.S. MUHAMMAD* AND M.N. HASSAN1

Animal and Aqua Feed Program, Animal Science Research Center, Malaysian Agricultural Research and Development Institute (MARDI), 43400, Serdang, Selangor 1Faculty of Agriculture and Food Science, Universiti Putra Malaysia, Bintulu Campus


ABSTRACT

Sago product has been identified as a potential carbohydrate source for poultry feed. The objective of this experiment is to determine the effect of different substitution level of sago flour in the diet on growth performance of quail. Ninety, mix sexes, day old quails were allocated into 3 treatment groups; Control (100% normal broiler diet, Cn), Treatment 1 (10% sago flour substitution - S10) and Treatment 2 (20% sago flour substitution -S20). Each treatment consists of 3 replications and housed in multiple battery raised floor cage system with 10 chicks per replication per cage. Substitution of sago flour in the diets significantly (P>0.05) affected the total feed intake but did not give any negative effect on the overall growth performance and carcass weight of quails. These observations clearly showed the potential of using sago as a food component in poultry feed particularly quail. In conclusion, sago flour could be in feeding poultry particularly quails. Further study should be done on the optimum inclusion level of sago flour in the feed to maximize the yield of the meat Keywords: quail, sago flour, growth performance.

INTRODUCTION The increase in feed prices and declining availability of grains for poultry has stimulated research into finding alternative feed resources to enhance cost-effective broiler productivity. Broiler quails (Coturnix coturnix japonica) are one type of poultry that present in various environments, from tropical rainforest and Europe temperate to Himalayan mountain area. In Malaysia, quail production is a very important business. There are 12 million quails has been produced in 130 quail farms in Malaysia and produce 350 million eggs per year. Feed grain like maize, rice, tapioca, wheat, sago and other grain and tuber are the main sources of carbohydrate for providing energy to maintain the performance of the quails (Abd-Aziz, 2002). The potential use of sago flour in feeding broiler quail has not be reported elsewhere. Therefore, the objective of this study is to determine the effect of different substitution level sago flour in the diet on the growth performance of quail. MATERIALS AND METHODS Ninety, mix sexes day-old broiler quail chicks were obtained from a commercial quail farm and were divided into 3 groups of treatment namely the control group (Cn1), treatment 1 (S10) and treatment 2 (S20). Each treatment consists of 3 replications in and housed in multiple battery raised floor cage system with 10 chicks per replication per cage and the treatments were arranged in a completely randomized design (CRD). Normal light bulb (60 watt) was put in place to provide heat for the chicks. In the control group, the quails were fed with 100% normal broiler starter feed and in S10 and S20 groups, the quails were fed with 10% sago flour mixed with 90% commercial feed and 20% sago flour was mixed with 80% normal feed, respectively. Every 5 days, the quails were individually weighed and the data were

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recorded. The weighing activity was held in the morning before the feed was offered. The feed residue was also collected and weighed. The feeding trough was cleaned before adding new batch of feed. After 45 days of experiment, the quails were slaughtered using halal method and the carcass was weighed without the feathers, skin and visceral organs. RESULTS AND DISCUSSION Substitution of sago flour in the diets significantly (P>0.05) affected the total feed intake. Feed intake was reduced as the substitution level increased. It was observed that quails in control group (Cn) have the highest feed intake (1151g), followed by 10% (1131g) and 20% substitution (1126g). Final average body weights of quails were also reduced as the level of substitution of sago flour increased from 327g per bird for 0 % substitution (control) to 320g per bird for 10% substitution and 302 g per bird for 20% substitution. The FCR at 45 days was 4.99, 5.05 and 5.36 for control, 10% and 20% substitution, respectively. However, the effect of sago flour substitution on final body weight and FCR was not significant. The average carcass weights were also found to be similar among the treatments i.e. 233g, 232g and 231g for carcasses from control, 10% and 20% groups, respectively. The present results indicate that the substitution of commercial broiler diet with sago flour did not give any negative effect on the overall growth performance and carcass weight of quails, except for the feed intake. The mortality rate was 0% and the quails showed no any nutrient deficiency symptom throughout the experiment. These observations clearly showed the potential of using sago as a food component in poultry feed particularly quail. In conclusion, sago flour could be in feeding poultry particularly quails. Further study should be done on the optimum inclusion level of sago flour in the feed to maximize the yield of the meat. REFERENCE Abd-Aziz, S. (2002). Sago starch and its utilisation. Journal of Bioscience and Bioengineering, 94(6),

526-529.

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OBSERVED AND EFFECTIVE NUMBER OF ALLELES ON COMMERCIAL KAMPUNG CHICKENS IN PENINSULAR MALAYSIA

I.A. AZLINA-AZMA* AND J.M. PANANDAM1

Animal and Aqua Feed Program, Animal Science Research Center, Malaysian Agricultural Research and Development Institute (MARDI), 43400, Serdang, Selangor

1Department of Animal Science, Faculty of Agriculture, Universiti Putra Malaysia 43400 UPM Serdang, Selangor Darul Ehsan


ABSTRACT

One hundred eighty birds from three commercial kampung chicken farms and 1 commercial broiler farm were analysed to measure their genetic relationship between each population using 27 microsatellite markers. Only 24 microsatellites are polymorphic with 2 to 9 alleles. The mean number of alleles per locus in KCA, KCB, KCC and CBR was 5, 5, 3 and 5 respectively. The effective number of alleles varied from 1.06 to 5.15 KCA, 1.06 to 5.88 KCB, 1.29 to 6.88 in KCC and 1.18 to 5.17 in CBR. Keywords: Kampung chicken, microsatellite markers.

INTRODUCTION Kampung chickens of today which are no longer purebred are resulted from uncontrolled breeding between the exotic breeds and the local kampung chicken, thus shows various phenotypic characteristics. At the end, no specific description that can be used to refer to these birds as a flock (Engku Azahan 1992). FAO (2004) recommended the conservation and protection of local AnGR to ensure the diversity of local/indigenous animals and the survival of the country’s legacy. In population genetics studies, molecular markers like microsatellite or simple sequence repeats (SSRs) or short tandem repeats (STRs) are used extensively worldwide. Kampung chickens have gone through random and indiscriminate mating in the earlier stages of their development followed by controlled and selected breeding in the later stages in the production of commercial kampung chicken. Using microsatellite markers, this study was carried out to evaluate observed and effective number of alleles on the commercial kampung chicken in Peninsular Malaysia. MATERIALS AND METHODS A total of 180 individual blood samples from three populations of kampung chicken (KCA = 50, KCB = 50 and KCC = 50) and one population of commercial broiler (CBr = 30) were used in this study. Genomic DNA was extracted from the blood using the Qiagen DNeasy Blood and Tissue Kit. 24 microsatellite loci (ADL0112, ADL0268, ADL0278, LEI0094, LEI0166, LEI0192, LEI0234, MCW0014, MCW0016, MCW0020, MCW0034, MCW0037, MCW0067, MCW0069, MCW0078, MCW0081, MCW0111, MCW0123, MCW0183, MCW0206, MCW0216, MCW0222, MCW0248 and MCW0295) were investigated in this study are those recommended by FAO (2004). PCR reactions were done in a final 25ml volume consist of 50 ng DNA template, 0.25 mM of each forward and reverse primers, 1 U of Taq polymerase, 0.25 mM of dNTP Mix, 1x buffer and 1.25 mM of MgCl2. PCR amplifications were performed as follows: initial denaturation at 95°C for 5 min, then followed by 40 cycles of denaturation step at 94°C for 50 s, annealing temperature (55 - 65°C) at 1 min and extension at

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72°C for 50 s and final extension 72°C for ten min. The PCR products were genotyped using 4% MetaPhor® Agarose (Lonza, USA) gel electrophoresis. The size of the bands representing the microsatellite markers was estimated using the gel documentation system (AlphaImager®) and the Alpha Ease software with reference to the 25 bp ladder marker. Using the POPGENE software version, data analysis on allele frequency, observed and expected mean heterozygosity (Nei, 1973). RESULTS AND DISCUSSION The mean number of alleles per locus in KCA, KCB, KCC and CBR was 5, 5, 3 and 5 respectively. The effective number of alleles varied from 1.18 at locus MCW0020 to 5.15 at locus LEI0192 in KCA population, 1.06 at locus MCW0020 to 5.88 at locus LEI0192 in population KCB, 1.29 at locus MCW0020 to 6.88 at locus LEI0192 and 1.18 at locus MCW0020 to 5.17 at locus LEI0192. According to Nevo (1978), number of alleles identified at each locus for each population can be used as an indicator of genetic variability. From the 27 microsatellites, only 24 microsatellites showed polymorphism with 2 to 9 alleles, with the highest number of alleles (9) observed in LEI0192 in KCC population. The total number of alleles in each population KCA, KCB, KCC and CBR are 110, 120, 125 and 110, respectively. Mukesh (2011) had used 15 microsatellite loci and reported a total of 173 alleles and with mean of 11.53 in their Red Jungle fowl in Northern India. While a study by Jian-Min (2010) on their Guangxi Three-yellow Chickens using 30 microsatellite loci had produced a total of 212 alleles with mean of 7.067. These two studies used different sets of microsatellite and populations. Number of mean observed alleles in the present study was lower than those reported by Mukesh (2011) and Jian-Min (2010) may be due to the low polymorphism that affected by long period of selection in the commercial kampung chicken production, which also happened to commercial layer population (Pirany et al. 2007). In conclusion, based on 27 microsatellite loci, three populations of commercial kampong chicken and a broiler population showed the genetic diversity and genetic relationship between these populations. REFERENCES Engku Azahan, E.A. (1992), Reproductive and productive performance of common varieties of

Malaysia kampung chicken. Proc. 6th

AAAP Animal Science Congress, 23-28th

Nov. 1992, Bangkok, Thailand. Vol 111:24.

FAO, (2004). Secondary guidelines for development of national farm animal genetic resources management plans for global management of cattle genetic resources using reference microsatellites global projects for the maintenance of domestic animal genetic diversity. Food and Agriculture Organization FAO, Rome, Italy (MoDAD). http://www.fao.org/dad-is/.

Mukesh, T., Kalasi, R.S., Mandhan, R.P. & Sathyakumar, S. (2011). Genetic diversity studies of Red Junglefowl across its distribution range in northern India. Asian J Biotechnol 3:293-301.

Nei, M., 1973 Analysis of gene diversity in subdivided populations. Proceedings of the National Academy of Sciences, USA 70: 3321-3323.

Nevo, E. (1978). Genetic variation in natural populations: patterns and theory. Theoretical population biology, 13(1), 121-177.

Pirany, N., Romanov, M. N., Ganpule, S. P., Devegowda, G., & Prasad, D.T. (2007). Microsatellite analysis of genetic diversity in Indian chicken populations. The Journal of Poultry Science, 44(1),19-28

Zou Jian-Min, Wei Feng-Ying, Shu Jing-Ting, Song Wei-Tao, Han Wei and Li Hui-Fang, (2010). Microsatellite DNA Typing for Assessment of Genetic Variability in Guangxi Three-yellow Chickens. Journal of Animal and Veterinary Advances, 9: 1565-1569.

http://www.fao.org/dad-is/

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DETECTION OF FUNGAL ORGANISMS IN PIGEONS AT SELECTED AREAS IN SELANGOR

A.R.N. NAZIHAH1,*, A. JALILA2, Z. ZUNITA2 , K. KRISHNAMMAH2 AND

R. RABIATULADAWIYAH2 1Department of Veterinary Clinical Studies; 2Department of Veterinary Pathology &

Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia


ABSTRACT The purpose of this study was to investigate the presence of fungal organisms in the pigeon droppings in selected areas in Hulu Langat & Kuala Langat, Selangor. The study demonstrated that pigeon droppings can be the platform for fungal growth and flourish. The fungal infection poses a significant public health threat as it can affect immunocompetent individuals and animals. Fifty samples of fresh pigeon droppings were obtained from exotic pigeons at the sampling area. Samples were transported to the Bacteriology laboratory in Faculty of Veterinary Medicine, Universiti Putra Malaysia and were immediately processed within 24 hours. Samples were cultures onto Sabouraud’s Dextrose agar and incubated at 37

o C for 15 days. Identification of cultures was determined using

Gram staining and API 20 C AUX identification test kits. Out of fifty samples, Candida species (88%), Trichosporon species (28%) and Saccharomyces species (6%) were isolated. In addition, mold fungi were also identified which include Aspergillus species (18%), Trichophyton species (6%), Mucor species (4%), Medurella species (1%) and Penicillium species (1%). This shows that the pigeons carry potential role as a reservoir for zoonotic yeasts and mold in the environment. Keywords: fungal, pigeon droppings, Selangor, isolation, identification.

INTRODUCTION Fungi are eukaryote that ingests food externally & absorb the nutrients through its cell wall (Carris et al., 2012). The fungi are able to cause disease due to their characteristics such as hyphae or spores. The invasive fungal infections cause morbidity and mortality in the immunocompromised host commonly through inhalation (Soltani et al., 2013). Pigeons may become the asymptomatic carries for fungal species. Furthermore, pigeon droppings that exist in the environment can provide the optimum condition for the organism to survive and flourish (Aiello L., 1967).

MATERIALS AND METHODS Fifty fecal samples were obtained from the exotic pigeons that kept in aviary located in Hulu Langat and Kuala Langat, Selangor. Samples were transported to the Bacteriology laboratory in Faculty of Veterinary Medicine, Universiti Putra Malaysia and were immediately processed within 24 hours. The isolation process was done where the samples were cultured in Sabauroud’s dextrose agar and incubated at 37o C for 15 days. Identification of cultures was determined using Gram staining and confirmation of yeast fungi was done using API 20 C AUX identification test kits. The mold fungi that grow on the agar plate were further identified using microscopic and macroscopic evaluation. In order to examine microscopically, lactophenol blue staining was performed. The results were represented by means of descriptive statistical analysis.


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RESULTS AND DISCUSSION Five genuses of yeast fungi were identified and the highest number to be found was Candida species followed by Trichosporon species. The Candida species are recognized as opportunistic pathogens which are normally found as micro-flora of humans and animals (Naz et al., 2017). Among 9 Candida species found, the Candida glabrata has the highest number but the species that has pathogenic importance would be the Candida albicans (Litmann et al., 1986) which found only in one sample. Instead found the Cryptococcus neoformans which most recognized pathogen isolated from pigeon droppings, Cryptococcus laurentii was isolated once. Even though there both have the same genus, the infection due to Cryptococcus laurentii is considered rare which had been reported in Molina et al. (2003). Other species were found categorized under opportunistic pathogen. Similarly, the mold fungi were categorized as opportunistic fungi and pathogenic fungi. The opportunistic fungi commonly encountered as contaminants and it is important to differentiate with the pathogenic fungi. The most frequently isolates fungi were followed by the Aspergillus species, Trichophyton species and Medurella species whereas for the opportunistic fungi; Penicillium species and Mucor species were isolated once in two samples. Aspergillus species has specific importance where it has the ability to produce aflatoxin that causes serious problem to animals as well as human (Abbas et al., 2017). For the other opportunistic fungus were found in this study, it shows that the environments have been contaminated (Soltani et al., 2013). The pathogenic fungi can be dangerous to the owner itself. However, as if the pigeons were kept in a confined area, the spread of the pathogenic fungi could be minimized. The initial source of the fungi could not be identified as the pigeons were kept in confined area in a long period of time. These results demonstrate that pigeon droppings are an important and environmentally source of yeast and mold fungi (Soltani et al., 2013) in two districts in Selangor. As prevention and control for the pathogenic fungi, the waste management should be cleaned frequently to reduce the number of opportunistic fungi in the birdhouse. In the same time, the owner should wear protected mask and gloves while handling the feces are essential to reduce transmission of the pathogen to humans. REFERENCES Ajello L. (1967). Comparative ecology of respiratory mycotic disease agents. Bacteriological

reviews,31(1),6. Carris, L.M., Little, C.R. and Stiles, C.M. (2012) Introduction to Fungi. The Plant Health Instructor. Molina-Leyva, A., Ruiz-Carrascosa, J. C., Leyva-Garcia, A., & Husein-Elahmed, H. (2013).

Cutaneous Cryptococcus laurentii infection in an immunocompetent child. International Journal of Infectious Diseases, 17(12), e1232-e1233

Soltani, M., Bayat, M., Hashemi, S. J., Zia, M., & Pestechian, N. (2013). Isolation of Cryptococcus neoformans and other opportunistic fungi from pigeon droppings. Journal of research in medical sciences: the official journal of Isfahan University of Medical Sciences, 18(1), 56.

Weber, W. (1979).Pigeon associated people diseases. In Bird Control Seminars Proceedings, 21. Littman, M. L., & Borok, R. (1968). Relation of the pigeon to cryptococcosis: natural carrier state,

heat resistance and survival of Cryptococcus neoformans. Mycopathologia et mycologia applicata, 35(3-4), 329-345.

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PRELIMINARY STUDY ON EFFECT OF FEEDING MILK EXTRACTED COCONUT MEAT ON EGG PRODUCTION OF KHAKI CAMPBELL DUCKS

M.N. AZLIAN*, B.M. NORHAFIFI, A.P.M. SYAHMI, A.A. HAKIM AND N. SAMAT

Animal Science Research Centre, Malaysian Agriculture Research and Development Institute, 43400, Serdang, Selangor, Malaysia *Corresponding author: [email protected]

ABSTRACT

The study carried out to determine the effect of feeding milk-extracted coconut meat on egg production of Khaki Campbell ducks in their second year of production. Ninetysix (96) ducks were randomly distributed to 6 pens and the pens were randomly assigned using a completely randomized design (CRD) to two experimental diets; Control (100% basal diet) and Treatment (basal diet + 10% coconut meat) with 3 replications. Daily egg production and hen-day production for the period of experiment were not influenced by feeding ducks with 10% coconut meat (p>0.05). In conclusion, milk-extracted coconut meat has a potential to be a substitute feed ingredient in feeding old laying duck to lower cost of egg production. Further study is reckoned to determine the effect of milk-extracted coconut meat on egg quality Keywords: coconut meat, Khaki Campbell ducks, egg production.

INTRODUCTION Global egg production has increased dramatically over the last 20 years with Asia is taking the lead (FAO, 2014). Egg consumption in Malaysia has increased over past 5 years from 7,887 million to 9,922 million eggs in 2009 and 2014, respectively (DOA, 2016). Feed accounts for about 70% of the total cost of production of livestock. Backyard duck farming is having lower production level and less profitable due to shortage of feed and high cost involved in the procurement and transport of raw materials or readymade feed materials. Thus, it is a critical need for an alternative feed material that could be used alongside the existing commercial feed thereby to reduce the feed cost. In order to maintain productivity at a lower feed cost, cheaper and readily available alternative feedstuffs should be studied for their potential in livestock diets. In Malaysia, about 151,000 ha of land was being used for coconut plantation in year 2001, estimated that 5,280 kg of dry coconut meat become available per hectare per year. The protein level of coconut meat typically ranges from 20 to 26 % and contain crude fibre between 10 and 16 %. Gross energy in coconut husk is greater than in corn i.e. 18 MJ/kg but the metabolisable energy is less than in corn due to high fibre content (Stein et al., 2015). The objective of this study is to determine the effect of feeding milk-extracted coconut meat on egg production of Khaki Campbell ducks in their second year of production.

MATERIALS AND METHODS The experiment was conducted at Organic Farm of MARDI Serdang. A total of 96 Khaki Campbell ducks with mean body weight of 1281.70 ± 146.52g were randomly allocated into six different pens with 15 to 16 ducks per floor pen. The pens were randomly assigned using a completely randomized design (CRD) to two experimental diets; Control (100% basal diet) and Treatment (basal diet + 10% coconut meat) with 3 replications. Grated mature coconut meat used in this study was the waste from coconut milk extraction and was obtained from local wet market. The coconut meat

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was dried at 60oC overnight after being separated from hard shell, husks and other impurities. The experiment was carried out for 2 weeks and the ducks were fed ad libitum. Drinking water was available all the time. Feed intake per pen and number of egg produced were recorded and the hen-day egg production were calculated. All data were subjected to one-way ANOVA to compare the differences between 2 treatments. Mean separations were determined using Turkey’s Test. The statistical analysis was conducted using MINITAB software program version 17. RESULTS AND DISCUSSION Table1. Egg production and percent hen-day production of Khaki Campbell ducks fed with coconut husk

Treatment Daily Egg Production (no of eggs) % hen-day production

Control 9.05 ± 0.42 58.95 ± 1.56 10% coconut husk 8.67 ± 2.39 54.20 ± 15.00 Data were shown as mean ± standard error. Different superscript letters in each row indicate that there were significant differences (p<0.05), n=3

Daily egg production and hen-day production for the period of experiment were not influenced by feeding ducks with 10% coconut meat (p>0.05). Ducks fed with 10% coconut meat have similar number of egg laid daily and hen-day production percentage as those fed on diet without coconut meat, although those values were numerically lower. This indicates that coconut meat could be fed to laying ducks without adversely affecting the egg production. These findings was in line with Cocjin (1991), who observed that egg production of younger Mallard ducks did not vary significantly when 25% and 50% of fresh coconut meat (milk was not extracted) were included in the diets in comparison to those without coconut meat. In conclusion, milk-extracted coconut meat has a potential to be a substitute feed ingredient in feeding old laying duck to lower cost of egg production. Further study is reckoned to determine the effect of milk-extracted coconut meat on egg quality. REFERENCES Cocjin, B.B. (1991). Fresh coconut meat in poultry ration. Asian-Australasian Journal of Animal

Sciences 1991;4(2): 187-193. DOA 2016. Maklumat telur ayam dan itik 2009-2014

http://www.data.gov.my/data/dataset/2162612e.

FAO (2014). Assessed at www.fao.org/publications on 12/10/2015.

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ACKNOWLEDGEMENTS

The Chairman and the Organizing Committee wish to express their utmost gratitude to the following individuals, companies and organizations who have contributed in

one way or another in ensuring the success of this conference.

Thank you.

World’s Poultry Science Association (Malaysia Branch)

World Veterinary Poultry Association (Malaysia Branch)

Faculty of Veterinary Medicine, Universiti Putra Malaysia

Department of Veterinary Services, Malaysia

United Business Media (M) Sdn Bhd

Livestock Asia Expo & Forum 2018

Cevac IBird

AGCO GSI (Malaysia) Sdn Bhd

TLC Veterinary Services

Distinguished session chairman, speakers, poster presenters and participants

All relevant parties and individuals who have contributed to the success of this

conference

We look forward to meeting you again.

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enhancing poultry health and production for sustainable ... · p.x. lim and n.m.a.n.m. faiz 73...

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