I Gusti Ngurah Kade Mahardika
Laboratorium Biomedik Fakultas Kedokteran Hewan Universitas Udayana, Denpasar

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PERBANDINGAN SEKUENS KONSENSUS GEN HEMAGLUTININ VIRUS AVIAN INFLUENZA SUBTIPE H5N1 ASAL UNGGAS DI INDONESIA DENGAN SUBTIPE H5N2 DAN H5N9 Kade Mahardika, I Gusti Ngurah; Suartha, I Nyoman; Suardana, Ida Bagus Kade; Yuniati Kencana, I Gusti Ayu; Teguh Wibawan, I Wayan
Jurnal Veteriner Vol 10 No 1 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Consensus sequence of hemagglutinin (HA) gene of avian influenza viruses of H5N1 subtype isolatedfrom fowl in Indonesia ? hereafter named as H5N1_Indonesia ? is compared with that of H5N2 and H5N9viruses. Sequence information were downloaded from the public database GeneBank. The genetic distancesand nucleotide as well as its deduced amino-acid sequence alignment were analysed. At nucleotide level,genetic distances of HA between H5N1_Indonesia to H5N2 and H5N9 are 16.2% and 9.6%, respectively.At amino-acid level, the distances are 9.7% and 6.8%. The genetic distances of HA1 fragments are 19.0%(H5N1_Indonesia ? H5N2) and 10.9% (H5N1_Indonesia ? H5N9). At amino-acid, level the genetic distancesof HA1 are 13.1% (H5N1_Indonesia-H5N2) and 8.8% (H5N1_Indonesia ? H5N9). All three subtypes havedifferent glycosilation motive and variation of amino-acid sequence of four antigenic sites. The consequenceof those facts is discussed.
RESPON IMUN ITIK BALI TERHADAP BERBAGAI DOSIS VAKSIN AVIAN INFLUENZA H5N1 Kade Suardana, Ida Bagus; Ritha Krisna Dewi, Ni Made; Kade Mahardika, I Gusti Ngurah
Jurnal Veteriner Vol 10 No 3 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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A study was carried out to investigate the immune response of Bali ducks against various doses ofAvian Influenza H5N1 vaccine. The study was carried out using a complete Random-Split in Time researchdesign as many as 40 of Bali ducks of 3 months age were kept separately in 4 groups. The ducks werevaccinated twice in two week interval with AI H5N1 vaccine of 0 (as negative control), 1/2, 1, and 2 doses.Sera were collected one day before first vaccination, then every week until three weeks after the secondvaccination. All sera were tested by hemaglutination inhibition (HI) test. The result shows that antibodylevel with double dose was significantly higher than single dose, half dose, and negative control (P<0.01).However antibody level in ducks vaccinated with single and half dose did not show any significant difference(P > 0.05).
PHYLOGENETIC AND ANTIGENIC STRUCTURE OF AVIAN INFLUENZA VIRUS OF H5N1 SUBTYPE ISOLATED FROM WATERFOWLS Susanti, R; Damajanti Soejoedono, Retno; Kade Mahardika, I Gusti Ngurah; Wibawan, I Wayan Teguh; Thenawidjaja Suhartono, Maggy
Jurnal Veteriner Vol 9 No 3 (2008)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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A study was carried (1) to analyze the phylogenetic relationship of fragment hemaglutinin (HA) geneof avian influenza viruses (AIV) subtype H5N1 isolated from apparently healthy backyard waterfowls inWest Java with representative of animal and human isolates from Indonesia and some countries in Asia;(2) to find out cross-reactivity of those viruses with a standard Indonesian strain. Nucleotide sequences ofHA gene of AIV H5N1 from backyard waterfowls along with other H5N1 isolates of Indonesian and Asianorigin were aligned using with ClustalW of MEGA 3.1 program. Estimation of genetic distance and theconstruction phylogenetic tree were conducted by Neighbor Joining method and calculation of distancematrix using Kimura 2-parameter. Antigenic analysis was conducted using hemagglutination inhibition(HI) test. Result of phylogenetic analysis indicated that all viruses from backyard waterfowls form threedistinct sublineages. One lineage was located in Indonesia cluster and two lineages in Asia cluster. In thephylogenetic analysis, it was concluded that multiple introductions of AIV H5N1 to Indonesia have occurred.Six AI H5N1 viruses from backyard waterfowls (IPB1-RS to IPB6-RS) appeared to be different ancestorsthose isolated previously in Indonesia. Cross-antigenic analysis showed that nine viruses isolates used inthis study were antigenically different to Legok 2003 chicken strain of AIV H5N1. The HI titer of anti-Legok 2003 antibody with all newly isolated viruses is up to 6 log lower then the HI titer using homologstrain.
AMINO ACID SEQUENCE MOTIVE OF OSELTAMIVIR BINDING POCKET IN NEURAMINIDASE PROTEIN OF AVIAN INFLUENZA (H5N1) VIRUS FROM HUMAN AND ANIMAL IN INDONESIA Kade Mahardika, I Gusti Ngurah; Sukada, I Made; Suma Antara, Made; Suartini, I Gusti Ayu
Jurnal Veteriner Vol 9 No 4 (2008)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Former finding that avian influenza (AI) virus of H5N1 subtype from Indonesia shows reduced sensitivity against oseltamivir is critically reviewed trough molecular observation of the amino-acid sequence motive of neuraminidase protein (NA) of all H5N1 virus from human and animal in Indonesia available in GeneBank. Amino acid sequence of oseltamivir binding pocket of NA protein on all Indonesian viruses is typical for sensitive virus with a concerved motive of H274, E276, R292 dan N294. Resistance issue could not be explained based on available data.
PERANAN PEDAGANG UNGGAS DALAM PENYEBARAN VIRUS AVIAN INFLUENZA Suartha, I Nyoman; Suma Antara, I Made; Saka Wiryana, I Kadek; Sukada, I Made; Wirata, I Wayan; Ritha Krisna Dewi, Ni Made; Kade Mahardika, I Gusti Ngurah
Jurnal Veteriner Vol 11 No 4 (2010)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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A questionnaire surveillence have been carried out in three different traditional markets (ie. Beringkitin badung district, Kumbasari in Denpasar, Kediri in Tabanan district) in order to understand the role ofpaultry traders behavior in transmitting of avian influenza virus. Of 150 quationares collected most oftraders (66.7%) kept the animals for 1-3 days before it was marketed. Traders bin Beringkit and Kediri(76.3%) used to mix different species of birds in their cages, whereas none of the traders from Kumbasaridoing that. When hygienec and sanitation aspects were considered (ie. Washing and desinfectan sprayingfor cages) it was found that the behavior of traders varied markedly between the 3 different market. Inconclusion the traders awareness to especially bird flue infection and implementation of biosecurity isvery low.
POLA DISTRIBUSI UNGGAS DARI PASAR TRADISONAL BERPERAN DALAM PENYEBARAN VIRUS FLU BURUNG SUMA ANTARA, I MADE; SUARTHA, I NYOMAN; SAKA WIRYANA, I KADEK; SUKADA, I MADE; WIRATA, I WAYAN; DIBYA PRASETYA, I GUSTI NGURAH; RITHA KRISNA DEWI, NI MADE; KOMALA SARI, TRI; KADE MAHARDIKA, I GUSTI NGURAH
Jurnal Veteriner Vol 10 No 2 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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A study has been carried out to map the distribution pattern of poultry from traditional market toreduce the transmission risk of avian influenza virus. The data were collected from threes markets wherepoultry are sold, namely in Bringkit of Badung Regency, Kumbasari of Denpasar City, and Kediri ofTabanan Regency. Data collections was based on interviews using questionnaire. Poultry from all marketsare distributed throughout Bali. Poultry are traded mainly for religious ceremony and immediatelyslaughtered as it arrives at the consumer?s house. The distribution pattern of poultry seems to play asignificant role in the disseminations of avian influenza virus. The right implementation of biosecurity intraditional markets is highly recommended to curb the risk.
AMINO-TERMINUS OF POLYMERASE BASIC-2 OF AVIAN INFLUENZA VIRUS OF H5N1 SUBTYPE ISOLATED FROM VARIOUS ANIMAL SPECIES IN INDONESIA Yuniati Kencana, Gusti Ayu; Asmara, Widya; Rangga Tabbu, Charles; Kade Mahardika, I Gusti Ngurah
Jurnal Veteriner Vol 9 No 3 (2008)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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The information on pathogenicity and adaptation factors of avian influenza virus (AIV) in mammalsis very inportant in an effort to reduce the risk of avian influenza (AI) pandemic in the future. Polymerasegene complex appears to be the major factors for adaptation of AIV to certain animal species. A preliminarystudy on role of non-coding region (NCR) and amino-terminus of polymerase-basic 2 (PB2) is presented.Purified viral RNA of AIV isolated from chicken, duck, pig, and quail of Bali and Yogyakarta was reversetranscribed into cDNA and amplified using reverse transcriptase-polymerase chain reaction (RT-PCR)using PB-2 universal forward primer and specifically designed backward primer. The result showed thatall AIV?s H5N1 isolated from chicken, duck, quail, and pig, posed PB2 amino-terminus typical for IndonesianAIV H5N1. However, polymorphic amino acids of the protein fragment did not show any species specificmotive, with the exception of the pig isolate Sw/Tabanan/2006 which had specific substitution of D16E,H17Q, M40I, and H124Y.
PENEGUHAN DIAGNOSIS PENYAKIT NEWCASTLE DISEASE LAPANG PADA AYAM BURAS DI BALI MENGGUNAKAN TEKNIK RT-PCR Kencana, Gusti Ayu Yuniati; Kardena, I Made; Kade Mahardika, I Gusti Ngurah
Jurnal Kedokteran Hewan Vol 6, No 1 (2012): J. Ked. Hewan
Publisher : Syiah Kuala University

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Abstract

Penelitian ini bertujuan mendiagnosis kasus newcastle disease (ND) lapangan pada ayam bukan ras yang bersifat akut melalui hasil pemeriksaan di Laboratorium Patologi dan Laboratorium Biomedik Fakultas Kedokteran Hewan Universitas Udayana tahun 2008-2009. Sebanyak sepuluh ekor sampel ayam buras telah diperiksa. Gejala klinis yang teramati meliputi: anoreksia, lesu, bersin, batuk, dan diare putih kehijauan dengan diagnosis sementara sebagai penyakit ND yang bersifat akut. Sampel diambil dari organ yang mengalami perubahan patognomonis seperti pada proventrikulus, ventrikulus, seka tonsil, paru-paru dan otak. Perbanyakan virus menggunakan telur ayam bertunas (TAB) umur 10 hari melalui ruang alantois dan diinkubasikan pada inkubator telur suhu 37° C selama 3 hari. Koleksi cairan alantois dilakukan pada hari ke-3, selanjutnya diidentifikasi dengan uji serologi hemaglutinasi (Haemaglutination-Inhibition Test/HA/HI) dengan teknik mikrotiter baku dan dikonfirmasi dengan uji reverse transcriptase-polymerase chain reaction (RT-PCR) menggunakan primer FNDIFP (5’-CCCCGTTGGAGGCATAC-3’) dan FNDIBP (5’-TGTTGGCAGCATTTTGATTG-3’). Hasil penelitian menunjukkan bahwa semua sampel kasus ayam bukan ras yang diperiksa positif terinfeksi oleh virus penyakit ND akut. Kajian ini mengindikasikan bahwa penyakit ND di Bali masih bersifat endemis.
ANALISIS SEKUENS D-LOOP DNA MITOKONDRIA SAPI BALI DAN BANTENG DIBANDINGKAN DENGAN BANGSA SAPI LAIN DI DUNIA DWINA WISESA, ANAK AGUNG NGURAH GEDE; KADE MAHARDIKA, I GUSTI NGURAH; OKA PEMAYUN, TJOK GEDE
Indonesia Medicus Veterinus Vol 1 (2) 2012
Publisher : Faculty of Veterinary Medicine, Udayana University

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The result of this study indicates the D-loop of mtDNA of bali cattle is homologous with banteng. Two haplotypes can be identified. The sequence is different from that of Bos taurus, Bos indicus, Bos javanicus, and Bos gaurus. Further research needs to be done with a representative sample for the entire population in Bali by targeting more than one locus. In addition, the effective population size of bali cattle needs to be known to improve the genetic quality of bali cattle.
PRODUKSI IGY ANTIVIRUS AVIAN INFLUENZA H5N1 DAN PROSPEK PEMANFAATANNYA DALAM PENGEBALAN PASIF Teguh Wibawan, I Wayan; Murtini, Sri; Damajanti Soejoedono, Retno; Kade Mahardika, I Gusti Ngurah
Jurnal Veteriner Vol 10 No 3 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Immunoglobulin Y (IgY) in yolk has been shown in several studies to prevent both bacterial and viralinfections. This research was conducted to find evidence that IgY specific against avian influenza virus(AIV) of H5N1 subtype can be produced in a large quantity in egg yolk. Laying hens were vaccinated withAI killed-vaccine (IPB-Shigeta). The IgY was purified using affinity chromatograpy technique, and anti-H5activity was measured using a standard haemaglutination inhibition (HI) and agar gel immunodifusion.The concentration of IgY was calculated, and the protein pattern was detected using polyacrilamid gel(AGID) electrophoresis (PAGE). Anti H5 antibody as high as 27 ? 29 HI units was detected and produce aspecific line of precipitation in AGID. The concentration of IgY was 7.89 mg/ml. Purified specific IgY consistof 6 main protein bands with molecular weights ranging from 35 to 225 kD. These proteins were sensitiveto heat treatment (75oC for 30 minutes), to acid condition (pH2) as well as the pepsin and trypsin. Theseresults indicated the possibility of using specific IgY for passive immunisation to prevent AIV infection oras immunotherapeutic applications for AI treatment in humans.