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All Journal HAYATI Journal of Biosciences ACTA VETERINARIA INDONESIANA Jurnal Sain Veteriner Jurnal Veteriner Jurnal Ilmu-Ilmu Peternakan (Indonesian Journal of Animal Science) Indonesian Journal of Biotechnology Widyariset Widyariset Jurnal Kedokteran Hewan Journal of Veterinary and Animal Sciences

Michael Haryadi Wibowo

Departemen Mikrobiologi, Fakultas Kedokteran Hewan, Universitas Gadjah Mada

Author-ID : 277276

Aerospace Engineering Agriculture, Biological Sciences & Forestry Astronomy Automotive Engineering Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Civil Engineering, Building, Construction & Architecture Computer Science & IT Control & Systems Engineering Decision Sciences, Operations Research & Management Earth & Planetary Sciences Electrical & Electronics Engineering Energy Engineering Immunology & microbiology Industrial & Manufacturing Engineering Library & Information Science Materials Science & Nanotechnology Mechanical Engineering Medicine & Pharmacology Neuroscience Physics Veterinary Other

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The Development of Pathogenicity of Avian Influenza Virus Isolated from Indonesia Wibowo, Michael Haryadi; Srihanto, Agus Eko; Putri, Khrisdiana; Asmara, Widya; Tabbu, Charles Rangga
Indonesian Journal of Biotechnology Vol 18, No 2 (2013)
Publisher : Universitas Gadjah Mada

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Highly pathogenic avian infl uenza outbreak in Indonesia has been reported in various poultry due toH5N1 subtype. The presence of multiple basic amino acids within the cleavage site of HA glycoprotein hasbeen identifi ed to be associated with the pathogenicity of avian infl uenza virus. The study was retrospectivestudy which was designed to characterize the cleavage site and fusion site region of haemagglutinin gene ofAIV isolated from various poultry in 2003 to 2013. Isolation, Identifi cation and propagation were carried outto collect viral stock. For virus detection, reverse transcriptase PCR (RT-PCR) method on H5 and N1 genefragment was performed. All of RT-PCR HA gene positive products were sequenced for further nucleotideanalysis and to determine the nucleotide composition at the targeted fragment. The results are all AIV isolateswere identifi ed as H5N1 subtype. The sequence analyses revealed some motives of basic amino acid motivethat were classifi ed as highly pathogenic avian infl uenza virus. Further analyses on fusion domain of all AIVisolated during the period 2003 to 2013 showed conserved amino acid.Keywords: avian infl uenza, haemagglutinin, cleavage site, basic amino acid, fusion site

Analisis Fragmen Gen VP-2 Virus Infectious Bursal Diseases yang Diisolasi dari Peternakan Ayam Komersial Michael Haryadi Wibowo; Dito Anggoro; Sarwo Edy Wibowo; Purnama Edy Santosa; Surya Amanu; Widya Asmara
Acta VETERINARIA Indonesiana Vol. 5 No. 1 (2017): Januari 2017
Publisher : IPB University

Infectious Bursal Disease (IBD) adalah penyakit virus yang bersifat akut dan infeksius serta menyerang pada unggas muda yang berumur kurang dari 4 bulan. Sejauh ini data molekuler virus IBD isolat Indonesia sangat minim, oleh karena itu penelitian ini bertujuan untuk mengidentifikasi dan mengarakterisasi gen VP-2 virus IBD yang telah ada di Indonesia. Sampel penelitian diperoleh dari kasus terdiagnosa IBD yang terjadi di peternakan ayam komersial broiler dan layer. Sampel Bursa dipersiapkan untuk dilakukan isolasi menggunakan telur ayam berembrio SPF. Membran korioalantois dipanen dan dilakukan identifikasi dengan metode RT-PCR dengan gen target VP-2. Hasil amplifikasi selanjutnya dilakukan pengurutan DNA. Data nukleotida hasil pengurutan DNA dianalisis dengan program MEGA 6, meliputi pesejajaran, prediksi asam amino, dan konstruksi pohon kekerabatan antara virus yang diteliti dengan beberapa virus yang telah dipublikasi di bank gen terutama virus IBD yang bersirkulasi di Indonesia. Hasil penelitian ini diperoleh data bahwa ayam yang terdiagnosis penyakit IBD dapat ditentukan penyebabnya sebagai virus IBD. Hasil analisis urutan penanda patogenisitas molekuler menunjukkan virus yang virulen. Analisis pohon kekerabatan 2 isolat IBD SR/Lay-WNO-DIY dan IBD Potrow/Lay-SLM-DIY termasuk dalam kelompok virus IBD tipe klasik, sedangkan lima virus lainnya, yaitu IBD Yanti/Lay-SLM-DIY; IBD Lampung/Bro/IL; IBD Fung/Lay-SLM-DIY-BF1, IBD Fung/Lay-SLM-DIY-BF2, dan IBD Fung/Lay-SLM-DIY-BF3 termasuk dalam kelompok vvIBD strain Indonesia.

Penentuan Patogenesitas Molekuler Virus Newcastle Disease yang Diisolasi dari Ayam Komersial Tahun 2013-2016 Sarwo Edy Wibowo; Michael Haryadi Wibowo; Bambang Sutrisno
Acta VETERINARIA Indonesiana Vol. 5 No. 2 (2017): Juli 2017
Publisher : IPB University

Newcastle Disease (ND) atau yang dikenal dengan “Tetelo” masih menjadi masalah di peternakan unggas komersil, meskipun telah dilakukan vaksinasi ND secara rutin, namun wabah ND masih tetap terjadi. Penelitian ini bertujuan untuk mengetahui patogenesitas molekuler dan genotipe virus ND berdasarkan analisis sekuen fragmen gen F, serta melihat hubungan kekerabatan antara isolat dalam penelitian dengan isolat ND di Indonesia sebelumnya serta strain vaksin pada peternakan ayam yang menerapkan vaksinasi ND secara berkala. Penelitian ini menggunakan primer yang didesain dengan konsensus fragmen gen F dari GenBank dan desain dengan aplikasi amplifX pada posisi 91-800 nt dengan panjang 710bp. Urutan basa pada primer kemudian di cek dengan BLAST primer dan di uji spesifisitas dengan beberapa virus penyakit unggas yaitu vaksin infectious bronchitis (IB) 120, virus vaksin infectious laryngotracheitis (ILT), virus vaksin avian influenza (AI), dan virus vaksin infectious bursal disease (IBD). Delapan sampel paru diperoleh dari delapan peternakan ayam komersial di Yogyakarta, Semarang, Jakarta, Magelang dan Muntilan. Tiga sampel diisolasi pada telur ayam berembrio dan diidentifikasi menggunakan uji HA dan HI. Lima sampel lain dilakukan ekstraksi secara langsung dari gerusan organ paru. Sampel di identifikasi dengan metode reverse transciptase-polymerase chain reaction (RT-PCR) untuk mendeteksi gen F menggunakan primer forward 5’ TCT CTT GAT GGC AGG CCT CTT G ‘3 dan reverse 5’ CCG CTA CCG ATT AAT GAG CTG AGT’3 dengan panjang produk 710 bp. Hasil penelitian menunjukkan bahwa seluruh sampel positif virus ND. Analisis hasil sekuen dengan menggunakan perangkat lunak MEGA v.7 didapatkan susunan asam amino penyusun cleavage site 112RRQKR↓F117 dan 112RRRKR↓F117yang menunjukkan bahwa virus ND tersebut digolongkan strain velogenik. Berdasarkan analisis pohon filogenetik isolat ND-Layer/GK-SR1/2013, ND-Lay/Pullet-80/ 27/16 (N), ND-Bro/Lingga 2L/24/2 (N), dan ND-Lay/Smg-P/2015 merupakan genotip VIIi, sedangkan 3 isolat ND yaitu ND-Bro/Yog-P/2015, JKT/P1/2016 (Jakarta), dan JKT/P2/2016 (Jakarta) merupakan ND genotip VIIh. Jarak genetik isolat yang diteliti dengan virus ND Indonesia yang pernah dilaporkan sebelumnya pada fragmen gen F posisi 91-798 berkisar 0,4 – 9,6 % dengan tingkat homologi mencapai 90,4 – 99,5%.

The Development of Pathogenicity of Avian Influenza Virus Isolated from Indonesia Michael Haryadi Wibowo; Agus Eko Srihanto; Khrisdiana Putri; Widya Asmara; Charles Rangga Tabbu
Indonesian Journal of Biotechnology Vol 18, No 2 (2013)
Publisher : Universitas Gadjah Mada

Highly pathogenic avian infl uenza outbreak in Indonesia has been reported in various poultry due toH5N1 subtype. The presence of multiple basic amino acids within the cleavage site of HA glycoprotein hasbeen identifi ed to be associated with the pathogenicity of avian infl uenza virus. The study was retrospectivestudy which was designed to characterize the cleavage site and fusion site region of haemagglutinin gene ofAIV isolated from various poultry in 2003 to 2013. Isolation, Identifi cation and propagation were carried outto collect viral stock. For virus detection, reverse transcriptase PCR (RT-PCR) method on H5 and N1 genefragment was performed. All of RT-PCR HA gene positive products were sequenced for further nucleotideanalysis and to determine the nucleotide composition at the targeted fragment. The results are all AIV isolateswere identifi ed as H5N1 subtype. The sequence analyses revealed some motives of basic amino acid motivethat were classifi ed as highly pathogenic avian infl uenza virus. Further analyses on fusion domain of all AIVisolated during the period 2003 to 2013 showed conserved amino acid. Keywords: avian infl uenza, haemagglutinin, cleavage site, basic amino acid, fusion site

Evaluation of rapid detection kit against avian influenza A virus and H5 subtype for field Sample Michael Haryadi Wibowo; Tri Untari; Sidna Artanto; Krisdiana Putri; Surya Amanu; Widya Asmara
Indonesian Journal of Biotechnology Vol 21, No 1 (2016)
Publisher : Universitas Gadjah Mada

Avian influenza virus is poultry viral disease, which causes high economic losses. Various efforts have been made to control the disease. One effort is required screening fast and precise diagnostic test. This study was aimed to determine the potential of rapid test kit of AIV/H5 Anigen Rapid Test for the detection of AI virus types A and subtype H5 in the field. Some tests were carried out, e.g.: the potential test, cross-reaction test, sensitivity and specificity test. Potency test was done to evaluate potential of detection limits of the kit, by having the test of serial dilution of AI virus positive control. Cross-reaction test was done to detect antigens other than AI virus H5N1, e.g.: IB virus of Massachuset strain, IBV strain 4-91, Newcastle Disease virus, and Escherichia coli. Sensitivity and specificity test were applied to the filed samples which clinically and laboratory were confirmed as H5N1 positive. To confirm the result of rapid test was then being done by reverse transcriptase polymerase chain reaction. Based on these results it can be concluded that, Anigen Kit AIV/H5 Ag Rapid Test can detect antigen-containing samples having AI virus HA titer up to 26of type A virus, and up to 25 for subtype H5 virus. Anigen Kit AIV/H5 Ag Rapid Test showed no cross-reactions with Infectious Bronchitis virus, Newcastle Disease virus, and Escherichia coli. Anigen A Rapid Test Kit AIV Ag showed a sensitivity of 50% and specificity of 100%, while Anigen Ag Rapid Test Kit AIV/H5 showed a sensitivity of 25% and specificity is 100%.

The establishment of PCR amplification, cloning, and sequencing of bovine herpesvirus 1 (BHV-1) glycoprotein D gene isolated in Indonesia Dewi Noor Hidayati; Eko Agus Srihanto; Tri Untari; Michael Haryadi Wibowo; Koichi Akiyama; Widya Asmara
Indonesian Journal of Biotechnology Vol 24, No 1 (2019)
Publisher : Universitas Gadjah Mada

Considering the increasing incidence of infectious bovine rhinotracheitis (IBR) in Indonesia, it was necessary to conduct a more in-depth study of bovine herpesvirus-1 (BHV-1) as the causative agent of IBR disease. Previous research reports indicate that the BHV-1 subtypes found in Indonesia are subtype 1.1. Currently, IBR field case detection in Indonesia still uses the serological method (ELISA), which has the potential to give false positive results and cannot explain the virus subtype. Other detection methods, such as viral isolation, take longer and require adequate resources. This study aimed to determine the BHV-1 subtypes of Indonesian isolates using molecular techniques. Nested PCR using two pairs of primers was successfully used to amplify the glycoprotein D (gD) gene. The gD gene fragment was cloned into the pGEM-T plasmid. Analysis of the gD gene sequence was subsequently carried out to determine the BHV-1 character of the Indonesian isolates. The results indicated that the isolates were different from the previous isolates, and had similarities (100%) with subtype 1.2 strain SP1777 and SM023.

PROFIL PLASMID Escherichia coli RESISTEN TERHADAP BEBERAPA ANTIBIOTIKA YANG DIISOLASI DARI PETERNAKAN AYAM KOMERSIAL Michael Haryadi Wibowo; Widagdo Sri Nugroho; Widya Asmara
Jurnal Sain Veteriner Vol 29, No 1 (2011): JUNI
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Penelitian ini bertujuan untuk mengetahui profil plasmid E.coli yang resisten terhadap ampisilin, streptomisin dan enrofloksasin. Delapan isolat E.coli yang telah diidentifikasi dan diuji sensitivitasnya terhadapketiga antibiotik tersebut, selanjutnya diisolasi plasmidnyas. Isolat E. coli dipupuk pada kaldu laktosa diinkubasi dalam shaker incubator pada suhu 370 C semalam. Sel dipanen dengan sentrifugasi 12.000 rpm, selama 5 menit sebanyak dua kali. Isolasi plasmid dilakukan dengan metode lisis alkali menggunakan larutan lisis I, II, dan III. Presipitasi plasmid dilakukan dengan 3 M Na asetat dan ethanol absolut. Profil plasmid dibaca pada agarosa 1%, setelah dilakukan elektroforesis menggunakan marker plasmid. Hasil penelitian menunjukkan profil plasmidDNA E. coli tersebut teramati sebagai pita-pita DNA yang berpendar oleh pendedahan sinar ultraviolet. Plasmid yang terisolasi mempunyai ukuran yang sangat besar atau mega plasmid yaitu terletak pada 4268 bp, 4873 bp, 5148 bp dan terletak diantara 5148 bp dan 21.226 bp. Masing-masing isolat E. coli memiliki jumlah plasmid yang bervariasi antar 1 sampai 3 plasmid DNA.Kata kunci: Escherichia coli, resistensi, profil plasmid

Isolasi dan Propagasi Agen Penyebab Penyakit dari Kasus Terdiagnosa Penyakit Infektious Laryngotracheitis (ILT) pada Telor Ayam Berembrio Michael Haryadi Wibowo; Widya Asmara
Jurnal Sain Veteriner Vol 20, No 2 (2002): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

EFEKTIVITAS PENGOBATAN PREPARAT KOMBINASI AMOKSISILIN DAN KOLISTIN SULFAT PADAKASUS INFEKSI BUATAN Escherichia coli PATOGEN PADA AYAM BROILER Michael Haryadi Wibowo; Surya Amanu
Jurnal Sain Veteriner Vol 27, No 1 (2009): JUNI
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Penelitian ini bertujuan untuk mengetahui efektifitas penggunaan preparat kombinasi amoksisilin dan kolistin suIfat pada kasus infeksi buatan Escherichia coli pada ayam broiler. Sebanyak 140 ekor ayam broiler dipelihara sejak umur satu hari menurut pemeliharaan standar yang lazim. Pada umur 21 hari, ayam tersebut dipisahkan menjadi duayaitu 118 ekor sebagai ayam perlakuan sedangkan sisanya 22 ekor sebagai kontrol. Ayam perlakuan diinfeksi E. coli patogenik isolat asal unggas (EC/Kls/4/02) secara intra peritoneal dosis 0,5 ml dari suspensi Mac Farland I. Segera setelah diinfeksi ayam tersebut diobati dengan preparat kombinasi amoksisilin dan kolistin dosis 1 gram per liter,diberikan selama 7 hari. Kelompok kontrol diinfeksi E. coli sebagaimana kelompok perlakuan tetapi tidak diobati. Respon pengobatan yang diamati adalah kesembuhan ayam yang teramati tanpa adanya gejala klinis, dan penampilan ayam paska pengobatan yang meliputi: rasio jumlah ayam terjual dari jumlah yang diinfeksi, berat badan dan tingkatkonversi pakan. Untuk membandingkan pengaruh pengobatan antara kelompok perlakuan dan kontrol diuji dengan analisis Chi- Square menggunakan program Student Edition of Statistic 4.0. Hasil penelitian ini menunjukkan bahwa prosentase ayam terjual sebanyak 86,4 %, dengan berat rerata pada umor 38 hari 1,48kg serta nilai efisiensipakan feedconvertion ratio 1,81. Secara statistik pengaruh pengobatan tersebut terdapat perbedaan yang bermakna pada tingkat signifikasi 0,5%, antara kelompok perlakuan dan kontrol. Berdasarkan data tersebut maka dapat disimpulkan bahwa respon pengobatan pada kasus infeksi buatan E. coli menggunakan preparat kombinasi amoksisilin dan kolistin sulfat,cukup baikuntuk mengatasi infeksi tersebut.Kata kunci: Escherichia coli, kombinasi antibiotik, potensi zooteknik ayam

PERBANDINGAN TITER ANTIBODI AYAM BROILER YANG DIVAKSIN PADA UMUR 7 DAN 14 HARI MENGGUNAKAN VAKSIN AVIAN INFLUENZA HETEROLOG SUBTIPE H5N2 Ukon Susetyo; Michael Haryadi Wibowo
Jurnal Sain Veteriner Vol 26, No 2 (2008): DESEMBER
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

The aim of this study was to compare the titer antibody respons induced by different vaccination programusing heterologous avian influenza H5N2 subtype at day seven and 14 in broiler chicken. A number of 120broiler chickens was divided into three groups, each consisted of 40 birds and kept since day old chicken instandard maintenance facilities. Group one as the treatment group was vaccinated at 7 days of age usingheterologous vaccine H5N2 subtype. The second group was vaccinated at 14 days of age using the samevaccine. The control group, received no vaccination. A number of24 birds in each group were randomly chosenand bled via brachialis vein at day 14, 21 and 28 post vaccination, respectively. The antibody titer in the serawas measured using hemaglutination inhibition (HI) test with homologous antigen H5N2 subtype. Data wereanalysed using a split-plot design of analysis of variance of 0,05, significance level. The titer induced by vaccinationat day-7 were 1,54; 15,92 and 6,58 HI unit, respectively. Meanwhile in the second group the titer obtained were1,29; 7,38 and 15 HI unit respectively. Antibody of the control group was negativ. Further analysis indicated thatthere was significance different antibody titer induced by vaccination between treatment groups, but whencompared between group of treatment, the result showed po significance different.Keys words: avian influenza. broiler chicken, heterologous vaccine, and antibody titer

Co-Authors Ade Erma Suryani Aditya Ahkami Pratomo AETH. Wahyuni Agnesia Endang Tri Hastuti Wahyuni Agnesia Endang Trihapsari Wahyuni Agus Eko Srihanto Agus Eko Srihanto, Agus Eko Agustina Dwi Wijayanti Ahmad Sofyan Akbar Agus Anshori Mussama Anastasia Endang Tri Hastuti Wahyuni Antasiswa Windraningtyas Rosetyadewi Arfian Rahma Shanti Arini Nurhandayani Aris Haryanto Bambang Sutrisno Bambang Sutrisno Bambang Sutrisno Charles Rangga Tabbu Charles Rangga Tabbu Claude Mona Airin Devi Yunita Sari Dewi Noor Hidayati Dini Wahyu Yudianingtyas Dito Anggoro Dito Anggoro Dito Anggoro Dyah Ayu Widiasih Eko Agus Srihanto Eko Agus Srihanto Eko Agus Srihanto Ferdinand Prayogo Cahyo Santoso Fidyah Fitrawati Fransiskus Trisakti Haryadi Hendra Herdian Heru Susetya I Nyoman Suarsana I Wayan Suardana I wayan Teguh Wibawan Ifah Khairunnizak Iwan Harjono Utama Khrisdiana Putri Khrisdiana Putri, Khrisdiana Koichi Akiyama Krisdiana Putri Lehgarubini Shanmuganathan Liza Angeliya Liza Angeliya Liza Angeliya Lusty Istiqomah Maria Anggita Marla Anggita Maxs Urias Ebenhaizar Sanam, Maxs Urias Medania Purwaningrum Mohammad Faiz Karimy Niken Respati Maharani Nur Khusni Hidayanto Nyoman Reishita Andriyani Okti Herawati Purnama Edy Santosa Radhian Fadiar Rahmat Setya Adji Ratna Ermawati Risang Aji Dewandaru Sarwo Edy Wibowo Sidna Artanto Sidna Artanto Sidna Artanto Sidna Artanto Sidna Artanto Sitarina Widyarini Siti Andarwarti Sugiyono Sugiyono Surya Amanu Surya Amanu Surya Amanu Surya Amanu Surya Amanu Surya Amanu Syarifah Alawiyah Tri Guntoro Tri Untari Tri Untari Tri Untari Tri Untari Ukon Susetyo Widagdo Sri Nugroho Widya Asmara Widya Asmara Widya Asmara Widya Asmara Widya Asmara Yuli Purwandari Kristiangingrum Yustina Yuni Suranindiyah

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