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All Journal Jurnal Sain Veteriner Indonesian Journal of Biotechnology Journal of Tropical Biodiversity and Biotechnology JURNAL KAJIAN VETERINER Jurnal Nasional Teknologi Terapan Jurnal Kedokteran Hewan

Aris Haryanto

Fakultas Kedokteran Hewan Universitas Gadjah Mada

Author-ID : 2760530

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Civil Engineering, Building, Construction & Architecture Computer Science & IT Control & Systems Engineering Electrical & Electronics Engineering Environmental Science Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Public Health Veterinary

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Diagnosis and molecular characterization of Anaplasma platys in dog patients in Yogyakarta area, Indonesia Muh. Disna Faizal; Aris Haryanto; Ida Tjahajati
Indonesian Journal of Biotechnology Vol 24, No 1 (2019)
Publisher : Universitas Gadjah Mada

Anaplasma platys is a tick-borne, Gram-negative bacterium that causes anaplasmosis, a companion vector-borne disease impacting dogs. Information on this disease remains limited in Indonesia. Its symptoms are not specific, so molecular analysis is required for a rapid and accurate diagnosis. GroEL is an essential gene commonly used for classification and species identification of many groups of bacteria, including Anaplasma spp. In this study, a molecular diagnosis of anaplasmosis based on the groEL gene sequence was conducted using PCR. In addition, the genetic diversity of Anaplasma platys in infected dogs was determined. Blood samples were collected from 51 dogs suspected of anaplasmosis from Prof. Dr. Soeparwi Animal Hospital, animal clinics, and pet shops in the Yogyakarta area, Indonesia, based on anamnesis, histories of tick infestations, and clinical symptom examinations. DNA extraction and PCR targeting the groEL gene were performed, followed by sequencing. Phylogenetic tree analysis and construction were carried out using the BLAST and MEGA programs. Positive PCR sample results (amplicon length of 624 bp) were found in 6 of 51 dogs. Samples A1 (KHJ/C2), A2 (KHJ/A2), A3 (KSK/L), A4 (KHJ/L), and A5 (KNP/M2) had close ties to Anaplasma platys (AF478129.1) from GenBank. Phylogenetic analysis showed a very high homology value (100%) and bootstrap value of 100%. It can be concluded that there was no genetic diversity in the Anaplasma platys found in infected dogs in the Yogyakarta area.

Molecular bird sexing of sulphur‐crested cockatoo (Cacatua galerita) by polymerase chain reaction method Diana Savitri; Irhamna Putri; Warih Pulung Nugrahani; Medania Purwaningrum; Aris Haryanto
Indonesian Journal of Biotechnology Vol 26, No 1 (2021)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.54611

Sex identification of endangered and protected birds in captivity is very important for conservation programs. Half of the world’s bird species are monomorphic, where male and female are difficult to distinguished morphologically, including cockatoos. Sex identification using molecular bird sexing is more accurate and applicable because it directly targets the sex chromosomes. The purpose of this study was to determine the sex of Sulphur‐crested cockatoo (Cacatua galerita) by detecting differences in the intron size of the chromodomain helicase DNA‐binding 1 (CHD1) gene on the Z and W chromosomes by polymerase chain reaction (PCR) method and to compare of plucked feathers and blood samples as DNA sources for molecular bird sexing. DNA was extracted from feather and blood samples from four C. galerita. Extracted DNA was amplified on the CHD1 gene by PCR method with P2, MP, and NP primers, which were visualized using agarose gel 1.5% under UV transilluminator with a wavelength of 280 nm. The resulting PCR product was detected at 392 bp for the CHD1 Z gene segment and 297 bp for CHD1 W gene segments, where males showed a single DNA band (ZZ) and females showed a double DNA band (ZW). Four C. galerita were 100% successfully determined, consisting of one female and three males. Electrophoresis results showed DNA bands from blood samples were thicker and brighter than DNA bands from feather samples.

In vitro expression of the recombinant fusion protein of Newcastle disease virus from local Indonesian isolates by using a cell-free protein expression system Aris Haryanto; Hevi Wihadmadyatami; Nastiti Wijayanti
Indonesian Journal of Biotechnology Vol 25, No 2 (2020)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.54703

The aim of this work was the in vitro expression of the recombinant fusion (F) protein of Newcastle disease virus (NDV). The pBT7-N-His-Fusion-NDV expression plasmid which carries the recombinant F protein encoding gene from local Indonesian isolates, was prepared and transformed into E. coli BL21 (DE3). To detect bacterial colonies carrying the recombinant plasmid, a restriction endonuclease analysis was performed using the EcoRI restriction endonuclease. These results showed that the pBT-N-His-Fusion-NDV plasmid was successfully isolated with a size of 4.601 bp, and three recombinant plasmids carrying the gene coding for the recombinant F protein of NDV were obtained. Selected recombinant plasmids were then in vitro by using a cell-free protein expression system followed by visualization of the recombinant F protein on a 12% SDS-PAGE gel both by Coomassie Brilliant Blue staining and Western blotting. Recombinant F protein was successfully in vitro expressed by using a cell-free protein expression system as indicated by a specific single protein band with a molecular mass of 25.6 kDa.

PREPARATION OF LYMPHOCYTE CULTURE CELL FROM PERIPHERIAL BLOOD OF NASOPHARYNGEAL CARCINOMA PATIENTS Aris Haryanto; Sofia Mubarika; Nastiti Wijayanti
Jurnal Sain Veteriner Vol 18, No 1&2 (2000)
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.8239

Lymphocyte is leukocyte component that difficult to culture in vitro. Several viruses need lymphocytes as host cell in order to proliferate and growth in this media such as Epstein-Barr virus (EBV). This virus was associated with malignant disease like nasopharyngeal carcinoma (NPC). The objectives of this research are to develop and to prepare lymphocyte cell culture as material source of DNA for molecular analysis of virus. Peripheral blood was collected from NPC patients which is histopatologically and serologically positive of EBV. Lymphocytes were separated from the other blood components using ficcol-histopaque. Lymphocytes were diluted using RPMt medium then they were cultivated into 96 microwell plate with concentration of 106 cell/ml. The medium consist of 10% FBS, RPMI, Penstrep and FK 506. Culture of lymphocytes were incubated in 5% CO2 at 37°C. The lymphocyte cultures developed and grew confluendy during the first week. Only B cells whith EBV would be well establish. After 50 cell generations, lymphocytes were transformed and immortalized to be lymphoblastoid cell fine (LCL).

Molecular Bird Sexing of Tanimbar Cockatoos (Cacatua goffiniana) by Using Polymerase Chain Reaction Method Ratu Fresa Khoirotunnisa Hidayat; Diana Savitri; Irhamna Putri; Warih Pulung Nugrahani; Aris Haryanto
Journal of Tropical Biodiversity and Biotechnology Vol 6, No 2 (2021): August
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jtbb.59997

This study aimed to determine the sex of Tanimbar Cockatoo (Cacatua goffiniana) birds by amplifying Chromodomain Helicase DNA-binding-1 (CHD-1) gene on Z and W sex chromosomes as well as to compare the quality of DNA extraction and PCR amplification products from samples derived from peripheral blood and plucked feathers. This work used five C. goffiniana birds which were collected from the Wildlife Rescue Center (WRC) in Pengasih, Kulon Progo, Yogyakarta. From each C. goffiniana, feather samples were collected by plucking feathers on the ventral wing and peripheral blood samples were taken by cutting their nails and collecting the blood into microhematocrit tubes containing heparin. The next stage was DNA extraction and DNA amplification on the CHD-1 gene using the PCR method by NP, P2, and MP primer pairs. Then, products of DNA extraction and PCR amplification were electrophoresed on 1.5% agarose gel and visualized under a UV light transilluminator with a wavelength of 260 nm. The visualization showed that samples from peripheral blood generated clearer DNA fragments compared to plucked feathers. Two of the five samples were male C. goffiniana and the other three samples were females. In the male Tanimbar Cockatoo was amplified a single DNA fragment of the Z sex chromosome in size of 300 bp, whereas in the female C. goffiniana was amplified double DNA fragments of Z and W sex chromosomes in size of 300 bp and 400 bp respectively. The DNA quality showed that the DNA quality from peripheral blood samples were better in quality than the DNA collected from plucked feather samples.

Deteksi Molekuler Gen Fusion (F) dan Analisis Perbandingan Beberapa Enzim Restriksi sebagai Penentu Patotipe Virus Newcastle Disease Medania Purwaningrum; Verawati Verawati; Aris Haryanto
Jurnal Nasional Teknologi Terapan (JNTT) Vol 2, No 3 (2018): NOVEMBER
Publisher : Penelitian dan Pengabdian Kepada Masyarakat Sekolah Vokasi Universitas Gadjah Mada

Newcastle disease (ND) is a contagious viral disease caused by Avian Paramyxovirus Serotype-1 (APMV-1). This viral infection is responsible for devastating outbreak by attacking nerve, respiration, and also digestive system. This disease often followed with decreasing of eggs production and also responsible for economic losses in the poultry industries around the globe. The main goal was to differentiate virulent or avirulent strain of ND virus from F gene, which is the virulent marker of ND virus, by Reverse Transcription Polymerase Chain Reaction (RT-PCR) and Restrion Enzyme Analysis using BamH1, Hin 1l, and Apa 1. Ten ND virus samples came from Animal Disease Investigation Center (ADIC) Wates virus collection, collected from field case in 2012-2013. Newcastle disease virus was collected by extraction from the samples. The RNA product of extraction were used as a template for amplification in RT-PCR. The target of RT-PCR amplification was F gene. The results indicated positive reaction due to existing of DNA fragment band in size of 767 bp. RT-PCR and Restrion Enzyme Analysis can be used as tool to determine the pathotype of ND virus showed different restriction visualized by gel agarose electrophoresis.

Perubahan Patologis Hepar Akibat Cemaran Aflatoksin B1 Pada Pakan Ayam Pedaging Komersial Di Kota Kupang Devi Y. J. A. Moenek; Aris Haryanto; Charles Rangga Tabu
JURNAL KAJIAN VETERINER Vol 4 No 1 (2016): Jurnal Kajian Veteriner
Publisher : FAKULTAS KEDOKTERAN HEWAN UNIVERSITAS NUSA CENDANA

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35508/jkv.v4i1.1010

Aflatoxin B1 is a secondary metabolite of Aspergillus flavus, Aspergillus parasiticus, and Penicillium puberulum fungus, which is frequently found as contaminants of feed/raw materials of poultry feed. Such compounds have toxic and carcinogenic effects that can cause damage to various tissues/organs, which can further decrease the performance of broiler and cause various degrees of immunosuppressive effects. This study was designed to evaluate the most common pathological lesion in the liver of commercial broiler after consumption of aflatoxin contamination feed in Kupang City of East Nusa Tenggara Province. The research was conducted on 10 broiler farms in Kupang City. Samples of liver were taken for further processing according to the staining method of haematoxylin and eosin (H&E). The results of pathological examination of tissues were analyzed descriptively. Based on the results of this experiment, it can be concluded that pathological examination on the liver showed an early-stage of liver cirrhosis, atrophy of bursa fabricius, thymus and lien due to the necrosis and depletion of lymphocytes.

PENENTUAN SUBTIPE VIRUS AVIAN INFLUENZA DENGAN METODE SINGLE STEP MULTIPLEX REVERSE TRANSCRIPTASE- POLYMERASE CHAIN REACTION (RT-PCR) ISOLAT ASAL PROVINSI ACEH Teuku Zahrial Helmi; Rini Widayanti; Aris Haryanto
Jurnal Kedokteran Hewan Vol 8, No 1 (2014): March
Publisher : Universitas Syiah Kuala

Tujuan dari penelitian ini adalah mengidentifikasi keberadaan gen M, H5, dan N1 virus avian influenza (AI) melalui metode single step multiplex reverse transcriptase-polymerase chain reaction (RT-PCR), sebagai acuan untuk peneguhan diagnosis secara molekuler virus AI di Provinsi Aceh. Penelitian ini menggunakan 11 isolat virus AI asal Provinsi Aceh yang diperoleh dari Laboratorium Balai Penyidikan dan Pengujian Veteriner (BPPV) Regional I Medan di Sumatera Utara dari tahun 2006-2008. Penelitian dilakukan di Laboratorium Virologi BPPV Regional I di Medan, Sumatera Utara. Amplifikasi terhadap gen matriks (M) virus AI menggunakan metode simplex RT-PCR. Hasil simplex RT-PCR terhadap gen M diperoleh 10 isolat yang menunjukkan pita deoxyribonucleic acid (DNA) pada 276 bp dan satu isolat yang tidak muncul, kemudian dilanjutkan dengan metode single step multiplex RT-PCR menggunakan pasangan primer gen penyandi protein N1, H5, dan M. Produk PCR 131 bp (N1), 189 bp (H5), dan 276 bp (M) muncul sebagai hasil elektroforesis dari semua isolat virus AI. Semua virus AI yang mewabah dari tahun 2006-2008 di Provinsi Aceh termasuk ke dalam virus influenza A subtipe H5N1.

ANALISIS FILOGENETIK ISOLAT VIRUS AVIAN INFLUENZA SUBTIPE H5N1 ASAL PROVINSI ACEH Teuku Zahrial Helmy; Rini Widayanti; Aris Haryanto
Jurnal Kedokteran Hewan Vol 6, No 1 (2012): March
Publisher : Universitas Syiah Kuala

Penelitian ini bertujuan melakukan studi filogenetik virus AI tipe A subtipe H5N1 isolat asal Provinsi Aceh yang dapat memberikan informasi secara molekuler tentang kekerabatan antar isolat virus AI tipe A subtipe H5N1. Sebanyak 11 isolat virus AI asal Provinsi Aceh yang dikoleksi oleh Laboratorium Virologi Balai Penyidikan dan Pengujian Veteriner (BPPV) Regional I di Medan, Sumatera Utara selama kurun waktu 2007-2008 digunakan dalam penelitian ini. Metode penelitian meliputi preparasi sampel isolat virus, ekstraksi RNA, amplifikasi gen H, elektroforesis DNA hasil amplifikasi, sekuensing, dan analisis data hasil sekuensing dengan software MEGA 4.0. Hasil sekuensing diketahui bahwa daerah yang teramplifikasi sebesar 191 bp. Hasil allignment dari nukleotida 191 bp ditemukan urutan nukleotida yang menyandi 5 asam amino yang berbeda pada posisi asam amino ke-493, 498, 499, 504, dan 515 antara isolat virus AI subtipe H5N1 asal Provinsi Aceh selama tahun 2007-2008 dengan isolat dari pembanding dari gene bank. Kontruksi pohon filogenetik menunjukkan bahwa virus AI subtipe H5N1 asal Provinsi Aceh tahun 2007-2008 mengelompok secara terpisah dari isolat Indonesia yang lain, tetapi masih dalam klaster virus AI subtipe H5N1 Indonesia.

DIAGNOSIS CEPAT VIRUS AVIAN INFLUENZA TIPE A SUBTIPE H5 DARI SPESIMEN LAPANGAN DENGAN METODE ONESTEP SIMPLEX RT-PCR Aris Haryanto; Duhita Andinita; Sri Handayani Irianingsih; Dini Wahyu Yudianingtyas
Jurnal Kedokteran Hewan Vol 6, No 1 (2012): March
Publisher : Universitas Syiah Kuala

Virus avian influenza (AI) merupakan virus dengan materi genetik RNA single-stranded sense negatif, beramplop yang termasuk dalam famili Orthomyxoviridae. Onestep simplex reverse transcriptase-polymerase chain reaction (RT-PCR) merupakan salah satu metode diagnosis yang dapat diandalkan untuk mendeteksi virus AI. Tujuan dari penelitian ini adalah untuk menerapkan metode diagnosis cepat virus AI pada spesimen lapangan secara langsung dari pasar unggas berdasarkan amplifikasi RT-PCR gen M dan H5 dengan metode yang berbasis onestep simplex RT-PCR tanpa melalui proses inokulasi dan propagasi virus AI dalam telur ayam berembrio (TAB). Sebanyak 35 sampel spesimen lapangan dari swab trakea unggas yang berasal dari pasar unggas di Terban, Kotamadya Yogyakarta digunakan dalam penelitian ini. Amplifikasi DNA secara onestep simplex RT-PCR pada gen matriks (M) dilakukan terhadap seluruh sampel. Pada sampel-sampel yang menunjukkan hasil positif pada amplifikasi gen M kemudian dilakukan amplifikasi RT-PCR secara lebih lanjut untuk gen H5 virus AI. Produk hasil amplifikasi RT-PCR gen M dan H5 divisualisasikan menggunakan elektroforesis gel agarose konsentrasi 1% dengan pewarnaan SYBR® Safe DNA Gel Staining. Hasil amplifikasi RT-PCR gen M menunjukkan bahwa dari 35 sampel diperoleh 8 sampel positif terinfeksi virus AI tipe A yang ditunjukkan dengan adanya fragmen DNA sebesar 200 bp, sedangkan hasil amplifikasi gen H5 sebanyak 5 dari 8 sampel-sampel tersebut merupakan virus AI tipe A subtipe H5 yang ditunjukkan dengan adanya fragmen DNA sebesar 545 bp. Diagnosis cepat virus AI tipe A subtipe H5 secara langsung dari spesimen lapangan di pasar unggas dapat dilakukan dengan metode onestep simplex RT-PCR, namun metode diagnosis tersebut tidak dapat mendeteksi keberadaan virus AI dalam sampel yang virusnya terlalu sedikit.

Co-Authors Charles Rangga Tabu Diana Savitri Diana Savitri Dini Wahyu Yudianingtyas Duhita Andinita Hevi Wihadmadyatami Ida Tjahajati Irhamna Putri Irhamna Putri Medania Purwaningrum Moenek, Devi Y.J.A. Muh. Disna Faizal Nastiti Wijayanti Nastiti Wijayanti Ratu Fresa Khoirotunnisa Hidayat Rini Widayanti Sofia Mubarika Sri Handayani Irianingsih Teuku Zahrial Helmi Teuku Zahrial Helmy Verawati Verawati Warih Pulung Nugrahani Warih Pulung Nugrahani

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