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All Journal Jurnal Ilmu Ternak dan Veteriner WARTAZOA Indonesian Bulletin of Animal and Veterinary Sciences BERITA BIOLOGI JURNAL BIOLOGI INDONESIA

RISZA HARTAWAN

Indonesia Research Center for Veterinary Science Indonesian Agency for Agricultural Research and Development Jalan R.E. Martadinata No. 30, PO Bx 52 Bogor 16114, West Java, Indonesia

Author-ID : 276889

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Veterinary

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Articles

Title

Identification of Mardivirus Serotypes Circulating in Poultry Farms in Sukabumi and Cianjur District, West Java, 2011 using Multiplex Polymerase Chain Reaction (mPCR) Approach Hartawan, Risza; NLPI, DharmayantI
Indonesian Journal of Animal and Veterinary Sciences Vol 18, No 4 (2013)
Publisher : Indonesian Animal Sciences Society

Three serotypes of Mardivirus had been circulating in the farm environments, these being Marekâs disease virus serotype 1 (MDV-1), Gallid hepesvirus 3 (GaHV3) and herpesvirus of turkey (HVT). However, only MDV-1 poses a significant hazard to the poultry farm. The virus causes a neoplastic syndrome that inflicting severe economic loss to the affected farms. Although vaccination has successfully reduced the frequency and severity of outbreaks, the threat does not disappear since several more pathogenic strains have evolved, and these can overcome protection by vaccination. The aim of this study was to investigate the circulation of three Mardivirus serotypes in commercial poultry farms in Sukabumi and Cianjur district using mPCR approach for the feather samples. A low prevalence of these three serotypes was detected. However, the practice of vaccinating using live attenuated MDV-1 caused difficulty in the investigation. Differentation between virulent field strains and CVI988 vaccine strain using the 132 bp repeat motif attenuation marker within the terminal and inverted repeats flanking the unique long region generated an ambiguous result. Thus, other approaches are required to address this issue, such as selection of other markers, restriction fragment length polymorphism (RFLP), high-resolution melt curve analysis (HRM) and gene sequencing. Key Words: Mardivirus serotype, MDV-1, GaHV3, HVT, multiplex PCR

In vitro expression of native H5 and N1 genes of avian influenza virus by using Green Fluorescent Protein as reporter Hartawan, Risza; Robinson, K.; Mahony, T.; Meer, J.
Indonesian Journal of Animal and Veterinary Sciences Vol 16, No 3 (2011)
Publisher : Indonesian Animal Sciences Society

The hemagglutinin and neuraminidase are important immunogen of avian influenza virus that are suitable for recombinant experimentation. However, both genes have been experienced rapid mutation resulting in diverse variety of genotypes. Hence, gene expression in recombinant systems will be difficult to predict. The objective of the study was to examine expression level of H5 and N1 genes from a field isolate by cloning the genes into expression vector pEGFP-C1. Two clones respresenting fulllength of H5 and N1 gene in plasmid pEGFP-C1 were transfected into chicken embryo fibroblasts (CEF), rabbit kidney (RK13) and African green monkey kidney (VERO) cells using Lipofectamine âPlusâ reagent. The experiment showed level of gene expression in the VERO cell was higher than in the RK13 and CEF cells. Observations using fluorescent microscopy and Western blotting revealed that the N1 gene was expressed better in all cells compared to the H5 gene. Key Words: H5N1 Virus, Hemagglutinin, Neuraminidase, Gene Expression, Green Fluorescent Protein

Characterisation of the H5 and N1 genes of an Indonesian highly pathogenic Avian Influenza virus isolate by sequencing of multiple clone approach Hartawan, Risza; Robinson, Karl; Mahony, Timothy; Meers, Joanne
Indonesian Journal of Animal and Veterinary Sciences Vol 15, No 3 (2010)
Publisher : Indonesian Animal Sciences Society

Hemagglutinin and neuraminidase are the main antigenic determinants of highly pathogenic avian influenza (HPAI) virus. The features of these surface glycoproteins have been intensively studied at the molecular level. The objective of this research was to characterise the genes encoding these glycoproteins by sequencing of multiple clones. The H5 and N1 genes of isolate A/duck/Tangerang/Bbalitvet-ACIAR-TE11/2007 were each amplified in one or two fragments using reverse transcriptase-PCR (RT-PCR), and subsequently cloned into pGEM-T Easy TA cloning system. The sequencing result demonstrated high homology between respective clones but with several variations that were identified as single nucleotide polymorphisms (SNPs). A total of 1,707 base pair and 1,350 base pair of H5 and N1 genes respectively were successfully assembled from multiple clones containing the genes of interest. The features of both H5 and N1 genes from this isolate resemble the typical characteristics of Indonesian strains of H5N1 virus from sub-clade 2.1.3. Key Words: Avian Influenza, Characterization, Gene Cloning, Hemagglutinin, Neuraminidase

Co-Authors Atik Ratnawati DharmayantI NLPI J. Meer JOANNE MEERS K. Robinson KARL ROBINSON NLP Indi Dharmayanti NLPI Dharmayanti RISA INDRIANI T. Mahony TIMOTHY MAHONY

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