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Development of Recombinant Vaccine Using Herpesvirus of Turkey (Hvt) as Vector for Several Viral Diseases in Poultry Industry Hartawan, Risza
Indonesian Bulletin of Animal and Veterinary Sciences Vol 21, No 1 (2011)
Publisher : Indonesian Animal Sciences Society

Herpesvirus of turkey (HVT) has been utilised as live vaccine against Marekâs disease in poultry industry world-wide for many years. However, the potency of HVT is not limited on the Marekâs disease only. Along with rapid development of recombinant technique, the potency of HVT can be broaden for other diseases. As naturally apathogenic virus, HVT is a suitable candidate as vector vaccine to express important antigens of viral pathogens. Several researches have been dedicated to design HVT recombinant vaccine byÂ inserting gene of important virus, such as Marekâs disease virus (MDV), immuno bursal disease virus (IBDV), Newcastle disease virus (NDV) and Avian Influenza virus (AIV). Therefore, the future recombinant of HVT has been expected to be better in performance along with the improvement of recombinant technique. Key words: Herpesvirus of turkey, live vector vaccine, viral pathogens

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

Fenotipe Virus Avian Influenza (AI) Subtipe H5N1 Berbeda Karakter Genetik di Indonesia Dharmayanti, NLP Indi; Indriani, Risa; Hartawan, Risza; Ratnawati, Atik
JURNAL BIOLOGI INDONESIA Vol 10, No 2 (2014): Jurnal Biologi Indonesia
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v10i2.3031

In Indonesia, data on the phenotype of AI virus subtype H5N1 is very limited, due to the facilities for such testingshould be performed in a laboratory Biosafety level III. Additionally influenza virus has a high error rate duringtranscription of their genome has low RNA polymerase. High error rates generate quasispecies. In this study, wedid some H5N1 viruses infection with different genetic variations and conducted DNA sequencing in severalorgans. To determine the possible emergence detection of quasispecies in different organs and until the virus wasexcreted. We tested the hypothesis that the quasispecies might arise from virus that was infected to theexperimental animals. The results of this study showed that the AI virus subtype H5N1 had different phenotypes inanimal depending on their genetic character. AI virus subtype H5N1 killed chickens within 48-72 hoursdepending on the genetic character of the virus. New AI viruses of subtype H5N1 caused more severe organdamage severe than the character of the old AI viruses. Adaptation of virus in each organ also proved that the viruslikely create variations/quasispecies, which was different from the viral origin.Keywords: Phenotypes, avian influenza, H5N1, genetic character, quasispecies

SIRKULASI VIRUS AVIAN INFLUENZA SUBTIPE H5N1 DI PASAR TRADISIONAL DI JAWA TIMUR TAHUN 2012 [Circulation of the Avian Influenza Virus Subtype H5N1 at Traditional Markets of East Java in 2012] Hartawan, Risza; Dharmayanti, NLP Indi
BERITA BIOLOGI Vol 13, No 1 (2014)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Avian influenza virus subtype H5N1 outbreak has become endemic in Indonesia since 2003. The disease does not only cause immense economic losses but it also leads to significant fatality of human being. The existence of traditional markets including live bird trading is suspected to play important role in the spreading and evolution of the virus. The objective of this study was to identify the circulation of H5N1 virus at traditional markets of East Java in 2012 by using reverse transcriptase polymerase chain reaction (RT-PCR) test and virus isolation. As results, this study detected the presence of the H5N1 virus circulating in Gresik, Mojokerto, Lamongan and Surabaya in both of live birds and environmental samples. The successfulness of virus isolation indicated a potential transmission to other hosts, including to human. This study suggests that the improvement of the poultry trading system at traditional markets by implementing sanitation, hygiene and biosecurity is necessary to reduce the burden of virus contamination at the market environment.

FENOTIPE VIRUS AVIAN INFLUENZA (AI) SUBTIPE H5N1 BERBEDA KARAKTER GENETIK DI INDONESIA Dharmayanti, NLP Indi; Indriani, Risa; Hartawan, Risza; Ratnawati, Atik
JURNAL BIOLOGI INDONESIA Vol 10, No 2 (2014): Jurnal Biologi Indonesia
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v10i2.3031

In Indonesia, data on the phenotype of AI virus subtype H5N1 is very limited, due to the facilities for such testingshould be performed in a laboratory Biosafety level III. Additionally influenza virus has a high error rate duringtranscription of their genome has low RNA polymerase. High error rates generate quasispecies. In this study, wedid some H5N1 viruses infection with different genetic variations and conducted DNA sequencing in severalorgans. To determine the possible emergence detection of quasispecies in different organs and until the virus wasexcreted. We tested the hypothesis that the quasispecies might arise from virus that was infected to theexperimental animals. The results of this study showed that the AI virus subtype H5N1 had different phenotypes inanimal depending on their genetic character. AI virus subtype H5N1 killed chickens within 48-72 hoursdepending on the genetic character of the virus. New AI viruses of subtype H5N1 caused more severe organdamage severe than the character of the old AI viruses. Adaptation of virus in each organ also proved that the viruslikely create variations/quasispecies, which was different from the viral origin.Keywords: Phenotypes, avian influenza, H5N1, genetic character, quasispecies

IN SILICO ANALYSIS OF SMALL INTERFERING RNA TARGETING THE NUCLEOPROTEIN GENE OF INFLUENZA VIRUSES Hartawan, Risza; Ratnawati, Atik; Sendow, Indrawati; Dharmayanti, Ni Luh Putu Indi
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 11 No. 2 (2024)
Publisher : BRIN - Badan Riset dan Inovasi Nasional

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.55981/jbbi.2024.8521

Small interfering RNA (siRNA) is a promising therapeutic against viral infection, includ-ing Influenza viruses. However, the Influenza viruses have massive variants with high mutation rates. Therefore, the siRNAs could be futile against newly emerging viruses. Thus, this study aimed to analyze siRNAs targeting the nucleoprotein gene of Influen-za viruses. Using bioinformatic analyses, the siRNAs were simulated against 5 sub-types of Influenza viruses, including H1N1, H3N2, H5N1, H7N9, and H9N2. Bioinfor-matic tools for the folding structure of messenger RNA were utilized to select effective siRNA. As a result, 32 siRNA sequences targeting the nucleoprotein gene were identi-fied. The precision medicine concept seems applied to the siRNA treatment for the In-fluenza virus since each siRNA is effective in its respective virus target. Based on the nucleotide mismatch parameter, most siRNA does not have coverage for the multiple infections of all five subtypes of Influenza viruses, except for NP1089 and NP1496. Later, the secondary and tertiary structure analysis of messenger RNA demonstrated that siRNA has different circumstances in its RNA target position. Therefore, siRNA mapping based on the RNA folding structure approach provides a tool for selecting more effective sequences against Influenza virus infection. Both siRNA NP1089 and NP1496 were predicted to have similar effectivity in knocking down Influenza virus in-fection. Moreover, the cocktail application of siRNA treatment may be effective as an alternative strategy in matching co-infection of multiple Influenza virus subtypes.

DETECTION OF FIVE VIRUS INFECTIONS IN THE LAYER FARM WITH RUNTING-STUNTING SYNDROME IN SUKABUMI AND TANGERANG USING POLYMERASE CHAIN REACTION TECHNIQUE Hartawan, Risza; Indi Dharmayanti, Ni Luh Putu
Jurnal Kedokteran Hewan Vol 11, No 2 (2017): June
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v11i2.5400

The objective of this study was to identify the five different viral infections including avian influenza, Newcastle disease, avian reovirus, avian encephalomyelitis, and Mareks disease in the runting-stunting syndrome outbreak in several commercial layer farms in Sukabumi and Tangerang in November 2014 using polymerase chain reaction technique. As results, this study identified mix infection of three viruses in the field samples, including Newcastle disease, reovirus, and avian encephalomyelitis; however, it was negative for avian influenza and Mareks disease viruses. Subsequently, the inoculation of several samples into embryonated chicken eggs confirmed the growth of these three viruses. As a consequence, disease control management should be conducted in the affected farms by implementing effective biosecurity and vaccination program.

APOPTOSIS STUDY OF INDONESIAN AVIAN INFLUENZA VIRUS SUBTYPE H5N1 IN MADIN-DARBY CANINE KIDNEY CELLS Dharmayanti, NLP Indi; Ukhti, Dwi Rillah; Syamsiah, Farida; Hartawan, Risza
Jurnal Kedokteran Hewan Vol 11, No 1 (2017): March
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v11i1.5378

This study aimed to determine the ability of highly pathogenic avian influenza virus (HPAI) virus subtype H5N1 originated from Indonesia to induce apoptosis in Madin-Darby Canine Kidney (MDCK) cells. Three HPAI virus subtype H5N1 isolates with different genetic characteristic namely A/Bird/Bali1/2011, A/Chicken/East Java/BwiI2/2010 and A/Chicken/West Java/1074/2003, were cultured in MDCK cells. Apoptosis was identified by deoxyribonucleic acid (DNA) fragmentation of infected MDCK cells using Apoptotic DNA Ladder Kit. The results showed that all three HPAI virus isolates used in this study did not able to induce apoptosis in the MDCK cells within 5 to 72 hours post infection.

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