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Detection of antibody responses by using haemagglutination inhibiton test and the protection titer of avian influenza virus H5N1 subtype Indriani, Risa; Dharmayanti, N.L.P.I; Wiyono, A; ., Darminto; Parede, L
Indonesian Journal of Animal and Veterinary Sciences Vol 9, No 3 (2004)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (166.554 KB) | DOI: 10.14334/jitv.v9i3.410

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Study on the detection of antibody responses using haemagglutination inhibition (HI) test and the protection titer to Avian influenza (AI) virus H5N1 subtype local isolate has been conducted at the Research Institute for Veterinary Science (RIVS). A total number of 50 village chicken (10 chicken served as un-injected controls) and 30 quail were injected intramuscularly with inactivated virus of AI H5N1 subtype local isolate. Serum samples were collected 3 weeks after injection and were tested using haemagglutination inhibition tests. The correlation between antibody titer and its protection to AI virus H5N1 local isolate were measured by challenging the birds with AI virus H5N1 local isolate The HI test was then used to determine field serum samples. A total number of 48 village chicken from three (3) Districts (Bekasi, Tangerang and Bogor) and 96 quails from two (2) farms in District of Sukabumi which were all vaccinated with commercial AI adjuvant vaccine were sampled. The study revealed that village chicken and quails showed antibody responses after 3 weeks vaccination and that titer of ≥ 3 log 2 was able to protect chicken and quails when they were challenged with local isolate virus. Based on this result, village chicken field samples from Districts of Tangerang, Bekasi and Bogor showed antibody titer which will protect 50, 100 and 85% of the flocks respectively. While quail field samples from Farm I and Farm II in District of Sukabumi showed antibody titer which will protect 60-100% and 0-80% of the flocks respectively. It is concluded that the study has successfully measured antibody titer to AI virus H5N1 subtype which protect village chicken and quails from local isolate virus challenge so that the results will be used to analyze field serum samples after vaccination program to eradicate AI from Indonesia.   Key words: Antibody responses, haemagglutination inhibition test, protection titer, AI virus H5N1subtype
Identification of avian influenza virus of Indonesian isolates by reverse transcriptase polymerase chain reaction (RT-PCR) method Dharmayanti, N.L.P.I.; Damayanti, R; Wiyono, A; Indriani, R; ., Darminto
Indonesian Journal of Animal and Veterinary Sciences Vol 9, No 2 (2004)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (233.427 KB) | DOI: 10.14334/jitv.v9i2.420

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An outbreak of avian influenza in Indonesia was reported at the first time at the beginning of September 2003 causing high mortality among poultry population especially commercial layer chicken farms in Java, Sumatra and Bali islands. From the outbreaks highly pathogenic avian infuenza viruses have been isolated and characterized by rapid, HA, HI and AGP tests. However, these isolates are still needed to be further molecularly characterized. The aim of this study is to identify by further subtyping the avian viruses by means of RT-PCR using Matrix, H7 and H5 primers. The study reveals that the RT-PCR using Matrix primer amplified a 200-300 basepairs (bp) Jawa Timur isolates were collected from East Java, while Jawa Barat isolates were from West Java. The RT-PCR using H7 primers did not amplify any product, while H5 primer amplified a 500-600 bp product from the isolates. It is concluded that the outbreak of poultry disease in East and West Java was caused by an avian influenza H5 subtype.   Key words: Identification, avian influenza virus, RT-PCR, H5 subtype
Characterisation of enzymatic activities of H5N1 influenza virus Tarigan, Simson; Indriani, Risa; ., Darminto
Indonesian Journal of Animal and Veterinary Sciences Vol 12, No 2 (2007)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (195.159 KB) | DOI: 10.14334/jitv.v12i2.554

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One of the two glycoproteins projected from the surface of the influenza virus is identified as neuraminidase. This enzyme enables the virus to spread in the host, and therefore it plays vital roles in the viral pathogenicity. From the viewpoint of disease control, neuraminidase is used as the target for the development of anti-flu drugs, and for the development of diagnostic test to differentiate infected from vaccinated animals (DIVA). Since the roles of the enzyme are very important, information regarding the characteristics and the procedure to measure its activity, which is the purpose of this study, is essential. The optimum incubation time of the neuraminidase-substrate (fetuin) reaction and the optimum pH of the buffer were determined. The stability of the enzyme against heating, supplementation or chelating of calcium ion, and b-propiolactone treatment were analysed. This study showed that neuraminidase from H5N1-influenza virus was, in regards to the characteristics investigated in this study, was comparable to that from Clostridium perfringens. The optimum incubation time for the viral and Clostridial neuraminidases were 60 and 30 minutes, respectively; whereas, the optimum pH for both neuraminidase was 6-7. At pH 8, both neuraminidase were inactive. Supplementation of calcium ion tended to increase activity but chelating of the cation did not have any observable effects. Treatment with 0.2% b-propiolactone for 6 hours reduced the activity, whereas heating at 60°C for 60 minutes abolished all activity. Since inactivation by b-propiolactone is partially only, neuraminidase assay could be performed safely in ordinary laboratories using b-propiolactone-treated-influenza virus, rather than the life virus. The thermolabile nature of the enzyme will complicate any attempt to purify the enzyme. Key Words: H5N1, Neuraminidase, Stability, Thiobarbituric Assay
Genetic characteristic of protein membran of avian influenza viruses H5N1 subtype Dharmayanti, N.L.P Indi; Hewajuli, D.A; Ratnawati, A; Indriani, R; ., Darminto
Indonesian Journal of Animal and Veterinary Sciences Vol 15, No 3 (2010)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (195.271 KB) | DOI: 10.14334/jitv.v15i3.662

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In 2006-2008 there were findings about the antigenic drift on AI virus due to vaccination and the AI H5N1 subtype viruses which was similar to H5N1 viruses in human. The findings indicated that the AI viruses continue and undergoing to mutate and try to adapt with their environment.  The objective of this study was to characterize the mutation of recent AI viruses (2009) on the membran protein namely Hemagglutinin (HA), Neuraminidase (NA) and Matrix 2 (M2). In this study RT-PCR – sequencing methods and genetic analysis for the protein membran of AI viruses were used. Result revealed that there were specific mutation belong to AI 2009 viruses on HA and NA protein such as AI virus mutation in 2008 which was isolated from backyard chicken. The mutations were non synonimous and not caused by immunological pressure. Furthermore, M2 analysis indicated that the viruses were resistant to amantadine. Key Words: Mutation, AI Subtype H5N1 Viruses, Membran Protein
Isolation and characterization of virus of highly pathogenic avian influenza H5 subtype of chicken from outbreaks in Indonesia Wiyono, Agus; Indriani, R; Dhamaryanti, N.L.P.I; Damayanti, R; Parede, L; Syafriati, T; ., Darminto
Indonesian Journal of Animal and Veterinary Sciences Vol 9, No 1 (2004)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (515.934 KB) | DOI: 10.14334/jitv.v9i1.429

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A study on the isolation and characterization of Highly Pathogenic Avian Influenza of chicken from outbreaks in Indonesia was conducted at Indonesian Research Institute for Veterinary Science. Outbreaks of avian disease had been reported in Indonesia since August 2003 affecting commercial layer, broiler, quail, and ostrich and also native chicken with showing clinical signs such as cyanosis of wattle and comb, nasal discharges and hypersalivation, subcutaneous ptechiae on foot and leg, diarre and sudden high mortality. The aim of this study is to isolate and characterize the causal agent of the disease. Samples of serum, feather follicle, tracheal swab, as well as organs of proventriculus, intestine, caecal tonsil, trachea and lungs were collected from infected animals. Serum samples were tested haemaglutination/haemaglutination inhibition to Newcastle Disease and Egg Drop Syndrome viruses. Isolation of virus of the causal agent of the outbreak was conducted from samples of feather follicle, tracheal swab, and organs using 11 days old specific pathogen free (SPF) embryonated eggs. The isolated viruses were then characterised by agar gel precipitation test using swine influenza reference antisera, by haemaglutination inhibition using H1 to H15 reference antisera, and by electron microscope examination. The pathogenicity of the viruses was confirmed by intravenous pathogenicity index test and its culture in Chicken Embryo Fibroblast primary cell culture without addition of trypsin. The study revealed that the causative agent of the outbreaks of avian disease in Indonesia was avian influenza H5 subtype virus based upon serological tests, virus isolation and characterization using swine influenza reference antisera, and electron microscope examination. While subtyping of the viruses using H1 to H15 reference antisera suggested that the virus is very likely to be an avian influenza H5N1 subtype virus. The pathogenicity test confirmed that the viruses are highly pathogenic to experimental animals. It is concluded that the causative agent of the outbreaks of avian disease in Indonesia was avian influenza H5 subtype virus. The result has been the basis of further study such as development serological tests and vaccine production. The decission of Indonesian Government to conduct vaccination program using homolog vaccine in order to control the disease is regarded as the correct choice. However, it should be accompanied by conducting surveillance and monitoring of the disease as well as the possibility of mutation of virus. The program should be coordinated nationally.   Key words: Virus isolation, characterization, chicken, outbreak, highly pathogenic avian influenza (HPAI), H5 subtype, Indonesia
Development of a nested PCR for detection of Bovine herpesvirus-1 (BHV-1) in bovine nasal secretion and semen Saepulloh, Muharam; Adjid, R.M Abdul; T. Wibawan, I Wayan; ., Darminto
Indonesian Journal of Animal and Veterinary Sciences Vol 13, No 2 (2008)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (283.13 KB) | DOI: 10.14334/jitv.v13i2.609

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A nested polymerase chain reaction (nPCR) assay for detection of Bovine herpesvirus-1 (BHV-1) in bovine semen and nasal secretions was successfully developed. The nested Polymerase Chain Reaction was based on external and internal primers from the viral gD glycoprotein gene. This nPCR assay was 1000-fold more sensitive than using PCR external primer. The nested PCR has a detection limit as low as 5 ag/ml pure BHV-1 DNA and 100,75 TCID50/500 mL BHV-1 infected cells. On the other hand,  PCR using external primer had detection limit of about 5 fg/ml pure BHV-1 DNA. Specificity studies showed that nPCR could only detect BHV-1, whereas BHV-4, PRV, PI-3 and BRSV can not be detected. In addition, nPCR was also capable in detecting BHV-1 in nasal secretion samples from animal without clinical signs. A total of 405 samples consisted of 381 nasal secretion and 24 fresh semen samples have been tested with the nPCR. The result revealed that from 381 nasal secretion samples tested, 14 samples showed to be positive (3.68%), consisting of 13 out of 294 (4.42%) nasal secretion samples collected from Pangalengan West Java, and 1 out of 87 (1.54%) samples collected from Bogor. Furthermore, 2 out of 11 (18.18%) extended semen samples collected from Bogor and 2 out of 13 (15.38%) fresh semen samples collected from Pasuruan also showed to be positive. In addition, the nPCR was faster and easier to perform than the standard viral isolation test. It is concluded that, the nPCR can be used as test of choice for routine diagnosis of BHV-1. Key Words: Nested PCR, BHV-1, Semen, Glycoprotein D Gene, TCID50
Protective antibody titre against Newcastle disease in ostriches (Struthio camelus) ., Darminto; Bahri, Sjamsul; Suryana, N.
Indonesian Journal of Animal and Veterinary Sciences Vol 3, No 4 (1998)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (160.691 KB) | DOI: 10.14334/jitv.v3i4.124

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The aim of this study was to define an estimated antibody titre which was considered to be protective against Newcastle disease (ND) virus infection in ostriches. Eighteen young ostriches of 4 days of age were divided into two groups each containing 9 birds. The first group was unvaccinated and the second group was vaccinated against ND virus twice at 4 and 14 days of age. Antibody titres were monitored at 1, 14, 28, 42, 56, 70 and 85 days of age by haemagglutination inhibition (HI) test. All birds were then challenged with a velogenic strain of ND virus, Ita strain, at 42 days of age. The excretion of the challenge virus were monitored daily after challenge up to the end of this experiment. Several organs such as brain, trachea, lungs and spleen were collected from died birds for re-isolation of the challenged virus. Results indicated that all unvaccinated birds succumbed to the challenged virus, except one bird that survived challenged. In contrast to the unvaccinated birds, all vaccinated birds survived challenged, except two birds with low antibody titres succumbed challenged. All birds with antibody titres of 4 (HI-log2) or greater survived challenged. All challenged birds excreted the challenged virus through out their oropharyngs. Moreover, challenged virus can be successfully re-isolated from most organs of the died birds. This study concludes that : (a) the estimated protective titre against ND in ostriches is 4 (HI-log2), (b) the immune status for ostrich with antibody titre less the 4 (HI-log2) could not be defined, and (c) vaccination against Newcastle disease in ostriches could successfully prevent birds from sick and died of ND, but unable to prevent virus infection and unable to stop carrier status after infection. Key words : Newcastle disease, ostrich, antibody, protective titre
Development of inactivated-local isolate vaccine for infectious bronchitis ., Darminto
Indonesian Journal of Animal and Veterinary Sciences Vol 4, No 2 (1999)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (174.356 KB) | DOI: 10.14334/jitv.v4i2.147

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Infectious bronchitis (IB) is an acute highly contagious viral respiratory disease of poultry caused by coronavirus. The disease causes high mortality in young chicks, reduce body weight gain in broilers and remarkable drop in egg production. IB can only be controlled by vaccination, but due to the antigenic variation among serotypes of IB viruses, the effective IB vaccine should be prepared from local isolates. The aim of this research is to develop inactivated IB vaccine derived from local IB isolates. Local isolates of IB viruses designated as I-37, I-269 and PTS-III were propagated respectively in specific pathogen free (SPF) chicken eggs, the viruses then were inactivated by formaline at final concentration of 1:1,000. Subsequently, the inactivated viruses were mixed and emulsified in oil emulsion adjuvant with sorbitant mono-oleic as an emulsifier. The vaccine then was tested for its safety, potency and efficacy in broiler chickens. Birds inoculated twice with a two-week interval by inactivated vaccine did not show any adverse reaction, either systemic or local reaction. The inoculated birds developed antibody responses with high titre, while antibody of the control birds remain negative. In addition, efficacy test which was conducted in broilers demonstrated that birds vaccinated by live-commercial vaccine and boosted three weeks later by Balitvet inactivated vaccine showed high level of antibody production which provided high level of protection against challenged virus (76% against I-37, 92% against I-269 and 68% against PTS-III challenge viruses). From this study, it can be concluded that inactivated local IB vaccine is considered to be safe, potent and efficacious. The vaccine stimulates high titre of antibody responses, which provide high level of protection against challenged viruses.   Key words: Infectious bronchitis, virus, broiler chicken, antibody, inactivated vaccine
Seroepidemiologi Nipah Virus pada Kalong dan Ternak Babi di Beberapa Wilayah di Indonesia Sendow, Indrawati; Field, Hume; Adjid, R.M. Abdul; Syafriati, Tatty; Darminto, Darminto; Morrissy, Chris; Daniels, Peter
JURNAL BIOLOGI INDONESIA Vol 5, No 1 (2008): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (146.164 KB) | DOI: 10.14203/jbi.v5i1.3205

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ABSTRACTNipah Virus Seroepidemiology in Flying Fox and Pig Husbandry in Several Areasof Indonesia. Nipah is a dangerous zoonotic disease which was carried by flying fox.The disease had been occurred in Malaysia in 1999 and infect pigs and caused humandeath. Indonesia is adjacent country to Malaysia, hence, a serological study had beenconducted on 156 flying fox (P. vampyrus) sera from North Sumatera, West Java, CentralJava and East Java. Besides that, 2740 pig sera was randomly collected in differentprovinces to detect Nipah infection. Both flying fox and pig sera were tested usingELISA test to detect the presence of Nipah antibody. The results indicated that 37 from156 flying fox sera (23.7%) has antibodies against Nipah virus. Infections were occuredin all sampling sites with the prevalence varied from 18% to 33 %. Meanwhile, no pigsera tested (2740) had antibody against Nipah virus. Based on these results it can beconcluded that Nipah virus infections were occurred in flying fox in some parts inIndonesia, but not in pigs. It was suggested that the presence of Nipah virus in Indonesiashould be anticipated. Hence the distribution of its infection in pigs and human must beanticipated. Monitoring of Nipah infection in areas adjacent to Malaysia must be increasedto detect the entering of the disease in Indonesia.Keywords: Nipah, pigs, flying fox, serology
Pemetaan Genetik Virus Avian Influenza di Indonesia 2007 Dharmayanti, NLP Indi; Indriani, R.; Hartawan, R.; Hewajuli, D.A.; Ratnawati, A.; Darminto, Darminto
JURNAL BIOLOGI INDONESIA Vol 5, No 2 (2008): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (667.831 KB) | DOI: 10.14203/jbi.v5i2.3195

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ABSTRACTGenetic Mapping of Indonesian Avian Influenza Viruses 2007.Department of agricultureimplements vaccination as one a tool to control of avian influenza disease. The vaccinationprogram use virus seed such as H5N1, H5N2 and H5N9. Such as vaccination program for otherdiseases, avian influenza vaccine program have lack implementation in the field. In 2007,department of agriculture evaluated the AI vaccination program such as the master seedvaccine that can be used. Result of the evaluation showed that 11 of AI vaccines that weredistributed in Indonesia did not give protection more than 60% to Indonesian isolates in 2006(A/Ck/Pwt-Wij/06). From this point and many AI cases in the field in 2007, the aim of thisstudy was to conduct genetic diversity of avian influenza viruses which have circulated inIndonesia region. We used virus isolation for propagate the viruses, RT-PCR for identificationand DNA sequencing on HA1 region to analysis genetic diversity for genetic mapping anduseful for master seed candidate. The result of the study showed that there were 6 group ofgenetic diversity in 2007; Isolates from group 1, 5 and 6 can be used for AI vaccine candidate.Key words: genetic mapping, genetic diversity and avian influenza viruses
Co-Authors A Ratnawati A Wiyono Abdullah Shahab abdurrouf Abdurrouf Adler Haymans Manurung Agung Suwandaru Agus Wiyono Ahzan, Sukainil Alimuddin Ali Anne Zulfia Arifin, Rizal Bambang Subali Budi Priyanto Cahyoingrat, Suryo E. Cahyono, Yoyok D.A Hewajuli Dadan Hamdani, Dadan Daniels, Peter Daniels, Peter Dedi Supriadi Dharmayanti, N.L P.I. Dharmayanti, N.L P.I. Dharmayanti, NLP Dharmayanti, NLP Eddy S Siradj Eko Handoyo Endang Siti Astuti Endhah Purwandari Fahmi Astuti, Fahmi Fatimah Karunia Sultera Field, Hume Field, Hume Firdaus, Aulia Anisa Gatut Yudoyono H Hamid Hartawan, R. Hartawan, R. Helmy Hamid Hesti Irnanta Hewajuli, D.A. Hewajuli, D.A. I Wayan T. Wibawan Ida Ayu Putu Sri Widnyani Ida Widaningrum, Ida Indra Sidharta Indrawati Sendow Intifadhah, Sahara Hamas Iwan Dini Iwan Yahya Jagoda Ignjatovic Johari, Norhasnidawani Joni Dwi Pribadi Kholis Nurhanafi Kolif Yulianto L Parede Lee, Vannajan Sanghiran Lintang Edityastono Manik, Tetti Novalina Martin Malole Mashuri Mashuri Morrissy, Chris Morrissy, Chris Muhammad Taqiyuddin Muhammad Zainuri Muharam Saepulloh Munir, Rahmawati Muzakki, Fikrun Najib N. Suryana N.L.P Indi Dharmayanti N.L.P.I Dhamaryanti N.L.P.I Dharmayanti N.L.P.I. Dharmayanti Ninis Hadi Haryanti, Ninis Hadi NLP Indi Dharmayanti Nugroho, Ferry Anggoro Ardy Nuni Widiarti Nurma Sari, Nurma Nurshabrina, Atiqa Taqiyya P Ronohardjo Prayogi, Saiful Prayogi, Soni Puspita, Niniek Fajar R Damayanti R Indriani R M A Adjid R. Indriani R.M Abdul Adjid R.M. Abdul Adjid R.M.A. Adjid Ratnawati, A. Ratnawati, A. Retno Asih, Retno RISA INDRIANI Risa Indryani Risdiana, Risdiana Rizoputra, Ivan Rizqy Wahyu Septiono S Hardjoutomo Saleh, Muh Santoso, Tanto Budi Sembiring, Rinawati Shaliha, Shaliha Shinta Maharani Trivena Sholih, Ahmad Simon Sadok Siregar, Simon Sadok Simson Tarigan Siti Ragil Handayani Sjamsul Bahri SRI WARDANI Suarso, Eka Suhadak Suhadak Suminar Pratapa Suryajaya Suryajaya, Suryajaya Susan M Noor Susi Soviana Sutikno Sutikno Syah, Rakhasoni Firman T Sjafriati T Syafriati Taher Al habsji Tatty Syafriati Totok Wianto Upik Kesumawati Hadi W. Asmara W. T. Artama, W. T. Wahyudi, Sriati Winardi, Yoyok Zainul Arifin Zainuri, M Zulkarnain, Zulkarnain