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Immunological Detection of Rabies Virus in Brain Tissues of Infected Dogs by Monoclonal Antibodies Nyoman Mantik Astawa; Ida Bagus Kade Suardana; Luh Putu Agustini; Faiziah -
Jurnal Veteriner Vol 11 No 4 (2010)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

In order to establish an immunological detection of rabies virus in tissues of infected dogs, monoclonalantibodies (mAbs) against rabies virus (RV) were produced. The mAbs were produced by fusion of mielomacells with the lymphocytes of mice immunized with RV. The mAbs produced were then characterized andused for the detection of rabies virus in brain tissues of infected dogs. Six mAbs designated CC6, EG4,DG10, BB12, CA9 dan EB5 were used in this study. In Western blotting test, some mAbs reacted with 66KDa which is the glycoprotein of the virus. In immunoperoxidase, 2 mAbs (CC6 and DG10) detected RVin the brain of infected dogs. By direct immunoflourescence, flourescence isotyocyanate (FITC) labelledDG10 mAbs detected RV in fresh and formaldehyde fixed brain tissues. RV was detected in 12 infecteddogs but not in normal uninfected dogs. In this study it was confirmed that rabies virus can be detected inthe brain tissues of infected dogs by monoclonal antibodies.
Karakteristik Molekuler Virus Avian Orthoavulavirus 1 Genotipe VII yang Diisolasi dari Tabanan Bali (MOLECULAR CHARACTERISTIC OF AVIAN ORTHOAVULAVIRUS 1 GENOTYPE VII ISOLATED FROM TABANAN BALI) Anak Agung Ayu Mirah Adi; I Nyoman Mantik Astawa; I Nengah Wandia; I Gusti Agung Arta Putra; Ida Bagus Oka Winaya; Anak Agung Keswari Krisnandika; Anak Agung Oka Wijaya
Jurnal Veteriner Vol 20 No 4 (2019)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Newcastle disease (ND) is a very harmful avian disease, endemic in Indonesia and various parts of the world. The causative agent is ND virus or Avian orthoavulavirus 1 (AOAV-1). This virus is an RNA virus with wide genetic variation. Based on the genome length, it can be classified into AOAV-1 Class I and II. Class I are generally avirulent whereas Class II are consist of both virulent and avirulent viruses, currently there are 18 genotypes of the class II. To find out the molecular characteristics of AOAV-1 currently circulating in the field, isolation and identification of viruses from laying hens that was suspected ND from Tabanan Bali in 2017, was performed. The isolated viruses hereafter named as Tabanan1/ARP / 2017. A one-step RT-PCR reaction was carried out to amplify NP, F and HN gene fragments from the virus using three specific pairs of AOAV-1 primers. The obtained nucleotide sequences are then used in phylogenetic analysis. For phylogenetic analysis several strains of AOAV-1 from class II representing genotype I-VII as well as one strain from Class I were accessed from GenBank. From the analysis of the F gene nucleotide sequences, it was found that Tabanan 1 / ARP / 2017 is a genotype VII virus with an amino acid sequence at the F protein cleavage site is 112 R-R-Q-K-R-F117, a typical virulent strain. Phylogenetic analysis using nucleotide sequences NP and HN genes also positioned this isolate in genotype VII. At the nucleotide level, genetic distance with virulent isolates that was isolated in 2007 and 2010 were 8.26% and 1.08% while at the amino acid level were 5.26% and 0.64%. There were found mutations in amino acids at positions 107 and 108 of F protein.
REACTIVITY OF ANTI-CAPSID MONOCLONAL ANTIBODIES WITH NATIVE AD RECOMBINANT PROTEIN OF JEMBRANA DISEASE VIRUS REAKTIVITAS ANTIBODI MONOKLONAL ANTI-KAPSID DENGAN PROTEIN NATIF DAN PROTEIN REKOMBINAN VIRUS PENYAKIT JEMBRANA Nyoman Mantik Astawa
Jurnal Veteriner Vol 8 No 2 (2007)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Reactivity of anti-capsid monoclonal antibody (mAbs) with both native and recombinant protein of Jembrana Disease Virus (JDV) was examined. The monocloncal antibodies were produced by fusion of my.loma cells with lymphocytes of mice immunized with JDV antigen. Ten mAbs were produced and they designated as CC12, BB7, AH7, BD2, DB2, AF9, BC10, AG7, AH4 and CB11. In ELISA test, all mAbs reacted with recombinant proteins (glutation-8-transferase-capsid/GST-Ca and histidine tagged capsid/His-Ca) but not with GST alone.
Kloning, Sikuensing dan Analisis Filogenetik Gen Nukleokapsid Protein Virus Tetelo Isolat Bali-1/07 Anak Agung Ayu Mirah Adi; I Made Kardena; Nyoman Mantik Astawa; Ketut Santhia Adhy Putra; Yoshihiro Hayashi; Yasunobu Matsumoto
Jurnal Veteriner Vol 12, No 3 (2011)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

A study was carried out on the molecular characteristics of gene encoding for nucleocapsid protein(NP) of Newcastle disease virus (NDV) Bali-1/07. A portion of the fragment gene was amplified by reversetranscriptase-polymerase chain reaction (RT-PCR). The RT-PCR product purified from the gel was treatedwith Taq polymerase and cloned into plasmid pT7blueT vector. The recombinant plasmid was tranfectedinto Nova blue singles strain of Escherichia coli-competent cells and selected using white and blue colorselection method. Good white colonies were subcultured and tested for presence of expected gene fragmentby polymerase chain reaction (PCR). The correct size PCR product was recloned and sequenced. The PCRproduct of 540 base pairs were sequenced and aligned with several cognates genes of world NDVisolates.using Cluster IW. The phylogenetic analysis was then performed using MEGA 4. Phylogeneticanalysis showed that the NP gene of NDV Bali-1/07 is closely related with virulent NDVs such asGuangxii-11/03, KBNP-4152 and Ast2755/01 with nucleotide sequence homology of 92%, 91.4% and91%respectively, whereas the nucleotide sequence homology with avirulent NDVs such as LaSota and B1were 77%.
Lymphocytes Subpopulation in Peripheral Blood and Spleen of Village Chickens Recognized by Monoclonal Antibodies (SUBPOPULASI LIMFOSIT PADA DARAH TEPI DAN LIMPA AYAM KAMPUNG YANG DIKENALI OLEH ANTIBODI MONOCLONAL) Nyoman Mantik Astawa; Ida Bagus Made Oka
Jurnal Veteriner Vol 17 No 2 (2016)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Lymphocytes play important role in host defence system against pathogenic agents both in mammalianand avian species. Monoclonal antibodies (mAbs) have been widely used to identify lymphocytessubpopulation in a host based on their surface cluster differentiation (CD) markers. Currently, mAbsagainst lymphocytes surface markers of village chickens have been produced by fusion of myeloma withlymphocytes derived from spleen of mice immune to chicken lymphocytes. In two fusion experiments, 623clones of hybridomas were produced and four (BG4, CB1, DB2 and BB2) of which secreted mAbs againstchickens lymphocyte surface molecules. Two mAbs (BG4 and DB2) recognized protein of 32 kDa, one mAb(CB1) recognized protein of 64 kDa, and one mAb was unable to recognize any protein of chicken lymphocytesurface molecule. Three mAbs recognized lymphocyte subpopulation in spleen and peripheral blood ofvillage chickens. In peripheral blood, mAbs BG4, CB1 and DB2 recognized lymphocytes subpopulationwith the percentages of 11.2%, 21.4% and 7.4% respectively. In spleen those three mAbs recognizedlymphocytes subpopulations at the percentages of 38.2%, 51.54% and 31.5% respectively. Based on thoseresult, it is very likely that mAbs BG4 and DB2 recognized CD4 molecule and mAb CB1 recognized CD8molecule of village chickens lymphocytes.
Pelacakan Secara Imunohistokimiawi Antigen Virus pada Ayam yang Diinfeksi dengan Virus Penyakit Tetelo (IMMUNOHISTOCHEMICAL DETECTION OF VIRAL ANTIGEN IN TISSUE OF CHICKENS EXPERIMENTALLY INFECTED WITH NEWCASTLE DISEASE VIRUS) Anak Agung Ayu Mirah Adi; I Made Kardena; Nyoman Mantik Astawa; Yasunobu Matsumoto
Jurnal Veteriner Vol 13 No 3 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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In order to study the distribution of Newcastle disease virus (NDV) following infection, chickenswere experimentally infected with visceretropic velogenic NDV isolate. Monoclonal antibodies (mAbs)against the NDV LaSota vaccine strain were then produced to detect viral antigen in the infectedorgans. The mAbs were firstly tested for their specificity by enzyme linked immunosorbent assay(ELISA) using NDV and normal allantoic fluids as antigens. Eight mAbs specific against NDVwere isolated and two mAbs were used for immunodetection of NDV antigen in chicken’s tissues.By immunohistochemistry labeled streptavidin-biotin (LSAB) staining NDV–antigen was detectedin paraffin embedded tissues of NDV-infected chickens. NDV antigen was not detected in noninfected chickens. In the infected chickens, high intensity of NDV antigen was detected in thelymphoid tissues, lung and intestine. The NDV antigen with a lesser intensity was detected in thebrain, trachea, liver and myocardium. This study shows that although viscerotropic velogenicNDV isolate can infect almost all organs, the main target of infection are lung, intestine andlymphoids tissues
Mosquito-specific viruses (family Flaviviridae, genus Flavivirus) Diisolasi pada Nyamuk Anopheles vagus di Bali Putu Ayu Asri Damayanti; I Nyoman Mantik Astawa; Anak Agung Ayu Mirah Adi; I Made Sudarmaja; I Kadek Swastika; Dewa Ayu Agus Sri Laksemi; Ni Luh Putu Eka Diarthini
Jurnal Veteriner Vol 22 No 2 (2021)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Mosquito-specific viruses (MSVs) adalah virus yang hanya dapat bereplikasi pada sel nyamuk. Virus ini terdiri dari berbagai genus, salah satunya yang paling banyak ditemukan adalah dari famili Flaviviridae, genus Flavivirus. Namun, data keberadaan dan karakteristik MSVs dan vektornya di Bali saat ini sangat terbatas. Oleh karena itu, pengamatan untuk memperluas penemuan keragaman vektor dan filogenetik MSVs famili Flaviviridae, genus Flavivirus di Bali dilakukan pada tahun 2016-2018. Nyamuk dewasa ditangkap menggunakan light trap dan dikelompokkan berdasarkan spesies. Isolasi dan propagasi virus dilakukan pada galur sel C6/36 dan baby hamster kidney-21 (BHK-21). Identifikasi virus dilakukan dengan menggunakan one step reverse-transcriptase polymerase chain reaction (RT-PCR). Terdapat dua pool yang berasal dari nyamuk Anopheles vagus menampakan cythopathic effect (CPE) hanya pada galur sel C6/36 dari total 158 pool. Virus yang diisolasi memiliki persentase identity sekuen nukleotida tertinggi 97% dan sekuen asam amino 96% dengan virus Culex theileri Flavivirus isolat JKT-8650 yang diisolasi pada tahun 1981. Selanjutnya, virus dinamakan Mosquito Flavivirus Isolate Bali (MFB) dengan accession numbers KY995166 dan KY290258. Analisis filogenetik menunjukan bahwa MFB berada satu kluster dengan Culex theileri Flavivirus (CTFV) dari Indonesia, Culex Flavivuruses-Myanmar, Culex theileri Flavivirus-Portugal, dan Mosquito Flavivirus-Turki. Terdapat delapan nukelotida dan enam asam amino yang berbeda antara MFB dan CTFV Indonesia. Pada penelitian ini dapat disimpulkan bahwa MSVs dari famili Flaviviridae, genus Flavivirus berhasil diisolasi dari nyamuk An. vagus di Bali.
Ekstrak Pegagan Meningkatkan Titer Antibodi Mencit Setelah Diinfeksi Salmonella typhi (CENTELLA ASIATICA EXTRACT INCREASE ANTIBODY TITER IN MICE AFTER SALMONELLA TYPHI INFECTION) I Nengah Kerta Besung; I Nyoman Mantik Astawa; I Ketut Suatha; Hartaningsih .
Jurnal Veteriner Vol 14 No 2 (2013)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

A study was conducted to find out the ability of Centella asiatica (C. asiatica) in enhancing antibodyresponse of C. asiatica treated mice following Salmonella typhi (S. typhi) infections. It is therefore expectedthat herbal drug such as  C. asiatica  can be used as an alternative medicine to prevent and to curesalmonellosis both in animals and human. Experimental laboratory studies were conducted usingCompletely Factorial Randomized Design. Mice were divided into four groups and they were treatedrespectively with destilated water (negative control), 125, 250, and 500 mg/kg BW/day of  C. asiaticaextract. The treatment was conducted daily for two weeks  and the mice were inoculated with 105 cells/mlof  S. typhi. The antibody response were examined by indirect enzyme-linked immunosorbent assay (ELISA)on first day, second week and fourth week  after S. typhi infections.  The result showed that treatment ofmice with C. asiatica extract significantly (p<0,05) enhanced antibody titer of Balb/c mice after S. typhiinfections. The highest antibody titer was observed at four weeks after S. typhi infections with 500 mg/kgBW/day (94,0370 ± 1,69 IU).
Seroprevalensi Sistiserkosis pada Babi di Papua (SEROPREVALENCE OF PIG CYSTICERCOSIS IN PAPUA REGION) Ida Bagus Ngurah Swacita; I Ketut Suada; Ketut Budiasa; Nyoman Sadra Dharmawan; Nyoman Mantik Astawa; I Nyoman Polos; I Made Damriyasa
Jurnal Veteriner Vol 18 No 1 (2017)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Pig cysticercosis is an infection caused by the larval stage of pork tapeworm and Papua is one of the largest endemic areas of cysticercosis in Indonesia. This survey aim was to determine the seroprevalence of pig cystisercosis in Papua. A total of 311 pig serum samples collected from six regencies in Papua were examined using Enzyme Linked Immunosorbent Assay (ELISA). The result of the survey showed that the average seroprevalence of pig cysticercosis in Papua was 23.5% (73/311), where the highest seroprevalence was found in the regency of Jayawijaya was 42.6% (43/101), Biak 22.5% (9/40), Nabire 20.6% (7/34), Mimika 17% (8/47), Jayapura 13.5% (5/37), and Merauke 1.9% (1/52). It can be concluded that the seroprevalence of pig cysticercosis in Papua is still high, therefore, it is necessary to do more intensive programs to prevent and control this disease. ABSTRAK Sistiserkosis pada babi adalah infeksi yang disebabkan oleh stadium larva cacing pita, dan Papua merupakan salah satu daerah endemis sistiserkosis di Indonesia. Tujuan penelitian ini adalah untuk menentukan seroprevalensi sistiserkosis pada babi di Papua. Sebanyak 311 sampel serum babi yang dikumpulkan dari enam kabupaten di Papua diuji dengan Enzyme Linked Immunosorbent Assay (ELISA). Hasil penelitian menunjukkan bahwa rataan seroprevalensi sistiserkosis pada babi di Papua sebesar 23,5% (73/311), dan seroprevalensi terbesar ditemukan di Kabupaten Jayawijaya 42,6% (43/101), Biak 22,5% (9/40), Nabire 20,6% (7/34), Mimika 17% (8/47), Jayapura 13,5% (5/37), dan Merauke 1,9% (1/52). Dapat disimpulkan bahwa seroprevalensi sistiserkosis pada babi di Papua masih tinggi, sehingga diperlukan program yang lebih intensif untuk mencegah dan mengontrol penyakit ini.
Respons Antibodi Sekunder Terhadap Penyakit Tetelo pada Ayam Petelur Pascavaksinasi Ulangan dengan Vaksin Tetelo Aktif (NEWCASTLE DISEASESECONDARY ANTIBODY RESPONSE AFTER REVACCINATION IN LAYER WITH THE ACTIVE ND VACCINE) Andika Budi Kurnianto; Gusti Ayu Yuniati Kencana; I Nyoman Mantik Astawa
Jurnal Veteriner Vol 17 No 3 (2016)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Revaccination is required in order to preventNewcastle Disease (ND) reccurence inlayers chickens. Oneof vaccine for ND revaccination is freeze-died ND active vaccine containing e” 106,5EID50. Revaccinationisdone to trigger a faster secondary antibody responses in layers and can achieve protective antibody titersagainst ND that can be monitored by a hemagglutinationinhibition (HI). The aim of this study was todetermine the ND secondary antibody responses in layers after revaccination with ND active vaccine.Antibody titer of 20 layers chickens of 20 week old were determined before revaccinations (week 0) andafter revaccinations (week 1 until week 9). The first vaccination was conducted using ND-IB (NewcastleDisease-Infectious Bronchitis) at the age of 2 days through eye drops and subcutaneous injection at the ageof 5 days using a dose of 1 ampoule.Vaccination is repeated at the age of 20 weeks at a dose of 1 ½ ampoule through drinking water. Blood samples were collected on the wing vein (venous cutane ulnar) and left for 5-10 minutes at room temperature.Sera were then collected and stored at -20oC until use. HI antibody titerwas determined by micro titeration system. The HI mean titers were analyzed by Duncan test. The studyresults showed that antibody titer before revaccination was3,47 HI log 2 and the HI titers after revaccinationwere 4,02; 5,22; 6,52; 7,85; 8,4; 8,6; 7,7; 5,92; dan 3,87 HI log 2 respectivelly at weeks 1, 2, 3, 4, 5, 6, 7, 8, and9.The NDV revaccination with ND active vaccine significantly (P <0.01) increased in antibody titer inlayers starting from week 1 to week 6, but decreased following week 7 to week-9. It can be concluded thatrevaccinantion with ND active vaccine increases the antibody titers in layer chickens.
Co-Authors A. A. G. P. Wiraguna A.A. Wiradewi Lestari A.A.G. Sudewa AAG Budhitresna AAG Putra Ahmad Hamim Sadewa Aida Lousie Tenden Rompis Alberto Agustinho Pereira Da Costa Joao Anak Agung Ayu Mirah Adi Anak Agung Bagus Bramardipa Anak Agung Gede Sudewa Djelantik Anak Agung Keswari Krisnandika Anak Agung Ngurah Subawa Anak Agung Oka Wijaya Anak Agung Sagung Kendran and R. Kusnandi Andika Budi Kurnianto Anwar Santoso Arthawan Arthawan Bayu Setiabudi Berata , I Ketut Chandra Yowani Dewa Ayu Agus Sri Laksmi DWI SURYANTO Dyah Kanya Wati Faiziah - G.A.M.K. Dewi Gusti Ayu Mayani Kristina Dewi Gusti Ayu Yuniati Kencana Gusti Ngurah Narendra Putra Harjana, Ngakan Putu Anom HARTANINGSIH - Hartaningsih . I D. N. Wibawa I Dewa Made Sukrama I Gede Ngurah Harry Wijaya Surya I GEDE PUTU WIRAWAN I Gusti Agung Arta Putra I Gusti Agung Ayu Suartini I Gusti Agung Dewi Sarihati I Gusti Agung Trisna Windiani I Gusti Ayu Putu Eka Pratiwi I Gusti Made Krisna Erawan I Gusti Ngurah Kade Mahardika I Gusti Ngurah Sudisma I K. Sukardika I Kadek Swastika I Ketut Berata I Ketut Eli Supartika I Ketut Junitha I Ketut Suada I Ketut Suada I Ketut Suastika I Ketut Suastika I KETUT SUATA I Ketut Suatha I Ketut Suwiyoga I Made Bakta I Made Damriyasa I Made Dwinata I Made Galih Diparayoga I Made Jawi I Made Kardena I Made Subrata I Made Sudarmaja I Made Sukada i Nengah Wandia I Nyoman Agus Bagiada I Nyoman Budi Hartawan I Nyoman Polos I Nyoman Suarsana I Nyoman Suartha I Putu Sudiarta I W. Wita, I W. I Wayan Bebas I Wayan Gorda I Wayan Masa Tenaya I Wayan Megadhana I Wayan Putu Sutirta Yasa I Wayan Suardana I Wayan Wita I.A.P. Apsari I.B.K. Suardana I.H. Utama I.W. Batan Ida Ayu Pasti Apsari Ida Bagus Gede Suparyatha Ida Bagus Kade Suardana Ida Bagus Made Oka Ida Bagus Ngurah Swacita Ida Bagus Oka Winaya Ida Bagus Suardana Ida Bagus Subanada Ignatius Ferdi Yuatmadja Inna Narayani K. Sri Marhaeni Julyasih K. Suata K. Sukardika Kadek Karang Agustina Ketut Budiasa Ketut Santhia Adhy Putra Ketut Suata Ketut Tirtayasa Luh Dewi Anggreni Luh Putu Agustini LUH PUTU AGUSTINI Luh Putu Wrasiati M.D. Rudyanto M.Pd S.T. S.Pd. I Gde Wawan Sudatha . Made Damriyasa, Made Made Oka Ari kamayani, Made Oka Ari Made Wiryana Marissa Divia Dayanti Marson, Fransiska Gratia Sonita Mudinillah, Adam N. T. Suryadhi Ngakan Putu Anom Harjana Ni Luh Putu Eka Diarthini NI LUH PUTU MANIK WIDIYANTI Ni Made Krisna Dewi Ni Made Suaniti NINING HARTANINGSIH Nyoman Agus Bagiada Nyoman Tigeh Suryadi Oka Lely Palagan Senopati Sewoyo Purwitasari, Made Santi Putu Ayu Asri Damayanti Putu Ayu Sisyawati Putriningsih Rasmaya Niruri Rasmaya Niruri S. Soetjiningsih, S. S. Sotjiningsih S.K. Widyastuti sang gede purnama Sewoyo, Palagan Senopati Siti Maryam Soetjiningsih - Soetjiningsih . Sri Kayati Widyastuti Sri Wahjuni SUMARNO Suryadi, Nyoman Tigeh Sutjahjo Suherman, Sutjahjo Suwarno - T. Sari Nindia Tjok Gede Oka Pemayun, Tjok Gede Oka Tjokorda Gde Agung Suwardewa Wayan Tunas Artama Wimpie I Pangkahila Yasunobu Matsumoto Yosevangelika Hutabarat Yoshihiro Hayashi