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Penyebaran Virus Vaksin ND Pada Sekelompok Ayam Pedaging Yang Tidak Divaksinasi dan dipelihara bersama ayam yang divaksinasi Kencana, Gusti Ayu Yuniati; Astawa, Nyoman Mantik; Mahardika, I Gusti Ngurah Kade; Gorda, I Wayan
Buletin Veteriner Udayana Vol. 4 No.2 Agustus 2012
Publisher : Buletin Veteriner Udayana

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Abstract

Telah dilakukan penelitian untuk mengetahui daya sebar vaksin ND aktif galurlentogenik (La Sota) dan respons immune ayam yang tidak divaksin yang dipeliharabersama ayam yang divaksin secara intramuskuler. Penelitian ini menggunakan rancanganacak lengkap pola berjenjang (split time) dengan faktor utama perlakukan vaksinasi (TO:0% divaksin dan 100% tidak divaksin , T1: divaksin 50 % dan 50 tidak divaksin dan T2:divaksin 75% dan 25% tidak divaksin) dengan sembilan kali ulangan. Faktor tambahanadalah waktu pengambilan serum (minggu ke-0, ke-1, ke-2 dan ke-3) sehingga jumlahsampel adalah 3x9x4= 108 sampel serum. Ayam umur 3 hari divaksinasi ND secara tetesmata kemudian dilakukan vaksinasi intramuskuler pada umur 21 hari sesuai perlakuan.Titer antibodi ND pada ayam perlakuan diuji dengan uji hambatanhemaglutinasi/hemagglutination inhibition (HI) satu hari sebelum vaksinasi, serta satuminggu, dua minggu, dan tiga minggu setelah vaksinasi. Data tentang titer antibodi (GMTHI)terhadap ND ditransformasi dengan akar X+1, dianalisis dengan sidik ragam dandilanjutkan dengan uji jarak berganda Duncan. Hasil penelitian menunjukkan bahwa titerantibodi terhadap ND pada ayam yang tidak divaksin dipengaruhi oleh persentase ayamyang divaksin. Antibodi HI unit terhadap virus ND pada ayam yang tidak divaksinasimulai teramati pada minggu ke-2 dan ke-3 setelah vaksinasi. Titer antibodi ayam yangtidak divaksinasi pada kelompok ayam yang hanya divaksin 75% mempunyai titer antibodiyang nyata lebih tinggi dibandingkan dengan kelompok ayam yang divaksin 50% dankontrol (P<0,05). Pada kelompok ayam yang divaksin 50%, titer antibody ND pada ayamyang tidak divaksin secara statistik berbeda tidak nyata dibandingkan dengan kelompokyang divaksin 0% (P>0,05). Pada minggu ke tiga, titer antibody ND ayam yang tidakdivaksinasi pada kelompok ayam yang divaksin 75% nyata lebih tinggi dibandingkandengan pada kelompok ayam yang divaksin 50% (P,0,05). Vaksin ND aktif lentogeik LaSota dapat menyebar dari ayam yang divaksin secara intramuskuler kea yam yang tidakdivaksin
Immunological Detection of Avian Influenza Virus in Infected Ducks by Monoclonal Antibodies Against AIV-H5N1 ASTAWA, NYOMAN MANTIK; WINAYA, IDA BAGUS OKA; AGUSTINI, LUH PUTU; HARTANINGSIH, NINING
Microbiology Indonesia Vol 1, No 3 (2007): December 2007
Publisher : Indonesian Society for microbiology

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Abstract

In order to establish a detection method for avian influenza virus (AIV) infection in ducks, monoclonal antibodies (MAbs) against the virus were produced. The virus used for the production of the monoclonal antibodies was AIV-H5N1 of Indonesian origin. Immortal mouse myeloma were fused with the lymphocytes derived from the spleen of mice immunized with the virus. The MAbs were tested for their specificity by enzyme linked immunosorbent assay (ELISA) and western blotting using formaldehyde inactivated virus and normal allantoic fluid as a negative control. Twelve MAbs which were specific against AIV were isolated and 8 of them were used for detecting of AIV antigen in duck’s tissues. AIV antigen was detected in paraffin embedded tissues of AIV-infected ducks by immunohistochemistry using MAbs. AIV antigen was not detected in ducks, which were confirmed to be AIV negative. In the infected ducks, high intensity of AIV infection was detected in proventricle gland and small intestine. The AIV antigen with a lesser intensity was also detected in lungs, spleen, and bursa of Fabricius, but hardly detected in muscle, brain, and several other issues. This study shows a clear evidence that MAbs produced in this study are applicable for use in immunological detection of AIV in infected duck tissues.
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) Mirah Adi, Anak Agung Ayu; Kardena, I Made; Astawa, Nyoman Mantik; Matsumoto, Yasunobu
Jurnal Veteriner Vol 13, No 3 (2012)
Publisher : Jurnal Veteriner

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Abstract

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
The Production and Use of Monoclonal Antibodies for the Detection of Avian Influenza Antigen in the in Infected Chickens Astawa, Nyoman Mantik; Suardana, Ida Bagus Kade; Kencana, Gusti Ayu Yuniati
Jurnal Veteriner Vol 13, No 3 (2012)
Publisher : Jurnal Veteriner

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Abstract

A safe and appropriate diagnostic method for avian influenza virus (AIV) infection in chickens wasestablished using monoclonal antibodies (mAbs) against the virus. The virus used for the production of themonoclonal antibodies was an Indonesian AIV-H5N1 isolate. Immortal mouse myeloma cells were fusedwith the lymphocytes derived from the spleen of mice immunized with the virus. The mAbs were tested fortheir specificity by enzyme linked immunosorbent assay (ELISA) and western blotting using formaldehydeinactivated virus and normal allantoic fluid as antigens. Twelve mAbs specific against AIV were isolatedand 8 mAbs were used for immunodetection of AIV antigen in chicken’s tissues. By indirect ELISA, themAbs were able to detect AIV antigen in allantoic fluid at the titre as low as 2-2 to 2-4 HA units per 0.1 ml.By immunoperoxidase staining AIV–antigen was detected in paraffin embedded tissues of AIV-infectedchickens. AIV antigen was not detected in chickens which were confirmed to be AIV negative. In theinfected chickens, high intensity of AIV antigen was detected in proventricle gland and small intestine.The AIV antigen with a lesser intensity was detected in lungs and spleen but hardly detected in muscle,brain and several other tissues. This study show clear evidences that mAbs produced in this study areapplicable for use in the detection of AIV antigen in infected chickens.
Immunological Detection of Newcastle Disease Viral Antigen in the Naturally Infected Chickens by Monoclonal Antibodies against Fusion-2 Protein Astawa, Nyoman Mantik; Mirah Adi, Anak Agung Ayu
Jurnal Veteriner Vol 15, No 2 (2014)
Publisher : Jurnal Veteriner

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Abstract

Monoclonal antibodies (mAbs) against Fusion (F)2 protein of Newcastle disease virus (NDV) wereproduced for the detection of the viral antigen in infected chickens. Cells derived from spleen of Balb/c miceimmunized with the virus were fused with mouse myeloma cells to generate hybridomas capable ofproducing mAbs against the virus. The hybridomas were screened by enzyme-linked immunosorbent assay(ELISA) for anti-NDV specific mAbs using crude viral antigen (allantoic fluid of NDV-infected fertile eggs)and normal uninfected allantoic fluid of fertile eggs as negative control. The NDV proteins reactive withmAbs were then determined by Western Blotting using purified NDV as antigen. The mAbs reactive withF2 (12.5 KDa) protein of NDV were then used for the detection of NDV antigen in both the allantoic fluidof NDV- infected chicken embryos and in organs of naturally infected chickens. The results showed that 2out of 5 mAbs produced were against F2 protein of NDV. By indirect ELISA, the mAbs were able to detectthe viral antigen in allantoic fluid of NDV infected fertile chicken eggs at the titre as low as 2-2 to 2-4 HAunits per 0.1 mL. NDV–antigen was also detected by immunoperoxidase staining in paraffin-embeddedtissues of NDV-infected chickens but not in normal uninfected chickens. The most prominent infection wasdetected in the gastrointestinal tract and the lung. The NDV antigen was also detected in other organssuch as the brain, spleen, and several other tissues. It is evident that mAbs produced against F2 proteinof NDV were applicable for use in the detection of NDV antigen in infected chickens.
Respons Imun Mencit yang Diimunisasi dengan Cysticercus cellulosae (IMMUNE RESPONSE TO TAENIA SOLIUM CYSTICERCOSIS IN MICE) Swacita, Ida Bagus Ngurah; Damriyasa, I Made; Dharmawan, Nyoman Sadra; Astawa, Nyoman Mantik; Apsari, Ida Ayu Pasti; Tenaya, I Wayan Masa
Jurnal Veteriner Vol 16, No 2 (2015)
Publisher : Jurnal Veteriner

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Cysticercosis is a zoonotic parasitic disease which is still problem in Indonesia. The purpose of thisstudy was to investigate immune responses of mice that had been immunized using Taenia solium larval(Cysticercus cellulosae) antigens originated from infected pigs. Three kinds of the C.cellulosaeantigens,secretory and whole antigens were used to immunize three different groups Balb/c mice consisted of 15animals. The serum samples before and after immunization were tested with ELISA test. The resultsshowed that the third antigens induced highly significant titre (P<0,01)compared to unimmunized animals.However no significant different (P>0,05) were found when the third antigens were compared. It wasconcluded that immunization with the three kinds of C. cellulosae antigens can cause immunity in mice.
Produksi dan Karakterisasi Antibodi Monoklonal Anti-Cysticercus cellulosae (PRODUCTION AND CHRACTERIZATION OF MONOCLONAL ANTIBODIES AGAINST CYSTICERCUS CELLULOSAE) Swacita, Ida Bagus Ngurah; Damriyasa, I Made; Dharmawan, Nyoman Sadra; Astawa, Nyoman Mantik; Apsari, Ida Ayu Pasti; Oka, Ida Bagus Made; Tenaya, I Wayan Masa
Jurnal Veteriner Vol 16, No 3 (2015)
Publisher : Jurnal Veteriner

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Abstract

The purpose of this study is to make a monoclonal antibody against- Cysticercus cellulosae and itscharacterization. Samples antigen prepared from T. solium larvae (C. cellulosae) was then used to immunizeBalb/c. The immune response of mice assessed by ELISA test, then the lymphocytes of mice used for theproduction of monoclonal antibodies (MoAb). Origin lymphocytes of mice that produce antibodies againstC. cellulosae antigen, fused with myeloma cells (NS1). Results fusion of two cells produces hybrid cellscalled hybridomas; cells are then screened by ELISA test. Hybridoma cells that produce only MoAb, usedto produce large quantities in vitro. Characterization of MoAb against-C.cellulosae was tested by usingELISA and Western blotting. Mice were immunized with C.cellulosae antigen showed an immune responseproducing antibodies to C.cellulosae. Based on the results of fusion, produced a total of 51 hybridoma cellclones and after being screened, only three clones of hybridoma cells that produced MoAb against–C.cellulosae. MoAb produced, named after the hole where the growth of the ELISA micro plate, the BE6,BE7, and EE9. Characteristics of this MoAb capable of tracking cellulosae of fluid larvae and recognizeantigen protein bands with molecular weight 78kDa.
COMPARATIVE IMMUNE RESPONSES OF CHILDREN AFTER INTRADERMAL AND INTRAMUSCULAR RABIES VACCINATION Subawa, A. A. Ngurah; Sutirta Yasa, I Wayan Putu; Astawa, Nyoman Mantik
BALI MEDICAL JOURNAL Vol 3, No 3 (2014)
Publisher : BALI MEDICAL JOURNAL

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Abstract

Background: Rabies is a cause of death to people within 100% of Case Fatality Rate. Approximately 55.000 people died because of rabies each year, the vast majority of these deaths happen in Asia and Africa. This study aims to find out comparative immune responses of intradermal (ID) and intramuscular (IM) vaccination in children. Method: This was an experimental study to determine antibody response of ID and IM rabies vaccines with randomized pre and posttest control group design. ID and IM vaccination were carried out in 4 replication for each group. A number of 16 children were recruited for each group. Enzyme Linked Immunosorbent Assay (ELISA) was applied to determine titers antibody on day 0, 7, 21, and 28 after vaccination. Results: This study found that titer antibody induced by ID vaccination was lower than IM vaccination. However, the different is not statistically significant in both groups  (p > 0.05). Titers antibody on day 7 after vaccination were 3.08 ± 2.09 IU/ml intradermally and 4.22 ± 3.02 IU/ml intramuscularly. On day 21 and 28 after intradermal vaccination, titers antibody were 6.78 ± 3.52 IU/ml and 12.53 ± 5.92 IU/ml, respectively. Intramuscularly, antibody titers were 9.76 ± 4.86 IU/ml on day 21 and 14.98 ± 7.76 IU/ml on day 28. Conclusion: ID vaccination is safe and can be used as an alternative vaccination for rabies in human. In addition, 0, 7, 21 ID vaccination methods can be recommended for use to control rabies cases in Indonesia because that methods induce protective immune response.
EFFECT OF RESTING TIME ON PERIPHERAL BLOOD MONONUCLEAR CELL YIELDS Narayani, Inna; Niruri, Rasmaya; Astawa, Nyoman Mantik
International Journal of Biosciences and Biotechnology Vol 3 No 2 (2016)
Publisher : Faculty of Agriculture, Udayana University in cooperation with Asia-Oceania Bioscience and Biotechnology Consortium (AOBBC)

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Cryopreservation of PBMC (peripheral blood mononuclear cells) was done to preserve and analyze the number of PBMC derived from blood samples which come in at different time. The batch analysis wasperformed at the same time in order to reduce variations in results. The analysis on the cells numbers carried out after 1, 3, 6, 12, and 24 hours. Heparinized whole blood was collected from healthy subject by venipuncture, and stored at room temperature. Blood is processed by centrifugation in Ficoll density gradient following the established method of Balai Besar Veteriner Denpasar. Buffy coat layer was collected and washed twice with HBSS (Hank's balanced salt solution) and was counted in Turk’s solution. The cells were then dissolved in 1 ml of cold freezing medium containing 10% DMSO and 50% FBS (fetal bovine serum) and stored overnight at -80°C before storage in liquid nitrogen vessel for few weeks. The samples rapidly thawed in a water bath at 37°C and washed twice with PBS (phosphate buffered saline). The cells were stored in 4°C PBS and counted in Turk’s solution after 1, 3, 6, 12, and 24 hours. The results obtained were varied with a declining trend.
Kerusakan Hati Akibat Keracunan Alkohol Berulang pada Tikus Wistar (LIVER DAMAGE DUE TO ALCOHOL INTOXICATION REPEAT IN WISTAR RATS) Suaniti, Ni Made; Djelantik, Anak Agung Gede Sudewa; Suastika, Ketut; Astawa, Nyoman Mantik
Jurnal Veteriner Vol 13, No 2 (2012)
Publisher : Jurnal Veteriner

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The aims of this study was to determine the liver damage from alcohol intoxication in Wistar rats.The design used in this study was a randomized true experimental post test only control group design. Thestudy used 15 rats divided into 3 treatment groups each of which consists of 5 rats. The first group wasgiven distill water. The second group was given 5% alcohol, and the third group was given 20% alcohol. Ratswere treated with alcohol daily for six weeks. Biochemical markers were detected the levels of aldehydedehydrogenase (ALDH) in serum and histological changes in liver tissue. ALDH is a biochemical markerof a sensitive and specific ethanol after chronic alcohol administration. Blood sample was collected at 6and 24 hours after the last peroral administration of repeated alcohol treatment, and serum levels ofALDH was tested by enzyme linked immunosorbent assay (ELISA). The results showed that the levels ofALDH in the blood of alcohol treated Wistar rats significantly higher as compared to those of control rats.ALDH levels increased by 83.11% after administration of 5% alcohol and 112.05% after administration of20% taken after 6 hours of alcohol for 6 weeks. On samples taken after 24 hours, ALDH levels by 95.11%after administration of 5% alcohol and 86.79% after administration of 20% alcohol. Oral treatment with20% alcohol chronically was led to changes in the microscopic structure (necrosis) of liver tissue in Wistarrats. Liver tissue damage occured due to repeated use of alcohol is accompanied by increasing serum levelsof ALDH in Wistar rats.