SEROPREVALENSI PENYAKIT AVIAN INFLUENZA SUBTIPE H5N1 PADA AYAM BURAS DI PASAR BERINGKIT DAN GALIRAN, BALI

Avian Influenza (AI) is a strategic communicable and zoonotic disease. The cause is a virus with Highly Pathogenic Avian Influenza (HPAI) subtype H5N1. The poultry market has important roles in the preservation, propagation, and spreads of the Avian Influenza (AI) virus from poultry to other species and humans. The purpose of this study was to determine the level of seroprevalence of Avian Influenza (H5N1) in free-range chickens at the Beringkit and Galiran market. A total of 120 free-range chickens were used as a sample. They have taken 60 serum samples per market. Serum removal is made from 5 merchants under the provisions of 3 samples from merchants who sell 6 to 10 free-range chickens. The sampling period is carried out for 2 months every 2 weeks 4 times. Sample testing was performed at the Denpasar Veterinary Centre with Haemagglutination (HA) and Haemagglutination Inhibition (HI) as barriers. The data titer of the antibodies obtained was analyzed by Non-Parametric Statistic Test Chi-Square (χ2) using IBM SPSS for windows. The results of the study showed that the AI subtype of the H5N1 subtype in both Beringkit Markets is 3.3% and Galiran Market is 6.7%, with seroprevalence in the two markets of 5.0% which is statistically not dissimilar (P<0.05). To prevent the transmissions of AI disease at the Beringkit Market and Galiran is recommended for vaccination and more attention to the market management and the free-range chicken maintenance system

PENEGUHAN DIAGNOSIS PENYAKIT NEWCASTLE DISEASE LAPANG PADA AYAM BURAS DI BALI MENGGUNAKAN TEKNIK RT-PCR

Penelitian ini bertujuan mendiagnosis kasus newcastle disease (ND) lapangan pada ayam bukan ras yang bersifat akut melalui hasil pemeriksaan di Laboratorium Patologi dan Laboratorium Biomedik Fakultas Kedokteran Hewan Universitas Udayana tahun 2008-2009. Sebanyak sepuluh ekor sampel ayam buras telah diperiksa. Gejala klinis yang teramati meliputi: anoreksia, lesu, bersin, batuk, dan diare putih kehijauan dengan diagnosis sementara sebagai penyakit ND yang bersifat akut. Sampel diambil dari organ yang mengalami perubahan patognomonis seperti pada proventrikulus, ventrikulus, seka tonsil, paru-paru dan otak. Perbanyakan virus menggunakan telur ayam bertunas (TAB) umur 10 hari melalui ruang alantois dan diinkubasikan pada inkubator telur suhu 37° C selama 3 hari. Koleksi cairan alantois dilakukan pada hari ke-3, selanjutnya diidentifikasi dengan uji serologi hemaglutinasi (Haemaglutination-Inhibition Test/HA/HI) dengan teknik mikrotiter baku dan dikonfirmasi dengan uji reverse transcriptase-polymerase chain reaction (RT-PCR) menggunakan primer FNDIFP (5’-CCCCGTTGGAGGCATAC-3’) dan FNDIBP (5’-TGTTGGCAGCATTTTGATTG-3’). Hasil penelitian menunjukkan bahwa semua sampel kasus ayam bukan ras yang diperiksa positif terinfeksi oleh virus penyakit ND akut. Kajian ini mengindikasikan bahwa penyakit ND di Bali masih bersifat endemis

Seroprevalence and detection of H5N1 avian influenza virus in local chickens in Tabanan Regency, Bali, Indonesia

Aim: Avian Influenza (AI) is a zoonotic disease that causes death in poultry and humans. Monitoring the virus needs to be carried out continuously to prevent outbreaks of the disease. Seroprevalence and detection of H5N1 and H9N2 AI virus antigen were intended to monitor the presence of viruses in local chickens in Tabanan, a Regency of the Indonesian island Province of Bali. The research aims were to detect the presence of H5N1 AI virus, and to know the distribution of this virus in Tabanan Regency. Materials and Methods: Research located in six districts of Tabanan regency namely Baturiti, Penebel, Marga, Kediri, Tabanan, and Kerambitan. A total of 1,398 local chickens that never been vaccinated with AI were randomly sampled in this study. The samples collected were serum, cloacal and tracheal swabs. Serum samples were tested with hemagglutination inhibition (HI) assay. While samples of cloacal and tracheal swabs were isolated in 9-day-old germinated chicken eggs, followed by hemagglutination assay and RT-PCR test using H5N1 and H9N2 primers. Results: AI seroprevalence in local chickens in Tabanan Regency was 1% with the distribution in each district as follows; Penebel 1.6%, Kerambitan 1.2%, Marga 1%, while Tabanan, Kediri, and Baturiti 0.7% each. H5N1 AI virus was detected in 11 samples,&nbsp; i.e. five in Marga district and three in Penebel district, two in Kediri, and one in Tabanan, while the H9N2 AI virus was not detected. Conclusion: These results indicate that H5N1 AI virus may still circulate in local chickens in Tabanan Regency, Bali

The Production and Use of Monoclonal Antibodies for the Detection of Avian Influenza Antigen in the in Infected Chickens

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

Seroprevalensi Penyakit Egg Drop Syndrome pada Itik di Desa Tumbak Bayuh, Kecamatan Mengwi, Badung, Bali

Egg Drop Syndrome can cause detrimental impacts on breeders due to reducing production and quality of theaffected eggs.This study aim was to determine the seroprevalece ofantibody against Egg Drop Syndrome (EDS) virus in ducks. Antibody titers examination was done from 75blood samplesof ducks thathave not been vaccinatedagainstEDS virus. The duck samples were collected from Tumbak Bayuh Village, Mengwi, Badung. Serological examination was held at the Virology Laboratory,Faculty of Veterinary Medicine, Udayana University by usingHaemagglutination Inhibition (HI) test. HI test results showed that 24 samples were positive contained antibody of the EDS,while 51 samples were negative. Range of EDS antibody titer observed was from24 to 27 HI units.  This results indicates that the ducks have protective antibody titer against EDS virus. The positive serum samples were also tested using HI test against Newcastle Disease (ND) virus with negative result. It can be concluded that the ducks in Tumbak Bayuh Village, Mengwi, Badung have 32% of antibody titer against EDS virus whichmight result from being exposed by EDS virus naturally

ASSESSMENT OF VIRAL CONTENT IN NEWCASTLE DISEASE VACCINE OBTAINED FROM TWO DIFFERENT POULTRY SHOPS USING PRIMARY CHICKEN FIBROBLAST CELL CULTURE

Newcastle Disease (ND) is still endemic in Indonesia, characterized by its your-round occurrence. Many measures have been adopted by the government to prevent the spread of the disease including vaccination using both active and inactive vaccine. The quality of vaccine is influenced by its viral content which determines the success of ND vaccination in chicken flock. The viral content in ND vaccine can be determined by measuring its eggs lethal dose-50 (ELD50) or Tissue Culture Infective Dose-50 (TCID50) and the minimum viral content considered to be appropriate for active ND vaccine is 66,5/single dose. This study was conducted to find out the viral content of ND vaccine marketed in some poultry shops. ND vaccine of  LaSota strain were obtained from 2 different poultry shops and the viral content was determined in Chicken embryo fibroblast (CEF). The vaccines were reconstituted vaccine diluent and diluted serially in 10-fold diluted. Each dilution was inoculated into 4 wells of confluent CEF cultured in 96 well microplate. The TCID50 was then calculated by Reed and Muench method. The TCID50 of each vaccine was determined 4 times (4 replications). The result showed that the titer of the virus in the vaccine were 66,7and 67 TCID50/per dose which mean that both vaccines were still above the minimum standard of viral content recommended by some workers

Peran Coding dan Non-Coding Region dari Gen Polimerase Kompleks Dalam Adaptasi Virus Avian Influenza

Avian influenza (AI) is an avian diseases that can cause deadly humaninfection. The disease is caused by Orthomyxoviridae virus, genus influenzavirus type A,subtype H5N1. Pathogenicity of AI virus is polygenic, which means AI virus isdetermined by all the genes. Here the role of coding region (CR) and non-coding region(NCR) of polymerase gene complex is critically reviewed. The coding region of thepolymerase genes can be explained as follows. The amino acid position 627 PB2 gene isa factor adaptation of AI viruses in mammals. Polymerase basic-1 (PB1) protein acts as acentral activity of the enzyme catalyzing the viral polymerase. Polymerase Basic-1 bindsto the terminal end of the vRNA and the cRNA and shows endo-nuclease activity. Theactivity has been identified on the E508, E519, and D522. Polymerase Acid (PA) proteinsplay a role in supporting the biological activity of the polymerase gene complex, but itsmechanism of action in transcription, replication has not been disclosed as clear. Anotherrole of the PA protein is forming a complex of RNA polymerase and express anproteolytic role of cell proteins that suppress cell division. Polymerase acid gene has alsoserine protease activity has been identified at position S624. The non-coding regionmight also play role in the pathogenecity of influenza virus. The results of sequenceanalysis of non-coding region (NCR) at 5’-end of polymerase gene complex of AI virussubtype H5N1 from poultry and pigs in Indonesia showed that the NCR genes PB2, PB1and PA are homogeneous, whereas there are variants of the PB1 gene isolates from ducksA / Duck / Badung, 2006. Occurrence of deletions, insertions, and mutations in the NCRand CR polymerase complex genes may likely lead to genetic changes in viruses whichpotentially also change the nature of biology of AI virus. The study on the 3’-end of thegenes needs to be carried out

Seroprevalensi Penyakit Flu Burung (Avian Influenza) pada Ayam Kampung di Kerta, Payangan, Gianyar, Bali

Desa Kerta berbatasan dengan Kecamatan Kintamani yang merupakan sentra industri peternakan ayam petelur. Penyakit AI pernah dilaporkan ditemukan pada peternakan ayam petelur di Kintamani. Lalu lintas perdagangan ayam pedaging maupun ayam petelur dari Kintamani ke Denpasar melewati desa Kerta. Populasi ayam kampung di desa Kerta cukup tinggi yakni 486.863 pada tahun 2015. Ayam kampung di Desa Kerta dipelihara secara ekstensif. Hal tersebut berpotensi besar dalam penularan virus flu burung (Avian Influenza = AI). Tujuan penelitian ini untuk mengetahui seroprevalensi penyakit AI pada ayam kampung di Desa Kerta, Kecamatan Payangan, Kabupaten Gianyar, Bali. Sampel penelitian sebanyak 80 ekor ayam kampung yang belum pernah divaksin dan dipelihara secara ekstensif. Lokasi penelitian di empat dusun desa Kerta yakni Dusun Pilan, Dusun Kerta, Dusun Buhu, dan Dusun Marga Tengah. Sampel serum diambil secara acak dari masing-masing dusun sebanyak 20 ekor. Pengujian dilakukan dengan uji haemaglutinasi (HA/HI). Analisis data dilakukan secara deskriptif. Hasil analisis data serologi diperoleh prevalensi flu burung pada ayam kampung di Dusun Pilan sebesar 5%, Dusun Kerta 0%, Dusun Buhu 5%, dan Dusun Marga Tengah 0%. Tingkat seroprevalensi flu burung di Desa Kerta sebesar 2,5% dengan titer antibodi 22 HI unit dan 24 HI unit. Fakta tersebut menunjukkan bahwa ayam yang disampling pernah terpapar virus flu burung secara alami. Disarankan agar ayam kampung di Desa Kerta dan sekitarnya divaksinasi untuk meningkatkan titer antibodi

Titer Antibodi Ayam Petelur Pascavaksinasi Avian Influenza Pada Peternakan Komersial Di Desa Denbantas, Kecamatan Tabanan

Penelitian ini bertujuan untuk mengetahui respon imun sekunder pascavaksinasi kedua AI pada peternakan komersial di Desa Denbantas, Kecamatan Tabanan. Sampel penelitian dipilih secara acak terhadap 10 ekor ayam petelur dari total 200 ekor. Sampel diambil sebanyak tiga kali yakni sekali pravaksinasi dan dua kali pascavaksinasi pada peternakan komersial di Desa Denbantas, Kecamatan Tabanan. Pemeriksaan titer antibodi AI dilakukan dengan uji serologi Haemaglutination Inhibition (HI). Nilai titer antibodi selanjutnya dianalisis menggunakan uji sidik ragam univariate dilanjutkan dengan uji Beda Nyata Terkecil (BNT), uji Duncan, dan analisis regresi. Hasil penelitian menunjukkan terjadi peningkatan titer antibodi yang signifikan setiap minggu pascavaksinasi. Rerata titer antibodi pravaksinasi sebesar 0 HI log 2. Rerata titer antibodi ayam petelur pascavaksinasi kedua pada minggu ke-2 yaitu 4,6 HI log 2 dan 8,3 HI log 2 pada minggu ke-3. Periode pengambilan serum berpengaruh sangat nyata (P<0,01) terhadap peningkatan titer antibodi AI pascavaksinasi setiap minggunya. Disarankan untuk memperhatikan waktu vaksinasi kedua dilakukan sebelum titer antibodi primer mencapai 0 HI log 2