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Detection of avian influenza virus H5N1 subtype in organs of chicken affected by higly pathogenic avian infuenza in East and West Java by using immunohistochemical technique R Damayanti; N.L.P.I Dharmayanti; R Indriani; A Wiyono; Darminto .
Jurnal Ilmu Ternak dan Veteriner Vol 9, No 3 (2004): SEPTEMBER 2004
Publisher : Indonesian Center for Animal Research and Development (ICARD)

The study was conducted to detect antigen H5N1 of highly pathogenic Avian Influenza (HPAI) virus in various farms in East and West Java. The immunohistochemical technique was applied due to Hematoxilin-eosin (H&E) staining was impossible to visualize the antigen in tissue. Immunohistochemical staining was applied for some visceral organs collected from the areas where the outbreaks occurred in September-October 2003. The specimens were processed as histopathological paraffin blocks using standard method. The blocks that were suspected to have antigen H5N1 were cut and rabbit antisera to H5N1 produced from the local isolate was applied as the primary antibody. Biotinylated secondary antibody and avidin biotin peroxidase from a commercial kit were administered. The antigen present in the tissues were visualized by adding a substrate called Amino Ethyl Carbazole (AEC) resulting in reddish brown colour. This immunostaining proved to be accurate and reliably quick method to detect H5N1 antigen present in the avian tissues. In conclusion, the outbreak of bird flu was caused by H5N1 strain and the antigen could be found in wattles, combs, brain, trachea, lungs, heart, proventriculus, liver, spleen, kidney and ovary. Key words: Highly pathogenic Avian Influenza (HPAI), chicken, H5N1, outbreak, immunohistochemistry

Molecular characterization of Indonesia avian influenza virus N.L.P.I. Dharmayanti; R Damayanti; R Indriani; A Wiyono; R.M.A. Adjid
Jurnal Ilmu Ternak dan Veteriner Vol 10, No 2 (2005): JUNE 2005
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Avian influenza outbreaks in poultry have been reported in Java island since August 2003. A total of 14 isolates of avian influenza virus has been isolated from October 2003 to October 2004. The viruses have been identified as HPAI H5N1 subtype. All of them were characterized further at genetic level and also for their pathogenicity. Phylogenetic analysis showed all of the avian influenza virus isolates were closely related to avian influenza virus from China (A/Duck/China/E319-2/03(H5N1). Molecular basis of pathogenicity in HA cleavage site indicated that the isolates of avian influenza virus have multiple basic amino acid (B-X-B-R) indicating that all of the isolates representing virulent avian influenza virus (highly pathogenic avian influenza virus). Key Words: Avian Influenza Virus, Molecular Characterization, Poultry, Indonesia

Detection of antibody responses by using haemagglutination inhibiton test and the protection titer of avian influenza virus H5N1 subtype Risa Indriani; N.L.P.I Dharmayanti; A Wiyono; Darminto .; L Parede
Jurnal Ilmu Ternak dan Veteriner Vol 9, No 3 (2004): SEPTEMBER 2004
Publisher : Indonesian Center for Animal Research and Development (ICARD)

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 N.L.P.I. Dharmayanti; R Damayanti; A Wiyono; R Indriani; Darminto .
Jurnal Ilmu Ternak dan Veteriner Vol 9, No 2 (2004): JUNE 2004
Publisher : Indonesian Center for Animal Research and Development (ICARD)

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

Development inactivated vaccine prototype of avian influenza (AI) H5N1 local isolate and its application at laboratory level R Indriani; N.L.P.I Dharmayanti; T Syafriati; A Wiyono; R.M.A Adjid
Jurnal Ilmu Ternak dan Veteriner Vol 10, No 4 (2005): DECEMBER 2005
Publisher : Indonesian Center for Animal Research and Development (ICARD)

A preliminary study related on vaccine safety and vaccination effectivity for controlling avian influenza (AI) subtype H5N1 was carried out at Virology Laboratorium, Indonesian Veteriner Institute, Bogor. A Prototype of inactivated vaccine was made using AI H5N1 local isolate (A/Chicken/West Java/67-2/2003). The vaccine was then tested for safety and protection in DOC of layers. Antibody response, protection and shedding virus challenge were observed in the experiment. Result showed that the vaccine was saved and protected against virulent viral challenge. Efective vaccination was achieved at 3 weeks chicken old started with low level of antibody. Antibody titre increased gradually and reached the top at 8 weeks post vaccination. Challenge test using AI virulent at the age of 4 and 8 weeks post vaccination showed that the vaccine gave high protection (90%). Viral shedding was not longer expressed than 7 days after challenge. It is concluded that this prototype is a satisfied AI vaccine in laboratory level. Key Words: Vaccine, Avian Influenza, H5N1, HPAI

Histopathological features of Marek’s disease infections in broiler chicken in Districts of Tasikmalaya and Ciamis West Java R Damayanti; A Wiyono
Jurnal Ilmu Ternak dan Veteriner Vol 8, No 4 (2003): DECEMBER 2003
Publisher : Indonesian Center for Animal Research and Development (ICARD)

An outbreak of Marek’s disease was reported to occur in broiler chicken in Districts of Tasikmalaya and Ciamis. A total number of 58 tissues samples of broiler chicken were collected from 7 flocks of commercial broiler chicken farms in both Districts. The disease affected broiler chicken aged 17 to 24 days. Those chickens had been vaccinated to Newcastle Disease (ND) and at age of 10 days had been vaccinated to Gumboro using blended bursa of fabricius. Tissue samples were fixed in 10% of buffered neutral formalin (BNF) prior to haematoxilin and eosin (H and E) stain using standard procedures. Histopathological features show that out of 58 samples, 32 (55.2%) were infected by Marek’s Disease (19.0% were infected by Marek’s Disease, 20.1% were infected by Marek’s Disease and Gumboro, 16.1% Marek’s Disease and other infections), whereas 44.8% were infected by Gumboro alone or accompanied by other infections, ND and Colibasillosis. The study reveals that Marek’s Disease infection in broiler chicken tends to be mild i.e. infiltration of neoplastic cells (lymphoid, pleomorphic) in proventriculus, intestine, spleen, livers and bursa of fabricius. In addition to this, there were mild non-supurative inflammation in heart, lung, peripheral nerve and brain, as well as a severe demyelination in brain. It is concluded that the histopthological features confirm the diagnosis of Marek’s Disease. Key words: Histopathology, Marek’s disease, broiler chicken, Districts of Tasikmalaya and Ciamis (West Java)

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