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N.L.P.I Dharmayanti
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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 Damayanti, R; Dharmayanti, N.L.P.I; Indriani, R; Wiyono, A; ., Darminto
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 (310.53 KB) | DOI: 10.14334/jitv.v9i3.409

Abstract

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
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

Abstract

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
Development inactivated vaccine prototype of avian influenza (AI) H5N1 local isolate and its application at laboratory level Indriani, R; Dharmayanti, N.L.P.I; Syafriati, T; Wiyono, A; Adjid, R.M.A
Indonesian Journal of Animal and Veterinary Sciences Vol 10, No 4 (2005)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (174.851 KB) | DOI: 10.14334/jitv.v10i4.458

Abstract

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
Monitoring of avian influenza cases based on the detection of viral antigen subtype H5N1 by immunohistochemical technique Damayanti, Rini; Dharmayanti, N.L.P.I; Indriani, R.; Wiyono, A.; Adjid, R.M.A
Indonesian Journal of Animal and Veterinary Sciences Vol 10, No 4 (2005)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (263.695 KB) | DOI: 10.14334/jitv.v10i4.1106

Abstract

Monitoring on the cases of Avian Influenza virus was conducted by detecting viral antigen subtype H5N1 usingimmunohistochemical technique. A total of 212 sampels of various avian tissues were collected from the Provinces of East Java(Districts of Madiun, Tulung Agung, Blitar and Kediri), West Java (Districts of Bogor, Bekasi, Cianjur and Sukabumi), Banten(Districts of Pandeglang and Tangerang) and DKI Jakarta. The sampels were collected four times i.e. June 2004, September2004, October 2004 and between January and February 2005. All sampels were stained using immunohistochemical technique.The antigen could be visualized clearly both in the intra-nuclear and intra-cytoplasmic areas of brain, comb, wattle, trachea,lung, heart, breast and thigh muscle, proventriculus, liver, spleen, kidney, intestine and ovary. A number of 39 of 212 cases(18.4%) have been catagorized as positives. The results show that monitoring of HPAI cases conducted in June and September2004 in the Provinces of West Java, Banten and East Java, none of the sampels were positive. However, monitoring of thedisease in September 2004 in the Province of Jakarta showed that AI virus antigen were detected in various organs of chickenfrom Jakarta. Furthermore, monitoring of the disease conducted between October 2004 and February 2005 revealed that AI virusantigen were also detected in chicken not only from Jakarta Provinces but also from Provinces of Banten and West Java. Basedon these results, it is concluded that between June and September 2004, HPAI infection were not found in areas where previousoutbreaks occured in the Provinces of Banten, West Java and East Java. However, the disease was spread in Jakarta Province inSeptember 2004 and subsequently to some districts in the Provinces of Banten and West Java. A part from this, anticipation ofdisease spread to currently AI-free areas should be considered as part of disease monitoring system.Key Words: Avian Influenza, H5N1, Monitoring, Immunohistochemistry, Poultry
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)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (310.53 KB) | DOI: 10.14334/jitv.v9i3.409

Abstract

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
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)

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

Abstract

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
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)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (174.851 KB) | DOI: 10.14334/jitv.v10i4.458

Abstract

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
Monitoring of avian influenza cases based on the detection of viral antigen subtype H5N1 by immunohistochemical technique Rini Damayanti; N.L.P.I Dharmayanti; R. Indriani; 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)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (263.695 KB) | DOI: 10.14334/jitv.v10i4.1106

Abstract

Monitoring on the cases of Avian Influenza virus was conducted by detecting viral antigen subtype H5N1 usingimmunohistochemical technique. A total of 212 sampels of various avian tissues were collected from the Provinces of East Java(Districts of Madiun, Tulung Agung, Blitar and Kediri), West Java (Districts of Bogor, Bekasi, Cianjur and Sukabumi), Banten(Districts of Pandeglang and Tangerang) and DKI Jakarta. The sampels were collected four times i.e. June 2004, September2004, October 2004 and between January and February 2005. All sampels were stained using immunohistochemical technique.The antigen could be visualized clearly both in the intra-nuclear and intra-cytoplasmic areas of brain, comb, wattle, trachea,lung, heart, breast and thigh muscle, proventriculus, liver, spleen, kidney, intestine and ovary. A number of 39 of 212 cases(18.4%) have been catagorized as positives. The results show that monitoring of HPAI cases conducted in June and September2004 in the Provinces of West Java, Banten and East Java, none of the sampels were positive. However, monitoring of thedisease in September 2004 in the Province of Jakarta showed that AI virus antigen were detected in various organs of chickenfrom Jakarta. Furthermore, monitoring of the disease conducted between October 2004 and February 2005 revealed that AI virusantigen were also detected in chicken not only from Jakarta Provinces but also from Provinces of Banten and West Java. Basedon these results, it is concluded that between June and September 2004, HPAI infection were not found in areas where previousoutbreaks occured in the Provinces of Banten, West Java and East Java. However, the disease was spread in Jakarta Province inSeptember 2004 and subsequently to some districts in the Provinces of Banten and West Java. A part from this, anticipation ofdisease spread to currently AI-free areas should be considered as part of disease monitoring system.Key Words: Avian Influenza, H5N1, Monitoring, Immunohistochemistry, Poultry
Co-Authors A Wiyono A. Wiyono Darminto . Darminto . L Parede R Damayanti R Indriani R. Indriani R.M.A Adjid Rini Damayanti Risa Indriani RISA INDRIANI T Syafriati