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Identification of avian influenza virus of Indonesian isolates by reverse transcriptase polymerase chain reaction (RT-PCR) method Dharmayanti, N.L.P.I.; Damayanti, R; Wiyono, A; Indriani, R; ., Darminto
Indonesian Journal of Animal and Veterinary Sciences Vol 9, No 2 (2004)
Publisher : Indonesian Animal Sciences Society

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. Â Key words: Identification, avian influenza virus, RT-PCR, H5 subtype

Indonesian avian influenza viruses character in second wave epidemic Dharmayanti, N.L.P.I.; Indriani, R; Damayanti, R; Wiyono, A; Adjid, R.M.A.
Indonesian Journal of Animal and Veterinary Sciences Vol 10, No 3 (2005)
Publisher : Indonesian Animal Sciences Society

Second wave of epidemic avian influenza occurred from December 2004 until April 2005. In March 2005, the disease had infected some districts in South Sulawesi such as Wajo and Sopeng. More than 21 field isolates have been collected and identified as avian influenza virus subtype H5N1. In this study further characterized was undertaken for 14 isolates of avian influenza using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and sequencing in region of HA1 gene. It was then followed by genetic analysis to identify the mutation and phylogenetic relationship of the isolates. The study indicates that the Indonesia isolates collected in second wave epidemic are generally having a different group to the isolates group in 2003 and 2004. There is point mutation in the nucleotide sequence of the isolate collected at August 2004-March 2005, that is the replacement of adenine by guanine in the position of 195. Â Â Key Words: Avian Influenza Virus, Second Epidemic Wave, Mutation

Molecular characterization of Indonesia avian influenza virus Dharmayanti, N.L.P.I.; Damayanti, R; Indriani, R; Wiyono, A; Adjid, R.M.A.
Indonesian Journal of Animal and Veterinary Sciences Vol 10, No 2 (2005)
Publisher : Indonesian Animal Sciences Society

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). Â Â Key Words: Avian Influenza Virus, Molecular Characterization, Poultry, Indonesia

Protection of avian influenza (AI) vaccines for poultry against infection of field isolates A/Chicken/West Java/Smi-Pat/2006 and A/Chicken/West Java/Smi-Mae/2008 under laboratory condition Indriani, Risa; Dharmayanti, N.l.p.I.; Adjid, R.M.A.
Indonesian Journal of Animal and Veterinary Sciences Vol 16, No 2 (2011)
Publisher : Indonesian Animal Sciences Society

The aim of this research was to study level of protection of avian influenza (AI) commercial vaccines available in Indonesia (subtipe H5N1, H5N2 and H5N9) against infection of HPAI field isolates of A/Chicken/West Java/Smi-Pat/2006 and A/Chicken/West Java/Smi-Mae/2008. There were 7 commercial vaccines used in this study, the each vaccines were injected in to 3 weeks old of layer chichickenen intramuscularly. At 3 weeks after vaccination, ten chichickenens from each group were challenged separately with the A/Chicken/West Java/Smi-Pat/2006 and A/Chicken/West Java/Smi-Mae/2008 isolates intranasaly with dose 106 ELD50 per 0,1 ml per chicken. Ten unvaccinated chicken were included in the challenge test as control. The study demonstrate that the AI vaccines with subtipe H5N1 protected chicken (100%) against virus of A/Chicken/West Java/Smi-Pat/2006 and 90-100% against virus A/Chicken/West Java/Smi-Mae/2008. Viral shedding were not seen by 2 days post challenge. The AI vaccines with subtipe H5N2 protected chicken at 20-30% against virus of A/Chicken/West Java/Smi-Pat/2006 and protected chicken at 70-100% against virus of A/Chicken/West Java/Smi-Mae/2008. Viral shedding still detected at 8 days post challenge. The AI vaccines AI with subtipe H5N9 did not protect chicken (0%) against virus A/Chicken/West Java/Smi-Pat/2006 and protected chicken at 50% against virus A/Chicken/West Java/Smi-Mae/2008. Viral shedding still detected by 8 days post challenge. This study concluded that AI vaccines with subtipe H5N1 are better than other AI subtipe vaccines in preventing HPAI virus A/Chicken/West Java/Smi-Pat/2006 dan A/Chicken/West Java/Smi-Mae/2008 infections under laboratory condition. Key Words: Avian Influenza, Vaccine, Poultry, Chicken

Indonesian avian influenza viruses character in second wave epidemic N.L.P.I. Dharmayanti; R Indriani; R Damayanti; A Wiyono; R.M.A. Adjid
Jurnal Ilmu Ternak dan Veteriner Vol 10, No 3 (2005): SEPTEMBER 2005
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Second wave of epidemic avian influenza occurred from December 2004 until April 2005. In March 2005, the disease had infected some districts in South Sulawesi such as Wajo and Sopeng. More than 21 field isolates have been collected and identified as avian influenza virus subtype H5N1. In this study further characterized was undertaken for 14 isolates of avian influenza using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and sequencing in region of HA1 gene. It was then followed by genetic analysis to identify the mutation and phylogenetic relationship of the isolates. The study indicates that the Indonesia isolates collected in second wave epidemic are generally having a different group to the isolates group in 2003 and 2004. There is point mutation in the nucleotide sequence of the isolate collected at August 2004-March 2005, that is the replacement of adenine by guanine in the position of 195. Key Words: Avian Influenza Virus, Second Epidemic Wave, Mutation

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

Protection of avian influenza (AI) vaccines for poultry against infection of field isolates A/Chicken/West Java/Smi-Pat/2006 and A/Chicken/West Java/Smi-Mae/2008 under laboratory condition Risa Indriani; N.l.p.I. Dharmayanti; R.M.A. Adjid
Jurnal Ilmu Ternak dan Veteriner Vol 16, No 2 (2011): JUNE 2011
Publisher : Indonesian Center for Animal Research and Development (ICARD)

The aim of this research was to study level of protection of avian influenza (AI) commercial vaccines available in Indonesia (subtipe H5N1, H5N2 and H5N9) against infection of HPAI field isolates of A/Chicken/West Java/Smi-Pat/2006 and A/Chicken/West Java/Smi-Mae/2008. There were 7 commercial vaccines used in this study, the each vaccines were injected in to 3 weeks old of layer chichickenen intramuscularly. At 3 weeks after vaccination, ten chichickenens from each group were challenged separately with the A/Chicken/West Java/Smi-Pat/2006 and A/Chicken/West Java/Smi-Mae/2008 isolates intranasaly with dose 106 ELD50 per 0,1 ml per chicken. Ten unvaccinated chicken were included in the challenge test as control. The study demonstrate that the AI vaccines with subtipe H5N1 protected chicken (100%) against virus of A/Chicken/West Java/Smi-Pat/2006 and 90-100% against virus A/Chicken/West Java/Smi-Mae/2008. Viral shedding were not seen by 2 days post challenge. The AI vaccines with subtipe H5N2 protected chicken at 20-30% against virus of A/Chicken/West Java/Smi-Pat/2006 and protected chicken at 70-100% against virus of A/Chicken/West Java/Smi-Mae/2008. Viral shedding still detected at 8 days post challenge. The AI vaccines AI with subtipe H5N9 did not protect chicken (0%) against virus A/Chicken/West Java/Smi-Pat/2006 and protected chicken at 50% against virus A/Chicken/West Java/Smi-Mae/2008. Viral shedding still detected by 8 days post challenge. This study concluded that AI vaccines with subtipe H5N1 are better than other AI subtipe vaccines in preventing HPAI virus A/Chicken/West Java/Smi-Pat/2006 dan A/Chicken/West Java/Smi-Mae/2008 infections under laboratory condition. Key Words: Avian Influenza, Vaccine, Poultry, Chicken

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

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