N.L.P.I. Mayasari, N.L.P.I.
FKH IPB

Published : 5 Documents
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Journal : Indonesian Journal of Animal and Veterinary Sciences

Molecular analysis of hemaglutinin gene of Avian Influenza viruses isolated in 2012-2013 Kurniasih, Sussi Widi; Soejoedono, Retno D.; Mayasari, N.L.P.I.
Indonesian Journal of Animal and Veterinary Sciences Vol 20, No 2 (2015): JUNE 2015
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (399.055 KB) | DOI: 10.14334/jitv.v20i2.1166

Abstract

Avian Influenza virus (AIV) still plays as a major cause of the death in poultry in Indonesia and around the world. The aim of this research was to determine the pathogenicity and to analyze the phylogenetic and genetic distances of hemagglutinin gene of isolated AI viruses in Indonesia in 2012-2013 particularly from West Java, Central Java, and North Sumatra. Samples were obtained from poultry farms that suffered from AI outbreaks, were inoculated and propagated in ten days old specific pathogen free (SPF) embryonated chicken eggs. Harvested allantoic fluids at 5 days after inoculation were tested for hemagglutination activity. Positive allantoic fluids were further tested to determine the hemagglutinin and neuraminidase subtype using real-time reverse transcription polymerase chain reaction (RRT-PCR) and to be prepared for sequencing using reverse transcription polymerase chain reaction (RT-PCR). The sequence of hemagglutinin genes were analyzed for the amino acid pattern of the cleavage site region and the genetic distances and relationships of those viruses. The result indicated that all of the isolates are classified as HPAI with the pattern of cleavage site regions are QRESRRKKR and QRERRRKR. Six isolates are classified as H5N1 and 3 isolates are H5Nx. All of the isolates have close genetic relationship with the genetic distances less than 0.3 between one to another and also with several AI viruses that caused previous outbreaks in Indonesia. Key Words: Avian Influenza, Cleavage Site, Hemagglutinin, Pathogenicity, Phylogenetic
Newcastle Disease Virus infection study on duck and chicken in Subang district Panus, Aprizal; Setiyaningsih, S.; Mayasari, N.L.P.I.
Indonesian Journal of Animal and Veterinary Sciences Vol 20, No 2 (2015): JUNE 2015
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (706.973 KB) | DOI: 10.14334/jitv.v20i2.1168

Abstract

The objectives of this research were to study Newcastle Disease Virus (NDV) infection in Subang area and to examine the diversity of the circulating NDV. Swabs of cloacal and oropharynx, and serum were sampled from total of 393 chickens and 149 ducks in backyard farms and live bird markets located in 10 subdistricts. Screening of NDV in pool of 5-7 samples by real-time Reverse-Transcription Polymerase Chain Reaction (rRT-PCR) matrix (M) showed 19/67 (28.3%) cloacal and 8/67 (11.9%) pharyngeal pools of chicken samples; 18/67 (26.9%) of the pools excreted virus via cloaca and oropharynx, while the duck pools of 8/30 (26.7%) shed virus from cloaca. Virus isolation attempted on individual sample from positive pools yielded 18 isolates which the majority of the isolates showed homogeneous antigenic character, only some of these showed variations up to 2 Log2 with Lasota and 4 Log2 with Komarov antisera. Majority of isolates had a higher affinity to Komarov indicating their propencity to virulent strains. Pathogenicity examination using elution test showed 3 isolates virus were grouped to mesogenic strains and 15 isolates to velogenic strain, in agreement with rRT-PCR fusion results. HI test on 408 sera showed that NDV antibody was detected in 48 (12%) birds with titres ranging from 1 to 8 Log2; only about 13% of vaccinated chickens demonstrated protective antibody titre (≥3 Log2). Newcastle disease is still endemic in Subang with relatively low antigenic variation among circulating strains. Key Words: Newcastle Disease, rRT-PCR Detection, Detection of Virulence, Antigenic Diversity, Antibody
Newcastle Disease Virus infection study on duck and chicken in Subang district Panus, Aprizal; Setiyaningsih, S.; Mayasari, N.L.P.I.
Jurnal Ilmu Ternak dan Veteriner Vol 20, No 2 (2015): JUNE 2015
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (706.973 KB) | DOI: 10.14334/jitv.v20i2.1168

Abstract

The objectives of this research were to study Newcastle Disease Virus (NDV) infection in Subang area and to examine the diversity of the circulating NDV. Swabs of cloacal and oropharynx, and serum were sampled from total of 393 chickens and 149 ducks in backyard farms and live bird markets located in 10 subdistricts. Screening of NDV in pool of 5-7 samples by real-time Reverse-Transcription Polymerase Chain Reaction (rRT-PCR) matrix (M) showed 19/67 (28.3%) cloacal and 8/67 (11.9%) pharyngeal pools of chicken samples; 18/67 (26.9%) of the pools excreted virus via cloaca and oropharynx, while the duck pools of 8/30 (26.7%) shed virus from cloaca. Virus isolation attempted on individual sample from positive pools yielded 18 isolates which the majority of the isolates showed homogeneous antigenic character, only some of these showed variations up to 2 Log2 with Lasota and 4 Log2 with Komarov antisera. Majority of isolates had a higher affinity to Komarov indicating their propencity to virulent strains. Pathogenicity examination using elution test showed 3 isolates virus were grouped to mesogenic strains and 15 isolates to velogenic strain, in agreement with rRT-PCR fusion results. HI test on 408 sera showed that NDV antibody was detected in 48 (12%) birds with titres ranging from 1 to 8 Log2; only about 13% of vaccinated chickens demonstrated protective antibody titre (≥3 Log2). Newcastle disease is still endemic in Subang with relatively low antigenic variation among circulating strains.
Molecular analysis of hemaglutinin gene of Avian Influenza viruses isolated in 2012-2013 Kurniasih, Sussi Widi; Soejoedono, Retno D.; Mayasari, N.L.P.I.
Jurnal Ilmu Ternak dan Veteriner Vol 20, No 2 (2015): JUNE 2015
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (399.055 KB) | DOI: 10.14334/jitv.v20i2.1166

Abstract

Avian Influenza virus (AIV) still plays as a major cause of the death in poultry in Indonesia and around the world. The aim of this research was to determine the pathogenicity and to analyze the phylogenetic and genetic distances of hemagglutinin gene of isolated AI viruses in Indonesia in 2012-2013 particularly from West Java, Central Java, and North Sumatra. Samples were obtained from poultry farms that suffered from AI outbreaks, were inoculated and propagated in ten days old specific pathogen free (SPF) embryonated chicken eggs. Harvested allantoic fluids at 5 days after inoculation were tested for hemagglutination activity. Positive allantoic fluids were further tested to determine the hemagglutinin and neuraminidase subtype using real-time reverse transcription polymerase chain reaction (RRT-PCR) and to be prepared for sequencing using reverse transcription polymerase chain reaction (RT-PCR). The sequence of hemagglutinin genes were analyzed for the amino acid pattern of the cleavage site region and the genetic distances and relationships of those viruses. The result indicated that all of the isolates are classified as HPAI with the pattern of cleavage site regions are QRESRRKKR and QRERRRKR. Six isolates are classified as H5N1 and 3 isolates are H5Nx. All of the isolates have close genetic relationship with the genetic distances less than 0.3 between one to another and also with several AI viruses that caused previous outbreaks in Indonesia.