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Identification of Pathogenecity of Avian Influenza Virus Subtype H5N1 from Waterfowls Base on Amino Acid Sequence of Cleavage Site Hemagglutinin Protein R., Susanti; Soejoedono, Retno D.; K Mahardika, Gusti Ngurah I; Wibawan, Wayan T I; Suhartono, Maggy T
Indonesian Journal of Biotechnology Vol 13, No 2 (2008)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (635.287 KB)

Abstract

Identification of pathotype of Avian Influenza Virus (AIV) subtype H5N1 isolates is very important. Thisresearch aimed to identify the pathotype of AIV subtype H5N1 isolated from household waterfowls in West Javabased on molecular markers of amino acid sequences of the Hemagglutinin (HA) cleavage site. Fragments of HAgenes of 21 isolates were amplified using RT-PCR with a primer pair that flanking the cleavage site region, andsequenced with dideoxy-termination method with ABI automatic sequencer (Applied Biosystems). Multiple alignmentof nucleotide and their deduced amino acid sequence were analyzed using ClustalW from MEGA 3.1 program.The result shows that all H5N1 isolates (21 isolates) possess polybasic cleavage sites with 2 patterns ofamino acid sequence, i.e QRERRRKKR (20 isolates) and QRESRRKKR (1 isolate). This finding indicates that all ofthe viruses isolated in this research were of highly pathogenic avian influenza (HPAI) strains.Keywords: cleavage site, waterfowls, HPAI
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
L3 Populations in Laying Hens Infected with 6,000 L2 of Ascaridia galli D, Darmawi; Balqis, Ummu; Tiuria, Risa; Soejoedono, Retno D.; Pasaribu, Fachriyan H.
Jurnal Kedokteran Hewan Vol 1, No 2 (2007): September
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v1i2.3122

Abstract

The aim of the present study was to determine the survival of L3 populations in intestine ofchickens exposed to experimental Ascaridia galli infection. Nature female adult worm were obtained fromlumen of village chickens in a comercial abattoir in Bogor. The eggs (L1) obtained from uteri female adultworms were incubated in sterile aquadestilata at room temperature for 10-20 days developed embrionatedeggs (L2). Five groups (A-D) of 80 head chickens were infected with, 6000 L2 A. galli respectively. Thechickens of group A were infected six times with dose of each 1,000 L2 with an interval of one hour. Thechickens of group B were infected three times with dose of each 2,000 L2 with an interval of two hours.The chickens of group C were infected six times with dose of each 3,000 L2 with an interval of three hours. The chickens of group D were infected one time with single dose 6,000 L2. A. galli L3 were recovered from intestines of 80 heads chickens seven days after oesophagus inoculation with 6,000 L2.The result showed that total 702,000 L1 and 628,000 L2 collected from 124 A. galli female adult worms.The percentage of L1 developed L2 is 89.46% and L2 developed L3 is 11.27%. Significant survival of L3higher populations in intestine of chickens observed only in the group D. The results indicated thatchickens infected high dose of A. galli caused the decrease of host defence against ascaridiosis. Keywords: Ascaridia galli, embrionated eggs, larvae