I Gusti Ngurah Kade Mahardika

Kinetika Immunoglobulin Kuning Telur Antiparvovirus Anjing Pada Anjing (KINETICS OF ANTICANINE PARVOVIRUS YOLK IMMUNOGLOBULIN IN DOGS)

Kinetic study on Anti CPV IgY has been performed on six dogs aged 5-10 months. The IgY was injectedintravenously at dose of 21.4mg /10kg body weight. IgY levels in the blood were determined by ELISA. Aresearch was conducted to find out the kinetics of Anti CPV IgY in dogs blood. The kinetics of IgY wascalculated by using regression analysis to determine the association on the levels of IgY in serum againsttime at injection. The results showed that kinetic parameters were calculated based on first order kinetics.The constant elimination rate of IgY was at the range between 0.007 to 0.015 / h. IgY concentration in thedogs blood was from 0.746 to 0.992 mg / mL. The half-life of IgY was from 1.65 to 4.01 / d. Volumedistribution of IgY was between 21.47 to 28,55 / mL. Total IgY in the dog bodies (AUC) was from 42,60 to142,00 mg / mL.h. The duration of the IgY in the dog’s body was 3.08 to 8.51 days. Clearance time of IgY was0.15 to 0.50 mL / h. In conclusion the kinetics of anti CPV IgY in dog’s body follow one compartment andfirst order model, which are only distributed in the blood with the half-life at 2.5 days, and IgY has lesspossibility to accumulate in the body compared to the IgG.

Identification of Pathogenecity of Avian Influenza Virus Subtype H5N1 from Waterfowls Base on Amino Acid Sequence of Cleavage Site Hemagglutinin Protein

Identification of pathotype of Avian Influenza Virus (AIV) subtype H5N1 isolates is very important. This research aimed to identify the pathotype of AIV subtype H5N1 isolated from household waterfowls in West Java based on molecular markers of amino acid sequences of the Hemagglutinin (HA) cleavage site. Fragments of HA genes of 21 isolates were amplified using RT-PCR with a primer pair that flanking the cleavage site region, and sequenced with dideoxy-termination method with ABI automatic sequencer (Applied Biosystems). Multiple alignment of 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 of amino acid sequence, i.e QRERRRKKR (20 isolates) and QRESRRKKR (1 isolate). This finding indicates that all of the viruses isolated in this research were of highly pathogenic avian influenza (HPAI) strains. Keywords: cleavage site, waterfowls, HPAI

Molecular analysis of Hemagglutinin Gen of Highly Pathogenic Avian Influenza of H5N1 Subtype Isolated from Waterfowls

Avian influenza viruses (AIV) subtype H5N1 isolated from waterfowls in West Java pose the known characteristic of highly pathogenic strains, with polybasic amino acid sequence of cleavage site QRERRRKKR and QRESRRKKR. This research aimed to analyze the important domains of hemagglutinin (HA) gene of those isolates. Fragment of HA gene was amplified using RT-PCR method with specifically-designed primer pairs and sequenced using dideoxy termination method with ABI automatic sequencer (Applied Biosystems). Multiple alignment of nucleotide and deduced amino acid sequences were analyzed using ClustalW of MEGA-3.1 program. Some of biological domains of HA, i.e. antigenic sites, receptor binding pocket, and glycosylation sites of the isolates were polymorphic. The viruses also pose conserved glutamine (Q) and glisin (G) residues at the known receptor binding site, at the position 222 and 224 respectively. This findings clearly show that all AIV subtype H5N1 isolaed from waterfowl preservesthe α-2,3NeuAcGal avian receptor specificity. Key Words : Antigenic Sites, Glycosylation Sites, Receptor Binding Pocket, AIV H5N1, Waterfowls