1.Hexon and fiber gene changes in an attenuated fowl adenovirus isolate from Malaysia in embryonated chicken eggs and its infectivity in chickens
Norfitriah M SOHAIMI ; Mohd H BEJO ; Abdul R OMAR ; Aini IDERIS ; Nurulfiza M ISA
Journal of Veterinary Science 2018;19(6):759-770
Fowl adenovirus (FAdV) is distributed worldwide and causes economic losses in the poultry industry. The objectives of this study were to determine the hexon and fiber gene changes in an attenuated FAdV isolate from Malaysia in specific pathogen-free chicken embryonated eggs (SPF CEE) and its infectivity in commercial broiler chickens. SPF CEE were inoculated with 0.1 mL FAdV inoculum via the chorioallantoic membrane (CAM) for 20 consecutive passages. The isolate at passage 20 (E20), with a virus titer of 10(8.7)TCID₅₀/mL (TCID₅₀, 50% tissue culture infective dose), was inoculated (0.5 mL) into one-day-old commercial broiler chicks either via oral or intraperitoneal routes. The study demonstrated that 100% embryonic mortality was recorded from E2 to E20 with a delayed pattern at E17 onwards. The lesions were confined to the liver and CAM. Substitutions of amino acids in the L1 loop of hexon at positions 49 and 66, and in the knob of fiber at positions 318 and 322 were recorded in the E20 isolate. The isolate belongs to serotype 8b and is non-pathogenic to broiler chickens, but it is able to induce a FAdV antibody titer. It appears that molecular changes in the L1 loop of hexon and the knob of fiber are markers for FAdV infectivity.
Adenoviridae
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Amino Acids
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Chickens
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Chorioallantoic Membrane
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Eggs
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Fowl adenovirus A
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Liver
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Malaysia
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Mortality
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Ovum
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Poultry
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Serogroup
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Specific Pathogen-Free Organisms
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Viral Load
2.Identification of a non-essential region for replication of fowl adenovirus QU strain.
Jin SUN ; Qiuyan LI ; Yunlong LI ; Bing HUANG ; Minxun SONG ; Xinhua LI
Chinese Journal of Biotechnology 2008;24(7):1263-1267
The avirulent QU strain of fowl adenovirus, a member of duck adenovirus type 1, could be a potential vector in recombinant vaccine development. To identify a non-essential region for replication of QU virus, a 3.4 kb fragment near the E4 region of QU virus genome was amplified by PCR to construct a plasmid pADGFP, in which ORF1, ORF8 and ORF9 was replaced with a system expressing enhanced green fluorescence protein. Further, a recombinant virus rQUGFP was constructed by homologous recombination after pADGFP and QU virus were co-transfected into chick embryo fibroblast. The one step growth curve of the rQUGFP was found to be identical with that of parent QU virus and the TCID50 titers of different generation recombinants maintained stable. These findings suggest that the region including ORF1, ORF8 and ORF9 of QU virus genome is dispensable for virus replication, and the foreign gene inserted into virus genome can be efficiently and stably expressed. The work lays the foundation for further studies of developing this virus as a vector of recombinant vaccine.
Adenovirus E4 Proteins
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genetics
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immunology
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Animals
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Fowl adenovirus A
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classification
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genetics
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Genes, Viral
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genetics
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Genetic Vectors
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genetics
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Open Reading Frames
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genetics
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Recombination, Genetic
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Transfection
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Vaccines, Synthetic
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biosynthesis
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genetics
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immunology
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Viral Vaccines
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biosynthesis
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genetics
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immunology
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Virus Replication