Abstract
Since 2015, Fowl adenovirus serotype 4 (FAdV-4) infection has caused serious economic losses to the poultry industry worldwide. We isolated and identified the FAdV-4 strain NP, from infected chickens on a layer farm, using chicken embryo allantoic cavity inoculation, electron microscopy, viral genome sequencing, and regression analysis. To explore the pathogenesis of FAdV-4 infection, we conducted transcriptome sequencing analysis of the liver in chickens infected with FAdV-4, using the Illumina® HiSeq 2000 system. Two days after infection with the FAdV-4 NP strain, 13,576 differentially expressed genes (DEGs) were screened in the liver, among which, 7,480 were up-regulated and 6,096 were down-regulated. Gene ontology (GO) analysis indicated that these genes were involved in 52 biological functions. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that those DEGs were involved in 33 pathways. We then focused on the KEGG pathway of phagosome and found that mRNA levels of the 25 DEGs in that pathway were up-regulated, and seven DEGs were down-regulated. Real-time quantitative polymerase chain reaction (qPCR) confirmed the accuracy and reliability of these findings. Moreover, 24 h after LMH cells were infected with FAdV-4, the mRNA levels of F-actin, Rab7, TUBA, and DVnein were significantly increased. These four genes were all subsequently silenced by RNA interference, and viral replication of FAdV-4 was then significantly down-regulated. These findings demonstrate the isolation and identification of the FAdV-4 NP strain, and the DEGs in KEGG pathway of phagosome were utilized by FAdV-4 to benefit its infection.
Highlights
Based on the genomic profiles, Fowl adenovirus (FAdV) is divided into three groups (I–III) and 12 serotypes (Hess, 2000)
We focused on the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway of phagosome and used quantitative polymerase chain reaction (qPCR) and ribonucleic acid (RNA) interference to determine whether the KEGG pathway of phagosome was used to facilitate FAdV serotype 4 (FAdV-4) invasion
The hematoxylin and eosin (H&E) stains were purchased from Nanjing Jiancheng Technology Co., Ltd. (Nanjing, China), whereas the Dulbecco’s modified Eagle medium (DMEM), fetal bovine serum, and the real-time fluorescent quantitative polymerase chain reaction kit were purchased from TaiJing Biotechnology Co., Ltd. (Xiamen, China)
Summary
Based on the genomic profiles, Fowl adenovirus (FAdV) is divided into three groups (I–III) and 12 serotypes (Hess, 2000). It is a non-encapsulated, double-stranded DNA virus (Griffin and Nagy, 2011). Infection with different serotypes of group I FAdV leads to different clinical symptoms. In 1987, FAdV-4 was first reported in Pakistan, and in India (Mittal et al, 2014) and South Korea (Kim et al, 2008), among other countries. Before 2014, Intracellular Trafficking of Fowl Adenovirus Serotype 4 only sporadic cases had been reported in China. Since 2015, FAdV-4 infection has caused a major outbreak in China. Little is known about the pathogenic mechanisms of FAdV-4 infection
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