We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
内蒙古牛呼吸道合胞体病毒的分离鉴定及其全基因组分析.
- Authors
黄金田; 李 静; 丁玉林; 王文龙; 高 娃; 王金玲
- Abstract
In order to determine the pathogens responsible for the outbreak of respiratory disease in Holstein calves aged 3-6 months in Inner Mongolia and analyze its genome sequence. In this study, dead calve was dissected and observed for dissected lesions of each organ, and histopathological sections of each tissue were prepared and observed for histopathological lesions. The autopsy showed that the deep red shiny, muscular-like solid lesions in the lungs, hypertrophy of the heart, fine nodular greyish-white foci scattered on the surface of the liver, and fine deep red haemorrhagic dots scattered on the surface of the kidneys. Histopathological showed typical interstitial pneumonia with massive proliferation and infiltration of macrophages and lymphocytes in the lumen of most fine bronchioles at all levels and in the alveolar lumen, occasionally syncytiotrophoblast, reddish-stained virus inclusion bodies of varying sizes within the plasma of the epithelial cells of some of the mucous membranes of all bronchioles at all levels; metamorphic hepatitis with slight fibrosis and small necrotic foci, acute inflammatory splenomegaly, chronic sclerosing glomerulonephritis with multiple small haemorrhagic foci. RT-PCR results showed a target band of about 600bp, which was subjected to sequencing. Alignment of the target gene with the sequences available from GenBank showed that the highest homology (98.42%) is the BRSV US isolate USII/SI (KU159366), indicating the calf was infected with BRSV. The lung tissue grinding supernatants were inoculated into Madin-Darby Bovine Kidney (MDBK) cells, and a typical cytopathic effect was observed after three blind passages. BRSV was confirmed positive by RT-PCR detection, further confirming the successful isolation of BRSV, which was designated as IM BRSV-39. The complete genome sequence of IM BRSV-39 was amplified in segments, sequenced and spliced, revealing the full-length genome of the BRSV is 15130bp. The whole genome sequence, G and F gene sequence of IM BRSV-39 were analyzed for homology and mutation sites using MegAlign; the genetic evolutionary relationships of isolate were analyzed by constructing a genome-wide evolutionary tree using MEGA 11 software. The results showed that the whole genome sequence, G and F gene sequences of IM BRSV-39 had the highest homology and were in the same evolutionary branch with BRSV subtype III isolates, confirming that IM BRSV-39 belonged to BRSV subtype III. Compared with the reference BRSV III, the IM BRSV-39 had 7 and 12 mutations in the G and F gene, respectively. In addition, four amino acid substitutions occurred in G protein, of which L13H was in the cytoplasmic structural domain of the G protein (aal-aa37); Y127H, I139T, P177L, A186T are in the extracellular domain of G protein (aa66-aa257). Five amino acid substitutions were observed in F protein, in which M4T, M7T, and S14C are in the signal peptide region of the N-terminus of F protein (aal-aa26). In this study, we detected and isolated one strain of BSRV and revealed the genetic evolutionary characteristics of the viral genome, laying the foundatio.
- Subjects
WHOLE genome sequencing; PEPTIDES; G proteins; CELLULAR inclusions; VIRAL genomes
- Publication
Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao, 2024, Vol 46, Issue 5, p476
- ISSN
1008-0589
- Publication type
Article
- DOI
10.3969/j.issn.1008-0589.202312014