| Bovine enterovirus(BEV)infection is an infectious disease caused by bovine enterovirus within the enterovirus genus of picornaviridae,which is characterized by gastrointestinal and respiratory symptoms and causes serious economic losses to cattle industry.BEV infection has been reported in many countries and regions since it was first reported by Moll et al.in 1959.Li et al isolated the first strain of F enterovirus in China in 2011.Xing et al isolated the first enterovirus E strain of HY12 in China from cattle with severe diarrhea in Jilin Province in 2012.In recent years,with the rapid development of domestic cattle industry,the reports of bovine enterovirus infection have gradually increased.As a major animal husbandry province in China,Xinjiang focuses on the economic development of agriculture and animal husbandry.In recent years,with the rapid development of cattle breeding industry,epidemic diseases characterized by digestive system and respiratory system have occurred in cattle in many areas,causing serious economic losses to the cattle breeding industry in this area.In order to determine whether enterovirus infection exists in Xinjiang,we used double antibody sandwich ELISA to detect BEV antigen in 740 fecal samples collected from cattle herds in different regions of Xinjiang,and found that the BEV infection rate of cattle in different areas of Xinjiang was 7.8% ～ 24.3%,and the infection rate in southern Xinjiang was the highest,especially in Atush,reaching 24.3%.In order to determine the pathogenic species and characteristics of diarrhea in cattle in Xinjiang,we used cell culture technology to isolate and identify BEV-positive samples ELISA.Biological characteristics,physicochemical properties and genetic variations were characterized for the isolated strains.In addition,RT-PCR was used to amplify the 5’UTR genes of BEV isolates strains.The results showed that the Eighteen strains of BEV were successfully isolated.After inoculated to Vero cells,the isolated strains caused regular cell lesions such as shrinkage and rounding,enhanced refraction,disintegration and death.Characterization of physicochemical,biological and immunological characteristics of the isolated virus strains showed that the 18 virus strains had certain acid resistance,were insensitive to chloroform and ether,and were sensitive to high temperature.In order to further determine the molecular differences of bovine enterovirus(BEV)strains isolated from different regions of Xinjiang,RT-PCR was used to amplify the VP1 genes of BEV isolates,and their sequences were determined and compared.Analysis of the VP1 sequences showed that the 18 strains were all E enterovirus(EVE)and shared sequence identity of 71.4%～95.4% to the first domestic EV-E strain HY12.The homology between the isolated strains was 76.4%～98.4%.The homology between the newly isolated XJZP-29 and the foreign standard reference strain LC-R4 was only 61.2%.Analysis of the glycosylation sites,amino acid sites and secondary structures for the 18 virus strains using Bioinformatics software revealed unique glycosylation sites.in some strains of 18 virus strains.Phylogenetic tree analysis showed that the 18 EV-E strains could be further divided into three genotypes: EV-E1,EV-E2 and EV-E3.In summary,we have reveal the BEV infection in cattle herds in different regions of Xinjiang with a different infection rate.All 18 isolated enterovirus strains belongs to EV-E and could further divided into EV-E1,EV-E2 and EV-E3,which enriched the epidemiological data of BEV infection in China,and provided a theoretical basis for the prevention and control of the disease in Xinjiang in the future. |
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