新疆喀什地区安格斯犊牛腹泻主要病毒性病原调查及牛诺如病毒全基因序列分析

doi:10.16431/j.cnki.1671-7236.2022.11.033

中国畜牧兽医 ›› 2022, Vol. 49 ›› Issue (11): 4420-4428.doi: 10.16431/j.cnki.1671-7236.2022.11.033

新疆喀什地区安格斯犊牛腹泻主要病毒性病原调查及牛诺如病毒全基因序列分析

吴静¹, 史晋成¹, 萨妮耶·库尔班¹, 纳曼·阿卜杜拉¹, 孙磊¹, 姚刚¹, 钟旗², 骆梁涛⁴, 赵红琼¹, 马雪连^1,3

1. 新疆农业大学动物医学学院, 乌鲁木齐 830052;
2. 新疆畜牧科学院兽医研究所, 乌鲁木齐 830052;
3. 新疆农业大学畜牧学博士后流动站, 乌鲁木齐 830052;
4. 新疆刀郎阳光农牧科技股份有限公司, 麦盖提 844600

收稿日期:2022-05-29 出版日期:2022-11-05 发布日期:2022-11-04
通讯作者: 赵红琼, 马雪连 E-mail:zhaohongqiong@sina.com;13699381790@163.com
作者简介:吴静,E-mail:1601056628@qq.com。
基金资助:
科技厅乡村振兴项目（202113563）；肉牛高效健康饲养技术集成研究与示范（2021A02003-1）；自治区天山青年基金（2020Q075）

Investigation of the Main Viral Pathogens of Angus Calf Diarrhea in Kashgar Region of Xinjiang and Whole Gene Sequence Analysis of Bovine Norovirus

WU Jing¹, SHI Jincheng¹, SANIYE Kerban¹, NAMAN Abudula¹, SUN Lei¹, YAO Gang¹, ZHONG Qi², LUO Liangtao⁴, ZHAO Hongqiong¹, MA Xuelian^1,3

1. College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, China;
2. Institute of Veterinary Medicine, Xinjiang Academy of Animal Sciences, Urumqi 830052, China;
3. Postdoctoral Mobile Station of Animal Husbandry, Xinjiang Agricultural University, Urumqi 830052, China;
4. Xinjiang Daolang Sunshine Agriculture and Animal Husbandry Technology Corporation, McGetty 844600, China

Received:2022-05-29 Online:2022-11-05 Published:2022-11-04

摘要/Abstract

摘要： 【目的】调查新疆喀什地区安格斯犊牛腹泻主要病毒性病原流行情况及牛诺如病毒（Bovine norovirus，BNoV）全基因组遗传关系。【方法】采用RT-PCR技术分别对2021年不同季节在喀什4个牛场采集的363份犊牛腹泻粪便样品进行牛轮状病毒（Bovine rotavirus，BRV）、牛冠状病毒（Bovine coronavirus，BCoV）、牛病毒性腹泻病毒（Bovine viral diarrhea virus，BVDV）和BNoV 4种病毒性病原检测，并对扩增获得的BNoV全基因序列进行分析。【结果】 BCoV和BNoV检出率较高，分别为36.09%（131/363）和25.07%（91/363），混合感染中BCoV＋BNoV检出数量最多，阳性率为44.00%（22/50）。对喀什地区4个场区进行检测，其中A场主要检测出BCoV和BNoV，检出率分别为43.52%（47/108）和32.41%（35/108）；B场主要检测到BCoV，检出率为52.21%（71/136）；C、D场病原检出率普遍较低。秋、冬季检出量最多的病毒分别为BNoV和BCoV，检出率分别为66.67%（50/75）和77.59%（90/116）。对BNoV全基因序列分析显示，本试验检测到的新疆喀什地区BNoV Bo/XJ-KS/01/CHN株（登录号：ON076888）属于GⅢ.2型BNoV，与中国CN/HB-SJZ和CH/HB/BD/2019毒株在进化树中处于同一分支，且与CN/HB-SJZ-2毒株核苷酸相似性最高，为93.70%；与英国分离到的Jena/1999/UK毒株核苷酸相似性最低，为72.49%。进一步对3个开放阅读框（ORF）比对分析发现，测得的BNoV全基因组序列与国内参考株CN/HB-SJZ-2的核苷酸和氨基酸相似性主要是在ORF1区域最高，分别为94.24%和98.69%；与国外参考株Jena/1999/UK的核苷酸和氨基酸相似性主要是在ORF3区域最低，分别为63.59%和64.57%。与国内外毒株比对结果显示，未出现基因重组现象。【结论】南疆地区犊牛腹泻流行情况因季节、场区不同有一定的差异。病毒性病原主要在秋、冬季感染率较高，以单一BCoV、BRV和BNoV感染为主。本研究测定分析的BNoV GⅢ.2型Bo/XJ-KS/01/CHN全基因序列与中国CN/HB-SJZ-2参考株亲缘关系最近，与国内外全基因组参考序列比对未出现基因重组现象。

关键词: 犊牛腹泻; 病毒性病原; 流行病学调查; 遗传进化分析

Abstract: 【Objective】 The aim of this experiment was to investigate the prevalence of the main viral pathogens of Angus calves diarrhea in Kashgar, Xinjiang and the genome-wide genetic relationship of Bovine norovirus (BNoV).【Method】 RT-PCR was used to detect four viral pathogens of Bovine rotavirus (BRV), Bovine coronavirus (BCoV), Bovine viral diarrhoea virus (BVDV) and BNoV in 363 calf diarrhea fecal samples collected from 4 cattle farms in Kashgar in different seasons in 2021, and the amplified BNoV gene sequences were analyzed.【Result】 The positive rates of BCoV and BNoV were 36.09% (131/363) and 25.07% (91/363), respectively.In mixed infection, BCoV+BNoV was the most, with a positive rate of 44.00% (22/50).Four fields in Kashgar area were tested.BCoV and BNoV were mainly detected in field A, with a detection rate of 43.52% (47/108) and 32.41% (35/108), respectively.BCoV was mainly detected in field B, with a detection rate of 52.21% (71/136).The pathogen detection rates in fields C and D were generally low.The viruses detected the most in autumn and winter were BNoV and BCoV, with detection rates of 66.67% (50/75) and 77.59% (90/116), respectively.The analysis of the whole gene sequence of BNoV showed that BNoV strain Bo/XJ-KS/01/CHN (accession No.:ON076888) detected in this experiment belonged to BNoV GⅢ.2, and was in the same branch of the evolutionary tree as CN/HB-SJZ-2 and CH/HB/BD/2019 strains in China, and had the highest nucleotide similarities with CN/HB-SJZ-2 strains, which was 93.70%.The nucleotide similarity with Jena/1999/UK strain isolated from Britain were the lowest, which was 72.49%. Further comparison and analysis of three open reading frames (ORFs) showed that the nucleotide and amino acid similarities between the whole genome sequence of BNoV and the domestic reference strain CN/HB-SJZ-2 were mainly the highest in ORF1 region, with 94.24% and 98.69%, respectively.The nucleotide and amino acid similarities with the foreign reference strain Jena/1999/UK was the lowest in ORF3 region, which was 63.59% and 64.57%, respectively.The comparison with domestic and foreign strains showed that there was no gene recombination.【Conclusion】 The epidemic situation of calf diarrhea in Southern Xinjiang was different due to different seasons and fields.The infection rate of viral pathogens was mainly high in autumn and winter, and single BCoV, BRV and BNoV were the main infections.The whole gene sequence of BNoV type GⅢ.2 Bo/XJ-KS/01/CHN determined in this study had the closest relationship with the reference strain CN/HB-SJZ-2 in China, and had no gene recombination phenomenon compared with the whole genome reference sequence at home and abroad.

Key words: calf diarrhea; viral pathogens; epidemiological investigation; genetic evolutionary analysis

中图分类号:

S852.65^＋3

吴静, 史晋成, 萨妮耶·库尔班, 纳曼·阿卜杜拉, 孙磊, 姚刚, 钟旗, 骆梁涛, 赵红琼, 马雪连. 新疆喀什地区安格斯犊牛腹泻主要病毒性病原调查及牛诺如病毒全基因序列分析[J]. 中国畜牧兽医, 2022, 49(11): 4420-4428.

WU Jing, SHI Jincheng, SANIYE Kerban, NAMAN Abudula, SUN Lei, YAO Gang, ZHONG Qi, LUO Liangtao, ZHAO Hongqiong, MA Xuelian. Investigation of the Main Viral Pathogens of Angus Calf Diarrhea in Kashgar Region of Xinjiang and Whole Gene Sequence Analysis of Bovine Norovirus[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4420-4428.

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新疆喀什地区安格斯犊牛腹泻主要病毒性病原调查及牛诺如病毒全基因序列分析

Investigation of the Main Viral Pathogens of Angus Calf Diarrhea in Kashgar Region of Xinjiang and Whole Gene Sequence Analysis of Bovine Norovirus

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