西藏牦牛、绵羊棘球蚴病病原的分子鉴定

doi:10.16431/j.cnki.1671-7236.2020.04.027

中国畜牧兽医 ›› 2020, Vol. 47 ›› Issue (4): 1209-1215.doi: 10.16431/j.cnki.1671-7236.2020.04.027

西藏牦牛、绵羊棘球蚴病病原的分子鉴定

夏晨阳¹, 石斌¹, 唐文强¹, 单曲拉姆¹, 贾万忠², 白吉³, 吴燕涛², 闫鸿斌², 刘建枝¹

1. 西藏自治区农牧科学院畜牧兽医研究所, 拉萨 850009;
2. 中国农业科学院兰州兽医研究所, 家畜疫病病原生物学国家重点实验室, 甘肃省动物寄生虫病重点实验室, 兰州 730046;
3. 西藏自治区动物疫病预防控制中心, 拉萨 850007

收稿日期:2019-08-19 出版日期:2020-04-20 发布日期:2020-04-17
通讯作者: 闫鸿斌, 刘建枝 E-mail:yanhongbin@caas.cn;13659526886@163.com
作者简介:夏晨阳(1980-),男,黑龙江明水人,硕士,研究方向,家畜寄生虫学,E-mail:13518978860@163.com
基金资助:
牛羊内外寄生虫病调查及防控关键技术研究与示范（XZ201901NA02）；家畜疫病病原生物学国家重点实验室（中国农业科学院兰州兽医研究所）开放课题（SKLVEB2016KFKT011）；西藏自治区科技厅重点计划项目——西藏牛羊包虫病、皮蝇蛆病防控技术研究与示范；国家公益性行业（农业）科研专项（201303037）

Molecular Identification of Hydatid Cysts from Sheep and Yaks of Tibet

XIA Chenyang¹, SHI Bin¹, TANG Wenqiang¹, Danqulamu¹, JIA Wanzhong², Bai Ji³, WU Yantao², YAN Hongbin², LIU Jianzhi¹

1. Institute of Animal Science, Tibet Academy of Agricultural and Animal Husbandry Sciences, Lhasa 850009, China;
2. State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;
3. Center for Animal Disease Control and Prevention of Tibet, Lhasa 850007, China

Received:2019-08-19 Online:2020-04-20 Published:2020-04-17

摘要/Abstract

摘要： 本研究旨在对西藏自治区那曲地区和拉萨市牦牛、绵羊体内棘球蚴病原进行分子生物学鉴定并分析其遗传变异规律。对2016年11月底采自西藏拉萨市当雄县和那曲地区嘉黎县的5只绵羊体内的5个棘球蚴包囊、15头牦牛体内的18个棘球蚴包囊分别分离棘球蚴原头蚴或生发层组织，提取基因组DNA，应用PCR方法扩增nad1基因，通过测序获得nad1全基因序列。运用DNAStar MegAlign软件对序列进行同源性分析。以GenBank中已公布的棘球属的nad1全基因序列为比对对象，采用最大似然法（ML）构建系统发育树。结果显示，所测定的牦牛和绵羊的23个棘球蚴病原nad1基因序列与GenBank登录的细粒棘球蚴狭义种（G1基因型）nad1基因序列高度同源，同源性为99.6%~99.8%，23条nad1基因的遗传距离为0~0.0022447。同源基因的碱基变异率为0.2%~0.4%；与棘球属其他棘球绦虫同源基因的碱基变异率为14.9%~19.8%。有5个样本的nad1基因在不同位点发生碱基突变，变异位点发生序列转换。以上结果表明，本研究所采集牦牛和绵羊的棘球蚴为细粒棘球绦虫G1基因型，其nad1基因变异小，序列一致性高。

关键词: 西藏; 牦牛; 绵羊; 棘球蚴; 分子鉴定

Abstract: The purpose of this study was to identify the hydatid pathogen in yaks and sheep in Naqu and Lhasa of Tibet Autonomous Region and analyze its genetic variation.5 hydatid cysts were isolated from 5 sheep and 18 hydatid cysts from 15 yaks collected in Dangxiong county of Lhasa,and Jiali county of Naqu at the end of November 2016.Genomic DNA was extracted,nad1 gene was amplified by PCR,and nad1 full gene sequence was obtained by sequencing.DNAStar MegAlign software was used to analyze the sequence homology.Taking nad1 gene sequence of Echinococcus published in GenBank as comparison object,the phylogenetic tree was constructed by ML.The results showed that the nad1 gene sequence of 23 hydatid pathogens from yaks and sheep was highly homologous with that of the gene sequence of Echinococcus granulosus (G1 genotype),the homology was 99.6% to 99.8%,and the genetic distance of 23 nad1 genes was 0 to 0.0022447.The base variation rate of homologous genes was 0.2% to 0.4%,and that of homologous genes of Echinococcus was 14.9% to 19.8%.5 samples of nad1 gene had base mutation at different sites,and the mutation sites had sequence conversion.The results showed that the Echinococcus of yaks and sheep collected in this study were G1 genotype of Echinococcus granulosus,its nad1 gene had small variation and high sequence consistency.

Key words: Tibet; yak; sheep; hydatid cyst; molecular classification

中图分类号:

S855.9

夏晨阳, 石斌, 唐文强, 单曲拉姆, 贾万忠, 白吉, 吴燕涛, 闫鸿斌, 刘建枝. 西藏牦牛、绵羊棘球蚴病病原的分子鉴定[J]. 中国畜牧兽医, 2020, 47(4): 1209-1215.

XIA Chenyang, SHI Bin, TANG Wenqiang, Danqulamu, JIA Wanzhong, Bai Ji, WU Yantao, YAN Hongbin, LIU Jianzhi. Molecular Identification of Hydatid Cysts from Sheep and Yaks of Tibet[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(4): 1209-1215.

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西藏牦牛、绵羊棘球蚴病病原的分子鉴定

Molecular Identification of Hydatid Cysts from Sheep and Yaks of Tibet

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