| 新疆南部图兰扇头蜱及卵携带R.raoultii的分子检测 |
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| 引用本文： | 刘永宏,李贝贝,李凯瑞,何波,张敬博,蒲小峰,陈梅娟,潘姣姣,李飞,张路瑶,赵丽.新疆南部图兰扇头蜱及卵携带R.raoultii的分子检测[J].中国农业科学,2018,51(15):3020-3028. |
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| 作者姓名： | 刘永宏 李贝贝 李凯瑞 何波 张敬博 蒲小峰 陈梅娟 潘姣姣 李飞 张路瑶 赵丽 |
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| 作者单位： | 塔里木大学动物科学学院/新疆生产建设兵团塔里木畜牧科技重点实验室;新疆阿克苏地区动物疫病控制诊断中心;巴里坤哈萨克自治县畜牧兽医工作站 |
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| 基金项目： | 国家自然科学基金（31460655）、大学生创新创业训练计划（107572017009，107572017014）、新疆生产建设兵团塔里木畜牧科技重点实验室开放课题（HS201501，HS201801） |
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| 摘 要： | 【目的】世界上蜱类3科18属899种,中国有2科10属117种,新疆至少有2科10属45种,占全国蜱种类的1/3之多,分布也极其广泛。蜱直接危害和传播多种病原,且一些病原可经卵垂直传播,给畜牧业造成巨大经济损失,还严重威胁公共卫生安全。图兰扇头蜱是新疆南部荒漠及半荒漠地区常见种和优势种。确定新疆南部图兰扇头蜱及其卵是否携带立克次体,对该蜱及其传播立克次体病的防控意义重大。【方法】对新疆生产建设兵团塔里木畜牧科技重点实验室在新疆南部阿拉尔市某羊场收集的饱血雌性扇头蜱,置于一定湿度和一定温度的环境中产卵,随机取5个独立的卵样品及其对应的5只雌蜱为研究对象。通过对雌蜱及其卵分别处理后提取基因组DNA,进行PCR扩增蜱12S r RNA基因和立克次体16S r RNA基因,并对扩增产物测序,利用BLAST在线平台和多个分子生物学软件进行序列分析。【结果】5只雌蜱12S r RNA基因PCR扩增全部阳性,测序获得的4段蜱12S r RNA基因序列完全一致;Blast分析与Gen Bank数据库中图兰扇头蜱12S r RNA基因序列相似性高达99%以上,且高相似性前五基因序列均为图兰扇头蜱,其中包括来自新疆绵羊的图兰扇头蜱;本研究蜱12S r RNA基因序列提交Gen Bank数据库获得登录号为MG744514,与来自于Gen Bank数据库图兰扇头蜱、血红扇头蜱、微小牛蜱、边缘革蜱、草原革蜱、长角血蜱、小亚璃眼蜱、亚洲璃眼蜱、残缘璃眼蜱、全沟硬蜱及外围群尘螨的22个12S r RNA基因序列的进化树显示,研究所获得的蜱12S r RNA基因序列与图兰扇头蜱进化关系最近,聚在同一个小分支;确定了该扇头蜱为图兰扇头蜱。5只产卵后的雌蜱和相应蜱所产的全部卵立克次体16S r RNA基因PCR扩增,有1只蜱和其产的卵样品阳性,蜱携带率为20%;雌蜱及其卵立克次体16S r RNA基因测序结果完全一致;Blast分析与Gen Bank数据库中Rickettsia raoultii 16S r RNA基因序列相似性高达99%以上,且高相似性前五基因序列为4个Rickettsia raoultii和1个Rickettsia sp.,其中包括来自新疆2011年的亚洲璃眼蜱和草原革蜱的立克次体;立克次体16S r RNA基因序列提交Gen Bank数据库获得登录号为MG744513,与来自于Gen Bank数据库的37个24种立克次体16S r RNA基因序列的进化树显示,研究获得的立克次体16S r RNA基因序列与Rickettsia raoultii进化关系最近,聚在同一个小分支,与其他15种立克次体同属于斑点热群立克次体;确定了本研究图兰扇头蜱及其卵均携带斑点热群立克次体R.raoultii。【结论】首次发现图兰扇头蜱及其卵携带R.raoultii。
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| 关 键 词： | 劳氏立克次体 卵 图兰扇头蜱 新疆 |
| 收稿时间： | 2018-01-05 |
Molecular Detection of R. turanicus and Its Eggs Carrying R. raoultii in Southern Xinjiang |
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| LIU YongHong,LI BeiBei,LI KaiRui,HE Bo,ZHANG JingBo,PU XiaoFeng,CHEN MeiJuan,PAN JiaoJiao,LI Fei,ZHANG LuYao,ZHAO Li.Molecular Detection of R. turanicus and Its Eggs Carrying R. raoultii in Southern Xinjiang[J].Scientia Agricultura Sinica,2018,51(15):3020-3028. |
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| Authors: | LIU YongHong LI BeiBei LI KaiRui HE Bo ZHANG JingBo PU XiaoFeng CHEN MeiJuan PAN JiaoJiao LI Fei ZHANG LuYao ZHAO Li |
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| Institution: | 1.College of Animal Science, Tarim University/Key Laboratory of Tarim Animal Husbandry Science and Technology of Xinjiang Production & Construction Corps, Alar 843300, Xinjiang;2.Animal Loimia Controlling and Diagnostic Center of Aksu Region, Aksu 843000, Xinjiang;3.Animal husbandry and veterinary workstations of Barkol kazak autonomous county, Kumul 839200, Xinjiang |
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| Abstract: | 【Objective】 There are 899 species of ticks in the world, belonging to 18 genera and 3 families, and there are 117 species in China, belonging to 2 families, 10 genera. There are at least 45 species in Xinjiang, accounting for more than 1/3 of the national ticks, and its distribution is extremely wide. Ticks can directly harm and spread many kinds of pathogens, and some pathogens can propagate vertically through eggs, causing huge economic losses to animal husbandry, and seriously threatening public health security. Rhipicephalus is one of the common species and dominant species in the desert and semi-desert region of southern Xinjiang. It is of great significance to confirm whether Rhipicephalus turanicus and its eggs are carrying the Rickettsia in the southern Xinjiang of China, so as to prevent and control of Rhipicephalus turanicus and its transmission of Rickettsia. 【Method】 The satiated blood female Rhipicephalus were collected from a sheep farm in alar, southern Xinjiang, and oviposited eggs at a certain humidity and temperature. A random sample of 5 independent eggs and the corresponding 5 female Rhipicephalus were as the research object. The genomic DNA were extracted from the female Rhipicephalus and eggs, respectively. In this study, ticks 12S rRNA gene and Rickettsia 16S rRNA gene were amplified by conventional PCR, and the amplification products were sequenced and underwent sequence analysis by using BLAST online platform and multiple molecular biology software. 【Result】 The 12S rRNA gene PCR amplification of 5 female ticks were all positive, and 4 segment of 12S rRNA gene sequence obtained by sequencing were identical. The similarity of Rhipicephalus 12s rRNA gene sequence and GenBank Rhipicephalus 12s rRNA gene sequences in the database were more than 99% using Blast analysis, and the top five gene sequences of the high similarity were Rhipicephalus, including Rhipicephalus from Xinjiang sheep. In this study, the Rhipicephalus 12S rRNA gene sequence was submitted to the GenBank database and obtained the login number of MG744514. The phylogenetic tree from GenBank database Rhipicephalus turanicus, Rhipicephalus sanguineus, Boophilus microplus, Dermacentor marginatus, Dermacentor nuttalli, Haemaphysalis longicornis, Hyalomma anatolicum, Hyalomma asiaticum, Hyalomma detritum, Ixodes persulcatus and peripheral-dust mites 12S rRNA gene sequence display, the Rhipicephalus 12S rRNA gene sequence obtained in this study was closest with Rhipicephalus sanguineus in the evolutionary relationship and clustered in the same small branch. Rhipicephalus was identified as Rhipicephalus turanicus. PCR amplification of all eggs Rickettsia 16S rRNA gene and 5 oviposit female Rhipicephalus, 1 Rhipicephalus and its eggs samples were positive, and the tick carrying rate was 20%. The sequencing results of 16S rRNA in female Rhipicephalus turanicus and its eggs were identical. The similarity was more than 99% of the Rickettsia raoultii 16S rRNA gene sequences in the GenBank database using Blast analysis. The top five genes of high similarity were 4 Rickettsia raoultii and 1 Rickettsia sp, including the rickettsia of the Hyalomma asiaticum and Dermacentor nuttalli from Xinjiang in 2011. In this study, the Rickettsia 16S rRNA gene sequence was submitted to the GenBank database and obtained the login number of MG744513. Evolutionary tree of 37 Rickettsia 16S rRNA gene sequences belonging to 24 species from the GenBank database showed, the Rickettsia 16S rRNA gene sequence obtained in this study was closest with Rickettsia raoultii in the evolutionary relationship and clustered in the same small branch, belonging to the speckle thermal gro up rickettsia with the other 15 kinds of rickettsia. Rhipicephalus turanicus and its eggs carried Rickettsia raoultii of spotted fever group Rickettsia. 【Conclusion】 R. raoultii were first found in Rhipicephalus turanicus and its eggs. |
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| Keywords: | Rickettsia raoultii eggs Rhipicephalus turanicus Xinjiang |
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