石河子地区宠物犬蜱的种类鉴定及其携带布鲁氏菌的检测

为了解新疆石河子地区宠物犬体表寄生蜱的种类及蜱携带布鲁氏菌情况，本试验在形态学分类的基础上，基于线粒体基因12S rRNA和细胞色素氧化酶Ⅰ（COⅠ）对宠物犬体表采集寄生蜱进行分子生物学检测，使用DNAMAN软件比对分析序列同源性，并应用序列分析软件Mega 7.0邻接法构建蜱种系统发育进化树，分析蜱种的遗传进化关系；基于布鲁氏菌外膜蛋白Omp22基因对宠物犬体表寄生蜱进行布鲁氏菌PCR检测，确定宠物犬蜱布鲁氏菌携带情况。结果显示，宠物犬体表寄生的436只蜱的形态学与线粒体基因（12S rRNA和COⅠ）分子生物学鉴定结果一致，均为新疆优势蜱种之一——图兰扇头蜱（Rhipicephalus turanicus）。基于12S rRNA基因构建的蜱种系统发育树显示，本试验所得宠物犬体表寄生图兰扇头蜱序列与GenBank中已知图兰扇头蜱的序列聚为一大支。基于布鲁氏菌Omp22基因的巢式PCR扩增结果显示，宠物犬体表寄生图兰扇头蜱携带布鲁氏菌的阳性率为4.82%（21/436），且同源性为100%。BLAST比对显示，本试验所得宠物犬图兰扇头蜱携带的布鲁氏菌与新疆流行株YC31（GenBank登录号：MK201679.1）、QH5（GenBank登录号：MK201678.1）、EM3（GenBank登录号：MK201677.1）和ML9（GenBank登录号：MK201676.1）的同源性均为100%。本研究通过形态学及分子生物学探讨了新疆石河子地区宠物犬体表寄生蜱的种类及携带布鲁氏菌基本情况，为宠物犬体表寄生蜱的种类及蜱传疾病的监测和控制等研究工作提供了基础资料。

Species Identification of Ticks from Pet Dog and Detection of Brucella Carried by Ticks in Shihezi Area

To understand the species of ticks from pet dogs and their carrying status of Brucella in Shihezi,Xinjiang,on the basis of morphological classification,the ticks from pet dogs were detected by molecular biology method based on mitochondrial gene 12S rRNA and cytochrome oxidase Ⅰ (COⅠ).DNAMAN software was used to compare and analyze sequence homology,and sequence analysis software Mega 7.0 was used to construct a phylogenetic tree of tick species by using the Neighbor Joining method to analyze the genetic evolutionary relationship of tick species.Based on the Brucella outer membrane protein Omp22 gene,ticks from pet dogs were tested for Brucella by PCR to determine the Brucella carrying status of ticks from pet dogs.The results showed that the morphology of 436 ticks from pet dogs were consistent with the molecular biology identification results of mitochondrial genes (12S rRNA and COⅠ),and they were all Rhipicephalus turanicus,one of the dominant species in Xinjiang.The phylogenetic tree of tick species constructed based on the 12S rRNA gene showed that the sequence of Rhipicephalus turanicus from pet dogs obtained in this experiment was clustered well with the sequence of the corresponding tick known in GenBank.The results of nested PCR amplification based on the Omp22 gene of Brucella showed that the positive rate of Brucella carried by Rhipicephalus turanicus from pet dogs was 4.82% (21/436),and the homology was 100%.BLAST comparison showed that Brucella carried by Rhipicephalus turanicus from pet dogs obtained in this experiment was related to the Xinjiang epidemic strain YC31 (GenBank accession No.:MK201679.1),QH5 (GenBank accession No.:MK201678.1),EM3 (GenBank accession No.:MK201677.1) and ML9 (GenBank accession No.:MK201676.1),and the homologies were all 100%.In this study,the species of ticks from pet dogs in Shihezi,Xinjiang,and their carrying Brucella were studied by morphology and molecular biology,and provided basic information for the species of parasitic ticks on pet dogs and the monitoring and control of tick-borne diseases.