畜牧与饲料科学 ›› 2024, Vol. 45 ›› Issue (4): 109-114.doi: 10.12160/j.issn.1672-5190.2024.04.014

• 动物疾病防控 • 上一篇    下一篇

新疆生产建设兵团第九师肉牛感染泰勒原虫的PCR检测与种类鉴定

朱婷婷1,2, 曹梦雅2, 司俊飞2, 张振杰2, 张旭2, 苏政1, 刘鹏1, 彭健1, 齐萌2   

  1. 1.新疆生产建设兵团第九师农业科学研究所(畜牧科学研究所),新疆 额敏 834601;
    2.塔里木大学动物科学与技术学院,新疆 阿拉尔 843300
  • 收稿日期:2024-04-28 出版日期:2024-07-30 发布日期:2024-08-23
  • 通讯作者: 齐萌(1985—),男,教授,博士,硕士生导师,主要从事人兽共患病病原生物学研究工作。
  • 作者简介:朱婷婷(1985—),女,硕士研究生,主要研究方向为动物群发病防控。
  • 基金资助:
    新疆生产建设兵团第九师科技计划项目(2021JS001,2022JS008,2023JS002)

PCR Detection and Species Identification of Theileria in Beef Cattle in the Ninth Division of the Xinjiang Production and Construction Corps

ZHU Tingting1,2, CAO Mengya2, SI Junfei2, ZHANG Zhenjie2, ZHANG Xu2, SU Zheng1, LIU Peng1, PENG Jian1, QI Meng2   

  1. 1. Agricultural Science Research Institute (Animal Husbandry Science Research Institute)of the Ninth Division of the Xinjiang Production and Construction Corps,Emin 834601,China;
    2. College of Animal Science and Technology,Tarim University,Alar 843300,China
  • Received:2024-04-28 Online:2024-07-30 Published:2024-08-23

摘要: [目的]掌握新疆生产建设兵团第九师(以下称第九师)肉牛泰勒原虫的感染情况。[方法]从第九师161团、163团、164团和165团规模化肉牛养殖场或养殖户,分别采集肉牛抗凝血48、22、25、23份,其中,规模化养殖肉牛48份,散养肉牛70份,提取血液全基因组DNA。利用设计的特异性引物,采用PCR法对肉牛全血DNA样本分别进行环形泰勒虫、东方泰勒虫、中华泰勒虫检测,通过序列比对鉴定泰勒原虫种类,构建系统发育树进行种类遗传进化分析。采用χ2检验法分析不同采样地点、不同饲养模式下肉牛泰勒原虫的感染率差异。[结果]PCR检测结果显示,在118份肉牛血液DNA样本中,检测出泰勒原虫阳性样本82份,感染率为69.49%(82/118);检测到的阳性样本均为东方泰勒虫,未检测到环形泰勒虫和中华泰勒虫。第九师161团、163团、164团和165团肉牛的泰勒原虫感染率分别为29.17%(14/48)、95.45%(21/22)、96.00%(24/25)和100%(23/23);不同采样地点肉牛泰勒原虫的感染率统计学差异极显著(χ2=62.195,P<0.01)。散养条件下肉牛泰勒原虫感染率较高,为97.14%(68/70),规模化养殖条件下肉牛泰勒虫感染率较低,为29.17%(14/48);不同饲养模式下肉牛泰勒原虫的感染率差异极显著(χ2=62.061,P<0.01)。在获得的82条序列中,有78条与我国新疆维吾尔自治区牛源东方泰勒虫MPSP基因序列(GenBank登录号:ON462019)同源性为100%,序列型命名为TSE1;有4条与我国新疆维吾尔自治区牛源东方泰勒虫MPSP基因序列(GenBank登录号:ON462018)同源性为100%,序列型命名为TSE2。种系发育分析结果显示,鉴定的东方泰勒虫TSE1和TSE2序列均与我国新疆维吾尔自治区牛源东方泰勒虫聚成一支。[结论]新疆生产建设兵团第九师肉牛感染的泰勒原虫种类为东方泰勒虫,散养条件下感染率更高,应加强对散养肉牛泰勒原虫的检测。

关键词: 肉牛, 泰勒原虫, 检测, 鉴定

Abstract: [Objective] The aim of the present study was to clarify the infection status of Theileria in beef cattle in the Ninth Division of the Xinjiang Production and Construction Corps (hereinafter referred to as the Ninth Division). [Method] A total of 118 blood samples were collected with anticoagulation tubes from beef cattle reared in the large-scale farms or scatter-feed households in the 161 (n=48), 163 (n=22), 164 (n=25) and 165 (n=23) Regiments of the Ninth Division, respectively. Among them, 48 samples were collected from the confined beef cattle and the remaining 70 samples were collected from the free-range beef cattle. The whole genomic DNA of the blood samples were extracted, and PCR assay was used to detect the presence of T. annulata, T. orientalis and T. sinensis with the specially designed primers. The species of Theileria were identified by sequence alignment, and the phylogenetic tree was constructed for genetic evolution analysis. The χ2 test was employed to assess the differences in the infection rate of Theileria in beef cattle at different sampling sites and under different rearing modes. [Result] PCR detection showed that among the 118 blood DNA samples of beef cattle, 82 samples were positive for Theileria, with an overall infection rate of 69.49% (82/118). All Theileria positive samples had T. orientalis alone, while T. annulata and T. sinensis were absent. The infection rates of Theileria in beef cattle in the 161, 163, 164 and 165 Regiments of the Ninth Division were 29.17% (14/48), 95.45% (21/22), 96.00% (24/25) and 100% (23/23), respectively. There was an extremely significant difference in the infection rates of Theileria in beef cattle at different sampling sites (χ2=62.195, P<0.01). A higher infection rate (97.14%, 68/70) of Theileria was observed in the free-range beef cattle, while a lower one (29.17%, 14/48) was observed in the confined beef cattle. The infection rates of Theileria in beef cattle under different rearing modes were extremely significantly different (χ2=62.061, P<0.01). Among the 82 sequences obtained, 78 had 100% homology with the MPSP gene sequence of a T. orientalis isolate from cattle in Xinjiang Uygur Autonomous Region (GenBank accession number: ON462019), and the sequence type was designated as TSE1. The remaining 4 sequences had 100% homology with the MPSP gene sequence of another T. orientalis strain of cattle origin in Xinjiang Uygur Autonomous Region (GenBank accession number: ON462018), and the sequence type was designated as TSE2. The phylogenetic analysis demonstrated that the identified TSE1 and TSE2 sequences in present study were clustered with T. orientalis of cattle origin in Xinjiang Uygur Autonomous Region. [Conclusion] The species of Theileria prevailed in beef cattle in the Ninth Division of the Xinjiang Production and Construction Corps was T. orientalis. The free-range beef cattle had a higher infection rate of Theileria, suggesting the detection of Theileria in the free-range beef cattle herd should be strengthened.

Key words: beef cattle, Theileria, detection, identification

中图分类号: