Faculty of Veterinary Science, The University of Melbourne, Parkville, Victoria 3010, Australia.
Vet Parasitol. 2013 Nov 8;197(3-4):509-21. doi: 10.1016/j.vetpar.2013.06.023. Epub 2013 Jul 4.
This study investigated Theileria orientalis following outbreaks of oriental theileriosis in cattle in the state of Victoria, Australia, from September 2010 to January 2012, using traditional and molecular methods of diagnosis. A questionnaire was used to collect epidemiological information from cattle farms. Blood samples (n=301), collected from individual symptomatic and asymptomatic cattle from 19 cattle farms, were examined for the presence of Theileria on stained blood smears and tested using a PCR-based approach, employing a region within the major piroplasm surface protein (MPSP) gene as a marker. The microscopic examination of stained blood smears detected stages consistent with Theileria piroplasms in 28.1% (79/281) of the samples. PCR products were amplified from 70.8% (213/301) of the samples. Mutation scanning analysis of all amplicons displayed seven distinct profiles. Following the direct sequencing of representative amplicons, the genotypes ikeda, chitose, buffeli and type 5 were detected in 91.1%, 32.9%, 2.4% and 1.4% of 213 blood samples, respectively. The distribution of these four genotypes varied among the 19 farms; genotype ikeda was detected on all farms, whereas genotypes chitose, buffeli and type 5 were detected on 14, 3 and 2 farms, respectively. Mix infections with genotypes ikeda and chitose were common (21.6%). Survey results revealed that oriental theileriosis affected mainly beef cows of more than two years of age, prior to calving, and disease was associated with abortion and cow deaths. Future investigations should focus on developing improved tools for investigating and managing oriental theileriosis.
本研究调查了澳大利亚维多利亚州 2010 年 9 月至 2012 年 1 月间爆发的东方泰勒虫病,使用传统和分子诊断方法。采用问卷收集来自 19 个牛场的流行病学信息。从 19 个牛场的 19 头有症状和无症状牛采集血样(n=301),在染色血涂片上检查有无东方泰勒虫,并用基于 PCR 的方法进行检测,该方法使用主要血孢子虫表面蛋白(MPSP)基因内的一个区域作为标记。染色血涂片的显微镜检查在 28.1%(79/301)的样本中发现与泰勒虫属血孢子虫一致的阶段。从 70.8%(213/301)的样本中扩增出 PCR 产物。对所有扩增子进行突变扫描分析,显示出 7 种不同的图谱。对代表性扩增子进行直接测序后,在 91.1%(213/213)的血液样本中检测到 ikeda、chitose、buffeli 和 type 5 基因型,分别为 32.9%、2.4%和 1.4%。这四种基因型在 19 个牛场中的分布不同;ikeda 基因型在所有牛场中均有检出,而 chitose、buffeli 和 type 5 基因型分别在 14、3 和 2 个牛场中检出。ikeda 和 chitose 基因型的混合感染很常见(21.6%)。调查结果表明,东方泰勒虫病主要影响产犊前 2 岁以上的肉牛,与流产和奶牛死亡有关。未来的研究应重点开发改进的工具来调查和管理东方泰勒虫病。