土耳其双芽巴贝斯虫、环形泰勒虫、东方泰勒虫和边缘无形体的分子检测与基因鉴定
Molecular detection and genetic identification of Babesia bigemina, Theileria annulata, Theileria orientalis and Anaplasma marginale in Turkey.
作者信息
Zhou Mo, Cao Shinuo, Sevinc Ferda, Sevinc Mutlu, Ceylan Onur, Moumouni Paul Franck Adjou, Jirapattharasate Charoonluk, Liu Mingming, Wang Guanbo, Iguchi Aiko, Vudriko Patrick, Suzuki Hiroshi, Xuan Xuenan
机构信息
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Faculty of Veterinary Medicine, Selcuk University, Konya, Turkey.
出版信息
Ticks Tick Borne Dis. 2016 Feb;7(1):126-134. doi: 10.1016/j.ttbdis.2015.09.008. Epub 2015 Sep 28.
Babesia spp., Theileria spp. and Anaplasma spp. are significant tick-borne pathogens of livestock globally. In this study, we investigated the presence and distribution of Babesia bigemina, Theileria annulata, Theileria orientalis and Anaplasma marginale in cattle from 6 provinces of Turkey using species-specific PCR assays. The PCR were conducted using the primers based on the B. bigemina rhoptry-associated protein 1a (BbiRAP-1a), T. annulata merozoite surface antigen-1 (Tams-1), T. orientalis major piroplasm surface protein (ToMPSP) and A. marginale major surface protein 4 (AmMSP4) genes, respectively. Fragments of B. bigemina internal transcribed spacer (BbiITS), T. annulata internal transcribed spacer (TaITS), ToMPSP and AmMSP4 genes were sequenced for phylogenetic analysis. PCR results revealed that the overall infections of A. marginale, T. annulata, B. bigemina and T. orientalis were 29.1%, 18.9%, 11.2% and 5.6%, respectively. The co-infection of two or three pathogens was detected in 29/196 (15.1%) of the cattle samples. The results of sequence analysis indicated that BbiRAP-1a, BbiITS, Tams-1, ToMPSP and AmMSP4 were conserved among the Turkish samples, with 99.76%, 99-99.8%, 99.34-99.78%, 96.9-99.61% and 99.42-99.71% sequence identity values, respectively. In contrast, the Turkish TaITS gene sequences were relatively diverse with 92.3-96.63% identity values. B. bigemina isolates from Turkey were found in the same clade as the isolates from other countries in phylogenetic analysis. On the other hand, phylogenetic analysis based on T. annulata ITS sequences revealed significant differences in the genotypes of T. annulata isolates from Turkey. Additionally, the T. orientalis isolates from Turkish samples were classified as MPSP type 3 genotype. This is the first report of type 3 MPSP in Turkey. Moreover, AmMSP4 isolates from Turkey were found in the same clade as the isolates from other countries. This study provides important data for understanding the epidemiology of tick-borne diseases and it is expected to improve approach for diagnosis and control of tick-borne diseases in Turkey.
巴贝斯虫属、泰勒虫属和无形体属是全球范围内重要的蜱传家畜病原体。在本研究中，我们使用种特异性PCR检测方法，调查了土耳其6个省份牛体内双芽巴贝斯虫、环形泰勒虫、东方泰勒虫和边缘无形体的存在情况与分布。PCR分别使用基于双芽巴贝斯虫棒状体相关蛋白1a（BbiRAP - 1a）、环形泰勒虫裂殖子表面抗原-1（Tams - 1）、东方泰勒虫主要梨形虫表面蛋白（ToMPSP）和边缘无形体主要表面蛋白4（AmMSP4）基因的引物进行。对双芽巴贝斯虫内转录间隔区（BbiITS）、环形泰勒虫内转录间隔区（TaITS）、ToMPSP和AmMSP4基因的片段进行测序以进行系统发育分析。PCR结果显示，边缘无形体、环形泰勒虫、双芽巴贝斯虫和东方泰勒虫的总体感染率分别为29.1%、18.9%、11.2%和5.6%。在196份牛样本中有29份（15.1%）检测到两种或三种病原体的共同感染。序列分析结果表明，BbiRAP - 1a、BbiITS、Tams - 1、ToMPSP和AmMSP4在土耳其样本中具有保守性，序列同一性值分别为99.76%、99 - 99.8%、99.34 - 99.78%、96.9 - 99.61%和99.42 - 99.71%。相比之下，土耳其TaITS基因序列相对多样，同一性值为92.3 - 96.63%。在系统发育分析中，来自土耳其的双芽巴贝斯虫分离株与来自其他国家的分离株位于同一进化枝。另一方面，基于环形泰勒虫ITS序列的系统发育分析显示，来自土耳其的环形泰勒虫分离株在基因型上存在显著差异。此外，来自土耳其样本的东方泰勒虫分离株被归类为MPSP 3型基因型。这是土耳其首次报道MPSP 3型。此外，来自土耳其的AmMSP4分离株与来自其他国家的分离株位于同一进化枝。本研究为了解蜱传疾病的流行病学提供了重要数据，有望改进土耳其蜱传疾病的诊断和控制方法。
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