ABSTRACT

Theileriosis, an important tick-borne hemoprotozoan disease that can reduce production and productivity, results in severe economic loss in goat industry. Due to lack of facilities and interest of the researchers the sub-clinical form of theileriosis caused by Theileria luwenshuni was under diagnosed in Assam, India. Therefore, a study was carried out to determine the prevalence of theileriosis in goats from Kamrup (Metro) district of Assam. A total of 454 blood samples collected from goats revealed 39.87 % (181/454) infection with Theileria sp. The genomic DNA extracted from blood samples amplified for a fragment of 18S ribosomal RNA (18S rRNA) gene employing polymerase chain reaction (PCR) using Theileria genus specific primers showed distinct bands at 1098 bp in agarose gel. The resultant PCR products were sequenced and BLAST analysis indicated 99.9% homology with T. luwenshuni. Parasitological methods and PCR technique followed by sequence analysis suggest infection with T. luwenshuni was associated with theileriosis in investigated goats. The results of the present study showed PCR to be more efficient in detection of Theileria positive cases than the microscopic method.

Abbreviations: rRNA: Ribosomal ribonucleic acid; PCR: polymerase chain reaction; ml: milliliter; EDTA: Ethylene Diammine Tetraacetic Acid; °C: degree Celsius; DNA: Deoxyribonucleic acid; ME: microscopic examination; sp.: species; V-volt; µg: micro gram; TAE: Tris-acetate-EDTA; F: forward; R: reverse; NCBI: National Center for Biotechnology Information; BLAST-n Basic Local Alignment Search Tool for nucleotide.

摘要

泰勒虫病是一种重要的蜱传血原生动物疾病，会降低产量和生产力，导致山羊产业严重经济损失。由于缺乏设施和研究人员的兴趣，由Theileria luwenshuni引起的亚临床形式的泰勒虫病在印度阿萨姆邦被诊断出来。因此，进行了一项研究，以确定阿萨姆邦卡姆鲁普（Metro）地区山羊的泰勒病患病率。从山羊身上采集的总共 454 份血液样本显示 39.87 % (181/454) 感染了泰勒菌。从血液样本中提取的基因组 DNA 通过使用泰勒菌的聚合酶链反应 (PCR) 扩增 18S 核糖体 RNA (18S rRNA)基因片段属特异性引物在琼脂糖凝胶中在 1098 bp 处显示出明显的条带。对所得PCR产物进行测序，BLAST分析表明与T. luwenshuni有99.9%的同源性。寄生虫学方法和 PCR 技术以及随后的序列分析表明T. luwenshuni感染与研究山羊的泰勒虫病有关。本研究结果表明，PCR 在检测泰勒菌阳性病例方面比显微镜方法更有效。

缩写： rRNA：核糖体核糖核酸；PCR：聚合酶链式反应；ml：毫升；EDTA：乙二胺四乙酸；°C：摄氏度；DNA：脱氧核糖核酸；ME：显微镜检查；sp.：物种；V-伏；µg：微克；TAE：Tris-醋酸盐-EDTA；F：前锋；R：反向；NCBI：国家生物技术信息中心；BLAST-n 基本的核苷酸局部比对搜索工具。