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首页> 外文期刊>Journal of Parasitology >First Isolation of Besnoitia besnoiti From a Chronically Infected Cow in Spain
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First Isolation of Besnoitia besnoiti From a Chronically Infected Cow in Spain

机译：首次从西班牙的慢性感染牛中分离出甜菜疯

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Besnoitia besnoiti was isolated from a skin biopsy of a chronically infected cow from central Spain. Zoites released from macroscopic cysts were adapted to its culture in vitro on a MARC-145 cell monolayer. Tachyzoites produced in vitro were either cryopreserved or used for genomic DNA isolation. A 2206 nt sequence containing 18S ribosomal RNA gene, internal transcribed spacer 1 (ITS1), and a partial sequence of 5.8S ribosomal RNA gene was amplified by PCR and sequenced. This sequence showed a 99-100% identity to 18S, ITS1, and 5.8S sequences of B. besnoiti published in databases. After analysis by transmission and scanning electron microscopy of isolated bradyzoites and tachyzoites, it was observed that their ultrastructural morphology coincided with B. besnoiti. The isolate characterized in this study was identified as B. besnoiti on the basis of the disease produced, molecular characteristics, and morphology. The B. besnoiti isolate was denoted its BbSpain-1; it is the first isolate obtained and characterized in Spain and one of the first European isolates adapted to grow in vitro. The isolation and in vitro production of this B. besnoiti isolate offers a good opportunity to study general aspects of bovine besnoitiosis. including epidemiology, pathogenesis, and diagnosis of this re-emergent disease.
机译：贝诺氏菌（Besnoitia besnoiti）是从西班牙中部一名慢性感染母牛的皮肤活检中分离出来的。从肉眼可见的囊肿中释放出的沸石可在MARC-145细胞单层上进行体外培养。将体外产生的速殖子冷冻保存或用于基因组DNA分离。通过PCR扩增了包含18S核糖体RNA基因，内部转录间隔区1（ITS1）和5.8S核糖体RNA基因的部分序列的2206nt序列。该序列与数据库中公布的贝氏芽孢杆菌的18S，ITS1和5.8S序列具有99-100％的同一性。通过透射和扫描电子显微镜分析分离的缓殖子和速殖子后，观察到它们的超微结构形态与贝氏芽孢杆菌一致。根据产生的疾病，分子特征和形态，本研究中鉴定出的分离株被鉴定为B. besnoiti。贝氏芽孢杆菌分离株被命名为BbSpain-1。它是在西班牙获得并鉴定的第一个分离株，也是最早适合在体外生长的欧洲分离株之一。该B. besnoiti分离物的分离和体外生产为研究牛Bennoitiosis的一般方面提供了一个很好的机会。包括这种新发疾病的流行病学，发病机制和诊断。

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