Continuous in vitro cultivation of a recently identified Babesia that infects small ruminants in China
Continuous in vitro cultivation of a recently identified Babesia that infects small ruminants in China
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最近在中国发现的感染小型反刍动物的巴贝虫的连续体外培养
DOI：
10.1016/j.vetpar.2012.02.006
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发表时间：
2012-07-06
影响因子：
2.6
通讯作者：
Luo, Jianxun
中科院分区：
文献类型：
--
作者：
Guan, Guiquan;Ma, Miling;Luo, Jianxun
巴贝斯虫新疆分离自新疆的一只去脾绵羊，该绵羊由新疆的绵羊和牛群中采集的血缘管圆线虫和类囊线虫感染。根据18S核糖体RNA(18S RRNA)和内转录间隔区(ITS)基因序列的比对，认为它是一个新的绵羊巴贝斯虫种。用感染的绵羊血液建立绵羊寄生虫的体外连续培养。在含7.5%绵羊红细胞(RBC)的RPMI1640培养液中，在37℃和5%CO2的培养箱中，24孔板和6孔板中寄生虫红细胞(PPE)的百分率达到10%的峰值。在75 cm(2)的培养瓶中加入2.5%的RBC，相同的培养基数可增加到20-50%。巴贝斯虫两个无性系。新疆采用有限稀释法进行筛选。用微量分光光度法和PPE法测定了这些品系的体外生长特性。在绵羊红细胞内的世代时间为15.20~16.27h，并通过体外培养和体内感染鉴定了寄生虫的寄主范围。羊、牛、梅花鹿和人的红细胞在体外可被线虫侵入，但不能在人的红细胞内繁殖。这些寄生虫在体外不能进入山羊的红细胞。然而，在体内，只有绵羊才能被线虫感染。最后是一株巴贝斯虫。新疆类寄生虫(与巴贝斯虫原株同源性99.5%)。新疆)从甘肃省西部地区采集的19份绵羊血液样本中分离到1份，分离自中国。(C)2012爱思唯尔B.V.保留所有权利。
Babesia sp. Xinjiang was isolated from a splenectomised sheep infested by Rhipicephalus sanguineus and Hylomma anatolicum anatolicum, collected from sheep and cattle in Xinjiang province. It was considered to be a novel ovine Babesia species on the basis of its morphology, pathogenicity, vector tick species and alignments of 18S ribosomal RNA (18S rRNA) and internal transcribed spacers (ITS) gene sequences. Continuous in vitro cultures of the ovine parasite were established using infected sheep blood. In RPMI 1640 medium with 7.5% sheep red blood cells (RBCs) maintained in an incubator at 37 degrees C and 5% CO2, the percentage of parasitized erythrocytes (PPE) peaked at 10% in 24- and 6-well plates. It increased to 20-50% with the same culture medium but with 2.5% RBC in 75 cm(2) flasks. Two clonal lines of Babesia sp. Xinjiang were screened using the limiting dilution method. Growth characteristics of these lines in vitro were measured by a microtiter-based spectrophotometric method and from the PPE. The generation time in sheep erythrocytes was between 15.20 h and 16.27 h. Furthermore, the host range of parasite was identified with in vitro culture and in vivo infection. Erythrocytes of sheep, cattle, sika deer and humans could be invaded into by lines in vitro, but the parasites could not propagate in human erythrocytes. The parasites could not enter erythrocytes from goats in vitro. However, in vivo, only sheep could be infected by lines. Finally, a Babesia sp. Xinjiang-like parasite (which shared 99.5% identity with the original strain of Babesia sp. Xinjiang) was isolated using this in vitro culture system from 1 of 19 sheep blood samples collected from western Gansu province, China. (c) 2012 Elsevier B.V. All rights reserved.