Isolation of Babesia divergens from carrier cattle blood using in vitro culture
Isolation of Babesia divergens from carrier cattle blood using in vitro culture
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DOI：
10.1051/vetres:2003047
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发表时间：
2004-01-01
影响因子：
4.4
通讯作者：
Chauvin, A
中科院分区：
文献类型：
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作者：
Malandrin, L;L'Hostis, M;Chauvin, A
分歧巴贝斯虫是西欧牛巴贝斯虫病的主要病原体，从自然感染的牛中分离得到。通过绵羊红细胞体外培养技术检查了96份血样：其中19份采集自疾病急性期的动物，血涂片上可见寄生虫血症，而其余77只动物未显示显微镜可检测到的寄生虫。B。从19个最初的血液样品以及从无症状动物收集的31个样品中培养趋异菌。在培养物中检测到寄生虫之前的时间段在后一种样品中从6天到20天不等。对采样条件（抗凝剂）和保存时间的影响进行了试验。免疫荧光抗体检测和培养之间获得了良好的相关性，从无症状动物中采集的样品的相同结果（阳性或阴性）为89.6%。测定了体外培养方法的灵敏度，使用三种不同分离株进行的三项独立实验的灵敏度约为10个寄生虫/mL全血，证实了其适用于检测和培养多种B。分离自携带者牛的分歧菌。在自然感染的动物的血液中，在急性巴贝西虫病阶段后9个月确实可以分离出寄生虫。本研究中收集的50株分离株使用相同的培养基成功传代、冻存和复苏。体外分离B。从无症状携带者牛中分离寄生虫，并将允许分析牛群中的寄生虫多样性。
Babesia divergens, the main causative agent of bovine babesiosis in Western Europe, was isolated from naturally infected cattle. Ninety-six blood samples were examined by means of an in vitro culture technique in sheep erythrocytes: 19 of them were collected from animals in the acute phase of the disease with visible parasitemia on blood smears, while the 77 remaining animals showed no microscopically detectable parasites. B. divergens was cultured from the 19 first blood samples as well as from 31 samples collected from asymptomatic animals. The time period before parasites could be detected in the culture varied in the latter samples from 6 to 20 days. The effects of sampling condition (anticoagulant used) and storage length were tested. A good correlation was obtained between immunofluorescent antibody test and culture, with identical results (positive or negative) for 89.6% of the samples collected from asymptomatic animals. The sensitivity of the in vitro culture method was determined and was about 10 parasites/mL of whole blood from three independent experiments performed with three different isolates, confirming its suitability to detect and culture diverse B. divergens isolates from carrier cattle. The parasites could indeed be isolated 9 months after the acute babesiosis phase in the blood of naturally infected animals. The 50 isolates collected in this study were successfully subcultured, cryopreserved and resuscitated using the same culture medium. The in vitro isolation of B. divergens from asymptomatic carrier cattle was achieved and will allow the analysis of parasite diversity within cattle herds.