Hao Pan, Yang Na, Cui Xia, Liu Jing, Yang Daoyu, Liu Qun
Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
J Parasitol. 2014 Dec;100(6):812-6. doi: 10.1645/14-498.1.
Neospora caninum is an important cause of abortion in cattle worldwide, but the isolation of a viable parasite from an abortus is difficult, and viable N. caninum has not been isolated from any host in China. In the present study, peripheral blood samples were collected from a jugular vein of an adult dairy cow that had aborted; the cow was seropositive to N. caninum antibodies by ELISA. White blood cells were separated and seeded onto Vero cell monolayer cultures for parasite isolation. Tachyzoites were first observed in cell culture on day 84 after initial inoculation. The parasite was confirmed to be N. caninum by gene sequencing and immunofluorescence, and by bioassays in BALB/c mice. The new N. caninum isolate (NC-Bj) has a unique pattern on microsatellite Cont-14. To our knowledge, this is the first successful isolation of N. caninum in China from any host.
犬新孢子虫是全球范围内牛流产的一个重要病因，但从流产胎儿中分离出活的寄生虫很困难，在中国尚未从任何宿主中分离出活的犬新孢子虫。在本研究中，从一头流产的成年奶牛颈静脉采集外周血样本；通过酶联免疫吸附测定（ELISA），该奶牛对犬新孢子虫抗体呈血清阳性。分离白细胞并接种到Vero细胞单层培养物上以进行寄生虫分离。在初次接种后第84天，在细胞培养物中首次观察到速殖子。通过基因测序、免疫荧光以及在BALB/c小鼠中的生物测定，确认该寄生虫为犬新孢子虫。新的犬新孢子虫分离株（NC-Bj）在微卫星Cont-14上具有独特的图谱。据我们所知，这是中国首次成功从任何宿主中分离出犬新孢子虫。
