奶牛源微小隐孢子虫的分子鉴定及动物感染试验  

Identification of Cryptosporidiumspecies in dairy cattle and oocyst collection by animal experiment

作　　者：齐萌[1] 黄建营 王臣荣 余复昌 李俊强[1] 张龙现[1] 

机构地区：[1]河南农业大学牧医工程学院/河南省人兽共患病国际联合实验室,河南郑州450002

出　　处：《中国兽医学报》2016年第1期85-89,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金重大国际合作项目(31110103901);河南省高校科研创新团队项目(012IRTSTHN005)

摘　　要：采用饱和蔗糖溶液漂浮法检查商丘市某奶牛场牛新鲜粪便样本的隐孢子虫卵囊,用18SrRNA基因对隐孢子虫进行PCR扩增和限制性片段长度多态性分析;基于GP60基因位点对微小隐孢子虫进行基因亚型鉴定。结果显示:103份样本中有50份为隐孢子虫阳性,42份经形态学鉴定为安氏隐孢子虫,8份形态学未能鉴定到种。经限制性片段长度多态性分析,7个分离株为微小隐孢子虫,1个分离株为牛隐孢子虫;序列比对分析,7个微小隐孢子虫均为人兽共患基因亚型IIdA19G1。接种1头3日龄犊牛1×106个卵囊,潜隐期为3d,显露期为14d,于感染后第7天和第10天出现2个排卵囊高峰期,收集到大量纯卵囊。In order to identlly Cryptosporidium species m dairy cattle,and collect the oocysts of Cryptosporidium parvum. Fresh fecal samples collected from farm in Shangqiu were examinated by using the Sheather＇s sugar flotation technique, and the Cryptosporidium-positive samples were genotyped with a restriction fragment length polymorphism analysis of the small subunit riboso- mal RNA. Subtyping of C. parvum was identified using 60 000 glyeoprotein （gp60） gene. Among the 103 fecal samples, 50 were Crxptosporidiurn-positive. The 42 isolates were identified as C. andetsoni upon microscopic, and eight isolates were not identified the species of Cryptosporidi- urn. Th~ analysis of RLFP showed that seven isolates belonged to C. parvurn,whereas one isolate was C. bovis. Sequence analyses of gp60 gene revealed that seven isolates were zoontie subtype Ⅱ- dA19Gl One 3-day-old calf was inoculated by 1× 106 oocysts,with the latent period and the pa- tent peeiod were 3 days and 14 days,respectively. The two peaks of ooeyst shedding were noticed at 7 PID and 10 PID,respectively. A large number of C. parvurn oocysts were obtained.

关 键 词：微小隐孢子虫 鉴定 感染 奶牛 

分 类 号：S852.72[农业科学—基础兽医学]