保定地区奶牛源隐孢子虫的分离与基因鉴定

发布时间：2018-03-05 10:34

本文选题：奶牛　切入点：牛隐孢子虫　出处：《黑龙江畜牧兽医》2017年15期 　论文类型：期刊论文

【摘要】：为了解保定地区奶牛源隐孢子虫感染情况,从当地3个奶牛场随机采集145份奶牛粪便经饱和蔗糖溶液漂浮后直接镜检和抗酸染色后镜检,阳性样品采用PCR方法扩增18S rRNA基因,同时将扩增出的目的片段进行克隆和序列分析,并将分离株与其他11个隐孢子虫参考株的18S rRNA序列进行比对和遗传距离比较。结果表明:有10份粪便样本为隐孢子虫阳性,感染率为6.9%(10/145),不同奶牛场、年龄段奶牛隐孢子虫感染率有显著性差异(P0.05)。10份粪便样品采用PCR方法扩增后有7份为18S rRNA基因阳性,扩增出的18S rRNA部分基因片段长528 bp。保定地区隐孢子虫分离株扩增的基因序列与牛源隐孢子虫18S rRNA基因序列(Gen Bank登录号为HQ179571)同源性最高,确定保定地区分离到的奶牛源隐孢子虫为牛隐孢子虫。
[Abstract]:In order to understand the infection of Cryptosporidium from dairy cattle in Baoding area, 145 samples of cow feces were collected randomly from three dairy farms in Baoding. The 18s rRNA gene was amplified by PCR method after floating in saturated sucrose solution and acid fast staining. The amplified fragments were cloned and sequenced. The 18s rRNA sequences of the isolates were compared with those of the other 11 Cryptosporidium reference strains. The results showed that 10 fecal samples were Cryptosporidium positive. The infection rate of Cryptosporidium in different dairy farms was significantly different from that of 10 / 145g in dairy cattle. Seven of the 10 fecal samples amplified by PCR were 18s rRNA positive, and the infection rate of Cryptosporidium was significantly higher than that of the control, and there was significant difference in the infection rate of Cryptosporidium in dairy cattle. The amplified partial gene fragment of 18s rRNA was 528bp.The amplified gene sequence of Cryptosporidium isolated from Baoding area was the highest homology with the accession number HQ179571 of the 18s rRNA gene sequence of Cryptosporidium bovis (HQ179571). Cryptosporidium was identified as Cryptosporidium bovis in Baoding area.
【作者单位】：河北农业大学动物医学院;
【基金】：河北省现代农业产业技术体系“奶牛产业创新团队建设项目奶牛疾病防控技术研发与示范”(HBCT2013080204)
【分类号】：S858.23

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