山羊腔阔盘吸虫的形态学观察和分子鉴定

Morphological Observation and Molecular Identification of Eurytrema coelomaticum in Goats

为了从形态学和分子水平对山羊腔阔盘吸虫(Eurytrema coelomaticum)的种类鉴定提供依据,对采集自广东佛山发病山羊胰脏的阔盘吸虫在形态学观察的基础上,抽提虫体基因组DNA,通过PCR方法对虫体核糖体18SrRNA基因和线粒体cox1基因进行扩增,将产物与pMD18-T载体连接,转化到大肠埃希菌DH5α感受态细胞进行克隆、测序和序列分析。结果获得虫体18SrRNA基因的大小为1 922bp,线粒体cox 1基因大小为444bp。序列分析显示,获得的18SrRNA基因与GenBank收录的腔阔盘吸虫(E.coelomaticum,登录号为DQ401035)同源性高达99%;线粒体cox1基因与GenBank收录的胰阔盘吸虫(E.pancreaticum,登录号为KC535544)的同源性仅为90%。研究结果从分子水平证明采集自广东佛山山羊体的阔盘吸虫为腔阔盘吸虫(E.coelomaticum),对山羊阔盘吸虫病的防控有指导意义。

In order to provide the basis for the identification of Eurytrema coelomaticum in goats from morphological and molecular level,genomic DNA of the parasites,which were collected from the pancreas of a goat with clinical symptoms in Foshan,Guangdong province,was extracted based on the morphological observation.Then PCR was used to amplify the 18S rRNA gene and cox1 gene of this parasite.The amplified products were then ligated with PMD18-T vector and transformed into Escherichia coli DH5ɑ for cloning and sequencing.The 18S rRNA gene sequence was 1 922 bp,and the cox 1 gene sequence was 444 bp.Based on BLAST analysis,the 18S rRNA gene sequence had 99% similarity to E.coelomaticum in GenBank (accession no DQ401035),and the cox 1 gene sequence had 90% similarity to E.pancreaticum (accession no KC535544).The results proved that the parasites collected from the pancreas of a goat were molecularly identified as E.coelomaticum ,which is of significance for the prevention and control of E.coelomaticum in goats.