前后盘吸虫梅花鹿源分离株的种类鉴定及遗传进化分析

Identification and Phylogenetic Analysis of Paramphistomum cervi fromCervus nippon

旨在研究温州地区的梅花鹿源前后盘吸虫的种类和遗传进化,利用形态学和分子生物学方法,对虫体样本进行染色显微镜观察及pcox1和pnad1序列的PCR扩增和序列分析。形态学鉴定结果表明,前后盘吸虫梅花鹿源分离株在外部形态、大小以及内部组织器官与鹿前后盘吸虫(Paramphistomum cervi)相近。序列比较分析结果表明,3株前后盘吸虫梅花鹿源分离株样本的pcox1和pnad1序列完全一致,扩增产物长度分别为446和505bp,与GenBank中鹿前后盘吸虫(KT198987)同源性最高,分别为98.2%和97.7%。遗传进化分析表明,3株前后盘吸虫梅花鹿源分离株与鹿前后盘吸虫位于同一分支。表明,分离的虫株在形态学和遗传进化分析上均与鹿前后盘吸虫具有高度的一致性,提示获得的前后盘吸虫梅花鹿源分离株为鹿前后盘吸虫。

The aim of the present study was to identify and genetically analyze Paramphistomum sp.collected from Cervus nippon around Wenzhou region of Zhejiang province in China.Adult worms were identified based on morphological observation under microscopy after staining,and then the pcox1 and pnad1 genes were analyzed by PCR and sequencing.Morphological identification results showed that Paramphistomum sp.from C.nippon were close to P.cervi in external shape,size and internal organs.Comparative sequence analysis demonstrated that thepcox1 andpnad1 sequences from three C.nippon-derived Paramphistomum sp.were 446 bp and 505 bp in length and entirely consistent in similarity.Moreover,the homologies were the highest between isolates from C.nippon and P.cervi （KT198987） in GenBank,reached 98.2% and 97.7%,respectively.Phylogenetic analysis revealed that the three Paramphistomum samples from C.nippon and the P.cervi （KT198987） locate in the same branch.These results indicated that the Paramphistomum sp.has high consistency with P.cervi in morphology and phylogenetic analysis,suggesting the Paramphistomum sp.sampled from the C.nippon in Wenzhou area is P.cervi.