秦岭大熊猫槽盘吸虫的鉴定与遗传进化分析

Identification and phylogenetic analysis of Ogmocotyle sp.from giant panda in Qinling Mountain

本研究旨在对采自大熊猫秦岭亚种(ailuropoda melanoleuca qinlingensis)小肠内的槽盘吸虫进行种类鉴定及遗传进化分析。通过形态学和分子生物学方法,对虫体样本分别进行染色显微镜观察和核糖体DNA内转录间隔区Ⅱ(ITS2)部分序列的PCR扩增和序列分析。经不同染色方法比较后,发现孔雀绿染色法处理后的虫体外部形态以及内部结构清晰,形态上符合印度槽盘吸虫(Ogmocotyle indica)的典型特征;序列及遗传进化分析表明,10个样本ITS2序列完全一致,其系统发生树显示10个样本与O.indica位于同一分枝。研究结果表明,分离的虫株从形态学和进化分析上均与印度槽盘吸虫具有高度一致性,提示该槽盘吸虫为印度槽盘吸虫。

The aim of the present study were to identify and genetically analyze Ogrnocotyle sp. col- lected from the small intestine of ailuropoda melanoleuca qinlingensis. Adult trematodes were firstly identified based on morphological observation under microscopy after different staining methods,and then were analyzed their second internal transcribed spacer （ITS2） by PCR and se- quencing. Comparison of different staining methods indicated that Malachite green worked clearly to show external and internal morphological features of parasites, and these characters were con- sistent with that of Ograocotyle indica. Sequencing and genetic analysis showed that ITS2 se-quences of ten samples were the same and they were located in the same clade with O. indica in phylogenetic tree based on ITS2 sequences. These results indicated that Ogrnocotyle sp. in the present study had high identity to O. indica, suggesting that Ogrnocotyle sp. collected from giant panda was O. indica.