骆驼源片形吸虫的鉴定及遗传多样性研究  

Identification and genetic diversity analysis of camel-derived Fasciola sp.

作　　者：邱启官 陈淞楠 孟沁 谭笑若 李宁倩 刘晓恒 陈叔玉 刘伟[2] QIU Qi-guan;CHEN Song-nan;MENG Qin;TAN Xiao-ruo;LI Ning-qian;LIU Xiao-heng;CHEN Shu-yu;LIU Wei(Changsha Ecological Zoo,Changsha 410029,China;College of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]长沙生态动物园,湖南长沙410029 [2]湖南农业大学动物医学院,湖南长沙410128

出　　处：《中国预防兽医学报》2024年第12期1298-1302,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：湖南省自然科学基金(2021JJ30335)。

摘　　要：为了鉴定骆驼源片形吸虫的种类,研究其遗传多样性,本实验对6条骆驼源片形吸虫分离株的线粒体pcox1和pnad4,以及核糖体ITS基因进行PCR扩增、测序以及遗传多样性分析,并利用pcox1、pnad4和ITS基因构建系统发育树分析其遗传进化关系。结果显示,6条骆驼源片形吸虫pcox1、pnad4和ITS基因分别长342 bp、830 bp和748 bp,碱基差异率分别为0~0.6%、0~0.5%和0.1%~1.1%,种内高度保守;分别检测到2、4和5个单倍型,核苷酸多样性分别为0.003±0.003、0.002±0.003和0.005±0.005;与其他国家或地区的大片吸虫相应基因序列的同源性分别为99.7%~100.0%、95.9%~100.0%和98.9%~100.0%,与肝片吸虫相应基因序列的同源性分别为92.7%~93.3%、86.7%~87.3%和98.1%~99.2%,与其他片形吸虫相应基因序列的同源性分别为89.5%~89.8%、80.0%~80.2%和94.5%~95.6%。3个基因的系统进化分析结果显示,6条骆驼源片形吸虫均与已知大片吸虫分离株聚类形成同一分支,与肝片吸虫、杰氏片形吸虫处于不同分支。上述研究结果表明6条骆驼源片形吸虫分离株为大片吸虫,pcox1、pnad4和ITS基因均可作为大片吸虫的遗传分子标记。本研究为大片吸虫的分类鉴定以及骆驼源片形吸虫感染的防治提供参考依据。In order to identify the species and study the genetic diversity of camel-derived Fasciola,mitochondrial pcox1 and pnad4 and ribosomal ITS of six camel-derived Fasciola isolates were amplified by PCR,sequenced and genetic diversity analyzed.A phylogenetic tree was constructed using pcox1,pnad4 and ITS genes to analyze their genetic evolutionary relationships.The results revealed that the lengths of pcox1,pnad4,and ITS genes in the camel-derived Fasciola isolates were 342bp,830bp,and 748bp,respectively,with nucleotide diversity rates ranging from 0 to 0.6%,0 to 0.5%,and 0.1%to 1.1%,respectively,indicating highly conserved within the species.Haplotype analysis identified two,four,and five haplotypes in these three genes,with nucleotide diversities of 0.003±0.003,0.002±0.003,and 0.005±0.005,respectively.The genetic similarity between the camel-derived Fasciola isolates and those from other countries/regions was 99.7%to 100.0%,95.9%to 100.0%,and 98.9%to 100.0%,respectively.Comparison with Fasciola hepatica demonstrated homogeneity levels of 92.7%to 93.3%,86.7%to 87.3%,and 98.1%to 99.2%,respectively,while similarity with other Fasciola species ranged from 89.5%to 89.8%,80.0%to 80.2%,and 94.5%to 95.6%,respectively.Phylogenetic analysis indicated that the six camel-derived Fasciola isolates clustered into the same branch as the known Fasciola hepatica isolates,and were in different branches from Fasciola gigantica and Fasciola jacksoni.These results showed that the six camel-derived Fasciola isolates were Fasciola hepatica,and pcox1,pnad4 and ITS genes could be used as genetic molecular markers.This study provides a theoretical basis for the identification,prevention and control of camel fascioliaisi.

关 键 词：片形吸虫 骆驼 pcox1 pnad4 ITS 分子鉴定 遗传多样性 

分 类 号：S852.6[农业科学—基础兽医学]