湖南省不同种类蛇源裂头蚴线粒体pnad1基因序列多态性研究

doi:10.12140/j.issn.1000-7423.2019.04.014

中国寄生虫学与寄生虫病杂志 ›› 2019, Vol. 37 ›› Issue (4): 448-452.doi: 10.12140/j.issn.1000-7423.2019.04.014

湖南省不同种类蛇源裂头蚴线粒体pnad1基因序列多态性研究

谭磊¹(), 王爱兵¹, 孔小鲜¹, 梁轩², 贺峻琳¹, 李洁¹, 胡丹¹, 刘伟^1,^*()

1 湖南农业大学动物医学院,长沙410128
2 安化县农业和粮食局,益阳 413500

收稿日期:2018-12-20 出版日期:2019-08-30 发布日期:2019-09-05
通讯作者: 刘伟
作者简介:
作者简介：谭磊（1994-）,男,博士研究生,从事动物传染病诊断与防控研究。E-mail: 812442773@qq.com.
基金资助:
湖南省科技厅项目（No. 2016NK2104）;湖南省教育厅优秀青年基金项目（No. 17B126）;湖南省畜牧局项目资助（No. 2139999）

Polymorphism of mitochondrial pnad1 gene of Spirometra erinaceieuropaei sparganum isolated from different species of snake in Hunan Province

Lei TAN¹(), Ai-bing WANG¹, Xiao-xian KONG¹, Xuan LIANG², Jun-lin HE¹, Jie LI¹, Dan HU¹, wei LIU^1,^*()

1 College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China
2 Anhua Agricultural Bureau, Yiyang 413500, China

Received:2018-12-20 Online:2019-08-30 Published:2019-09-05
Contact: wei LIU
Supported by:
Supported by Hunan Science and Technology Department(No. 2016NK2104), Hunan Education Department Excellent Youth Fund (No. 17B126), Hunan Animal Husbandry Bureau(No. 2139999)

摘要/Abstract

摘要：

目的对湖南省不同种类蛇源裂头蚴进行形态学与分子生物学鉴定,并分析其线粒体烟酰胺腺嘌呤二核苷酸脱氢酶第1亚基基因部分序列（pnad1）多态性。方法对从湖南永州、郴州、岳阳等地区6种常见的野生蛇体内获得的11条裂头蚴（来自大王蛇3条、乌梢蛇2条、眼镜蛇1条、灰鼠蛇2条、滑鼠蛇2条和银环蛇1条）进行形态学鉴定,PCR扩增其线粒体pnad1,并测序,分析其核苷酸序列种内差异,并与已报道的迭宫属绦虫和其他属绦虫的相应序列进行同源性比较;以土耳其斯坦东毕吸虫为外群,用软件MEGA7.0采用邻接法构建系统进化树。结果从不同蛇类分离的11条虫体经形态学鉴定为裂头蚴。PCR扩增出11条裂头蚴分离株的nad1序列,片段大小为648 bp,种内遗传差异分别为0~1.4%。与GenBank中收录的欧猬迭宫绦虫分离株序列相似性为97.8%~99.3%,与其它绦虫的序列相似性均低于74.3%。系统进化树结果显示,本研究分离的不同蛇源裂头蚴分离株与已知欧猬迭宫绦虫聚为一支,与微小膜壳绦虫和阔节裂头绦虫亲缘关系较近,与其它绦虫分支相隔较远。结论湖南省不同蛇源裂头蚴分离株均为欧猬迭宫绦虫,但存在一定的遗传变异,且线粒体pnad1基因适合作为分子标记对蛇源裂头蚴进行遗传变异研究。

关键词: 蛇源裂头蚴, 线粒体, 线粒体烟酰胺腺嘌呤二核苷酸脱氢酶第1亚基基因部分序列基因, 系统发育研究

Abstract:

Objective Spirometra erinaceieuropaei sparganum larvae were isolated from different species of snake and identified by morphology and molecular biology. The polymorphism of partial mitochondrial NADH dehydrogenase subunits 1 gene(pnad1) amplified from these isolated spargana was analyzed for its potential as a species marker. Methods Eleven sparganum larvae of S. erinaceieuropaei were isolated from different species of wild snake [Elaphe carinata (3), Zaocys dhumnade (2), Naja (1), Ptyas korros (2), Ptyas mucosus (2) and Bungarus multicinctus(1)] in Yongzhou, Chenzhou, Yueyang of Hunan Province and identified by morphology and DNA sequence. The pnad1 gene was amplified from these spargana isolated from different snakes and sequenced. The obtained sequences were aligned to determine their nucleotide variations and compared with their homologues from other tapeworms, and then a phylogenetic tree was constructed accordingly using MEGA7.0. Results The 11 larvae were identified as Spirometra erinaceieuropaei spargana by their morphology under microscopy and DNA sequence. The pnad1 gene was amplified from these collected spargana with size of 648 bp. The sequence results showed that the amplified pnad1 sequences were almost identical with 0-1.4% variation, but with 97.8%-99.3% sequence identity with different isolations of Spirometra erinaceieuropaei deposited in GenBank, and with less than 74.3% identity with pnad1 sequence of other tapeworms. Constracted phylogenetic tree based on the pnad1 gene sequences showed that the Hunan isolates were clustered in the same clade of Spirometra erinaceieuropaei, genetically closer to Dibothriocephalus latus and Hymenolepis nana, but further to other tapeworms. Conclusion The sparganum larvae isolated from different species of snakes in Hunan in the present study were identified as Spirometra erinaceieuropaei, and the mitochondrial pnad1 gene can be used as a genetic marker to identify tapeworm species.

Key words: Snake sparganum, Mitochondrial, Partial mitochondrial NADH dehydrogenase subunits 1 gene, Phylogenetic analysis

中图分类号:

R383.31

谭磊, 王爱兵, 孔小鲜, 梁轩, 贺峻琳, 李洁, 胡丹, 刘伟. 湖南省不同种类蛇源裂头蚴线粒体pnad1基因序列多态性研究[J]. 中国寄生虫学与寄生虫病杂志, 2019, 37(4): 448-452.

Lei TAN, Ai-bing WANG, Xiao-xian KONG, Xuan LIANG, Jun-lin HE, Jie LI, Dan HU, wei LIU. Polymorphism of mitochondrial pnad1 gene of Spirometra erinaceieuropaei sparganum isolated from different species of snake in Hunan Province[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2019, 37(4): 448-452.

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参考文献 19

[1]	谭磊, 王爱兵, 周湘, 等. 基于ITS基因分析黑斑蛙裂头蚴种系发育关系[J]. 中国兽医学报, 2017, 37(10): 1924-1927.
[2]	卢艳, 陈家旭, 李浩, 等. 上海市动物园蛇曼氏裂头蚴感染情况[J]. 中国寄生虫学与寄生虫病杂志, 2018, 36(6):593-596.
[3]	Hong Q, Feng JP, Liu HJ, et al. Prevalence of Spirometra mansoni in dogs, cats, and frogs and its medical relevance in Guangzhou, China[J] Int J Infect Dis, 2016, 53: 41-45.
[4]	周庆安, 全琛宇, 曾芸, 等. 广西南宁市市售蛙和蛇感染裂头蚴情况的调查[J]. 动物医学进展, 2013(11): 126-128.
[5]	曾利娟, 刘国华, 廖春玉, 等. 猬迭宫绦虫裂头蚴线粒体1基因部分序列的多态性研究[J]. 中国畜牧兽医, 2011, 38(3): 92-94.
[6]	曾庆仁, 贺美, 王芳, 等. 10例疑似曼氏裂头蚴病的病原学诊断[J]. 中国寄生虫学与寄生虫病杂志, 2012, 30(3): 224-232.
[7]	沈美清, 刘建, 杨利群, 等. 人感染曼氏裂头蚴病4例诊治分析[J]. 中国寄生虫学与寄生虫病杂志, 2015, 33(2): 125-126.
[8]	张玲玲, 吴斌, 丰燕, 等. 人体感染曼氏迭宫绦虫成虫的病原体鉴定和流行病学分析[J]. 中国寄生虫学与寄生虫病杂志, 2018, 36(2): 199-200.
[9]	李雯雯, 李健, 李树清, 等. 桂林蛇源裂头蚴分离株的分子鉴定及种系发育关系分析[J]. 动物医学进展, 2011, 32(10): 28-32.
[10]	李金福, 刘艳丹, 陈艳, 等. 贵州省安顺地区蛇源裂头蚴分离株的分子鉴定及种系发育关系分析[J]. 中国人兽共患病学报, 2016, 32(4): 376-382.
[11]	Dai RS, Liu GH, Song HQ, et al. Sequence variability in two mitochondrial DNA regions and internal transcribed spacer among three cestodes infecting animals and humans from China[J]. J Helminthol, 2012, 86(2): 7.
[12]	Liu W, Zhao GH, Tan MY, et al. Survey of Spirometra erinaceieuropaei spargana infection in the frog Rana nigromaculata of the Hunan Province of China[J]. Vet Parasitol, 2010, 173(1-2): 152-156.
[13]	Wei T, Zhang X, Cui J, et al. Levels of sparganum infections and phylogenetic analysis of the tapeworm Spirometra erinaceieuropaei sparganum in wild frogs from Henan Province in central China[J]. J Helminthol, 2015, 89(4): 433-438.
[14]	Zhang X, Wang H, Cui J, et al. Characterisation of the relationship between Spirometra erinaceieuropaei and Diphyllobothrium species using complete cytb and cox1 genes[J]. Infect Genet Evol, 2015, 35: 1-8.
[15]	Liu GH, Li C, Li JY, et al. Characterization of the complete mitochondrial genome sequence of Spirometra erinaceieuropaei (Cestoda : Diphyllobothriidae) from China[J]. Int J Biol Sci, 2012, 8(5): 640-649.
[16]	Zhang X, Duan JY, Shi YL, et al. Comparative mitochondrial genomics among Spirometra(Cestoda : Diphyllobothriidae) and the molecular phylogeny of related tapeworms[J]. Mol Phylogenet Evol, 2017, 117: 75-82.
[17]	马睿麟, 娄忠子, 李立, 等. 青海省细粒棘球蚴线粒体cox1基因和nad1基因多态性的研究.[J]. 中国兽医科学, 2014(12): 1251-1256.
[18]	杨俊萍, 郭书林, 陈信忠, 等. 多房棘球蚴线粒体NADH脱氢酶1基因分析及PCR检测方法[J]. 国际医学寄生虫病杂志, 2012, 39(3): 155-158.
[19]	郝桂英. 猪囊尾蚴西昌分离株线粒体cox1和nad1基因的序列测定与种系发育分析[J]. 中国畜牧兽医, 2015, 42(1): 61-66.

湖南省不同种类蛇源裂头蚴线粒体pnad1基因序列多态性研究

Polymorphism of mitochondrial pnad1 gene of Spirometra erinaceieuropaei sparganum isolated from different species of snake in Hunan Province

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