上海市动物园蛇曼氏裂头蚴感染情况
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摘要
目的了解上海市动物园蛇曼氏裂头蚴感染情况。方法 2017年对上海市动物园送检的30条蛇进行解剖检查,分离曼氏裂头蚴,记录寄生部位和数量;新鲜虫体经盐酸卡红染色后,于显微镜下进行形态学鉴定。提取虫体基因组DNA, PCR扩增曼氏裂头蚴线粒体基因细胞色素c氧化酶亚单位1 (cytochrome c oxidase subunit 1, cox1)、细胞色素b (cytochrome b, cytb),并进行测序分析。结果上海市动物园送检蛇的裂头蚴总感染率为86.7%(26/30),共检获裂头蚴626条,感染度为24.1条/蛇。其中6条王锦蛇中有3条感染裂头蚴,感染度为1.33条/蛇;乌梢蛇的感染率为95.8%(23/24),感染度为27.0条/蛇。裂头蚴主要寄生于蛇肌肉组织,占总数的52.9%(331/626)。显微镜下观察,所获虫体具有曼氏裂头蚴的形态特征。cox1基因和cytb基因的PCR扩增产物分别约为470 bp和1 400 bp。对26条蛇中分离的裂头蚴cox1和cytb基因序列一致性分析结果显示,cox1基因的序列一致性为96.4%～100%, cytb基因的为97.8%～100%。与GenBank中的绦虫序列进行比对,发现其与欧猥迭宫绦虫(GenBank登录号为KP738288、 AB374543)的序列一致性为99.8%～100%,与其他种类绦虫的序列一致性为54.2%～84.4%。结论上海市动物园送检蛇的曼氏裂头蚴感染率较高,蛇的曼氏裂头蚴可感染人类,需加强防控。
Objective To investigate the infection status of plerocercoid in snakes from Shanghai Zoo in order to provide information for the prevention and treatment of sparganosis. Methods Thirty snakes were provided from Shanghai Zoo and dissected in 2017. The plerocercoids were collected from the snakes, the number of plerocercoids and their location were recorded. The structure of plerocercoid was examined under microscope after being Carmine stained. The total DNA was extracted from the plerocercoids and the DNA fragments for cytochrome c oxidase subunit 1(cox1) and cytochrome b(cytb) genes were amplified by PCR. The amplified PCR products were sequenced and analyzed. Results The infection rate of plerocercoid in the snakes was 86.7%(26/30) with total626 plerocercoids found, with an average intensity of infection of 24.1 plerocercoids per snake. The infection rate in snake Elaphe carinata and Zaocys dhumnades was 50.0%(3/6) and 95.8%(23/24), with the average intensity of infection of 1.3 and 27.0 plerocercoids per snake, respectively. Muscular tissue was the most common parasitizing location for the plerocercoids, accounting for 52.9%(331/626). The isolated worms were in accordance with Spirometra mansoni plerocercoids in morphology. The PCR amplified product of cox1 gene was about 470 bp and cytb gene was about 1 400 bp, respectively, sharing 96.4% ～100%(cox1) and 97.8%～100%(cytb) sequence identity among the 26 samples. In comparison with the different tapeworm sequences in GenBank, it was found that the amplified cox1 and cytb gene sequences in the plerocercoids were highly homologous to those of S. erinaceieuropaei(99.8% ～100%, GenBank Accession No. KP738288, AB374543) and less homologous to other species of tapeworm.Conclusion The snakes collected from Shanghai Zoo are widely infected with S. mansoni plerocercoids, indicating that they can be the infection source of human sparganosis.
Objective To investigate the infection status of plerocercoid in snakes from Shanghai Zoo in order to provide information for the prevention and treatment of sparganosis. Methods Thirty snakes were provided from Shanghai Zoo and dissected in 2017. The plerocercoids were collected from the snakes, the number of plerocercoids and their location were recorded. The structure of plerocercoid was examined under microscope after being Carmine stained. The total DNA was extracted from the plerocercoids and the DNA fragments for cytochrome c oxidase subunit 1(cox1) and cytochrome b(cytb) genes were amplified by PCR. The amplified PCR products were sequenced and analyzed. Results The infection rate of plerocercoid in the snakes was 86.7%(26/30) with total626 plerocercoids found, with an average intensity of infection of 24.1 plerocercoids per snake. The infection rate in snake Elaphe carinata and Zaocys dhumnades was 50.0%(3/6) and 95.8%(23/24), with the average intensity of infection of 1.3 and 27.0 plerocercoids per snake, respectively. Muscular tissue was the most common parasitizing location for the plerocercoids, accounting for 52.9%(331/626). The isolated worms were in accordance with Spirometra mansoni plerocercoids in morphology. The PCR amplified product of cox1 gene was about 470 bp and cytb gene was about 1 400 bp, respectively, sharing 96.4% ～100%(cox1) and 97.8%～100%(cytb) sequence identity among the 26 samples. In comparison with the different tapeworm sequences in GenBank, it was found that the amplified cox1 and cytb gene sequences in the plerocercoids were highly homologous to those of S. erinaceieuropaei(99.8% ～100%, GenBank Accession No. KP738288, AB374543) and less homologous to other species of tapeworm.Conclusion The snakes collected from Shanghai Zoo are widely infected with S. mansoni plerocercoids, indicating that they can be the infection source of human sparganosis.
引文
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[13]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,2013,34(11):126-128.
[14]蒋红涛,陈艳,吴泽江,等.贵州省部分地区蛙、蛇体内曼氏裂头蚴感染情况的调查[J].贵州师范大学学报(自然科学版),2008,26(1):5-6,65.
[15]Hu M,Gasser RB.Mitochondrial genomes of parasitic nematodes-progress and perspectives[J].Trends Parasitol,2006,22(2):78-84.
[16]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.
[17]郝桂英.线粒体基因在带属绦虫分子分类中的研究进展[J].动物医学进展,2014,35(1):94-98.
[18]郝桂英,杨应东,周英姿,等.基于线粒体cox1和cytb基因对四川地区多头带绦虫的种群遗传多样性研究[J].畜牧兽医学报,2014,45(4):631-638.
[19]Zhang X,Cui J,Liu LN,et al.Genetic structure analysis of Spirometra erinaceieuropaei isolates from central and southern China[J].PLoS One,2015,10(3):e0119295.
[2]裘明华,裘明德.人裂头蚴病和无头蚴病:Ⅱ.病理学、临床、流行病学及控制的过去和现在[J].中国寄生虫学与寄生虫病杂志,2009,27(3):251-260.
[3]吴观陵.人体寄生虫学[M].4版.北京:人民卫生出版社2013:516-531.
[4]蔺西萌,王中全.我国曼氏裂头蚴病临床特征概述[J].中国病原生物学杂志,2011,6(6):467-468,471.
[5]Wang F,Zhou L,Gong S,et al.Severe infection of wild-caught snakes with Spirometra erinaceieuropaei from food markets in Guangzhou,China involves a risk for zoonotic sparganosis[J]J Parasitol,2011,97(1):170-171.
[6]Li MW,Song HQ,Li C,et al.Sparganosis in mainland China[J].Int J Infect Dis,2011,15(3):e154-156.
[7]钟睿翀,沈浩贤,张莉.形态学实用技术[M],北京:中国医药科技出版社,2007:72-73.
[8]Boonyasiri A,Cheunsuchon P,Srirabheebhat P,et al.Sparganosis presenting as cauda equina syndrome with molecular identifica-tion of the parasite in tissue sections[J].Korean J Parasitol,2013,51(6):739-742.
[9]Zhang X,Wang H,Cui J,et al.The phylogenetic diversity of Spirometra erinaceieuropaei isolates from southwest China revealed by multi genes[J].Acta Trop,2016,156:108-114.
[10]Lu G,Shi DZ,Lu YJ,et al.Retrospective epidemiological analysis of sparganosis in mainland China from 1959 to 2012[J].Epidemiol Infect,2014,142(12):2654-2661.
[11]吴有陵,朱顺海,董辉,等.上海市大王蛇、赤练蛇感染寄生虫情况初步调查[J].上海师范大学学报(自然科学版),2013,42(6):629-634.
[12]杨光大,王付民,龚世平,等.广东省蛇类曼氏裂头蚴感染分析[J].广东林业科技,2015,31(5):80-83.
[13]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,2013,34(11):126-128.
[14]蒋红涛,陈艳,吴泽江,等.贵州省部分地区蛙、蛇体内曼氏裂头蚴感染情况的调查[J].贵州师范大学学报(自然科学版),2008,26(1):5-6,65.
[15]Hu M,Gasser RB.Mitochondrial genomes of parasitic nematodes-progress and perspectives[J].Trends Parasitol,2006,22(2):78-84.
[16]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.
[17]郝桂英.线粒体基因在带属绦虫分子分类中的研究进展[J].动物医学进展,2014,35(1):94-98.
[18]郝桂英,杨应东,周英姿,等.基于线粒体cox1和cytb基因对四川地区多头带绦虫的种群遗传多样性研究[J].畜牧兽医学报,2014,45(4):631-638.
[19]Zhang X,Cui J,Liu LN,et al.Genetic structure analysis of Spirometra erinaceieuropaei isolates from central and southern China[J].PLoS One,2015,10(3):e0119295.