中国寄生虫学与寄生虫病杂志 ›› 2011, Vol. 29 ›› Issue (5): 5-348-352.

• 论著 • 上一篇    下一篇

浙江省金华地区并殖吸虫自然宿主调查及虫种鉴定

 楼宏强, 胡野, 金耀建, 余新图, 王岚, 何旭英, 屠平光   

  1. 1 金华职业技术学院人畜共患病研究所,金华 321007;2 金华市疾病预防控制中心,金华 321002
  • 出版日期:2011-10-30 发布日期:2012-09-27

Investigation on the Hosts with Natural Paragonimus Infection and Species Identification in Jinhua Prefecture of Zhejiang Province

LOU  Hong-Jiang, HU  Ye, JIN  Yao-Jian, TU  Xin-Tu, WANG  Lan, HE  Xu-Ying, TU  Ping-Guang   

  1. 1 Institute of Zoonosis,Jinhua Polytechnic,Jinhua 321007,China;2 Jinhua City Center for Disease Control and Prevention,Jinhua 321002,China
  • Online:2011-10-30 Published:2012-09-27

摘要: 目的  调查浙江省金华地区并殖吸虫自然中间宿主和终宿主感染情况,并确定并殖吸虫虫种地位。  方法  采用整群随机抽样法,在金华市的9个区(县)中,各随机抽取3个乡镇(街道),共27个调查点。现场采集标本,剖检淡水螺类,检查并殖吸虫尾蚴感染情况。以双筛水洗法检查溪蟹并殖吸虫囊蚴感染情况。从囊蚴检查阳性的调查点收集猫、狗和溪边山坑的流浪猫粪便,以水洗沉淀法检查并殖吸虫虫卵。用分离自溪蟹的并殖吸虫囊蚴人工感染家犬获取成虫。测量尾蚴、囊蚴、虫卵和成虫的大小。提取成虫基因组DNA,进行PCR扩增并殖吸虫线粒体细胞色素C氧化酶亚单位(COI)和核糖体DNA第二间区(ITS2)的基因,测序后用BoiEdit软件分析其与其他11株并殖吸虫的同源性,利用MEGA软件构建种系发生树。 结果  金华市婺城区沙畈乡和琅琊镇与武义县白姆乡均发现并殖吸虫,第一、二中间宿主分别是放逸短沟蜷和浙江华溪蟹。婺城区沙畈乡的螺类和溪蟹的感染率分别为0.2%(2/1 088)和76.7%(46/60),溪蟹的感染指数为2.0。琅琊镇的螺类和溪蟹的感染率分别为0.1%(1/1 683)和53.0%(32/60),溪蟹的感染指数为0.9。武义县白姆乡的螺类和溪蟹的感染率分别为0(0/575)和30.0%(18/60),溪蟹的感染指数为0.1。沙畈乡和琅琊镇各在1份流浪猫粪便中检出并殖吸虫虫卵,检出率分别为8.3%(1/12)和0.6%(1/17)。金华市并殖吸虫尾蚴、囊蚴、虫卵和成虫的大小以及形态与卫氏并殖吸虫基本一致。PCR结果显示,COI和ITS2片段大小分别为390 bp和363 bp,与GenBank中已报道的11株卫氏并殖吸虫的同源性分别为88.2%~98.2%和86.5%~88.1%。在种系发生树上,金华地区的并殖吸虫位于福建闽清和日本Mie与Chiba地理株之间,与前者最为接近。  结论  浙江省金华地区存在自然感染并殖吸虫的螺、蟹和家猫宿主,虫种为卫氏并殖吸虫,在进化和亲缘关系上与福建闽清卫氏并殖吸虫非常接近。

关键词: 卫氏并殖吸虫, 宿主, 自然感染

Abstract: Objective   To investigate the natural hosts infected with Paragonimus sp. and identify the species of the parasite in selected counties/districts of Jinhua prefecture in Zhejiang Province.  Methods   Three townships/towns were randomly sampled from each of the 9 counties/districts in Jinhua as pilot spots for the survey. Fresh-water snails were collected from the fields for examining cercariae. Crabs were collected and detected for metacercariae by routine technique and the metacercariae were fed to dogs purchased in areas free from paragonimiasis. Fecal materials of dogs and cats around the villages and streams where crabs were found infected were collected for examining eggs. The artificially infected dogs were sacrificed 55 d after infection to receive adult worms. The size of cercariae, metacercariae, eggs and adult worms was measured. After the DNA of the adult worm was extracted,PCR was used to amplify the COI gene and ITS2 gene of the mitochondria from the worms. Homology with relative strains/isolates was analyzed and phylo-genetic tree constructed.  Results   The survey demonstrated that the snail Semisulcospira libertina and the crab Sinopotamon chekiangense served as the first and second intermediate hosts respectively. Natural infection was found in Wucheng District with an infection rate of 0.2% (2/1 088)in snails and 76.7%(46/60)in crabs in Shafan township, and an infection index(II)of 2.0 in crabs, 0.1% (1/1 683)in snails and 53.0%(46/60)in crabs with an II of 0.9 in Langya town. The infection rate was 0(0/575)in snails and 30.0%(18/60)in crabs with an II of 0.1 in Baimu township of Wuyi County. Paragonimus eggs were detected in feces of stray cats with a positive rate of 8.3%(1/12)in Shafan and 0.6%(1/17)in Langya. The size and morphology of the cercariae, metacercariae, eggs and adult worms were similar to those of Paragonimus westermani. The sequences of the COI and ITS2 genes were with 390 bp and 363 bp respectively, indicating a homology of 88.2%-98.2% and 86.5%-88.1% to the 11 strains/isolates of P. westermani recorded in the GenBank. The phylogenetic trees revealed that the parasite in Jinhua was in between the Minchin(Minqing, Fujian)strain and the Japanese Mie and Chiba strains.  Conclusion  Natural paragonimus infection has been detected in snails, crabs and cats in Jinhua. The parasite has been identified as P. westermani which is phylogenetically close to those reported from Fujian Province and from Mie and Chiba of Japan.

Key words: Paragonimus westermani, Host, Natural infection