云南省蛋鸡源隐孢子虫鉴定及18S rRNA生物信息学分析  

Identification and analysis of 18S rRNA bioinformatics of layer-derived Cryptosporidium in Yunnan Province

作　　者：赵国洪 韩建强 葛丽红 李思远 杨锁柱 付凡 信爱国 赵津 ZHAO Guohong;HAN Jianqiang;GE Lihong;LI Siyuan;YANG Suozhu;FU Fan;XIN Aiguo;ZHAO Jin(Yuxi Agriculture Vocation-Technical College,Yuxi 653106,China;Yuxi City Key Laboratory of Livestock and Poultry Disease Detection,Yuxi 653106,China;Eshan County Huanian Town Agricultural and Rural Comprehensive Service Center,Yuxi 653202,China;Institute of Poultry Breeding and Poultry Disease,Yunnan Academy of Animal Husbandry and Veterinary Sciences,Kunming 650224,China)

机构地区：[1]玉溪农业职业技术学院,云南玉溪653106 [2]玉溪市畜禽疫病检测重点实验室,云南玉溪653106 [3]峨山县化念镇农业农村综合服务中心,云南玉溪653202 [4]云南省畜牧兽医科学院养禽与禽病研究所,昆明650224

出　　处：《黑龙江畜牧兽医》2022年第19期76-79,142,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：云南省教育厅科学研究基金项目(2021J1048);玉溪农业职业技术学院动物疫病研究中心项目(2021YZ03);信爱国专家科研工作站项目。

摘　　要：为了了解云南省规模化蛋鸡场腹泻病鸡隐孢子虫的感染情况,试验采用卢戈氏碘液和改良抗酸染色镜检、巢式PCR扩增隐孢子虫18S rRNA基因、BLAST程序测序及遗传进化分析等方法对2020年12月份从云南省多个规模化蛋鸡场采集的71份腹泻蛋鸡的粪便样品进行研究。结果表明:经卢戈氏碘液染色后可见呈棕色近似香蕉形的子孢子,经改良抗酸染色后可见呈红色近似球形的卵囊和近似香蕉形的子孢子。巢氏PCR扩增得到大小为821 bp的目的片段,将获得的虫株命名为YX202106虫株。YX202106虫株与GenBank中已公布的火鸡隐孢子虫的同源性最高,为99.14%~99.51%,且在进化树上处于同一个分支,该虫株还与能引起人畜共患的五种主要隐孢子虫——人隐孢子虫、微小隐孢子虫、火鸡隐孢子虫、猫隐孢子虫、犬隐孢子虫在进化树上构成一个大分支,具有重要的公共卫生学意义。说明云南地区蛋鸡群中存在火鸡隐孢子虫感染,应加强蛋鸡隐孢子虫病的防治。In order to understand the infection status of Cryptosporidium in diarrhea chicken of large-scale layer farms in Yunnan Province,the experiment used the methods such as Lugol’s iodine solution and modified acid-fast staining microscopy,nested PCR amplification of Crypto⁃sporidium 18S rRNA gene sequencing BLAST,and genetic evolution analysis,for the study on the fecal samples of 71 diarrheal layers collected from several large-scale layer farms in Yunnan Province in December 2020.The results showed that brownish and banana-shaped sporozoites could be seen by Lugol’s iodine solution,and red nearly spherical oocysts and banana-shaped sporozoites could be seen by modified acid-fast staining.The target fragments with a size of 821 bp were obtained by nest PCR,and the obtained strain was named YX202106 strain.The YX202106 strain had the highest homology with Cryptosporidium meleagridis published in GenBank,ranging from 99.14%to 99.51%,and was in the same branch on the evolutionary tree.The strain also formed a large branch on the evolutionary tree with the five major Cryptosporidium species that could cause zoonotic disease,including Cryptosporidium hominis,Cryptosporidium parvum,Cryptosporidium meleagridis,Cryptospo⁃ridium felis,and Cryptosporidium canis,which had important public health significance.The results suggested that there was Cryptosporidium meleagridis infection in layers in Yunnan Province,and the prevention and control of Cryptosporidium in layers should be strengthened.

关 键 词：蛋鸡 腹泻 火鸡隐孢子虫 鉴定 18S rRNA 

分 类 号：S855.9[农业科学—临床兽医学]