青海省大通县世居牦牛犊隐孢子虫感染情况调查及虫种鉴定

doi:10.12160/j.issn.1672-5190.2020.03.021

畜牧与饲料科学 ›› 2020, Vol. 41 ›› Issue (3): 106-110.doi: 10.12160/j.issn.1672-5190.2020.03.021

青海省大通县世居牦牛犊隐孢子虫感染情况调查及虫种鉴定

杜梅卓, 张才旦卓玛, 赵志刚, 顾冬花, 李国平, 伊平昌

青海省西宁市大通县畜牧兽医站,青海西宁 810199

收稿日期:2020-03-20 出版日期:2020-05-30 发布日期:2020-06-17
通讯作者: 伊平昌（1979—）,男,高级兽医师,博士,主要研究方向为动物疾病诊断和防治。
作者简介:杜梅卓（1991—）,女,助理兽医师,主要研究方向为动物疾病诊断和防治。

Infection Status Investigation and Species Identification of Cryptosporidium in Resident Yak Calves in Datong County of Qinghai Province

DU Mei-zhuo, ZHANG Caidanzhuoma, ZHAO Zhi-gang, GU Dong-hua, LI Guo-ping, YI Ping-chang

Datong County Animal Husbandry and Veterinary Medicine Station of Xining City of Qinghai Province,Xining 810199,China

Received:2020-03-20 Online:2020-05-30 Published:2020-06-17

摘要/Abstract

摘要： 旨在调查青海省大通县部分地区世居牦牛犊隐孢子虫的感染情况及其虫种类型。在大通县宝库乡和良教乡采集了200份3~5月龄世居牦牛犊粪样进行隐孢子虫检测。所有样品经蔗糖密度梯度离心法纯化处理,采用免疫荧光试验（IFT）方法进行检测,阳性样品再用套式PCR扩增18S rDNA,并进行测序和同源性分析以供虫种鉴定。结果表明：免疫荧光试验（IFT）方法镜检出3份隐孢子虫阳性样品,阳性率为1.5%（3/200）;阳性样品经套式PCR扩增18S rDNA后,有2份隐孢子虫样品呈阳性,扩增产物长度为500 bp,测序后将序列命名为QHDTC201901和QHDTC201902;系统发育分析显示,种类鉴定发现2种隐孢子虫,分别为安氏隐孢子虫（Cryptosporidium andersoni）和牛隐孢子虫（Cryptosporidium bovis）,总感染率为1.0%（2/200）。综上提示,大通县部分地区牦牛犊存在隐孢子虫感染,感染虫种为安氏隐孢子虫和牛隐孢子虫。该调查研究为该地区牛隐孢子虫病的防控提供了临床基本数据。

关键词: 牦牛犊, 隐孢子虫, 18S rDNA, 免疫荧光试验, 套式PCR

Abstract: This study aimed to investigate the infection status and species distribution of Cryptosporidium in resident yak calves in selected areas of Datong County of Qinghai Province. A total of 200 fecal samples were collected from the 3- to 5-month-old calves in Baoku Township and Liangjiao Township of Datong County, and were subsequently detected for the presence of Cryptosporidium. After all of the samples were purified by sucrose density gradient centrifugation, immunofluorescence test （IFT） method was used to screen the presence of Cryptosporidium, and the positive samples were further confirmed by using nested-PCR assay targeting 18S rDNA. The obtained positive PCR products were sequenced and subjected to homology analysis to identify the Cryptosporidium at species level. The results showed that there were 3 samples confirmed positive for the presence of Cryptosporidium by using IFT, with the positive rate of 1.5% （3/200）; the 18S rDNA of these positive samples were amplified by nested-PCR assay, and 2 of them produced a positive product with length of 500 bp; after sequencing, the obtained sequences were designated as QHDTC201901 and QHDTC201902; phylogenetic analysis revealed that two species of Cryptosporidium were identified, Cryptosporidium andersoni （C. andersoni） and Cryptosporidium bovis （C. bovis）, with a total infection rate of 1.0% （2/200）. In conclusion, the Cryptosporidium infection in yak calves in selected areas of Datong County was observed, and the prevalent species were molecularly identified as C. andersoni and C. bovis. This study provides basic clinical data for the prevention and control of yak cryptosporidiosis in this region.

Key words: yak calves, Cryptosporidium, 18S rDNA, immunofluorescence assay, nested-PCR assay

中图分类号:

杜梅卓, 张才旦卓玛, 赵志刚, 顾冬花, 李国平, 伊平昌. 青海省大通县世居牦牛犊隐孢子虫感染情况调查及虫种鉴定[J]. 畜牧与饲料科学, 2020, 41(3): 106-110.

DU Mei-zhuo, ZHANG Caidanzhuoma, ZHAO Zhi-gang, GU Dong-hua, LI Guo-ping, YI Ping-chang. Infection Status Investigation and Species Identification of Cryptosporidium in Resident Yak Calves in Datong County of Qinghai Province[J]. Animal Husbandry and Feed Science, 2020, 41(3): 106-110.

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青海省大通县世居牦牛犊隐孢子虫感染情况调查及虫种鉴定

Infection Status Investigation and Species Identification of Cryptosporidium in Resident Yak Calves in Datong County of Qinghai Province

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