安徽省部分地区马属动物隐孢子虫的分子检测和遗传特征

doi:10.12140/j.issn.1000-7423.2024.06.016

中国寄生虫学与寄生虫病杂志 ›› 2024, Vol. 42 ›› Issue (6): 796-801.doi: 10.12140/j.issn.1000-7423.2024.06.016

安徽省部分地区马属动物隐孢子虫的分子检测和遗传特征

王继谱(), 张玉瑾, 吴星华, 王明星, 祝玉巧, 罗辉, 高思潮, 刘欣超, 李文超^*()

安徽科技学院动物科学学院动物营养调控与健康安徽省重点实验室，安徽凤阳 233100

收稿日期:2024-04-12 修回日期:2024-06-15 出版日期:2024-12-30 发布日期:2025-01-14
通讯作者: *李文超（1979—），男，博士，教授，从事人兽共患原虫分子流行病学研究。E-mail：liwen303@126.com
作者简介:王继谱（1998—），男，硕士研究生，从事人兽共患原虫分子流行病学研究。E-mail：345318645@qq.com
基金资助:
安徽省牛羊产业技术体系(AHCYJSTX-07);安徽科技学院-潘集区肉牛产业研究院项目(AKPN2303);安徽科技学院校级稳定人才项目(dkwd201702);大学生创新创业项目(202310879038);大学生创新创业项目(S202310879065)

Molecular detection and genetic characteristics of Cryptosporidium spp. in Equus animals in some areas of Anhui Province

WANG Jipu(), ZHANG Yujin, WU Xinghua, WANG Mingxing, ZHU Yuqiao, LUO Hui, GAO Sichao, LIU Xinchao, LI Wenchao^*()

Anhui Province Key Laboratory of Animal Nutritional Regulation and Health, College of Animal Science, Anhui Science and Technology University, Fengyang 233100, Anhui, China

Received:2024-04-12 Revised:2024-06-15 Online:2024-12-30 Published:2025-01-14
Contact: *E-mail: liwen303@126.com
Supported by:
Special Fund for Anhui Agriculture Research System(AHCYJSTX-07);Program of Anhui Science and Technology University-Panji District Beef Cattle Industry Research Institute(AKPN2303);Stable Talent Program of Anhui Science and Technology University(dkwd201702);College Students’ Innovative Entrepreneurial Training Plan Program(202310879038);College Students’ Innovative Entrepreneurial Training Plan Program(S202310879065)

摘要/Abstract

摘要：

为了解安徽省马属动物隐孢子虫的感染情况及其遗传特征。2019年11月至2022年1月从安徽省滁州市、宿州市、安庆市的3个马场和阜阳市的1个驴场采集新鲜粪样。提取粪样DNA，巢式PCR扩增隐孢子虫核糖体小亚基RNA（SSU rRNA），SSU rRNA阳性的粪样巢式PCR扩增60-kDa糖蛋白（gp60）基因，巢式PCR阳性产物测序后进行序列比对，分析隐孢子虫虫种/基因型/基因亚型情况。共采集到马属动物粪样455份，隐孢子虫SSU rRNA阳性粪样7份，隐孢子虫感染率为1.5%（7/455），其中马和驴隐孢子虫感染率分别为1.9%（3/159）和1.3%（4/296）。3条马源SSU rRNA序列与隐孢子虫马基因型（MK775040、ON384432）序列一致性均达100%，gp60序列与ⅥaA15G4基因亚型（MK761060）序列一致性均达100%；4条驴源SSU rRNA序列与人隐孢子虫（KR296813）序列一致性均达99.58%，gp60序列与ⅠkA16G1基因亚型（KU200963、KU200958和KU200962）序列一致性均达100%。本研究结果显示，安徽省马属动物中存在隐孢子虫感染，感染虫种主要为隐孢子虫马基因型和人隐孢子虫。

关键词: 隐孢子虫, 马, 驴, 安徽省

Abstract:

To investigate the infection status and genetic characteristics of Cryptosporidium in Equus animals in Anhui Province, fresh fecal samples were collected from three horse farms in Chuzhou, Suzhou and Anqing, as well as one donkey farm in Fuyang between November 2019 and January 2022. DNA was extracted from the fecal samples, and nested PCR was used to amplify the Cryptosporidium small subunit ribosomal RNA (SSU rRNA). For samples that tested positive for SSU rRNA, nested PCR was further performed to amplify the 60-kDa glycoprotein (gp60) gene. Sequencing of the nested PCR positive products was conducted, followed by sequence alignment to analyze the Cryptosporidium species/genotypes/subgenotypes. A total of 455 fecal samples from Equus animals were collected, with 7 samples positive for the Cryptosporidium SSU rRNA, resulting in an infection rate of 1.5% (7/455). The infection rates for horses and donkeys were 1.9% (3/159) and 1.3% (4/296), respectively. The 3 horse-derived SSU rRNA sequences showed 100% identity with the sequences of Cryptosporidium horse genotype (MK775040, ON384432), and the gp60 sequences exhibited 100% identity with the sequence of ⅥaA15G4 subtype (MK761060). The 4 donkey-derived SSU rRNA sequences had 99.58% identity with the C. hominis (KR296813) sequence, and the gp60 sequences showed 100% identity with the ⅠkA16G1 subtype (KU200963, KU200958 and KU200962) sequences. The results of this study indicate that Cryptosporidium infections exist in Equus animals in Anhui Province, with the main species being the Cryptosporidium horse genotype and C. hominis.

Key words: Cryptosporidium, Horse, Donkey, Anhui Province

中图分类号:

R382.3

王继谱, 张玉瑾, 吴星华, 王明星, 祝玉巧, 罗辉, 高思潮, 刘欣超, 李文超. 安徽省部分地区马属动物隐孢子虫的分子检测和遗传特征[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(6): 796-801.

WANG Jipu, ZHANG Yujin, WU Xinghua, WANG Mingxing, ZHU Yuqiao, LUO Hui, GAO Sichao, LIU Xinchao, LI Wenchao. Molecular detection and genetic characteristics of Cryptosporidium spp. in Equus animals in some areas of Anhui Province[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2024, 42(6): 796-801.

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参考文献 31

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基因	引物名称	引物序列（5′→3′）	产物大小/bp
SSU rRNA	18SR	TTCTAGAGCTAATACATGCG	830
	18SF	CCCTAATCCTTCGAAACAGGA
	18SNR	GGAAGGGTTGATTTATTAGATAAAG
	18SNF	AAGGAGTAGGAAACAACCTCCA
	AL3531	ATAGTCTCCGCTGTATTC	800~850
gp60	AL3535	GGAAGGAACGATGTATCT
	AL3532	TCCGCTGTATTCTCAGCC
	AL3534	GCAGAGGAACCAGCATC

特征	粪样份数	阳性份数	感染率/%
宿主
马	159	3	1.9
驴	296	4	1.3
粪样来源
滁州市	79	2	2.5
宿州市	73	1	1.4
安庆市	7	0	0/7
阜阳市	296	4	1.4
月龄
< 12月龄	105	3	2.9
> 12月龄	350	4	1.1
合计	455	7	1.5

安徽省部分地区马属动物隐孢子虫的分子检测和遗传特征

Molecular detection and genetic characteristics of Cryptosporidium spp. in Equus animals in some areas of Anhui Province

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