上海市宠物犬和猫毕氏肠微孢子虫和卡耶塔环孢子虫感染情况及分子特性研究

doi:10.12140/j.issn.1000-7423.2024.01.009

中国寄生虫学与寄生虫病杂志 ›› 2024, Vol. 42 ›› Issue (1): 63-68.doi: 10.12140/j.issn.1000-7423.2024.01.009

上海市宠物犬和猫毕氏肠微孢子虫和卡耶塔环孢子虫感染情况及分子特性研究

秦源¹(), 刘华¹, 王雅雪¹, 张璟¹, 苏雅馨¹, 曹建平¹^,², 沈玉娟¹^,²^,^*()

1 中国疾病预防控制中心寄生虫病预防控制所（国家热带病研究中心），传染病溯源预警与智能决策全国重点实验室，国家卫生健康委员会寄生虫病原与媒介生物学重点实验室，世界卫生组织热带病合作中心，科技部国家级热带病国际研究中心，上海 200025
2 上海交通大学医学院-国家热带病研究中心全球健康学院，上海 200025

收稿日期:2023-09-04 修回日期:2023-11-07 出版日期:2024-02-28 发布日期:2024-03-12
通讯作者: *沈玉娟（1969—），女，硕士，研究员，从事肠道原虫病原、分子流行病学、溯源等研究。E-mail：shenyj@nipd.chinacdc.cn
作者简介:秦源（1996—），女，硕士研究生，从事肠道原虫分子流行病学研究。E-mail：qy_1226@163.com
基金资助:
上海市加强公共卫生体系建设三年行动计划(2023—2025)重点学科项目(GWⅥ-11.1-09);国家自然科学基金(82372283);国家科技重大专项(2018ZX10713001-004)

Infection and molecular characterization of Enterocytozoon bieneusi and Cyclospora cayetanensis in pet dogs and cats in Shanghai

QIN Yuan¹(), LIU Hua¹, WANG Yaxue¹, ZHANG Jing¹, SU Yaxin¹, CAO Jianping¹^,², SHEN Yujuan¹^,²^,^*()

1 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention; Chinese Center for Tropical Diseases Research; National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases; Key Laboratory on Parasite and Vector Biology, Ministry of Health; WHO Collaborating Centre for Tropical Diseases; National Center for International Research on Tropical Diseases, Ministry of Science and Technology, Shanghai 200025, China
2 School of Global Health, Chinese Center for Tropical Diseases Research, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

Received:2023-09-04 Revised:2023-11-07 Online:2024-02-28 Published:2024-03-12
Contact: *E-mail: shenyj@nipd.chinacdc.cn
Supported by:
Three-Year Initiative Plan for Strengthening Public Health System Construction in Shanghai (2023—2025) Key Discipline Project(GWⅥ-11.1-09);National Natural Science Foundation of China(82372283);National Science and Technology Major Program of China(2018ZX10713001-004)

摘要/Abstract

摘要：

目的了解上海市宠物犬、猫毕氏肠微孢子虫和卡耶塔环孢子虫感染情况及分子特征，评估人兽共患传播风险。方法 2021年11月—2022年6月，采集上海市某宠物医院犬和猫的新鲜粪样，提取粪样基因组DNA，巢式PCR扩增毕氏肠微孢子虫核糖体内转录间隔区（ITS）和卡耶塔环孢子虫小亚基核糖体RNA（SSU rRNA）序列，阳性产物经双向测序后在GenBank数据库中进行BLAST比对。使用MEGA 11.0软件基于邻接法构建系统进化树。结果本研究共采集145份粪样（犬粪样99份、猫粪样46份），毕氏肠微孢子虫总阳性率为4.1%（6/145），卡耶塔环孢子虫总阳性率为4.8%（7/145）；犬的毕氏肠微孢子虫和卡耶塔环孢子虫阳性率分别为3.0%（3/99）和6.1%（6/99)，猫的毕氏肠微孢子虫和卡耶塔环孢子虫阳性率分别为6.5%（3/46）和2.2%（1/46）。6条毕氏肠微孢子虫ITS序列与人源基因型A（GenBank登录号：MK982500）的序列一致性均为100%，归在组1；7条卡耶塔环孢子虫SSU rRNA序列与人源卡耶塔环孢子虫（GenBank登录号：KJ569533）的序列一致性均为100%。系统进化树分析结果显示，毕氏肠微孢子虫分离株与已报道的人源毕氏肠微孢子虫基因型A分离株归为同一分支，卡耶塔环孢子虫的分离株与已报道的人源卡耶塔环孢子虫分离株归为同一分支。结论上海市宠物犬、猫中存在毕氏肠微孢子虫和卡耶塔环孢子虫感染，均为人兽共感染型虫株。

关键词: 毕氏肠微孢子虫, 卡耶塔环孢子虫, 宠物, 人兽共患病

Abstract:

Objective To investigate the infection and molecular characteristics of Enterocytozoon bieneusi and Cyclospora cayetanensis in pet dogs and cats in Shanghai and assess the zoonotic potential. Methods Fresh fecal samples were collected from dogs and cats at a pet hospital in Shanghai from November 2021 to June 2022. Genomic DNA was extracted from the fecal samples, and nested PCR was used to amplify the internal transcribed spacer (ITS) region of the ribosomal RNA (rRNA) of E. bieneusi and the small subunit ribosomal RNA (SSU rRNA) sequence of C. cayetanensis. The positive products were subjected to bidirectional sequencing and sequence alignment by BLAST in the GenBank database. The phylogenetic tree was constructed using MEGA 11.0 software based on the neighbor-joining method. Results A total of 145 fecal samples were collected (99 from dogs and 46 from cats). The positive rates of E. bieneusi and C. cayetanensis were 4.1% (6/145) and 4.8% (7/145), respectively. The positive rates of E. bieneusi and C. cayetanensis in dogs were 3.0% (3/99) and 6.1% (6/99), respectively, while those in cats were 6.5% (3/46) and 2.2% (1/46), respectively. The ITS sequences of 6 E. bieneusi were 100% identical to human genotype A (GenBank accession number: MK982500), and the SSU rRNA sequences of 7 C. cayetanensis were 100% identical to human C. cayetanensis (GenBank accession number: KJ569533). The results of the phylogenetic tree analysis showed that all the E. bieneusi isolates belonged to the same branch as the reported human E. bieneusi genotype A isolates and fell into group 1. All the C. cayetanensis isolates belonged to the same branch as the reported human C. cayetanensis isolates. Conclusion E. bieneusi and C. cayetanensis infections were detected in pet dogs and cats in Shanghai, all of which were caused by zoonotic parasite strains.

Key words: Enterocytozoon bieneusi, Cyclospora cayetanensis, Pets, Zoonotic disease

中图分类号:

R382.3

秦源, 刘华, 王雅雪, 张璟, 苏雅馨, 曹建平, 沈玉娟. 上海市宠物犬和猫毕氏肠微孢子虫和卡耶塔环孢子虫感染情况及分子特性研究[J]. 中国寄生虫学与寄生虫病杂志, 2024, 42(1): 63-68.

QIN Yuan, LIU Hua, WANG Yaxue, ZHANG Jing, SU Yaxin, CAO Jianping, SHEN Yujuan. Infection and molecular characterization of Enterocytozoon bieneusi and Cyclospora cayetanensis in pet dogs and cats in Shanghai[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2024, 42(1): 63-68.

图/表 4

表1

图1

图2

表2

我国犬、猫的毕氏肠微孢子虫检出情况

动物种类 Animal species	地区 Area	年份 Year	阳性率/% Positive rate/%	基因型（数量） Genotype (number)	参考文献 Reference
犬 Dog	安徽 Anhui	2013	09.3（20/215）	PtEb Ⅸ (12), EbpC (4), CHD1 (2), CHD2 (2)	[10]
	重庆 Chongqing	2013—2014	11.8（4/34）	PtEb Ⅸ (2), CD8 (2)	[11]
	广东 Guangdong	2018	22.9（149/651）	PtEb Ⅸ (56), GD1 (38), GD2 (24), D (12), EbpC (7), CD9 (4), Ⅰ (2), GD3 (2), GD4 (2), GD5 (1), GD6 (1)	[12]
	河南 Henan	2013—2014	13.9（34/244）	PtEb Ⅸ (13), O (4), EbpA (2), PigEBITS5 (1), D (3), CD3 (1), CD4 (1), CD5 (1), Macaque3 (2), Peru8 (1), Type Ⅳ (1), CD2 (1), CD6 (1), CD7 (2)	[11]
	黑龙江 Heilongjiang	-	06.7（18/267）	PtEb Ⅸ (12), NED1 (1), EbpC (1), NED2 (1), NED3 (1), D (1), NED4 (1)	[13]
	吉林 Jilin	-	07.8（2/26）	CHN5 (1), CHN6 (1)	[14]
	陕西 Shaanxi	2013—2014	20.0（6/30）	PtEb Ⅸ (2), EbpC (1), CD8 (2), CD9 (1)	[11]
	上海 Shanghai	2011—2014	06.0（29/485）	PtEb Ⅸ (28), D (1)	[1]
	四川 Sichuan	2013—2014	25.0（10/40）	PtEb Ⅸ (9), CD1 (1)	[11]
	四川 Sichuan	2019—2020	18.8（136/724）	CD9 (92), PtEb Ⅸ (41), Type Ⅳ (1), SCD-1 (1), SCD-2 (1)	[15]
	云南 Yunan	-	08.0（21/262）	PtEb Ⅸ (18), CD9 (1), DgEb Ⅰ (1), DgEb Ⅱ (1)	[16]
	浙江 Zhejiang	2013	08.0（27/315）	PtEb Ⅸ (16), EbpC (4), CHD1 (2), CHD2 (2), CHD3 (3)	[10]
猫 Cat	广东 Guangdong	2018	20.3（79/389）	PtEb Ⅸ (25), GD1 (16), D (12), GD2 (10), Type Ⅳ (6), EbpC (5), GC1(3), CD9 (2)	[12]
	河南 Henan	2013—2014	11.5（11/96）	D (3), BEB6 (2), Ⅰ (1), PtEb Ⅸ (1), CC1 (1), CC2 (1), CC3 (1), CC4 (1)	[11]
	黑龙江 Heilongjiang	-	05.8（3/52）	D (2), Type Ⅳ (1)	[13]
	上海 Shanghai	2011—2014	05.6（9/160）	Type Ⅳ (5), D (4)	[1]
	四川 Sichuan	2019—2020	14.1（22/156）	CD9 (11), Type Ⅳ (6), D (4), PtEb Ⅸ (1)	[15]
	云南 Yunnan	-	02.3（4/171）	Type Ⅳ (3), CtEb Ⅰ (1)	[16]
	浙江 Zhejiang	2015	02.5（2/81）	Type Ⅳ (1), D (1)	[10]

表2

参考文献 21

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肠道原虫 Intestinal protozoa	位点 Locus	引物名称 Primer name	引物序列（5'→3'） Primer sequence (5'→3')	产物大小/bp Product size/bp
毕氏肠微孢子虫 E. bieneusi	ITS	Eb-F1	TTCAGATGGTCATAGGGATG	0 410
		Eb-R1	ATTAGAGCATTCCGTGAGG	0 410
		Eb-F2	TCGGCTCTGAATATCTATGG	0 392
		Eb-R2	ATTCTTTCGCGCTCGTC	0 392
卡耶塔环孢子虫 C. cayetanensis	SSU rRNA	Cycl1	AATGTA AAACCCTTCCAGAGTAAC	1 747
		Cycl2	GCAATAATCTATCCCCAT CACG	1 747
		Cycl3	AATTCCAGCTCCAATAGTGTA T	0 500
		Cycl4	CAGGAGAAGCCAAGGTAGGCRTTT	0 500

上海市宠物犬和猫毕氏肠微孢子虫和卡耶塔环孢子虫感染情况及分子特性研究

Infection and molecular characterization of Enterocytozoon bieneusi and Cyclospora cayetanensis in pet dogs and cats in Shanghai

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