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

doi:10.12140/j.issn.1000-7423.2024.01.009

Abstract

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

CLC Number:

R382.3

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.

Figures/Tables 4

Table 1

Fig. 1

Fig. 2

Table 2

Detection of E. bieneusi in dogs and cats in China

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]

Table 2

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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