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

doi:10.12140/j.issn.1000-7423.2024.06.016

Abstract

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

CLC Number:

R382.3

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.

Figures/Tables 5

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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