Molecular epidemiological investigation of Babesia infection in small mammals in the central Hengduan Mountains of Yunnan Province

doi:10.12140/j.issn.1000-7423.2021.02.025

Abstract

Abstract:

To understand the situation of infection with Babesia spp. in small mammals in the middle part of mountainous area of Hengduan Mountain of Yunnan Province, small mammals were captured by trap rows and cages (in the daytime) in Jianchuan, Yunlong and Eryuan counties of the area in 2013, 2016 and 2020. The species of the captured animals was identified by morphology, and DNA was extracted from the spleen tissues. The 18S rRNA gene fragment of Babesia spp. was amplified by nested PCR. After bidirectional sequencing, the targeted bands were analyzed for homology. The phylogenetic tree was constructed by the Neighbor-Joining method. A total of 498 small mammals were captured, including 212 in Jianchuan County, 171 in Yunlong County and 115 in Eryuan county. They were morphologically identified of 27 species, belonging to 14 genera, 8 families of 4 orders. The dominant rodent species included Apodemus chevrieri, A. draco and Eothenomys miletus. The 18S rRNA gene fragment of Babesia spp. was amplified from 55 samples, with an overall positive rate of 11% (55/498). The positive rate in Jianchuan County, Yunlong County and Eryuan County was 0.5% (1/212), 31.6% (54/171) and 0 (0/115), respectively. The detection rate was relatively high in A. latronum (11/17), E. elegusis (5/10) and Soriculus leucops (2/4). The results of BLAST analysis showed that the sequences amplified in the samples were highly homologous (similarity 99.6%) and belong to the Otsu type of B. microti. There was Babesia infection in small mammals in the mid-mountainous area of Hengduan Mountain, Yunnan Province, the Babesia belonging to the Otsu type of B. microti.

Key words: Yunnan, Hengduan Mountain area, Small mammals, Babesia, Investigation

CLC Number:

R531.9

WANG Fan, BIE Shuang-shuang, ZHANG Yun, SONG Jing, DUAN Xing-de, XIONG Meng-tao, WANG Jian, JIANG Jia-fu, GAO Zi-hou, DONG Yi, DU Chun-hong. Molecular epidemiological investigation of Babesia infection in small mammals in the central Hengduan Mountains of Yunnan Province[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(2): 278-282.

Figures/Tables 4

References 19

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基因	PCR轮次	引物	引物序列（5′→3′）	预计片段大小/bp
18S rRNA 基因部分片段	第1轮	Bab1A	GTCTTAGTATAAGCTTTTATACAGCG	150
	第1轮	Bab4A	GATAGGTCAGAAACTTGAATGATACATCG
	第2轮	Bab2A	CAGTTATAGTTTATTTGATGTTCGTTTTAC
	第2轮	Bab3A	CGGCAAAGCCATGCGATTCGCTAAT
18S rRNA 基因近似全长片段	第1轮	Piro0F	GCCAGTAGTCATATGCTTGTGTTA	1 600
	第1轮	Piro6R	CTCCTTCCTYTAAGTGATAAGGTTCAC
	第2轮	Piro1F	CCATGCATGTCTWAGTAYAARCTTTTA
	第2轮	Piro5.5R	CCTYTAAGTGATAAGGTTCACAAAACTT

目	科	属	种	样本数	阳性数	阳性率/%
啮齿目Rodetia	仓鼠科Cricetidae	绒鼠属Eothenomys	大绒鼠E. miletus	98	0	0
			滇绒鼠E. eleusis	10	5	5/10
			黑腹绒鼠E. melanogaster	5	0	0
	鼠科Muridae	姬鼠属Apodemus	齐氏姬鼠A. chevrieri	162	0	0
			小林姬鼠A. sylvaticus	2	0	0
			中华姬鼠A. draco	98	34	34.7
			大耳姬鼠A. latronum	17	11	11/17
		家鼠属Rattus	斯氏家鼠R. Brunneusculus	4	1	1/4
			斑胸鼠R. yunnanensis	1	0	0
			褐家鼠R. Norvegicus	4	0	0
			黄胸鼠R. tanezumi	9	0	0
			大足鼠R. Nitidus	1	0	0
		白腹鼠属Niviventer	川西白腹鼠N. excelsior	19	1	1/19
			灰腹鼠N. eha	1	0	0
			社鼠N. confucianus	1	0	0
			梵鼠N. brahma	1	0	0
		滇攀鼠属Vernaya	滇攀鼠V. fulva	1	0	0
	松鼠科Sciuridae	花松鼠属Tamiops	隐纹花松鼠T. swinhoei	4	0	0
食虫目Insectivora	鼩鼱科Soricidae	长尾鼩鼱属Soriculus	褐鼩鼱S. caudatus	11	0	0
			印度长尾鼩鼱S. leucops	4	2	2/4
		鼩鼱属Sores	高山鼩鼱S. alpinus	5	0	0
		微尾鼩属Anourosorex	四川短尾鼩A. squamipes	5	0	0
		麝鼩属Crocidura	灰麝鼩C. attenuata	19	0	0
	猬科Erinaceidae	鼩猬属Neotetracus	中华新猬N. sinensis	4	1	1/4
	鼹科Talpidea	长吻鼩鼹属Nasillus	多齿鼩鼹N. gracilis	4	0	0
攀鼩目Scandentia	树鼩科Tupaiidae	树鼩属Tupaia	贝氏树鼩鼱T. belangeri	6	0	0
食肉目Carnivora	鼬科Mustelidae	鼬属Mustela	黄鼬M. sibirica	2	0	0
合计				498	55	11