云南横断山中部山区小型兽类感染巴贝虫的分子流行病学调查

doi:10.12140/j.issn.1000-7423.2021.02.025

中国寄生虫学与寄生虫病杂志 ›› 2021, Vol. 39 ›› Issue (2): 278-282.doi: 10.12140/j.issn.1000-7423.2021.02.025

• 研究简报 • 上一篇

云南横断山中部山区小型兽类感染巴贝虫的分子流行病学调查

王帆¹^,²(), 别双双¹^,², 张云², 宋静², 段兴德², 熊孟韬², 王剑², 江佳富³, 高子厚², 董毅², 杜春红²^,^*()

1 大理大学公共卫生学院,大理 671000
2 云南省地方病防治所,大理 671000
3 中国人民解放军军事科学院军事医学研究院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京 100071

收稿日期:2020-09-29 修回日期:2020-11-12 出版日期:2021-04-30 发布日期:2021-04-30
通讯作者: 杜春红
作者简介:王帆（1982-）,女,硕士研究生,从事自然疫源性疾病研究。E-mail： wangf990930@163.com
基金资助:
国家自然科学基金(81760607);国家自然科学基金(U2002219);病原微生物生物安全国家重点实验室开放基金(SKLPBS1833)

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

WANG Fan¹^,²(), BIE Shuang-shuang¹^,², ZHANG Yun², SONG Jing², DUAN Xing-de², XIONG Meng-tao², WANG Jian², JIANG Jia-fu³, GAO Zi-hou², DONG Yi², DU Chun-hong²^,^*()

1 School of Public Health, Dali University, Dali 671000, China
2 Yunnan Institute for Endemic Diseases Control and Prevention, Dali 671000, China
3 State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Science, People’s Liberation Army, Beijing 100071, China

Received:2020-09-29 Revised:2020-11-12 Online:2021-04-30 Published:2021-04-30
Contact: DU Chun-hong
Supported by:
National Natural Science Foundation of China(81760607);National Natural Science Foundation of China(U2002219);State Key Laboratory of Pathogen and Biosecurity of China(SKLPBS1833)

摘要/Abstract

摘要：

为了解云南省横断山中部山区小型兽类感染巴贝虫的情况,分别于2013、2016和2020年在云南省横断山中部山区的剑川、云龙和洱源等3个县,采用笼日法和夹线法捕获小型兽类。通过形态学进行种类鉴定,并取脾组织,提取巴贝虫DNA,巢式PCR扩增巴贝虫18S rRNA基因片段,目的条带经双向测序后,进行同源性分析,采用Neighbor-Joining法构建系统进化发育树。共捕获小型兽类498只,通过形态学鉴定,分属于4目8科14属27种,主要优势鼠种有齐氏姬鼠、中华姬鼠和大绒鼠,共有55份样品扩增出巴贝虫18S rRNA基因片段,总阳性率为11%（55/498）。剑川县、云龙县和洱源县的小型兽类巴贝虫阳性率分别为0.5%（1/212）、31.6%（54/171）和0（0/115）。巴贝虫阳性检出率比较高的小兽为：大耳姬鼠11/17,滇绒鼠5/10,印度长尾鼩鼱2/4。BLAST比对结果表明,本次检出的样品序列具有高度同源性,相似性达99.6%,属于田鼠巴贝虫的大津（Otsu）型。云南省横断山中部山区小型兽类存在巴贝虫感染,属于田鼠巴贝虫的Otsu型。

关键词: 云南, 横断山区, 小型兽类, 巴贝虫, 调查

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

中图分类号:

R531.9

王帆, 别双双, 张云, 宋静, 段兴德, 熊孟韬, 王剑, 江佳富, 高子厚, 董毅, 杜春红. 云南横断山中部山区小型兽类感染巴贝虫的分子流行病学调查[J]. 中国寄生虫学与寄生虫病杂志, 2021, 39(2): 278-282.

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.

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参考文献 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

云南横断山中部山区小型兽类感染巴贝虫的分子流行病学调查

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

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