青藏高原东部石渠县虻类昆虫的遗传进化特征及寄生虫携带状况调查

doi:10.12140/j.issn.1000-7423.2026.02.015

中国寄生虫学与寄生虫病杂志 ›› 2026, Vol. 44 ›› Issue (2): 252-259.doi: 10.12140/j.issn.1000-7423.2026.02.015

青藏高原东部石渠县虻类昆虫的遗传进化特征及寄生虫携带状况调查

李梦晴¹(), 王旭¹, 杨扬¹^,²^,³, 薛垂召¹, 左清秋¹, 尹建海¹, 曹建平¹^,²^,³^,^*()()

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联合研究中心，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
³ 上海交通大学公共卫生学院，上海 200025

收稿日期:2026-02-04 修回日期:2026-04-13 出版日期:2026-04-30 发布日期:2026-04-22
通讯作者: * 曹建平（ORCID：0000-0002-1974-0047），男，博士，研究员，从事寄生虫病防治与研究。E-mail：caojp@chinacdc.cn
作者简介:李梦晴，女，硕士研究生，从事吸血昆虫流行病学研究。E-mail：lmq20012019@outlook.com
作者贡献
李梦晴负责论文撰写和实验操作，李梦晴、王旭、杨扬、薛垂召和左清秋负责样品采集和鉴定，王旭和杨扬参与实验操作，王旭和尹建海负责论文修改，曹建平负责论文设计、修改和审校。
基金资助:
国家自然科学基金(82404325)

Genetic evolutionary characteristics of and prevalence of parasitic infections in tabanid flies in Shiqu County, eastern Qinghai-Tibet Plateau

LI Mengqing¹(), WANG Xu¹, YANG Yang¹^,²^,³, XUE Chuizhao¹, ZUO Qingqiu¹, YIN Jianhai¹, CAO Jianping¹^,²^,³^,^*()()

¹ National Institute of Parasitic Diseases at 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 of Parasite and Vector Biology, Ministry of Health; World Health Organization Collaborating Centre for Tropical Diseases; National Center for International Research on Tropical Diseases, Ministry of Science and Technology, Shanghai, China
² School of Global Health, Chinese Center for Tropical Diseases Research, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
³ School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

Received:2026-02-04 Revised:2026-04-13 Online:2026-04-30 Published:2026-04-22
Supported by:
National Natural Science Foundation of China(82404325)

摘要/Abstract

摘要：

目的分析青藏高原东部石渠县虻类昆虫的分布特征及其潜在携带寄生虫情况，为掌握该地区虻类昆虫的分布格局及制定虻传疾病防控策略提供基础资料。方法 2023年8月和2025年8月在四川省石渠县的尼呷镇、色须镇、洛须镇，于10:00—14:00采用网兜捕捉法采集虻，提取其组织DNA。使用28S rRNA特异性引物扩增目标片段并测序，使用Geneious Prime软件拼接序列，在GenBank中进行BLAST比对以确定虫种。利用DnaSP计算单倍型多样性，DAMBE检验碱基替换饱和度，jModelTest筛选核苷酸替代模型，并用MEGA最大似然法构建系统进化树。使用通用寄生虫引物开展高通量测序，测序数据经过质量控制、双端拼接、操作分类单位（OTU）聚类等，检测虻样品中潜在寄生虫。结果共采集虻样品324份，经鉴定筛选共获得有效样品304份，鉴定为3属3种，分别为双斑瘤虻、红褐腹虻和毛肾角鹬虻。红褐腹虻共275只，为优势物种（占90.46%，275/304）；双斑瘤虻共24只，占7.89%（24/304）；毛肾角鹬虻共5只，占1.65%（5/304）。共检测到8个单倍型，其中双斑瘤虻2个单倍型，红褐腹虻和毛肾角鹬虻各3个单倍型。系统进化树分析显示，研究序列在科、属和种级水平均呈现清晰的分化结构，双斑瘤虻的2个单倍型（H3、H4）与参考序列M243434序列聚为一支，红褐腹虻的3个单倍型（H1、H6、H8）与参考序列AF238561聚为单系分支，毛肾角鹬虻的3个单倍型（H2、H5、H7）与参考序列AF238558形成高度支持的单系分支。高通量测序共获得1 499个OTU，包含2个潜在寄生虫相关OTU，分别为鼠突线虫、伊氏罗索线虫，均仅在双斑瘤虻中检出；红褐腹虻和毛肾角鹬虻中未检出寄生虫OTU。结论石渠县虻类群落以红褐腹虻为主，整体遗传多样性较低。双斑瘤虻中检测到低丰度线虫相关序列，提示其在高原虻传病原传播中的潜在生态学意义。

关键词: 吸血昆虫, 虻, 遗传进化, 青藏高原, 石渠

Abstract:

Objective To investigate the distribution characteristics of and prevalence of parasitic infections in tabanid flies in Shiqu County, eastern Qinghai-Tibet Plateau, so as to provide baseline data for understanding tabanid fly distribution patterns and formulation of tabanid-borne disease control strategies in this region. Methods Tabanid samples were collected from Nixia Town, Sexu Town, and Luoxu Town of Shiqu County, Sichuan Province using a net trapping method from 10:00 to 14:00 in August 2023 and August 2025. Genomic DNA was extracted from tabanid samples, and target fragments were amplified and sequenced using 28S rRNA-specific primers. Sequences were assembled using the Geneious Prime software, and the species was identified with the BLAST tool in the GenBank database. Haplotype diversity was calculated using the DnaSP software, and saturation of base substitutions was tested with the DAMBE software. Nucleotide substitution models were screened with the jModelTest tool, and a phylogenetic tree was generated with the maximum likelihood method using the MEGA software. High-throughput sequencing was performed with universal parasite primers, and sequencing data were subjected to quality control, paired-end assembly, operational taxonomic unit (OTU) clustering to detect potential parasites in tabanid samples. Results A total of 324 tabanid specimens were sampled, and 304 valid samples were obtained following identification and screening, which belonged to three genera and three species, including Hybomitra bimaculate (Tabanidae), Tabanus rufofrator (Tabanidae), and Symphoromyia hirta (Rhagionidae). T. rufofrator was the dominant species (275 individuals, 90.4%, 275/304), and 24 (7.89%, 24/304) H. bimaculate and 5 S. hirta (1.65%, 5/304) were identified. Totally eight haplotypes were detected, including two for H. bimaculata and three each for T. rufofrator and S. hirta. Phylogenetic analysis revealed clear differentiation structures of the study sequences at family, genus, and species levels. Two H. bimaculata haplotypes (H3 and H4) were clustered into the same clade with the reference sequence with the GenBank accession number of M243434, and three T. rufofrator haplotypes (H1, H6 and H8) were clustered into the monophyletic clade with the reference sequence with the GenBank accession number of AF238561, while three S. hirta haplotypes (H2, H5 and H7) and the reference sequence with the GenBank accession number of AF238558 generated a highly supported monophyletic clade. High-throughput sequencing yielded 1 499 OUT, which contained two potential parasite-associated OUT, including Mastophorus muris and Romanomermis iyengari, which were only present in H. bimaculate, while no parasites were detected in T. rufofrator or S. hirta. Conclusion T. rufofrator is the dominant species of the tabanid fly community in Shiqu County, with an overall low genetic diversity. Low-abundance nematode sequences are detected in H. bimaculate, suggesting the potential ecological significance in the transmission of tabanid-borne pathogens on the Qinghai-Tibet Plateau.

Key words: Blood-sucking insect, Tabanid, Genetic evolution, Qinghai-Tibet Plateau, Shiqu

中图分类号:

R384.22

李梦晴, 王旭, 杨扬, 薛垂召, 左清秋, 尹建海, 曹建平. 青藏高原东部石渠县虻类昆虫的遗传进化特征及寄生虫携带状况调查[J]. 中国寄生虫学与寄生虫病杂志, 2026, 44(2): 252-259.

LI Mengqing, WANG Xu, YANG Yang, XUE Chuizhao, ZUO Qingqiu, YIN Jianhai, CAO Jianping. Genetic evolutionary characteristics of and prevalence of parasitic infections in tabanid flies in Shiqu County, eastern Qinghai-Tibet Plateau[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2026, 44(2): 252-259.

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物种 Species	引物 Primer	引物序列（5'→3'） Primer sequence (5'→3')	产物长度/bp Product length/bp	退火温度/℃ Annealing temperature/℃
无脊椎动物 Invertebrate	JB	F-TTTTTTGGGCATCCTGAGGTTTAT	440	55.0
	JB	R-TAAAGAAAGAACATAATGAAAATG	440	55.0
阿米巴门 Amoebozoa	Amo	F-GAATTGACGGAAGGGCACAC	333~420	57.8
	Amo	R-GCCCYRTCTAAGGGCATCAC	333~420	57.8
顶复门A Apicomplexa A	Api-A	F-GACCTATCAGCTTTCGACGG	244~248	55.0
	Api-A	R-CCCTCCAATTGWTACTCTGGR	244~248	55.0
顶复门B Apicomplexa B	Api-B	F-TGYGTTTGAATACTAYAGCATGG	228~244	53.7
	Api-B	R-TCTGATCGTCTTCACTCCCTT	228~244	53.7
顶复门 Apicomplexa	Api-C	F-TTGGMCTACCGTGGCARTGA	420~427	57.5
	Api-C	R-TCAAGGCAAHWGCCTGCTT	420~427	57.5
芽囊原虫属 Blastocystis	Bla	F-TGGTCGCAAGGCTGAAACTT	303~307	58.1
	Bla	R-TTGCCTCCAGCTTCCCTACA	303~307	58.1
双滴虫目 Diplomonadida	Dip	F-RGGGACRGGTGAAATAGGATG	277~285	56.5
	Dip	R-CAAATTGAGCCGCAGACTCC	277~285	56.5
动质体门 Kinetoplastida	Kin	F-AAATTAAACCGCACGCTCCA	250~300	55.4
	Kin	R-GCAAACGATGACACCCATGA	250~300	55.4
小孢子虫门 Microsporidia	Mic	F-BCAGGTTGATTCTGCCTGACR	370~440	57.9
	Mic	R-ACCAGWCTTGCCCTCCARTT	370~440	57.9
副基体纲 Parabasalia	Par	F-TAGGCTATCACGGGTAACGG	326~364	55.3
	Par	R-GCGTCCTGATTTGTTCACAG	326~364	55.3
扁形动物门 Platyhelminthes	Pla	F-CAATTGGAGGGCAAGTCTGG	350~550	55.4
	Pla	R-TGCTTTCGCWKTAGTTTGTCTG	350~550	55.4
线虫动物门A Nematode A	Nem-A	F-CACCCGTGAGGATTGACAG	320~335	54.1
	Nem-A	R-CGATCACGGAGGATTTTCAA	320~335	54.1
线性动物门B Nematode B	Nem-B	F-CGTCATTGCTGCGGTTAAAA	380~410	56.0
	Nem-B	R-CCGTCCTTCGAACCTCTGAC	380~410	56.0
线性动物门C Nematode C	Nem-C	F-AGTGGAGCATGCGGCTTAAT	380~440	56.6
	Nem-C	R-TGCAATTCCCTRTCCCAGTC	380~440	56.6

青藏高原东部石渠县虻类昆虫的遗传进化特征及寄生虫携带状况调查

Genetic evolutionary characteristics of and prevalence of parasitic infections in tabanid flies in Shiqu County, eastern Qinghai-Tibet Plateau

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