Chines Journal of Vector Biology and Control ›› 2018, Vol. 29 ›› Issue (1): 42-46.DOI: 10.11853/j.issn.1003.8280.2018.01.011

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Species identification of Anopheles mosquitoes from the Tumen river basin in Yanbian area of Jilin province based on morphological and molecular characteristics

LI Ji-xu1, SONG Zhen-hai2, PIAO Guang-ming3   

  1. 1 Yanbian Center for Disease Control and Prevention, Yanbian 133001, Jilin Province, China;
    2 Tumen Center for Disease Control and Prevention;
    3 Longjing Center for Disease Control and Prevention
  • Received:2017-08-21 Online:2018-02-20 Published:2018-02-20
  • Supported by:
    Supported by the Aid Projects of International Infectious Disease Detection System of the National Institutes of Health, Center for Disease Control and Prevention, South Korea

吉林省延边地区图们江流域按蚊形态学及基因特征分析

李基旭1, 宋振海2, 朴光明3   

  1. 1 延边朝鲜族自治州疾病预防控制中心病媒科, 吉林 延吉 133001;
    2 图们市疾病预防控制中心, 吉林 图们 133100;
    3 龙井市疾病预防控制中心, 吉林 龙井 133400
  • 作者简介:李基旭,男,主任医师,主要从事病媒生物监测及相关传染病实验室诊断工作,Email:lijixu2006@163.com
  • 基金资助:
    韩国疾病预防控制中心国立保健研究院国际感染性疾病检测体系援助项目

Abstract: Objective To understand the species of Anopheles mosquitoes in the Tumen river basin in Yanbian area of Jilin province, and master the morphological and gene characteristics of common Anopheles mosquitoes. Methods The three border towns were chosen in the Tumen river basin in Yanbian area, Yueqing, Qushui, Kaishan in 2012-2013. From May to October every year, the black light trap was used to collect the female adult of Anopheles monthly, from which the genome DNA was extracted. The PCR gene identification technique was used to identify the Anopheles species according to the characterization of its ribosomal nucleic acid internal transcribed spacer 2 segment (rDNA ITS2) gene. The genome DNA amplification products of Anopheles were sequenced, and homology analysis with gene sequence of Anopheles in GenBank of NCBI was carried out to further determine the genetic characteristics of Anopheles mosquitoes in Tumen river basin of Yanbian area. Finally, according to the molecular identification results, reverse analyses was conducted to provide the confirmation for morphological classification. Results Five Anopheles species including An. hycranus group An. sineroides, An. kleini, An. lesteri, An. pullus, and An. belenrae were found in the study area, and the homology between their genomic DNA amplification sequences and that of Anopheles published in GenBank reached 97%-100%. The subcostal, preapical, apical fringe and fringe had obvious species specificity in morphology among An. kleini, An. lesteri, and An. pullus. Conclusion Anopheles sinensis and An. koreicus were not found in Tumen river basin of Yanbian area, An. belenrae was found for the first time in this area and became the new recorded species with An. kleini. The initial recorded species of An. anthropophagus was revised to An. lesteri. The An. yatsushiroensis was revised to An. pullus. According to the white as spots of wings as species specificity of female adults Anopheles mosquitoes, we can identify the morphology of new and related species rapidly and conveniently, such as An. kleini, An. lesteri, and An. pullus.

Key words: Anopheles, Species identification, Morphology, Internal transcription spacer, Malaria

摘要: 目的 了解吉林省延边朝鲜族自治州(延边州)图们江流域按蚊种类,掌握常见按蚊形态特征和基因特征。方法 于2012-2013年5-10月选择吉林省延边州图们市月晴镇、曲水镇和龙井市开山屯镇3个边境村,利用诱蚊灯法采集当地按蚊雌成虫,提取其基因组DNA,利用其核糖体核酸内转录间隔2区段(rDNA ITS2)基因特征,应用PCR基因鉴别技术,确定其种类。对按蚊基因组DNA扩增产物进行测序,与GenBank中按蚊序列进行同源性分析,进一步确定延边地区图们江流域按蚊基因特征。根据分子生物学鉴定结果,反向分析其形态特征。结果 在延边州图们江流域共发现赫坎按蚊种团(Anopheles hycranus group)类中华按蚊、克莱按蚊、雷氏按蚊、暗灰按蚊(近黑按蚊)和比伦按蚊,其蚊种基因组DNA扩增序列与GenBank中已发表相应按蚊序列的同源性达97%~100%。克莱按蚊、暗灰(近黑)按蚊和雷氏按蚊的亚缘脉白斑、亚尖端白斑、尖端白斑和翅5.2纵脉末端繸斑(fringe)在形态学上存在明显种特异性。结论 在延边州图们江流域未发现中华按蚊和朝鲜按蚊。比伦按蚊为该地首次发现,与已发现的克莱按蚊同为该地首次纪录种。将原纪录种嗜人按蚊订正为雷氏按蚊,原八代按蚊订正为暗灰按蚊。利用按蚊雌成虫翅白斑种特异性可对克莱按蚊、暗灰(近黑)按蚊和雷氏按蚊等新蚊种和近缘种快速、便捷地进行形态学鉴定。

关键词: 按蚊, 种类鉴定, 形态学, 内转录间隔区, 疟疾

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