昆明尖音库蚊复合组种类的调查与鉴定

Investigation and Identification of Species of the Culex pipiens Complex in Kunming

【目的】对云南昆明尖音库蚊复合组蚊虫的种类进行调查和鉴定,为蚊媒传染病的防控提供依据。【方法】选取于2022年6月和2023年5月采集于昆明金殿青龙山和黄土坡的尖音库蚊复合组雄虫进行形态学和分子生物学鉴定。通过体视显微镜观察蚊虫的形态特征,并切下雄蚊尾器进行观察,根据形态特征鉴定蚊虫种类;扩增蚊虫的线粒体细胞色素C氧化酶亚基Ⅰ(COⅠ)基因和乙酰胆碱酯酶基因2(AChE2),对基因序列进行分析。【结果】金殿青龙山采集的蚊虫阳茎中叶末端尖削,内叶外伸部分呈宽叶状;黄土坡采集的蚊虫阳茎中叶末端平齐,阳茎内叶呈刀叶状。基因序列相似性分析结果显示,采集蚊虫的COⅠ基因无显著差异;AChE 2基因将蚊虫区分为两种,金殿青龙山采集蚊虫与致倦库蚊相似性较高,相似性最高为98.1%~100%,黄土坡采集蚊虫与尖音库蚊指名亚种相似性较高,相似性最高为99.7%。COⅠ基因系统发育分析结果显示,6只蚊虫与致倦库蚊、尖音库蚊、骚扰库蚊和淡色库蚊处于同一进化分支,没有明显的遗传进化差异;AChE 2基因系统发育分析结果显示,金殿青龙山采集的蚊虫与致倦库蚊聚为一簇,黄土坡采集的蚊虫与尖音库蚊聚为一簇,分别处于不同进化分支。【结论】本研究采集到2种尖音库蚊复合组蚊虫。金殿青龙山采集蚊虫为致倦库蚊,黄土坡采集蚊虫为尖音库蚊指名亚种。研究表明中国昆明等南方地区存在尖音库蚊指名亚种,应加强西尼罗病毒等有关病毒的监测。

【Objective】The experiment was aimed to investigate and identify the species of Culex pipiens complex in Kunming,Yunnan,and provide data for the prevention and control of mosquito borne infectious diseases.【Method】Morphological and molecular biological identification were conducted to the male Culex pipiens complex collected from Qinglong mountain next to the Jindian and Huangtupo,Kunming,in June 2022 and May 2023.The morphological characteristics of mosquitoes were observed under stereomicroscope,and the male mosquito phallosomes was cut off for observation,and the mosquito species were identified according to the morphological characteristics.The mitochondrial cytochrome C oxidase subunitⅠ(COⅠ)gene and acetylcholinesterase gene 2(AChE2)of mosquitoes were amplified and their sequences were analyzed.【Result】Morphology showed that the middle end of the phallosomes of mosquitoes collected from Qinglong mountain next to the Jingdian was pointed,and the extension part of the inner leaf was broad leaf.The middle end of the phallosomes of mosquitoes collected from Huangtupo of Kunming is flush,and the inner leaf of the stem was blade like.The results of gene sequence similarity analysis showed that there was no significant difference in the COⅠgene of the collected mosquitoes.The AChE 2 gene distinguished mosquitoes into two types.Mosquitoes collected from Qinglong mountain next to the Jingdian had high similarity with Culex pipiens quinquefasciatus,with the highest similarity ranging from 98.1%to 100%.Mosquitoes collected from the Huangtuppo had high similarity with Culex pipiens pipiens,with the highest similarity of 99.7%.The results of COⅠgene phylogenetic analysis showed that the six mosquitoes were in the same evolutionary branch as Culex quinquefasciatus,Culex pipiens,Culex pipiens molestus,and Culex pipiens pallens,without significant genetic evolutionary differences.The phylogenetic analysis of AChE 2 gene showed that mosquitoes collected from Qinglong Mountain next to the Jingdian were clustered with Culex quinquefasciatus,while mosquitoes collected from Huangtupo were clustered with Culex pipiens,located in different evolutionary branches.【Conclusion】Two kinds of the Culex pipiens complex were collected.The mosquitoes collected from Qinglong mountain next to the Jingdian were Culex pipiens quinquefasciatus,while the mosquitoes collected from the Huangtupo were Culex pipiens pipiens.Research had shown the discovery of Culex pipiens pipiens in Southern regions such as Kunming in China,and the monitoring of West Nile virus and related viruses should be strengthened.