云南元江—红河流域乙型脑炎媒介蚊虫群落特征研究

发布时间：2018-01-04 08:02

本文关键词：云南元江—红河流域乙型脑炎媒介蚊虫群落特征研究　出处：《大理学院》2010年硕士论文　论文类型：学位论文

【摘要】：目的：通过对云南元江-红河流域不同地理气候环境梯度中的媒介蚊虫组成、分布和对健康人群血清学及其媒介蚊虫乙型脑炎带毒调查,探讨该流域乙型脑炎媒介蚊虫物种多样性特征及其疾病的实际流行水平。内容：(1)乙型脑炎媒介蚊虫群落特征调查：在元江-红河流域不同环境梯度(纬度和海拔)采用诱蚊灯通宵捕蚊,以观察媒介蚊虫的组成与分布等特点。(2)乙型脑炎血清学调查：在各观察点采集健康人群血清,采用血凝抑制实验检测健康人群乙型脑炎抗体IgG水平。(3)蚊虫乙型脑炎病毒自然带毒情况调查：对采集的部分蚊虫标本进行研磨,用RT-PCR、免疫荧光等实验方法进行病毒鉴定,上清液接种组织培养细胞以分离乙型脑炎病毒, 方法：(1)媒介蚊虫群落特征调查：在元江红河流域北纬23。-26。、东经100°-104。之间和海拔170-1642米之间的元江红河流域区域内,采用重点普查和抽样调查相结合的方法,选择不同经纬度(以每个纬度或经度为一水平梯度)、海拔(以每500米海拔高度为—垂直梯度)、地理气候及植被等梯度环境内的各类型生境地进行蚊虫媒介物种多样性及相关生物学的调查取样。方法采用诱蚊灯对不同环境梯度内的人、畜房通宵捕捉成蚊,以观察不同生境的蚊虫组成；通过不同环境内所获的蚊虫标本、资料和数据进行蚊虫多样性分析。(2)乙型脑炎血清学调查：在元江-红河流域的上中下游分别选取景东、新平、景谷、金平4县采集健康人血清,采用血凝抑制实验检测正常人群乙型脑炎抗体IgG水平。(3)蚊虫乙型脑炎病毒自然带毒调查：对现场调查中所捕获的成虫,经分类鉴定后分装,置于液氮罐内低温保存,带回实验室(P2或P3实验室)进行乙型脑炎病毒的检测或分离；蚊虫标本病毒检测或分离方法主要采取研磨、离心等处理后用RT-PCR、免疫荧光实验等方法进行鉴定,再通过接种组织培养C6/36细胞方法分离出乙型脑炎病毒。结果：(1)共捕获蚊虫6属37种(亚种)87875只,其中三带喙库蚊和中华按蚊属于当地的优势蚊种。结果分析发现,本研究区域蚊类物种多样性与均匀度成显著正相关(r=0.951),与生态优势度成高度负相关(r=-0.976),与纬度呈负相关(r=-0.561),与经度梯度呈正相关(r=0.411),但与海拔梯度的变化及物种多样性的关系不明显(r=0.177)。(2)景东、新平、景谷和金平共采集各县正常人血清803份,乙型脑炎病毒IgG阳性率为61.89%,其中金平县阳性率最高(75.62%),并且1-5岁儿童年龄组的抗体水平相对较低(45.21%)。(3)经采用乙型脑炎病毒引物对2属2种40批3321只雌性成蚊进行乙型脑炎病毒RT-PCR检测,结果4株为阳性,其中从三带喙库蚊中检测到2株和中华按蚊2株；后经核酸序列测定分析证实为乙型脑炎病毒；这4株病毒均对C6/36细胞能产生明显的细胞病变,经间接免疫荧光试验为乙型脑炎病毒(JEV)。结论：(1)三带喙库蚊和中华按蚊是元江-红河流域地区居民点的优势蚊种,蚊虫多样性及其丰富度与调查点所处的纬度梯度具有一定的关系。(2)该流域健康人群中流行性乙型脑炎病毒抗体水平较高,说明这些地区乙型脑炎流行程度较高。(3)乙型脑炎媒介复杂,三带喙库蚊和中华按蚊属于当地JEV的重要媒介。
[Abstract]:Objective: through the Yunnan Yuanjiang - Red River geographical and climatic gradient in mosquitoes, and distribution of virus serological investigation and mosquito vectors of Japanese encephalitis with healthy people, to explore the basin of Japanese encephalitis mosquito species diversity characteristics and actual disease prevalence levels.
Content: (1) investigation of Mosquito Community Characteristics of Japanese encephalitis in Yuanjiang - Red River environmental gradient (latitude and elevation) night mosquito mosquito lamp used to observe the characteristics of the composition and distribution of mosquitoes. (2) survey of Japanese encephalitis serological: collecting the serum of healthy individuals at each observation point, inhibition test detection of health JE antibody IgG by hemagglutination. (3) survey the natural carriers of mosquito encephalitis virus: the mosquito samples for grinding, RT-PCR, immunofluorescence test and other methods for virus identification and isolation of Japanese encephalitis virus in the supernatant were inoculated tissue culture cells,
Methods: (1) investigation of the characteristics of mosquito community media: in the Yuanjiang Red River 23.-26. north latitude, longitude 100 degrees between -104. and elevation of 170-1642 meters between the Yuanjiang Red River area, the key method of combining general and sampling survey, select the different latitude and longitude (latitude or longitude for each to a level of elevation (gradient). Every 500 meters above sea level for vertical gradient), sampling of various types of habitats and vegetation geography and climate environment in the gradient of mosquito species diversity and biology. Methods using mosquito light to different environmental gradients within the human animal, real overnight mosquito to catch mosquitoes, observe in different habitats through the different environment; the mosquito samples, data and data analysis of insect diversity. (2) survey of Japanese encephalitis serological: in the Yuanjiang Red River: selected Jingdong, new Ping, Jinping County Jinggu, 4 acquisition of healthy human serum, inhibition test to detect JE antibody level by IgG blood coagulation. (3) with natural toxins in Japanese encephalitis virus in adult mosquitoes captured in the field investigation, were identified after packing them in liquid nitrogen cryopreservation, back to the lab (P2 or P3) for Laboratory of Japanese encephalitis virus detection or isolation; mosquitoes collected virus detection and separation method mainly adopts the centrifugal grinding, etc. after treatment with RT-PCR, immunofluorescence experiments were identified by inoculating tissue culture method of C6/36 cell isolation of Japanese encephalitis virus.
Results: (1) a total of 6 genera and 37 species of mosquitoes captured (subspecies) 87875, of which three Culex and Anopheles sinensis were the dominant species. The results showed that the study area of mosquito species diversity was positively correlated with uniformity (r=0.951), and a high degree of negative ecological dominance related (r=-0.976), and negatively correlated with latitude, longitude (r=-0.561) associated with positive gradient (r=0.411), but the relationship with altitudinal variation and species diversity is not obvious (r=0.177). (2) Jing Dong, Xinping, Jinggu and Jinping counties were collected in normal human serum 803, the positive rate of hepatitis B encephalitis virus IgG was 61.89%, the positive rate of Jinping County is the highest (75.62%), and the antibody level of children aged 1-5 years old age group is relatively low (45.21%). (3) the Japanese encephalitis virus primers on 2 genera and 2 species of the 40 batch of 3321 adult female mosquitoes of Japanese encephalitis virus RT-PCR detection results of 4 strains from the positive. Three strains of Culex pipiens pallens were detected in 2 strains and 2 strains of Anopheles sinensis. They were confirmed as Japanese encephalitis virus by nucleic acid sequence analysis. These 4 viruses all had obvious cytopathic effects on C6/36 cells, and indirect immunofluorescence test was Japanese encephalitis virus (JEV).
Conclusion: (1) three Culex and Anopheles sinensis was the dominant species in Yuanjiang - Red River settlements, the latitudinal gradient mosquito diversity and richness and survey points have certain relationship. (2) encephalitis B virus antibody level of healthy population in the high prevalence in these areas Japanese encephalitis in a higher degree. (3) Japanese encephalitis complex, an important medium of three Culex and Anopheles sinensis belongs to the local JEV.

【学位授予单位】：大理学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R384

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