中华按蚊和嗜人按蚊对间日疟原虫的易感性研究

发布时间：2018-03-27 17:44

本文选题：中华按蚊　切入点：嗜人按蚊　出处：《江苏省血吸虫病防治研究所》2007年硕士论文

【摘要】： 疟疾是联合国千年发展目标中重点防控的三种传染病之一,疟疾和艾滋病、结核病一起被世界卫生组织列为全球急需控制的三大公共卫生问题。在感染人类的四种疟原虫中,以恶性疟和间日疟分布最为广泛,尽管间日疟的致病性没有恶性疟强,但由于其分布范围广、易复发等特点,使得间日疟的控制比恶性疟更为困难。由于受到多种因素的影响,我国中部地区近几年的间日疟疫情明显回升,连续出现小暴发点和局部暴发流行。嗜人按蚊和中华按蚊被认为是我国分布范围最广的疟疾传播媒介,也是我国间日疟传播的主要媒介。关于中华按蚊和嗜人按蚊对间日疟原虫的易感性研究以及结合媒介生物学特性进行的传疟作用和传疟地位的评定一直以来都是疟疾传播媒介研究中的热点。按蚊对疟原虫易感性研究最直接的方法就是将疟原虫感染按蚊,而其中应用最广泛的就是体外人工膜饲血感染系统。本研究的目的是比较我国的主要传疟媒介----中华按蚊和嗜人按蚊对间日疟原虫的易感性,并评价其在我国中部间日疟流行地区疟疾传播过程中的作用。为不同媒介按蚊疟疾流行地区的疫情控制提供科学依据。研究包括三个部分: 第一部分:按蚊实验室体外人工膜饲血感染系统的建立目的:通过改进新引进的实验室按蚊体外人工膜饲血设备和技术,建立了适合我国开展疟疾传播媒介对疟原虫易感性实验研究需要的新的实验室按蚊体外人工膜饲感染系统。方法:利用引进的按蚊体外人工膜饲设备,在实验室用嗜人按蚊江苏株为实验蚊种,以正常人全血为饲血样本,对影响按蚊吸血的多种因素进行实验研究和比较。结果: 1.羽化后3-8日龄间的按蚊的吸血率无显著差异,考虑感染按蚊的寿命,选择3-4日龄的按蚊进行人工感染更合适。 2.虽然饥饿时间在8-24小时和24小时以上的按蚊的吸血率存在显著差异,但综合考虑按蚊吸血率和死亡率的相互关系,选择按蚊饲血前饥饿时间8-12小时为宜。 3.嗜人按蚊在夜晚的膜饲吸血率显著高于白天,但在白天用黑布覆盖蚊笼模似夜晚和不覆盖黑布的膜饲吸血率无显著差异,蚊笼大小对膜饲吸血率也无显著差异。第二部分:中华按蚊和嗜人按蚊对间日疟原虫易感性的研究目的:利用新建立的按蚊体外人工膜饲感染系统,进行中华按蚊和嗜人按蚊对间日疟原虫的易感性实验研究。方法:分别用中华按蚊江苏实验株和现场F1株以及江苏、辽宁和广东珠海的嗜人按蚊,利用新建立的体外人工感染系统,在实验室配对体外人工感染在我国中部疟疾流行地区采集的间日疟患者血样。结果: 1.中华按蚊江苏实验株和现场F1株共配对感染10批次,两者的卵囊和子孢子感染率及其阳性蚊比均无显著差异。 2.江苏实验株中华按蚊和嗜人按蚊共配对感染142批次,两者的卵囊和子孢子感染率及其阳性蚊比均无显著差异;虽然中华按蚊的卵囊感染度高于嗜人按蚊,但两者的子孢子感染度无显著差异。 3.江苏、广东、辽宁嗜人按蚊共配对感染35批次,三地区嗜人按蚊间的卵囊和子孢子感染率、及其阳性蚊比均无显著差异。第三部分:影响间日疟原虫在按蚊体内发育的因素分析目的:比较不同因素对按蚊实验室体外人工感染间日疟原虫结果的影响。方法:分别比较中华按蚊和嗜人按蚊对发作间歇期和临床发作期间日疟原虫的易感性;分析配子体密度、配子体的雌雄比与中华按蚊和嗜人按蚊卵囊、子孢子感染的关联;判断中华按蚊和嗜人按蚊的卵囊感染度与子孢子发育的相关性。结果: 1.中华按蚊对临床间歇期间日疟原虫的卵囊阳性率、卵囊阳性蚊比率、子孢子阳性蚊比率和子孢子感染度均高于临床发作期间日疟原虫。 2.嗜人按蚊对临床间歇期间日疟原虫的卵囊阳性蚊比率、子孢子阳性率和子孢子阳性蚊比率均显著高于临床发作期间日疟原虫。 3.中华按蚊和嗜人按蚊的卵囊和子孢子感染率和感染度均与配子体的密度有相关关系,而与配子体的雌雄比例没有相关关系。 4.中华按蚊和嗜人按蚊的卵囊感染度越高,其子孢子感染几率和感染度就越高。研究结论 1.新建立的实验室按蚊体外人工膜饲感染系统适合于我国用于开展疟疾传播媒介对疟原虫易感性实验研究的需要,在按蚊体外人工膜饲感染实验操作的标准化和规范化方面有很大提高,为我国不同实验室间疟疾传播媒介对疟原虫易感性实验研究数据的相互比较和实验结果的比较分析打下基础。 2.中华按蚊实验株可替代现场捕获的中华按蚊F1代用于中华按蚊对间日疟原虫易感性的实验研究。 3.辽宁和广东横琴岛新发现的嗜人按蚊对间日疟原虫易感,其对间日疟原虫的易感性与我国中部的嗜人按蚊无显著差异。 4.新发现中华按蚊对当前我国中部疟疾流行区的间日疟原虫易感性与嗜人按蚊易感性类似,有必要进一步开展不同基因型间日疟原虫对中华按蚊易感性的研究,为在以中华按蚊为主要媒介地区修订和制定防治策略和措施提供参考依据。 5.在按蚊体外人工膜饲感染的实验研究中,不同临床发作期的间日疟原虫血样本、疟原虫配子体密度与按蚊卵囊和子孢子感染率和感染度的高低有一定的关系,但与疟原虫配子体的雌雄比例无关。创新点: 1.建立了适合我国传播媒介对疟原虫易感性研究所需的新的实验室按蚊体外人工膜饲感染系统。 2.首次通过实验证实辽宁和广东横琴岛新发现的嗜人按蚊对间日疟原虫易感,且其易感性与我国中部的嗜人按蚊无显著差异。 3.新发现中华按蚊对当前我国中部疟疾流行区的间日疟原虫易感性与嗜人按蚊易感性相当。
[Abstract]:Malaria is one of the three major infectious diseases prevention and control of the UN Millennium Development Goals, malaria and AIDS, tuberculosis is the WHO listed as the three major global public health problems in urgent need of control. In four species of human Plasmodium parasites, Plasmodium falciparum and Plasmodium vivax in the most widely distributed, although the pathogenicity of vivax malaria no malignant malaria, but because of its wide distribution, so easy to relapse, the vivax malaria control is more difficult than malignant malaria. As a result of many factors, in recent years of vivax malaria in the middle area of China rebounded significantly, continuous small outbreak and local outbreak.
Anopheles anthropophagus and Anopheles sinensis were considered malaria vectors of China's most widely distributed, and it is the main medium in China. The evaluation on Plasmodium vivax transmission susceptibility of Anopheles sinensis and Anopheles anthropophagus to Plasmodium vivax and combined with the biological characteristics of the media of malaria and malaria transmission status is always a hot study on malaria vectors in Anopheles on Plasmodium susceptibility. The most direct way is to Anopheles Plasmodium infection, which is the most widely used artificial membrane feeding in vitro blood infection system.
The purpose of this study is to compare the main malaria vector, Anopheles sinensis and Anopheles anthropophagus to Plasmodium vivax and evaluate its susceptibility to malaria transmission in Central China vivax malaria epidemic areas in the role. To provide the scientific basis for malaria endemic areas of different Anopheles outbreak control. The research includes three parts:
First part: establishment of an in vitro artificial membrane feeding system for Anopheles mosquitoes
Objective: to establish a new laboratory Anopheles mosquito infection system in vitro by improving the new artificial Anopheles laboratory blood feeding equipment and technology.
Methods: the Anopheles sinensis artificial membrane feeding device was introduced in the laboratory. The mosquito was selected as an experimental mosquito species in the laboratory with Anopheles sinensis Jiangsu strain. Blood samples from healthy people were taken as blood samples. The factors influencing the blood sucking of Anopheles sinensis were studied and compared.
Result:
The blood sucking rate of Anopheles Anopheles at 3-8 days after 1. emergence was not significantly different. Considering the life span of Anopheles, it was more suitable for the Anopheles Anopheles at 3-4 days of age to choose artificial infection.
2., although there are significant differences in the blood sucking rate of Anopheles between 8-24 hours and 24 hours, considering the relationship between the rate of blood loss and mortality of Anopheles, choosing Anopheles for 8-12 hours before feeding blood is appropriate.
3. Anopheles anthropophagus in the blood sucking membrane feeding rate was significantly higher than that of the night during the day, but in the daytime black night and die like mosquito cage cover does not cover the black blood membrane feeding was no significant difference between the size of the cage, mosquito membrane feeding blood rate also showed no significant difference.
The second part: the study on the susceptibility of Anopheles sinensis and Anopheles Anopheles to Plasmodium vivax
Objective: To study the susceptibility test of Anopheles sinensis and Anopheles Anopheles Anopheles Anopheles and Anopheles sinensis to Plasmodium vivax using the newly established artificial membrane feeding system of Anopheles Anopheles.
Methods: using experimental strains of Anopheles sinensis in Jiangsu and F1 strains as well as Jiangsu, Liaoning and Guangdong Zhuhai anthropophagus, using the new system in the laboratory in vitro artificial infection, paired in vitro artificial infection in Central China malaria endemic areas of blood samples of vivax malaria patients.
Result:
1. the Jiangsu experimental and F1 strains of Anopheles sinensis were infected with 10 batches, and there was no significant difference in the rate of oocyst and subspore infection and the ratio of the positive mosquito.
2. there were 142 batches of Anopheles sinensis and Anopheles sinensis co infected in Jiangsu experimental group. There was no significant difference in the infection rate of oocysts and sporozoites and the proportion of positive mosquitoes between them. Although the infection rate of Anopheles sinensis was higher than that of Anopheles sinensis, there was no significant difference in their sporozoites infection.
3., there were 35 batches of Anopheles mosquitoes co infected in Jiangsu, Guangdong and Liaoning. There was no significant difference in the infection rate of oocysts and sporozoites between three regions and the proportion of positive mosquitoes.
The third part: analysis of factors affecting the development of Plasmodium vivax in Anopheles Anopheles
Objective: To compare the effects of different factors on the results of an in vitro artificial infection of Plasmodium vivax in the laboratory of Anopheles Anopheles.
Methods: comparison of Anopheles sinensis and Anopheles anthropophagus to interictal and clinical seizures during the period of susceptibility to Plasmodium vivax; analysis of gametocyte density, the ratio of female and male gametophyte and Anopheles sinensis and Anopheles anthropophagus oocyst, sporozoite infection associated; judge of Anopheles sinensis and addicted person according to mosquito oocysts and sporozoites of correlation degree development.
Result:
1., the positive rate of oocysts, the positive rate of oocysts, the ratio of sporozoite positive mosquito and sporozoite infection in Anopheles sinensis during the clinical intermission period were all higher than those in the clinical outbreak period.
2. the Anopheles Anopheles showed a positive rate of sporocyst positive sporozoites and sporozoite positive mosquitoes during clinical intermission.
3. the infection rate and infection degree of oocysts and sporozoites of Anopheles sinensis and Anopheles sinensis were all related to the density of gametophyte, but not related to the sex ratio of gametophyte.
4. the higher the oocyst infection of Anopheles sinensis and Anopheles Anopheles, the higher the incidence and infection of the sporozoites.
research conclusion
The laboratory in vitro artificial membrane blood feeding 1. new infection system suitable for our country to carry out the need for experimental study of malaria vectors susceptibility to Plasmodium, has greatly improved in laboratory in vitro artificial membrane feeding experiment operation standardization, comparative analysis for our different laboratory malaria vectors for malaria susceptibility the comparison between experimental data and experimental results lay the foundation.
2. Anopheles sinensis experimental strain can be used as a substitute for the experimental study on the susceptibility of Anopheles sinensis to Anopheles sinensis by the F1 generation of Anopheles sinensis captured on the spot.
3., Anopheles sinensis, newly discovered in Hengqin island of Liaoning and Guangdong, is susceptible to Plasmodium vivax, and its susceptibility to Plasmodium vivax is not significantly different from that of Anopheles sinensis in Central China.
The newly discovered 4. Anopheles sinensis in central part of China in malaria endemic area of Plasmodium vivax and susceptibility of Anopheles anthropophagus susceptibility is similar, it is necessary to carry out further research on different genotypes of Plasmodium vivax in Anopheles susceptibility of Anopheles sinensis was the main media in revision and to provide reference for the prevention and control measures.
5., in the experimental study of Anopheles in vitro artificial membrane feeding, the density of Plasmodium vivax blood samples and malaria gametophyte density at different clinical stages were related to the infection rate and infection degree of Anopheles mosquitoes, oocysts and sporozoites, but not related to the sex ratio of Plasmodium gametophyte.
Innovation point:
1. a new laboratory artificial membrane feeding system for Anopheles mosquito in vitro was established for the study of susceptibility to Plasmodium malaria in China.
2., for the first time, it is confirmed experimentally that the newly discovered Anopheles sinensis in Hengqin island of Liaoning and Guangdong is susceptible to Plasmodium vivax, and its susceptibility is not significantly different from that of Anopheles sinensis in Central China.
3. the susceptibility of Anopheles sinensis to Plasmodium vivax in the current malaria endemic area of central China is new to the susceptibility of Anopheles Anopheles to Anopheles Anopheles.

【学位授予单位】：江苏省血吸虫病防治研究所
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R383

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