Larval Mosquito Feeding and Microbial Interactions

这项研究的指导原则是，从容器栖息地产生成年蚊子与有机物的微生物转化和微生物生物量的产生密切相关。这些关系通过微生物群落在不同的幼虫栖息地对蚊子幼虫的生长施加了根本的限制，并促进了蚊子的生长。有机物质的转化率被认为是由基质的质量和对微生物和幼虫生长的刺激所调节的。我们的长期目标是勾勒出蚊子生产的制约因素，确定资源的利用效率，并制定一个现实的幼虫生长模式。虽然我们把重点放在蚊子三色伊蚊和入侵树洞的日本伊蚊作为模型生态系统进行研究，但现在我们的研究定位于探索与登革热病毒媒介埃及伊蚊的相似关系。几个具体目标提供了研究框架。我们将使用我们的黄杆菌木聚糖酶基因结构来刺激聚合物酶的降解，并测量蚊子生长速度的差异，并将使用元基因组学来研究营养模拟/微生物群落功能关系，然后使用特定的引物来利用高通量测序和qPCR方法。氮被认为是促进入侵蚊种竞争优势的关键限制营养物质。我们将提供不同形式的氮，并使用同位素标记的化合物来衡量相对同化效率。幼虫的饲料中的氮素会有所不同。埃及伊蚊以确定低氮对登革病毒媒介能力的影响。一种强表达芽孢杆菌来源的杀幼虫毒素的黄杆菌结构将被评估它在天然树洞的实验室模型中的稳定整合和持久性，用qPCR和荧光记者追踪，并量化幼虫的死亡率。同样，还将研究蚊子致病植霉菌菌株的持久性。最后，我们将分析出现的蚊子的稳定同位素浓缩特征，以溯源埃及伊蚊对不同幼虫栖息地的跟踪，并量化幼虫资源利用。

The guiding principle of this research is that adult mosquito production from container habitats is intimately linked to microbial transformations of organic matter and the production of microbial biomass. These relationships impose fundamental constraints on, and facilitate, larval mosquito growth by microbial communities in a variety of larval habitats. Rates of conversion of organic matter are postulated to be modulated by quality of substrates and governed by stimulation of microbial and larval growth. Our long-term goals are to delineate the constraints on mosquito production, to identify the efficiency of utilization of resources, and to elaborate a realistic model of larval growth. Although we have focused on the mosquito Aedes triseriatus and the invasive species Aedes japonicus into water-filled tree holes as a model ecosystem for study, we have now positioned our research to explore analogous relationships with the dengue virus vector Aedes aegypti. Several specific aims provide the research framework. We will stimulate polymer enzyme degradation with our Flavobacterium xylanase genetic constructs and measure differences in mosquito growth rates, and will employ metagenomics for nutrient-simulation/microbial community function relationships generally, and then use specific primers to utilize high throughput sequencing and qPCR methods. Nitrogen is identified as a key limiting nutrient promoting competitive advantage of invasive mosquito species. We will provision nitrogen in different forms and use isotope-labeled compounds to measure relative assimilation efficiency. Nitrogen will be varied in diets of larval Ae. aegypti to determine the influence of low nitrogen on dengue virus vector competence. A Flavobacterium construct strongly expressing larvicidal toxins of Bacillus origin will be evaluated for its stable integration and persistence into laboratory models of natural tree holes, tracked with qPCR and fluorescent reporters, and mortality rates of larvae quantified. Persistence of mosquito-pathogenic Phytopythium strains will similarly be studied. Lastly, we will analyze stable isotope enrichment signatures of emerged mosquitoes to source-track Aedes aegypti to diverse larval habitats and quantify larval resource utilization.