Larval Mosquito Feeding and Microbial Interactions

本研究的指导原则是，容器生境的成蚊生产与微生物转化有机质和微生物生物量的生产密切相关。这些关系从根本上限制并促进了微生物群落在各种幼虫栖息地中蚊子幼虫的生长。有机物的转化率被认为是由基质的质量和微生物和幼虫生长的刺激来调节的。我们的长期目标是描述蚊子生产的限制，确定资源利用的效率，并制定一个现实的幼虫生长模型。虽然我们的研究重点是三体伊蚊和入侵物种日本伊蚊在充满水的树洞中作为模型生态系统进行研究，但我们现在已经将我们的研究定位于探索与登革热病毒载体埃及伊蚊的类似关系。几个具体目标提供了研究框架。我们将利用黄杆菌木聚糖酶基因构建体刺激聚合物酶降解，测量蚊子生长速率的差异，并将利用宏基因组学研究营养模拟/微生物群落功能关系，然后使用特定引物利用高通量测序和qPCR方法。氮被认为是促进入侵蚊种竞争优势的关键限制性养分。我们将提供不同形式的氮，并使用同位素标记的化合物来测量相对同化效率。伊蚊幼虫的饮食中氮含量会发生变化。测定低氮对埃及伊蚊登革热病毒媒介能力的影响。一个表达芽孢杆菌来源的杀幼虫毒素的黄杆菌构建体将被评估其在天然树洞的实验室模型中的稳定整合和持久性，用qPCR和荧光报告器进行跟踪，并量化幼虫的死亡率。对蚊子致病性植菌株的持久性也将进行类似的研究。最后，我们将分析出蚊的稳定同位素富集特征，以追踪埃及伊蚊不同的幼虫栖息地，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.