Larval mosquito feeding and microbial interactions

来自水生、异养、幼虫栖息地的成蚊的生长和繁殖是主要的 成年人的存在和密度的决定因素，并强烈影响矢量能力，但它受到限制 将无菌有机材料转化为可摄取和可消化的微生物生物质。树洞 几种蚊子的栖息地是我们研究这一现象的模型系统。我们的长期目标是 量化蚊子繁殖的生物学基础和限制，确定 利用这些资源，详细说明幼虫生长的现实模型，并进行遗传转化 幼虫栖息地的细菌产生抗幼虫毒素。3个具体目标的研究方法包括 一个功能性纯性（无菌）实验系统，其中个别食物来源，包括 可以提供微生物;利用稳定同位素分析（13 C/12 C; 15 N/14 N）来跟踪 幼虫的食物来源;以及微生物群落的分子表征和操纵。 微生物代谢的生长促进作用和腐殖物质的抑制作用，将是 分析了提出了反应规范的概念，其中发育时间和变态质量 （化蛹）被假定为灵活的生活史性状营养限制到化蛹窗口。一 本文详细阐述了生长模型，其中阐述了食物转化效率，因此我们将 反复测试对各种质量的食物来源的生长反应。四种蚊类共用 相同的幼虫栖息地和资源（三列黄蚊Ochlerotatus triseriatus，黄蚊Ochlerotatus candidus，白纹伊蚊Aedes albopictus，和白纹伊蚊Aedes tiseriatus）， 将比较尖音库蚊（所有人类病原体的载体），以评估在以下条件下的生长率： 激烈的种内和种间竞争，以预测竞争结果和潜力 生态置换，并量化幼虫食物利用的相对效率。 微生物群落（细菌、真菌和原生动物）将通过微生物生物量进行评估， 生长测量，功能基因阵列的使用，以及聚合物降解酶的估计， 包括木聚糖酶、纤维素酶、几丁质酶。微生物的黄杆菌属组分的操作 将进行与分子遗传工具的共同体，以表达感兴趣的蛋白质和基因， 包括微生物毒素基因和协同几丁质酶。

The growth and production of adult mosquitoes from aquatic, heterotrophic, larval habitats is the primary determinant of adult presence and density, and strongly influences vectorial capacity, but it is constrained by conversion of refractile organic material into ingestible and digestible microbial biomass. The tree hole habitat of several mosquito species is our model system to study this phenomenon. Our long term goals are to quantify the biological basis for and constraints on mosquito production, to identify the efficiency of utilization of these resources, to elaborate a realistic model of larval growth, and to genetically transform bacteria in larval habitats to produce anti-larval toxins. Research approaches across 3 specific aims include a functionally axenic (germ-free) experimental system into which individual food sources including microorganisms can be provisioned; the utilization of stable isotope analyses (13C/12C; 15N/14N) to track larval foods to their sources; and molecular characterization and manipulation of the microbial community. Growth enhancing effects of microbial metabolism, and inhibiting effects of humic substances, will be analyzed. A reaction norm concept is proposed in which developmental time to and mass at metamorphosis (pupation) are postulated to be flexible life history traits nutritionally constrained into a pupation window. A growth model is elaborated herein, in which food conversion efficiencies are elaborated, thus we will iteratively test growth responses to food sources of various qualities. Four mosquitoes species sharing the same larval habitat and resources (Ochlerotatus triseriatus, Ochlerotatus japonicus, Aedes albopictus, and t Culex pipiens, all vectors of human pathogens) will be compared to assess growth rates under conditions of intense intra- and interspecific competition, both to make predictions of competitive outcomes and potential for ecological displacement, and to quantify comparative efficiency of utilization of larval foods.The role of the microbial community (bacteria, fungi, and protozoans) will be assessed through microbial biomass and growth measures, the use of functional gene arrays, and with estimates of polymer degradation enzymes, including xylanases, cellulases, chitinases. Manipulation of the Flavobacterium component of the microbial community with molecular genetic tools to express proteins and genes of interest will be conducted, including microbial toxin genes and synergistic chitinases.