Bacteriophage modulation of mosquito microbiota

摘要 蚊子相关细菌对幼虫的发育、寿命、媒介能力有重大影响 以及交配偏好。多项研究已经破译了相关细菌的分类多样性 不同种类的蚊子。已有研究表明，微生物区系的改变会影响蚊子的多种生活特征。 尽管有这些研究，单个细菌分类群的功能作用以及它们之间的相互作用 蚊子的肠道仍不清楚。为了研究来自复杂群落的单个细菌的作用， 需要对蚊子微生物区系进行有针对性的调节。噬菌体提供了一个完美的工具 从微生物群中选择性地去除目标细菌。这项提案旨在利用噬菌体来 选择性地调节蚊子的微生物区系，研究其对蚊子各种生活史特征的影响。 根据文献和我们自己的研究，我们已经确定了五个经常联系在一起的细菌分类群 不同种类的蚊子。在目标1中，我们将从各种环境中分离出噬菌体 能够感染选定蚊子肠道细菌的来源。在我们的初步实验中，我们已经 成功分离出多种能够选择性感染蚊子肠道细菌的噬菌体。 在目标2中，我们将使用分离的噬菌体选择性地调节成年伊蚊和 按蚊。这种改变的微生物区系对蚊子寿命、繁殖力和媒介的影响 将对能力进行研究。在目标3中，我们将使用分离的噬菌体选择性地调节 幼虫繁育水中的微生物区系。我们将评估改变的微生物区系对幼虫发育的影响， 以及成虫的媒介能力、寿命和繁殖力。我们关于灵知生菌的初步数据 幼虫已经表明，在幼虫繁殖水中添加噬菌体会影响幼虫的发育。 该项目将是首次在蚊子微生物区系研究中使用噬菌体，并具有 有可能发展成一种控制蚊媒疾病的新技术。

SUMMARY Mosquito associated bacteria have a major impact on larval development, life span, vector competence and mating preferences. Multiple studies have deciphered the taxonomic diversity of bacteria associated with various mosquito species. It has been shown that altered microbiota affects multiple mosquito life traits. Despite these studies, the functional role of individual bacterial taxa and their interactions with each other in the mosquito gut remains unknown. To study the role of individual bacteria from a complex community, targeted modulation of the mosquito microbiota is desired. Bacteriophages provide a perfect tool for selective removal of targeted bacteria from microbiota. This proposal aims to utilize bacteriophages to selectively modulate the mosquito microbiota and study its effect of various mosquito life-history traits. Based on literature and our own studies, we have identified five bacterial taxa that are frequently associated with various mosquito species. In aim 1, we will isolate bacteriophages from various environmental sources capable of infecting selected mosquito gut bacteria. In our preliminary experiments we have already successfully isolated multiple bacteriophages capable of selectively infecting various mosquito gut bacteria. In aim 2, we will use isolated bacteriophages to selectively modulate the microbiota of adult Aedes and Anopheles mosquitoes. The effect of this altered microbiota on mosquito life span, fecundity and vector competence will be studied. In aim 3, we will use isolated bacteriophages to selectively modulate the microbiota in larval breeding water. We will evaluate the effect of altered microbiota on larval development, and vector competence, lifespan and fecundity of adult mosquitoes. Our preliminary data with gnotobiotic larvae has shown that addition of bacteriophages to larval breeding water can affect the larval development. This project will be the first ever use of bacteriophages in the mosquito microbiota research and has the potential to be developed into a novel technique to control mosquito borne diseases.