LARVAL MOSQUITO FEEDING AND MICROBIAL INTERACTIONS

蚊子是最重要的媒介昆虫群，由于 它们传播的不同病原体的数量和 这些疾病造成的健康问题的严重性。由于 全球蚊媒疾病死灰复燃，新方法 基于对蚊子的良好生态理解的蚊子控制 蚊子生物学是必要的。幼虫蚊子呈现出诱人的 这类方法的目标。幼虫体内微生物群落的性质 蚊子栖息地是了解蚊子如何 幼虫利用天然食物，成长并发育成成虫。一个很长的 我们提议的研究的术语目标是了解 微生物作为食品和潜在的控制剂发挥作用 蚊子幼虫。我们提出的研究的一个隐含的目的是 扩展幼虫蚊子中细菌群落的特征 栖息地。蓝藻杀幼虫毒素基因克隆成功 和其他宿主建议，这种方法可以提供一种方法 持续的幼虫控制。我们已经记录了革兰氏阴性 在幼虫的栖息地中发现了不同种类的细菌。广泛的主机- Range质粒现在可用于将DNA转移到革兰氏阴性菌 细菌。因此，另一个研究目标是从 作为DNA转移和基因重组候选者的幼虫栖息地 苏云金芽孢杆菌血清型CryIVD基因的表达 以色列(编码一种72 kDa的杀幼虫毒素)。我们最初会 将质粒携带的基因插入恶臭假单胞菌实验室菌株，并 然后将这一努力扩大到包括选定的环境革兰氏阴性 我们已经分离和鉴定的细菌污渍 蚊子栖息地。最后，我们的目标是将重组子引入 实验室微观世界栖息地，并检查这些栖息地的持久性 利用耐药标记物和聚合酶链式反应 CryD基因片段的检测。

Mosquitoes stand out as the most important group of vector insects due to the number of different disease agents they transmit and the magnitude of health problems these diseases cause. Owing to the resurgence of mosquito-borne disease worldwide, new methods for mosquito control based upon sound ecological understanding of mosquito biology is needed. Larval mosquitoes present a tantalizing target for such methods. The nature of microbial communities in larval mosquito habitats is a prime element in understanding how mosquito larvae exploit natural food, grow, and develop into adults. A long term objective of our proposed research is to understand the role that microorganisms play as food items and potential control agents for mosquito larvae. An implicit aim of our proposed research is to expand characterization of the bacterial community in larval mosquito habitats. Successes in cloning larvicidal toxin genes in cyanobacteria and other hosts suggest that this method may provide a method of sustained larval control. We have documented that gram negative bacteria of diverse general are found in larval habitats. Broad host- range plasmids are now available for DNA transfer into gram-negative bacteria. Thus, another research objective is to select bacteria from larval habitats as recombinant candidates for DNA transfer and gene expression of the cryIVD gene of Bacillus thuringiensis serovar. Israelensis (coding for a 72kDa larvicidal toxin). We will initially insert the plasmid-borne gene into a lab-strain pseudomonas putida, and then expand this effort to include selected environmental gram-negative bacterial stains which we have already isolated and characterized from mosquito habitats. Finally, we aim to introduce recombinants into laboratory microcosm habitats and examine the persistence of these organisms by utilizing drug resistance markers and PCR-based detection of the cryD gene fragment.