LARVAL MOSQUITO FEEDING AND MICROBIAL INTERACTIONS

蚊子是最重要的媒介昆虫， 它们传播的不同病原体的数量， 这些疾病造成的健康问题的严重性。 由于 蚊子传播的疾病在世界范围内的复苏， 基于正确的生态学理解的蚊子控制 蚊子生物学是必要的。 蚊子幼虫呈现出一种诱人的 这些方法的目标。 幼虫期微生物群落的性质 蚊子的栖息地是了解蚊子 幼虫利用天然食物，生长并发育成成虫。 很长 我们所提出的研究的长期目标是了解 微生物作为食品和潜在的控制剂， 蚊子幼虫 我们提出的研究的一个隐含目标是 扩大蚊子幼虫中细菌群落的特征 栖息地 蓝藻杀幼虫毒素基因克隆的研究进展 和其他主持人建议，这种方法可以提供一种方法， 持续的幼虫控制 我们已经证实革兰氏阴性杆菌 在幼虫的栖息地发现了种类繁多的细菌。 广泛的主机- 范围质粒现在可用于将DNA转移到革兰氏阴性菌中， 细菌 因此，另一个研究目标是从细菌中选择细菌。 幼虫栖息地作为DNA转移和基因重组候选物 苏云金芽孢杆菌血清型cryIVD基因的表达 Israelensis（编码72 kDa杀幼虫毒素）。 我们将初步 将质粒携带的基因插入到实验室菌株恶臭假单胞菌中，和 然后将这一努力扩大到包括选定的环境革兰氏阴性菌， 我们已经分离并鉴定了 蚊子栖息地 最后，我们的目标是将重组体引入 实验室微观栖息地并检查这些栖息地的持续性 利用耐药性标记和基于PCR的 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.