New Approaches to Sponge-Microbial Symbioses

该提案旨在更好地了解海绵-细菌关联，特别是 确定涉及哪些细菌以及这些关联的稳定性。了解这样的 预计共生将产生具有药理活性的海绵或水产养殖的新方法 生产菌株的培养。这些方法将具有解决问题的巨大潜力 具有药用活性的海洋天然产物的供应问题。我们打算使用分子技术 例如变性梯度凝胶电泳 (DGGE) 来监测海绵中的微生物群落 以及应用最先进的化学分析以确定相应的次级 代谢物概况。此外，我们建议采用扩散生长的创新技术 Chambers（DGC）用于培养以前“无法培养”的海绵细菌。 DGC 本质上是 充满均质海绵组织和琼脂的半透膜。这可以固定细菌 聚合物膜内部，同时将细菌暴露于“海绵化学”。这些 技术将使我们能够检查：1）海绵次生代谢物的变化是否与 海绵相关微生物群落的变化； 2) DGC 允许培养以前“不可培养的” 海绵细菌； 3）共生海绵细菌或海绵本身产生新的、有趣的、 具有药理活性的化合物。除了关于海绵细菌共生的新见解（例如： 海绵细菌通才还是专家？），我们将创建一个潜在独特细菌菌株的库 使用新的 DGC 技术从海绵中分离出来。分离出来的海绵细菌将会生长出来， 提取，并筛选提取物中可能的新抗癌或抗感染代谢物。活动在任一 筛选之后将进行活性次生代谢物的分离和结构阐明。 为了有效治疗癌症或结核病（由多重耐药细菌引起）等疾病， 对开发新药很重要。尽管已经从海洋中分离出几种潜在的候选药物 有机体的进一步发展常常受到供应问题的阻碍。我们的研究旨在确定新的 从微生物来源寻找候选药物，同时开发新的培养技术来解决 供应问题

This proposal aims to gain a better understanding of sponge-bacterial associations and, in particular, to determine which bacteria are involved and how stable these associations are. Understanding such symbioses is predicted to yield new approaches for the aquaculture of pharmacologically-active sponges or the cultivation of the producing bacterial strains. Such methods would have enormous potential to solve the supply problem of pharmaceutically-active marine natural products. We intend to use molecular techniques such as Denaturing Gradient Gel Electrophoresis (DGGE) to monitor the microbial communities in sponges as well as to apply state-of-the-art chemical analysis in order to determine the corresponding secondary metabolite profiles. Furthermore, we propose to adopt the innovative technique of Diffusion Growth Chambers (DGCs) to the cultivation of previously "unculturable" sponge bacteria. DGCs are essentially semi-permeable membranes filled with homogenized sponge tissue and agar. This immobilizes bacteria inside the polymer membranes, while simultaneously exposing bacteria to the "sponge chemistry". These techniques will allow us to examine whether: 1) changes in sponge secondary metabolites correlate with changes in sponge-associated microbial communities; 2) DGCs allow culturing of previously "unculturable" sponge bacteria; and 3) symbiotic sponge bacteria or sponges themselves yield new, interesting, pharmacologically-active compounds. In addition to new insights on sponge-bacterial symbiosis (e.g. are ; sponge bacteria generalists or specialists?), we will create a library of potentially unique bacterial strains isolated from sponges with the new DGC technique. Isolated sponge bacteria will be grown out and extracted, and extracts screened for possible new anticancer or antiinfectant metabolites. Activity in either screen will be followed by the isolation and structure elucidation of the active secondary metabolites. To effectively treat diseases such as cancer or tuberculosis (caused by multi-drug-resistant bacteria), it is important to develop new drugs. Although several potential drug candidates have been isolated from marine organisms, further development is often hampered by a supply problem. Our research aims to identify new drug candidates from microbial sources and simultaneously develop new culturing techniques to address the supply problem