New Approaches to Sponge-Microbial Symbioses

该提案旨在更好地了解海绵细菌协会，特别是， 确定哪些细菌参与以及这些关联的稳定性。理解这些 共生预计将产生新的方法，用于水产养殖的药理活性海绵或 生产菌株的培养。这些方法将具有巨大的潜力来解决 医药活性海洋天然产品供应问题。我们打算用分子技术 例如变性梯度凝胶电泳（DGGE），以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 代谢产物谱。此外，我们建议采用扩散生长的创新技术， 培养室（DGC）培养以前“不可培养”的海绵细菌。DGC基本上是 填充有均质化海绵组织和琼脂的半透膜。这样可以固定细菌 在聚合物膜内，同时将细菌暴露于“海绵化学”。这些 技术将使我们能够检查是否：1）海绵次生代谢物的变化与 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;和3）共生海绵细菌或海绵本身产生新的，有趣的， 药理活性化合物。除了对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的独特细菌菌株库， 用新的DGC技术从海绵中分离出来。分离的海绵细菌将生长出来， 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 筛选后将进行活性次级代谢产物的分离和结构解析。 为了有效治疗癌症或结核病（由多重耐药细菌引起）等疾病， 重要的是开发新药。虽然已经从海洋中分离出了几种潜在的药物候选物， 生物体的进一步发展往往受到供应问题的阻碍。我们的研究旨在发现新的 同时开发新的培养技术，以解决 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