To evaluate extracts for anticancer and anti-tuberculosis activities. Extracts from all individual microorganisms will be obtained and evaluated in a series of anticancer and anti-tuberculosis assays.

To determine the presence (or absence) of known metabolites in active extracts. All active extracts will be fractionated and analyzed by LC MS to identify any known metabolites.

Methodology
To isolate and grow previously unculturable aquatic microorganisms. Microbial Isolated: we will collect sediments and water from various Lake Michigan habitats and use them to establish a simulated natural environment in aquaria. We will then use this environment to isolate “unculturable” microorganisms using diffusion chambers. Purified isolates will be identified by 16S rRNA sequencing. Identified isolates will be re-grown to obtain a sufficient amount of biomass for biological and chemical evaluation. We intend to evaluate a phenomenon called “domestication” to enable us to grow these microorganisms on standard Petri dishes.

Rationale
The proposed research will explore and take advantage of a vast, but untapped resource existing in Lake Michigan – previously unculturable microorganisms. The majority of microorganisms from all environments resist cultivation in the laboratory. We can now, however, access this microbial diversity by employing a recently developed method: diffusion chambers in simulated natural environments. The study of these organisms will enhance our understanding of the diversity aquatic microorganisms in Lake Michigan. We will also be able to investigate the chemical and biochemical potential of these previously uncultured aquatic microorganisms, which we believe will validate our long range hypothesis that by accessing these unculturable microorganisms we will be able to gain access to an enormous source of novel bioactive organic molecules.