(4mv) Nanocultures As an Assessment Tool for Microbial Dynamics

Background/Motivation: The term “viable but nonculturable bacterial cells (VBNC)", also known as “microbial dark matter (MDM),” is used to identify specific cells that exhibited metabolic activity and elongation upon nutrient administration yet were unable to form colonies on solid media. Failure to grow and study these VBNC microorganisms present in our diverse ecosystems poses a great challenge for researchers to understand their interactions and to discover novel penicillin-like compounds. Therefore, the need to access the MDM is imperative for the potential discovery of antibiotics, beneficial secondary metabolites, and naturally occurring therapeutic products. Methods: One approach to growing and retrieving environmental microbes is the miniaturization of culturing techniques which enables the growth and retrieval of environmental microbes by providing access to the microenvironment of microbial cells and reducing inter-species competition for space and resources. In the microBiointerface lab, we have developed a nanoculture system i.e., nanoliter-sized, water-oil-water W/O/W double emulsion, to isolate, confine, and observe synthetic microbial consortia. The nanocultures are formed by semi-permeable, highly robust, polymeric (PDMS-based) membranes. The semi-permeability allows the transportation of nutrients, waste produced by microbes, and other small molecules across the membrane. Furthermore, we can manipulate the polymer chemistry to create novel capabilities, including magnetization for successful retrieval of the nanocultures from the simulated microenvironment. Research Interests: Thus, our aim 1 focuses on designing a novel polymer mixture with a well-defined composition to generate a nanoculture system with desired pre-defined properties. To test the limits of our system, aim 2 will focus on growing multiple species and observing their interactions within the nanocultures, as well as examining their response to an external stimulus. It’s no surprise that the microbial communities are highly complex. Therefore, aim 3 will develop methods for cultivating microbes in situ using actinomyces as a model organism. The knowledge of microbial interactions within these communities can pave the way to develop targeted interventions and preventative treatments.