Life science research will on focus on the identification and characterization of novel organisms living deep underground. The emphasis will be on discovery of genes that have applied applications in industry or medicine. Organisms living in extreme environments, such as deep underground, have the greatest potential for the development of intellectual property leading to commercialization. A molecular approach to these studies may lead to the discovery of novel antibiotics and high-quality lignocellulolytic enzymes that will lead to commercialization. The research focus leverages the research capacity at BHSU in genomics and collaborations across the state.

Antimicrobial drug resistance in bacterial and fungal pathogens is currently a major problem in medicine. The Center for Disease Control estimates that over 2,000,000 hospitalizations and 23,000 deaths result from infections by antibiotic-resistant microbes. Despite the critical need for new groups of antibiotics, the rate of discovery of novel antimicrobial drugs has declined considerably over the past 30 years. One source of antimicrobial drugs is antimicrobial producing microbes. The majority of antimicrobial producing microbes have yet to be isolated, in large part due to difficulties in isolating them in pure culture [1]. The Actinobacteria group of microbes produce the majority of known antimicrobial drugs. Biofilms located at the 17 Ledge region of the 4850 level in Sanford Lab have been shown to be rich in Actinobacteria [2].

One way to increase the diversity of microbes which can be isolated and cultured is to use in-situ culturing techniques, in which a sample containing microbes and suitable media is enclosed in a small diffusion chamber placed within the microbe’s natural habitat [3]. Gavrish et al. developed a unique in-situ diffusion chamber, which selectively isolated novel Actinobacteria, which produce the majority of known antimicrobial drugs, from soils [4].

We propose the use of in-situ diffusion chambers placed in two habitats in the lower levels (4100 to 4850 ft) of the Sanford Lab as a way to isolate, bring into cultivation, and identify groups of Actinobacteria which may produce novel antimicrobial compounds, and perform initial screening of these compounds against target bacteria, including Staphylococcus aureus and Escherichia coli.

To perform initial screening of the isolates for antibacterial activity, isolates will be grown in several different kinds of agar media and the cultures will be dried down, possible antimicrobial compounds extracted with DMSO, and the extracts recovered by filtration in a centrifuge into a 96-well plate [5]. DNA will be extracted from the isolates and DNA will be sequenced to determine the likely species or strains represented in the samples. Each extract will also be applied to a petri plate pre-inoculated with the target bacterium (Staphylococcus aureus or Escherichia coli) and incubated 12-48 hrs at 37o C. Zones of growth inhibition surrounding the points of application of the extract indicates that the isolate produces antimicrobial compounds.