Culture Dependent Analysis of Microbial Communities from Raspberry Rising Cave Soil Samples

The aim of this study was to elucidate the microbial diversity of soil samples collected from the Raspberry Rising Cave area implementing cultivation dependent approaches. The Raspberry Rising Cave is a marble cave system located in the Glacier National Park of Canada in the Mount Tupper area. In this study, bacterial colonies were isolated from seven different sampling areas from the Raspberry Rising Cave. Bacterial colonies were isolated by taking each soil sample and spinning them in volumetric flasks overnight at 8°C on a shaking incubator at 150 RPM covered by sterile deionized water. The soil samples supernatant was serially diluted and then plated on three different types of media, R2A, Hickey-Tresner, and Actinomycetes the following day using a glass bead spread plate method. These spread plates were incubated at 8°C and underwent sub-culturing to obtain pure bacterial isolates. As a result, a total of 103 bacterial isolates were grown at 8°C for approximately 1-2 weeks. These bacterial cultures were then grown in liquid broth at 8°C for seven days on a shaking incubator at 150 RPM and subjected to DNA extraction, which were resolved on 0.8% ethidium bromide-agarose gels. The amplification of the DNA products from those 103 bacterial samples is currently being performed by polymerase chain reaction (PCR), with subsequent 16S rRNA sequencing to take place to identify the isolates that were cultivated. Antimicrobial screening will also occur against multidrug resistant bacterial strains to identify any antimicrobial activity the bacterial isolates may possess