05. Novel Technique for the Isolation of Bacteriophage from Environmental Samples

Abstract

Bacteriophage (phage) are viruses that infect bacteria and are proposed to be the most copious organism on the planet. Study of phage leads to insights into novel therapeutics of antibiotic-resistant bacterial infections, as well as the development of molecular tools such as the CRISPR/Cas9 system. In order to further knowledge of phage biology, it is beneficial to isolate, identify, and study novel phage and their properties. Current phage isolation strategies rely on prolonged processing and luck, identifying single isolates at a time. Therefore, the establishment of a protocol with a higher throughput will enable scientists to isolate more phage in a shorter timeframe as it enables scanning more samples simultaneously, reducing time and increasing the probability of successful phage isolation. It is known that high bacterial cell density correlates to living cells and low density equates to dying cells. The presence of phage is identified by bacterial cell death. Therefore, it is our hypothesis that bacterial culture density decreases can be used to detect the presence of phage in multiple samples simultaneously, allowing for the efficient identification of phage for further study. Previous work has demonstrated that the changes in density can be measured using standard photometric assays and equipment. These measurements display remarkable sensitivity at low concentrations of phage. Future studies will analyze the sensitivity of this assay and its applications for novel phage isolation. This methodological approach described and developed here will be shared with the scientific community for improvement of phage isolation and study.