Exploring New Reservoirs for Novel Bacteriophages with Therapeutic Potential Against ESKAPE Pathogens

A virus is an obligate, infectious nonliving particle of nucleic acid that replicates intracellularly via host DNA replication machinery, often to the detriment of the host. A bacteriophage is a type of virus that infects bacterial hosts, each having separate molecular tropism for a specific host. With rising antibiotic resistance in many medically-important bacterial pathogens, alternative methods for therapy will be essential for future treatment against these pathogens. One such therapy that holds promise is the use of bacteriophage. We seek to develop a high-throughput method of extracting bacteriophage from a given reservoir, including the environment or insects. Samples will be collected from tree holes around campus and subjected to filtration to remove “culturable” agents, which will serve as a viral source to infect specific bacterial hosts. In this study, we well use bacteria collectively known as ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) as they represent the leading global cause of hospital-acquired infections, and as many have developed antibiotic-resistance. The filtrate and host will be mixed in a molten top agar then poured onto a media plate to generate a bacterial lawn. If there are viruses with the specific tropism for the pathogen, after incubation the bacterial lawn will possess visible empty regions with dead cells known as plaques. Preliminary experiments demonstrate that E. coli and P. aeruginosa strain K have viruses that target and destroy the pathogens, as indicated by viral plaques. Isolation of these viruses via novel viral streak plating technique and molecular confirmation is currently underway. The search for more phage of other pathogens as well as the screening of the virome taken directly from mosquitoes collected from the environment is planned for later this spring and could reveal an untapped source of bacteriophage for potential phage therapy