(811.6) Bacteriophage Isolation, Characterization and Bioinformatic Genetic Analyses against Actinomycetes Gordonia rubripertincta and Nocardia asteroides

Poster Board Number: A341

Jessica Stockert (Northern State University), Jon Mitchell (Northern State University)

Two of the biggest issues in the healthcare system today are antibiotic resistant bacteria and biofilms. Antibiotic-resistant bacterial strains are increasing, and current treatment options have become limited forcing medical facilities to search for alternatives. Recent studies utilizing phage therapy have provided promising results supporting the use of bacteriophages to treat bacterial infections. (Hatfull, 2021). A bacteriophage is a virus that only infects prokaryotic bacterial cells and therefore, potentially used to treat bacterial infections without risk of infecting the patient’s eukaryotic cells. This research focuses on the isolation and microbiological characterization of bacteriophages isolated from environmental samples against the Actinomycete host, Gordonia rubripertincta. Gordonia species are typically found in soil and aquatic habitats and have recently been identified as primary agents of infections in both humans and animals (Pope, et al., 2020).  Due to these organisms’ ability to form biofilms, infections have been found following various surgical procedures and applications of catheters (Arenskötter, et al., 2004).  Following a Gordonia-enrichment protocol, standard viral lab techniques such as serial dilutions, pick-a-plaque, and spot-plating were performed resulting in pure viral colonies. Genomic DNA was isolated and sequenced by the Pittsburgh Bacteriophage Institute. Genomes were annotated using Phamerator™, PECAAN™, and NCBI BLAST bioinformatics programs.  Host-range specificity studies and gene knock-out experiments were conducted using phylogenetically-related Actinomycetes to assist understanding of the molecular interactions between phage and host throughout the infection cycle.  Several bacteriophages were identified which exhibit both lytic and temperate life cycles against known clinical, biofilm-producing Actinomycete bacteria Nocardia asteroides and N. corallina. This research exemplifies the potential of using phage therapy to create an alternative treatment for antibiotic-resistant bacteria and biofilms within the healthcare system.