Abstract #M95

# M95
Bacteriophage identification from dairy environments.
R. H. Pittsley*1, T. Zubkov2, D. A. Sanders3, V. M. Blake1, K. G. Kornilow1, K. L. Adamson4, J. L. McKillip1, 1Department of Biology, Ball State University, Muncie, IN, 2Department of Chemistry, Ball State University, Muncie, IN, 3Markey Center for Structural Biology, Department of Biology, Purdue University, West Lafayette, IN, 4Elanco Animal Health, Clinton, IN.

Bacillus spp. are within a family of bacteria known to form spores for survival in extreme environments. The spores are resistant to temperature extremes, radiation, antibiotics, and many disinfectants. When ingested by or exposed to susceptible animal hosts, the spores can germinate and create biofilms or cause infections such as endophthalmitis, bacteremia, pneumonia, or gastroenteritis. The virulence potential makes these bacteria problematic in the food industry as pathogens and spoilers. As a protective measure, dairy farmers could utilize a known bacteriophage after pasteurization to ensure dairy product quality and safety. The goal of this research is to isolate and characterize unknown bacteriophage found in dairy milk. By determining host range, DNA sequence, and morphology, phage candidates will be identified that may have promise as agents for biological control of Bacillus spp. Raw milk, silage, tank water, manure, and soil samples were processed for phage extraction and enrichment. Results revealed that the phage isolated in this study demonstrated a broad host range across several common Bacillus spp., established that levels of CaCl2 were highly influential for phage uptake, indicated the presence of shared amplicons between the unknown phage and Bacillus phage deep blue (NC_031056) and Bacillus phage BCP8–2 (NC_027355), revealed that glycerol precipitation was more efficient for DNA sequencing, and unveiled that the isolated phage had an icosahedral head and presence of a sheath and tail fibers, measuring approximately 100nm together and sharing characteristics with the well-known T4 bacteriophage known to infect Bacillus spp. Further host range studies, genomic analyses, and PCR will reveal if this phage is novel and a possible future biological control agent, and/or whether the phage harbors antibiotic resistance genes that could have food safety and public health implications.

Key Words: bacteriophage, Bacillus spp., virulence