Isolation and characterisation of phages infecting gram positive food bacteria

Bacteriophage (phage), virus of bacteria, has been proposed as a mean to inactivate bacteria that are pathogens of humans. Applied prophylatically to food, phage might decrease the numbers of potential pathogens we ingest. Much active research on using the phages of bacteria to control Gram negative foodborne pathogens are described in the literatures, but comparatively little research describes the phages of Gram positive bacteria and their use as biocontrol agents on food. In this work, previous undescribed phages, able to infect Bacillus cereus and Listeria monocytogenes, were isolated from soil and ruminants faecal material, respectively. As the first step in assessing their potential as biocontrol agents, the isolated phages were purified, concentrated and characterised (albeit to different degrees). The Bacillus phages had a narrow host range while the Listeria phages had a broad host range. Listeria phages also infected L. monocytogenes 2000/47, a strain which recurs in New Zealand clinical cases. Both Bacillus and Listeria phages appeared to be of the Myoviridae family judging by their structure in electron micrographs. The Bacillus FWLBc1 and FWLBc2 phages were lytic phages with a latent period of 106 and 102 min at 37°C, and an average burst size of 322 and 300 phages per infected cell, respectively. Moreover, they both had genomes of approximately 134 kb. All newly isolated and characterized phages were chloroform resistant and survived storage better at 4°C than at room or freezing temperatures. Bacillus phages significantly reduced the bacterial population in mashed potatoes within 24 h at room temperature, when applied at a phage to host ratio of 1000. Listeria phages rapidly inactivated the host population to a low optical density. The findings of this thesis will add to the current knowledge of phages in the context of various environmental conditions for different bacteria and will demonstrate the potential of phages as food safety biocontrol agents