Development and Application of Strategies to Generate Bacteriophage Resistant Strains for Use in Milk Fermentation Processes

Abstract

The objectives of this project were firstly, the identification of natural phage resistance systems for exploitation, secondly, the development of methodologies to utilise these systems to improve the bacteriophage resistance of starter strains for use in milk fermentation processes, and thirdly, the actual application of these methodologies to improving starter strains.

The main conclusions were as follows:

Three new natural plasmid (DNA)-associated bacteriophage resistance systems were identified at Moorepark. The detailed genetic makeup of the phage resistance plasmid (pMRC01) was elucidated. Bacteriophages currently evolving in the industrial cheese-making environment were monitored to facilitate the judicious choice of phage resistance systems for use in commercial starter cultures which can more effectively target the documented problematic phage types. Two highly virulent phages targeting important cheese starters were identified in the industrial cheese-making environment. A reliable food-grade method to facilitate the transfer of phage resistance systems to cheese-making starter strains was developed. This is based on bacteriocin immunity-linked phage resistance. Phage resistant cheese starter cultures were developed through natural selection and by molecular manipulation using phage resistance plasmids. The phage resistance plasmid pMRC01 was introduced to 31 cheese starter strains.