Understanding the formation of Pseudomonas aeruginosa biofilms in urinary catheters (360G-Wellcome-211751_Z_18_Z)

This proposal uses advanced microscopy and molecular techniques to examine the role of alginate in the attachment of Pseudomonas aeruginosa, and subsequent biofilm formation, on urinary catheters. There is an urgent clinical need for improved catheter management and the development of anti-biofilm materials, however approaches to date have failed and a lack of understanding of biofilm development may contribute to this. In this work, we intend using a simple laboratory model system to generate biofilms on urinary catheters, which can be tracked directly over time using episcopic differential interference contrast microscopy. Transposon mutants (obtained from the PA Two Allele Library) will be used and compared to the type strain, PAO1. In addition, Gibson Assembly will be used to generate knockout mutants for the same genes. The biofilm forming capability of each mutant and type (transposon vs knockout) will be assessed and compared. Transposon mutants have the potential to still produce peptides, with unknown form and function. Such peptides could affect biofilm development. In contrast, knockout mutants remove the target gene sequence. This project will provide information on general biofilm development, the role of alginate, and also indicate whether knockout mutants should be used for understanding the role of specific genes.