Investigation of the effect of oxygen and temperature levels on E.coli K1 phages infection

"The problem of antimicrobial resistance is major and the need to identify alternative methods to antibiotics is obvious. One of the potential alternatives is phage therapy, the use of bacteriophages with high specificity against the targeted pathogen. The interest in phage therapy has grown lately and therefore different factors that can increase or decrease the phages' killing efficiency, are becoming interesting to study. In this project, we aim to investigate the effect of different parameters, such as oxygen levels and temperature, to the killing efficiency and infection ability of bacteriophages. It has been shown that both oxygen and temperature levels contribute to the infection ability of bacteriophages. To further investigate what happens inside the human body, where the conditions are fluctuating, we will use a model system previously established in Sagona lab, consisting of E.coli bacteriophages, human cells and E.coli strains.
The specific objectives are:
1. Examine using microbiology and phage biology methods the killing efficiency of bacteriophage in oxygen rich and oxygen-limited environments and high and low temperatures.
2. Investigate the effect of the above conditions in the health of human cells, using specific assays for cytotoxicity.
3. Examine the interaction between phage, its host and human cells, under different conditions.
4. Develop methodology to determine the scale and scope of innate immune responses in human cells under the different conditions against the host and phage.
5. Compare the different parameters by statistical analysis to conclude in the optimal conditions, where the phage presents the highest killing efficiency along with the least innate immune response.
A future scope of this project is to establish a system and gain knowledge on this, so that the same approach can be implemented broader. A combination of molecular biology, microbiology, immunology and human cell culture methods will be used."