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Table of contents
Chapter 1. Bibliographic study
1.1 Salmonella Enteritidis in eggs, a matter of public health
1.2 The powerful antibacterial defence mechanisms of egg white
1.3 Iron in the fight between Salmonella Enteritidis and egg white
1.4 Approaches set in the PhD proposal
Chapter 2. Material and methods
2.1 Reagents and chemicals
2.2 In vitro DNA procedures
2.3 Bacterial transformation
2.4 Protein procedures
2.5 Production of the human lipocalin and its homologues found in egg white
2.6 Measure of biomolecular interactions between lipocalins and siderophores
2.7 Genetic engineering techniques
2.8 Siderophore detection assay
2.9 Measurement of Salmonella Enteritidis growth dynamics
2.10 Statistical analysis
Chapter 3. Isolation of recombinant egg-white lipocalins
3.1 Bioinformatic analysis of lipocalin-2 homologues found in egg white
3.2 Overproduction of the lipocalin-2 and egg white lipocalins
3.3 Purification and characterisation of the overproduced lipocalins
3.4 Conclusion and future work
Chapter 4. Quantification of egg-white lipocalins and determination of their siderophore-binding activity
4.1 Egg white contains micro-Molar levels of all three lipocalins
4.2 Ex-FABP binds enterobactin with high affinity and strong preference for the ferrated form
4.3 Conclusion and future work
Chapter 5. Determination of the role of egg-white lipocalins in inhibiting siderophore-dependent iron sequestration by Salmonella Enteritidis
5.1 Library construction of mutants deficient in iron acquisition systems
5.2 Provision of Ex-FABP inhibits growth of a salmochelin-deficient Salmonella Enteritidis mutant in standard growth media
5.3 The ability to synthesise siderophore does not support Salmonella Enteritidis persistence in egg-white media
5.4 Ex-FABP antibacterial activity (via its siderophore-binding capacity) observed in standard growth media is not observed in egg-white media
5.5 Conclusion and future work
Chapter 6. General discussion
6.1 Salmonella Enteritidis metal acquisition in egg white
6.2 Could Ex-FABP be a component of the hen immune defence?
6.3 Conclusion and future work
Conclusion
References
