Staphylococcus aureus is a human commensal that can also cause systemic infections. This transition requires evasion of the immune response and the ability to exploit different niches within the host. However, the disease mechanisms and the dominant immune mediators against infection are poorly understood. Previously it has been shown that the infecting S. aureus population goes through a population bottleneck, from which very few bacteria escape to establish the abscesses that are characteristic of many infections. Here we examine the host factors underlying the population bottleneck and subsequent clonal expansion in S. aureus infection models, to identify underpinning principles of infection. The bottleneck is a common feature between models and is independent of S. aureus strain. Interestingly, the high doses of S. aureus required for the widely used “survival” model results in a reduced population bottleneck, suggesting that host defences have been simply overloaded. This brings into question the applicability of the survival model. Depletion of immune mediators revealed key breakpoints and the dynamics of systemic infection. Loss of macrophages, including the liver Kupffer cells, led to increased sensitivity to infection as expected but also loss of the population bottleneck and the spread to other organs still occurred. Conversely, neutrophil depletion led to greater susceptibility to disease but with a concomitant maintenance of the bottleneck and lack of systemic spread. We also used a novel microscopy approach to examine abscess architecture and distribution within organs. From these observations we developed a conceptual model for S. aureus disease from initial infection to mature abscess. This work highlights the need to understand the complexities of the infectious process to be able to assign functions for host and bacterial components, and why S. aureus disease requires a seemingly high infectious dose and how interventions such as a vaccine may be more rationally developed.

The rise of next generation sequencing is revealing a hidden diversity of temperate phages within the microbial community. While a handful of these phages have been well characterized, for the vast majority, the role of phage carriage, and especially multiple phage carriage, is poorly understood. The Liverpool epidemic strain of Pseudomonas aeruginosa is an aggressive pathogen in cystic fibrosis lung infections that has recently been found to contain several unique prophages within its genome. Here, we experimentally investigate the role of two of these phages in vivo, using an insect model of infection. We find that while no benefit is conferred by phage carriage in single bacterial infections, phages confer a large fitness advantage during mixed infections by mediating bacteria–bacteria competition. Differences between the two phages appeared to be associated with the rate at which the competitor acquired the phage, and therefore resistance. However, the advantage was greatest in the polylysogen, carrying both phages. These findings suggest that the LES phages may play an important role in host invasions and more generally show that the carriage of multiple phages may itself be beneficial by hindering the spread of resistance in rival bacterial populations.