This process is based upon numerous features of the bacterial cell including alterations in their metabolism and physiology, the presence and nature of surface structures, and the general physical properties of the bacterial cell. The process of biofilm formation is defined in stages and each of these has

specific features and profiles [2]. Put simply, under stressed conditions bacterial cells can switch from a free-living and a rapidly dividing phenotype to an altered metabolic NVP-BGJ398 mw form associated with cell-cell aggregation and attachment to a surface. There are then early, mid, and late stages for the maturation of a bacterial biofilm. The particular stresses that induce a change in lifestyle and subsequently the process of biofilm formation are poorly

defined for many pathogenic bacteria, however antibiotic usage is certainly one, nutrient starvation and oxidative stress are others [4]. These conditions or signals do seem to be specific for different species. Despite some previous disagreement about the ability of H. influenzae to form a biofilm [6], there is now overwhelming evidence that H. influenzae use biofilm formation for survival within the host and certainly in their colonization of the host [7–13]. There are elements of H. influenzae which seem to be induced and therefore important for biofilm formation [13]. There are numerous examples of studies that have shown that iron uptake is central to growth within a biofilm [14–20]. There is a need to further characterise the differences between biofilm-forming and non-biofilm-forming see more check details isolates of H. influenzae. This can be accomplished through a comparison of the genetic and transcriptomic differences between H. influenzae strains

that respond to stresses by forming a biofilm, and those that continue to grow under those conditions without forming a biofilm. Changes in pH provides SPTLC1 a suitable stressor, being central to its colonisation of different anatomical niches, and identification of the molecular pathways that vary between such isolates would be significant in our understanding of H. influenzae pathogenesis. H. influenzae strains and isolates display more variation than many other pathogens and underpinning the basis for the strain-specific actors that underlie their biofilm formation (recently reviewed [21, 22]). Indeed, coupled to this, there are many features of the H. influenzae physiology [23–25] and stress response [26–30] that indicate that this particular host-adapted bacterium has unique molecular mechanisms for survival in the various locations of its host that it can exist. The pH is known to be elevated in the middle ear, compared to other parts of the body [31, 32] and in this niche there is some evidence that it is pH that induces particular isolates of H. influenzae to form a biofilm [33]. We have assessed the response of different clinical isolates of H.