The second group received the complemented strain, HI2210, along with HI2206 (Figure 4C). The last group of animals was infected with R2866 and HI2210 (Figure 4D). The hfq mutant exhibited significantly lower bacteremic titers throughout the course of the experiment when compared to either the wild type or the complemented mutant strains. As shown by the competitive index, the ∆hfq strain was approximately a 100-fold lower than the wild type strain by day one and all animals had completely PS-341 price cleared the mutant strain by day 3 post infection. Similar differences were observed in the animals infected with the ∆hfq complement strain and the ∆hfq strain, indicating the complement
strain exhibits a reversal of 3-MA clinical trial the mutant phenotype, however, there was not a complete reversal of the mutant phenotype (Figure 4D). The wild type strain did significantly out compete the complemented strain on days 2 and 3 post-infection. Complementation only partially restores the in vitro growth phenotype, and since the in vivo environment is likely to be more rigorously restricted for essential nutrients, the difference between wild
type and complemented strain may be exacerbated in vivo. The role of Hfq during infections of H. influenzae is not clear. In other organisms several sRNAs that interact with Hfq have been shown to be important in the regulation of genes involved in pathogenesis . It is currently unknown if H. influenzae has sRNAs that are important in pathogenesis. However, our animal studies suggest that in the absence of Hfq, certain genes important in establishing infection are likely affected.
Presumably, these genes are regulated by sRNAs either directly or indirectly and require Hfq to function properly. However, during the virulence studies there was no observed difference in either animal model, indicating that the ∆hfq mutant was able to grow within the host Amino acid environment. The defect is apparently limited to the occupation of specific niches within the host that are unavailable in a mixed infection due to the presence of the wild type strain. The loss of post-transcriptional regulation in the ∆hfq mutant leads to the inability of the bacteria to adapt to the host environment and compete successfully for the specific niches that are required for pathogenesis. The observations made in this study indicate there is a decrease in fitness in the animal models, and this phenotype is conserved across different strains. This effect may be partially explained by the selleck chemical impact of hfq mutation on acquisition of essential nutrients such as heme. While we did not address biofilm formation in the chinchilla middle ear, the possibility remains that mutation of hfq may influence adherence/biofilm formation in the microenvironment. A better understanding of the nutrients available in the host is necessary for a comprehensive explanation of the decrease in fitness identified in the mutant strain.