Phase variation of poly-N-acetylglucosamine expression in Staphylococcus aureus.

Polysaccharide intercellular adhesin (PIA), also known as poly-N-acetyl-ß-(1-6)-glucosamine (PIA/PNAG) is an important component of Staphylococcus aureus biofilms and also contributes to resistance to phagocytosis. The proteins IcaA, IcaD, IcaB, and IcaC are encoded within the intercellular adhesin (ica) operon and synthesize PIA/PNAG. We discovered a mechanism of phase variation in PIA/PNAG expression that appears to involve slipped-strand mispairing. The process is reversible and RecA-independent, and involves the expansion and contraction of a simple tetranucleotide tandem repeat within icaC. Inactivation of IcaC results in a PIA/PNAG-negative phenotype. A PIA/PNAG-hyperproducing strain gained a fitness advantage in vitro following the icaC mutation and loss of PIA/PNAG production. The mutation was also detected in two clinical isolates, suggesting that under certain conditions, loss of PIA/PNAG production may be advantageous during infection. There was also a survival advantage for an icaC-negative strain harboring intact icaADB genes relative to an isogenic icaADBC deletion mutant. Together, these results suggest that inactivation of icaC is a mode of phase variation for PIA/PNAG expression, that high-level production of PIA/PNAG carries a fitness cost, and that icaADB may contribute to bacterial fitness, by an unknown mechanism, in the absence of an intact icaC gene and PIA/PNAG production.

Staphylococcal biofilms: quest for the magic bullet.

The biofilm phenotype has been recognized only relatively recently in medical history but it has rapidly become clear that the development of many, if not the majority of bacterial infections depends upon the formation of a biofilm. Medical device-related infections are one of the clearest examples of biofilm-dependent infections. Bacteria proficiently adhere to and establish biofilms on synthetic surfaces, and to date, no material has proven to completely preclude bacterial adherence. Any inserted device can be colonized but intravenous catheters, due to their widespread use, are the most commonly colonized devices. As many as half a million catheter-related infections occur each year in the United States and the staphylococci, in particular, Staphylococcus aureus and Staphylococcus epidermidis, are the leading cause. Biofilms exhibit tolerance to biocides, chemotherapeutic agents, and host-immune defenses and subsequently, biofilm-associated infections are extremely difficult to treat, frequently chronic, and often recurrent, making them a confounding clinical problem. Development of an effective strategy for preventing and/or treating these infections is of paramount importance and consequently, the search for novel approaches to target the biofilm phenotype has exploded in recent years. Because the biofilm phenotype is complex, targets for antibiofilm approaches are numerous and this line of research is significantly expanding our knowledge about the biofilm mode of growth and its role in disease. This review highlights a number of antibiofilm approaches that are currently under investigation as novel interventions for staphylococcal infections.

Racioethnic diversity in the dynamics of the vaginal microbiome during pregnancy.

The microbiome of the female reproductive tract has implications for women's reproductive health. We examined the vaginal microbiome in two cohorts of women who experienced normal term births: a cross-sectionally sampled cohort of 613 pregnant and 1,969 non-pregnant women, focusing on 300 pregnant and 300 non-pregnant women of African, Hispanic or European ancestry case-matched for race, gestational age and household income; and a longitudinally sampled cohort of 90 pregnant women of African or non-African ancestry. In these women, the vaginal microbiome shifted during pregnancy toward Lactobacillus-dominated profiles at the expense of taxa often associated with vaginal dysbiosis. The shifts occurred early in pregnancy, followed predictable patterns, were associated with simplification of the metabolic capacity of the microbiome and were significant only in women of African or Hispanic ancestry. Both genomic and environmental factors are likely contributors to these trends, with socioeconomic status as a likely environmental influence.

The vaginal microbiome and preterm birth.

The incidence of preterm birth exceeds 10% worldwide. There are significant disparities in the frequency of preterm birth among populations within countries, and women of African ancestry disproportionately bear the burden of risk in the United States. In the present study, we report a community resource that includes 'omics' data from approximately 12,000 samples as part of the integrative Human Microbiome Project. Longitudinal analyses of 16S ribosomal RNA, metagenomic, metatranscriptomic and cytokine profiles from 45 preterm and 90 term birth controls identified harbingers of preterm birth in this cohort of women predominantly of African ancestry. Women who delivered preterm exhibited significantly lower vaginal levels of Lactobacillus crispatus and higher levels of BVAB1, Sneathia amnii, TM7-H1, a group of Prevotella species and nine additional taxa. The first representative genomes of BVAB1 and TM7-H1 are described. Preterm-birth-associated taxa were correlated with proinflammatory cytokines in vaginal fluid. These findings highlight new opportunities for assessment of the risk of preterm birth.

Regulation of the intercellular adhesin locus regulator (icaR) by SarA, sigmaB, and IcaR in Staphylococcus aureus.

The staphylococcal accessory regulator SarA and the alternative sigma factor sigma(B) have been previously identified as positive regulators, and IcaR as a negative regulator, of icaADBC expression. Here, we show that in Staphylococcus aureus SarA and sigma(B) are also required for icaR expression and that IcaR does not have a significant effect on its own expression.