Objective To investigate the influence of menses on the vaginal microbiota and determine whether tampons that differ in material composition influence these bacterial communities in different ways.Design A single-centre trial with randomised, complete block design. Setting Procter & Gamble facility.Sample Seven self-declared healthy, female volunteers of reproductive age.Methods Volunteers used a pad and two types of tampons during the study, one product exclusively each month for three sequential menstrual cycles. During menses and once each midcycle, vaginal bacterial community composition was characterised by cultivation-independent methods based on pyrosequencing of V1-V2 variable regions of 16S ribosomal RNA genes.Main outcome measures Changes in the species composition, abundance and diversity in vaginal bacterial communities over time and between treatments.Results The vaginal microbiotas of all seven women were dominated by Lactobacillus spp. at mid-cycle, and the compositions of those communities were largely consistent between cycles. Community dynamic patterns during menses varied considerably and were more or less individualised. In three of the seven women the community diversity during pad use was significantly different from at least one tampon cycle.Conclusions Changes in the composition of the vaginal microbiota during menses were common, but the magnitude of change varied between women. Despite these changes, most communities were capable of resuming a composition similar to previous midcycle sampling times following menstruation. Overall we conclude that the two tampons tested do not significantly impact the vaginal microbiota in different ways; however, larger studies should be performed to confirm these findings.

Prior studies suggest Staphylococcus aureus exotoxins are not produced when the organism is cultured in human blood. Human blood was fractionated into plasma and water-lysed red blood cells and demonstrated that mixtures of α and β globins of hemoglobin (as low as 1 ug/ml) inhibited S. aureus exotoxin production while increasing production of protein A and not affecting bacterial growth. Pepsin but not trypsin digestion destroyed the ability of α and β globin to inhibit exotoxin production. Exotoxin production by both methicillin-resistant and susceptible organisms was inhibited. Production of streptococcal pyrogenic exotoxin A by Streptococcus pyogenes was unaffected by α and β globin chains, but was inhibited when produced in S. aureus. Use of isogenic S. aureus strains suggested the targets of α and β globin chains, leading to inhibition of staphylococcal exotoxins, included the two component system SrrA-SrrB. Delta hemolysin production was also inhibited, suggesting the two component (and quorum sensing) system AgrA-AgrC was targeted. The α and β globin chains represent promising molecules to interfere with the pathogenesis of serious staphylococcal diseases.Staphylococcus aureus causes large numbers of human diseases, primarily initiated by colonization of mucosal surfaces (1,2). At any particular time, as many as 40% of humans may be colonized by culturable S. aureus strains on either nasal or vaginal mucosal surfaces (1,3). The organism may cause relatively benign infections, such as boils, and life threatening infections such as toxic shock syndrome (TSS) (4), scalded skin syndrome (5), necrotizing pneumonia (6-8), and the recently described staphylococcal purpura fulminans (9,10). Antibiotic resistance in S. aureus strains is an ever increasing problem, with recognition of both community-and hospital-associated methicillin-resistant strains (MRSA) (1,2,11). † This work was supported by a research grant from Procter & Gamble, Cincinnati, Ohio and USPHS research grant HL36611 from the National Heart, Lung, and Blood Institute.*To whom correspondence should be addressed. Phone: 612-624-1484; Fax: 612-626-0623; E mail: schli001@umn.edu Recently, two methicillin-resistant, vancomycin-resistant strains were isolated in association with human infections (12,13).The ability of S. aureus to cause serious human diseases depends on production of both cellsurface and secreted virulence factors by the organism (1,4). The cell-surface virulence factors, often referred to as microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) (14), allow the organism to attach to host tissues and in some instances evade the host immune system (15). MSCRAMMs include staphylococcal protein A. The secreted virulence factors, including a large array of exotoxins, allow the organism to gain access to nutrients and avoid the host immune system (4,5,16). For example, the hemolysin family of exotoxins allows the organism both to resist destruction by leukocytes and at the same time gain access to host tissue ...

Staphylococcal toxic shock syndrome toxin 1 (TSST-1) is the cause of menstrual toxic shock syndrome (mTSS) associated with vaginal colonization by Staphylococcus aureus. In this pilot study, we measured TSST-1 and alpha-toxin, another exotoxin, on used tampons from four healthy women with S. aureus on tampons and from two women with tampon-associated mTSS. Tampons from all six women were sectioned into approximately 0.5-cm 3 pieces, some containing menstrual blood and some lacking menstrual blood. The pH of tampon sections with or without menstrual blood was neutral. S. aureus CFU were present in tampon sections at approximately equivalent counts (total counts were 1 ؋ 10 8 to 2 ؋ 10 9 CFU/tampon). TSST-1 (2 to 80 g/tampon) and alpha-toxin (28 to 30 g/tampon) were present only in the sections containing little or no menstrual blood (low hemoglobin density). In the tampons from TSS patients, the cytokine gamma interferon (IFN-␥) was detected only in menstrual-blood-containing sections, whereas the chemokines macrophage inflammatory protein 3␣ and interleukin-8 were detected in all sections. Thus, IFN-␥ was being produced systemically, whereas the chemokines were being produced both locally by epithelial cells and systemically. The data show that S. aureus exotoxins can be identified in tampons ex vivo in sites with low hemoglobin density.

Gram-positive bacteria, such as Staphylococcus aureus, cause serious human illnesses through combinations of surface virulence factors and secretion of exotoxins. Our prior studies using the protein synthesis inhibitor clindamycin and signal transduction inhibitors glycerol monolaurate and α-globin and β-globin chains of hemoglobin indicate that their abilities to inhibit exotoxin production by S. aureus are separable from abilities to inhibit growth of the organism. Additionally, our previous studies suggest that inhibition of exotoxin production, in absence of ability to kill S. aureus and normal flora lactobacilli, will prevent colonization by pathogenic S. aureus, while not interfering with lactobacilli colonization. These disparate activities may be important in development of novel anti-infective agents that do not alter normal flora. We initiated studies to explore the exotoxin-synthesis-inhibition activity of hemoglobin peptides further to develop potential agents to prevent S. aureus infections. We tested synthesized α-globin chain peptides, synthetic variants of α-globin chain peptides, and two human defensins for ability to inhibit exotoxin production without significantly inhibiting S. aureus growth. All of these peptides were weakly or not inhibitory to bacterial growth. However, the peptides were inhibitory to exotoxin production with increasing activity dependent on increasing numbers of positively-charged amino acids. Additionally, the peptides could be immobilized on agarose beads or have amino acid sequences scrambled and still retain exotoxin-synthesis-inhibition. The peptides are not toxic to human vaginal epithelial cells and do not inhibit growth of normal flora L. crispatus. These peptides may interfere with plasma membrane signal transduction in S. aureus due to their positive charges.