The group of experts representing the Polish Gynecologic Society has issued this statement based on the review of available literature on the potential benefits of the use of Macmiror Complex 500 in obstetrical and gynecologic practice. Mixed Vaginitis (MV) eg. the vaginal infection caused by at least two out of the triad of pathogens (fungi, bacteria and Trichomonas Vaginalis [TV]), constitutes the type of vaginitis which is underestimated as for its prevalence. Mixed pathogens are responsible for as much as one third of all vaginal infections. Macmiror Complex 500 contains two active ingredients: nifuratel and nystatin. Macmiror Complex 500 affects all common causes of vulvovaginitis, i.e. bacteria, yeasts and TV. At the same time, it is not effective against Lactobacillus spp., which is a clear advantage in the treatment of vaginal infections. The antibacterial spectrum of nifuratel includes aerobic and anaerobic bacteria. Moreover nifuratel is effective against Chlamydia trachomatis and Mycoplasma spp., it has an anti-trichomonal effect comparable to metranidazole and shows certain activity against Candida spp. Nystatin is effective against Candida albicans and is even very effective against Candida glabrata which is usually more resistant to imidazole antifungal agents. Nystatin's importance is rising due to the current increase of candidoses caused by non-albicans types. This increase is especially perceptible in recurring candidoses. The review of the available literature on the effectiveness of Macmiror Complex 500 in the OB/GYN practice leads to the following conclusions: the exeptionally broad antibacterial and antifungal and trichomonicidal activity of this formulation makes it a drug of choice in cases where MV is suspected. The possibility to treat both partners, favorable safety profile in pregnant patients and the availability of both vaginal ovules and the cream with applicator makes this drug an effective and suitable treatment option in obstetrical and gynecologic practice.

Menstrual toxic shock syndrome (TSS) is a serious illness that afflicts women of premenopausal age worldwide and arises from vaginal infection by Staphylococcus aureus and concurrent production of toxic shock syndrome toxin-1 (TSST-1). Studies have illustrated the capacity of lactobacilli to reduce S. aureus virulence, including the capacity to suppress TSST-1. We hypothesized that an aberrant microbiota characteristic of pathogenic bacteria would induce the increased production of TSST-1 and that this might represent a risk factor for the development of TSS. A S. aureus TSST-1 reporter strain was grown in the presence of vaginal swab contents collected from women with a clinically healthy vaginal status, women with an intermediate status, and those diagnosed with bacterial vaginosis (BV). Bacterial supernatant challenge assays were also performed to test the effects of aerobic vaginitis (AV)-associated pathogens toward TSST-1 production. While clinical samples from healthy and BV women suppressed toxin production, in vitro studies demonstrated that Streptococcus agalactiae and Enterococcus spp. significantly induced TSST-1 production, while some Lactobacillus spp. suppressed it. The findings suggest that women colonized by S. aureus and with AV, but not BV, may be more susceptible to menstrual TSS and would most benefit from prophylactic treatment.

The vagina is a complex biocenosis where many micro-organisms coexist and colonize it. The dominant colonizing bacteria of a healthy individual is of the genus "lactobacillus". It is also called Doderlein's bacillus and determines the vaginal microbial balance through the production of lactic acid, hydrogen peroxide, biosurfactants, bacteriocines and modify the competition of pathogens for adhesion to the vaginal epithelial cells. Through these mechanisms the lactobacilli block the growth and development of other vaginal pathogenic microbial species and also inhibit the colonization of some other microorganisms imported from outside. Because of these potential therapeutic properties, the lactobacilli are used as effective medical agents for prophylaxys and therapy to restore the physiological balance in the vaginal eco-system. According to our studies and gained clinical experience the etiological antibacterial treatment is not always sufficient to restore the normal vaginal flora. The complete recovery of the vaginal flora could be reached using probiotics that are applied locally and per os. GynOphilus is a new vaginal probiotic product containing one type of the genus "Lactobacillus": Lactobacillus casei var rhamnosus. GynOphilus restores the physiological balance of the vaginal flora and reduces the risk of recurrent infection. The product is applied intravaginal and interacts locally, inhibits the growth of the most common vaginal pathogens: Gardnerella vaginalis u Candida albicans.

Bacterial vaginosis is uncommon in women who are virgins. We estimated effects of sexual debut on vaginal bacterial colonization.Women who were virgins and aged 18-22 years enrolled in a study of human papillomavirus acquisition were followed every 4 months for up to 2 years. Vaginal swabs from before and after sexual debut or two independent visits for those remaining virgins were tested by quantitative polymerase chain reaction for Lactobacillus crispatus, Lactobacillus jensenii, Lactobacillus iners, Gardnerella vaginalis, and the bacterial vaginosis-associated species Atopobium vaginae, Megasphaera species, Leptotrichia species, Sneathia species, and bacterial vaginosis-associated bacterium-1, -2, and -3.We evaluated 97 women: 71 who became sexually active and 26 who remained virgins. At first sampling, 22 of 26 (85%) women who remained virgins were colonized with Lactobacillus species compared with 22 of 26 (85%) at follow-up (P>.99). G vaginalis was present in 12 of 26 (46%) initially and 11 of 26 (42%) at follow-up (P>.99). Among women who became sexually active, colonization with Lactobacillus species remained stable: 65 of 71 (92%) compared with 66 of 71 (93%) (P>.99), whereas colonization with G vaginalis increased (28 of 71 [39%] compared with 40 of 71 [56%]; P=.02). Among women who did not initiate sexual activity during the study, two of 26 (8%) had any bacterial vaginosis-associated species detected at both the first and second visits (P>.99). Among women who became sexually active during the study, 15 of 71 (21%) were colonized with bacterial vaginosis-associated species initially compared with 13 of 71 (18%) after sexual debut (P=.77).Among women who were virgins, vaginal colonization with bacterial vaginosis-associated bacterial species is uncommon and does not change after sexual debut.

Bacterial vaginosis (BV) is the most commonly treated female reproductive tract affliction, characterized by the displacement of healthy lactobacilli by an overgrowth of pathogenic bacteria. BV can contribute to pathogenic inflammation, preterm birth, and susceptibility to sexually transmitted infections. As the bacteria responsible for BV pathogenicity and their interactions with host immunity are not understood, we sought to evaluate the effects of BV-associated bacteria on reproductive epithelia. Here we have characterized the interaction between BV-associated bacteria and the female reproductive tract by measuring cytokine and defensin induction in three types of FRT epithelial cells following bacterial inoculation. Four BV-associated bacteria were evaluated alongside six lactobacilli for a comparative assessment. While responses differed between epithelial cell types, our model showed good agreement with clinical BV trends. We observed a distinct cytokine and human β-defensin 2 response to BV-associated bacteria, especially Atopobium vaginae, compared to most lactobacilli. One lactobacillus species, Lactobacillus vaginalis, induced an immune response similar to that elicited by BV-associated bacteria, stimulating significantly higher levels of cytokines and human β-defensin 2 than other lactobacilli. These data provide an important prioritization of BV-associated bacteria and support further characterization of reproductive bacteria and their interactions with host epithelia. Additionally, they demonstrate the distinct immune response potentials of epithelial cells from different locations along the female reproductive tract.

The aim of our study was to study the relationship of blastospores and pseudohyphae in Papanicolaou (Pap) smears and Nugent scores for bacterial vaginosis (BV).A total of 471 Pap smears with Candida albicans were reviewed. The presence of blastospores and pseudohyphae was established. The Pap smears were restained with the Gram stain method to evaluate the bacterial flora according to the Nugent scoring system.Of the 471 Pap smears, blastospores and pseudohyphae were observed in 62.8% (296/471) and 37.2% (175/471) of the smears, and displayed symptoms in 4.4% (13/296) and 43.4% (76/175), respectively. A significant difference was found between these 2 groups (p < 0.0001). A positive BV Nugent score (≥ 7) was found in 22.1% (104/471) of the C. albicans cases. Blastospores and pseudohyphae with BV were 14.2% (42/296) and 35.4% (62/175), respectively. These high Nugent scores indicate statistically significant differences (p < 0.0001).C. albicans and BV can coexist. The presence of blastospores in these C. albicans cases was negatively related to symptoms.

We demonstrate the feasibility of using qPCR on DNA extracted from vaginal Gram stain slides to estimate the presence and relative abundance of specific bacterial pathogens. We first tested Gram stained slides spiked with a mix of 10(8) cfu/ml of Escherichia coli and 10(5) cfu/ml of Lactobacillus acidophilus. Primers were designed for amplification of total and species-specific bacterial DNA based on 16S ribosomal gene regions. Sample DNA was pre-amplified with nearly full length 16S rDNA ribosomal gene fragment, followed by quantitative PCR with genera and species-specific 16S rDNA primers. Pre-amplification PCR increased the bacterial amounts; relative proportions of Escherichia coli and Lactobacillus recovered from spiked slides remained unchanged. We applied this method to forty two archived Gram stained slides available from a clinical trial of cerclage in pregnant women at high risk of preterm birth. We found a high correlation between Nugent scores based on bacterial morphology of Lactobacillus, Gardenerella and Mobiluncus and amounts of quantitative PCR estimated genus specific DNA (rrn copies) from Gram stained slides. Testing of a convenience sample of eight paired vaginal swabs and Gram stains freshly collected from healthy women found similar qPCR generated estimates of Lactobacillus proportions from Gram stained slides and vaginal swabs. Archived Gram stained slides collected from large scale epidemiologic and clinical studies represent a valuable, untapped resource for research on the composition of bacterial communities that colonize human mucosal surfaces.

Some vaginal bacterial communities are thought to prevent infection by sexually transmitted organisms. Prior work demonstrated that the vaginal microbiota of reproductive-age women cluster into 5 types of bacterial communities; 4 dominated by Lactobacillus species (L. iners, L. crispatus, L. gasseri, L. jensenii) and 1 (termed community state type (CST) IV) lacking significant numbers of lactobacilli and characterized by higher proportions of Atopobium, Prevotella, Parvimonas, Sneathia, Gardnerella, Mobiluncus, and other taxa. We sought to evaluate the relationship between vaginal bacterial composition and Trichomonas vaginalis.Self-collected vaginal swabs were obtained cross-sectionally from 394 women equally representing 4 ethnic/racial groups. T. vaginalis screening was performed using PCR targeting the 18S rRNA and β-tubulin genes. Vaginal bacterial composition was characterized by pyrosequencing of barcoded 16S rRNA genes. A panel of 11 microsatellite markers was used to genotype T. vaginalis. The association between vaginal microbiota and T. vaginalis was evaluated by exact logistic regression.T. vaginalis was detected in 2.8% of participants (11/394). Of the 11 T. vaginalis-positive cases, 8 (72%) were categorized as CST-IV, 2 (18%) as communities dominated by L. iners, and 1 (9%) as L. crispatus-dominated (P = 0.05). CST-IV microbiota were associated with an 8-fold increased odds of detecting T. vaginalis compared with women in the L. crispatus-dominated state (OR: 8.26, 95% CI: 1.07-372.65). Seven of the 11 T. vaginalis isolates were assigned to 2 genotypes.T. vaginalis was associated with vaginal microbiota consisting of low proportions of lactobacilli and high proportions of Mycoplasma, Parvimonas, Sneathia, and other anaerobes.

Vulvovaginal candidiasis (VVC) is the second most common cause of vaginitis after bacterial vaginosis, and it is diagnosed in up to 40% of women with vaginal complaints in the primary care setting. Among Candida spp., Candida albicans is the most common infectious agent. The treatment of choice for uncomplicated VVC is achieved with single-dose or short-course therapy in over 90% of cases. Several topical and oral drugs are available, without evidence for superiority of any agent or route of administration. In any case, most classic treatments are unable to significantly offer a protection against possible recurrences. In recent years, probiotics are emerging as a new strategy to counteract VVC. In fact, they are well known for their ability to lower intravaginal pH, thus establishing a barrier effect against many types of yeasts. Some strains are also able to exert additional and more focused antagonistic activities mediated by specific molecules such as hydrogen peroxide and bacteriocins. For example, Lactobacillus fermentum LF5 (CNCM I-789) was successfully tested in 4 human trials involving a total of 340 women reporting VVC at enrollment. In any case, the way used to deliver probiotics to the vaginal environment represents a crucial point. The aim of this work was to first select 1 or more probiotic strains in vitro with an antagonistic activity on Candida yeasts and then to perform an in vivo human pilot study using an association of the most promising and active bacteria.For this purpose, 2 probiotic strains Probiotical S.p.A (Italy) were selected based on their strong in vitro inhibition activity toward 4 particular Candida species, namely C. albicans, Candida glabrata, Candida parapsilosis, and Candida krusei and subsequently tested in a human intervention pilot trial involving 30 women with VVC. The probiotics used, L. fermentum LF10 (DSM 19187) and Lactobacillus acidophilus LA02 (DSM 21717), were administered by means of slow release effervescent vaginal tablets (ActiCand 30 product). The main endpoint was the assessment of the establishment and maintenance of a barrier effect against Candida yeasts in women suffering from VVC. Thirty female subjects who were diagnosed with VVC by both microscopic examination and yeast culture were enrolled in the study and directed to apply a vaginal tablet once a day for 7 consecutive nights, followed by 1 tablet every 3 nights for a further 3-week application (acute phase) and, finally, 1 tablet per week to maintain a long-term vaginal colonization against possible recurrences. A medical examination of each patient was performed at enrollment (d₀), at the end of the first 4 weeks of treatment (d₂₈), and at the end of the second month of relapse prevention (d₅₆). The visual and microscopic examination was always accompanied by microbiological analyses of vaginal swabs to assess the presence of Candida. A statistical comparison was made between d₂₈, or d₅₆, and d0, and between d₅₆ and d₂₈ to quantify the efficacy against possible recurrences.The administration of the product ActiCand 30 was able to significantly solve Candida yeast symptoms after 28 days in 26 patients out of 30 (corresponding to 86.6%, P<0.001). At the end of the second month, recurrences were recorded, albeit not particularly serious, in only 3 out of 26 patients (11.5%, P=0.083) who were found to have fully healed at the end of the first month of treatment. This is a further confirmation of the long-term barrier effect exerted by the product.VVC has a very high incidence as 70% to 75% of women report at least 1 episode during the life. Many treatments are currently available but, despite a relatively high effectiveness in the relief of symptoms typically associated with acute infections, they are generally unable to offer a long-term protective barrier against possible recurrences. This study demonstrated the ability of ActiCand 30 to not only solve Candida infections in a very high percentage of women, but also to exert a long-term physiological defense due to the colonization of vaginal microbiota and adhesion of the mucosa to the epithelial cells. The special formulation of ActiCand 30, consisting of slow release effervescent vaginal tablets, is able to mediate 2 types of barrier effects, the first represented by the formation of an anaerobic environment due to the release of CO₂ and the second guaranteed by the colonization and adhesion to the vaginal epithelium of the 2 probiotics L. fermentum LF10 and L. acidophilus LA02.

Bacterial vaginosis (BV), characterized by a shift of the vaginal microbiota from a Lactobacillus-dominated community to a dense biofilm containing a complex mixture of organisms, is an important risk factor in poor reproductive health outcomes. The Nugent score, based on Gram stain, is used to diagnose BV and Gardnerella vaginalis abundance in the sample is one factor determining Nugent score. A high Nugent score is indicative of BV but does not always correspond to the presence of clinical symptoms. G. vaginalis is recognized as a heterogeneous group of organisms, which can also be part of the normal, healthy vaginal microbiome. In addition, asymptomatic BV and non-Gardnerella types of BV are being recognized. In an attempt to resolve the heterogeneous group of G. vaginalis, a phylogenetic tree of cpn60 universal target sequences from G. vaginalis isolates was constructed that indicates the existence of four subgroups of G. vaginalis. This subdivision, supported by whole genome similarity calculation of representative strains using JSpecies, demonstrates that these subgroups may represent different species. The cpn60 subgroupings did not correspond with the Piot biotyping scheme, but did show consistency with ARDRA genotyping and sialidase gene presence. Isolates from all four subgroups produced biofilm in vitro. We also investigated the distribution of G. vaginalis subgroups in vaginal samples from Kenyan women with Nugent scores consistent with BV, Intermediate and Normal microbiota (n = 44). All subgroups of G. vaginalis were detected in these women, with a significant difference (z = -3.372, n = 39, p = 0.001) in frequency of G. vaginalis subgroup B between BV and Normal groups. Establishment of a quantifiable relationship between G. vaginalis subgroup distribution and clinical status could have significant diagnostic implications.