Bacterial vaginosis (BV) is a most common microbiological syndrome. Multiplex next-generation sequencing (NGS) or molecular tests allow a complete and accurate vaginal microbiota profiling in order to determine the primary causative agent. Due to the high costs and limited availability of NGS, the multiplex real-time PCR draws more attention. The present study aimed to evaluate the microbial composition and dominant lactobacilli species in non-pregnant women with bacterial vaginosis using a multiplex RT-PCR test and determine its diagnostic significance. In total, 331 women complaining of vaginal discharge were included. BV was confirmed upon clinical examination and Nugent criteria. A real-time PCR test was carried out with a new Femoflor test, which identifies opportunistic bacteria, STD pathogens, and some viruses. According to the results, the rate of lactobacilli is significantly reduced in BV-affected patients when compared to healthy women. Moreover, the rate of L. crispatus significantly decreases, while the rate of L. iners remains high. Among obligate anaerobic bacteria, Gardnerella vaginalis was the most prevalent in women with BV. The Femoflor test demonstrated high sensitivity and specificity for diagnosing BV. Moreover, the test allows the identification of infection in women with intermediate vaginal microbiota, as well as STD pathogens, and viruses. Thus, the application of real-time PCR tests can be effectively used in vaginal microbiota evaluation in women with BV, intermediate vaginal microbiota, and healthy women. In addition, this test may be used as an alternative to the Amsel criteria and Nugent scoring method in diagnosing BV.

BACKGROUND: A microbial imbalance in the vaginal biotope leads to the development of vaginal infections and is associated with urinary tract infections. The ineffectiveness of the therapy and the frequent recurrence of these infections are important reasons for searching for alternative treatment strategies. A progressive solution to this issue has been the use of oral probiotics with a targeted effect on the intestinal and vaginal microflora. AIM: The aim of this study was to evaluate the antagonistic and antibacterial activity of the probiotic strains Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus La-14, which are the components of the oral probiotic, against opportunistic microorganisms of intestinal origin, while assessing the sensitivity of these strains to antimicrobial drugs. MATERIALS AND METHODS: The probiotic strains Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus La-14 included in the oral probiotic Duogynal® were cultivated and the grown colonies were identified using the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry method (Bruker Daltonics GmbH Co., Germany). To study the antagonism of the probiotic strains of lactobacilli and opportunistic pathogenic microorganisms, the following 11 clinical isolates were collected: Escherichia coli (two isolates), Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter asburiae, Proteus mirabilis, Citrobacter freundii, Staphylococcus aureus, Enterococcus faecalis, Streptococcus agalactiae, and Candida albicans. To assess the sensitivity of the probiotic strains to antimicrobial drugs, the Gram Positive AST panel (Autobio Diagnostics Co., Ltd, China) was used. RESULTS: When the microorganisms were co-cultivated with the probiotic lactobacilli (Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus La-14), we noted a decrease in pH and the death of opportunistic microorganisms such as Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter asburiae, Proteus mirabilis, Citrobacter freundii, Staphylococcus aureus, Enterococcus faecalis, and Streptococcus agalactiae, as well as yeast-like fungi Candida albicans. When studying antibiotic resistance, it was found that the both strains of lactobacilli were resistant to ampicillin, vancomycin, daptomycin, clindamycin, linezolid, moxifloxacin, nitrofurantoin, oxacillin, oritavancin, penicillin, rifampin, teicoplanin, tigecycline, trimethoprim/sulfamethoxazole, cefoxitin, ceftarolin, and ciprofloxacin. CONCLUSIONS: The oral probiotic, which contains a combination of two lactobacilli (Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus La-14), has pronounced antagonistic activity against opportunistic microorganisms. These strains are resistant to most antibacterial drugs used in widespread clinical practice, which may be the basis for recommending the use of this probiotic drug together with prescribed antibiotics for prevention of vaginal or intestinal dysbiosis.

Background. Bacterial films are a marker of chronic recurrent infections. Biofilms on mucous membranes block the inflammatory response of the macroorganism, suppressing the activity of immunocytes, and thereby allow microorganisms to reach high concentrations. Currently, research is being conducted to find medications that can act on biofilms. Enzymes, especially their complexes, are substances that can destroy bacterial films. Objective. Еo determine in vitro the sensitivity of bacterial biofilms formed by vaginal microorganisms to the complex of enzymes included in Wobenzym. Materials and methods. The study included 72 clinical isolates of pure microorganism cultures isolated from the vaginal biotope: Gardnerella vaginalis (3), Enterococcus faecalis (9), Escherichia coli (18), Klebsiella pneumoniae (15), Klebsiella aerogenes (3), Lactobacillus crispatus (3), Streptococcus pyogenes (3), Acinetobacter baumanii (3), Staphylococcus aureus (3), Candida albicans (3), Enterococcus faecium (3), Streptococcus agalactiae (3), Lactobacillus acidophilus (3). Bacterial biofilm formation was determined in polystyrene flat-bottom plates using a modified method of Christensen et al. (1985). The tablet form of Wobenzym was used in the study. The tablet shell was washed with saline, the tablet itself was dissolved in 10 ml of 0.9% NaCl and used for in vitro studies. The result was determined using a reader on a spectrophotometer to determine the optical density (OD) of the formed biofilm. It was believed that the drug acted on the bacterial film, reducing the OD by more than three times. Results. An in vitro study revealed clinical isolates of bacteria that formed biofilms of varying severity. Of the 72 clinical bacterial isolates, 38 formed biofilms. A pronounced effect of the complex of enzymes included in Wobenzym on biofilms formed by microorganisms such as A. baumanii, S. aureus, G. vaginalis and E. faecalis was noted. Conclusion. Wobenzym has an effective destructive effect on biofilms formed by various microorganisms, including G. vaginalis, common causative agents of bacterial vaginosis, as well as staphylococci and enterococci, causative agents of aerobic (nonspecific) vaginitis. Conclusion. The drug Wobenzym has an effective destructive effect on biofilms formed by various microorganisms, including Gardnerella vaginalis, common causative agents of bacterial vaginosis, as well as staphylococci and enterococci, causative agents of aerobic (nonspecific vaginitis).