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Survival and biofilm formation by Group B streptococci in simulated vaginal fluid at different pHs
Publication . Borges, Sandra; Silva, Joana; Teixeira, Paula
The Group B Streptococcus (GBS, Streptococcus agalactiae) is an important cause of neonatal and maternal infection. GBS is a commensal organism of the lower gastrointestinal and vaginal tract. A frequent mode of neonatal infection is vertical transmission from pregnant women to their foetus or neonate. The aim of this study was to evaluate the survival and biofilm production of 10 GBS strains in simulated vaginal fluid at pH 4.2, 5.5 and 6.5. GBS survived longer at higher pH than at normal vaginal pH. At pH 4.2, with the exception of two isolates that were recovered up to 48 and 72 h, viable GBS numbers declined below the limit of detection by 24 h. At higher pH, GBS survived between 3 and 15 days. All isolates investigated were biofilm producers but biofilm production was greater in tryptone soy broth compared to simulated vaginal fluid. The quantity of biofilm produced increased with the rise in the pH. This study suggests that high vaginal pH may influence both GBS survival and biofilm production and thus could be a risk factor for GBS infection.
Evaluation of characteristics of Pediococcus spp. to be used as a vaginal probiotic
Publication . Borges, S.; Barbosa, J.; Silva, Joana; Teixeira, Paula
The aim of our research was to select, identify and characterize an
isolate of lactic acid bacteria to be considered as a vaginal probiotic.
Methods and Results: Thirty-five isolates of Pediococcus spp. showed
bacteriocinogenic activity against Listeria monocytogenes and the ability to
survive in simulated vaginal fluid (SVF) at pH 4!2. One isolate of Pediococcus
spp. was selected and characterized to evaluate its safety before the use as
vaginal probiotic. Pediococcus pentosaceus SB83 did not show the presence of
virulence factors such as the production of gelatinase, lipase and DNase,
haemolytic activity, nor the presence of virulence genes (genes esp, agg, gelE,
efaAfm, efaAfs, cylA, cylB and cylM). Pediococcus pentosaceus SB83 was
considered sensitive to chloramphenicol, gentamicin, streptomycin, kanamycin,
erythromycin and ampicillin. This strain was considered resistant to
tetracycline and vancomycin. Pediococcus pentosaceus SB83 was a biofilm
producer at different pH values (4!2, 5!5 and 6!5) in SVF and in de Man,
Rogosa and Sharpe medium.
Conclusions: The in vitro results provide a basis for the use of P. pentosaceus
SB83 as a vaginal probiotic, to prevent colonization by L. monocytogenes in
pregnant women.
Significance and Impact of Study: The application of vaginal probiotics could
have the potential for preventing vaginal infections and consequently reduce
abortion and neonatal infections.
Effects of processing and storage on Pediococcus pentosaceus SB83 in vaginal formulations: lyophilized powder and tablets
Publication . Borges, Sandra; Costa, Paulo; Silva, Joana; Teixeira, Paula
Vaginal probiotics have an important role in preventing the colonization of the vagina by pathogens. This study aimed to investigate different formulations with Pediococcus pentosaceus SB83 (lyophilized powder and tablets with and without retarding polymer) in order to verify its stability and antilisterial activity after manufacture and during storage. The bacteriocinogenic activity of P. pentosaceus SB83 against Listeria monocytogenes was evaluated in simulated vaginal fluid. Suspension of Pediococcus pentosaceus SB83 reduced the pathogen only after 2 h and the lyophilized bacteria after 24 h of contact, and, in the tablets, P. pentosaceus SB83 lost the antimicrobial activity. The pH of simulated vaginal fluid decreased for all the tested conditions. As lyophilized powder demonstrated better results concerning antimicrobial activity, this formulation was selected to evaluate the antilisterial activity during the 12 months of storage. During storage at room temperature, lyophilized bacteria totally inhibited the pathogen only until one month of storage. At 4 degrees C, P. pentosaceus SB83 showed antimicrobial activity during all the time of storage investigated. Therefore, the better formulation of P. pentosaceus SB83 is the lyophilized powder stored at 4 degrees C, which may be administered intravaginally as a washing solution.
Characterization of a bacteriocin of pediococcus pentosaceus SB83 and its potential for vaginal application
Publication . Borges, Sandra; Barbosa, Joana; Silva, Joana; Teixeira, Paula
Listeria monocytogenes, the responsible agent for listeriosis, is one of several microorganisms which can be transmitted from mother to fetus/neonate by vertical transmission. The aim of this study was to characterize a bacteriocin produced by Pediococcus pentosaceus SB83 for vaginal application, to prevent colonization of pregnant women with L. monocytogenes. Bacteriocin SB83 showed inhibitory activity against L. monocytogenes (serotypes 1/2a 1/2b and 4b) but did not inhibit vaginal lactic acid bacteria. Additionally, bacteriocin SB83 inhibited the growth of Enterococcus faecalis, Enterococcus faecium and Bacillus subtilis. Bacteriocin SB83 was produced at maximum levels between 24 h and 36 h of growth (6400 AU/mL against serotype 1/2b and 4b, 3200 AU/mL against serotype 1/2a). Significant reduction in antimicrobial activity was observed after treatment of cell-free supernatants with proteinase K, pepsin, papain and trypsin. Bacteriocin SB83 was resistant to treatments with Tween 80, Tween 20, Triton X-100, SDS and EDTA. This bacteriocin was stable at pH values between 4 and 6.5; a decrease in activity was observed at pH 2 and above 8. A decrease in antibacterial activity was recorded at 100 °C and 121 °C. In the range 4 °C to 80 °C, antimicrobial activity was not affected. The activity of bacteriocin did not change in simulated vaginal fluid (pH 4.2, 5.5 and 6.5). The molecular size of bacteriocin SB83 was between 3.5 and 6.5 kDa.In conclusion, the bacteriocin produced by P. pentosaceus SB83 has been proved to be resistant to several conditions, including conditions in the vaginal tract (pH and components of vaginal fluid).
The role of lactobacilli and probiotics in maintaining vaginal health
Publication . Borges, Sandra; Silva, Joana; Teixeira, Paula
Background
The vaginal microbiota of healthy women consists typically of a diversity of anaerobic and aerobic microorganisms. Lactobacilli are the most prevalent and often numerically dominant microorganisms and are relevant as a barrier to infection. The capacity of lactobacilli to adhere and compete for adhesion sites in the vaginal epithelium and the capacity to produce antimicrobial compounds (hydrogen peroxide, lactic acid, bacteriocin-like substances), are important in the impairment of colonization by pathogens.
Objective
This review summarizes the role of lactic acid bacteria in preventing illness of the host, including bacterial vaginosis, yeast vaginitis, urinary tract infection and sexually transmitted diseases.
Conclusions
The administration of probiotics that colonize the vaginal tract can be important in maintaining a normal urogenital health and also to prevent or treat infections.
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Funding agency
Fundação para a Ciência e a Tecnologia
Funding programme
SFRH
Funding Award Number
SFRH/BD/45496/2008