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Isolation of Vaginal Lactobacilli and Characterization of Anti-Candida Activity.

Parolin C, Marangoni A, Laghi L, Foschi C, Ñahui Palomino RA, Calonghi N, Cevenini R, Vitali B - PLoS ONE (2015)

Bottom Line: Most Lactobacillus strains significantly reduced C. albicans adhesion through all mechanisms.In particular, L. crispatus BC2, L. gasseri BC10 and L. gasseri BC11 appeared to be the most active strains in reducing pathogen adhesion, as their effects were mediated by both cells and supernatants.Our results are prerequisites for the development of new therapeutic agents based on probiotics for prophylaxis and adjuvant therapy of Candida infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.

ABSTRACT
Healthy vaginal microbiota is dominated by Lactobacillus spp., which form a critical line of defence against pathogens, including Candida spp. The present study aims to identify vaginal lactobacilli exerting in vitro activity against Candida spp. and to characterize their antifungal mechanisms of action. Lactobacillus strains were isolated from vaginal swabs of healthy premenopausal women. The isolates were taxonomically identified to species level (L. crispatus B1-BC8, L. gasseri BC9-BC14 and L. vaginalis BC15-BC17) by sequencing the 16S rRNA genes. All strains produced hydrogen peroxide and lactate. Fungistatic and fungicidal activities against C. albicans, C. glabrata, C. krusei, C. tropicalis, C. parapsilosis and C. lusitaniae were evaluated by broth micro-dilution method. The broadest spectrum of activity was observed for L. crispatus BC1, BC4, BC5 and L. vaginalis BC15, demonstrating fungicidal activity against all isolates of C. albicans and C. lusitaniae. Metabolic profiles of lactobacilli supernatants were studied by 1H-NMR analysis. Metabolome was found to be correlated with both taxonomy and activity score. Exclusion, competition and displacement experiments were carried out to investigate the interference exerted by lactobacilli toward the yeast adhesion to HeLa cells. Most Lactobacillus strains significantly reduced C. albicans adhesion through all mechanisms. In particular, L. crispatus BC2, L. gasseri BC10 and L. gasseri BC11 appeared to be the most active strains in reducing pathogen adhesion, as their effects were mediated by both cells and supernatants. Inhibition of histone deacetylases was hypothesised to support the antifungal activity of vaginal lactobacilli. Our results are prerequisites for the development of new therapeutic agents based on probiotics for prophylaxis and adjuvant therapy of Candida infection.

No MeSH data available.


Related in: MedlinePlus

Interference of vaginal lactobacilli with C. albicans adhesion to HeLa cells.Exclusion (A), competition (B) and displacement (C) experiments were performed. The results were expresses as percentages of adherent yeasts per HeLa cell and compared with adhesion without lactobacilli (control value). The control value was taken as 100% of adhesion (black bars). White and grey bars show the adhesion of C. albicans in presence of Lactobacillus cells and supernatants, respectively. Statistical significance was determined at P < 0.05*, P < 0.01** and P < 0.001***. Error bars represent SEM.
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pone.0131220.g003: Interference of vaginal lactobacilli with C. albicans adhesion to HeLa cells.Exclusion (A), competition (B) and displacement (C) experiments were performed. The results were expresses as percentages of adherent yeasts per HeLa cell and compared with adhesion without lactobacilli (control value). The control value was taken as 100% of adhesion (black bars). White and grey bars show the adhesion of C. albicans in presence of Lactobacillus cells and supernatants, respectively. Statistical significance was determined at P < 0.05*, P < 0.01** and P < 0.001***. Error bars represent SEM.

Mentions: To verify the antagonist effect of the vaginal lactobacilli against C. albicans, the influence of CP and their respective CFS on the adhesion capacity of the yeast to HeLa cells was investigated (Fig 3). Three mechanisms of inhibition were examined: exclusion (Fig 3A), competition (Fig 3B) and displacement (Fig 3C). Ten strains (BC1, BC2, BC5, BC7, BC8, BC9, BC10, BC11, BC12 and BC16) significantly reduced the adhesion of C. albicans through all three mechanisms. The inhibitory effect was exerted in some cases by CP and in other cases by CFS. In particular, the interference by L. crispatus BC2, L. gasseri BC10 and L. gasseri BC11 was mediated by both CP and CFS, suggesting that these strains were the most active in terms of inhibition of the pathogen adhesion. Interestingly, BC2, BC10 and BC11 were not the most adhesive strains, as shown by the Fig 2. Only three strains did not exercise any effect on C. albicans adhesion: L. gasseri BC13, L. gasseri BC14 and L. vaginalis BC17. The remaining strains showed an intermediate behaviour exerting inhibition through one or two mechanisms.


Isolation of Vaginal Lactobacilli and Characterization of Anti-Candida Activity.

Parolin C, Marangoni A, Laghi L, Foschi C, Ñahui Palomino RA, Calonghi N, Cevenini R, Vitali B - PLoS ONE (2015)

Interference of vaginal lactobacilli with C. albicans adhesion to HeLa cells.Exclusion (A), competition (B) and displacement (C) experiments were performed. The results were expresses as percentages of adherent yeasts per HeLa cell and compared with adhesion without lactobacilli (control value). The control value was taken as 100% of adhesion (black bars). White and grey bars show the adhesion of C. albicans in presence of Lactobacillus cells and supernatants, respectively. Statistical significance was determined at P < 0.05*, P < 0.01** and P < 0.001***. Error bars represent SEM.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4476673&req=5

pone.0131220.g003: Interference of vaginal lactobacilli with C. albicans adhesion to HeLa cells.Exclusion (A), competition (B) and displacement (C) experiments were performed. The results were expresses as percentages of adherent yeasts per HeLa cell and compared with adhesion without lactobacilli (control value). The control value was taken as 100% of adhesion (black bars). White and grey bars show the adhesion of C. albicans in presence of Lactobacillus cells and supernatants, respectively. Statistical significance was determined at P < 0.05*, P < 0.01** and P < 0.001***. Error bars represent SEM.
Mentions: To verify the antagonist effect of the vaginal lactobacilli against C. albicans, the influence of CP and their respective CFS on the adhesion capacity of the yeast to HeLa cells was investigated (Fig 3). Three mechanisms of inhibition were examined: exclusion (Fig 3A), competition (Fig 3B) and displacement (Fig 3C). Ten strains (BC1, BC2, BC5, BC7, BC8, BC9, BC10, BC11, BC12 and BC16) significantly reduced the adhesion of C. albicans through all three mechanisms. The inhibitory effect was exerted in some cases by CP and in other cases by CFS. In particular, the interference by L. crispatus BC2, L. gasseri BC10 and L. gasseri BC11 was mediated by both CP and CFS, suggesting that these strains were the most active in terms of inhibition of the pathogen adhesion. Interestingly, BC2, BC10 and BC11 were not the most adhesive strains, as shown by the Fig 2. Only three strains did not exercise any effect on C. albicans adhesion: L. gasseri BC13, L. gasseri BC14 and L. vaginalis BC17. The remaining strains showed an intermediate behaviour exerting inhibition through one or two mechanisms.

Bottom Line: Most Lactobacillus strains significantly reduced C. albicans adhesion through all mechanisms.In particular, L. crispatus BC2, L. gasseri BC10 and L. gasseri BC11 appeared to be the most active strains in reducing pathogen adhesion, as their effects were mediated by both cells and supernatants.Our results are prerequisites for the development of new therapeutic agents based on probiotics for prophylaxis and adjuvant therapy of Candida infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.

ABSTRACT
Healthy vaginal microbiota is dominated by Lactobacillus spp., which form a critical line of defence against pathogens, including Candida spp. The present study aims to identify vaginal lactobacilli exerting in vitro activity against Candida spp. and to characterize their antifungal mechanisms of action. Lactobacillus strains were isolated from vaginal swabs of healthy premenopausal women. The isolates were taxonomically identified to species level (L. crispatus B1-BC8, L. gasseri BC9-BC14 and L. vaginalis BC15-BC17) by sequencing the 16S rRNA genes. All strains produced hydrogen peroxide and lactate. Fungistatic and fungicidal activities against C. albicans, C. glabrata, C. krusei, C. tropicalis, C. parapsilosis and C. lusitaniae were evaluated by broth micro-dilution method. The broadest spectrum of activity was observed for L. crispatus BC1, BC4, BC5 and L. vaginalis BC15, demonstrating fungicidal activity against all isolates of C. albicans and C. lusitaniae. Metabolic profiles of lactobacilli supernatants were studied by 1H-NMR analysis. Metabolome was found to be correlated with both taxonomy and activity score. Exclusion, competition and displacement experiments were carried out to investigate the interference exerted by lactobacilli toward the yeast adhesion to HeLa cells. Most Lactobacillus strains significantly reduced C. albicans adhesion through all mechanisms. In particular, L. crispatus BC2, L. gasseri BC10 and L. gasseri BC11 appeared to be the most active strains in reducing pathogen adhesion, as their effects were mediated by both cells and supernatants. Inhibition of histone deacetylases was hypothesised to support the antifungal activity of vaginal lactobacilli. Our results are prerequisites for the development of new therapeutic agents based on probiotics for prophylaxis and adjuvant therapy of Candida infection.

No MeSH data available.


Related in: MedlinePlus