title: Isolation of Vaginal Lactobacilli and Characterization of Anti-Candida Activity.
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.
authors.affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.
authors.forename: Carola
authors.initials: C
authors.lastname: Parolin
authors.affiliation: Microbiology, DIMES, University of Bologna, Bologna, Italy.
authors.forename: Antonella
authors.initials: A
authors.lastname: Marangoni
authors.affiliation: Centre of Foodomics, Department of Agro-Food Science and Technology, University of Bologna, Bologna, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Laghi
authors.affiliation: Microbiology, DIMES, University of Bologna, Bologna, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Foschi
authors.affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.
authors.forename: Rogers Alberto
authors.initials: RA
authors.lastname: Ñahui Palomino
authors.affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.
authors.forename: Natalia
authors.initials: N
authors.lastname: Calonghi
authors.affiliation: Microbiology, DIMES, University of Bologna, Bologna, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Cevenini
authors.affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.
authors.forename: Beatrice
authors.initials: B
authors.lastname: Vitali
mesh_terms: D000293:Adolescent; D000328:Adult; D000935:Antifungal Agents / Q000378:metabolism; D001483:Base Sequence; D002175:Candida / Q000502:physiology; D002181:Candidiasis, Vulvovaginal / Q000382:microbiology*; D005260:Female; D006367:HeLa Cells / Q000382:microbiology; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007778:Lactobacillus / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D009682:Magnetic Resonance Spectroscopy; D055432:Metabolomics; D064307:Microbiota / Q000502:physiology; D008875:Middle Aged; D008969:Molecular Sequence Data; D014621:Vagina / Q000382:microbiology*; D055815:Young Adult
chemical_list: D000935:Antifungal Agents; D006861:Hydrogen Peroxide