Abstract
Vulvovaginal candidiasis is the second most common vaginal infection caused by drug-resistant Candida species that affects about 70–75% of reproductive age group women across the globe. As current-day antifungal drugs are ineffective against the biofilms formed by the drug-resistant Candida strains, several natural compounds and antagonistic microbes are being explored as alternative antifungal agents. In the present study, we investigated the anti-biofilm activity of Cell-Free Supernatant (CFS) extracted from the commercially available probiotics VSL-3 against the biofilms of Candida species and also evaluated their efficacy in curbing the yeast-to-hyphal transition. Various methodologies like crystal violet staining and scanning electron microscopy were used to study the effect of CFS against the biofilms formed by the species. The ability of CFS to interfere with yeast to hyphal transition in Candida was studied by colony morphology assay and visually confirmed with phase contrast microscopy. The potential of the CFS of the probiotics was also evaluated using goat buccal tissue, a novel ex-vivo model that mimics the vaginal environment. Moreover, the supernatant extracted from VSL-3 had the ability to down-regulate the expression of virulence genes of Candida from the biofilm formed over the ex-vivo model. These results emphasize the anti-fungal and anti-infective properties of the CFS of VSL-3 against drug-resistant Candida strains causing vulvovaginal candidiasis.
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References
Arendrup MC, Patterson TF (2017) Multidrug-resistant Candida: epidemiology, molecular mechanisms, and treatment. J Infect Dis 216(suppl_3):S445–S451. https://doi.org/10.1093/infdis/jix131
Arthur J, Gharaibeh R, Uronis J et al (2013) VSL#3 probiotic modifies mucosal microbial composition but does not reduce colitis-associated colorectal cancer. Sci Rep 3:2868. https://doi.org/10.1038/srep02868
Barousse MM, Steele C, Dunlap K, Espinosa T, Boikov D, Sobel JD, Fidel Jr PL (2001) Growth inhibition of Candida albicans by human vaginal epithelial cells. J Infect Dis 184(11):1489–1493. https://doi.org/10.1086/324532
Bhattacharya S, Sae-Tia S, Fries BC (2020) Candidiasis and mechanisms of antifungal resistance. Antibiotics 9(6):312. https://doi.org/10.3390/antibiotics9060312
Chandra J, Kuhn DM, Mukherjee PK, Hoyer LL, McCormick T, Ghannoum MA (2001) Biofilm formation by the fungal pathogen Candida albicans: development, architecture, and drug resistance. J Bacteriol 183(18):5385–5394. https://doi.org/10.1128/JB.183.18.5385-5394.2001
Chew SY, Cheah YK, Seow HF, Sandai D, Than LT (2015) In vitro modulation of probiotic bacteria on the biofilm of Candida glabrata. Anaerobe 34:132–138. https://doi.org/10.1016/j.anaerobe.2015.05.009
Cinque B, La Torre C, Lombardi F, Palumbo P, Van der Rest M et al (2016) Production conditions affect the in vitro anti-tumoral effects of a high concentration multi-strain probiotic preparation. PLoS ONE 11(9):0163216. https://doi.org/10.1371/journal.pone.0163216
Cotter PD, Hill C, Ross RP (2005) Bacteriocins: developing innate immunity for food. Nat Rev Microbiol 3(10):777–788. https://doi.org/10.1038/nrmicro1273
Falagas M, Betsi GI, Athanasiou S (2007) Probiotics for the treatment of women with bacterial vaginosis. Clin Microbiol Infect 13(7):657–664. https://doi.org/10.1111/j.1469-0691.2007.01688.x
Farisa Banu S, Rubini D, Murugan R, Vadivel V, Gowrishankar S, Pandian SK, Nithyanand P (2018a) Exploring the antivirulent and sea food preservation efficacy of essential oil combined with DNase on Vibrio parahaemolyticus. Lwt. https://doi.org/10.1016/j.lwt.2018.04.070
Farisa Banu S, Rubini D, Shanmugavelan P et al (2018b) Effects of patchouli and cinnamon essential oils on biofilm and hyphae formation by Candida species. J Mycol Med 28(2):332–339. https://doi.org/10.1016/j.mycmed.2018.02.012
Ferro TA, Araújo JM, Dos Santos Pinto BL et al (2016) Cinnamaldehyde inhibits staphylococcus aureus virulence factors and protects against infection in a galleria mellonella model. Front Microbiol 7:2052. https://doi.org/10.3389/fmicb.2016.02052
Ghannoum MA, Rice LB (1999) Antifungal agents: mode of action, mechanisms of resistance, and correlation of these mechanisms with bacterial resistance. Clin Microbiol Rev 12(4):501–517. https://doi.org/10.1128/CMR.12.4.501
Gulati M, Nobile CJ (2016) Candida albicans biofilms: development, regulation, and molecular mechanisms. Microbes Infect 18(5):310–321. https://doi.org/10.1016/j.micinf.2016.01.002
Hassan Y, Chew SY, Than LTL (2021) Candida glabrata: pathogenicity and resistance mechanisms for adaptation and survival. J Fungi (basel, Switzerland) 7(8):667. https://doi.org/10.3390/jof7080667
Hooper LV, Gordon JI (2001) Commensal host-bacterial relationships in the gut. Science 292(5519):1115–1118. https://doi.org/10.1126/science.1058709
Jayasankari S, Lakshmi D, Ramyadevi D, Vedha Hari BN, Anbazhagan V, Nithyanand P (2022) Evaluation of lectin nano scaffold based in-situ gel against vulvovaginal candidiasis causing Candida biofilms using a novel ex-vivo model. J Drug Deliv Sci Technol. https://doi.org/10.1016/j.jddst.2022.103560
Jeffery-Smith A, Taori SK, Schelenz S, Jeffery K, Johnson EM, Borman A, Manuel R, Brown CS, Candida auris Incident Management Team (2017) Candida auris: a review of the literature. Clin Microbiol Rev 31(1):e00029-e117. https://doi.org/10.1128/CMR.00029-17
Jothipandiyan S, Suresh D, Sankaran SV, Thamotharan S, Shanmugasundaram K, Vincent P, Sekaran S, Gowrishankar S, Pandian SK, Paramasivam N (2021) Heteroleptic pincer palladium(II) complex coated orthopedic implants impede the AbaI/ AbaR quorum sensing system and biofilm development by Acinetobacter baumannii. Biofouling. https://doi.org/10.1080/08927014.2021.2015336
Kim H, Kang SS (2019) Antifungal activities against Candida albicans, of cell-free supernatants obtained from probiotic Pediococcus acidilactici HW01. Arch Oral Biol 99:113–119. https://doi.org/10.1016/j.archoralbio.2019.01.006
Krishnasamy L, Rubini D, Senthilganesh J, Saikumar C, Kumaramanickavel G, Aruni AW, Nithyanand P (2020) Phylogenetic characterization of biofilm forming multidrug resistant Candida albicans and Non albicans Candida causing vulvovaginal candidiasis. Gene Reports 19:100644. https://doi.org/10.1016/j.genrep.2020.100644
Kuhn DM, Chandra J, Mukherjee PK, Ghannoum MA (2002) Comparison of biofilms formed by Candida albicans and Candida parapsilosis on bioprosthetic surfaces. Infect Immun 70(2):878–888. https://doi.org/10.1128/IAI.70.2.878-888.2002
Liong MT (2007) Probiotics: a critical review of their potential role as antihypertensives, immune modulators, hypocholesterolemics, and perimenopausal treatments. Nutr Rev 65(7):316–328. https://doi.org/10.1111/j.1753-4887.2007.tb00309.x
Markowiak P, Śliżewska K (2017) Effects of probiotics, prebiotics, and synbiotics on human health. Nutrients 9(9):1021. https://doi.org/10.3390/nu9091021
Mayer FL, Wilson D, Hube B (2013) Candida albicans pathogenicity mechanisms. Virulence 4(2):119–128. https://doi.org/10.4161/viru.22913
Mbakwem-Aniebo C, Osadebe AU, Athanasonny E, Okonko IO (2020) Prevalence of Candida spp. and age-related disparities amongst women presenting with vaginitis at the Obstetrics and Gynaecology (O&G) Clinic in a Tertiary hospital in Port Harcourt, Nigeria. Afr Health Sci 20(1):51–58. https://doi.org/10.4314/ahs.v20i1.9
Meng L, Zhao H, Zhao S, Sun X, Zhang M, Deng Y (2019) Inhibition of yeast-to-hypha transition and virulence of Candida albicans by 2-alkylaminoquinoline derivatives. Antimicrob Agents Chemother 63(4):e01891-e1918. https://doi.org/10.1128/AAC.01891-18
Muthamil S, Balasubramaniam B, Balamurugan K, Pandian SK (2018) Synergistic effect of quinic acid derived from syzygium cumini and undecanoic acid against candida spp. Biofilm and Virulence. Front Microbiol 9:2835. https://doi.org/10.3389/fmicb.2018.02835
Niimi M, Firth NA, Cannon RD (2010) Antifungal drug resistance of oral fungi. Odontology 98(1):15–25. https://doi.org/10.1007/s10266-009-0118-3
Nithyanand P, Beema Shafreen RM, Muthamil S, Karutha PS (2015) Usnic acid inhibits biofilm formation and virulent morphological traits of Candida albicans. Microbiol Res 179:20–28. https://doi.org/10.1016/j.micres.2015.06.009
Orsi CF, Sabia C, Ardizzoni A, Colombari B, Neglia RG, Peppoloni S et al (2014) Inhibitory effects of different lactobacilli on Candida albicans hyphal formation and biofilm development. J Biol Regul Homeost Agents 28:743–752
Parolin C, Croatti V, Laghi L et al (2021) Lactobacillus biofilms influence anti-candida activity. Front Microbiol 12:750368. https://doi.org/10.3389/fmicb.2021.750368
Raesi Vanani A, Mahdavinia M, Kalantari H, Khoshnood S, Shirani M (2019) Antifungal effect of the effect of Securigera securidaca L. vaginal gel on Candida species. Curr Med Mycol 5(3):31–35. https://doi.org/10.18502/cmm.5.3.1744
Reid G, Beuerman D, Heinemann C, Bruce AW (2001) Probiotic Lactobacillus dose required to restore and maintain a normal vaginal flora. FEMS Immunol Med Microbiol 32(1):37–41. https://doi.org/10.1111/j.1574-695X.2001.tb00531.x
Reid G, Jass J, Sebulsky MT, McCormick JK (2003) Potential uses of probiotics in clinical practice. Clin Microbiol Rev 4:658–672
Reygaert WC (2018) An overview of the antimicrobial resistance mechanisms of bacteria. AIMS Microbiol 4(3):482–501. https://doi.org/10.3934/microbiol.2018.3.482
Ribeiro FC, de Barros PP, Rossoni RD, Junqueira JC, Jorge AO (2017) Lactobacillus rhamnosus inhibits Candida albicans virulence factors in vitro and modulates the immune system in Galleria mellonella. J Appl Microbiol 122:201–211
Rolfe RD (2000) The role of probiotic cultures in the control of gastrointestinal health. J Nutr 130(2S Suppl):396S-402S. https://doi.org/10.1093/jn/130.2.396S
Rubini D, Farisa Banu S, Veda Hari BN et al (2018) Chitosan extracted from marine biowaste mitigates staphyloxanthin production and biofilms of Methicillin- resistant Staphylococcus aureus. Food Chem Toxicol 118:733–744. https://doi.org/10.1016/j.fct.2018.06.017
Rubini D, Vedha Hari BN, Nithyanand P (2021) Chitosan coated catheters alleviates mixed species biofilms ofStaphylococcus epidermidis and Candida albicans. Carbohydr Polym 252:117192 https://doi.org/10.1016/j.carbpol.2020.117192
Senthilganesh J, Kuppusamy S, Durairajan R, Subramaniyan SB, Veerappan A, Paramasivam N (2022) Phytolectin nanoconjugates in combination with standard antifungals curb multi-species biofilms and virulence of vulvovaginal candidiasis (VVC) causing Candida albicans and non-albicans Candida. Med Mycol 60(2):myab083. https://doi.org/10.1093/mmy/myab083
Shafreen RM, Muthamil S, Pandian SK (2014) Inhibition of Candida albicans virulence factors by novel levofloxacin derivatives. Appl Microbiol Biotechnol 98(15):6775–6785. https://doi.org/10.1007/s00253-014-5719-2
Silva S, Negri M, Henriques M, Oliveira R, Williams DW, Azeredo J (2012) Candida glabrata, Candida parapsilosis and Candida tropicalis: biology, epidemiology, pathogenicity and antifungal resistance. FEMS Microbiol Rev 36(2):288–305. https://doi.org/10.1111/j.1574-6976.2011.00278.x
Silva S, Rodrigues CF, Araújo D, Rodrigues ME, Henriques M (2017) Candida species biofilms’ antifungal resistance. J Fungi (basel) 3(1):8. https://doi.org/10.3390/jof3010008
Singh S, Singh SK, Chowdhury I, Singh R (2017) Understanding the mechanism of bacterial biofilms resistance to antimicrobial agents. Open Microbiol J 11:53–62. https://doi.org/10.2174/1874285801711010053
Spampinato C, Leonardi D (2013) Candida infections, causes, targets, and resistance mechanisms: traditional and alternative antifungal agents. Biomed Res Int 2013:204237. https://doi.org/10.1155/2013/204237
Srikanth R, Banu SF, Sowndarya J et al (2021) Biosurfactant synergized with marine bacterial DNase disrupts polymicrobial biofilms. Folia Microbiol (praha) 66(5):831–842. https://doi.org/10.1007/s12223-021-00876-y
Teodoro GR, Ellepola K, Seneviratne CJ, Koga-Ito CY (2015) Potential use of phenolic acids as anti-candida agents: a review. Front Microbiol 6:1420. https://doi.org/10.3389/fmicb.2015.01420
Vedha Hari BN, Narayanan N, Dhevedaran K (2015) Efavirenz–eudragit E-100 nanoparticle loaded aerosol foam for sustained release: in-vitro and ex-vivo evaluation. Chem Pap 69:358–368
Verdenelli MC, Coman MM, Cecchini C, Silvi S, Orpianesi C, Cresci A (2014) Evaluation of antipathogenic activity and adherence properties of human Lactobacillus strains for vaginal formulations. J Appl Microbiol 116(5):1297–1307. https://doi.org/10.1111/jam.12459
Wall G, Lopez-Ribot JL (2020) Current antimycotics, new prospects, and future approaches to antifungal therapy. Antibiotics (basel, Switzerland) 9(8):445. https://doi.org/10.3390/antibiotics9080445
Whaley SG, Berkow EL, Rybak JM, Nishimoto AT, Barker KS, Rogers PD (2017) Azole antifungal resistance in candida albicans and emerging non-albicans Candida species. Front Microbiol 7:2173. https://doi.org/10.3389/fmicb.2016.02173
Żółkiewicz J, Marzec A, Ruszczyński M, Feleszko W (2020) Postbiotics-a step beyond pre- and probiotics. Nutrients 12(8):2189. https://doi.org/10.3390/nu12082189
Acknowledgements
Financial support from the Indian Council of Medical Research (ICMR), New Delhi (OMI/18/2020-ECD-I) is gratefully acknowledged. The Fellowship to SJ in the form of JRF by ICMR is thankfully acknowledged.
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AS Methodology, Validation, Format Analysis, Writing- original draft, Visualization. KD Methodology, Validation, Format Analysis, Writing- original draft, Visualization. JS Methodology, Validation. DRD Review and Editing. BNVH Review and Editing.PN Conceptualization, Methodology, Validation, Format Analysis, Supervision, Funding Acquisition, Writing- Review & Editing.
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Kumar, S.A.S., Krishnan, D., Jothipandiyan, S. et al. Cell-free supernatants of probiotic consortia impede hyphal formation and disperse biofilms of vulvovaginal candidiasis causing Candida in an ex-vivo model. Antonie van Leeuwenhoek 117, 37 (2024). https://doi.org/10.1007/s10482-024-01929-1
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DOI: https://doi.org/10.1007/s10482-024-01929-1