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Applied Microbiology and Biotechnology

, Volume 100, Issue 14, pp 6415–6426 | Cite as

Probiotic lactobacilli inhibit early stages of Candida albicans biofilm development by reducing their growth, cell adhesion, and filamentation

  • Victor Haruo Matsubara
  • Yi Wang
  • H. M. H. N. Bandara
  • Marcia Pinto Alves Mayer
  • Lakshman P. SamaranayakeEmail author
Applied microbial and cell physiology

Abstract

We evaluated the inhibitory effects of the probiotic Lactobacillus species on different phases of Candida albicans biofilm development. Quantification of biofilm growth and ultrastructural analyses were performed on C. albicans biofilms treated with Lactobacillus rhamnosus, Lactobacillus casei, and Lactobacillus acidophilus planktonic cell suspensions as well as their supernatants. Planktonic lactobacilli induced a significant reduction (p < 0.05) in the number of biofilm cells (25.5–61.8 %) depending on the probiotic strain and the biofilm phase. L. rhamnosus supernatants had no significant effect on the mature biofilm (p > 0.05), but significantly reduced the early stages of Candida biofilm formation (p < 0.01). Microscopic analyses revealed that L. rhamnosus suspensions reduced Candida hyphal differentiation, leading to a predominance of budding growth. All lactobacilli negatively impacted C. albicans yeast-to-hyphae differentiation and biofilm formation. The inhibitory effects of the probiotic Lactobacillus on C. albicans entailed both cell-cell interactions and secretion of exometabolites that may impact on pathogenic attributes associated with C. albicans colonization on host surfaces and yeast filamentation. This study clarifies, for the first time, the mechanics of how Lactobacillus species may antagonize C. albicans host colonization. Our data elucidate the inhibitory mechanisms that define the probiotic candicidal activity of lactobacilli, thus supporting their utility as an adjunctive therapeutic mode against mucosal candidal infections.

Keywords

Biofilm Candida albicans Candidiasis Lactobacillus Probiotics 

Notes

Acknowledgments

The authors thank Carol Tran for the technical support with the scanning electron microscope and the Coordination for the Improvement of Higher Education Personnel (CAPES) Foundation for supporting Victor H. Matsubara.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

References

  1. Alcazar-Fuoli L, Mellado E (2014) Current status of antifungal resistance and its impact on clinical practice. Br J Haematol 166(4):471–484. doi: 10.1111/bjh.12896 CrossRefPubMedGoogle Scholar
  2. Anderson JM, Soll DR (1986) Differences in actin localization during bud and hypha formation in the yeast Candida albicans. J Gen Microbiol 132(7):2035–2047. doi: 10.1099/00221287-132-7-2035 PubMedGoogle Scholar
  3. Arendrup MC (2010) Epidemiology of invasive candidiasis. Curr Opin Crit Care 16(5):445–452. doi: 10.1097/MCC.0b013e32833e84d2 CrossRefPubMedGoogle Scholar
  4. Atanassova M, Choiset Y, Dalgalarrondo M, Chobert JM, Dousset X, Ivanova I, Haertle T (2003) Isolation and partial biochemical characterization of a proteinaceous anti-bacteria and anti-yeast compound produced by Lactobacillus paracasei subsp. paracasei strain M3. Int J Food Microbiol 87(1–2):63–73. doi: 10.1016/S0168-1605(03)00054-0 CrossRefPubMedGoogle Scholar
  5. Bandara HM, Cheung BP, Watt RM, Jin LJ, Samaranayake LP (2013) Secretory products of Escherichia coli biofilm modulate Candida biofilm formation and hyphal development. J Investig Clin Dent 4(3):186–199. doi: 10.1111/jicd.12048 CrossRefPubMedGoogle Scholar
  6. Ceresa C, Tessarolo F, Caola I, Nollo G, Cavallo M, Rinaldi M, Fracchia L (2015) Inhibition of Candida albicans adhesion on medical-grade silicone by a Lactobacillus-derived biosurfactant. J Appl Microbiol 118(5):1116–1125. doi: 10.1111/jam.12760 CrossRefPubMedGoogle Scholar
  7. Chew SY, Cheah YK, Seow HF, Sandai D, Than LT (2015a) In vitro modulation of probiotic bacteria on the biofilm of Candida glabrata. Anaerobe 34:132–138. doi: 10.1016/j.anaerobe.2015.05.009 CrossRefPubMedGoogle Scholar
  8. Chew SY, Cheah YK, Seow HF, Sandai D, Than LT (2015b) Probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 exhibit strong antifungal effects against vulvovaginal candidiasis-causing Candida glabrata isolates. J Appl Microbiol 118:1180–1190. doi: 10.1111/jam.12772 CrossRefPubMedPubMedCentralGoogle Scholar
  9. Coleman JJ, Okoli I, Tegos GP, Holson EB, Wagner FF, Hamblin MR, Mylonakis E (2010) Characterization of plant-derived saponin natural products against Candida albicans. ACS Chem Biol 5(3):321–332. doi: 10.1021/cb900243b CrossRefPubMedPubMedCentralGoogle Scholar
  10. Ericson JE, Benjamin DK Jr (2014) Fluconazole prophylaxis for prevention of invasive candidiasis in infants. Curr Opin Pediatr 26(2):151–156. doi: 10.1097/MOP.0000000000000060 CrossRefPubMedPubMedCentralGoogle Scholar
  11. Falagas ME, Betsi GI, Athanasiou S (2006) Probiotics for prevention of recurrent vulvovaginal candidiasis: a review. J Antimicrob Chemother 58(2):266–272. doi: 10.1093/jac/dkl246 CrossRefPubMedGoogle Scholar
  12. Fidan I, Kalkanci A, Yesilyurt E, Yalcin B, Erdal B, Kustimur S, Imir T (2009) Effects of Saccharomyces boulardii on cytokine secretion from intraepithelial lymphocytes infected by Escherichia coli and Candida albicans. Mycoses 52(1):29–34. doi: 10.1111/j.1439-0507.2008.01545.x CrossRefPubMedGoogle Scholar
  13. Hasslof P, Hedberg M, Twetman S, Stecksen-Blicks C (2010) Growth inhibition of oral mutans streptococci and Candida by commercial probiotic lactobacilli—an in vitro study. BMC Oral Health 10:18. doi: 10.1186/1472-6831-10-18 CrossRefPubMedPubMedCentralGoogle Scholar
  14. Hatakka K, Ahola AJ, Yli-Knuuttila H, Richardson M, Poussa T, Meurman JH, Korpela R (2007) Probiotics reduce the prevalence of oral Candida in the elderly—a randomized controlled trial. J Dent Res 86(2):125–130CrossRefPubMedGoogle Scholar
  15. Hu H, Merenstein DJ, Wang C, Hamilton PR, Blackmon ML, Chen H, Calderone RA, Li D (2013) Impact of eating probiotic yogurt on colonization by Candida species of the oral and vaginal mucosa in HIV-infected and HIV-uninfected women. Mycopathologia 176(3–4):175–181. doi: 10.1007/s11046-013-9678-4 CrossRefPubMedPubMedCentralGoogle Scholar
  16. Ishikawa KH, Mayer MP, Miyazima TY, Matsubara VH, Silva EG, Paula CR, Campos TT, Nakamae AE (2014) A multispecies probiotic reduces oral Candida colonization in denture wearers. J Prosthodont 24(3):194–199. doi: 10.1111/jopr.12198 CrossRefPubMedGoogle Scholar
  17. Jin Y, Zhang T, Samaranayake YH, Fang HH, Yip HK, Samaranayake LP (2005) The use of new probes and stains for improved assessment of cell viability and extracellular polymeric substances in Candida albicans biofilms. Mycopathologia 159(3):353–360. doi: 10.1007/s11046-004-6987-7 CrossRefPubMedGoogle Scholar
  18. Jones T, Federspiel NA, Chibana H, Dungan J, Kalman S, Magee BB, Newport G, Thorstenson YR, Agabian N, Magee PT, Davis RW, Scherer S (2004) The diploid genome sequence of Candida albicans. Proc Natl Acad Sci U S A 101(19):7329–7334. doi: 10.1073/pnas.0401648101 CrossRefPubMedPubMedCentralGoogle Scholar
  19. Kheradmand E, Rafii F, Yazdi MH, Sepahi AA, Shahverdi AR, Oveisi MR (2014) The antimicrobial effects of selenium nanoparticle-enriched probiotics and their fermented broth against Candida albicans. Daru 22:48. doi: 10.1186/2008-2231-22-48 CrossRefPubMedPubMedCentralGoogle Scholar
  20. Kohler GA, Assefa S, Reid G (2012) Probiotic interference of Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 with the opportunistic fungal pathogen Candida albicans. Infect Dis Obstet Gynecol 2012:636474. doi: 10.1155/2012/636474 CrossRefPubMedPubMedCentralGoogle Scholar
  21. Kovachev SM, Vatcheva-Dobrevska RS (2015) Local probiotic therapy for vaginal Candida albicans infections. Probiotics Antimicrob Proteins 7(1):38–44. doi: 10.1007/s12602-014-9176-0 CrossRefPubMedGoogle Scholar
  22. Kraft-Bodi E, Jorgensen MR, Keller MK, Kragelund C, Twetman S (2015) Effect of probiotic bacteria on oral Candida in frail elderly. J Dent Res 94(9):181–186. doi: 10.1177/0022034515595950 CrossRefGoogle Scholar
  23. Kucharikova S, Tournu H, Lagrou K, Van Dijck P, Bujdakova H (2011) Detailed comparison of Candida albicans and Candida glabrata biofilms under different conditions and their susceptibility to caspofungin and anidulafungin. J Med Microbiol 60:1261–1269. doi: 10.1099/jmm.0.032037-0 CrossRefPubMedGoogle Scholar
  24. Li D, Li Q, Liu C, Lin M, Li X, Xiao X, Zhu Z, Gong Q, Zhou H (2014) Efficacy and safety of probiotics in the treatment of Candida-associated stomatitis. Mycoses 57(3):141–146. doi: 10.1111/myc.12116 CrossRefPubMedGoogle Scholar
  25. Li H, Liu L, Zhang S, Cui W, Lv J (2012) Identification of antifungal compounds produced by Lactobacillus casei AST18. Curr Microbiol 65(2):156–161. doi: 10.1007/s00284-012-0135-2 CrossRefPubMedGoogle Scholar
  26. Maldonado-Barragan A, Caballero-Guerrero B, Lucena-Padros H, Ruiz-Barba JL (2013) Induction of bacteriocin production by coculture is widespread among plantaricin-producing Lactobacillus plantarum strains with different regulatory operons. Food Microbiol 33(1):40–47. doi: 10.1016/j.fm.2012.08.009 CrossRefPubMedGoogle Scholar
  27. Matsubara VH, Bandara HM, Mayer MP, Samaranayake LP (2016) Probiotics as antifungals in mucosal candidiasis. Clin Infect Dis. doi: 10.1093/cid/ciw038 PubMedGoogle Scholar
  28. Matsubara VH, Silva EG, Paula CR, Ishikawa KH, Nakamae AE (2012) Treatment with probiotics in experimental oral colonization by Candida albicans in murine model (DBA/2). Oral Dis 18(3):260–264. doi: 10.1111/j.1601-0825.2011.01868.x CrossRefPubMedGoogle Scholar
  29. Mendonça FH, Santos SS, Faria Ida S, Silva CRG e, AO J, MV L (2012) Effects of probiotic bacteria on Candida presence and IgA anti-Candida in the oral cavity of elderly. Braz Dent J 23(5):534–538. doi: 10.1590/S0103-64402012000500011 CrossRefPubMedGoogle Scholar
  30. Meurman JH (2005) Probiotics: do they have a role in oral medicine and dentistry? Eur J Oral Sci 113(3):188–196CrossRefPubMedGoogle Scholar
  31. Miller MB, Bassler BL (2001) Quorum sensing in bacteria. Annu Rev Microbiol 55:165–199. doi: 10.1146/annurev.micro.55.1.165 CrossRefPubMedGoogle Scholar
  32. Murzyn A, Krasowska A, Stefanowicz P, Dziadkowiec D, Lukaszewicz M (2010) Capric acid secreted by S. boulardii inhibits C. albicans filamentous growth, adhesion and biofilm formation. PLoS One 5(8):e12050. doi: 10.1371/journal.pone.0012050 CrossRefPubMedPubMedCentralGoogle Scholar
  33. Nett JE (2014) Future directions for anti-biofilm therapeutics targeting Candida. Expert Rev Anti-Infect Ther 12(3):375–382. doi: 10.1586/14787210.2014.885838 CrossRefPubMedGoogle Scholar
  34. Orsi CF, Sabia C, Ardizzoni A, Colombari B, Neglia RG, Peppoloni S, Morace G, Blasi E (2014) Inhibitory effects of different lactobacilli on Candida albicans hyphal formation and biofilm development. J Biol Regul Homeost Agents 28(4):743–752PubMedGoogle Scholar
  35. Pappas PG (2014) Antifungal clinical trials and guidelines: what we know and do not know. Cold Spring Harb Perspect Med 4(11):a019745. doi: 10.1101/cshperspect.a019745 CrossRefPubMedGoogle Scholar
  36. Parolin C, Marangoni A, Laghi L, Foschi C, Nahui Palomino RA, Calonghi N, Cevenini R, Vitali B (2015) Isolation of vaginal lactobacilli and characterization of anti-Candida activity. PLoS One 10(6):e0131220. doi: 10.1371/journal.pone.0131220 CrossRefPubMedPubMedCentralGoogle Scholar
  37. Polke M, Hube B, Jacobsen ID (2015) Candida survival strategies. Adv Appl Microbiol 91:139–235. doi: 10.1016/bs.aambs.2014.12.002 CrossRefPubMedGoogle Scholar
  38. Rizzello CG, Filannino P, Di Cagno R, Calasso M, Gobbetti M (2014) Quorum-sensing regulation of constitutive plantaricin by Lactobacillus plantarum strains under a model system for vegetables and fruits. Appl Environ Microbiol 80(2):777–787. doi: 10.1128/AEM.03224-13 CrossRefPubMedPubMedCentralGoogle Scholar
  39. Rodrigues L, Banat IM, Teixeira J, Oliveira R (2006a) Biosurfactants: potential applications in medicine. J Antimicrob Chemother 57(4):609–618. doi: 10.1093/jac/dkl024 CrossRefPubMedGoogle Scholar
  40. Rodrigues LR, Teixeira JA, van der Mei HC, Oliveira R (2006b) Physicochemical and functional characterization of a biosurfactant produced by Lactococcus lactis 53. Colloids Surf B Biointerfaces 49(1):79–86. doi: 10.1016/j.colsurfb.2006.03.003 CrossRefPubMedGoogle Scholar
  41. Roy A, Chaudhuri J, Sarkar D, Ghosh P, Chakraborty S (2014) Role of enteric supplementation of probiotics on late-onset sepsis by Candida species in preterm low birth weight neonates: a randomized, double blind, placebo-controlled trial. N Am J Med Sci 6(1):50–57. doi: 10.4103/1947-2714.125870 CrossRefPubMedPubMedCentralGoogle Scholar
  42. Ryu EH, Yang EJ, Woo ER, Chang HC (2014) Purification and characterization of antifungal compounds from Lactobacillus plantarum HD1 isolated from kimchi. Food Microbiol 41:19–26. doi: 10.1016/j.fm.2014.01.011 CrossRefPubMedGoogle Scholar
  43. Samaranayake LP (2012) Essential microbiology for dentistry, 4th edn. Churchill Livingstone, EdinburghGoogle Scholar
  44. Samaranayake LP, Cheung LK, Samaranayake YH (2002) Candidiasis and other fungal diseases of the mouth. Dermatol Ther 15:252–270. doi: 10.1046/j.1529-8019.2002.01533.x CrossRefGoogle Scholar
  45. Sanguinetti M, Posteraro B, Lass-Florl C (2015) Antifungal drug resistance among Candida species: mechanisms and clinical impact. Mycoses 58(Suppl 2):2–13. doi: 10.1111/myc.12330 CrossRefPubMedGoogle Scholar
  46. Sardi JC, Scorzoni L, Bernardi T, Fusco-Almeida AM, Mendes Giannini MJ (2013) Candida species: current epidemiology, pathogenicity, biofilm formation, natural antifungal products and new therapeutic options. J Med Microbiol 62(Pt 1):10–24. doi: 10.1099/jmm.0.045054-0 CrossRefPubMedGoogle Scholar
  47. Servin AL, Coconnier MH (2003) Adhesion of probiotic strains to the intestinal mucosa and interaction with pathogens. Best Pract Res Clin Gastroenterol 17(5):741–754. doi: 10.1016/S1521-6918(03)00052-0 CrossRefPubMedGoogle Scholar
  48. Sherry L, Jose A, Murray C, Williams C, Jones B, Millington O, Bagg J, Ramage G (2012) Carbohydrate derived fulvic acid: an in vitro investigation of a novel membrane active antiseptic agent against Candida albicans biofilms. Front Microbiol 3:116. doi: 10.3389/fmicb.2012.00116 CrossRefPubMedPubMedCentralGoogle Scholar
  49. Simark-Mattsson C, Jonsson R, Emilson CG, Roos K (2009) Final pH affects the interference capacity of naturally occurring oral Lactobacillus strains against mutans streptococci. Arch Oral Biol 54(6):602–607. doi: 10.1016/j.archoralbio.2009.03.005 CrossRefPubMedGoogle Scholar
  50. Strus M, Kucharska A, Kukla G, Brzychczy-Wloch M, Maresz K, Heczko PB (2005) The in vitro activity of vaginal Lactobacillus with probiotic properties against Candida. Infect Dis Obstet Gynecol 13(2):69–75CrossRefPubMedPubMedCentralGoogle Scholar
  51. Taff HT, Mitchell KF, Edward JA, Andes DR (2013) Mechanisms of Candida biofilm drug resistance. Future Microbiol 8(10):1325–1337. doi: 10.2217/fmb.13.101 CrossRefPubMedGoogle Scholar
  52. 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. doi: 10.1111/jam.12459 CrossRefPubMedGoogle Scholar
  53. Vilela SF, Barbosa JO, Rossoni RD, Santos JD, Prata MC, Anbinder AL, Jorge AO, Junqueira JC (2015) Lactobacillus acidophilus ATCC 4356 inhibits biofilm formation by C. albicans and attenuates the experimental candidiasis in Galleria mellonella. Virulence 6(1):29–39. doi: 10.4161/21505594.2014.981486 CrossRefPubMedPubMedCentralGoogle Scholar
  54. Wang H, Yan Y, Wang J, Zhang H, Qi W (2012) Production and characterization of antifungal compounds produced by Lactobacillus plantarum IMAU10014. PLoS One 7(1):e29452. doi: 10.1371/journal.pone.0029452 CrossRefPubMedPubMedCentralGoogle Scholar
  55. Zakaria Gomaa E (2013) Antimicrobial and anti-adhesive properties of biosurfactant produced by lactobacilli isolates, biofilm formation and aggregation ability. J Gen Appl Microbiol 59(6):425–436. doi: 10.2323/jgam.59.425 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Victor Haruo Matsubara
    • 1
    • 2
    • 3
  • Yi Wang
    • 1
  • H. M. H. N. Bandara
    • 1
  • Marcia Pinto Alves Mayer
    • 2
  • Lakshman P. Samaranayake
    • 1
    Email author
  1. 1.UQ Oral Health Centre, School of DentistryThe University of QueenslandBrisbaneAustralia
  2. 2.Department of Microbiology, Institute of Biomedical SciencesUniversity of São PauloSão PauloBrazil
  3. 3.Department of Prosthodontics, School of DentistryUniversity of São PauloSão PauloBrazil

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