Abstract
Candida albicans is the most common human fungal pathogen and can grow as yeast or filaments, depending on the environmental conditions. The filamentous form is of particular interest because it can play a direct role in adherence and pathogenicity. Therefore, the purpose of this study was to evaluate the effects of three clinical strains of Lactobacillus on C. albicans filamentation as well as their probiotic potential in pathogen-host interactions via an experimental candidiasis model study in Galleria mellonella. We used the reference strain Candida albicans ATCC 18804 and three clinical strains of Lactobacillus: L. rhamnosus strain 5.2, L. paracasei strain 20.3, and L. fermentum strain 20.4. First, the capacity of C. albicans to form hyphae was tested in vitro through association with the Lactobacillus strains. After that, we verified the ability of these strains to attenuate experimental candidiasis in a Galleria mellonella model through a survival curve assay. Regarding the filamentation assay, a significant reduction in hyphae formation of up to 57% was observed when C. albicans was incubated in the presence of the Lactobacillus strains, compared to a control group composed of only C. albicans. In addition, when the larvae were pretreated with Lactobacillus spp. prior to C. albicans infection, the survival rate of G. mellonela increased in all experimental groups. We concluded that Lactobacillus influences the growth and expression C. albicans virulence factors, which may interfere with the pathogenicity of these microorganisms.
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References
Alok A, Singh ID, Singh S, Kishore M, Jha PC, Iqubal MA (2017) Probiotics: a new era of biotherapy. Adv Biomed Res 6(1):31. https://doi.org/10.4103/2277-9175.192625
Badet C, Thebaud NB (2008) Ecology of lactobacilli in the oral cavity: a review of literature. Open Microbiol J 2(1):38–48. https://doi.org/10.2174/1874285800802010038
Bandara HM, Matsubara VH, Samaranayake LP (2017) Future therapies targeted towards eliminating Candida biofilms and associated infections. Expert Rev Anti-Infect Ther 15(3):299–318. https://doi.org/10.1080/14787210.2017.1268530
Bergin D, Brennan M, Kavanagh K (2003) Fluctuations in haemocyte density and microbial load may be used as indicators of fungal pathogenicity in larvae of Galleria mellonella. Microbes Infect 5(15):1389–1395. https://doi.org/10.1016/j.micinf.2003.09.019
Bergin D, Murphy L, Keenan J, Clynes M, Kavanagh K (2006) Pre-exposure to yeast protects larvae of Galleria mellonella from a subsequent lethal infection by Candida albicans and is mediated by the increased expression of antimicrobial peptides. Microbes Infect 8(8):2105–2112. https://doi.org/10.1016/j.micinf.2006.03.005
Borghi E, Romagnoli S, Fuchs BB, Cirasola D, Perdoni F, Tosi D, Braidotti P, Bulfamante G, Morace G, Mylonakis E (2014) Correlation between Candida albicans biofilm formation and invasion of the invertebrate host Galleria mellonella. Future Microbiol 9(2):163–173. https://doi.org/10.2217/fmb.13.159
Borghi E, Borgo F, Morace G (2016) Fungal biofilms: update on resistance. Adv Exp Med Biol 931:37–47. https://doi.org/10.1007/5584_2016_7
Coman MM, Verdenelli MC, Cecchini C, Silvi S, Orpianesi C, Boyko N, Cresci A (2014) In vitro evaluation of antimicrobial activity of Lactobacillus rhamnosus IMC 501((R)), Lactobacillus paracasei IMC 502((R)) and SYNBIO((R)) against pathogens. J Appl Microbiol 117(2):518–527. https://doi.org/10.1111/jam.12544
de Barros PP, Freire F, Rossoni RD, Junqueira JC, Jorge AOC (2017) Candida krusei and Candida glabrata reduce the filamentation of Candida albicans by downregulating expression of HWP1 gene. Folia Microbiol (Praha) 62(4):317–323. https://doi.org/10.1007/s12223-017-0500-4
de Oliveira FE, Rossoni RD, de Barros PP, Begnini BE, Junqueira JC, Jorge AOC, Leão MVP, de Oliveira LD (2017) Immunomodulatory effects and anti-Candida activity of lactobacilli in macrophages and in invertebrate model of Galleria mellonella. Microb Pathog 110:603–611. https://doi.org/10.1016/j.micpath.2017.08.006
Dubovskiy IM, Whitten MMA, Yaroslavtseva ON, Greig C, Kryukov VY, Grizanova EV, Mukherjee K, Vilcinskas A, Glupov VV, Butt TM (2013) Can insects develop resistance to insect pathogenic fungi? PLoS One 8(4):e60248. https://doi.org/10.1371/journal.pone.0060248
Egbe NE, Dornelles TO, Paget CM, Castelli LM, Ashe MP (2017) Farnesol inhibits translation to limit growth and filamentation in C. albicans and S. cerevisiae. Microb Cell 4(9):294–304. 10.15698/mic2017.09.589
Fedhila S, Buisson C, Dussurget O, Serror P, Glomski IJ, Liehl P, Lereclus D, Nielsen-LeRoux C (2010) Comparative analysis of the virulence of invertebrate and mammalian pathogenic bacteria in the oral insect infection model Galleria mellonella. J Invertebr Pathol 103(1):24–29. https://doi.org/10.1016/j.jip.2009.09.005
Fuchs BB, Eby J, Nobile CJ, El Khoury JB, Mitchell AP, Mylonakis E (2010) Role of filamentation in Galleria mellonella killing by Candida albicans. Microbes Infect 12(6):488–496. https://doi.org/10.1016/j.micinf.2010.03.001
Hashemi A, Villa CR, Comelli EM (2016) Probiotics in early life: a preventative and treatment approach. Food Funct 7(4):1752–1768. https://doi.org/10.1039/c5fo01148e
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(1):18. https://doi.org/10.1186/1472-6831-10-18
Höfs S, Mogavero S, Hube B (2016) Interaction of Candida albicans with host cells: virulence factors, host defense, escape strategies, and the microbiota. J Microbiol 54(3):149–169. https://doi.org/10.1007/s12275-016-5514-0
Junqueira JC (2012) Galleria mellonella as a model host for human pathogens: recent studies and new perspectives. Virulence 3(6):474–476. https://doi.org/10.4161/viru.22493
Kadosh D (2016) Control of Candida albicans morphology and pathogenicity by post-transcriptional mechanisms. Cell Mol Life Sci 73(22):4265–4278. https://doi.org/10.1007/s00018-016-2294-y
Leao MV, Goncalves Silva CR, Santos SS, Leite PG (2015) Lactobacillus rhamnosus may change the virulence of Candida albicans. Rev Bras Ginecol Obstet 37:417–420. https://doi.org/10.1590/SO100-720320150005217
Liaskovskii TM, Podgorskii VS (2005) Assessment of probiotics according to the international organizations (FAO/WHO). Mikrobiol Z 67(6):104–112
Matsubara VH, Bandara HM, Ishikawa KH, Mayer MP, Samaranayake LP (2016a) The role of probiotic bacteria in managing periodontal disease: a systematic review. Expert Rev Anti-Infect Ther 14(7):643–655. https://doi.org/10.1080/14787210.2016.1194198
Matsubara VH, Bandara HM, Mayer MP, Samaranayake LP (2016b) Probiotics as antifungals in mucosal candidiasis. Clin Infect Dis 62(9):1143–1153. https://doi.org/10.1093/cid/ciw038
Matsubara VH, Wang Y, Bandara HM, Mayer MP, Samaranayake LP (2016c) Probiotic lactobacilli inhibit early stages of Candida albicans biofilm development by reducing their growth, cell adhesion, and filamentation. Appl Microbiol Biotechnol 100(14):6415–6426. https://doi.org/10.1007/s00253-016-7527-3
Mayer FL, Wilson D, Hube B (2013) Candida albicans pathogenicity mechanisms. Virulence 4(2):119–128. https://doi.org/10.4161/viru.22913
Mc Namara L, Carolan JC, Griffin CT, Fitzpatrick D, Kavanagh K (2017) The effect of entomopathogenic fungal culture filtrate on the immune response of the greater wax moth, Galleria mellonella. J Insect Physiol 100:82–92. https://doi.org/10.1016/j.jinsphys.2017.05.009
Parahitiyawa NB et al (2006) Interspecies variation in Candida biofilm formation studied using the Calgary biofilm device. APMIS 114(4):298–306. https://doi.org/10.1111/j.1600-0463.2006.apm_394.x
Patel R, DuPont HL (2015) New approaches for bacteriotherapy: prebiotics, new-generation probiotics, and synbiotics. Clin Infect Dis 60(Suppl 2):S108–S121. https://doi.org/10.1093/cid/civ177
Peleg AY, Hogan DA, Mylonakis E (2010) Medically important bacterial-fungal interactions. Nat Rev Microbiol 8(5):340–349. https://doi.org/10.1038/nrmicro2313
Pujia AM, Costacurta M, Fortunato L, Merra G, Cascapera S, Calvani M, Gratteri S (2017) The probiotics in dentistry: a narrative review. Eur Rev Med Pharmacol Sci 21(6):1405–1412
Ribeiro FC, de Barros PP, Rossoni RD, Junqueira JC, Jorge AO (2017) Lactobacillus rhamnosus inhibits Candida albicans virulence factors in vitro and modulates immune system in Galleria mellonella. J Appl Microbiol 122(1):201–211. https://doi.org/10.1111/jam.13324
Rivera-Espinoza Y, Gallardo-Navarro Y (2010) Non-dairy probiotic products. Food Microbiol 27(1):1–11. https://doi.org/10.1016/j.fm.2008.06.008
Rossoni RD, Fuchs BB, de Barros PP, Velloso MD, Jorge AO, Junqueira JC, Mylonakis E (2017) Lactobacillus paracasei modulates the immune system of Galleria mellonella and protects against Candida albicans infection. PLoS One 12(3):e0173332. https://doi.org/10.1371/journal.pone.0173332
Smith AR, Macfarlane GT, Reynolds N, O'May GA, Bahrami B, Macfarlane S (2012) Effect of a synbiotic on microbial community structure in a continuous culture model of the gastric microbiota in enteral nutrition patients. FEMS Microbiol Ecol 80(1):135–145. https://doi.org/10.1111/j.1574-6941.2011.01279.x
Tati S, Davidow P, McCall A, Hwang-Wong E, Rojas IG, Cormack B, Edgerton M (2016) Candida glabrata binding to Candida albicans hyphae enables its development in oropharyngeal candidiasis. PLoS Pathog 12(3):e1005522. https://doi.org/10.1371/journal.ppat.1005522
Tsui C, Kong EF, Jabra-Rizk MA (2016) Pathogenesis of Candida albicans biofilm. Pathog Dis 74(4):ftw018. https://doi.org/10.1093/femspd/ftw018
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
Vila T, Romo JA, Pierce CG, McHardy SF, Saville SP, Lopez-Ribot JL (2017) Targeting Candida albicans filamentation for antifungal drug development. Virulence 8(2):150–158. https://doi.org/10.1080/21505594.2016.1197444
Vilela SF et al (2015) Lactobacillus acidophilus ATCC 4356 inhibits biofilm formation by C. albicans and attenuates the experimental candidiasis in Galleria mellonella. Virulence 6(1):29–39. https://doi.org/10.4161/21505594.2014.981486
Wu G, Xu L, Yi Y (2016) Galleria mellonella larvae are capable of sensing the extent of priming agent and mounting proportionatal cellular and humoral immune responses. Immunol Lett 174:45–52. https://doi.org/10.1016/j.imlet.2016.04.013
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This study was supported by the São Paulo Council of Research—FAPESP, Brazil (Grants 2013/25181-8 and 2015/09770-9).
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This study has been approved by the Ethics Committee of São Paulo State University (Unesp) under protocol 560.479.
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Rossoni, R.D., dos Santos Velloso, M., Figueiredo, L.M. et al. Clinical strains of Lactobacillus reduce the filamentation of Candida albicans and protect Galleria mellonella against experimental candidiasis. Folia Microbiol 63, 307–314 (2018). https://doi.org/10.1007/s12223-017-0569-9
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DOI: https://doi.org/10.1007/s12223-017-0569-9