Evaluation of antifungal and antibiofilm effects of supernatant cell-free supernatant (CFS) of Lactobacillus casei alone and in combination with nystatin against Candida albicans strains
-
Omolbanin Faryabi
,
Elham Isaei
,
Samira Salari
,
Seyed Amin Ayatollahi Mousavi
,
Ehsan Salarkia
,
Mohammad Hossein Sobhanipoor
,
Pooya Ghasemi Nejad Almani
,
Alireza Izadi
Abstract
Background and Objectives: Candida albicans (C. albicans) is the most common cause of candidiasis in immunocompromised patients. Biofilm-associated Candida can lead to treatment complications and systemic infections.
Materials and Methods: In this study, the antifungal activity of the cell-free supernatant (CFS) derived from Lactobacillus casei was assessed against C. albicans strains. To differentiate between pH- and H₂O₂-mediated effects and specific metabolite activity, all antifungal and antibiofilm assays were performed using CFS that had been neutralized to pH 7.0 and treated with catalase.
Results: The ranges of minimum inhibitory concentration (MIC) and Half-maximal inhibitory concentration (IC50) for CFS were 7.81-62.5 and 95.1-159.6 µl/ml, respectively. Using corrected FICI criteria (synergy ≤ 0.5; additive > 0.5-1.0; indifference > 1.0-4.0), additive effects were observed in 50% of strains, synergy in 20% (including the standard strain PTCC 5027), and indifference in 30%.
Conclusion: The results demonstrate that the CFS of L. casei exhibits promising in vitro antifungal and antibiofilm activity against C. albicans. When used in combination with nystatin, L. casei CFS showed additive or synergistic effects, suggesting its potential as an adjunctive agent for further investigation. However, these findings are preliminary, and in vivo studies and clinical assessments are necessary to validate the therapeutic potential of L. casei CFS in the management of oral candidiasis.
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11. Wilkins T, Sequoia J. Probiotics for Gastrointestinal Conditions: A Summary of the Evidence. Am Fam Physician 2017; 96: 170-178.
12. Boirivant M, Strober W. The mechanism of action of probiotics. Curr Opin Gastroenterol 2007; 23: 679-692.
13. Davati N, Tabatabaee Yazdi F, Zibaee S, Shahidi F, Edalatian MR. Study of Lactic Acid Bacteria Community From Raw Milk of Iranian One Humped Camel and Evaluation of Their Probiotic Properties. Jundishapur J Microbiol 2015; 8(5): e16750.
14. Salari S, Sadat Seddighi N, Ghasemi Nejad Almani P. Evaluation of biofilm formation ability in different Candida strains and anti-biofilm effects of Fe3O4-NPs compared with Fluconazole: an in vitro study. J Mycol Med 2018; 28: 23-28.
15. Poon Y, Hui M. Inhibitory effect of lactobacilli supernatants on biofilm and filamentation of Candida albicans, Candida tropicalis, and Candida parapsilosis. Front Microbiol 2023; 14: 1105949.
16. Martins N, Ferreira IC, Barros L, Silva S, Henriques M. Candidiasis: predisposing factors, prevention, diagnosis and alternative treatment. Mycopathologia 2014; 177: 223-240.
17. Denning DW. Global incidence and mortality of severe fungal disease. Lancet Infect Dis 2024; 24(7): e428-e438.
18. Salari S, Ghasemi Nejad Almani P. Antifungal effects of Lactobacillus acidophilus and Lactobacillus plantarum against different oral Candida species isolated from HIV/ AIDS patients: an in vitro study. J Oral Microbiol 2020; 12: 1769386.
19. Jørgensen MR, Kragelund C, Jensen PØ, Keller MK, Twetman S. Probiotic Lactobacillus reuteri has antifungal effects on oral Candida species in vitro. J Oral Microbiol 2017; 9: 1274582.
20. Ang XY, Mageswaran UM, Chung YLF, Lee BK, Azhar SNA, Roslan NS, et al. Probiotics Reduce Vaginal Candidiasis in Pregnant Women via Modulating Abundance of Candida and Lactobacillus in Vaginal and Cervicovaginal Regions. Microorganisms 2022; 10: 285.
21. Biyari S, Fozouni L. The inhibitory effect of probiotic bacteria against drug resistant Candida species isolated from the oral cavity of the elderly. Shiraz E Med J 2018; 19(6): e62026.
22. Sharifzadeh SS, Lotfali E, Lesan S, Farrokhnia T. Antifungal Effect of Probiotic Lactobacillus casei on Drug-Resistant Oral Candida albicans Isolated from Patients with Hematological Malignancy: an in vitroStudy. J Dent (Shiraz) 2023; 24: 389-394.
23. Chew SY, Cheah YK, Seow HF, Sandai D, Than LT. Probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 exhibit strong antifungal effects against vulvovaginal candidiasis-causing Candida glabrata isolates. J Appl Microbiol 2015; 118: 1180-1190.
24. Ahire JJ, Sahoo S, Kashikar MS, Heerekar A, Lakshmi SG, Madempudi RS. In vitro assessment of Lactobacillus crispatus UBLCp01, Lactobacillus gasseri UBLG36, and Lactobacillus johnsonii UBLJ01 as a Potential Vaginal Probiotic Candidate. Probiotics Antimicro 2023; 15: 275-286.
25. Vylkova S. Environmental pH modulation by pathogenic fungi as a strategy to conquer the host. PLoS Pathog 2017; 13(2): e1006149.
26. Andrade JC, Kumar S, Kumar A, Černáková L, Rodrigues CF. Application of probiotics in candidiasis management. Crit Rev Food Sci Nutr 2022; 62: 8249-8264.
27. Pedro NA, Fontebasso G, Pinto SN, Alves M, Mira NP. Acetate modulates the inhibitory effect of Lactobacillus gasseri against the pathogenic yeasts Candida albicans and Candida glabrata. Microb Cell 2023; 10: 88-102.
28. Uppuluri P, Pierce CG, López-Ribot JL. Candida albicans biofilm formation and its clinical consequences. Future Microbiol 2009; 4: 1235-1237.
29. Hu L, Zhou M, Young A, Zhao W, Yan Z. In vivo effectiveness and safety of probiotics on prophylaxis and treatment of oral candidiasis: a systematic review and meta-analysis. BMC Oral Health 2019; 19: 140.
30. Srivastava N, Ellepola K, Venkiteswaran N, Chai LYA, Ohshima T, Seneviratne CJ. Lactobacillus Plantarum 108 Inhibits Streptococcus mutans and Candida albicans Mixed-Species Biofilm Formation. Antibiotics (Basel) 2020; 9: 478.
31. Ai R, Wei J, Ma D, Jiang L, Dan H, Zhou Y, et al. A meta-analysis of randomized trials assessing the effects of probiotic preparations on oral candidiasis in the elderly. Arch Oral Biol 2017; 83: 187-192.
32. Zeng Y, Fadaak A, Alomeir N, Wu Y, Wu TT, Qing S, et al. Effect of Probiotic Lactobacillus plantarum on Streptococcus mutans and Candida albicans Clinical Isolates from Children with Early Childhood Caries. Int J Mol Sci 2023; 24: 2991.
33. Rather IA, Wani MY, Kamli MR, Sabir JSM, Hakeem KR, Firoz A, et al. Limosilactobacillus fermentum KAU0021 Abrogates Mono- and Polymicrobial Biofilms Formed by Candida albicans and Staphylococcus aureus. Pharmaceutics 2023; 15: 1079.
34. Peeters E, Nelis HJ, Coenye T. Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates. J Microbiol Methods 2008; 72: 157-165.
35. Stepanović S, Vuković D, Hola V, Di Bonaventura G, Djukić S, Cirković I, et al. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS 2007; 115: 891-899.
36. Chandra J, Kuhn DM, Mukherjee PK, Hoyer LL, McCormick T, Ghannoum MA. Biofilm formation by the fungal pathogen Candida albicans: development, architecture, and drug resistance. J Bacteriol 2001; 183: 5385-5394.
37. Jakubovics NS, Goodman SD, Mashburn-Warren L, Stafford GP, Cieplik F. The dental plaque biofilm matrix. Periodontol 2000 2021; 86: 32-56.
2. Neppelenbroek KH, Seó RS, Urban VM, Silva S, Dovigo LN, Jorge JH, et al. Identification of Candida species in the clinical laboratory: a review of conventional, commercial, and molecular techniques. Oral Dis 2014; 20: 329-444.
3. Millsop JW, Fazel N. Oral candidiasis. Clin Dermatol 2016; 34: 487-494.
4. Jafarian H, Gharaghani M, Asnafi AA, Hardani AK, Zarei-Mahmoudabadi A. Phenotype, genotype, and mating type determination in oral Candida albicans isolates from pediatric patients with neutropenia. J Clin Lab Anal 2022; 36(9): e24664.
5. Cortegiani A, Misseri G, Fasciana T, Giammanco A, Giarratano A, Chowdhary A. Epidemiology, clinical characteristics, resistance, and treatment of infections by Candida auris. J Intensive Care 2018; 6: 69.
6. Bandara HM, Matsubara VH, Samaranayake LP. Future therapies targeted towards eliminating Candida biofilms and associated infections. Expert Rev Anti Infect Ther 2017; 15: 299-318.
7. Mazziotta C, Tognon M, Martini F, Torreggiani E, Rotondo JC. Probiotics Mechanism of Action on Immune Cells and Beneficial Effects on Human Health. Cells 2023; 12: 184.
8. Matsubara VH, Bandara HM, Mayer MP, Samaranayake LP. Probiotics as Antifungals in Mucosal Candidiasis. Clin Infect Dis 2016; 62: 1143-1153.
9. Ribeiro FC, Rossoni RD, de Barros PP, Santos JD, Fugisaki LRO, Leão MPV, et al. Action mechanisms of probiotics on Candida spp. and candidiasis prevention: an update. J Appl Microbiol 2020; 129: 175-185.
10. Parolin C, Marangoni A, Laghi L, Foschi C, Ñahui Palomino RA, Calonghi N, et al. Isolation of Vaginal Lactobacilli and Characterization of Anti-Candida Activity. PLoS One 2015; 10(6): e0131220.
11. Wilkins T, Sequoia J. Probiotics for Gastrointestinal Conditions: A Summary of the Evidence. Am Fam Physician 2017; 96: 170-178.
12. Boirivant M, Strober W. The mechanism of action of probiotics. Curr Opin Gastroenterol 2007; 23: 679-692.
13. Davati N, Tabatabaee Yazdi F, Zibaee S, Shahidi F, Edalatian MR. Study of Lactic Acid Bacteria Community From Raw Milk of Iranian One Humped Camel and Evaluation of Their Probiotic Properties. Jundishapur J Microbiol 2015; 8(5): e16750.
14. Salari S, Sadat Seddighi N, Ghasemi Nejad Almani P. Evaluation of biofilm formation ability in different Candida strains and anti-biofilm effects of Fe3O4-NPs compared with Fluconazole: an in vitro study. J Mycol Med 2018; 28: 23-28.
15. Poon Y, Hui M. Inhibitory effect of lactobacilli supernatants on biofilm and filamentation of Candida albicans, Candida tropicalis, and Candida parapsilosis. Front Microbiol 2023; 14: 1105949.
16. Martins N, Ferreira IC, Barros L, Silva S, Henriques M. Candidiasis: predisposing factors, prevention, diagnosis and alternative treatment. Mycopathologia 2014; 177: 223-240.
17. Denning DW. Global incidence and mortality of severe fungal disease. Lancet Infect Dis 2024; 24(7): e428-e438.
18. Salari S, Ghasemi Nejad Almani P. Antifungal effects of Lactobacillus acidophilus and Lactobacillus plantarum against different oral Candida species isolated from HIV/ AIDS patients: an in vitro study. J Oral Microbiol 2020; 12: 1769386.
19. Jørgensen MR, Kragelund C, Jensen PØ, Keller MK, Twetman S. Probiotic Lactobacillus reuteri has antifungal effects on oral Candida species in vitro. J Oral Microbiol 2017; 9: 1274582.
20. Ang XY, Mageswaran UM, Chung YLF, Lee BK, Azhar SNA, Roslan NS, et al. Probiotics Reduce Vaginal Candidiasis in Pregnant Women via Modulating Abundance of Candida and Lactobacillus in Vaginal and Cervicovaginal Regions. Microorganisms 2022; 10: 285.
21. Biyari S, Fozouni L. The inhibitory effect of probiotic bacteria against drug resistant Candida species isolated from the oral cavity of the elderly. Shiraz E Med J 2018; 19(6): e62026.
22. Sharifzadeh SS, Lotfali E, Lesan S, Farrokhnia T. Antifungal Effect of Probiotic Lactobacillus casei on Drug-Resistant Oral Candida albicans Isolated from Patients with Hematological Malignancy: an in vitroStudy. J Dent (Shiraz) 2023; 24: 389-394.
23. Chew SY, Cheah YK, Seow HF, Sandai D, Than LT. Probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 exhibit strong antifungal effects against vulvovaginal candidiasis-causing Candida glabrata isolates. J Appl Microbiol 2015; 118: 1180-1190.
24. Ahire JJ, Sahoo S, Kashikar MS, Heerekar A, Lakshmi SG, Madempudi RS. In vitro assessment of Lactobacillus crispatus UBLCp01, Lactobacillus gasseri UBLG36, and Lactobacillus johnsonii UBLJ01 as a Potential Vaginal Probiotic Candidate. Probiotics Antimicro 2023; 15: 275-286.
25. Vylkova S. Environmental pH modulation by pathogenic fungi as a strategy to conquer the host. PLoS Pathog 2017; 13(2): e1006149.
26. Andrade JC, Kumar S, Kumar A, Černáková L, Rodrigues CF. Application of probiotics in candidiasis management. Crit Rev Food Sci Nutr 2022; 62: 8249-8264.
27. Pedro NA, Fontebasso G, Pinto SN, Alves M, Mira NP. Acetate modulates the inhibitory effect of Lactobacillus gasseri against the pathogenic yeasts Candida albicans and Candida glabrata. Microb Cell 2023; 10: 88-102.
28. Uppuluri P, Pierce CG, López-Ribot JL. Candida albicans biofilm formation and its clinical consequences. Future Microbiol 2009; 4: 1235-1237.
29. Hu L, Zhou M, Young A, Zhao W, Yan Z. In vivo effectiveness and safety of probiotics on prophylaxis and treatment of oral candidiasis: a systematic review and meta-analysis. BMC Oral Health 2019; 19: 140.
30. Srivastava N, Ellepola K, Venkiteswaran N, Chai LYA, Ohshima T, Seneviratne CJ. Lactobacillus Plantarum 108 Inhibits Streptococcus mutans and Candida albicans Mixed-Species Biofilm Formation. Antibiotics (Basel) 2020; 9: 478.
31. Ai R, Wei J, Ma D, Jiang L, Dan H, Zhou Y, et al. A meta-analysis of randomized trials assessing the effects of probiotic preparations on oral candidiasis in the elderly. Arch Oral Biol 2017; 83: 187-192.
32. Zeng Y, Fadaak A, Alomeir N, Wu Y, Wu TT, Qing S, et al. Effect of Probiotic Lactobacillus plantarum on Streptococcus mutans and Candida albicans Clinical Isolates from Children with Early Childhood Caries. Int J Mol Sci 2023; 24: 2991.
33. Rather IA, Wani MY, Kamli MR, Sabir JSM, Hakeem KR, Firoz A, et al. Limosilactobacillus fermentum KAU0021 Abrogates Mono- and Polymicrobial Biofilms Formed by Candida albicans and Staphylococcus aureus. Pharmaceutics 2023; 15: 1079.
34. Peeters E, Nelis HJ, Coenye T. Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates. J Microbiol Methods 2008; 72: 157-165.
35. Stepanović S, Vuković D, Hola V, Di Bonaventura G, Djukić S, Cirković I, et al. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS 2007; 115: 891-899.
36. Chandra J, Kuhn DM, Mukherjee PK, Hoyer LL, McCormick T, Ghannoum MA. Biofilm formation by the fungal pathogen Candida albicans: development, architecture, and drug resistance. J Bacteriol 2001; 183: 5385-5394.
37. Jakubovics NS, Goodman SD, Mashburn-Warren L, Stafford GP, Cieplik F. The dental plaque biofilm matrix. Periodontol 2000 2021; 86: 32-56.
| Files | ||
| Issue | Vol 18 No 5 (2026) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v18i5.22872 | |
| Keywords | ||
| Antifungal agents Biofilms Candida albicans Probiotics Lactobacillus casei Nystatin | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Faryabi O, Isaei E, Salari S, Ayatollahi Mousavi SA, Salarkia E, Sobhanipoor MH, Ghasemi Nejad Almani P, Izadi A. Evaluation of antifungal and antibiofilm effects of supernatant cell-free supernatant (CFS) of Lactobacillus casei alone and in combination with nystatin against Candida albicans strains. Iran J Microbiol. 2026;18(5):818-827.
