Antibiofilm Activity and Mechanism of Action of the Disinfectant Chloramine T on Candida spp., and Its Toxicity against Human Cells
Abstract
:1. Introduction
2. Results
2.1. Determination of the MIC and MFC Values
2.2. Effects on C. Albicans Growth Kinetics
2.3. Mechanism of Action of CAT
2.4. Effect of CAT on C. albicans Micromorphology
2.5. Effects of CAT on the Adherence, Formation and Reduction of C. Albicans Mature Biofilms
2.6. Cytotoxicity of CAT on Human Erythrocytes
3. Discussion
4. Materials and Methods
4.1. Microorganisms
4.2. Determination of the Minimum Inhibitory and Fungicidal Concentration (MIC)
4.3. Effects of CAT on Candida spp. Growth Kinetics
4.4. Mechanism(s) of Action of CAT
4.4.1. Action on Cell Wall Biosynthesis
4.4.2. Action on Cell Membrane Permeability
4.5. Effects of CAT on Fungal Micromorphology
4.6. Effects of CAT on C. Albicans Biofilms
4.6.1. Effects on Early Biofilm Adherence
4.6.2. Effects on Mature Biofilm Formation
4.6.3. Reduction of Preformed Mature Biofilm
4.7. Cytotoxic Effects of CAT on Human Erythrocytes
4.8. Statistical Analysis
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Sample Availability: Samples of the of chloramine T trihydrate (INLAB-CAS 7080-50-4, São Paulo, Brazil) are available from the authors. |
Strain | CAT | NaOCl | Nystatin | ||||||
---|---|---|---|---|---|---|---|---|---|
MIC | MFC | MFC/MIC Ratio | MIC | MFC | MFC/MIC Ratio | MIC | MFC | MFC/MIC Ratio | |
Candida albicans ATCC 60193 | 2.77 | 5.54 | 2 (Fungicidal) | 8.39 | 8.39 | 1 (Fungicidal) | 0.0004 | 0.0008 | 2 (Fungicidal) |
Candida albicans CBS 562 | 0.69 | 2.77 | 4 (Fungistatic) | 4.19 | 8.39 | 2 (Fungicidal) | 0.0004 | 0.0008 | 2 (Fungicidal) |
Candida tropicalis ATCC 750 | 2.77 | 2.77 | 1 (Fungicidal) | 8.39 | 8.39 | 1 (Fungicidal) | 0.0004 | 0.0004 | 1 (Fungicidal) |
Candida tropicalis CBS 94 | 5.54 | 5.54 | 1 (Fungicidal) | 16.79 | 16.79 | 1 (Fungicidal) | 0.0004 | 0.0004 | 1 (Fungicidal) |
Candida krusei ATCC 3413 | 1.38 | 1.38 | 1 (Fungicidal) | 4.19 | 4.19 | 1 (Fungicidal) | 0.001 | 0.001 | 1 (Fungicidal) |
Candida krusei CBS 73 | 1.38 | 1.38 | 1 (Fungicidal) | 4.19 | 4.19 | 1 (Fungicidal) | 0.0008 | 0.0008 | 1 (Fungicidal) |
Candida glabrata IZ 07 | 5.54 | 11.09 | 2 (Fungicidal) | 16.79 | 16.79 | 1 (Fungicidal) | 0.0004 | 0.0008 | 2 (Fungicidal) |
Strain | MIC (mmol/L) | |||||
---|---|---|---|---|---|---|
CAT | NaOCl | Caspofungin | ||||
Without Sorbitol | With Sorbitol | Without Sorbitol | With Sorbitol | Without Sorbitol | With Sorbitol | |
Candida albicans ATCC 60193 | 2.77 | 22.18 | 8.39 | 33.58 | <0.0003 | >0.0045 |
Candida albicans CBS 562 | 0.69 | 11.09 | 4.19 | 33.58 | <0.0003 | 0.0045 |
CAT | NaOCl | Nystatin | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Absence of Ergosterol | Presence of Ergosterol | Absence of Ergosterol | Presence of Ergosterol | Absence of Ergosterol | Presence of Ergosterol | |||||||
100 µg/mL | 200 µg/mL | 400 µg/mL | 100 µg/mL | 200 µg/mL | 400 µg/mL | 100 µg/mL | 200 µg/mL | 400 µg/mL | ||||
Candida albicans ATCC 60193 | 2.77 | 5.54 | 5.54 | 5.54 | 8.39 | 16.79 | 16.79 | 16.79 | 0.0004 | 0.001 | 0.003 | 0.003 |
Candida albicans CBS 562 | 0.69 | 5.54 | 5.54 | 5.54 | 4.19 | 16.79 | 33.58 | 33.58 | 0.0004 | 0.001 | 0.001 | 0.001 |
Concentration | Inhibition of Initial Adherence | Inhibition Of Mature Biofilm Formation | ||||
---|---|---|---|---|---|---|
Group 1 (Exposure time: 2 h) | Group 2 (Exposure time: 1 min) | Group 3 (Exposure time: 8 h) | ||||
CAT | NaOCl | CAT | NaOCl | CAT | NaOCl | |
MIC | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa | %I ≤ 25% Aa | 25% < %I ≤ 50% Aa |
2 × MIC | %I ≤ 25% Aa | 50% < %I ≤ 75% Ba | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa |
4 × MIC | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa |
Concentration | Group 4 (Exposure time: 3 × 1 min) | Group 5 (Exposure Time: 6 × 1 min) | Group 6 (Exposure time: 8 h) | Group 7 (Exposure time: 2 × 8 h) | ||||
---|---|---|---|---|---|---|---|---|
CAT | NaOCl | CAT | NaOCl | CAT | NaOCl | CAT | NaOCl | |
MIC | 50% < %I ≤ 75% Aa | %I ≤ 25% Aa | %I ≤ 25% Aa | %I ≤ 25% Aa | 50% < %I ≤ 75% Aa | 50% < %I ≤ 75% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa |
2 × MIC | 50% < %I ≤ 75% Aa | %I ≤ 25% Ba | %I ≤ 25% Aa | %I ≤ 25% Aa | 50% < %I ≤ 75% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa |
4 × MIC | 50%< %I ≤ 75% Aa | %I ≤ 50% Aa | 25% <%I ≤ 50% Aa | %I ≤ 25% Ba | 50% < %I ≤ 75% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa | 25% < %I ≤ 50% Aa |
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Ferreira, G.L.S.; Rosalen, P.L.; Peixoto, L.R.; Pérez, A.L.A.d.L.; Carlo, F.G.d.C.; Castellano, L.R.C.; Lima, J.M.d.; Freires, I.A.; Lima, E.D.O.; Castro, R.D.d. Antibiofilm Activity and Mechanism of Action of the Disinfectant Chloramine T on Candida spp., and Its Toxicity against Human Cells. Molecules 2017, 22, 1527. https://doi.org/10.3390/molecules22091527
Ferreira GLS, Rosalen PL, Peixoto LR, Pérez ALAdL, Carlo FGdC, Castellano LRC, Lima JMd, Freires IA, Lima EDO, Castro RDd. Antibiofilm Activity and Mechanism of Action of the Disinfectant Chloramine T on Candida spp., and Its Toxicity against Human Cells. Molecules. 2017; 22(9):1527. https://doi.org/10.3390/molecules22091527
Chicago/Turabian StyleFerreira, Gabriela Lacet Silva, Pedro Luiz Rosalen, Larissa Rangel Peixoto, Ana Luiza Alves de Lima Pérez, Fabíola Galbiatti de Carvalho Carlo, Lúcio Roberto Cançado Castellano, Jefferson Muniz de Lima, Irlan Almeida Freires, Edeltrudes De Oliveira Lima, and Ricardo Dias de Castro. 2017. "Antibiofilm Activity and Mechanism of Action of the Disinfectant Chloramine T on Candida spp., and Its Toxicity against Human Cells" Molecules 22, no. 9: 1527. https://doi.org/10.3390/molecules22091527
APA StyleFerreira, G. L. S., Rosalen, P. L., Peixoto, L. R., Pérez, A. L. A. d. L., Carlo, F. G. d. C., Castellano, L. R. C., Lima, J. M. d., Freires, I. A., Lima, E. D. O., & Castro, R. D. d. (2017). Antibiofilm Activity and Mechanism of Action of the Disinfectant Chloramine T on Candida spp., and Its Toxicity against Human Cells. Molecules, 22(9), 1527. https://doi.org/10.3390/molecules22091527