Optimization of resazurin-based viability staining for quantification of microbial biofilms

Driessche, Freija Van den; Rigole, Petra; Brackman, Gilles; Coenye, Tom

doi:10.1016/j.mimet.2013.12.011

Cited by 104 publications

(81 citation statements)

References 12 publications

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“…A C. albicans SC5314 overnight culture, grown in YPD, was diluted to an optical density (OD) of 0.1 (approximately 10 6 cells/ml) in RPMI medium, and 100 l of this suspension was added to the wells of a round-bottom microplate (TPP Techno Plastic Products AG, Switzerland) (30,31). After 1 h of adhesion at 37°C, the medium was aspirated, and the biofilms were washed with 100 l phosphate-buffered saline (PBS) to remove nonadherent cells, followed by addition of 100 l RPMI 1640 medium.…”

Section: Methodsmentioning

confidence: 99%

Artemisinins, New Miconazole Potentiators Resulting in Increased Activity against Candida albicans Biofilms

Cremer

Lanckacker

Cools

et al. 2015

Antimicrob Agents Chemother

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cMucosal biofilm-related fungal infections are very common, and the incidence of recurrent oral and vulvovaginal candidiasis is significant. As resistance to azoles (the preferred treatment) is occurring, we aimed at identifying compounds that increase the activity of miconazole against Candida albicans biofilms. We screened 1,600 compounds of a drug-repositioning library in combination with a subinhibitory concentration of miconazole. Synergy between the best identified potentiators and miconazole was characterized by checkerboard analyses and fractional inhibitory concentration indices. Hexachlorophene, pyrvinium pamoate, and artesunate act synergistically with miconazole in affecting C. albicans biofilms. Synergy was most pronounced for artesunate and structural homologues thereof. No synergistic effect could be observed between artesunate and fluconazole, caspofungin, or amphotericin B. Our data reveal enhancement of the antibiofilm activity of miconazole by artesunate, pointing to potential combination therapy consisting of miconazole and artesunate to treat C. albicans biofilm-related infections.

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Section: Methodsmentioning

confidence: 99%

Artemisinins, New Miconazole Potentiators Resulting in Increased Activity against Candida albicans Biofilms

Cremer

Lanckacker

Cools

et al. 2015

Antimicrob Agents Chemother

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“…The control treatment was 1% ethanol in RPMI 1640 medium. The biofilms were washed with PBS, and biofilm formation was determined using CellTiter-Blue (Promega, Madison, WI) staining (38,39). After 1-h incubation at 37°C, fluorescence was measured with a multimode microplate reader, Synergy Mx (BioTek), at an excitation wavelength of 535 nm and emission wavelength of 590 nm.…”

Section: Methodsmentioning

confidence: 99%

Synergistic Activity of the Tyrocidines, Antimicrobial Cyclodecapeptides from Bacillus aneurinolyticus, with Amphotericin B and Caspofungin against Candida albicans Biofilms

Troskie

Rautenbach

Delattin

et al. 2014

Antimicrob Agents Chemother

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dTyrocidines are cationic cyclodecapeptides from Bacillus aneurinolyticus that are characterized by potent antibacterial and antimalarial activities. In this study, we show that various tyrocidines have significant activity against planktonic Candida albicans in the low-micromolar range. These tyrocidines also prevented C. albicans biofilm formation in vitro. Studies with the membrane-impermeable dye propidium iodide showed that the tyrocidines disrupt the membrane integrity of mature C. albicans biofilm cells. This membrane activity correlated with the permeabilization and rapid lysis of model fungal membranes containing phosphatidylcholine and ergosterol (70:30 ratio) induced by the tyrocidines. The tyrocidines exhibited pronounced synergistic biofilm-eradicating activity in combination with two key antifungal drugs, amphotericin B and caspofungin. Using a Caenorhabditis elegans infection model, we found that tyrocidine A potentiated the activity of caspofungin. Therefore, tyrocidines are promising candidates for further research as antifungal drugs and as agents for combinatorial treatment.

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“…at 37 uC for 24 and 48 h. After incubation, the biofilm formation was measured using a crystal violet staining method according to our previous study to detect polymeric substances (Majtan et al, 2014). The viability of biofilm-forming cells was measured using resazurin according to van den Driessche et al (2014).…”

Section: Pcrmentioning

confidence: 99%

Antibiofilm efficacy of honey and bee-derived defensin-1 on multispecies wound biofilm

Sojka

Valachová

Bucekova

et al. 2016

Journal of Medical Microbiology

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Many clinically relevant biofilms are polymicrobial. Examining the effect of antimicrobials in a multispecies biofilm consortium is of great clinical importance. The goal of this study was to investigate the effect of different honey types against bacterial wound pathogens grown in multispecies biofilm and to test the antibiofilm activity of honey defensin-1 (Def-1) in its recombinant form. A modified Lubbock chronic wound biofilm formed by four bacterial species (Staphylococcus aureus, Streptococcus agalactiae, Pseudomonas aeruginosa and Enterococcus faecalis) was used for evaluation of honey and recombinant bee-derived Def-1 antibiofilm efficacy. Recombinant Def-1 was prepared by heterologous expression in Escherichia coli. We showed that different types of honey (manuka and honeydew) were able to significantly reduce the cell viability of wound pathogens (Staphylococcus aureus, Streptococcus agalactiae and Pseudomonas aeruginosa) in mature polymicrobial biofilm. None of the tested honeys showed the ability to eradicate Enterococcus faecalis in biofilm. In addition, recombinant Def-1 successfully reduced the viability of Staphylococcus aureus and Pseudomonas aeruginosa cells within established polymicrobial biofilm after 24 and 48 h of treatment. Interestingly, recombinant Def-1 did not affect the viability of Streptococcus agalactiae cells within the biofilm, whereas both natural honeys significantly reduced the viable bacteria. Although Enterococcus faecalis was highly resistant to Def-1, Def-1 significantly affected the biofilm formation of Enterococcus faecalis and Streptococcus agalactiae after 24 h of treatment, most likely by inhibiting its extracellular polymeric substances production. In conclusion, our study revealed that honey and Def-1 are effective against established multispecies biofilm; however, Enterococcus faecalis grown in multispecies biofilm was resistant to both antimicrobials.

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Optimization of resazurin-based viability staining for quantification of microbial biofilms

Cited by 104 publications

References 12 publications

Artemisinins, New Miconazole Potentiators Resulting in Increased Activity against Candida albicans Biofilms

Artemisinins, New Miconazole Potentiators Resulting in Increased Activity against Candida albicans Biofilms

Synergistic Activity of the Tyrocidines, Antimicrobial Cyclodecapeptides from Bacillus aneurinolyticus, with Amphotericin B and Caspofungin against Candida albicans Biofilms

Antibiofilm efficacy of honey and bee-derived defensin-1 on multispecies wound biofilm

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