Anti-biofilm mechanisms of 3,5-di-tert-butylphenol against clinically relevant fungal pathogens

Abstract

The methanolic extract (PFME) of Pleurotus florida was assessed for anti-biofilm activity against Candida species. 3,5-Di-tert-butylphenol (3,5-DTB) was identified as the major antifungal constituent in PFME. In its pure form 3,5-DTB inhibits, disrupts, and reduces the viability of biofilm cells as seen from scanning electron and confocal microscopy studies. Microscopic studies and propidium iodide uptake assays confirmed that 3,5-DTB damages the cell membrane of Candida cells. In addition, 3,5-DTB induces accumulation of reactive oxygen species (ROS) which contribute to its pronounced anti-biofilm activity. The results of the present study show that 3,5-DTB exhibits combined anti-biofilm and conventional fungicidal activity against Candida species and elucidate the underlying mechanisms.

Synergistic Effect of Quinic Acid Derived From Syzygium cumini and Undecanoic Acid Against Candida spp. Biofilm and Virulence

“…Further, Light and CLSM micrographs visibly evidenced that hyphal elongation, microcolony formation, biofilm thickness were considerably decreased by QA-UDA combination. In the same way, usnic acid and 3,5-Di-tert-butylphenol diminished the hyphal growth and biofilm thickness of C. albicans and C. tropicalis ( Nithyanand et al, 2015 ; Rathna et al, 2016 ).…”

Antifungal Effects of Saponin Extract from Rhizomes of Dioscorea panthaica Prain et Burk against Candida albicans

“…The fluorescent dye PI has been widely used to detect the membrane permeability because this membrane impermeable probe could only enter cells with permeability-compromised membrane [ 24 , 34 , 45 ]. FCM and CLSM assay showed that HSE caused significant damage to the plasma membrane of C. albicans cells, while the membrane-perturbation effects of HSE are akin to many other antifungal agents, for example, antimicrobial peptides, diphenyl diselenide, 3,5-di-tert-butylphenol, hibicuslide C, and thymoquinone [ 24 , 45 – 48 ].…”

“…For quantitative analysis, following treatment and incubation with PI, cells were subjected to flow cytometry (FCM) (Beckman Coulter EPICS XL-MCL, USA) equipped with an argon laser (488 nm) for excitation to detect the fluorescent intensity of cells [ 24 ]. The percentage of PI stained cells was analyzed by Expo32 ADC analysis software (Beckman Coulter EPICS XL, USA).…”

“…As for ROS production in mature biofilms in 96-well plate, DCFH-DA was added to wells with biofilms to achieve a concentration of 10 μ M. After incubation for 20 minutes in dark at 37°C and removing excessive extracellular dye by washing, biofilms were incubated with indicated concentrations of HSE for 2 h in dark. The fluorescent intensity of biofilms was detected by VarioSkan multifunctional microplate reader at Ex 485 nm/Em 525 nm [ 24 ].…”

“…Before the viability assay, the morphologies of biofilms were photographed with microscope. The concentration of NAC used was 150 μ g/mL, according to the previous study performed by others [ 24 ]. These assays were performed in triplicate and repeated at least three times.…”