Temporal Profile of Biofilm Formation, Gene Expression and Virulence Analysis in Candida albicans Strains

ABSTRACT Candida albicans biofilms are structured microbial communities with high levels of drug resistance. Farnesol, a quorum-sensing molecule that inhibits hyphal formation in C. albicans, has been found to prevent biofilm formation by C. albicans. There is limited information, however, about the molecular mechanism of farnesol against biofilm formation. We used cDNA microarray analysis to identify the changes in the gene expression profile of a C. albicans biofilm inhibited by farnesol. Confocal scanning laser microscopy was used to visualize and confirm normal and farnesol-inhibited biofilms. A total of 274 genes were identified as responsive, with 104 genes up-regulated and 170 genes down-regulated. Independent reverse transcription-PCR analysis was used to confirm the important changes detected by microarray analysis. In addition to hyphal formation-associated genes (e.g., TUP1, CRK1, and PDE2), a number of other genes with roles related to drug resistance (e.g., FCR1 and PDR16), cell wall maintenance (e.g., CHT2 and CHT3), and iron transport (e.g., FTR2) were responsive, as were several genes encoding heat shock proteins (e.g., HSP70, HSP90, HSP104, CaMSI3, and SSA2). Further study of these differentially regulated genes is warranted to evaluate how they may be involved in C. albicans biofilm formation. Consistent with the down-regulation of the cell surface hydrophobicity-associated gene (CSH1), the water-hydrocarbon two-phase assay showed a decrease in cell surface hydrophobicity in the farnesol-treated group compared to that in the control group. Our data provide new insight into the molecular mechanism of farnesol against C. albicans biofilm formation.

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Propolis ethanolic extract influenced biofilm formation and gene expression level of EFG1 in Candida albicans

10.1063/5.0052928 ◽

2021 ◽

Author(s):

Sumarno Reto Prawiro ◽

A. A. Santi Dewi ◽

Anggie Diniayuningrum ◽

Dinda Oktia Maghfiroh ◽

Nurdiana Nurdiana ◽

...

Keyword(s):

Gene Expression ◽

Candida Albicans ◽

Biofilm Formation ◽

Ethanolic Extract ◽

Gene Expression Level ◽

Expression Level

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ALS3 Expression as an Indicator for Candida albicans Biofilm Formation and Drug Resistance

Frontiers in Microbiology ◽

10.3389/fmicb.2021.655242 ◽

2021 ◽

Vol 12 ◽

Author(s):

Keke Deng ◽

Wei Jiang ◽

Yanyu Jiang ◽

Qi Deng ◽

Jinzhong Cao ◽

...

Keyword(s):

Gene Expression ◽

Candida Albicans ◽

Biofilm Formation ◽

Antifungal Agents ◽

Early Prevention ◽

Therapy Failure ◽

Prevention And Treatment ◽

Selection Of ◽

Prevention And Therapy

Resistance caused by the formation of the Candida albicans (C. albicans) biofilm is one of the main reasons for antifungal therapy failure. Thus, it is important to find indicators that predict C. albicans biofilm formation to provide evidence for the early prevention and treatment of the C. albicans biofilms. In this study, C. albicans samples were selected from C. albicans septicemia that were sensitive to common antifungal agents. It was found that the agglutinin-like sequence 3 (ALS3) gene was differentially expressed in free, antifungal, drug-sensitive C. albicans. The average ALS3 gene expression was higher in the C. albicans strains with biofilm formation than that in the C. albicans strains without biofilm formation. Then, it was further confirmed that the rate of biofilm formation was higher in the high ALS3 gene expression group than that in the low ALS3 gene expression group. It was found that C. albicans with biofilm formation was more resistant to fluconazole, voriconazole, and itraconazole. However, it maintained its sensitivity to caspofungin and micafungin in vitro and in mice. Further experiments regarding the prevention of C. albicans biofilm formation were performed in mice, in which only caspofungin and micafungin prevented C. albicans biofilm formation. These results suggest that the expression level of ALS3 in C. albicans may be used as an indicator to determine whether C. albicans will form biofilms. The results also show that the biofilm formation of C. albicans remained sensitive to caspofungin and micafungin, which may help to guide the selection of clinical antifungal agents for prevention and therapy.

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Effects of Bacillus subtilis on Candida albicans: biofilm formation, filamentation and gene expression

Brazilian Dental Science ◽

10.14295/bds.2019.v22i2.1692 ◽

2019 ◽

Vol 22 (2) ◽

pp. 252-259

Author(s):

Michelle Peneluppi Silva ◽

Patrícia Pimentel De Barros ◽

Adeline Lacerda Jorjão ◽

Rodnei Dennis Rossoni ◽

Juliana Campos Junqueira ◽

...

Keyword(s):

Gene Expression ◽

Bacillus Subtilis ◽

Candida Albicans ◽

Real Time ◽

Real Time Pcr ◽

Bovine Serum ◽

Biofilm Formation ◽

Fetal Bovine Serum ◽

Viable Cells ◽

Fetal Bovine

Objective: The aim of this study was evaluate the effect of Bacillus subtilis on Candida albicans biofilm formation and filamentation by evaluating the gene expression of ALS3, HWP1, BCR1, EFG1 and TEC1. Material and Methods: Mixed (C. albicans / B.subtilis) and monotypic biofilms were cultured in plates at 37°C for 48 h under shaking for counting viable cells (CFU / mL) and analysis of gene expression by real-time PCR. The C. albicans filamentation assay was performed in medium containing 10% fetal bovine serum at 37°C for 6 hours. Data was analysed by t-Student and Mann– Whitney tests. Results: B. subtilis reduced the biofilm formation of C. albicans in 1 log when cultured in the same environment (p<0.0001). In addition, it significantly reduced the yeast -hypha transition affecting the morphology of C. albicans. Among all of the analyzed genes, the ALS3 and HWP1 genes were the most affected, achieving 111.1- and 333.3- fold decreases in the C. albicans biofilms associated with B. subtilis, respectively. Conclusion: B. subtilis reduced the biofilm formation and filamentation of C. albicans by negatively regulating the ALS3, HWP1, BCR1, EFG1 and TEC1 genes that are essential for the production of biofilm and hyphae.KeywordsBacillus subtilis; Candida albicans; Biofilm; Filamentation; Gene expression.

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Thidiazuron, a phenyl-urea cytokinin, inhibits ergosterol synthesis and attenuates biofilm formation of Candida albicans

10.21203/rs.3.rs-780442/v1 ◽

2021 ◽

Author(s):

Harikrishnan Pandurangan ◽

Balamani Arayambath ◽

Vijay Karthik Jayaraman ◽

Kanimozhi Ekambaram ◽

Emad A Ahmed ◽

...

Keyword(s):

Gene Expression ◽

Candida Albicans ◽

Crystal Violet ◽

Biofilm Formation ◽

Reduction Assay ◽

Ergosterol Synthesis ◽

Inhibitory Potential ◽

Anti Fungal ◽

Time Kill

Abstract Candida albicans is a commensal human fungal pathogen that colonizes and develops dental biofilm which cause Oral candidiosis. This study investigates the effects of a new molecule Thidiazuron against the growth and biofilm formation properties of C. albicans. This study applied computational and in vitro approaches such as broth microdilution, SEM, time-kill dynamics, crystal violet assay, XTT reduction assay, ergosterol quantification and quantitative RT PCR analysis of gene expression to validate the growth and biofilm inhibitory potential of thidiazuron against C. albicans. Preliminary molecular docking study revealed potential interaction between thidiazuron and amino acids residues of CYP51. Further in vitro anti-fungal susceptibility test, SEM and time kill analysis revealed anti-fungal potency of thidiazuron in dose and time dependent passion. Crystal violet staining, XTT reduction assay and Acridine Orange staining visually confirmed biofilm inhibitory potential of thidiazuron. Gene expression study shows that thidiazuron treatment down regulated the expression of genes involved in ergosterol synthesis, cell adhesion and hyphae development in C. albicans. This study identified thidiazuron as CYP51 inhibitor and a new antibiofilm agent against C. albicans.

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Role of cell surface hydrophobicity in Candida albicans biofilm

Open Life Sciences ◽

10.2478/s11535-013-0136-y ◽

2013 ◽

Vol 8 (3) ◽

pp. 259-262 ◽

Cited By ~ 9

Author(s):

Helena Bujdáková ◽

Miroslava Didiášová ◽

Hana Drahovská ◽

Lucia Černáková

Keyword(s):

Gene Expression ◽

Candida Albicans ◽

Cell Surface ◽

Biofilm Formation ◽

Cell Surface Hydrophobicity ◽

Surface Hydrophobicity ◽

Quantitative Real Time Pcr ◽

Planktonic Cells ◽

Planktonic Culture

AbstractOverall cell surface hydrophobicity (CSH) is predicted to play an important role during biofilm formation in Candida albicans but is the result of many expressed proteins. This study compares the CSH status and CSH1 gene expression in C. albicans planktonic cells, sessile biofilm, and dispersal cells. Greater percentages of hydrophobic cells were found in non-adhered (1.5 h) and dispersal forms (24 or 48 h) (41.34±4.17% and 39.52±7.45%, respectively), compared with overnight planktonic cultures (21.69±3.60%). Results from quantitative real-time PCR confirmed greater up-regulation of the CSH1 gene in sessile biofilm compared with both planktonic culture and dispersal cells. Up-regulation was also greater in dispersal cells compared with planktonic culture. The markedly increased CSH found both in C. albicans biofilm, and in cells released during biofilm formation could provide an advantage to dispersing cells building new biofilm.

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Relative Abundances ofCandida albicansandCandida glabratainIn VitroCoculture Biofilms Impact Biofilm Structure and Formation

Applied and Environmental Microbiology ◽

10.1128/aem.02769-17 ◽

2018 ◽

Vol 84 (8) ◽

pp. e02769-17 ◽

Cited By ~ 9

Author(s):

Michelle L. Olson ◽

Arul Jayaraman ◽

Katy C. Kao

Keyword(s):

Gene Expression ◽

Candida Albicans ◽

Biofilm Formation ◽

Candida Glabrata ◽

Interspecific Interactions ◽

Host Immune System ◽

Dependent Manner ◽

Important Species ◽

Content Type ◽

Content Ratio

ABSTRACTCandidais a member of the normal human microbiota and often resides on mucosal surfaces such as the oral cavity or the gastrointestinal tract. In addition to their commensality,Candidaspecies can opportunistically become pathogenic if the host microbiota is disrupted or if the host immune system becomes compromised. An important factor forCandidapathogenesis is its ability to form biofilm communities. The two most medically important species—Candida albicansandCandida glabrata—are often coisolated from infection sites, suggesting the importance ofCandidacoculture biofilms. In this work, we report that biofilm formation of the coculture population depends on the relative ratio of starting cell concentrations ofC. albicansandC. glabrata. When using a starting ratio ofC. albicanstoC. glabrataof 1:3, ∼6.5- and ∼2.5-fold increases in biofilm biomass were observed relative to those of aC. albicansmonoculture and aC. albicans/C. glabrataratio of 1:1, respectively. Confocal microscopy analysis revealed the heterogeneity and complex structures composed of longC. albicanshyphae andC. glabratacell clusters in the coculture biofilms, and reverse transcription-quantitative PCR (qRT-PCR) studies showed increases in the relative expression of theHWP1andALS3adhesion genes in theC. albicans/C. glabrata1:3 biofilm compared to that in theC. albicansmonoculture biofilm. Additionally, only the 1:3C. albicans/C. glabratabiofilm demonstrated an increased resistance to the antifungal drug caspofungin. Overall, the results suggest that interspecific interactions between these two fungal pathogens increase biofilm formation and virulence-related gene expression in a coculture composition-dependent manner.IMPORTANCECandida albicansandCandida glabrataare often coisolated during infection, and the occurrence of coisolation increases with increasing inflammation, suggesting possible synergistic interactions between the twoCandidaspecies in pathogenesis. During the course of an infection, the prevalence of eachCandidaspecies may change over time due to differences in metabolism and in the resistance of each species to antifungal therapies. Therefore, it is necessary to understand the dynamics betweenC. albicansandC. glabratain coculture to develop better therapeutic strategies againstCandidainfections. Existingin vitrowork has focused on understanding how an equal-part culture ofC. albicansandC. glabrataimpacts biofilm formation and pathogenesis. What is not understood, and what is investigated in this work, is how the composition ofCandidaspecies in coculture impacts overall biofilm formation, virulence gene expression, and the therapeutic treatment of biofilms.

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Monitoring ALS1 and ALS3 Gene Expression During In Vitro Candida albicans Biofilm Formation Under Continuous Flow Conditions

Mycopathologia ◽

10.1007/s11046-008-9148-6 ◽

2008 ◽

Vol 167 (1) ◽

pp. 9-17 ◽

Cited By ~ 28

Author(s):

Heleen Nailis ◽

Roosmarijn Vandenbroucke ◽

Kelly Tilleman ◽

Dieter Deforce ◽

Hans Nelis ◽

...

Keyword(s):

Gene Expression ◽

Candida Albicans ◽

Continuous Flow ◽

Biofilm Formation ◽

Flow Conditions

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Anticandidal Potential of Two Cyanobacteria-Synthesized Silver Nanoparticles: Effects on Growth, Cell Morphology, and Key Virulence Attributes of Candida albicans

Pharmaceutics ◽

10.3390/pharmaceutics13101688 ◽

2021 ◽

Vol 13 (10) ◽

pp. 1688

Author(s):

Reham Samir Hamida ◽

Mohamed Abdelaal Ali ◽

Doaa A. Goda ◽

Alya Redhwan

Keyword(s):

Gene Expression ◽

Silver Nanoparticles ◽

Candida Albicans ◽

Biofilm Formation ◽

Cell Envelope ◽

Membrane Integrity ◽

Ultrastructural Changes ◽

Protein Patterns ◽

Antimicrobial Drugs ◽

Fungal Cell

Candida albicans is an opportunistic human fungal pathogen responsible for 90–100% of mucosal and nosocomial infections worldwide. The emergence of drug-resistant strains has resulted in adverse consequences for human health, including numerous deaths. Consequently, there is an urgent need to identify and develop new antimicrobial drugs to counter these effects. Antimicrobial nanoagents have shown potent inhibitory activity against a number of pathogens through targeting their defense systems, such as biofilm formation. Here, we investigated the anticandidal activity of silver nanoparticles biosynthesized by the cyanobacterial strains Desertifilum sp. IPPAS B-1220 and Nostoc Bahar_M (D-SNPs and N-SNPs, respectively), along with that of silver nitrate (AgNO3), and examined the mechanisms underlying their lethal effects. For this, we performed agar well diffusion and enzyme activity assays (lactate dehydrogenase, adenosine triphosphatase, glutathione peroxidase, and catalase) and undertook morphological examinations using transmission electron microscopy. The effects of the three treatments on Hwp1 and CDR1 gene expression and protein patterns were assessed using qRT-PCR and SDS–PAGE assays, respectively. All of the three treatments inhibited C. albicans growth; disrupted membrane integrity, metabolic function, and antioxidant activity; induced ultrastructural changes in the cell envelope; and disrupted cytoplasmic and nuclear contents. Of the three agents, D-SNPs showed the greatest biocidal activity against C. albicans. Additionally, the D-SNP treatment significantly reduced the gene expression of Hwp1 and CDR1, suggestive of negative effects on biofilm formation ability and resistance potential of C. albicans, and promoted protein degradation. The mechanism involved in the biocidal effects of both D-SNPs and N-SNPs against C. albicans could be attributed to their ability to interfere with fungal cell structures and/or stimulate oxidative stress, enabling them to be used as a robust antimycotic agent.

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