Fungal Biofilms, 2nd Edition

A special issue of Journal of Fungi (ISSN 2309-608X). This special issue belongs to the section "Fungal Cell Biology, Metabolism and Physiology".

Deadline for manuscript submissions: closed (15 July 2024) | Viewed by 9949

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Guest Editor
1. 1H-TOXRUN—One Health Toxicology Research Unit, CESPU-IUCS, Gandra, Portugal
2. LEPABE, University of Porto, Porto, Portugal
Interests: biofilms; fungal and bacterial infections; resistance to antimicrobials; microfluidics; pathogens detection; alternative therapies; surface functionalization of biomaterials
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Guest Editor
Department of Microbiology and Virology, Faculty of Natural Sciences, Comenius University in Bratislava, 842 15 Bratislava, Slovakia
Interests: biofilm; Candida; MRSA; virulence; resistance; farnesol; photodynamic inactivation
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Fungal infections are an important and increasing global threat, carrying not only high morbidity and mortality rates, but also high healthcare costs. Without an effective response, it is predicted that 10 million people will die per year as a result of multidrug-resistant pathogens. A high percentage of the mortalities caused by fungi are known to be biofilm-related.

This Special Issue, "Fungal Biofilms", is intended to cover the state of fungal biofilm research, from virulence and pathogenicity, to new compounds with antibiofilm and antifungal activity. We welcome reviews and original research articles covering the development/evaluation/validation of recent studies, especially those regarding multidrug resistance.

Dr. Célia F. Rodrigues
Dr. Lucia Černáková
Guest Editors

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Keywords

  • fungi
  • infection
  • biofilm
  • Candida
  • Aspergillus
  • Cryptococcus
  • antifungal
  • resistance
  • matrix

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Published Papers (7 papers)

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Research

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12 pages, 1384 KiB  
Article
Changes in Ergosterol Biosynthesis Alter the Response to Cycloheximide, 4-Nitroquinoline-N-Oxide, Weak Organic Acids, and Virulence in Candida glabrata
by Daniel Eliaš, Nora Tóth Hervay, Lucia Černáková and Yvetta Gbelská
J. Fungi 2024, 10(10), 669; https://doi.org/10.3390/jof10100669 - 26 Sep 2024
Viewed by 509
Abstract
The ERG6 gene encodes the sterol C24-methyltransferase converting zymosterol to fecosterol in the ergosterol biosynthetic pathway. Here, we extend the results of functional analysis of the CgERG6 gene, which was previously shown to modulate drug susceptibility in Candida glabrata mutant cells, by demonstrating [...] Read more.
The ERG6 gene encodes the sterol C24-methyltransferase converting zymosterol to fecosterol in the ergosterol biosynthetic pathway. Here, we extend the results of functional analysis of the CgERG6 gene, which was previously shown to modulate drug susceptibility in Candida glabrata mutant cells, by demonstrating that its deletion leads to increased susceptibility to cycloheximide, 4-nitroquinoline-N-oxide and weak organic acids, and such effects are associated with attenuated virulence. Together with abrogated efflux of drug substrates by CgCdr1p and CgPdr12p, the Cgerg6Δ mutation leads to reduced cell surface hydrophobicity and decreased virulence of the mutant cells of C. glabrata. The absence of CgErg6p impacts the lipid organization and function of the plasma membrane, resulting in non-specific permeability and abrogation of normal function of membrane-bound proteins accompanied by decreased virulence in Cgerg6Δ cells. Galleria mellonella larvae were used as a non-vertebrate animal host model to determine differences in the virulence potential of C. glabrata strains (parental strain and the Cgerg6Δ deletion mutant). We found that Cgerg6Δ mutant strain attenuated in virulence caused 25–30% survival of larvae compared with parental strain. Full article
(This article belongs to the Special Issue Fungal Biofilms, 2nd Edition)
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22 pages, 3224 KiB  
Article
Phenotypic and Genotypic Characterization of Resistance and Virulence Markers in Candida spp. Isolated from Community-Acquired Infections in Bucharest, and the Impact of AgNPs on the Highly Resistant Isolates
by Viorica Maria Corbu, Ana-Maria Georgescu, Ioana Cristina Marinas, Radu Pericleanu, Denisa Vasilica Mogos, Andreea Ștefania Dumbravă, Liliana Marinescu, Ionut Pecete, Tatiana Vassu-Dimov, Ilda Czobor Barbu, Ortansa Csutak, Denisa Ficai and Irina Gheorghe-Barbu
J. Fungi 2024, 10(8), 563; https://doi.org/10.3390/jof10080563 - 9 Aug 2024
Viewed by 813
Abstract
Background: This study aimed to determine, at the phenotypic and molecular levels, resistance and virulence markers in Candida spp. isolated from community-acquired infections in Bucharest outpatients during 2021, and to demonstrate the efficiency of alternative solutions against them based on silver nanoparticles (AgNPs). [...] Read more.
Background: This study aimed to determine, at the phenotypic and molecular levels, resistance and virulence markers in Candida spp. isolated from community-acquired infections in Bucharest outpatients during 2021, and to demonstrate the efficiency of alternative solutions against them based on silver nanoparticles (AgNPs). Methods: A total of 62 Candida spp. strains were isolated from dermatomycoses and identified using chromogenic culture media and MALDI-TOF MS, and then investigated for their antimicrobial resistance and virulence markers (VMs), as well as for metabolic enzymes using enzymatic tests for the expression of soluble virulence factors, their biofilm formation and adherence capacity on HeLa cells, and PCR assays for the detection of virulence markers and the antimicrobial activity of alternative solutions based on AgNPs. Results: Of the total of 62 strains, 45.16% were Candida parapsilosis; 29.03% Candida albicans; 9.67% Candida guilliermondii; 3.22% Candida lusitaniae, Candia pararugosa, and Candida tropicalis; and 1.66% Candida kefyr, Candida famata, Candida haemulonii, and Candida metapsilosis. Aesculin hydrolysis, caseinase, and amylase production were detected in the analyzed strains. The strains exhibited different indices of adherence to HeLa cells and were positive in decreasing frequency order for the LIP1, HWP1, and ALS1,3 genes (C. tropicalis/C. albicans). An inhibitory effect on microbial growth, adherence capacity, and on the production of virulence factors was obtained using AgNPs. Conclusions: The obtained results in C. albicans and Candida non-albicans circulating in Bucharest outpatients were characterized by moderate-to-high potential to produce VMs, necessitating epidemiological surveillance measures to minimize the chances of severe invasive infections. Full article
(This article belongs to the Special Issue Fungal Biofilms, 2nd Edition)
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20 pages, 72710 KiB  
Article
Influence of Zinc on Histoplasma capsulatum Planktonic and Biofilm Cells
by Ana Carolina Moreira da Silva Pires, Angélica Romão Carvalho, Carolina Orlando Vaso, Maria José Soares Mendes-Giannini, Junya de Lacorte Singulani and Ana Marisa Fusco-Almeida
J. Fungi 2024, 10(5), 361; https://doi.org/10.3390/jof10050361 - 20 May 2024
Viewed by 1136
Abstract
Histoplasma capsulatum causes a fungal respiratory disease. Some studies suggest that the fungus requires zinc to consolidate the infection. This study aimed to investigate the influence of zinc and the metal chelator TPEN on the growth of Histoplasma in planktonic and biofilm forms. [...] Read more.
Histoplasma capsulatum causes a fungal respiratory disease. Some studies suggest that the fungus requires zinc to consolidate the infection. This study aimed to investigate the influence of zinc and the metal chelator TPEN on the growth of Histoplasma in planktonic and biofilm forms. The results showed that zinc increased the metabolic activity, cell density, and cell viability of planktonic growth. Similarly, there was an increase in biofilm metabolic activity but no increase in biomass or extracellular matrix production. N′-N,N,N,N–tetrakis–2-pyridylmethylethane–1,2 diamine (TPEN) dramatically reduced the same parameters in the planktonic form and resulted in a decrease in metabolic activity, biomass, and extracellular matrix production for the biofilm form. Therefore, the unprecedented observations in this study highlight the importance of zinc ions for the growth, development, and proliferation of H. capsulatum cells and provide new insights into the role of metal ions for biofilm formation in the dimorphic fungus Histoplasma, which could be a potential therapeutic strategy. Full article
(This article belongs to the Special Issue Fungal Biofilms, 2nd Edition)
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19 pages, 2276 KiB  
Article
Central Carbon Metabolism in Candida albicans Biofilms Is Altered by Dimethyl Sulfoxide
by Maria Fernanda Cordeiro Arruda, Romeu Cassiano Pucci da Silva Ramos, Nicoly Subtil de Oliveira, Rosimeire Takaki Rosa, Patrícia Maria Stuelp-Campelo, Luiz Fernando Bianchini, Silas Granato Villas-Bôas and Edvaldo Antonio Ribeiro Rosa
J. Fungi 2024, 10(5), 337; https://doi.org/10.3390/jof10050337 - 8 May 2024
Viewed by 1165
Abstract
The effect of dimethyl sulfoxide (DMSO) on fungal metabolism has not been well studied. This study aimed to evaluate, by metabolomics, the impact of DMSO on the central carbon metabolism of Candida albicans. Biofilms of C. albicans SC5314 were grown on paper [...] Read more.
The effect of dimethyl sulfoxide (DMSO) on fungal metabolism has not been well studied. This study aimed to evaluate, by metabolomics, the impact of DMSO on the central carbon metabolism of Candida albicans. Biofilms of C. albicans SC5314 were grown on paper discs, using minimum mineral (MM) medium, in a dynamic continuous flow system. The two experimental conditions were control and 0.03% DMSO (v/v). After 72 h of incubation (37 °C), the biofilms were collected and the metabolites were extracted. The extracted metabolites were subjected to gas chromatography–mass spectrometry (GC/MS). The experiment was conducted using five replicates on three independent occasions. The GC/MS analysis identified 88 compounds. Among the 88 compounds, the levels of 27 compounds were markedly different between the two groups. The DMSO group exhibited enhanced levels of putrescine and glutathione and decreased levels of methionine and lysine. Additionally, the DMSO group exhibited alterations in 13 metabolic pathways involved in primary and secondary cellular metabolism. Among the 13 altered pathways, seven were downregulated and six were upregulated in the DMSO group. These results indicated a differential intracellular metabolic profile between the untreated and DMSO-treated biofilms. Hence, DMSO was demonstrated to affect the metabolic pathways of C. albicans. These results suggest that DMSO may influence the results of laboratory tests when it is used as a solvent. Hence, the use of DMSO as a solvent must be carefully considered in drug research, as the effect of the researched drugs may not be reliably translated into clinical practice. Full article
(This article belongs to the Special Issue Fungal Biofilms, 2nd Edition)
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14 pages, 2389 KiB  
Article
Died or Not Dyed: Assessment of Viability and Vitality Dyes on Planktonic Cells and Biofilms from Candida parapsilosis
by Betsy Verónica Arévalo-Jaimes and Eduard Torrents
J. Fungi 2024, 10(3), 209; https://doi.org/10.3390/jof10030209 - 11 Mar 2024
Viewed by 2009
Abstract
Viability and vitality assays play a crucial role in assessing the effectiveness of novel therapeutic approaches, with stain-based methods providing speed and objectivity. However, their application in yeast research lacks consensus. This study aimed to assess the performance of four common dyes on [...] Read more.
Viability and vitality assays play a crucial role in assessing the effectiveness of novel therapeutic approaches, with stain-based methods providing speed and objectivity. However, their application in yeast research lacks consensus. This study aimed to assess the performance of four common dyes on C. parapsilosis planktonic cells as well as sessile cells that form well-structured biofilms (treated and not treated with amphotericin B). Viability assessment employed Syto-9 (S9), thiazole orange (TO), and propidium iodide (PI). Metabolic activity was determined using fluorescein diacetate (FDA) and FUN-1. Calcofluor white (CW) served as the cell visualization control. Viability/vitality percentage of treated samples were calculated for each dye from confocal images and compared to crystal violet and PrestoBlue results. Heterogeneity in fluorescence intensity and permeability issues were observed with S9, TO, and FDA in planktonic cells and biofilms. This variability, influenced by cell morphology, resulted in dye-dependent viability/vitality percentages. Notably, PI and FUN-1 exhibited robust C. parapsilosis staining, with FUN-1 vitality results comparable to PrestoBlue. Our finding emphasizes the importance of evaluating dye permeability in yeast species beforehand, incorporating cell visualization controls. An improper dye selection may lead to misinterpreting treatment efficacy. Full article
(This article belongs to the Special Issue Fungal Biofilms, 2nd Edition)
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17 pages, 1447 KiB  
Article
High-Throughput Screening of the Repurposing Hub Library to Identify Drugs with Novel Inhibitory Activity against Candida albicans and Candida auris Biofilms
by Olabayo H. Ajetunmobi, Gina Wall, Bruna Vidal Bonifacio, Lucero A. Martinez Delgado, Ashok K. Chaturvedi, Laura K. Najvar, Floyd L. Wormley, Jr., Hoja P. Patterson, Nathan P. Wiederhold, Thomas F. Patterson and Jose L. Lopez-Ribot
J. Fungi 2023, 9(9), 879; https://doi.org/10.3390/jof9090879 - 27 Aug 2023
Cited by 3 | Viewed by 1870
Abstract
Candidiasis is one of the most frequent nosocomial infections affecting an increasing number of at-risk patients. Candida albicans remains the most frequent causative agent of candidiasis, but, in the last decade, C. auris has emerged as a formidable multi-drug-resistant pathogen. Both species are [...] Read more.
Candidiasis is one of the most frequent nosocomial infections affecting an increasing number of at-risk patients. Candida albicans remains the most frequent causative agent of candidiasis, but, in the last decade, C. auris has emerged as a formidable multi-drug-resistant pathogen. Both species are fully capable of forming biofilms, which contribute to resistance, increasing the urgency for new effective antifungal therapies. Repurposing existing drugs could significantly accelerate the development of novel therapies against candidiasis. Here, we have screened the Repurposing Hub library from the Broad Institute, containing over 6000 compounds, in search for inhibitors of C. albicans and C. auris biofilm formation. The primary screen identified 57 initial hits against C. albicans and 33 against C. auris. Confirmatory concentration-dependent assays were used to validate the activity of the initial hits and, at the same time, establish their anti-biofilm potency. Based on these results, ebselen, temsirolimus, and compound BAY 11-7082 emerged as the leading repositionable compounds. Subsequent experiments established their spectrum of antifungal activity against yeasts and filamentous fungi. In addition, their in vivo activity was examined in the murine models of hematogenously disseminated C. albicans and C. auris infections. Although promising, further in vitro and in vivo studies are needed to confirm their potential use for the therapy of candidiasis and possibly other fungal infections. Full article
(This article belongs to the Special Issue Fungal Biofilms, 2nd Edition)
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Review

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14 pages, 3605 KiB  
Review
Interactions between Bacteria and Aspergillus fumigatus in Airways: From the Mycobiome to Molecular Interactions
by Anne Debourgogne, Lorra Monpierre, Khadeeja Adam Sy, Isabel Valsecchi, Jean-Winoc Decousser and Françoise Botterel
J. Fungi 2023, 9(9), 900; https://doi.org/10.3390/jof9090900 - 1 Sep 2023
Cited by 2 | Viewed by 1725
Abstract
Interactions between different kingdoms of microorganisms in humans are common but not well described. A recent analysis of the mycobiome has described the presence of different fungi and their positive and/or negative interactions with bacteria and other fungi. In chronic respiratory diseases, these [...] Read more.
Interactions between different kingdoms of microorganisms in humans are common but not well described. A recent analysis of the mycobiome has described the presence of different fungi and their positive and/or negative interactions with bacteria and other fungi. In chronic respiratory diseases, these different microorganisms form mixed biofilms to live inside. The interactions between Gram-negative bacteria and filamentous fungi in these biofilms have attracted more attention recently. In this review, we analyse the microbiota of the respiratory tract of healthy individuals and patients with chronic respiratory disease. Additionally, we describe the regulatory mechanisms that rule the mixed biofilms of Aspergillus fumigatus and Gram-negative bacteria and the effects of this biofilm on clinical presentations. Full article
(This article belongs to the Special Issue Fungal Biofilms, 2nd Edition)
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