The discrepancy between Clove and Non-Clove Cigarette Smoke-Promoted Candida albicans Biofilm Formation with precoating RNA-aptamer

This study explores the influence of precoating aptamer (Ca-apt1) on C. albicans viability while the fungus was growing in the presence of exposing condensed cigarette smoke (CSC), prepared from clove (CCSC) and non-clove (NCSC) cigarettes, for 48 h. Using qPCR, we found that mRNA expression of adhesion-associated genes (ALS3 and HWP1) was impaired by precoating C. albicans yeast cells with the aptamer. Conversely, the gene transcription was upregulated when aptamer-uncoated yeast was pre-treated with either CSC. In addition, by analysing the result of MTT ([3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] assay, we found that the presence of added CCSC or NCSC in growth medium for 48 h was significantly enhanced C. albicans biofilm development. However, the presence of precoated aptamer was significantly impaired biofilm development accelerated by the NCSC. The inhibitory effect of the Ca-apt1 was not dependent on the precoated aptamer (1 and 10%). Interestingly, we noted that the enhancer effect of treated CCSC was no longer effective when the yeast had been precoated with 10% aptamer tested. Additionally, light microscopy analysis revealed that precoating aptamer alleviates morphological changes of C. albicans (from yeast to hypha formation) that are enhanced by adding CCSC or NCSC in the growth medium. In conclusion, these results suggest that administration on Ca-ap1 exhibits a significant protective effect on CSC-induced biofilm formation by C. albicans.

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The Discrepancy between Clove and Non-Clove Cigarette Smoke-Promoted Candida albicans Biofilm Formation with Precoating RNA-aptamer

F1000Research ◽

10.12688/f1000research.52266.2 ◽

2021 ◽

Vol 10 ◽

pp. 372

Author(s):

Boy Muchlis Bachtiar ◽

Basri A. Gani ◽

Astri Deviana ◽

Nastiti Rilo Utami ◽

Anissa Dien Andriyani ◽

...

Keyword(s):

Cigarette Smoke ◽

Growth Medium ◽

Biofilm Formation ◽

Morphological Changes ◽

Inhibitory Effect ◽

Yeast Cells ◽

Hypha Formation ◽

Microscopy Analysis ◽

Light Microscopy Analysis ◽

Biofilm Development

This study explores the influence of precoating aptamer (Ca-apt1) on C. albicans viability while the fungus was growing in the presence of exposing condensed cigarette smoke (CSC), prepared from clove (CCSC) and non-clove (NCSC) cigarettes, for 48 h. Using qPCR, we found that mRNA expression of adhesion-associated genes (ALS3 and HWP1) was impaired by precoating C. albicans yeast cells with the aptamer. Conversely, the gene transcription was upregulated when aptamer-uncoated yeast was pre-treated with either CSC. In addition, by analysing the result of MTT ([3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] assay, we found that the presence of added CCSC or NCSC in growth medium for 48 h was significantly enhanced C. albicans biofilm development. However, the presence of precoated aptamer was significantly impaired biofilm development accelerated by the NCSC. The inhibitory effect of the Ca-apt1 was not dependent on the precoated aptamer (1 and 10%). Interestingly, we noted that the enhancer effect of treated CCSC was no longer effective when the yeast had been precoated with 10% aptamer tested. Additionally, light microscopy analysis revealed that precoating aptamer alleviates morphological changes of C. albicans (from yeast to hypha formation) that are enhanced by adding CCSC or NCSC in the growth medium. In conclusion, these results suggest that administration on Ca-ap1 exhibits a significant protective effect on CSC-induced biofilm formation by C. albicans.

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Extracellular vesicles regulate yeast growth, biofilm formation, and yeast-to-hypha differentiation in Candida albicans

10.1101/2021.01.21.427696 ◽

2021 ◽

Author(s):

Leandro Honorato ◽

Joana Feital Demetrio ◽

Cameron C. Ellis ◽

Alicia Piffer ◽

Yan Pereira ◽

...

Keyword(s):

Candida Albicans ◽

Extracellular Vesicles ◽

Biofilm Formation ◽

Galleria Mellonella ◽

Morphological Changes ◽

Inhibitory Effect ◽

Yeast Cells ◽

Mass Spectrometry Analysis ◽

Yeast Growth

AbstractThe ability to undergo morphological changes during adaptation to distinct environments is exploited by Candida albicans and has a direct impact on virulence. In this study, we investigated the influence of fungal extracellular vesicles (EVs) during yeast growth, biofilm formation, and morphogenesis in C. albicans. Addition of C. albicans EVs (Ca EVs) to the culture medium positively affected yeast growth. Using crystal violet staining and scanning electron microscopy (SEM), we demonstrated that Ca EVs inhibited biofilm formation by C. albicans in vitro. By time-lapse microscopy and SEM, we showed that Ca EV-treatment stops filamentation promoting pseudohyphae formation with multiple sites for yeast budding. The ability of Ca EVs to regulate dimorphism was further compared to EVs isolated from different C. albicans strains, Saccharomyces cerevisiae, and Histoplasma capsulatum. Ca EVs from distinct strains robustly inhibited yeast-to-hyphae differentiation with morphological changes occurring in less than 4 hours. A minor inhibitory effect was promoted by EVs from S. cerevisiae and H. capsulatum only after 24 hours of incubation. The inhibitory effect of Ca EVs was promoted by a combination of lipid compounds identified by gas chromatography-tandem mass spectrometry analysis as sesquiterpenes, diterpenes, and fatty acids. Remarkably, Ca EVs were also able to reverse filamentation, transforming hyphal growth to yeast forms. Transcriptomic analysis demonstrated that treatment with Ca EVs modified the expression of more than 300 genes. The most effectively upregulated pathways were related to DNA metabolism. The downregulated genes were mostly associated with extracellular and adhesion proteins. Finally, yeast cells treated with Ca EVs for 24 hours lost their agar invasive ability and were avirulent when inoculated in Galleria mellonella larvae. In summary, our results indicate that fungal EVs can profoundly modify C. albicans growth and regulate yeast-to-hypha differentiation inhibiting biofilm formation and virulence.

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Comparative biofilm assays using Enterococcus faecalis OG1RF identify new determinants of biofilm formation

10.1101/2021.02.24.432758 ◽

2021 ◽

Author(s):

Julia L. E. Willett ◽

Jennifer L. Dale ◽

Lucy M. Kwiatkowski ◽

Jennifer L. Powers ◽

Michelle L. Korir ◽

...

Keyword(s):

Enterococcus Faecalis ◽

Growth Medium ◽

Biofilm Formation ◽

Time Course ◽

Opportunistic Pathogen ◽

Genetic Determinants ◽

Experimental Conditions ◽

Biofilm Reactors ◽

Biofilm Architecture ◽

Biofilm Development

AbstractEnterococcus faecalis is a common commensal organism and a prolific nosocomial pathogen that causes biofilm-associated infections. Numerous E. faecalis OG1RF genes required for biofilm formation have been identified, but few studies have compared genetic determinants of biofilm formation and biofilm morphology across multiple conditions. Here, we cultured transposon (Tn) libraries in CDC biofilm reactors in two different media and used Tn sequencing (TnSeq) to identify core and accessory biofilm determinants, including many genes that are poorly characterized or annotated as hypothetical. Multiple secondary assays (96-well plates, submerged Aclar, and MultiRep biofilm reactors) were used to validate phenotypes of new biofilm determinants. We quantified biofilm cells and used fluorescence microscopy to visualize biofilms formed by 6 Tn mutants identified using TnSeq and found that disrupting these genes (OG1RF_10350, prsA, tig, OG1RF_10576, OG1RF_11288, and OG1RF_11456) leads to significant time- and medium-dependent changes in biofilm architecture. Structural predictions revealed potential roles in cell wall homeostasis for OG1RF_10350 and OG1RF_11288 and signaling for OG1RF_11456. Additionally, we identified growth medium-specific hallmarks of OG1RF biofilm morphology. This study demonstrates how E. faecalis biofilm architecture is modulated by growth medium and experimental conditions, and identifies multiple new genetic determinants of biofilm formation.ImportanceE. faecalis is an opportunistic pathogen and a leading cause of hospital-acquired infections, in part due to its ability to form biofilms. A complete understanding of the genes required for E. faecalis biofilm formation as well as specific features of biofilm morphology related to nutrient availability and growth conditions is crucial for understanding how E. faecalis biofilm-associated infections develop and resist treatment in patients. We employed a comprehensive approach to analysis of biofilm determinants by combining TnSeq primary screens with secondary phenotypic validation using diverse biofilm assays. This enabled identification of numerous core (important under many conditions) and accessory (important under specific conditions) biofilm determinants in E. faecalis OG1RF. We found multiple genes whose disruption results in drastic changes to OG1RF biofilm morphology. These results expand our understanding of the genetic requirements for biofilm formation in E. faecalis that affect the time course of biofilm development as well as the response to specific nutritional conditions.

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Inhibitory effect of a natural phenolic compound, 3-p-trans-coumaroyl-2-hydroxyquinic acid against the attachment phase of biofilm formation of Staphylococcus aureus through targeting sortase A

RSC Advances ◽

10.1039/c9ra05883d ◽

2019 ◽

Vol 9 (56) ◽

pp. 32453-32461

Author(s):

Yan-Ping Wu ◽

Xiao-Yan Liu ◽

Jin-Rong Bai ◽

Hong-Chen Xie ◽

Si-Liang Ye ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Phenolic Compound ◽

Biofilm Formation ◽

Inhibitory Effect ◽

Sortase A ◽

Initial Attachment ◽

Biofilm Development

3-p-trans-Coumaroyl-2-hydroxyquinic acid (CHQA), a natural phenolic compound, prevented Staphylococcus aureus biofilm formation due to the inhibition of the initial attachment stage of biofilm development by targeting sortase A.

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Regulation of type 1 fimbriae synthesis and biofilm formation by the transcriptional regulator LrhA of Escherichia coli

Microbiology ◽

10.1099/mic.0.28098-0 ◽

2005 ◽

Vol 151 (10) ◽

pp. 3287-3298 ◽

Cited By ~ 76

Author(s):

Caroline Blumer ◽

Alexandra Kleefeld ◽

Daniela Lehnen ◽

Margit Heintz ◽

Ulrich Dobrindt ◽

...

Keyword(s):

Escherichia Coli ◽

Biofilm Formation ◽

Invertible Element ◽

Transcriptional Regulator ◽

Host Cells ◽

Yeast Cells ◽

Type 1 Fimbriae ◽

Biofilm Development

Type 1 fimbriae of Escherichia coli facilitate attachment to the host mucosa and promote biofilm formation on abiotic surfaces. The transcriptional regulator LrhA, which is known as a repressor of flagellar, motility and chemotaxis genes, regulates biofilm formation and expression of type 1 fimbriae. Whole-genome expression profiling revealed that inactivation of lrhA results in an increased expression of structural components of type 1 fimbriae. In vitro, LrhA bound to the promoter regions of the two fim recombinases (FimB and FimE) that catalyse the inversion of the fimA promoter, and to the invertible element itself. Translational lacZ fusions with these genes and quantification of fimE transcript levels by real-time PCR showed that LrhA influences type 1 fimbrial phase variation, primarily via activation of FimE, which is required for the ON-to-OFF transition of the fim switch. Enhanced type 1 fimbrial expression as a result of lrhA disruption was confirmed by mannose-sensitive agglutination of yeast cells. Biofilm formation was stimulated by lrhA inactivation and completely suppressed upon LrhA overproduction. The effects of LrhA on biofilm formation were exerted via the changed levels of surface molecules, most probably both flagella and type 1 fimbriae. Together, the data show a role for LrhA as a repressor of type 1 fimbrial expression, and thus as a regulator of the initial stages of biofilm development and, presumably, bacterial adherence to epithelial host cells also.

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Farnesol inhibits planktonic cells and antifungal-tolerant biofilms of Trichosporon asahii and Trichosporon inkin

Medical Mycology ◽

10.1093/mmy/myy160 ◽

2019 ◽

Vol 57 (8) ◽

pp. 1038-1045 ◽

Cited By ~ 5

Author(s):

Rossana de Aguiar Cordeiro ◽

Lívia Maria Galdino Pereira ◽

José Kleybson de Sousa ◽

Rosana Serpa ◽

Ana Raquel Colares Andrade ◽

...

Keyword(s):

Biofilm Formation ◽

Laser Scanning ◽

Inhibitory Effect ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Planktonic Cells ◽

Trichosporon Asahii ◽

Trichosporon Species ◽

Biofilm Development ◽

Systemic Infections

Abstract Trichosporon species have been considered important agents of opportunistic systemic infections, mainly among immunocompromised patients. Infections by Trichosporon spp. are generally associated with biofilm formation in invasive medical devices. These communities are resistant to therapeutic antifungals, and therefore the search for anti-biofilm molecules is necessary. This study evaluated the inhibitory effect of farnesol against planktonic and sessile cells of clinical Trichosporon asahii (n = 3) andTrichosporon inkin (n = 7) strains. Biofilms were evaluated during adhesion, development stages and after maturation for metabolic activity, biomass and protease activity, as well as regarding morphology and ultrastructure by optical microscopy, confocal laser scanning microscopy, and scanning electron microscopy. Farnesol inhibited Trichosporon planktonic growth by 80% at concentrations ranging from 600 to 1200 μM for T. asahii and from 75 to 600 μM for T. inkin. Farnesol was able to reduce cell adhesion by 80% at 300 μM for T. asahii and T. inkin at 600 μM, while biofilm development of both species was inhibited by 80% at concentration of 150 μM, altering their structure. After biofilm maturation, farnesol decreased T. asahii biofilm formation by 50% at 600 μM concentration and T. inkin formation at 300 μM. Farnesol inhibited gradual filamentation in a concentration range between 600 and 1200 μM. Farnesol caused reduction of filament structures of Trichosporon spp. at every stage of biofilm development analyzed. These data show the potential of farnesol as an anti-biofilm molecule.

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Polyphenols Inhibit Candida albicans and Streptococcus mutans Biofilm Formation

Dentistry Journal ◽

10.3390/dj7020042 ◽

2019 ◽

Vol 7 (2) ◽

pp. 42 ◽

Cited By ~ 7

Author(s):

Yosi Farkash ◽

Mark Feldman ◽

Isaac Ginsburg ◽

Doron Steinberg ◽

Miriam Shalish

Keyword(s):

Candida Albicans ◽

Streptococcus Mutans ◽

Biofilm Formation ◽

Inhibitory Effect ◽

Confocal Scanning Laser Microscopy ◽

Total Biomass ◽

Biofilm Inhibition ◽

Confocal Scanning ◽

Confocal Scanning Laser ◽

Biofilm Development

Background: Streptococcus mutans (S. mutans) and Candida albicans (C. albicans) are two major contributors to dental caries. They have a symbiotic relationship, allowing them to create an enhanced biofilm. Our goal was to examine whether two natural polyphenols (Padma hepaten (PH) and a polyphenol extraction from green tea (PPFGT)) could inhibit the caries-inducing properties of S. mutans and C. albicans. Methods: Co-species biofilms of S. mutans and C. albicans were grown in the presence of PH and PPFGT. Biofilm formation was tested spectrophotometrically. Exopolysaccharides (EPS) secretion was quantified using confocal scanning laser microscopy. Biofilm development was also tested on orthodontic surfaces (Essix) to assess biofilm inhibition ability on such an orthodontic appliance. Results: PPFGT and PH dose-dependently inhibited biofilm formation without affecting the planktonic growth. We found a significant reduction in biofilm total biomass using 0.625 mg/mL PPFGT and 0.16 mg/mL PH. A concentration of 0.31 mg/mL PPFGT and 0.16 mg/mL PH inhibited the total cell growth by 54% and EPS secretion by 81%. A reduction in biofilm formation and EPS secretion was also observed on orthodontic PVC surfaces. Conclusions: The polyphenolic extractions PPFGT and PH have an inhibitory effect on S. mutans and C. albicans biofilm formation and EPS secretion.

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Effect of chitosan nanoparticles on quorum sensing-controlled virulence factors and expression of LasI and RhlI genes among Pseudomonas aeruginosa clinical isolates

AIMS Microbiology ◽

10.3934/microbiol.2021025 ◽

2021 ◽

Vol 7 (4) ◽

pp. 415-430

Author(s):

Rana Abdel Fattah Abdel Fattah ◽

◽

Fatma El zaharaa Youssef Fathy ◽

Tahany Abdel Hamed Mohamed ◽

Marwa Shabban Elsayed

Keyword(s):

Pseudomonas Aeruginosa ◽

Quorum Sensing ◽

Virulence Factors ◽

Biofilm Formation ◽

Chitosan Nanoparticles ◽

Inhibitory Effect ◽

Biofilm Inhibition ◽

Expression Levels ◽

The Mean ◽

Biofilm Development

<abstract> <p>Antibiotic-resistant strains of <italic>Pseudomonas aeruginosa (P. aeruginosa</italic>) pose a major threat for healthcare-associated and community-acquired infections. <italic>P. aeruginosa</italic> is recognized as an opportunistic pathogen using quorum sensing (QS) system to regulate the expression of virulence factors and biofilm development. Thus, meddling with the QS system would give alternate methods of controlling the pathogenicity. This study aimed to assess the inhibitory impact of chitosan nanoparticles (CS-NPs) on <italic>P. aeruginosa</italic> virulence factors regulated by QS (e.g., motility and biofilm formation) and <italic>LasI</italic> and <italic>RhlI</italic> gene expression. Minimum inhibitory concentration (MIC) of CS-NPs against 30 isolates of <italic>P. aeruginosa</italic> was determined. The CS-NPs at sub-MIC were utilized to assess their inhibitory effect on motility, biofilm formation, and the expression levels of <italic>LasI</italic> and <italic>RhlI</italic> genes. CS-NPs remarkably inhibited the tested virulence factors as compared to the controls grown without the nanoparticles. The mean (±SD) diameter of swimming motility was decreased from 3.93 (±1.5) to 1.63 (±1.02) cm, and the mean of the swarming motility was reduced from 3.5 (±1.6) to 1.9 (±1.07) cm. All isolates became non-biofilm producers, and the mean percentage rate of biofilm inhibition was 84.95% (±6.18). Quantitative real-time PCR affirmed the opposition of QS activity by lowering the expression levels of <italic>LasI</italic> and <italic>RhlI</italic> genes; the expression level was decreased by 90- and 100-folds, respectively. In conclusion, the application of CS-NPs reduces the virulence factors significantly at both genotypic and phenotypic levels. These promising results can breathe hope in the fight against resistant <italic>P. aeruginosa</italic> by repressing its QS-regulated virulence factors.</p> </abstract>

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Production of Tyrosol by Candida albicans Biofilms and Its Role in Quorum Sensing and Biofilm Development

Eukaryotic Cell ◽

10.1128/ec.00219-06 ◽

2006 ◽

Vol 5 (10) ◽

pp. 1770-1779 ◽

Cited By ~ 151

Author(s):

Mohammed A. S. Alem ◽

Mohammed D. Y. Oteef ◽

T. Hugh Flowers ◽

L. Julia Douglas

Keyword(s):

Candida Albicans ◽

Quorum Sensing ◽

Biofilm Formation ◽

Dry Weight ◽

Cell Types ◽

Tube Formation ◽

Yeast Cells ◽

Planktonic Cells ◽

Biofilm Development ◽

Candida Albicans Biofilms

ABSTRACT Tyrosol and farnesol are quorum-sensing molecules produced by Candida albicans which accelerate and block, respectively, the morphological transition from yeasts to hyphae. In this study, we have investigated the secretion of tyrosol by C. albicans and explored its likely role in biofilm development. Both planktonic (suspended) cells and biofilms of four C. albicans strains, including three mutants with defined defects in the Efg 1 and Cph 1 morphogenetic signaling pathways, synthesized extracellular tyrosol during growth at 37°C. There was a correlation between tyrosol production and biomass for both cell types. However, biofilm cells secreted at least 50% more tyrosol than did planktonic cells when tyrosol production was related to cell dry weight. The addition of exogenous farnesol to a wild-type strain inhibited biofilm formation by up to 33% after 48 h. Exogenous tyrosol appeared to have no effect, but scanning electron microscopy revealed that tyrosol stimulated hypha production during the early stages (1 to 6 h) of biofilm development. Experiments involving the simultaneous addition of tyrosol and farnesol at different concentrations suggested that the action of farnesol was dominant, and 48-h biofilms formed in the presence of both compounds consisted almost entirely of yeast cells. When biofilm supernatants were tested for their abilities to inhibit or enhance germ tube formation by planktonic cells, the results indicated that tyrosol activity exceeds that of farnesol after 14 h, but not after 24 h, and that farnesol activity increases significantly during the later stages (48 to 72 h) of biofilm development. Overall, our results support the conclusion that tyrosol acts as a quorum-sensing molecule for biofilms as well as for planktonic cells and that its action is most significant during the early and intermediate stages of biofilm formation.

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