Combined effect of lasioglossin LL-III derivative with azoles against Candida albicans virulence factors: biofilm formation, phospholipases, proteases and hemolytic activity

ABSTRACT Candida albicans has several virulence factors at its disposal, including yeast–hyphal transition associated with biofilm formation, phospholipases, proteases and hemolytic activity, all of which contribute to its pathogenesis. We used synthetic derivative LL-III/43 of antimicrobial peptide lasioglossin LL-III to enhance effect of azoles on attenuation of C. albicans virulence factors. LL-III/43 was able to inhibit initial adhesion or biofilm formation of C. albicans strains at 50 µM. Azoles, however, were ineffective at this concentration. Using fluorescently labeled LL-III/43, we observed that peptide covered C. albicans cells, partially penetrated through their membranes and then accumulated inside cells. LL-III/43 (25 µM) in combination with clotrimazole prevented biofilm formation already at 3.1 µM clotrimazole. Neither LL-III/43 nor azoles were able to significantly inhibit phospholipases, proteases, or hemolytic activity of C. albicans. LL-III/43 (25 µM) and clotrimazole (50 µM) in combination decreased production of these virulence factors, and it completely attenuated its hemolytic activity. Scanning electron microscopy showed that LL-III/43 (50 µM) prevented C. albicans biofilm formation on Ti-6Al-4 V alloy used in orthopedic surgeries and combination of LL-III/43 (25 µM) with clotrimazole (3.1 µM) prevented biofilm formation on urinary catheters. Therefore, mixture of LL-III/43 and clotrimazole is suitable candidate for future pharmaceutical research.

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APPLICATION OF SCANNING ELECTRONIC MICROSCOPY FOR THE PURPOSE OF STUDYING CANDIDA ALBICANS BIOFILM ON THE SURFACE OF BASIC PLASTICS OF REMOVABLE ORTHOPEDIC DESIGNS

Russian Clinical Laboratory Diagnostics ◽

10.18821/0869-2084-2019-64-5-308-313 ◽

2019 ◽

Vol 64 (5) ◽

pp. 308-313 ◽

Cited By ~ 1

Author(s):

M. G. Chesnokova ◽

V. A. Chesnokov ◽

A. Yu. Mironov

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Candida Albicans ◽

Electron Microscope ◽

Biofilm Formation ◽

Scanning Electronic Microscopy ◽

Electronic Microscopy ◽

Orthopedic Rehabilitation ◽

Candida Albicans Biofilms ◽

Scanning Electron

The most common pathology in the clinic of orthopedic dentistry is the presence of partial adentia in patients, manifested in the form of defects of dentition of various localization and length. Removable orthopedic structures in the oral cavity are a potential place for adhesion and colonization of microorganisms. The aim of the research was to study Candida albicans biofilms on the surface of base plastics of removable orthopedic structures using scanning electron microscopy. 175 cultures of C. albicans were isolated and identified from the oral mucosa of patients at various stages of orthopedic rehabilitation. When studying the surface of samples of plastics of hot and cold type polymerization and Candida biofilms using a JEOL JCM 5700 scanning electron microscope (JEOL, Japan), features of biofilm formation were established. An assessment of the nature of the manifestation of the hemagglutinating activity of clinical strains of Candida fungi in the hemagglutination test with human erythrocytes I (O), II (A) of the human and guinea pig blood groups was carried out. The total number of hemagglutinating strains was 37.14%, with the prevalence of the proportion of manna-resistant (MRHA) cultures - 23.43% of cases. Micrographs of the C. albicans yeast-like biofilm biofilm were obtained on the surface of hot and cold-type plastics in incubation dynamics. Scanning electron microscopy revealed the most pronounced changes in the surface of hot plastics of polymerization compared to cold plastics with long incubation of C. albicans, which characterize the loosening of plastics and the appearance of cracks on the surface, and the cracking of a yeast-like fungus biofilm was noted.

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Adhesion and biofilm formation of Candida albicans on native and Pluronic-treated polystyrene

Biofilms ◽

10.1017/s1479050505001687 ◽

2005 ◽

Vol 2 (1) ◽

pp. 63-71 ◽

Cited By ~ 21

Author(s):

K. E. Wesenberg-Ward ◽

B. J. Tyler ◽

J. T. Sears

Keyword(s):

Candida Albicans ◽

Biofilm Formation ◽

Pluronic F127 ◽

Artificial Organs ◽

Confocal Scanning Laser Microscopy ◽

Mechanical Trauma ◽

Biofilm Growth ◽

Prosthetic Devices ◽

Initial Adhesion ◽

Defined Culture

Candida albicans forms part of the normal human flora whose growth is usually restricted by the normal flora bacteria and the host's immune system. It is an opportunistic fungal pathogen that causes infections in immunocompromised individuals, mechanical trauma victims and iatrogenic patients. Candida albicans can ingress the human host by adhering to a plastic surface (i.e. prosthetic devices, catheters, artificial organs, etc.) that is subsequently implanted, and forms a protective biofilm that provides a continuous reservoir of yeast to be hematogenously dispersed. In order for the medical profession to battle device-related infections, initial adhesion and biofilm formation of C. albicans needs to be better understood. There has been some skepticism as to whether the initial adhesion events bear any relationship to subsequent biofilm formation. Thus, to better comprehend the relationship between the initial adhesion rates and growth rate and biofilm formation, these events were studied on two different, well-defined culture surfaces, native polystyrene and Pluronic F127-conditioned polystyrene. The adhesion studies determined that Pluronic F127 adsorption dramatically reduced the adhesion of C. albicans to polystyrene. The biofilm growth studies, analyzed by confocal scanning laser microscopy, revealed that Pluronic F127 decreased the biofilm surface coverage, cluster group size, thickness and the presence of hyphal elements over the untreated polystyrene. These findings indicate that the effect of a material's surface chemistry on the initial adhesion process has a direct influence on subsequent biofilm formation.

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Cajuputs candy impairs Candida albicans and Streptococcus mutans mixed biofilm formation in vitro

F1000Research ◽

10.12688/f1000research.20700.2 ◽

2020 ◽

Vol 8 ◽

pp. 1923

Author(s):

Siska Septiana ◽

Boy Muchlis Bachtiar ◽

Nancy Dewi Yuliana ◽

Christofora Hanny Wijaya

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Candida Albicans ◽

Streptococcus Mutans ◽

Biofilm Formation ◽

Microscopy Imaging ◽

Biofilm Inhibition ◽

Yeast Form ◽

Melaleuca Cajuputi ◽

Scanning Electron

Background: Cajuputs candy (CC), an Indonesian functional food, utilizes the bioactivity of Melaleuca cajuputi essential oil (MCEO) to maintain oral cavity health. Synergistic interaction between Candida albicans and Streptococcus mutans is a crucial step in the pathogenesis of early childhood caries. Our recent study revealed several alternative MCEOs as the main flavors in CC. The capacity of CC to interfere with the fungus-bacterium relationship remains unknown. This study aimed to evaluate CC efficacy to impair biofilm formation by these dual cariogenic microbes. Methods: The inhibition capacity of CC against mixed-biofilm comprising C. albicans and S. mutans was assessed by quantitative (crystal violet assay, tetrazolium salt [MTT] assay, colony forming unit/mL counting, biofilm-related gene expression) and qualitative analysis (light microscopy and scanning electron microscopy). Result: Both biofilm-biomass and viable cells were significantly reduced in the presence of CC. Scanning electron microscopy imaging confirmed this inhibition capacity, demonstrating morphology alteration of C. albicans, along with reduced microcolonies of S. mutans in the biofilm mass. This finding was related to the transcription level of selected biofilm-associated genes, expressed either by C. albicans or S. mutans. Based on qPCR results, CC could interfere with the transition of C. albicans yeast form to the hyphal form, while it suppressed insoluble glucan production by S. mutans. G2 derived from Mojokerto MCEO showed the greatest inhibition activity on the relationship between these cross-kingdom oral microorganisms (p < 0.05). Conclusion: In general, all CC formulas showed biofilm inhibition capacity. Candy derived from Mojokerto MCEO showed the greatest capacity to maintain the yeast form of C. albicans and to inhibit extracellular polysaccharide production by S. mutans. Therefore, the development of dual-species biofilms can be impaired effectively by the CC tested.

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First report of macroscopic biofilm formation caused by Candida albicans on silver hydrogel–coated urinary catheters

American Journal of Infection Control ◽

10.1016/j.ajic.2016.03.042 ◽

2016 ◽

Vol 44 (10) ◽

pp. 1174-1175 ◽

Cited By ~ 2

Author(s):

Mehmet Burak Selek ◽

Tuğba Kula Atik ◽

Bayhan Bektöre ◽

Bülent Atik ◽

Serkan Demir ◽

...

Keyword(s):

Candida Albicans ◽

Biofilm Formation ◽

Urinary Catheters ◽

First Report

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E1210, a New Broad-Spectrum Antifungal, Suppresses Candida albicans Hyphal Growth through Inhibition of Glycosylphosphatidylinositol Biosynthesis

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00731-11 ◽

2011 ◽

Vol 56 (2) ◽

pp. 960-971 ◽

Cited By ~ 77

Author(s):

Nao-aki Watanabe ◽

Mamiko Miyazaki ◽

Takaaki Horii ◽

Koji Sagane ◽

Kappei Tsukahara ◽

...

Keyword(s):

Candida Albicans ◽

Virulence Factors ◽

Inhibitory Activity ◽

Biofilm Formation ◽

Protein A ◽

Hyphal Growth ◽

Tube Formation ◽

Protein Maturation ◽

Content Type ◽

Protein Levels

ABSTRACTContinued research toward the development of new antifungals that act via inhibition of glycosylphosphatidylinositol (GPI) biosynthesis led to the design of E1210. In this study, we assessed the selectivity of the inhibitory activity of E1210 againstCandida albicansGWT1(Orf19.6884) protein,Aspergillus fumigatusGWT1(AFUA_1G14870) protein, and humanPIG-Wprotein, which can catalyze the inositol acylation of GPI early in the GPI biosynthesis pathway, and then we assessed the effects of E1210 on keyC. albicansvirulence factors. E1210 inhibited the inositol acylation activity ofC. albicansGwt1p andA. fumigatusGwt1p with 50% inhibitory concentrations (IC50s) of 0.3 to 0.6 μM but had no inhibitory activity against human Pig-Wp even at concentrations as high as 100 μM. To confirm the inhibition of fungal GPI biosynthesis, expression ofALS1protein, a GPI-anchored protein, on the surfaces ofC. albicanscells treated with E1210 was studied and shown to be significantly lower than that on untreated cells. However, theALS1protein levels in the crude extract and theRHO1protein levels on the cell surface were found to be almost the same. Furthermore, E1210 inhibited germ tube formation, adherence to polystyrene surfaces, and biofilm formation ofC. albicansat concentrations above its MIC. These results suggested that E1210 selectively inhibited inositol acylation of fungus-specific GPI which would be catalyzed by Gwt1p, leading to the inhibition of GPI-anchored protein maturation, and also that E1210 suppressed the expression of some important virulence factors ofC. albicans, through its GPI biosynthesis inhibition.

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Disarming Fungal Pathogens:Bacillus safensisInhibits Virulence Factor Production and Biofilm Formation byCryptococcus neoformansandCandida albicans

mBio ◽

10.1128/mbio.01537-17 ◽

2017 ◽

Vol 8 (5) ◽

Cited By ~ 24

Author(s):

François L. Mayer ◽

James W. Kronstad

Keyword(s):

Cell Wall ◽

Candida Albicans ◽

Cryptococcus Neoformans ◽

Virulence Factors ◽

Virulence Factor ◽

Biofilm Formation ◽

Fungal Pathogens ◽

Soil Bacterium ◽

Fungal Cell ◽

Virulence Factor Production

ABSTRACTBacteria interact with each other in nature and often compete for limited nutrient and space resources. However, it is largely unknown whether and how bacteria also interact with human fungal pathogens naturally found in the environment. Here, we identified a soil bacterium,Bacillus safensis, which potently blocked several keyCryptococcus neoformansvirulence factors, including formation of the antioxidant pigment melanin and production of the antiphagocytic polysaccharide capsule. The bacterium also inhibitedde novocryptococcal biofilm formation but had only modest inhibitory effects on already formed biofilms or planktonic cell growth. The inhibition of fungal melanization was dependent on direct cell contact and live bacteria.B. safensisalso had anti-virulence factor activity against another major human-associated fungal pathogen,Candida albicans. Specifically, dual-species interaction studies revealed that the bacterium strongly inhibitedC. albicansfilamentation and biofilm formation. In particular,B. safensisphysically attached to and degraded candidal filaments. Through genetic and phenotypic analyses, we demonstrated that bacterial chitinase activity against fungal cell wall chitin is a factor contributing to the antipathogen effect ofB. safensis.IMPORTANCEPathogenic fungi are estimated to contribute to as many human deaths as tuberculosis or malaria. Two of the most common fungal pathogens,Cryptococcus neoformansandCandida albicans, account for up to 1.4 million infections per year with very high mortality rates. Few antifungal drugs are available for treatment, and development of novel therapies is complicated by the need for pathogen-specific targets. Therefore, there is an urgent need to identify novel drug targets and new drugs. Pathogens use virulence factors during infection, and it has recently been proposed that targeting these factors instead of the pathogen itself may represent a new approach to develop antimicrobials. Here, we identified a soil bacterium that specifically blocked virulence factor production and biofilm formation byC. neoformansandC. albicans. We demonstrate that the bacterial antipathogen mechanism is based in part on targeting the fungal cell wall, a structure not found in human cells.

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Cajuputs candy impairs Candida albicans and Streptococcus mutans mixed biofilm formation in vitro

F1000Research ◽

10.12688/f1000research.20700.1 ◽

2019 ◽

Vol 8 ◽

pp. 1923 ◽

Cited By ~ 1

Author(s):

Siska Septiana ◽

Boy Muchlis Bachtiar ◽

Nancy Dewi Yuliana ◽

Christofora Hanny Wijaya

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Candida Albicans ◽

Streptococcus Mutans ◽

Biofilm Formation ◽

Tetrazolium Salt ◽

Microscopy Imaging ◽

Biofilm Inhibition ◽

Melaleuca Cajuputi ◽

Scanning Electron

Background: Cajuputs candy (CC), an Indonesian functional food, utilizes the bioactivity of Melaleuca cajuputi essential oil (MCEO) to maintain oral cavity health. Synergistic interaction between Candida albicans and Streptococcus mutans is a crucial step in the pathogenesis of early childhood caries. Our recent study revealed several alternative MCEOs as the main flavors in CC. The capacity of CC to interfere with the fungus-bacterium relationship remains unknown. This study aimed to evaluate CC efficacy to impair biofilm formation by these dual cariogenic microbes. Methods: The inhibition capacity of CC against mixed-biofilm comprising C. albicans and S. mutans was assessed by quantitative (crystal violet assay, tetrazolium salt [MTT] assay, colony forming unit/mL counting, biofilm-related gene expression) and qualitative analysis (light microscopy and scanning electron microscopy). Result: Both biofilm-biomass and viable cells were significantly reduced in the presence of CC. Scanning electron microscopy imaging confirmed this inhibition capacity, demonstrating morphology alteration of C. albicans, along with reduced microcolonies of S. mutans in the biofilm mass. This finding was related to the transcription level of selected biofilm-associated genes, expressed either by C. albicans or S. mutans. Based on qPCR results, CC could interfere with the transition of C. albicans yeast form to the hyphal form, while it suppressed insoluble glucan production by S. mutans. G2 derived from Mojokerto MCEO showed the greatest inhibition activity on the relationship between these cross-kingdom oral microorganisms (p < 0.05). Conclusion: In general, all CC formulas showed biofilm inhibition capacity. Candy derived from Mojokerto MCEO showed the greatest capacity to maintain the commensal form of C. albicans and to inhibit extracellular polysaccharide production by S. mutans. Therefore, the development of dual-species biofilms can be impaired effectively by the CC tested.

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Biofabricated Silver Nanoparticles Synergistically Activate Amphotericin B Against Mature Biofilm Forms of Candida Albicans

The Open Nanomedicine Journal ◽

10.2174/1875933501704010001 ◽

2017 ◽

Vol 4 (1) ◽

pp. 1-16 ◽

Cited By ~ 4

Author(s):

Shivkrupa D. Halbandge ◽

Supriya P. Mortale ◽

Sankunny Mohan Karuppayil

Keyword(s):

Silver Nanoparticles ◽

Candida Albicans ◽

Amphotericin B ◽

Hemolytic Activity ◽

Biofilm Formation ◽

Inhibitory Concentration ◽

Fungal Infections ◽

Fractional Inhibitory Concentration ◽

Planktonic Growth ◽

Concentration Indices

Background: Biofilm formation by Candida albicans is a significant clinical challenge. Fungal biofilms are resistant to most of the currently available antifungal agents. Amphotericin-B (AmB) is an antifungal agent used for the treatment of systematic fungal infections but it is well known for its toxicities and side-effects. Novel approaches are needed to treat these infections that can reduce its toxicities. Objectives: Current study aims to evaluate the efficacy of silver nanoparticles (SNPs) alone and in combination with AmB against growth and biofilm formation in C. albicans. Methods: Combinations of SNP-AmB were tested against planktonic growth and biofilm formation in vitro. Micro broth dilution method was used to study planktonic growth and biofilm formation. The fractional inhibitory concentration indices (FICI) were calculated by using a checkerboard format. Biofilm formation was analyzed by using XTT-metabolic assay. Results: MIC of AmB for developing biofilm was lowered by 16 fold in combination with SNPs. The calculated fractional inhibitory concentration indices were 0.1875 suggesting that this interaction is synergistic. Similarly, the mature biofilms were significantly prevented by SNPs-AmB combination. This interaction was synergistic. Furthermore, interaction between SNPs and AmB against planktonic growth was additive. Hemolytic activity assay was carried out on these drugs and combinations. Drug required for inhibition alone as well as in combination did not exhibit hemolytic activity. Conclusion: The combinations with SNPs lead to decreases in the dosage of AmB required for anti-Candida activity. SNPs-AmB combination could be an effective strategy against biofilm formed by C. albicans.

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In Vitro Study on the Adhesion and Colonization of Candida Albicans on Metal and Acrylic Piercings

Acta Medica Bulgarica ◽

10.1515/amb-2016-0001 ◽

2016 ◽

Vol 43 (1) ◽

pp. 5-13

Author(s):

N. Stamenov ◽

G. Tomov ◽

Z. Denkova ◽

I. Dobrev

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Candida Albicans ◽

Metal Surface ◽

Biofilm Formation ◽

In Vitro Study ◽

Electrostatic Forces ◽

Plastic Surface ◽

Scanning Electron

SummaryOral/perioral piercing may provide an ideal environment for adhesion and colonization of microorganisms. The aim of this study is to perform an “in vitro” research on the capabilities of adhesion of Candida albicans on oral piercings made of plastic and metal. Acrylic and metal piercings were incubated with Candida albicans and then were observed using scanning electron microscopy under different magnifications. A lot of irregularities and roughness were observed on the surface of the plastic piercing unlike the surface of the metal one, which is not so rough. Nevertheless, the number of Candida albicans colonies was considerably larger on the scanned metal surface in comparison to the plastic surface. In vitro the metal surface of the piercing creates better environment for the adhesion and colonization of microorganisms than the acrylic. This could be attributed to the electrostatic forces that most likely attract Candida albicans to the metal piercing in the early stages of biofilm formation.

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