In vitro antifungal effect of a plant-based product, CIN-102, on antifungal resistant filamentous fungi and their biofilms

Introduction. The increase of invasive fungal infections (IFIs) and associated treatment failure in populations at risk is driving us to look for new treatments. Hypothesis. The CIN-102 compound, derived from cinnamon essential oil, could be a new antifungal class with an activity, in particular, on strains resistant to current antifungals but also on biofilms, a factor of virulence and resistance of fungi. Aim. The aim of this study is to show the activity of CIN-102 on various strains resistant to current antifungals, on the biofilm and to determine the possibility of resistance induced with this compound. Methodology. We studied the MIC of CIN-102 and of current antifungals (voriconazole and amphotericin B) using CLSI techniques against eight different strains of three genera of filamentous fungi involved in IFIs and having resistance phenotypes to current antifungals. We also determined their effects on biofilm formation, and the induced resistance by voriconazole (VRC) and CIN-102. Results. MIC values determined for CIN-102 were between 62.5 and 250 µg ml−1. We demonstrated the antifungal effect of CIN-102 on biofilm, and more particularly on its formation, with 100 % inhibition achieved for most of the strains. CIN-102 at a sub-inhibitory concentration in the medium did not induce resistance in our strains, even after 30 generations. Conclusions. In this study we show that CIN-102 is effective against resistant filamentous fungi and against biofilm formation. In addition, our strains did not acquire a resistance phenotype against CIN-102 over time, unlike with VRC. CIN-102 is therefore an interesting candidate for the treatment of IFIs, including in cases of therapeutic failure linked to resistance, although further studies on its efficacy, safety and mechanism of action are needed.

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Eap1p, an Adhesin That Mediates Candida albicans Biofilm Formation In Vitro and In Vivo

Eukaryotic Cell ◽

10.1128/ec.00049-07 ◽

2007 ◽

Vol 6 (6) ◽

pp. 931-939 ◽

Cited By ~ 96

Author(s):

Fang Li ◽

Michael J. Svarovsky ◽

Amy J. Karlsson ◽

Joel P. Wagner ◽

Karen Marchillo ◽

...

Keyword(s):

Candida Albicans ◽

Cell Adhesion ◽

Biofilm Formation ◽

Fungal Infections ◽

Gene Dosage ◽

Venous Catheter ◽

Flow Chamber ◽

Dependent Manner

ABSTRACT Candida albicans is the leading cause of systemic fungal infections in immunocompromised humans. The ability to form biofilms on surfaces in the host or on implanted medical devices enhances C. albicans virulence, leading to antimicrobial resistance and providing a reservoir for infection. Biofilm formation is a complex multicellular process consisting of cell adhesion, cell growth, morphogenic switching between yeast form and filamentous states, and quorum sensing. Here we describe the role of the C. albicans EAP1 gene, which encodes a glycosylphosphatidylinositol-anchored, glucan-cross-linked cell wall protein, in adhesion and biofilm formation in vitro and in vivo. Deleting EAP1 reduced cell adhesion to polystyrene and epithelial cells in a gene dosage-dependent manner. Furthermore, EAP1 expression was required for C. albicans biofilm formation in an in vitro parallel plate flow chamber model and in an in vivo rat central venous catheter model. EAP1 expression was upregulated in biofilm-associated cells in vitro and in vivo. Our results illustrate an association between Eap1p-mediated adhesion and biofilm formation in vitro and in vivo.

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Symbiotic NCR Peptide Fragments Affect the Viability, Morphology and Biofilm Formation of Candida Species

International Journal of Molecular Sciences ◽

10.3390/ijms22073666 ◽

2021 ◽

Vol 22 (7) ◽

pp. 3666

Author(s):

Bettina Szerencsés ◽

Attila Gácser ◽

Gabriella Endre ◽

Ildikó Domonkos ◽

Hilda Tiricz ◽

...

Keyword(s):

Candida Species ◽

Biofilm Formation ◽

Therapeutic Potential ◽

Fungal Infections ◽

Combined Treatment ◽

Antimicrobial Activities ◽

Dilution Method ◽

Model Legume ◽

Anticandidal Activity

The increasing rate of fungal infections causes global problems not only in human healthcare but agriculture as well. To combat fungal pathogens limited numbers of antifungal agents are available therefore alternative drugs are needed. Antimicrobial peptides are potent candidates because of their broad activity spectrum and their diverse mode of actions. The model legume Medicago truncatula produces >700 nodule specific cysteine-rich (NCR) peptides in symbiosis and many of them have in vitro antimicrobial activities without considerable toxicity on human cells. In this work we demonstrate the anticandidal activity of the NCR335 and NCR169 peptide derivatives against five Candida species by using the micro-dilution method, measuring inhibition of biofilm formation with the XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) assay, and assessing the morphological change of dimorphic Candida species by microscopy. We show that both the N- and C-terminal regions of NCR335 possess anticandidal activity as well as the C-terminal sequence of NCR169. The active peptides inhibit biofilm formation and the yeast-hypha transformation. Combined treatment of C. auris with peptides and fluconazole revealed synergistic interactions and reduced 2-8-fold the minimal inhibitory concentrations. Our results demonstrate that shortening NCR peptides can even enhance and broaden their anticandidal activity and therapeutic potential.

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Biofilm Formation by Pneumocystis spp.

Eukaryotic Cell ◽

10.1128/ec.00202-08 ◽

2008 ◽

Vol 8 (2) ◽

pp. 197-206 ◽

Cited By ~ 60

Author(s):

Melanie T. Cushion ◽

Margaret S. Collins ◽

Michael J. Linke

Keyword(s):

Biofilm Formation ◽

Fungal Infections ◽

Morphological Changes ◽

Continuous Cultivation ◽

Life Cycles ◽

Mammalian Host ◽

Intense Staining ◽

In Vitro System ◽

Major Surface

ABSTRACT Pneumocystis spp. can cause a lethal pneumonia in hosts with debilitated immune systems. The manner in which these fungal infections spread throughout the lung, the life cycles of the organisms, and their strategies used for survival within the mammalian host are largely unknown, due in part to the lack of a continuous cultivation method. Biofilm formation is one strategy used by microbes for protection against environmental assaults, for communication and differentiation, and as foci for dissemination. We posited that the attachment and growth of Pneumocystis within the lung alveoli is akin to biofilm formation. An in vitro system comprised of insert wells suspended in multiwell plates containing supplemented RPMI 1640 medium supported biofilm formation by P. carinii (from rat) and P. murina (from mouse).Dramatic morphological changes accompanied the transition to a biofilm. Cyst and trophic forms became highly refractile and produced branching formations that anastomosed into large macroscopic clusters that spread across the insert. Confocal microscopy revealed stacking of viable organisms enmeshed in concanavalin A-staining extracellular matrix. Biofilms matured over a 3-week time period and could be passaged. These passaged organisms were able to cause infection in immunosuppressed rodents. Biofilm formation was inhibited by farnesol, a quorum-sensing molecule in Candida spp., suggesting that a similar communication system may be operational in the Pneumocystis biofilms. Intense staining with a monoclonal antibody to the major surface glycoproteins and an increase in (1,3)-β-d-glucan content suggest that these components contributed to the refractile properties. Identification of this biofilm process provides a tractable in vitro system that should fundamentally advance the study of Pneumocystis.

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Environment in the lung of cystic fibrosis patients stimulates the expression of biofilm phenotype in Mycobacterium abscessus

Journal of Medical Microbiology ◽

10.1099/jmm.0.001467 ◽

2022 ◽

Vol 71 (1) ◽

Author(s):

Bailey F. Keefe ◽

Luiz E. Bermudez

Keyword(s):

Cystic Fibrosis ◽

Host Cell ◽

Biofilm Formation ◽

Type Species ◽

Divalent Cations ◽

Mycobacterium Abscessus ◽

Content Type ◽

Link Type ◽

Populations At Risk

Introduction. Pulmonary infections caused by organisms of the Mycobacterium abscessus complex are increasingly prevalent in populations at risk, such as patients with cystic fibrosis, bronchiectasis and emphysema. Hypothesis. M. abscessus infection of the lung is not observed in immunocompetent individuals, which raises the possibility that the compromised lung environment is a suitable niche for the pathogen to thrive in due to the overproduction of mucus and high amounts of host cell lysis. Aim. Evaluate the ability of M. abscessus to form biofilm and grow utilizing in vitro conditions as seen in immunocompromised lungs of patients. Methodology. We compared biofilm formation and protein composition in the presence and absence of synthetic cystic fibrosis medium (SCFM) and evaluated the bacterial growth when exposed to human DNA. Results. M. abscessus is capable of forming biofilm in SCFM. By eliminating single components found in the medium, it became clear that magnesium works as a signal for the biofilm formation, and chelation of the divalent cations resulted in the suppression of biofilm formation. Investigation of the specific proteins expressed in the presence of SCFM and in the presence of SCFM lacking magnesium revealed many different proteins between the conditions. M. abscessus also exhibited growth in SCFM and in the presence of host cell DNA, although the mechanism of DNA utilization remains unclear. Conclusions. In vitro conditions mimicking the airways of patients with cystic fibrosis appear to facilitate M. abscessus establishment of infection, and elimination of magnesium from the environment may affect the ability of the pathogen to establish infection.

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Susceptibility of Dermatophytes to Thiabendazole Using CLSI Broth Macrodilution

ISRN Dermatology ◽

10.5402/2012/351842 ◽

2012 ◽

Vol 2012 ◽

pp. 1-3 ◽

Cited By ~ 3

Author(s):

Efrén Robledo-Leal ◽

Mariana Elizondo-Zertuche ◽

Gloria M. González

Keyword(s):

Antifungal Activity ◽

Filamentous Fungi ◽

Reference Method ◽

Strong Inhibitor ◽

Antifungal Effect ◽

Susceptibility Data ◽

Standardized Protocol ◽

Broth Macrodilution

Objective. To evaluate in vitro antifungal activity of thiabendazole against strains of dermatophytes using a reference method for filamentous fungi. Materials and Methods. Dermatophytes’ susceptibility to thiabendazole (TBZ) and fluconazole (FCZ) was evaluated using macrodilution method of protocol M38-A2 of the Clinical and Laboratory Standards Institute (CLSI). Results. MIC ranges of TBZ for all strains were narrower and/or smaller than those of FCZ. TBZ showed a significantly greater potency than FCZ () against all isolates. Discussion. Although there have been approaches to evaluate the antifungal activity of TBZ in human mycoses, no tests had been made with a standardized protocol. Susceptibility data resulted from this study shows that although TBZ is not a particularly strong inhibitor of dermatophytes, it displays a stable and constant effect against all isolates tested. Conclusion. Results show that TBZ is more effective against strains of dermatophytes than FCZ. We acknowledge the antifungal effect of TBZ against dermatophyte isolates.

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Anti-Trichophyton Effects of Curcumin: A Systematic Review

Anti-Infective Agents ◽

10.2174/2211352519666210202085849 ◽

2021 ◽

Vol 19 ◽

Author(s):

Pegah Shakib ◽

Ali Safar Maken Ali ◽

Ehsan Javanmard ◽

Omid Mardanshah ◽

Mohammad Ali Mohaghegh ◽

...

Keyword(s):

Systematic Review ◽

Web Of Science ◽

Fungal Infections ◽

Good Alternative ◽

Potent Effect ◽

Antifungal Effects ◽

Review Study ◽

Different Strains

Introduction: Today we see advances in the use of medicinal plants in the treatment of fungal infections. Curcumin has major antimicrobial, antifungal, antimutagenic, and anticancer activity. This systematic review study is designed to evaluate the effectiveness of curcuminon a dermatophyte fungus, Trichophyton spp. Methods: In this study, antifungal effects of curcumin on the different strains of Trichophytonwereevaluated. For this reason, several databases, including PubMed, Scopus, Web of Science, Embase and Google Scholar were searched systematically in years from1995 to 2020. Only articles with at least English abstracts were evaluated. The syntax was constructed with the combination of some key words along with specific tags used for each of the databases. Results: In this systematic review, studies showed that curcumin has a potent effect on the inhibition of Trichophyton growth. Of the 2,500 studies in the search step, only 11articleswere eligible for survey, 6 of which were on T.rubrum, 3 on T. mentagrophytes, 1 on both T. mentagrophyte and T. rubrum, and 1 on T. longifusus. Conclusion: Today, due to an increase in drug resistance to antifungal agents, plant extracts can be a good alternative in controlling fungal diseases. Curcumin and its compounds are effective in inhibiting or reducing Trichophyton infections in vitro and in vivo.

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Effect of Commonly Prescribed Liquid Medications on Streptococcus mutans Biofilm. An in vitro study

Journal of Clinical Pediatric Dentistry ◽

10.17796/1053-4628-41.2.141 ◽

2017 ◽

Vol 41 (2) ◽

pp. 141-146 ◽

Cited By ~ 4

Author(s):

Samantha A Clark ◽

LaQuia A Vinson ◽

George Eckert ◽

Richard L Gregory

Keyword(s):

Streptococcus Mutans ◽

Biofilm Formation ◽

Fungal Infections ◽

Sugar Content ◽

In Vitro Study ◽

Flat Bottom ◽

Microtiter Plates ◽

Oral Liquid ◽

High Concentrations

Objective: This study addressed the effect of pediatric liquid antibiotic medications on Streptococcus mutans UA159. These suspensions commonly contain sugars such as sucrose to make them more palatable for children. The study was designed to evaluate the effects of oral liquid antibiotics on Streptococcus mutans growth and biofilm formation. Study Design: A 24 hour culture of S. mutans was treated with various concentrations of liquid medications commonly prescribed to children for odontogenic or fungal infections– amoxicillin, penicillin VK, clindamycin, and nystatin. The study was conducted in sterile 96-well flat bottom microtiter plates. The minimum inhibitory and biofilm inhibitory concentrations (MIC/MBIC) of S. mutans were determined for each medication. S. mutans was cultured with and without the test drugs, the amount of total growth measured, the biofilms washed, fixed, and stained with crystal violet. The absorbance was determined to evaluate biofilm formation. Results: Higher concentrations of amoxicillin, penicillin VK and clindamycin had decreased biofilm and overall growth than the control. The MICs were 1:2,560 (1.95 ug/ml), 1:2,560 (1.95 ug/ml) and 1:40 (9.375 ug/ml), while the MBIC were 1:640 (7.8 ug/ml), 1:1,280 (3.9 ug/ml) and 1:20 (18.75 ug/ml), respectively. Lower concentrations provided increased biofilm and overall growth. Nystatin induced significantly more biofilm and overall growth than the control at all concentrations. Conclusion: At high concentrations, approximately at the levels expected to be present in the oral cavity of children, amoxicillin, penicillin, and clindamycin inhibited S. mutans biofilm and overall growth due to their antibiotic activity, while at lower concentrations the three antibiotics demonstrated an increase in biofilm and growth. The increase in S. mutans biofilm and overall growth is most likely attributed to the sugar content in the medications. Nystatin provided an increase in biofilm and growth at each concentration tested.

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ANTIMICROBIAL AND ANTIRADICAL PROPERTIES OF HAMMADA SCOPARIA (POMEL) ILJIN

African Journal of Traditional Complementary and Alternative Medicines ◽

10.21010/ajtcam.v16n2.1 ◽

2020 ◽

Vol 16 (2) ◽

pp. 1-14

Author(s):

Aziz Drioiche ◽

Nadia Benhlima ◽

Samira Kharchouf ◽

Fadoua EL MAKHOUKHI ◽

Smahane MEHANNED ◽

...

Keyword(s):

Fungal Infections ◽

Chemical Characterization ◽

Traditional Healers ◽

Bacterial Strains ◽

Traditional Use ◽

Antifungal Effect ◽

Aerial Parts ◽

Organic Extracts

Background: Hammada scoparia (Pomel) Iljin (HS), is commonly used by traditional healers in Morocco against microbial and fungal infections. We studied antimicrobial, antifungal and antiradical effects of organic extracts in vitro in order to confirm traditional utilization after phytochemical screening. Materials and methods: Aerial parts of HS have been extracted by hydro-distillation using Clevenger-type apparatus, and the chemical composition was realized by Gas Chromatography coupled with Mass Spectroscopy (GC/MS). The antioxidant activity has been evaluated using DPPH test, while the antimicrobial tests of HS extract were conducted on twenty-eight bacterial strains and antifungal on twelve fungal strains. Results: Chemical characterization of HS essential oils (EO) confirmed the presence of carvacrol (82,28%), p-cymene (2,52%), γ-terpinene (2,18%) and Z-caryophyllene (2,04%). Antimicrobial tests of HS extract showed a moderate antibacterial activity without antifungal effect. In addition, HS exhibited a very powerful antiradical activity with IC50 = 1,2 mg/ml compared to that of ascorbic acid (IC50 = 0,5 mg/mL) and butylated hydroxyanisole (0,8 mg/mL). Conclusion: To our knowledge, this is the first demonstration that HS directly inhibits the growth of microorganisms in vitro, and further validates its traditional use as an antiseptic by traditional Moroccan healers.

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S. aureus Colonization, Biofilm Production, and Phage Susceptibility in Peritoneal Dialysis Patients

Antibiotics ◽

10.3390/antibiotics9090582 ◽

2020 ◽

Vol 9 (9) ◽

pp. 582

Author(s):

Karlis Racenis ◽

Juta Kroica ◽

Dace Rezevska ◽

Lauris Avotins ◽

Edgars Skuditis ◽

...

Keyword(s):

Peritoneal Dialysis ◽

Biofilm Formation ◽

Bacterial Resistance ◽

In Vitro Susceptibility ◽

Phenotypic Differences ◽

Biofilm Production ◽

Adaptation Procedure ◽

Different Strains ◽

Vitro Effect

Peritonitis caused by Staphylococcusaureus is of major importance in peritoneal dialysis (PD) patients due to its great virulence profile and biofilm formation ability. Bacteriophages are a potential tool to treat peritonitis resulting from biofilm-associated infections. We screened S. aureus colonization in 71 PD patients from the nasal cavity, groin, and PD exit-site regions and analyzed clinical outcomes in these patients. We performed biofilm-formation testing of different strains and compared the isolates of one patient to detect phenotypic differences in S. aureus. Phage cocktails were used to detect S. aureus in vitro susceptibility. An adaptation procedure was performed in cases of bacterial resistance. Around 30% of PD patients (n = 21) were found to be S. aureus carriers; from these, a total of 34 S. aureus strains were isolated, of which 61.8% (n = 21) produced a strong biofilm. Phenotypic differences in strain biofilm production were detected in eight patients out of ten. All strains were sensitive to commonly used antibiotics. Broadly positive phage lytic activity (100%) was observed in six cocktails out of seven, and bacterial resistance towards phages was overcome using adaptation. Overall phages showed a promising in vitro effect in biofilm-forming S. aureus strains.

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BIOFILM FORMATION OF CANDIDA ALBICANS EXPOSED TO ETHANOL EXTRACT OF PROPOLIS

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2019.v11s1.18344 ◽

2019 ◽

pp. 210-213

Author(s):

ARIADNA ADISATTYA DJAIS ◽

JEMMY . ◽

NADHIFA PUTRI ◽

ANDIN RAHMANIA PUTRI ◽

RISQA RINA DARWITA ◽

...

Keyword(s):

Candida Albicans ◽

Biofilm Formation ◽

Fungal Infections ◽

Ethanol Extract ◽

Clinical Strain ◽

Propolis Extract ◽

Microtiter Plates ◽

Treatment And Prevention ◽

Student’S T

Objective: Propolis extract showed an excellent in vitro performance against yeast and was additionally found to be fungistatic and fungicidal. Propolisextract is also used for treatment and prevention of fungal infections. However, its effectiveness against Candida albicans biofilm formation requiresinvestigation. The study evaluated the ability of propolis to inhibit C. albicans while the fungus is growing as a biofilm in vitro.Methods: Two reference strains, C. albicans ATCC 25923 and a clinical strain (laboratory stock), were used in this study. For the biofilm experiment,the fungi were cultured in Tryptic Soy Broth medium with 1% sucrose and incubated at 37°C for 24 h, and different concentrations of ethanol extractof propolis were used as the inhibitor agents. Biofilm assays were performed in 96-well microtiter plates, quantification of the total biofilm biomasswas performed using a crystal violet staining method, and the Student’s t-test was chosen for statistical analyses.Results: Our data showed that 3 h incubation with propolis did not affect the biomass in the experimental group compared to the control. When theincubation time was extended to 18 h, the biomass increased significantly compared to the control.Conclusion: This study showed that several concentrations of propolis did not inhibit biofilm. However, in each incubation time, we observed nohyphal morphology in the biofilm mass. Propolis might attenuate the opportunistic virulence of fungus growing as a biofilm in vitro. Further studiesare necessary to confirm this phenomenon.

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