Minimum Fungicidal Concentration Research Articles

Antifungal activity of Morinda citrifolia methanolic extract against Candida albicans:: An in vitro study

Abstract Aim: Natural medicine used as an alternative and/or complementary treatment to counteract diseases is of great importance in public health. Therefore, the purpose of the present study was to assess the in vitro antifungal activity of Morinda citrifolia methanolic extract of peel, pulp, and seed against Candida albicans. Materials and Methods: The present study was experimental in vitro and cross-sectional. Eight replicates were prepared in Sabouraud dextrose agar with five wells each, where 0.12% chlorhexidine, distilled water, and methanolic extract of seed, peel, and pulp of Morinda citrifolia fruit were placed at concentrations of 10,690, 8,270, and 6,430 mg/mL, respectively, to evaluate sensitivity according to Duraffourd’s scale. In addition, the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were determined by dilution and agar seeding method. Statistical analysis was performed by analysis of variance (ANOVA) and Tukey’s post hoc test, considering a significance level of P < 0.05. Results: The inhibition halos of Morinda citrifolia methanolic extract of seed, peel, and pulp against Candida albicans measured on average 15.94, 11.94, and 11.56 mm, respectively. The MIC of seed, peel, and pulp extract were 1366.25, 2067.5, and 1607.5 mg/mL respectively, whereas the MFC for seed, peel, and pulp extract were 2672.50, 2067.5, and 3215 mg/mL, respectively. Moreover, seed extract presented significantly higher antifungal activity than peel and pulp (P < 0.001). Conclusions: Morinda citrifolia methanolic extract of peel, pulp, and seed showed fungistatic and fungicidal effect against Candida albicans, being this very sensitive to seed extract with a MIC of 1366.25 mg/mL and a MFC of 2672.5 mg/mL, which allows recommending the development of effective pharmacological formulations for the control of candidiasis.

Antifungal Effect and Durability of Chitosan Oligosaccharide Coating on Heat-cured Polymethylmethacrylate Surface

This study aimed to investigate the antifungal effect against Candida albicans (ATCC® 10231) and the durability of chitosan oligosaccharide (COS) coated on heat-polymerized polymethylmethacrylate (PMMA). The various concentrations of COS solution, 0.5, 1 and 2-fold of minimum fungicidal concentration (0.5, 1 and 2 MFC) were prepared and applied to PMMA specimens treated with sandblasting (SB), application of 3-aminopropyltriethoxysilane (APS), and a combination of both procedures (SB&APS). The toothbrush abrasion test was done to evaluate the remaining COS (n=10). The antifungal effect was determined through the colony count method (n=3). The number of fungal colonies was recorded at 16 and 48 hours. Data were analyzed using two-way and one-way ANOVA at the 95% confidence level. The result of the toothbrush abrasion test revealed that COS concentration and surface treatment affected coating durability. APS and SB&APS groups showed a high percentage of COS and significantly differed from SB groups (P = 0.00). For the antifungal effect, coatings with 1 MFC and 2 MFC demonstrated no difference, but coating with 0.5 MFC differed from others (P = 0.00). At 16 and 48 hours, PMMA coated with 1 MFC and 2 MFC by APS and SB&APS methods inhibited C.albicans. COS can be coated onto PMMA using APS and can inhibit C.albicans. With further development, this method could be applied to treat denture stomatitis alternatively to conventional antifungal drugs. Keywords: Candida albicans, Chitosan, Oligochitosan, Polymethylmethacrylate, Silanes

Antifungal activity of 2-chloro-N-phenylacetamide: a new molecule with fungicidal and antibiofilm activity against fluconazole-resistant Candida spp.

In the current context of emerging drug-resistant fungal pathogens such as Candida albicans and Candida parapsilosis, discovery of new antifungal agents is an urgent matter. This research aimed to evaluate the antifungal potential of 2-chloro-N-phenylacetamide against fluconazole-resistant clinical strains of C. albicans and C. parapsilosis. The antifungal activity of 2-chloro-N-phenylacetamide was evaluated in vitro by the determination of the minimum inhibitory concentration (MIC), minimum fungicidal concentration (MFC), inhibition of biofilm formation and its rupture, sorbitol and ergosterol assays, and association between this molecule and common antifungal drugs, amphotericin B and fluconazole. The test product inhibited all strains of C. albicans and C. parapsilosis, with a MIC ranging from 128 to 256 µg.mL-1, and a MFC of 512-1,024 µg.mL-1. It also inhibited up to 92% of biofilm formation and rupture of up to 87% of preformed biofilm. 2-chloro-N-phenylacetamide did not promote antifungal activity through binding to cellular membrane ergosterol nor it damages the fungal cell wall. Antagonism was observed when combining this substance with amphotericin B and fluconazole. The substance exhibited significant antifungal activity by inhibiting both planktonic cells and biofilm of fluconazole-resistant strains. Its combination with other antifungals should be avoided and its mechanism of action remains to be established.

Antifungal activity of Annona crassiflora Mart. dichloromethane fraction against strains of C. albicans.

Products derived from medicinal plants with antimicrobial activity are considered a promising alternative in the treatment of fungal infections. In this perspective, this study proposed to evaluate the antifungal activity of the dichloromethane fraction of Annona crassiflora Mart. against C. albicans strains. Tests were carried out to determine Minimum Inhibitory Concentration (MIC), Minimum Fungicide Concentration (MFC), microbial growth kinetics, fungal cell wall and membrane mechanisms of action, antifungal biofilm activity, and cytotoxic effects on human erythrocytes. The extract presented MIC and MFC values that ranged from 256 µg/mL to 1,024 µg/mL, with fungicidal activity in the microbial growth kinetics assay. The mechanism of action did not occur through damage to the cell wall or via binding to ergosterol in the membrane, though the fraction presents activity against biofilm and is not cytotoxic in human erythrocytes. The dichloromethane fraction of Annona crassiflora Mart. presented antifungal activity and reduced biofilm growth, without toxicity against human erythrocytes; however, further studies are needed to define its mechanism of action.

In vitro antifungal activity of MMV Pathogen Box® compounds alone or in combination with antifungal drugs against mucormycosis agents

Mucormycosis is a severe fungal infection that demands immediate and decisive intervention upon suspicion. The causative agents of mucormycosis exhibit inherent resistance to echinocandins and voriconazole, and their in vitro susceptibility to terbinafine is highly variable and species-specific. Considering these factors and the limitations of currently available antifungal therapies, the identification of novel antifungals with potent activity against mucormycosis is of paramount importance. This study aims to identify compounds from the MMV Pathogen Box® presenting antifungal activity against selected mucormycosis agents and to evaluate their potential synergistic effects when combined with antifungal drugs. A screening of the Pathogen Box® compounds was conducted, isolated or in combination with sub-inhibitory concentrations of amphotericin B, isavuconazole or posaconazole, against a Rhizopus oryzae strain. Hits from the screenings were further evaluated against eight Mucoralean strains for minimal inhibitory and fungicidal concentration determinations and to confirm synergistic interactions using the checkerboard method. Ultrastructural studies were performed using scanning electron microscopy. MMV675968 exhibited fungicidal activity against a R. oryzae strain. All but one Rhizopus spp. strains presented MIC ≤ 1 μg/mL, with a geometric mean of 0.78 μg/mL observed across all isolates for this compound, which did not change significantly the cellular structure of this fungus. The combination screening with antifungal drugs revealed six additional compounds potentially active against the R. oryzae strain, two of them demonstrated proven synergism through the checkerboard assay. This first study with the MMV Pathogen Box® and Zigomycetes highlights promising new treatment options for mucormycosis in the future.

Production of new ent-hardwickiic acid derivatives by microbial transformation and their antifungal activity

Ent-hardwickiic acid is the major compound of Copaifera pubiflora Benth oleoresin traditionally used in Brazilian folk medicine as an antimicrobial agent. Microbial transformation of ent-hardwickiic by Cunninghamella elegans ATCC 10028b resulted in two and five antifungal derivatives (four new ones) produced in the Czapek modified and Koch's K1 media, respectively. The derivatives were isolated and their structures were determined by spectral analysis, namely 1D/2D NMR and HR-ESIMS. All compounds were tested for cytotoxic and antifungal activities and they were not cytotoxic to the tested cell lines, but all derivatives showed fungicidal activity against Candida glabrata and Candida krusei, which have emerged as resistant to fluconazole. One of the yet unreported biotransformation products displayed the strongest activity with minimum fungicidal concentration values smaller than the other compounds, including fluconazole.

ANTIMICROBIAL AND ANTIBIOFILM PROPERTIES OF Combretum platypetalum subsp. oatesii (Rolfe) Exell (Combretaceae) EXTRACTS

Combretum species exhibit potent antibacterial and antifungal properties, but limited studies exist for most species. This study reports for the first time the in vitro antimicrobial and antibiofilm activities of Combretum platypetalum leaf, stem, and root. Fungi and bacterial strains implicated in the causes of some of the conditions to which C. platypetalum is traditionally applied were evaluated by Agar-well diffusion and micro-dilution methods, and their biofilm inhibition capacities were investigated. The highest zone of fungal inhibition of 22.70 ± 2.08 mm against C. albicans and the highest zone of bacterial inhibition of 21.70 ± 2.08 mm against S. aureus were observed. The minimum inhibitory concentration (MIC) ranged between 0.0156 mg/mL and 8.0000 mg/mL for fungi and 0.3906 mg/mL and 8.0000 mg/mL for bacteria at P = 0.0002. The best minimum fungicidal concentrations (MFCs), at P = 0.003, were observed at 0.0630 mg/mL for the leaf against C. glabrata and C. neoformans and 0.5 mg/mL for the root against C. neoformans, respectively. The highest biofilm inhibition of 40.61%, 36.04%, and 23.85% was observed in the stem, root, and leaf extracts against C. glabrata. Chloroform leaf extract reveals ?-sitosterol as the active antifungal and antibacterial agent. Computational antifungal activity and molecular docking analysis of the structure versus target (sterol 14?- demethylase) show a binding affinity for druggability. The leaves and roots are the most potent parts of the plant, with leaf extracts showing non-synergistic effects. Bioguided isolation is recommended to fully explore the species' evidence-based therapeutic potential.

ROS mediated anticandidal efficacy of 3-Bromopyruvate prevents vulvovaginal candidiasis in mice model.

Candidal infections, particularly vulvovaginal candidiasis (VVC), necessitate effective therapeutic interventions in clinical settings owing to their intricate clinical nature and elusive understanding of their etiological mechanisms. Given the challenges in developing effective antifungal therapies, the strategy of repurposing existing pharmaceuticals has emerged as a promising approach to combat drug-resistant fungi. In this regard, the current study investigates molecular insights on the anti-candidal efficacy of a well-proven anticancer small molecule -3-bromopyruvate (3BP) against three clinically significant VVC causing Candida species viz., C. albicans, C. tropicalis and C. glabrata. Furthermore, the study validates 3BP's therapeutic application by developing it as a vaginal cream for the treatment of VVC. 3BP exhibited phenomenal antifungal efficacy (killing >99%) with minimum inhibitory concentrations (MIC) and minimum fungicidal concentrations (MFC) of 256 μg/mL against all tested Candida spp. Time killing kinetics experiment revealed 20 min as the minimum time required for 3BP at 2XMIC to achieve complete-killing (99.9%) in all Candida strains. Moreover, the ergosterol or sorbitol experiment explicated that the antifungal activity of 3BP does not stem from targeting the cell wall or the membrane component ergosterol. Instead, 3BP was observed to instigate a sequence of pre-apoptotic cascade events, such as phosphatidylserine (PS) externalization, nuclear condensation and ROS accumulations, as evidenced by PI, DAPI and DCFH-DA staining methods. Furthermore, 3BP demonstrated a remarkable efficacy in eradicating mature biofilms of Candida spp., achieving a maximum eradication level of 90%. Toxicity/safety profiling in both in vitro erythrocyte lysis and in vivo Galleria mellonella survival assay authenticated the non-toxic nature of 3BP up to 512 μg/mL. Finally, a vaginal cream formulated with 3BP was found to be effective in VVC-induced female mice model, as it significantly decreasing fungal load and protecting vaginal mucosa. Concomitantly, the present study serves as a clear demonstration of antifungal mechanistic action of anticancer drug -3BP, against Candida species. This finding holds significant potential for mitigating candidal infections, particularly VVC, within healthcare environments.

Study on the inhibitory effect of fermentation extract of Microporus vernicipes on Candida albicans.

Candida albicans is one of the most common species of Candida, which cause various mucosal and systemic infectious diseases. However, the resistance rate to existing clinical antifungal drugs gradually increases in C. albicans. Therefore, new antifungal drugs must be developed to solve the current problem. This study discovered that the solid fermented ethyl acetate crude extract of Microporus vernicipes had inhibitory activity on C. albicans. This study determined that the Mv5 components had significantly inhibited the activity of C. albicansusing column chromatography separation component screening. The components included 23 compounds of fatty acids and their derivatives, alkaloids, phenols, and other classes using ultra-high performance liquid chromatography tandem high-resolution mass spectrometry (UHPLC-HR-MS) analysis, with fatty acids constituting the primary components. The mechanism of action showed that the minimum inhibitory concentration (MIC) of Mv5 components against C. albicans was 15.63μg/mL, while minimum fungicidal concentration (MFC) was 31.25μg/mL. Mv5 components can inhibit the early biofilm formation and destroy the mature biofilm structure. It can inhibit the germ tube growth of C. albicans, thereby inhibiting the transformation of yeast morphology to hyphae. We detected 193 differentially expressed genes, including 156 upregulated and 37 downregulated genes in the Mv5 components of the MIC concentration group. We detected 391 differentially expressed genes, including 334 upregulated and 57 downregulated expression genes in the MFC concentration group. Among these differentially expressed genes, the genes related to mycelium and biofilm formation were significantly downregulated. GO enrichment analysis presented that single-organism process metabolic process, and cellular processes were the biological processes with the most gene enrichment. Kyoto Encyclopedia of Genes and Genomes (KEGG)of Mv5 components were mainly enriched in metabolic pathways, such as meiosis yeast and amino acid metabolism. Therefore, it is believed that the fermentation extract of M. vernicipes inhibits C. albicans, which can provide clues for developing effective antifungal drugs.

Anti-fungal Activity of 4 Herbal Mouthwashes vs. Chlorhexidine Against Candida albicans and C. glabrata: A Preliminary Study

Anti-fungal Activity of 4 Herbal Mouthwashes vs. Chlorhexidine Against Candida albicans and C. glabrata: A Preliminary Study

Antifungal Properties of Pimenta racemosa (Mill.) and Mentha x piperita (L.) Essential Oils against Fusarium oxysporum Causing Tomato Fruit Rot

Fungal pathogens are responsible for the rot of tomatoes and cause health hazards to consumers as well as economic loss. The aims of this study were to evaluate the efficacy of essential oils from Pimenta racemosa (Mill.) and Mentha x piperita (L.) leaves as plant-based fungicide against Fusarium oxysporum strains associated with tomato rot. The chemical composition of the essential oils was determined by GC-MS analysis. Antifungal effects of essential oils against strain of F. oxysporum obtained from rotted tomato, were evaluated through the determination of Minimal Fungicide Concentration (MFC) and the Minimal Inhibitory Concentration (MIC). Results indicated that β-myrcene (19.48%), chavicol (15.39 %), and eugenol (46.65%) were the major components of the essential oil of Pimenta racemosa (Mill.). L-menthone (17.19 %), menthol (24.79%), and menthofuran (26.78 %) were the major components of the essential oil of Mentha x piperita (L.). Essential oil of Pimenta racemosa (Mill.) exhibited the highest antifungal activity against the growth of Fusarium oxysporum strain tested with a MIC and MFC of 0.75 and 1.25 μl.ml-1 respectively. These results provide an experimental basis for further research on the use of these plant extracts in the development of eco-friendly preservatives against fungi causing rots of tomato in post-harvest.

Antifungal Activity of Chitosan Polymeric Nanoparticles and Correlation with Their pH Against Mucor circinelloides Causing Mucormycosis, Along with Penicillium notatum and Aspergillus Species

Mucormycosis is uncommon, yet it is more prevalent among individuals with underlying health conditions and those who are immunocompromised. Chitosan is studied because of its appealing properties and diverse applications. The purpose of this work is to synthesize chitosan nanoparticles (CSNPs) by ionic gelation method at various pH levels and test them against Mucor and other filamentous fungus. Field Emission Scanning Electron Microscope, Zeta sizer, Zeta potential, and Fourier Transformed Infrared Spectroscopy were used to characterize CSNPs. Hydrodynamic size increased considerably with increasing pH. Our CSNPs were tested against fungal isolates of Aspergillus Flavus RCMB 02783, Aspergillus Fumigatus RCMB 02564, and Aspergillus Niger RCMB 02588, Penicillium Notatum (NCPF 2881) and (M. circinelloides CNRMA 03.894) causing mucromycosis. Antifungal activity was investigated using Minimum inhibitory concentration (MIC), Minimum Fungicidal concentration (MFC), Disc diffusion assay, and Antifungal inhibitory percentages methods. The best antifungal efficacy results were obtained through CSNPs prepared at pH = 4.4 at very low concentration for MIC (1.03 or 2.75 μg/mL) with 100% M. circinelloides inhibition followed by pH = 4.6 with MIC (73 or 208 μg/mL) and 93% M. cirecinelloides inhibition %. Future usage of these materials in masks or wound dressing to avoid fungal infections, including mucormycosis following COVID-19, penicillium, and aspergillosis toxicity and infections.Graphical

Antifungal activity of cercosporamide produced by Phaeosphaeriaceae GV-1 against pathogenic fungi.

Fungal infections affect millions of people worldwide, and the several cases are related to invasive infections, which is a problem mainly for immunocompromised people, such as transplant and cancer patients with high mortality and morbidity rates. In addition, the number of emerging and multidrug-resistant fungal species has increased in the last decade. The search for new antifungal compounds is necessary, due to the increase in cases of resistance and the toxicity of drugs used in fungal infection treatment. This work aimed to study the antifungal activity of cercosporamide produced by Phaeosphaeriaceae GV-1. Cercosporamide was tested against pathogenic fungi by determining the minimum inhibitory (MIC) and minimum fungicidal (MFC) concentrations, using the broth microdilution method. Cercosporamide showed antifungal activity in vitro against 13 of 16 strains of medical importance tested, with the most susceptible species being Candida tropicalis, with MIC and MFC of 15.6 μg/mL. Thus, cercosporamide might be considered a promising therapeutic antifungal agent.

PHYTOCHEMICAL COMPONENTS, PROXIMATE ANALYSIS, AND ANTIMICROBIAL PROPERTIES OF FERMENTED AND NON-FERMENTED SAMPLES OF Zingiber officinale

The desirability of fermented foods may be attributed to their benefits. This study investigated the pH, in vitro antimicrobial assays including Minimum Inhibition Concentration (MIC), Minimum Fungicidal Concentration (MFC), and disc diffusion assay of fermented and non-fermented samples of Zingiber officinale against Candida albicans, Staphylococcus aureus, and Escherichia coli. But phytochemical screening, proximate compositions, and High-Performance Liquid Chromatography (HPLC) were carried out only on the non-fermented sample. The pH decreased steadily throughout while the MIC ranged between 12.5 – 50.0 mg/mL. The MIC of the non-fermented sample ranged between 25-50.0 mg/Ml. The highest zones of inhibition obtained were between 8.0 mm- 16.0 mm at 100 mg/mL, 8.0 mm- 14.00 mm at 50.0 mg/mL, and 2.00-8.4 mm at 25 mg/mL. Comparatively, the zones were higher (6.8-8.4 mm) for the fermented than for the non-fermented sample (1.8-4.0 mm) and higher against S. aureus (4-16.0 mm) than E. coli (2.0 -12.0 mm) and C. albicans (2.0-6.0 mm). The fermented sample contained alkaloids, anthraquinones, flavonoids, phlobatannins, saponins, tannins, and terpenoids, while alkaloids, flavonoids, glycosides, saponins, steroids, tannins, and terpenoids were in the non-fermented sample. The fermented sample contained 87.31% moisture, 1.24% ash, 0.29% fat, 9.73% carbohydrate, 0.24% fiber, and 1.48% protein, while the non-fermented sample contained 83.10% moisture, 1.02% ash, 0.21% fat, 14.46% carbohydrate, 0.18% fiber, and 1.24% protein. HPLC revealed Alpha farnesene (51.77), zingiberene (11.30), zingerone (14.53), 6-gingerol (12.44), and quercetin (35.21) in the fermented sample. Thus, the fermented sample of Zingiber officinale is a suitable candidate for the promotion of human health.

Antifungal susceptibility testing following the CLSI M27 document, along with the measurement of MFC/MIC ratio, could be the optimal approach to detect amphotericin B resistance in Clavispora (Candida) lusitaniae. Susceptibility patterns of contemporary isolates of this species.

Antifungal susceptibility testing (AST) is crucial in clinical settings to guide appropriate therapy. Nevertheless, discrepancies between treatment response and some results still persist, particularly in detecting resistance to amphotericin B (AMB) in Clavispora (Candida) lusitaniae. This study aimed to assess the susceptibility patterns of 48 recent isolates of C. lusitaniae to 9 antifungal agents and explore the feasibility of using a CLSI reference-based method to identify AMB resistance. Microdilution techniques revealed a wide range of minimal inhibitory concentration (MIC) values for azole antifungals, while echinocandins and AMB exhibited a narrow range of MIC values, with all strains considered wild-type for the tested polyene and echinocandins. However, when agar diffusion (ellipsometry) was employed for AST, certain strains displayed colonies within the inhibition ellipse, indicating potential resistance. Interestingly, these strains did not respond to AMB treatment and were isolated during AMB treatment (breakthrough). Moreover, the evaluation of AMB minimum fungicidal concentrations (MFCs) indicated that only the strains with colonies inside the ellipse had MFC/MIC ratios ≥ 4, suggesting reduced fungicidal activity. In conclusion, this study confirms the effectiveness of ellipsometry with RPMI-1640 2% glucose agar for detecting AMB resistance in C. lusitaniae. Additionally, the proposed approach of culturing "clear" wells in the microdilution method can aid in uncovering resistant strains. The findings highlight the importance of appropriate AST methods to guide effective treatment strategies for deep-seated candidiasis caused by C. lusitaniae. Further collaborative studies are warranted to validate these findings and improve the detection of AMB clinical resistance.

Antifungal Potential of Propolis Extract Against Candida Albicans: Literature Review

The increasing rate of Candida resistance against pharmacological drugs has become a particular focus in the medical world, especially considering the high mortality rate due to systemic infections caused by Candida. It is evident that the urgency of developing alternative medicines applies for potential natural ingredients like propolis. This literature review aims to discuss the antifungal effectiveness of propolis against C. albicans which is described by the MIC (minimum inhibitory concentration) and MFC (minimum fungicidal concentration) values from the test results. The search for relevant literature was conducted using the Google Scholar, ScienceDirect, and PubMed databases using the keywords "Propolis AND MIC AND MFC AND Candida albicans". There were nine articles that had been adjusted to the inclusion and exclusion criteria based on the PICOS Framework. The propolis in the article uses various solvents with various extraction methods, and various antifungal tests are carried out on standardized and clinical isolates of C. albicans. All the reviewed articles consistently support the finding that propolis has a strong ability to inhibit the growth and kill the pathogenic fungus C. albicans. The lowest MIC value found was 0.06 μg/mL, and the MFC value was 0.121 μg/mL. Propolis has potential as an ingredient that can be further developed as a non-pharmacological therapy for pathogenic fungal infections.

Fungicidal and antibiofilm activities of 2-Bromo-N-Phenylacetamide Against Fluconazole-Resistant Candida spp.: a promising antifungal agent

Purpose: Amides are proven to possess antimicrobial properties against both fungi and bacteria. The synthetic amide 2-bromo-N-phenylacetamide exhibits antifungal activity according to scientific literature, however, data remains scarce. Given the clinical and epidemiological importance as well as the increasing antifungal resistance in Candida spp., here we aimed to better explore the antifungal potential of 2-bromo-N-phenylacetamide against fluconazole-resistant Candida albicans, Candida glabrata, Candida parapsilosis and for the first time, against Candida tropicalis as well. Methods: Minimum Inhibitory and Minimum Fungicidal Concentrations (MIC/MFC) were established by a Broth Microdilution Technique. Mode of action was explored by Ergosterol and Sorbitol assays. For investigating the combined effect between 2-bromo-N-phenylacetamide and the standard antifungals Amphotericin B and Fluconazole a Checkerboard Technique was performed. Finally, the antibiofilm properties of 2-bromo-N-phenylacetamide were assessed by a Crystal Violet Staining Assay. Results: 2-bromo-N-phenylacetamide exhibited a fungicidal effect, MIC of 32 µg.mL-1 (87.5% of the strains) and MFC of 64 µg.mL-1 (81.25% of the strains). It also demonstrated an activity against Candida albicans mature biofilms as effective as that for Amphotericin B at all tested concentrations. When associated with Fluconazole and Amphotericin B it exerted indifferent effects. Mode of action remained unclear. Conclusion: The synthetic amide 2-bromo-N-phenylacetamide possesses a strong antifungal activity against Candida albicans, Candida tropicalis, Candida glabrata and Candida parapsilosis and an antibiofilm property against Candida albicans as effective as that exhibited for Amphotericin B.

In Vitro Evaluation of Corn Silk Extract as a Potential Alternative Therapy for Vulvovaginal Candidiasis

Abstract Aim To determine the effectivity of corn silk extract in inhibiting the growth of C. albicans. Materials and Methods Corn silk used in this study was Zea mays L. var. indurata and was collected from Desa Suka Makmur, Deli Serdang, Sumatera Utara Province, Indonesia. Tested C. albicans is C. albicans from VVC patients, obtained from the Microbiology Laboratory of Faculty of Medicine, Universitas Sumatera Utara. Preparation of this study included corn silk extraction by maceration method with 96% ethanol, followed by dilution with 10% DMSO (Dimethyl Sulfoxide) into concentrations of 75%, 50%, 25%, and 12.5%. Well diffusion method was used to detect growth inhibition of C. albicans and agar dilution was used to determine MIC (Minimum Inhibitory Concentration) and MFC (Minimum Fungicidal Concentration) of corn silk extract. Data were statistically analyzed with Kruskal-Wallis analysis. Results From this study, corn silk extract of all concentrations showed an inhibitory effect against C. albicans. The mean diameter of inhibition zones for each concentration respectively from highest to lowest concentration were 27.4875 ± 0.3838 mm, 26.7250 ± 0.2533 mm, 25.7250 ± 0.2598 mm, and 24.9375 ± 0.2462 mm. Kruskal-Wallis analysis showed significant results (p-value 0.001). MIC and MFC were 1.5625% and 6.25%, respectively. Conclusions Corn silk extract of concentrations 75%, 50%, 25%, and 12,5% have antifungal activity against C. albicans. The higher the concentration of corn silk extract, the higher the efficacy in inhibiting the growth of C. albicans.

A combinatorial approach of Monarda citriodora essential oil (MEO) and linalool vapors to control fruit rot of Citrus limon caused by a new pathogen, Aspergillus foetidus, and its underlying mode of action.

Citrus limon (lemon) is a widely cultivated citrus fruit. Significant postharvest losses due to fungi plague its production. Environmental and human health hazards have made the application of synthetic fungicides unsuitable. Despite the previous reports of antifungal activities of essential oil (EO) vapors, their synergistic combinations are understudied. Synergistic vapor combinations are advantageous due to less concentration of active components. This study aimed to isolate and identify postharvest fungal pathogens lemon and to evaluate the antifungal effects of synergistic Monarda citriodora EO (MEO)-constituent vapor combinations in vivo and in vitro. Postharvest fungal pathogens of lemon (C. limon) were isolated from various infected samples. The most pathogenic isolate was identified through morphology and its ITS-based rRNA gene sequencing as Aspergillus foetidus (O4). This is the first report of A. foetidus as a postharvest pathogen of lemon. The minimum fungicidal concentrations (MFCs) of MEO vapors treatment against O4 were 1346.15µL/L air. For carvacrol, hexanal, and linalool, MFC was same (96.16µL/L air). Checkerboard assays demonstrated that 1/4 MFC of MEO (336.54µL/L air) and 1/4 MFC of linalool (24.04µL/L air) (M+L) were synergistic against O4. M+L vapors reduced the O4 growth on lemons during storage by 64%±1.50% and preserved their quality (low weight loss %, unchanged pH, increased ascorbic acid content). Propidium iodide staining, ergosterol content analysis, calcofluor white staining and chitin content analysis revealed the integrity loss of the O4 plasma membrane and cell wall. 2',7'-Dichlorofluorescin diacetate staining revealed accumulation of intracellular reactive oxygen species (ROS), and scanning electron microscopy (SEM) analysis exposed the M+L treated mycelia with malformations. M+L vapors offer protection for lemons from A. foetidus and preserve their quality during storage.

Berberine-fluconazole microparticle-based combination therapy to treat candidiasis infections.

This study aims to incorporate alginate microparticles containing berberine and fluconazole into two different types of pharmaceutical formulations, to subsequently evaluate the antifungal activity against Candida albicans. Alginate microparticles containing BBR (berberine) and FLU (fluconazole) were produced by the spray-drying technique, characterized and incorporated in two pharmaceutical formulations, a vaginal cream and artificial saliva. Broth microdilution, checkerboard, time-kill curve, and scanning electron microscopy were carried out to determine the antifungal effects of BBR and FLU against C. albicans. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of free BBR were 125μg ml-1. Synergism between BBR and FLU was demonstrated by a fractional inhibitory concentration index (FICI)=0.0762. The time-kill curve for the combination BBR+FLU showed a more pronounced decrease in fungal growth in comparison to free drugs, and an antibiofilm effect of BBR occurred in the formation and preformed biofilm. Alginate microparticles containing BBR and FLU were obtained and incorporated in a vaginal cream and artificial saliva. Both formulations showed good stability, antifungal effects, and organoleptic characteristics, which suggest that BBR-FLU microparticles in formulations have potential as antifungal therapy.