Antifungal Drugs Amphotericin Research Articles

Anfotericina B liposomal: farmacología clínica, farmacocinética y farmacodinamia

Liposomal amphotericin B is a lipid formulation of the antifungal drug amphotericin B with some distinguishing characteristics in its pharmacological behavior that entail some clinical differences of great interest. The significant improvement in the systemic and renal tolerability is one of them. This fact is related to the great stability of the liposome, promoted by its negative charge, the presence of cholesterol and the remarkable thermo-stability of the remaining lipids that compose it. In this situation, amphotericin B seems to be released from the liposome not spontaneously but when the liposome binds to the ergosterol in the fungal cell membrane. For this reason, there is almost no free amphotericin B in plasma or tissues, although it seems that its availability is greater when there is fungal infection. As a consequence, when the pharmacokinetic behavior is studied, the concentration and availability of liposomal amphotericin B are very high, and its volume of distribution is reduced in comparison with the other formulations.

“Metal-free” synthesis and antifungal activity of 3-thiocyanatopyrazolo[1,5-a]pyrimidines

A convenient stepwise synthesis of substituted 3-thiocyanatopyrazolo[1,5-a]pyrimidines under mild conditions using available reagents and solvents was proposed. A number of these compounds were found to possess pronounced and selective antifungal activity, which is comparable or superior to the activity of the antifungal drugs amphotericin B and fluconazole.

The Antifungal Mechanism of Amphotericin B Elucidated in Ergosterol and Cholesterol-Containing Membranes Using Neutron Reflectometry.

We have characterized and compared the structures of ergosterol- and cholesterol-containing 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membranes before and after interaction with the amphiphilic antifungal drug amphotericin B (AmB) using neutron reflection. AmB inserts into both pure POPC and sterol-containing membranes in the lipid chain region and does not significantly perturb the structure of pure POPC membranes. By selective per-deuteration of the lipids/sterols, we show that AmB extracts ergosterol but not cholesterol from the bilayers and inserts to a much higher degree in the cholesterol-containing membranes. Ergosterol extraction by AmB is accompanied by membrane thinning. Our results provide new insights into the mechanism and antifungal effect of AmB in these simple models of fungal and mammalian membranes and help understand the molecular origin of its selectivity and toxic side effects.

Candida metrosideri pro tempore sp. nov. and Candida ohialehuae pro tempore sp. nov., two antifungal-resistant yeasts associated with Metrosideros polymorpha flowers in Hawaii.

Flowers produce an array of nutrient-rich exudates in which microbes can thrive, making them hotspots for microbial abundance and diversity. During a diversity study of yeasts inhabiting the flowers of Metrosideros polymorpha (Myrtaceae) in the Hawai’i Volcanoes National Park (HI, USA), five isolates were found to represent two novel species. Morphological and physiological characterization, and sequence analysis of the small subunit ribosomal RNA (rRNA) genes, the D1/D2 domains of the large subunit rRNA genes, the internal transcribed spacer (ITS) regions, and the genes encoding the largest and second largest subunits of the RNA polymerase II (RPB1 and RPB2, respectively), classified both species in the family Metschnikowiaceae, and we propose the names Candida metrosideri pro tempore sp. nov. (JK22T = CBS 16091 = MUCL 57821) and Candida ohialehuae pro tempore sp. nov. (JK58.2T = CBS 16092 = MUCL 57822) for such new taxa. Both novel Candida species form a well-supported subclade in the Metschnikowiaceae containing species associated with insects, flowers, and a few species of clinical importance. The ascosporic state of the novel species was not observed. The two novel yeast species showed elevated minimum inhibitory concentrations to the antifungal drug amphotericin B (>4 μg/mL). The ecology and phylogenetic relationships of C. metrosideri and C. ohialehuae are also discussed.

Carvacrol essential oil stimulates growth performance, immune response, and tolerance of Nile tilapia to Cryptococcus uniguttulatus infection.

This study investigated the antifungal activity of 5 essential oils (EOs) towards yeasts recovered from diseased fishes; and focused on the efficacy of one EO (carvacrol) on growth performance, non-specific immunity, and disease resistance of Nile tilapia Oreochromis niloticus against Cryptococcus uniguttulatus challenge. Thymoquinone, thymol, carvacrol, eugenol, and cinnamon were first tested in vitro against 20 clinical yeast strains in comparison with antifungal drugs (fluconazole, ketoconazole, itraconazole, amphotericin B, nystatin, and clotrimazole) using disc diffusion and broth microdilution methods. For the in vivo challenge, fish (n = 150) were divided into 5 groups (carvacrol prophylaxis, carvacrol treatment, itraconazole treatment, unchallenged control, and positive control; 30 fish group-1) with 3 replicates. Phagocytic activity, reactive oxygen species production, reactive nitrogen species production, myeloperoxidase, lysozyme activity, and total immunoglobulins were tested before and after challenge. Relative percent survival (RPS) and mortality percent were determined as indicators for functional immunity. EOs displayed divergent degrees of antifungal activity, and carvacrol was the most effective against the tested yeasts. The dietary additive of carvacrol significantly enhanced growth performance, all immunological parameters, and the RPS values (90%) compared to other treatments. This unique experimental model indicates that carvacrol seems promising not only for enhancing immunity and promoting fish growth, but also for controlling emerging fungal infections. Future studies should investigate different concentrations of carvacrol as well as its antifungal activity in different fish species.

Desorption Properties and Bactericidal and Fungicidal Activity of Nanostructured Coatings Based on Hexagonal Boron Nitride Saturated with Therapeutic Preparations

The study is aimed at creating nanostructured coatings based on hexagonal boron nitride with a high sorption capacity with respect to various therapeutic drugs and prolonged antibacterial effect due to gradual release into the physiological environment. The coatings were obtained via interaction of gaseous ammonia with a precursor of amorphous boron nanoparticles. The antibiotics gentamicin sulfate and ciprofloxacin and the antifungal drug amphotericin B were used as bactericides. The release kinetics of the antibiotics were assayed for different pH values. Antibacterial studies showed that all types of coatings are bactericidal to the antibiotic-resistant strain Escherichia coli K-261. Coatings saturated with amphotericin B showed antifungal activity against various ascomycete strains Neurospora crassa wild type wt-987 and nitrogen metabolism mutants nit-2 and nit-6.

Engineering Nanogels for Drug Delivery to Pathogenic Fungi Aspergillus fumigatus by Tuning Polymer Amphiphilicity.

Invasive aspergillosis is a serious threat to immunodeficient and critically ill patients caused mainly by the fungus Aspergillus fumigatus. Here, poly(glycidol)-based nanogels (NGs) are proposed as delivery vehicles for antifungal agents for sustained drug release. NGs are formed by simple self-assembly of random copolymers, followed by oxidative cross-linking of thiol functionalities. We investigate the impact of copolymer amphiphilicity on NG interaction with mature fungal hyphae in order to select the optimal drug delivery system for model antifungal drug amphotericin B. The results show that drug-loaded NGs decrease minimal inhibitory concentration (MIC) for around four times and slow down the fungal biofilm synthesis at concentrations lower than MIC. Our results suggest that amphiphilicity of nanoparticle's polymer matrix is an important factor in understanding the action of nanocarriers toward fungal cells and should be considered in the development of nanoparticle-based antifungal therapy.

Selective antifungal activity of silver nanoparticles: A comparative study between Candida tropicalis and Saccharomyces boulardii

In this study, the antifungal activity of AgNPs was tested against C. tropicalis (pathogen fungi) and S. boulardii (probiotic). The effectiveness of the AgNPs was assessed by comparing their antifungal activity with a triazole antifungal drug fluconazole and amphotericin B. The AgNPs have a polygonal-like shape (average size of 35 ± 15 nm) with 1.2% wt. of metallic silver stabilized with 18.8% wt. of polyvinylpyrrolidone (PVP) in 80% wt. of distilled water. The results revealed that 35 μg/mL of fluconazole inhibits 55–60% of both fungal cell growth. As for amphotericin B, 5 μg/mL is sufficient to inhibit more than 95% of both fungal cells. For AgNPs, 25 μg/mL was needed to inhibit 90% of the C. tropicalis cell growth, but remarkably, 50% of the S. boulardii cell population remains viable, which can potentiate cell reproduction. Our results could initiate the development of AgNPs possessing selective specificity against pathogenic fungal species.

Multi-locus Sequence Typing and Whole Genome Sequence Analysis of Cryptococcus neoformans Isolated from Clinical Specimens in Vajira Hospital, Bangkok, Thailand.

The basidiomycete yeast Cryptococcus neoformans causes disease in immunocompromized patients. Whole genome sequencing (WGS) technology provides insights into the molecular epidemiology of C. neoformans. However, the number of such studies is limited. Here we used WGS and multilocus sequence typing (MLST) to determine the genetic diversity of C. neoformans isolates and genetic structures of their populations among patients admitted to a single hospital in Bangkok, Thailand. Seven isolates from six patients collected during 1year were identified as C. neoformans sensu stricto according to colony morphology, microscopy, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nucleotide sequence analysis of internal transcribed sequences. These isolates were sensitive to the antifungal drugs amphotericin B, fluconazole, 5-flucytosine, voriconazole, itraconazole and posaconazole and were mating type α and molecular type VNI. MLST analysis identified ST4, ST5 and ST6. We further employed WGS to determine the genetic diversity and relationships of C. neoformans isolated here combined with C. neoformans sequences data acquired from a public database (n = 42). We used the data to construct a phylogenetic tree. WGS provided additional genomics data and achieved high discriminatory power for identifying C. neoformans isolates isolated in Thailand. This report further demonstrates the applicability of WGS analysis for conducting molecular epidemiology and provides insight into the genetic diversity of C. neoformans isolates from one hospital in Thailand.

Inhibitory Effects of Antifungal Drugs on Biofilm Producing Aspergillus spp. Recovered from Drinking Water System

Aims: Biofilms formed in drinking water distribution systems serve as a continuous source of fungal infections. Biofilms are thick aggregates of adherent microorganisms including pathogenic species of fungi. Respiratory diseases and skin allergy reactions are caused by drinking water containing biofilm forming fungus and bacteria. One of the main causes of nosocomial infections and respiratory diseases in hospitals is due to the fungal biofilm formation in machines, catheters and other surgical instruments. There are some antifungal drugs which are used to control biofilm formation to minimize the infection rate. Methodology and results: The present study was conducted to isolate and identify Aspergillus species which are the main fungal spp. responsible for the biofilm formation in drinking water and to check their antifungal susceptibility against antifungal drugs. The isolated fungal samples from drinking water were cultivated on Potato dextrose agar for the isolation of Aspergillus species. Isolated Aspergillus species were identified on the basis of cultural, morphological and microscopic examination. Then in-vitro ability of biofilm produced by isolated Aspergillus species was estimated using microtitre plate method and quantification by crystal violet assay. Antifungal susceptibility testing against isolated fungal spp. was done by antifungal drug Amphotericin B. Results: From results, it is concluded that drinking water of labs, hospitals and common water chillers were more prevelant by Aspergillus species whereas water from reverse osmosis plants showed negative results. From microtitre plate method and crystal violet assay, it was concluded that Aspergillus spp. are Susceptible against Amphotericin B drug as compared to miconazole. Keywords: Aspergillus spp; Biofilm; Drinking water; Disk diffusion method; Amphotericin B

Antiamoebic activity of synthetic tetrazoles against Acanthamoeba castellanii belonging to T4 genotype and effects of conjugation with silver nanoparticles.

Acanthamoeba causes diseases such as Acanthamoeba keratitis (AK) which leads to permanent blindness and granulomatous Acanthamoeba encephalitis (GAE) where there is formation of granulomas in the brain. Current treatments such as chlorhexidine, diamidines, and azoles either exhibit undesirable side effects or require immediate and prolonged treatment for the drug to be effective or prevent relapse. Previously, antifungal drugs amphotericin B, nystatin, and fluconazole-conjugated silver with nanoparticles have shown significantly increased activity against Acanthamoeba castellanii. In this study, two functionally diverse tetrazoles were synthesized, namely 5-(3-4-dimethoxyphenyl)-1H-tetrazole and 1-(3-methoxyphenyl)-5-phenoxy-1H-tetrazole, denoted by T1 and T2 respectively. These compounds were evaluated for anti-Acanthamoeba effects at different concentrations ranging from 5 to 50μM. Furthermore, these compounds were conjugated with silver nanoparticles (AgNPs) to enhance their efficacy. Particle size analysis showed that T1-AgNPs and T2-AgNPs had an average size of 52 and 70nm respectively. After the successful synthesis and characterization of tetrazoles and tetrazole-conjugated AgNPs, they were subjected to anti-Acanthamoeba studies. Amoebicidal assay showed that at concentration 10μM and above, T2 showed promising antiamoebic activities between the two compounds while encystation and excystation assays reveal that both T1 and T2 have inhibited differentiation activity against Acanthamoeba castellanii. Conjugation of T1 and T2 to AgNP also increased efficacy of tetrazoles as anti-Acanthamoeba agents. This may be due to the increased bioavailability as AgNP allows better delivery of treatment compounds to A. castellanii. Human cell cytotoxicity assay revealed that tetrazoles and AgNPs are significantly less toxic towards human cells compared with chlorhexidine which is known to cause undesirable side effects. Cytopathogenicity assay also revealed that T2 conjugated with AgNPs significantly reduced cytopathogenicity of A. castellanii compared with T2 alone, suggesting that T2-conjugated AgNP is an effective and safe anti-Acanthamoeba agent. The use of a synthetic azole compound conjugated with AgNPs can be an alternative strategy for drug development against A. castellanii. However, mechanistic and in vivo studies are needed to explore further translational values.

Comparative assessment of antifungal potential of Zingiber officinale and Allium sativum against standard antifungal drugs Amphotericin B and Nystatin

ABSTRACT: This study was conducted to investigate theantifungal activity of garlic and ginger on a food derived fungus.Aqueous extracts of garlic (Allium sativum) and ginger (Zingiber officinale) were tested for their antifungal activity and their activities were compared with antifungal drugs nystatin and amphotericin B. The extracts were 40-50% serially diluted and different concentrations were obtained. Sterile strips were dipped in different concentrations of garlic and ginger. Strips were placed on petri plates containing fungus culture. After 24-48 hours incubation at 37 oC, antifungal zones of different diameter were obtained and measured in different plates. The results showed that garlic is more effective in its antifungal activity than ginger. The study demonstrated the potent activity of garlic against tested fungi which encourages its use as a suitable alternative drug for controlling fungal food spoilage. Since garlic is an herbal product with no known side-effects on human healthiest active ingredients may serve as safe food preservatives. This can ultimately increase the shelf life and also maintain the quality of preserved food. Purification and formulation of the garlic would give a true antifungal activity comparable to standard antifungal drugs., , antifungal activity,,

The Substantial Improvement of Amphotericin B Selective Toxicity Upon Modification of Mycosamine with Bulky Substituents.

It is assumed that the unfavorable selective toxicity of an antifungal drug Amphotericin B (AmB) can be improved upon chemical modification of the antibiotic molecule. The aim of this study was verification of the hypothesis that introduction of bulky substituents at the amino sugar moiety of the antibiotic may result in diminishment of mammalian in vitro toxicity of thus prepared AmB derivatives. Twenty-eight derivatives of AmB were obtained upon chemical modification of the amino group of mycosamine residue. This set comprised 10 N-succinimidyl-, 4 N-benzyl-, 5 Nthioureidyl- and 9 N-aminoacyl derivatives. Parameters characterizing biological in vitro activity of novel compounds were determined. All the novel compounds demonstrated lower than AmB antifungal in vitro activity but most of them exhibited negligible cytotoxicity against human erythrocytes and three mammalian cell lines. In consequence, the selective toxicity of majority of novel antifungals, reflected by the selective toxicity index (STI = EH50/IC50) was improved in comparison with that of AmB, especially in the case of 5 compounds. The novel AmB derivatives with the highest STI, induced substantial potassium efflux from Candida albicans cells at concentrations slightly lower than IC50s but did not trigger potassium release from human erythrocytes at concentrations lower than 100 μg/mL. Some of the novel AmB derivatives can be considered promising antifungal drug candidates.

Design and Facile Synthesis of New Bioactive C-glycosidic Semi-Natural Compounds

Objective: The presented work aims to synthesize new bioactive molecules of wide spectrum biological activity. In this study, a new series of α,β unsaturated ketones having acylic sugar residue was synthesized along with their hydrazone analogues. Methods: Those new compounds were obtained via mild condensation reactions to avoid side products and were investigated by spectral analysis including IR, 1HNMR,13CNMR and mass spectrometer. In addition, they were tested and evaluated as antimicrobial and antioxidant agents. Novelty: Results revealed that compound 2d is the most promising antimicrobial agent among the tested compounds and it even exceeds the activity of the used standard antibacterial drug Ampicillin as well as it competes with the standard antifungal drug Amphotericin B. On the other hand, compound 3b shows potent antioxidant activity in comparison with the used standard drug Trolox. Structure-activity relationship is also discussed.

Antifungal activity of silver salts of Keggin-type heteropolyacids against Sporothrix spp.

Sporotrichosis is a chronic and subacute mycosis causing epidemiological outbreaks involving sick cats and humans in southeastern Brazil. The systemic disease prevails in cats, and in humans, the symptoms are restricted to skin in immunocompetent individuals. Under these conditions, the prolonged treatment of animals and cases of recurrence justify the discovery of new treatments for sporotrichosis. This work addresses the antifungal activity of silver salts of Keggin-type heteropolyacid salts (Ag-HPA salts) such as Ag3[PW12O40], Ag6[SiW10V2O40], Ag4[SiW12O40] and Ag3[PMo12O40] and interactions with the antifungal drugs itraconazole (ITC), terbinafine (TBF) and amphotericin B (AMB) on the yeast and mycelia forms of Sporothrix spp. Sporothrix spp. yeast cells were susceptible to Ag-HPA salts at minimum inhibitory concentration (MIC) values ranging from 8 to 128 μg/mL. Interactions between Ag3[PW12O40] and Ag3[PMo12O40] with itraconazole and amphotericin B resulted in higher antifungal activity with a reduction in growth and melanization. Treated cells showed changes in cell membrane integrity, vacuolization, cytoplasm disorder, and membrane detachment. Promising antifungal activity for treating sporotrichosis was observed for the Ag-HPA salts Ag3[PMo12O40] and Ag3[PW12O40], which have a low cost, high yield and activity at low concentrations. However, further evaluation of in vivo tests is still required.

Silymarin, a Popular Dietary Supplement Shows Anti-Candida Activity.

Silymarin is a complex of plant-derived compounds obtained from the seed shells of the milk thistle (Silybum marianum). It is used in medicine primarily to protect the liver. The mixture contains mainly flavonolignans, with silybin as a paramount bioactive component of the extract. This article presents the potential health benefits for silymarin as an antifungal drug against five references strains: C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, and C. krusei with MIC (minimum inhibitory concentration) values ranging from 30 to 300 µg/mL. Additionally, this study revealed that the compound suppressed the growth of cells of most of the tested clinical Candida albicans strains with MIC values between 30 and 1200 µg/mL. Based on the fractional inhibitory concentration index (FICI), the combination of silymarin with antifungal drugs caspofungin, fluconazole, and amphotericin B did not significantly change the MIC values for the tested Candida strains. Furthermore, no antagonistic reactions were observed in any combination of drugs. In addition, this substance shows anti-virulence properties including the destabilization of mature biofilm and the inhibition of the secretion of hydrolases. qRT-PCR-based experiments demonstrated that the SAP4 gene involved in virulence was downregulated by silymarin. These results indicate completely new advantages of dietary supplementation with this natural plant extract.

Candida albicans Infection Model in Zebrafish (Danio rerio) for Screening Anticandidal Drugs.

Candida albicans is an opportunistic fungal pathogen which causes systemic infections in human. In this study, C. albicans infection model was developed in zebrafish to understand the host-pathogen interactions for straightforward anticandidal drug screening. To develop the infection, 1 × 106cells of C. albicans suspended in phosphate-buffered saline were deposited in zebrafish dorsal muscle by manually operated syringe. The infection progression was externally assessed by a scale of wound-healing events, based on visible changes of yeast deposited in the muscle tissues. Chemotherapy was carried out with known antifungal drugs (fluconazole, nystatin, and amphotericin B) and a potential antifungal agent, chitosan silver nanocomposites (CAgNC), after the infection as direct exposure in the water. Histopathological analysis was performed to identify the pathogen virulence and the host-pathogen interaction during the infection. The light microscopic observations and histopathological analysis revealed the yeast-hyphae transition at the site of infection (at 72hpi) and progression of the infection in the host tissues. The larval survival rate under fluconazole (up to 80μgmL-1) and nystatin (up to 20μgmL-1) was > 90% and for CAgNC it was 40% at 36h post-exposure (hpe). The infection progression was suppressed with the fungicidal treatments. Among inflammatory genes, il-1β has been highly upregulated (14.68-fold) at 24h post infection (hpi). Both il-1β and tnf-α were moderately upregulated in infected fish gills at 72hpi. Among the C. albicans antioxidant genes, cat1 and sod2 have been upregulated during the infection, and relative expression folds were increased from low to moderate levels with the time. We demonstrate the approach for the development of artificial infection model of zebrafish with C. albicans. By this mini vertebrate zebrafish model, researchers will be able to study novel anticandidal compounds in vivo with respect to the host, pathogen, and their interactions.

Genomewide Elucidation of Drug Resistance Mechanisms for Systemically Used Antifungal Drugs Amphotericin B, Caspofungin, and Voriconazole in the Budding Yeast.

There are only a few antifungal drugs used systemically in treatment, and invasive fungal infections that are resistant to these drugs are an emerging problem in health care. In this study, we performed a high-copy-number genomic DNA (gDNA) library screening to find and characterize genes that reduce susceptibility to amphotericin B, caspofungin, and voriconazole in Saccharomyces cerevisiae We identified the PDR16 and PMP3 genes for amphotericin B, the RMD9 and SWH1 genes for caspofungin, and the MRS3 and TRI1 genes for voriconazole. The deletion mutants for PDR16 and PMP3 were drug susceptible, but the other mutants had no apparent susceptibility. Quantitative-PCR analyses suggested that the corresponding drugs upregulated expression of the PDR16, PMP3, SWH1, and MRS3 genes. To further characterize these genes, we also profiled the global expression patterns of the cells after treatment with the antifungals and determined the genes and paths that were up- or downregulated. We also cloned Candida albicans homologs of the PDR16, PMP3, MRS3, and TRI1 genes and expressed them in S. cerevisiae Heterologous expression of Candida homologs also provided reduced drug susceptibility to the budding yeast cells. Our analyses suggest the involvement of new genes in antifungal drug resistance.

A novel anti Candida albicans drug screening system based on high-throughput microfluidic chips

Due to the antibacterial resistance crisis, developing new antibacterials is of particular interest. In this study, we combined the antifungal drug amphotericin B with 50,520 different small molecule compounds obtained from the Chinese National Compound Library in an attempt to improve its efficacy against Candida albicans persister cells. To systematically study the antifungal effect of each compound, we utilized custom-designed high-throughput microfluidic chips. Our microfluidic chips contained microchannels ranging from 3 µm to 5 µm in width to allow Candida albicans cells to line up one-by-one to facilitate fluorescence-microscope viewing. After screening, we were left with 10 small molecule compounds that improved the antifungal effects of amphotericin B more than 30% against Candida albicans persister cells.

Antifungal Potential of the Bioactive Constituents in Extracts of the Mostly Untapped Brown Seaweed Hormophysa cuneiformis from The Egyptian Coastal Waters

MYCOSES and plant fungal pathogens are limiting factors highly affecting public health and crop production. Some fungal strains have been documented to be resistant to the commonly used drugs. Therefore, finding out new and pivotal antifungal drugs is becoming a global priority. Herein, we evaluated the in vitro antifungal activities of different crude polar (methanol and ethyl acetate) and non-polar (chloroform and petroleum ether) extracts of the mostly untapped brown seaweed Hormophysa cuneiformis (order Fucales, Phaeophyceae), in the Egyptian coastal waters, against eight pathogenic fungi: Aspergillus flavus, A. fumigatus, Candida albicans and Trichosporonas ahii (as human pathogens), and Alternaria alternata, Cladosporium herbarum, Fusarium oxysporum and Penicillium digitatum (as plant pathogens). The agar well diffusion assay was applied. Our findings showed that the chloroform extract only exhibited a potential antifungal activity against all tested fungal isolates, particularly T. asahii, C. albicans, A. fumigatus and C. herbarum, while the other extracts had relatively no remarkable effects. The minimum inhibitory concentrations (MICs) ranged between 0.78 and 6.25μg.ml-1 and these values are very close to those of the standard antifungal drug amphotericin B (0.63–5μg.ml-1). GC–MS analysis of the crude chloroform extract revealed 45 different bioactive compounds, mainly including 18 different species of saturated, monounsaturated and polyunsaturated fatty acids (71.48%) and some essential oils. The major constituents were arachidonic (C20:4, ω–6; 16.18%), oleic (C18:1, ω–9; 15.61%), palmitic (C16:0; 9.18%) and dihomo-γ-linolenic (C20:3, ω–6; 8.97%) fatty acids.