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Background and Purpose: The present study was targeted toward investigating the effects of eugenol on Cryptococcus neoformans biological activity and Cxt1p gene expression. Materials and Methods: For the purpose of the study, the growth, urease, synergism activity, and disk diffusion of C. neoformans were assessed in eugenol-treated culture. The minimum inhibitory concentration (MIC) was determined by the Clinical and Laboratory Standards Institute M27-A3 method at a concentration range of 0.062-2 mg/mL. Subsequently, the expression of Cxt1p genes was studied at the MIC50 concentration of eugenol using real-time polymerase chain reaction. Results: The obtained results showed that eugenol at the concentrations of 125 and 500 μg/mL resulted in 50% and 100% growth inhibition in C. neoformans, respectively. In terms of urease activity, the results showed that the addition of MIC50 of eugenol and fluconazole to urea medium reduced urease activity in C. neoformans. In the culture treated with eugenol, the inhibition zone of antifungal drugs, namely amphotericin B, itraconazole, and fluconazole, was increased to 36±0.002, 22±0.001, and 12±0.002 mm, respectively. The expression levels of Cxt1p in the eugenol-treated, fluconazole-treated, and non-treated samples were estimated at 46%, 58%, and 100%, respectively. Conclusion: The findings of the current study revealed that eugenol could cause C. neoformans growth inhibition and reduce Cxt1p expression in this species. As the results indicated, the susceptibility of C. neoformans to fluconazole was increased when combined with eugenol.

Background and Purpose: Soil bacteria have extreme population diversity among natural sources and are able to produce a wide array of antifungal metabolites. This study aimed to isolate and identify the bioactive metabolite-producing bacteria from forest soils and evaluate their antimicrobial potent against some pathogenic organisms. Materials and Methods: In this study, soil samples were screened for antifungal activity against Aspergillus fumigatus on glucose-yeast extract (GY) agar using a visual agar plate assay method. All growing bacteria were examined for antifungal activity, and antagonistic bacteria were identified based on 16S ribosomal RNA sequence analysis. For optimization of the production of antifungal bioactive metabolites, inhibitory bacteria were cultured on different culture conditions, including media, pH, temperature, and incubation time. Results: In total, 110 bacterial strains were isolated from the forest soils and four species with high antifungal activity were identified as Streptomyces libani, Streptomyces angustmyceticus, Bacillus subtilis, and Sphingopyxis spp. on the basis of 16s ribosomal RNA sequencing. Dichloromethane extract of the starch casein broth culture filtrate of the S. libani (incubated at 30° C for five days) showed strong antifungal activity against A. fumigatus, Aspergillus niger, and Aspergillus flavus. Conclusion: Based on the results, forest soils contain organisms with antifungal activity and could be considered as a good source for novel antifungal metabolites as effective and safe therapeutics.

Objective Multilocus sequence typing is a powerful method for genotyping of clinical isolates of Candida albicans. Cross-contamination between the patients is an important reason for nosocomial infections. Oropharyngeal candidiasis (OPC) caused by C. albicans is an important problem in patients with head and neck cancer, in Cancer Institute of Tehran. Here we studied the endemic genotypes of C. albicans isolates and the relationship between geographic distributions, potential cross-contaminations and the expression of SAP2 gene, an important gene in oral candidiasis, with MLST groups. Material and methods A total of 32 clinical strains of C. albicans isolated from head and neck cancer patients with oropharyngeal candidiasis were subjected to MLST analysis and SAP2 gene expression was analyzed by Real-time PCR. Results We identified 75 polymorphic sites in 7 loci of C. albicans isolates and 30 diploid sequence types which 27 of them were found as new. After eBURST analysis, our results determined that CC 124 was the most prevalent group among all CCs. SAP2 gene showed high expression in almost all OPC patients’ isolates, compared to the control. Conclusion We found few genetically-related as well as identical isolates among the 32 Candida strains which indicated low cross-contaminations among the patients. There was no relationship between C. albicans MLST profiles and their geographic distribution, cancer type and SAP2 gene expression. This lack of correlation was possibly due to the small understudy population; hence, finding more relevance requires studies with a higher number of samples.

Due to the increasing rate of invasive fungal infections and emerging antifungal resistance, development of novel antifungal drugs has been an urgent necessity. Antifungal peptides (AFPs) have recently attracted attention due to their unique ability to evade drug-resistant fungal pathogens. In this study, a novel AFP, Cc-AFP1, with a molecular weight of ~3.759 kDa, was isolated from Carum carvi L., purified by ammonium sulfate precipitation and reversed-phase HPLC and finally identified by sequence analysis using Edman degradation. Peptide sequence analysis revealed a fragment of 36 amino acid residues as RVCFRPVAPYLGVGVSGAVRDQIGVKLGSVYKGPRG for Cc-AFP1 with a net charge of +5 and a hydrophobicity ratio of 38%. The antifungal activity of Cc-AFP1 was confirmed against Aspergillus species with MIC values in the range of 8–16 µg/ml. Cc-AFP1 had less than 5% hemolytic activity at 8–16 µg/ml on human red blood cells with no obvious cytotoxicity against the HEK293 cell line. Stability analysis showed that the activity of Cc-AFP1 was maintained at different temperatures (20°C to 80°C) and pH (8 to 10). The results of a propidium iodide uptake and transmission electron microscopy showed that the antifungal activity of Cc-AFP1 could be attributed to alteration in the fungal cell membrane permeability. Taken together, these results indicate that Cc-AFP1 may be an attractive molecule to develop as a novel antifungal agent combating fungal infections cause by Aspergillus species.

Introduction: This study aimed to evaluate antifungal activity and cytotoxicity of two new nitroheteroaryl-1,3,4-thiadiazole derivatives containing N-methoxyethyl (9) and N-benzyl (10) moiety against Aspergillus fumigatus with a special focus on their mechanism of action at cellular level. Materials and Methods: The fungal growth rate was evaluated by microbioassay technique. Ergosterol content of the cell membrane was determined, and morphological changes of fungal compartments were assessed by electron microscopy. Cytotoxicity against Vero and Hep2 cell lines was determined by cell culture technique. Results: Based on the data obtained, compound 9 showed 31.90-100% inhibition at concentrations of 25-800 μM, while compound 10 presented 24.26-100% inhibition at higher concentrations (25-3200 μM). Compounds 9 and 10 showed minimum fungicidal concentration of 500 and 8000 μM and IC50 values of 43.39 and 1008.67 μM, respectively. Ergosterol content was not meaningfully affected by both compounds. Electron microscopy showed deformation of A. fumigatus hyphae, depletion of hyphae contents, and destruction of cell membrane and membranous organelles in the fungus exposed to both compounds. Compounds 9 and 10 had no obvious cytotoxicity against Vero and Hep2 cell lines in vitro. Conclusion: Taken together, our results showed that both compounds, especially compound 9, could be considered as potential candidates for developing antifungal drugs against A. fumigatus as the main etiologic agent of life-threatening invasive aspergillosis.

BackgroundDermatophytes are a homogeneous group of species with low genetic diversity, and there are still many uncertainties about the boundaries among species.ObjectivesAiming at clarifying the relationships among species in the genus and introducing suitable genes for multilocus sequence typing (MLST), a new MLST scheme approach was developed to characterize the major pathogenic dermatophytes.MethodsWe performed maximum parsimony (MP), MrBayes, RAxML, and eBURST analyses, based on the MLST scheme to scrutinize the evolution within 95 clinical isolates and four reference strains belonging to the four major dermatophytes species. Then, the discriminatory power, pairwise genetic distances, ratio dN/dS, and sequence types (STs) of these isolates were determined. Also, to study taxonomy, sequences of the internal transcribed spacer (ITS), Beta-tubulin (BT2), and translation elongation factor 1-α (TEF-1α) genes of other dermatophytes species available in the GenBank were analyzed.ResultsFindings of the present study indicated that three genes: BT2, ITS, and TEF−1α, which showed the greatest diversity among dermatophyte species, were suitable for MLST. The most prevalent STs were seen among the species of Trichophyton interdigitale. Also, two new genotypes, i.e., XXVII and XXVIII, were introduced for T. interdigitale and Trichophyton mentagrophytes. The least informative sites were found in Epidermophyton floccosum, Trichophyton rubrum, and T. mentagrophytes, while the most informative sites were observed in T. interdigitale. Furthermore, the most informative locus was TEF-1α. The phylogenetic tree, constructed by the combination of the three genes, shows a new topological pattern that confirms the derivation of the anthropophilic and zoophilic genera from the geophilic genus. Also, the phylogenetic analyses and pairwise distances of the combination of the three loci showed that Trichophyton tonsurans and Trichophyton equinum were a species complex, where T. equinum is derived from T. tonsurans.ConclusionsResults of this study showed that MLST is very effective in determining the boundaries between species and taxonomy. Considering that there is no database for MLST dermatophytes, further studies are needed to determine the suitable genes for MLST. Also, the determination of STs in epidemiological studies and raising epidemiological information are helpful. This study was a new starting point to determine the ST and a foundation for a dermatophyte MLST database.

Background Members of the Pleosporaceae family are known as important sources of airborne allergens which are responsible for asthma and allergic diseases. Aims The purpose of this study was to investigate the gene profiling and expression pattern of Alt a 1 in Alternaria alternata and other members of the Pleosporaceae family including Stemphylium botryosum, Ulocladium chartarum, Curvularia lunata, Cladosporium cladosporioides, and Epicoccum nigrum. Methods Alternaria alternata and related genera were cultured on Czapek–Dox broth medium at 25 °C for 21 days. The presence of Alt a 1 was assessed in fungal culture filtrates by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS–PAGE) and then confirmed by immunoblot analysis. Real-time PCR was carried out for quantitation of the Alt a 1 gene encoding corresponding protein at the transcriptional level using cDNA prepared from fungal RNA. Results SDS–PAGE showed protein bands ranging from 14 to 100 kDa. A 14 kDa band corresponding to Alt a 1 was present in A. alternata, S. botryosum and U. chartarum. The gene expression of Alt a 1 was reported in A. alternata and some other related genera. The Ct mean value recorded for A. alternata strains ranged from 24.70 to 27.84 while it was in the range 23.62–32.09 for other related taxa. No apparent transcription or expression was revealed in C. cladosporioides. Conclusions The presence and efficient expression of Alt a 1 gene in A. alternata and other related taxa indicate that Alt a 1 protein is a major component of the secretory machinery of Pleosporaceae family members, and it may play a crucial role in its allergenicity.

Ubiquitous airborne conidia of the genus Aspergillus are responsible for a diverse group of human disorders from allergy to life treating invasive aspergillosis and mycotoxicoses. The aim of this study was to determine the population structure of Aspergillus isolated from outdoor air in Tehran by comparing the nucleotide sequences of ITS region and the PCR-RFLP molecular method. Internal transcribed spacer domains of 47 Aspergillus spp. were amplified and sequenced and PCR products were digested individually with restriction enzymes TaqI and EcoRI. For all species the PCR reaction produced a fragment of approximately 600 bp in length. All of the nucleotide sequences were highly similar with the corresponding reference sequences registered at the gene bank. The all isolates displayed same banding pattern on the basis EcoR1 cleavage. While Taq1 enzyme profiling provided 5 different banding pattern. The results show that the A. niger section has the highest frequency with 27 isolates (57.4%). Of these, 23 isolates (48.9%) belonged to the A. niger complex and 4 isolates (8.5%) to the A. aculeatus complex. The A. flavus complex was also placed in the next ranking with 9 isolates (19.1%). These results strongly support the need for using molecular markers as an auxiliary tool in differentiating Aspergillus species.

Background and Purpose: This study aimed to investigate the effects of Allium cepa ethanolic extract (EAC) on Cryptococcus neoformans biological activities and LAC1 gene expression. Materials and Methods: The minimum inhibitory concentration (MIC) of EAC was determined based on the Clinical and Laboratory Standards Institute M27-A4 method at a concentration range of 125- 4000 µg/ml. The EAC synergism activity was determined in combination with fluconazole (FCZ) as an antifungal azole. Laccase activity, melanin production, and cell membrane ergosterol content of C. neoformans were assessed at the 0.5× MIC concentration of EAC (1000 μg/ml) and FCZ (64μg/ml) by approved methods. The expression of the LAC1 gene was studied in the fungus exposed to 0.5× MIC concentration of EAC and FCZ using the real-time polymerase chain reaction. Results: Based on obtained results, MIC of EAC and FCZ were 2000 and 128 μg/ml,respectively. A combinatory effect was reported for FCZ and EAC by a fractional inhibitory concentration index of 0.25. The cell membrane ergosterol content was inhibited in EAC- and FCZ-treated C. neoformans by 58.25% and 49.85%, respectively.The laccase activity and melanin production were reduced in EAC-treated C. neoformans by 45.37% and 51.57%, and in FCZ-treated fungus by 54.64% and 53.68%, respectively.The expression of fungal LAC1 at messenger RNA (mRNA) level was measured 0.46 and 0.58 folds and significantly decreased in both EAC- and FCZ-treated C. neoformans at the 0.5×MIC concentration, respectively (p

Aims This study aimed to investigate the mechanism of antifungal action of Streptomyces libani dichloromethane extract fraction A (DCEFA) against Aspergillus fumigatus and the host cytotoxicity. Methods and results DCEFA was purified from S. libani by autobiography and showed strong antifungal activity against A. fumigatus. A combination of electron microscopy, cell permeability assays, total oxidant status (TOS) assay, cell cytotoxicity assay, and hemolysis activity were carried out to determine the target site of DCEFA. Exposure of A. fumigatus to DCEFA caused the damage to membranous cellular structures and increased release of cellular materials, potassium ions and TOS production. DCEFA was bound to ergosterol but did not affect fungal cell wall and ergosterol content. DCEFA did not show any obvious hemolytic activity for RBCs and toxicity against HEK-293 cell line. Conclusions DCEFA may inhibit A. fumigatus growth by targeting fungal cell membrane which results in the leakage of potassium ions and other cellular components, TOS production and final cell death. Significance and Impact of the Study DCEFA of S. libani could be considered as a potential source of novel antifungals which may be useful for drug development against Aspergillus fumigatus as a life-threatening human pathogen.

Background The Candida albicans is one of the most important global opportunistic pathogens, and the incidence of candidiasis has increased over the past few decades. Despite the established role of skin in defense against fungal invasion, little has been documented about the pathogenesis of Candida species when changing from normal flora to pathogens of vaginal and gastrointestinal epithelia. This study was carried out to determine the in vivo and in vitro pathogenesis of clinical C. albicans strains isolated from skin lesions. Methods In this study, association of in vivo and in vitro pathogenesis of C. albicans isolates with different evolutionary origins was investigated. Oral and systemic experimental candidiasis was established in BALB/C mice. The expression levels of secreted aspartyl proteinases (SAP1-3 genes), morphological transformation, and biofilm-forming ability of C. albicans were evaluated. Results All the strains showed in vitro and in vivo pathogenicity by various extents. The SAP1, SAP2, and SAP3 genes were expressed in 50%, 100%, and 75% of the strains, respectively. The biofilm formation ability was negative in 12% of the strains, while it was considerable in 38% of the strains. Fifty percent of the strains had no phospholipase activity, and no one demonstrated high level of this pathogenesis factor. Relatively all the strains had very low potency to form pseudohyphae. Conclusion Our findings demonstrated that Candida albicans strains isolated from cutaneous candidiasis were able to cause oral and systemic infections in mice, so they could be considered as the potential agents of life-threatening nosocomial candidiasis in susceptible populations.

Background: It is estimated that more than 80% of cases of human infections are related to biofilm formation by invasive bacteria. So, in this study, we considered the activity of cellobiose dehydrogenase enzyme (CDH) isolated from Aspergillus niger, as an antibiofilm agent, on biofilm of clinical Staphylococcus epidermidis and Pseudomonas aeruginosa isolates. Methods: In this study, five standard strains of Aspergillus niger were purchased for CDH production. Of the 42 isolated bacterial strains, 24 strains were Staphylococcus epidermidis and 18 strains were Pseudomonas aeruginosa. Zymogram method was used for screening of CDH. The CDH activity was measured by monitoring the decrease in absorbance of 2, 6-dichlorophenolindophenol (DCPIP) spectrophotometrically. After cultivation and production of bacterial biofilms, 7 concentrations of the cellobiose prepared and CDH enzyme with a final concentration of 364 U/mL were considered on bacterial biofilms by microbroth dilution method. Results: Out of five standard strains of Aspergillus niger, only 1 strain have the highest production of CDH. The most effective dilution of cellobiose on growth inhibition of Staphylococcus epidermidis and Pseudomonas aeruginosa in liquid cultures as a function of cellobiose concentration in the presence of cellobiose dehydrogenase was in 12.5 µg/mL. Conclusions: Based on the results of this study, it can be concluded that the CDH enzyme had a high potential for use as an antimicrobial agent. As shown, this enzyme had a high potential for eliminating bacterial biofilms. So, these results may provide a basis for alternative therapies for the treatment of infections related to clinical biofilm producing bacteria.

Candida species are the commensal organisms of human and animal mucosa that cause a wide range of debilitating diseases in immunocompromised patients and other susceptible individuals. The present study aimed to investigate the ability of clinical isolates of various Candida species to produce proteinase and phospholipase, hydrophobicity and biofilm forming ability that assumed to have a vital role in Candida pathogenicity. Eighty-four Candida strains belonged to Candida albicans (44.1%), C. glabrata (5.9%), C. guilliermondii (5.9%), C. krusei (10.8%), C. parapsilosis (26.2%), and C. tropicalis (7.1%) were examined for proteinase and phospholipase production, cell surface hydrophobicity and biofilm forming ability. The production of proteinase and phospholipase was detected in 81 (96.4%) and 79 (94.1%) of the strains, respectively. C. albicans showed the highest proteinase and phospholipase activity (mean Pz values of 0.42 ± 0.25 and 0.72 ± 0.28) and biofilm formation ability (0.66 ± 0.22). C. parapsilosis had the highest hydrophobicity (42.97 ± 16.1), which showed a good correlation with biofilm formation ability. A considerable percentage of non-albicans Candida strains produced significant amounts of proteinase and phospholipase with a good ability of biofilm formation in vitro. Taken together, our results further substantiated that enzymatic activity, hydrophobicity and the ability for biofilm formation are important virulence factors which may be account for pathogenicity of various Candida species distributed in albicans and non-albicans groups.

Background Eugenol, as the major phenolic component of clove essential oil due to its desired properties in medical field, was loaded into polyacrylonitrile (PAN) nanofibers with various percentages. Objective Our main purpose in this study was to determine the in vitro antifungal activity of eugenol loaded on PAN nanofibers against Candida albicans as the most common causative agent for candidiasis. Method Also, the surface morphology and the mechanical properties of nanofibers were studied by scanning electron microscope (SEM) and a tensile tester, respectively. The average diameters of nanofibers in pure PAN nanofibers were found to be 127 nm. Results The results showed that the average diameter of nanofibers after increasing the eugenol ratio (from 127 to 179-218 nm) was increased. Drug release profile of the samples was gradual and was completed after 150 hours. Conclusion According to the results, these nanofiber mats loaded with eugenol can be used for treating cutaneous mucocutaneous candidiasis in high risk patients as a coating on a fabric substrate or temporary wound dressing.

Background Candida glabrata is a yeast that can cause hazardous fungal infections with high mortality and drug resistance. Aims The aim of this study was to determine the profile of drug susceptibility in clinical isolates of C. glabrata and review the resistance mechanisms to caspofungin. Methods A total of 50 C. glabrata clinical isolates from Iran were tested for in vitro susceptibilities to amphotericin B, caspofungin, fluconazole and voriconazole. To investigate the mechanism of resistance to caspofungin, hotspot areas of FKS1 and FKS2 genes were sequenced and gene expression profile was evaluated. Results All the isolates were susceptible to amphotericin B and caspofungin. Fluconazole resistance was exhibited in four isolates. In addition, only one isolate was resistant to voriconazole. FKS2 with 12 point mutations showed more mutations compared to FKS1 that had only two mutations. All substitutions were synonymous. FKS genes were expressed at comparable levels (no statistical significance) in caspofungin-treated and non-treated cultures. Conclusions The silent mutations in the hotspot areas of FKS genes and inconsiderable changes in gene expression were not associated with increased MIC (0.25 μg/ml). Other mechanisms of resistance which include mutations outside the hotspot area of FKS genes could be involved in a slight increase of MIC, and they should be identified through complete FKS gene sequencing.

In the present study, halophilic bacteria communities were explored in saline soils of Howze-Soltan playa in Iran with special attention to their biological activity against an aflatoxigenic Aspergillus parasiticus NRRL 2999. Halophilic bacteria were isolated from a total of 20 saline soils using specific culture media and identified by 16S rRNA sequencing in neighbor-joining tree analysis. Antifungal and antiaflatoxigenic activities of the bacteria were screened by a nor-mutant A. parasiticus NRRL 2999 using visual agar plate assay and confirmed by high-performance liquid chromatography. Among a total of 177 halophilic bacteria belonging to 11 genera, 121 isolates (68.3%) inhibited A. parasiticus growth and/or aflatoxin production. The most potent inhibitory bacteria of the genera Bacillus, Paenibacillus and Staphylococcus were distributed in three main phylogenetic clusters as evidenced by 16S rRNA sequence analysis. A. parasiticus growth was inhibited by 0.7–92.7%, while AFB1 and AFG1 productions were suppressed by 15.1–98.9 and 57.0–99.6%, respectively. Taken together, halophilic bacteria identified in this study may be considered as potential sources of novel bioactive metabolites as well as promising candidates to develop new biocontrol agents for managing toxigenic fungi growth and subsequent aflatoxin contamination of food and feed in practice.

Pneumocystis jirovecii is an opportunistic fungus, which causes Pneumocystis pneumonia (PJP) in immunocompromised, COPD and TB positive patients with a high rate of colonization, morbidity and mortality. Dihydropteroate synthase (DHPS) gene mutations are well-reported in PJP. Although sulfa prophylaxis generally is associated with DHPS mutant infection, Multiple molecular techniques applied for detect sulfa resistance single-base mutation. Conformation sensitive gel electrophoresis (CSGE) is a rapid screening method for detection of DNA sequence variation, specifically single-base changes or small insertions and deletions. The current study is investigate on the DHPS sequence single-base dislocation among strains isolated from Iranian TB positive co-infected with PJP in association to increased levels of serum Lactate Dehydrogenase. Through high serum lactate dehydrogenase (LDH) levels have been associated with established Pneumocystis pneumonia. We investigated the DHPS mismatches in five P. jirovecii isolated of TB infected patients. For genetic identification of Pneumocystis isolates and detection of intraspecific variation, we developed a method for heteroduplex analysis. Our utilizing fragments was the DHPS gene regions, amplified by PCR method with specific primers. Serum LDH indicator was analysed for lung acute damages. In our results, at least 4 suspected isolates show more slowly migrating bands containing single heteroduplexes, reveal single-base mislocation in studied sequences. LDH level Peak was higher (p

In spite of developing new drugs and modern formulations, the treatments of chronic fungal infections are still challenging. Fibrous wound dressings are new suggestions for the treatment of chronic superficial infections. In the present study, we formulated an antifungal agent, terbinafine hydrochloride (TFH), which is a hydrophobic drug, in wound dressings prepared by electrospun polycaprolactone, polycaprolactone/gelatin (50:50 w/w) and gelatin. To obtain more water-stable meshes, the preparations were treated by glutaraldehyde and their properties were determined before and after treatment. The morphology of fibrous meshes was observed by scanning electron microscopy. Drug loading efficiency and release rate were measured by high performance liquid chromatography (HPLC) and the release rate was monitored for 144h. Antifungal tests were performed on Trichophyton mentagrophytes, Aspergillus fumigatus and Candida albicans cultured on Muller-Hinton agar. The toxicity of the meshes was measured after 24h and 14days by MTT assay. Terbinafine loading of polycaprolactone/gelatin (50:50) was 100% and it released the highest amount of TFH too. In antifungal tests, all samples were able to hinderT. mentagrophytes and A. fumigatus but not C. albicans growth among them, polycaprolactone fibers made the largest inhibition zone. In MTT assay, none of prepared samples showed toxicity against L929 cells. Teken together, the prepared TFH-loaded PCL/gelatin electrospun meshes were able to release TFH slowly and in a steady state in time. With respect to no obvious cytotoxicity in MTT assay and stong antifungal activity toward T. mentagrophytesin vitro, these TFH-based meshes could be considered as potential candidates in clinical application as wound dressing for treatment of chronic dermatophytosis.

Summary Objective Dermatophytosis is the most frequent fungal infection all over the world and its frequency is constantly increasing. The aim of this study was to evaluate clinical features and epidemiological trends of dermatophytosis over the years 2010 to 2014 in Tehran, Iran. Patients and methods A total of 13,312 patients clinically suspected of cutaneous fungal infections were examined. Skin scales, plucked hairs, nail clippings and sub-ungual debris were examined by direct microscopy and culture. Dermatophyte species were identified at the species level by a combination of morphological and physiological criteria. Results Direct microscopy confirmed a contamination rate of 19.7% (2622/13,312 cases) of which 1535 cases (58.5%) were culture positive distributed in male (1022 cases) and female (513 cases). The most commonly infected age group was the 30–39 years old. Tinea pedis (30.4%) was the most prevalent type of dermatophytosis followed by tinea cruris (29.8%) and tinea corporis (15.8%). Epidermophyton floccosum (31%) was the most prevalent causative agent, followed by Trichophyton rubrum (26.2%) and Trichophyton mentagrophytes (20.3%). Conclusion Our results showed considerable distribution of dermatophytosis from zoophilic, anthropophilic and geophilic species among population with diverse age groups. Although anthropophilic fungi such as T. mentagrophytes , E. floccosum , and T. rubrum were the main etiologic agents of dermatophytosis, the prevalence of T. verrucosum showed a meaningful increase over the years, which highlights the importance of rural dermatophytosis mainly transmitted from large animals. This noticeable information improves our current knowledge about dermatophytosis and assists to establish effective prevention and therapeutic strategies to overcome the disease.

Background Dermatophytosis is the most important superficial fungal infection which affects nearly 20% of human population worldwide. Recurrence of disease and emerging resistance of Trichophyton rubrum to synthetic antifungals are the main problems in control of dermatophytosis. The purpose of this study was to evaluate the effect of cold atmospheric plasma (CAP) on T. rubrum growth, ergosterol biosynthesis and keratinase activity. Methods A CAP system, comprised of helium 98% – oxygen 2% (He/O2), was used. Trichophyton rubrum conidia suspensions were treated with CAP in time periods of 90, 120, 150 and 180 s in 96-well microplates. Fungal growth was evaluated by counting the colony forming unit (CFU). Fungal dry weight, ergosterol biosynthesis and keratinase activity were evaluated in CAP-treated T. rubrum and untreated controls. Results T. rubrum growth was significantly inhibited by 62%–91%. CAP strongly suppressed fungal ergosterol biosynthesis by 27%–54%. The keratinase activity was increased by 7.30%–21.88% up to 120 s CAP exposure. Conclusion Our results demonstrated for the first time that CAP inhibits T. rubrum growth, suppresses ergosterol biosynthesis and increases moderately keratinase activity in a dose-dependent manner. Overall, CAP exposure could be a potentially useful method for treatment of clinical cases of human and animal dermatophytoses.

The development of efficient cell-free systems of nanoparticle synthesis using microbial enzymes is a growing field of biological and green chemistry for the supportable improvement in nano-biotechnology. In the present study, we established a cell-free system for producing gold nanoparticles (AuNPs) using a fungal oxidoreductase named sulfite oxidoreductase purified to homogeneity from Fusarium oxysporum. The enzyme was purified by ultrafiltration followed by anion exchange chromatography on DEAE Sephadex A-50 gel, and its molecular weight was determined by gel filtration chromatography on Sephacryl S-300 gel. The purified enzyme had a molecular weight of 346 kDa. It was composed of three subunits of 176, 94 and 76 kDa. Purified enzyme was successfully used for production of gold nanoparticles in a cell-free system. Synthesized gold nanoparticles showed the highest absorbance at 520 nm wavelength as shown by UV–visible spectroscopy. They were spherical in shape with an average size of 20 nm as determined by scanning and transmission electron microscopy and dynamic light scattering. Assessment of the antifungal properties of synthesized nanoparticles by disk diffusion method indicated a potent growth inhibitory activity against all tested human pathogenic yeasts and molds by inhibition zones ranged from 10 to 18 mm. Taken together, our enzymatically established method of nanoparticle synthesis using a purified sulfite oxidoreductase of F. oxysporum can be considered as an efficient tool for generating harmless bioactive gold nanoparticles with potential applications in biology, medicine and industry.

Background The pathogenic yeast Candida albicans is the most common opportunistic fungal pathogen that is responsible for a wide array of infections in susceptible individuals. Despite recent progress in developing novel antifungal drugs which combat Candida -related disorders, this fungus is still a major cause of life-threatening infections all over the world. In the present study, the effect of cold atmospheric plasma (CAP) was evaluated on the growth of C. albicans with special attention to the ability of the CAP-treated fungus for biofilm formation, ergosterol biosynthesis and phospholipase and proteinase secretory production. Methods C. albicans cell suspensions were irradiated over time-scales ranging of 90, 120, 150 and 180 s under cold atmospheric plasma contained He/O 2 (2%). Treated and untreated yeast cells were analyzed for the growth, biofilm formation, ergosterol content, and activities of phospholipase and proteinase. Results Our results showed that CAP remarkably suppressed the growth of C. albicans by 31–82% at the given times. Likewise, CAP strongly inhibited the ergosterol biosynthesis by the fungus in the range of 40–91%, biofilm formation by 43–57% and the activities of phospholipase and proteinase enzymes by 4–45%, dose-dependently. Conclusion CAP strongly inhibits the growth and virulence factors of C. albicans and thus, it could be a potential candidate to treat Candida -related superficial and cutaneous infections in practice.

With regard to increasing number of antifungal-resistant dermatophytes, antifungal susceptibility testing of dermatophytes serves as a useful tool in managing clinical dermatophytosis. This study aimed to determine antifungal susceptibility profile of clinically important dermatophytes and determination of point mutations in terbinafine-resistant isolates. Based on our results, dermatophytosis was confirmed in 97 cases by direct microscopic examination, culture, and sequencing of ITS region. Antifungal susceptibility of 97 dermatophyte isolates distributed in four species including Trichophyton interdigitale (26 isolates), T. rubrum (19 isolates), T. tonsurans (29 isolates), and Epidermophyton floccosum (21 isolates) was assessed to nine antifungal agents using CLSI M38-A2 guidelines. Minimum inhibitory concentration range (MIC range) for luliconazole and terbinafine was 0.001-0.008 μg/ml and 0.003-> 32 μg/ml, compared to 0.03-64 μg/ml for griseofulvin, 0.01-16 μg/ml for itraconazole and voriconazole, 0.03-8 μg/ml for ketoconazole, 0.03-32 μg/ml for econazole, 0.03-1 μg/ml for lanoconazole, and 0.01-4 μg/ml for butenafine. Trichophyton tonsurans was the most susceptible (MIC = 0.006 μg/ml) and E. floccosum was the most resistant (MIC = 0.02 μg/ml) species to terbinafine. Terbinafine resistance was reported for two species, i.e., T. rubrum and T. tonsurans at the total rate of 2% which was due to Leu393Phe substitution in both species. Taken together, our results assist clinicians and prompt the current knowledge about the necessity of antifungal susceptibility testing to select effective strategies for management of clinical cases of dermatophytosis.

Some filamentous fungi are plant pathogens that cause serious economic losses in foodstuffs and agricultural commodities. In the present study, the antifungal effects of culture filtrates of soil strains of Bacillus subtilis and Bacillus amyloliquefaciens were studied against some common field-pathogenic fungi with special reference to the possible mode of actions against Aspergillus parasiticus NRRL2999. The fungal growth and aflatoxin production were determined by microbioassay technique. Changes in membrane ergosterol content, mitochondrial dehydrogenase activity, and chitin and β-1,3-glucan content of the fungal cell wall were evaluated as the different possible targets in the presence of various concentrations of bacterial culture filtrates. According to the results, both bacteria showed strong antifungal activity against the broad spectrum of filamentous fungi. They were able to suppress A. parasiticus growth (up to 92%) and aflatoxin production (up to 100%) and showed good proteolytic activities (up to 10.5 ± 0.4 mm clear zones on gelatin agar). Both antagonistic bacteria decreased ergosterol content of the fungal cell membrane of A. parasiticus (9.0–80%) in a dose-dependent manner. They also affected dehydrogenase activity of fungal cell mitochondria. Interestingly, by using 2500 μl of bacterial culture filtrate, the slight resume of fungal growth and cell wall compositions, referred as the paradoxical effect, was exhibited. Scanning and transmission electron microscopy showed drastic changes in structure and shapeof the treated fungal hyphae and vesicles including folding, wrinkling, cell depletion and vacuolization. Altogether, our results show B. subtilis and B. amyloliquefaciens target different sites in the fungal cells and thus, they could act as biological control agents to combat A. parasiticus growth and subsequent aflatoxin contamination of crops and agricultural commodities.

While the majority of infections in the central nervous system (CNS) are caused by bacteria and viruses, fungi are increasingly being recognized as important pathogens. This is mainly due to the increased awareness of clinicians about these conditions, widespread use of antibacterial agents, a rapid increase in the number of the immunocompromised population, advances in imaging, and the availability of microbiological techniques to confirm the diagnosis from bodily fluids and specimens. Fungal infections have been recognized since early times, but fungal infections of the CNS have mainly been recognized since the 19th century. Fungal infections of the CNS include a wide spectrum of clinical syndromes comprising abscesses, meningitis/meningoencephalitis, focal masses, stroke/vasculitides, immune reconstitution inflammatory syndrome (IRIS), and spinal pathologies such as arachnoiditis. The main etiologies comprise Aspergillus, Cryptococcus, Candida, Mucorales, dematiaceous molds, and dimorphic endemic fungi, with the route of acquisition being respiratory or traumatic inoculation with subsequent spread hematogenously or contiguously. Proper management focuses on early effective antifungal therapy and surgery for large or compressive mass lesions. This chapter highlights recent advances in CNS fungal infections caused by different groups of fungal pathogens from etiology to diagnosis and management.

Summary Background We aimed to evaluate the efficacy of alternative therapies rather than the current antifungal conventional therapy and with assessing the hypothesis of photoactivation of citrus essential oil, fluconazole and Indocyanine green to treat two common mucocutaneous fungal infections. Methods Suspensions of Candida albicans and Tricophyton rubrum containing 106 cells/ml was prepared. Equal samples were treated with infrared (IR) laser irradiation (810 nm, 55 J/cm2) in the presence of Indocyanine green (Emundo, 1 mg/ml) (IRLE), photoactivated Citrus aurantifolia essential oil (EO) with sequential exposure to natural and tungsten lights (CE), control non-activated essential oil (CC), laser alone (IRL), indocyanine green alone (E) and neither of treatments as the control group (C). Additional fluconazole (FL, 25.6 μg/ml) and IR activated fluconazole (IRLFL) groups were designed for T. rubrum fungi. Inoculums were serially diluted to 10−2 and 10−4 and streaked on Sabouraud dextrose agar plates. Final outcomes were assessed as the percent of reduction. Results Cell reduction rates (%) in C. albicans groups were 99.99 (CE), 91.67 (IRLE), 86.67 (CC), 72.37 (E) and 67.27 (RL). Whereas, a 99.99 (CE), 89.99 (CC), 74.5 (IRLE), 64.5 (E), 38.5 (IRLF), 37.5 (RL), and 31 (FL) percent eradication was achieved in T. rubrum groups. Conclusion Photoactivation of Citrus EO increased the killing capability by 10–13%. A modest 7.5% augmented effect was observed with IR activation of Fluconazole. Both Citrus EO and photothermal-photodynamic therapy with ICG and IR diode laser exhibited remarkable lethal effect on fungal cells. Candida viable cells are more susceptible to laser only and ICG only treatments than Tricophyton cells.

Background Contamination of food and agricultural crops by Fusarium species is a major concern of food spoilage and a potential public health hazard. In the present study, natural contamination of maize and wheat samples from main cultivation areas of Iran by Fusarium species belonging to the Giberella fujikuroi species complex was evaluated, with special attention to the ability of the isolates to produce fumonisin B1 (FB1 ). Results A total of 55 Fusarium isolates were obtained from 27/32 maize samples (84.4%) and 11/15 wheat samples (73.3%). They were identified as F. verticillioides (47.3%), F. proliferatum (47.3%), F. fujikuroi (1.8%), F. nygamai (1.8%) and F. redolens (1.8%) by sequence analysis of translation elongation factor 1-α (TEF1-α). Twenty-two of 55 Fusarium isolates belonging to F. proliferatum (23.6%), F. verticillioides (14.5%) and F. fujikuroi (1.8%) produced FB1 in the concentration range 230.4-9565.0 µg mL(-1) . The dendrogram resulting from the TEF1-α profile showed that the genotypes were divided into clusters I, II and III, of which cluster III contained only F. redolens, its first report from Iran. Conclusion On the basis of in vitro FB1 biosynthesis of the analyzed strains, the high degree of contamination of maize and wheat with Fusarium strains reported here should be considered as a potential public health threat, because a meaningful number of the isolates were found to produce hazardous levels of carcinogenic FB1 .

Summary Objective Non-Candida albicans (NCA) species now account for a significant part of clinical candidiasis worldwide. In the present study, epidemiology and antifungal susceptibility profile of NCA isolated from various forms of candidiasis were studied with special focus on their phylogenetic relationship by ITS sequencing. Patients and methods Seventy-nine NCA isolates were isolated from skin and nail scrapings (67.0%), vaginal discharges (8.8%), blood (8.8%), sputa (5.0%), urine (5.0%), oral swabs (2.6%), biopsy and eye tumor, each (1.4%). These isolates were identified by morphological, biochemical and molecular (ITS sequencing) techniques. In vitro antifungal susceptibility of the isolates to fluconazole (FCZ) was tested according to the CLSI method (M27-S4). Results Among a total number of 79 cases of proven NCA infections, C. parapsilosis (36.8%) was the most prevalent species followed by C. glabrata (32.9%), C. orthopsilosis (11.4%), C. tropicalis (8.9%), C. krusei (5.0%) and C. guilliermondii (5.0%). The susceptibility to FCZ was assessed for C. parapsilosis (96.5%), C. orthopsilosis (88.9%), C. tropicalis (85.7%) and C. guilliermondii (50.0%). C. glabrata and C. krusei isolates were not susceptible to FCZ. NCA species were distributed in various phylogenetic clades including C. glabrata (1), C. tropicalis (3), C. parapsilosis (6) and C. orthopsilosis, C. krusei and C. guilliermondii (each 2). Conclusion C. parapsilosis and C. glabrata were the most predominant NCA species involve in the etiology of candidiasis. C. orthopsilosis was reported from superficial candidiasis. Taken together, our results further substantiate the increasing importance of the involvement of NCA species in the etiology of candidiasis.

Finding new compounds with antifungal properties is an important task due to the side effects of common antifungal drugs and emerging antifungal resistance in fungal strains. ∆24-sterol methyltransferase (24-SMT) is a crucial enzyme that plays important roles in fungal ergosterol biosynthesis pathway and is not found in humans. In the present study, the effects of α-bisabolol on Aspergillus fumigatus Af239 growth and ergosterol synthesis on the base of 24-SMT enzyme activity were studied; in addition, the expression of erg6, the gene encoded 24-SMT, was considered. To our knowledge, this is the first report demonstrating that α-bisabolol inhibits A. fumigatus growth specifically via suppressing fungal 24-SMT. Since this enzyme is a specific fungal enzyme not reported to exist in mammalian cells, α-bisabolol may serve as a lead compound in the development of new antifungal drugs. Fungi were cultured in presence of serial concentrations of α-bisabolol (0.281-9 mM) for 3 days at 35 °C. Mycelia dry weight was determined as an index of fungal growth and ergosterol content was assessed. Microsomal 24-SMT activity was assayed in presence of α-bisabolol as an inhibitor, lanosterol as a substrate and [methyl-H3] AdoMet (S-Adenosyl methionin). In addition, the expression of erg 6 gene (24-SMT encoding gene) was determined after treatments with various concentrations of α-bisabolol. Our results demonstrated that α-bisabolol strongly inhibited A. fumigatus growth (35.53-77.17%) and ergosterol synthesis (26.31-73.77%) dose-dependently and suppressed the expression of erg 6 gene by 76.14% at the highest concentration of 9 mM. α-bisabolol inhibited the activity of 24-SMT by 99% at the concentration of 5 mM. Taken together, these results provides an evidence for the first time that α-bisabolol inhibits A. fumigatus Af239 growth via affecting microsomal ∆24-sterol methyltransferase as a crucial enzyme in ergosterol biosynthetic pathway.

Nanostructures from natural sources have received major attention due to wide array of biological activities and less toxicity for humans, animals, and the environment. In the present study, silver nanoparticles were successfully synthesized using a fungal nitrate reductase, and their biological activity was assessed against human pathogenic fungi and bacteria. The enzyme was isolated from Fusarium oxysporum IRAN 31C after culturing on malt extract-glucose-yeast extract-peptone (MGYP) medium. The enzyme was purified by a combination of ultrafiltration and ion exchange chromatography on DEAE Sephadex and its molecular weight was estimated by gel filtration on Sephacryl S-300. The purified enzyme had a maximum yield of 50.84 % with a final purification of 70 folds. With a molecular weight of 214 KDa, it is composed of three subunits of 125, 60, and 25 KDa. The purified enzyme was successfully used for synthesis of silver nanoparticles in a way dependent upon NADPH using gelatin as a capping agent. The synthesized silver nanoparticles were characterized by X-ray diffraction, dynamic light scattering spectroscopy, and transmission and scanning electron microscopy. These stable nonaggregating nanoparticles were spherical in shape with an average size of 50 nm and a zeta potential of -34.3. Evaluation of the antimicrobial effects of synthesized nanoparticles by disk diffusion method showed strong growth inhibitory activity against all tested human pathogenic fungi and bacteria as evident from inhibition zones that ranged from 14 to 25 mm. Successful green synthesis of biologically active silver nanoparticles by a nitrate reductase from F. oxysporum in the present work not only reduces laborious downstream steps such as purification of nanoparticle from interfering cellular components, but also provides a constant source of safe biologically-active nanomaterials with potential application in agriculture and medicine.

The ethyl acetate extract of leaves, seeds and flowers of Heracleum persicum, a medicinal plant of Iran (family Apiaceae) inhibited growth and aflatoxin (AF) production of Aspergillus parasiticus. On the basis of total dry weight growth inhibition by the leaf extract ranged from 17.1 to 36.9 %, by the flower extract from 32.2 to 75.6 %, and by the seed extract from 27.5 to 74.9 %. Production of AFB1 and AFG1 was inhibited in a dose-dependent manner, with a reduction of 88.5-100 % at the highest concentration of 8,000 μg/ml tested. The flower extract decreased ergosterol content of hyphae most significantly. Electron microscopy further revealed structural defects in the treated A. parasiticus including disruption of cytoplasmic membranous compartments, detachment of plasma membrane from the cell wall, and disorganization of hyphal compartments. Collapsed hyphae without conidiation, shorter branches and undifferentiated hyphal tips were also evident. The results indicate that H. persicum extract exerts antifungal and anti-AF activities by disrupting plasma membrane integrity and permeability mainly through interference with ergosterol biosynthesis. These results show that H. persicum can serve as a potent and safe alternative for inhibiting toxigenic aspergilli growth and thus preventing AF contamination of foods and feeds.

Candida glabrata is often the second most common causative agent for candidiasis following Candida albicans. Despite the importance of C. glabrata infections, few epidemiological studies have been conducted on this issue. The goal of this study was genotyping of clinical isolates of C. glabrata by multilocus sequence typing (MLST) technique for determination of the endemic prevalent genotypes and any association between isolation source and drug resistance. A total of 50 C. glabrata clinical isolates from Iran were analyzed by MLST and tested for in-vitro susceptibilities to amphotericin-B, caspofungin, fluconazole, and voriconazole according to the Clinical Laboratory Standards Institute (CLSI) M27-A4 document guidelines. Among these isolates, 16 distinct STs were identified, indicating a discriminatory power index of 0.9029. The three major sequence types (STs) were ST-59, ST-74, and ST-7 with 10, 8, and 7 isolates, respectively. Furthermore, a total of 11 new sequences were found, to which no allele numbers were assigned in the MLST database. All the isolates were susceptible to amphotericin B and caspofungin. Fluconazole resistance was shown in four isolates. Also, a sole isolate was voriconazole resistant. This study shows that the population structure of C. glabrata in Iran consists of groups closely related to the global database as well as to some new clonal clusters and STs. Regarding the high prevalence of 11 new sequences found in this study, it can be concluded that, these new alleles are among the endemic genotypes of Iran. The genotypes or STs were independent of drug susceptibility and anatomic sources.

Summary Objective Dermatomycoses are considered as an important group of fungal diseases with worldwide distribution. This study was performed to analyze the spectrum of dermatomycoses in Tehran during a 4-year period from 2006 to 2009. Patients and methods A total of 12461 patients were investigated for mycoses. Of these, 4871 cases were positive. Plucked hairs, skin and nail scraping were examined and identified by direct microscopy and culture. Results From 4871 patients, 4015 strains were identified including dermatophytes (2635 cases, 65.7%), yeasts (1210 cases, 30.1%) and molds (170 cases, 4.2%). Among the dermatophytes, the most common pathogen isolated was Trichophyton rubrum (28.7%), followed by Epidermophyton floccosum (25.0%), Trichophyton mentagrophytes (23.8%) and Trichophyton tonsurans (11.5%). Among the yeast-like fungi, a predominance of Candida spp. (54.5%) was observed. Of these, 29.3% were C. albicans . Aspergillus spp. was the most prevalent isolated mold (71.8%). Conclusion Our results have demonstrated epidemiologic trends of dermatomycoses and their causative agents in recent years in comparison with the past records in Tehran and other parts of the world. This noticeable information can help to develop fungal diagnosis as well as more effective therapeutic methods.

In the present study, genetic diversity and mycotoxin profiles of Aspergillus flavus isolated from air (indoors and outdoors), levels (surfaces), and soils of five hospitals in Southwest Iran were examined. From a total of 146 Aspergillus colonies, 63 isolates were finally identified as A. flavus by a combination of colony morphology, microscopic criteria, and mycotoxin profiles. No Aspergillus parasiticus was isolated from examined samples. Chromatographic analyses of A. flavus isolates cultured on yeast extract-sucrose broth by tip culture method showed that approximately 10% and 45% of the isolates were able to produce aflatoxin B(1) (AFB(1)) and cyclopiazonic acid (CPA), respectively. Around 40% of the isolates produced sclerotia on Czapek-Dox agar. The isolates were classified into four chemotypes based on the ability to produce AF and CPA that majority of them (55.5%) belonged to chemotype IV comprising non-mycotoxigenic isolates. Random amplified polymorphic DNA (RAPD) profiles generated by a combination of four selected primers were used to assess genetic relatedness of 16 selected toxigenic and non-toxigenic isolates. The resulting dendrogram demonstrated the formation of two separate clusters for the A. flavus comprised both mycotoxigenic and non-toxigenic isolates in a random distribution. The obtained results in this study showed that RAPD profiling is a promising and efficient tool to determine intra-specific genetic variation among A. flavus populations from hospital environments. A. flavus isolates, either toxigenic or non-toxigenic, should be considered as potential threats for hospitalized patients due to their obvious role in the etiology of nosocomial aspergillosis.

In a search for novel antifungals from natural sources, essential oils (EOs) and extracts of 49 medicinal plants were studied against an aflatoxin (AF)-producing Aspergillus parasiticus using a microbioassay technique. AF levels were measured in culture broth by high performance liquid chromatography. The EOs were analyzed by gas chromatography/mass spectrometry (GC/MS). Based on the results obtained, Achillea millefolium subsp. elborsensis, Ferula gummosa, Mentha spicata, Heracleum pubescens and Thymus fedtschenkoi markedly inhibited A. parasiticus growth by IC50 values of 35 to 1,815 μg/ml without affecting AF production by the fungus. The EOs of flowers and roots of Foeniculum vulgare significantly inhibited both fungal growth (∼70.0%) and production of AFs B1 and G1 (∼99.0%). The ethyl acetate extract of Platycladus orientalis leaves suppressed AFB1 (∼90.0%) but not fungal growth and AFG1 production. This work provides evidence for the first time that F. vulgare and P. orientalis are strong inhibitors of aflatoxin biosynthesis in A. parasiticus. The antifungal activities of the bioactive plants introduced in the present study could make an important contribution to explaining the use of these plants as effective antimicrobial candidates to protect foods and feeds from toxigenic fungus growth and subsequent AF contamination.

The antifungal activity of Matricaria chamomilla L. flower essential oil was evaluated against Aspergillus niger with the emphasis on the plant's mode of action at the electron microscopy level. A total of 21 compounds were identified in the plant oil using gas chromatography/mass spectrometry (GC/MS) accounting for 92.86% of the oil composition. The main compounds identified were alpha-bisabolol (56.86%), trans-trans-farnesol (15.64%), cis-beta-farnesene (7.12%), guaiazulene (4.24%), alpha-cubebene (2.69%), alpha-bisabolol oxide A (2.19%) and chamazulene (2.18%). In the bioassay, A. niger was cultured on Potato Dextrose Broth medium in 6-well microplates in the presence of serial two fold concentrations of plant oil (15.62 to 1000 microg/mL) for 96 h at 28 degrees C. Based on the results obtained, A. niger growth was inhibited dose dependently with a maximum of approximately 92.50% at the highest oil concentration. A marked retardation in conidial production by the fungus was noticed in relation to the inhibition of hyphal growth. The main changes of hyphae observed by transmission electron microscopy were disruption of cytoplasmic membranes and intracellular organelles, detachment of plasma membrane from the cell wall, cytoplasm depletion, and complete disorganization of hyphal compartments. In scanning electron microscopy, swelling and deformation of hyphal tips, formation of short branches, and collapse of entire hyphae were the major changes observed. Morphological alterations might be due to the effect on cell permeability through direct interaction of M. chamomilla essential oil with the fungal plasma membrane. These findings indicate the potential of M. chamomilla L. essential oil in preventing fungal contamination and subsequent deterioration of stored food and other susceptible materials.

In order to find out plants useful to controlling aflatoxins (AFs) production, the essential oils (EOs) from 12 medicinal plants prepared by hydrodistillation were studied with special reference to the inhibition of Aspergillus parasiticus growth and AFs production. The toxigenic fungus was cultured in presence of various oils in 6-well microplates using a microbioassay technique. The mycelial mass was estimated as an index of fungal growth, while the aflatoxins B1 (AFB1) and G1 (AFG1) were measured by high performance liquid chromatography (HPLC). Among plants tested, Thymus vulgari and Citrus aurantifolia were found to inhibit both A. parasiticus and AF production. The EOs from Mentha spicata L., Foeniculum miller, Azadirachta indica A. Juss, Conium maculatum and Artemisia dracunculus were only inhibited fungal growth, while Carum carvi L. effectively inhibited AF production without any obvious effect on fungal growth. The other plants including Ferula gummosa, Citrus sinensis, Mentha longifolia and Eucalyptus camaldulensis had no effect on A. parasiticus growth and AF production at all concentrations used. The IC50 values of T. vulgaris, C. aurantifolia and C. carvi for AF inhibition were reported as 93.5, 285.6, and 621.9 μg/ml for AFB1, while they were calculated as 11.7, 50.1, and 56.0 μg/ml for AFG1. These results indicate that the EOs of some medicinal plants may be considered as potential candidates to protect foods and feeds from toxigenic fungus growth and subsequent AF contamination.

In an effort to screen the essential oils of some Iranian medicinal plants for novel aflatoxin (AF) inhibitors, Satureja hortensis L. was found as a potent inhibitor of aflatoxins B1 (AFB1) and G1(AFG1) production by Aspergillus parasiticus NRRL 2999. Fungal growth was also inhibited in a dose-dependent manner. Separation of the plant inhibitory substance(s) was achieved using initial fractionation of its effective part (leaf essential oil; LEO) by silica gel column chromatography and further separation by reverse phase-high performance liquid chromatography (RP-HPLC). These substances were finally identified as carvacrol and thymol, based on the interpretation of 1H and 13C NMR spectra. Microbioassay (MBA) on cell culture microplates contained potato-dextrose broth (PDB) medium (4 days at 28 degrees C) and subsequent analysis of cultures with HPLC technique revealed that both carvacrol and thymol were able to effectively inhibit fungal growth, AFB1 and AFG1 production in a dose-dependent manner at all two-fold concentrations from 0.041 to 1.32 mM. The IC50 values for growth inhibition were calculated as 0.79 and 0.86 mM for carvacrol and thymol, while for AFB1 and AFG1, it was reported as 0.50 and 0.06 mM for carvacrol and 0.69 and 0.55 mM for thymol. The results obtained in this study clearly show a new biological activity for S. hortensis L. as strong inhibition of aflatoxin production by A. parasiticus. Carvacrol and thymol, the effective constituents of S. hortensis L., may be useful to control aflatoxin contamination of susceptible crops in the field.

In the present study, diversity and phylogenetic relationship of Aspergillus species isolated from Tehran air was studied using random amplified polymorphic DNA (RAPD)–polymerase chain reaction (RAPD-PCR). Thirty-eight Aspergillus isolates belonging to 12 species i.e. A. niger (28.94 %, 11 isolates), A. flavus (18.42 %, 7 isolates), A. tubingensis (13.15 %, 5 isolates), A. japonicus (10.52 %, 4 isolates), A. ochraceus (10.52 %, 4 isolates), and 2.63 %, 1 isolate from each A. nidulans, A. amstelodami, A. oryzae, A. terreus, A. versicolor, A. flavipes and A. fumigatus were obtained by settle plate method which they were distributed in 18 out of 22 sampling sites examined. Fungal DNA was extracted from cultured mycelia of all Aspergillus isolates on Sabouraud Dextrose Agar and used for amplification of gene fragments in RAPD-PCR using 11 primers. RAPD-PCR data was analyzed using UPGMA software. Resulting dendrogram of combined selected primers including PM1, OPW-04, OPW-05, P160, P54, P10 and OPA14 indicated the distribution of 12 Aspergillus species in 8 major clusters. The similarity coefficient of all 38 Aspergillus isolates ranged from 0.02 to 0.40 indicating a wide degree of similarities and differences within and between species. Taken together, our results showed that various Aspergillus species including some important human pathogenic ones exist in the outdoor air of Tehran by different extents in distribution and diversity and suggested inter- and intra-species genetic diversity among Aspergillus species by RAPD-PCR as a rapid, sensitive and reproducible method.

Aflatoxin contamination of grains and agro-products is a serious food safety issue and a significant economic concern worldwide. In the present study, the effects of eugenol on Aspergillus parasiticus growth and aflatoxin production were studied in relation to the expression of some essential genes involved in aflatoxin biosynthetic pathway. The fungus was cultured in presence of serial two-fold concentrations of eugenol (15.62-500 μg mL(-1)) for 3 days at 28 °C. Mycelia dry weight was determined as an index of fungal growth, while aflatoxin production was assessed by high performance liquid chromatography. The expression of aflatoxin biosynthetic genes including ver-1, nor-1, pksA, omtA and aflR were evaluated by real-time PCR. Eugenol strongly inhibited A. parasiticus growth in the range of 19.16-95.83 % in a dose-dependent manner. Aflatoxin B1 production was also inhibited by the compound in the range of 15.07-98.0 %. The expressions of ver-1, nor-1, pksA, omtA and aflR genes were significantly suppressed by eugenol at concentrations of 62.5 and 125 μg mL(-1). These results indicate that eugenol may be considered as a good candidate to control toxigenic fungal growth and the subsequent contamination of food, feed and agricultural commodities by carcinogenic aflatoxins.

Virulence of 11 native strains of entomopathogenic fungi; Metarhizium anisopliae (three strains), Beauveria bassiana (six strains) and Lecanicillium psalliotae (two strains) collected from different parts of Iran, were studied against different developmental stages of Rhipicephalus (Boophilus) annulatus. After the exposure of ticks to the fungal strains in different concentrations (i.e. 10(3), 10(5), 10(7) conidia/ml), various parameters such as mortality rate and reproductive efficiency of engorged females, mortality of unfed tick larvae and eclosion percentage of infected eggs were evaluated to determine the fungal virulence. Based on the obtained results, five strains including M. anisopliae (IRAN 437 C and DEMI 001), B. bassiana (IRAN 403 C) and L. psalliotae (IRAN 468 C and IRAN 518 C) were found to be virulent to various stages of tick developmental cycle. Mortality rate of engorged females was found to be dose-dependent with regard to the conidial concentration used. Total mortality rates of 90-100%, 70% and 56.6% were observed for M. anisopliae (IRAN 437 C and DEMI 001), B. bassiana (IRAN 403 C) and L. psalliotae (IRAN 468 C), 6-11 days post inoculation (PI) with 10(7) conidia/ml, respectively. Most strains were able to inhibit egg laying by females in the range of 0-26% in different conidial concentrations. The results indicated that the mean egg laying of treated engorged tick females exposed to M. anisopliae (IRAN 437 C) was less than the mean values of those treated with other fungal strains. Results revealed 89.1%, 35.5% and 56.3% decrease in egg hatchability and 88.69%, 78.15% and 59.74% reduction in reproductive efficiency of the ticks using 10(7) conidia/ml of M. anisopliae (IRAN 437 C), B. bassiana (IRAN 403 C) and L. psalliotae (IRAN 468 C), respectively. In general, the entomopathogenic effects of native M. anisopliae and B. bassiana against various developmental stages of R. (B.) annulatus were confirmed in the present work. Likewise, although L. psalliotae, which was introduced for the first time as an entomopathogenic fungus against tick had not more than 13.3% mortality effect against adult females, but its effect on egg hatchability and reproductive efficiency was remarkable.

The effects of Akacid(plus), a novel member of guanidine-based polymeric compounds recently introduced as a potent inhibitor of fungal growth and aflatoxin biosynthesis were studied on Aspergillus parasiticus by transmission electron microscopy (TEM). The toxigenic fungus was cultured on yeast extract-sucrose broth in presence of serial two-fold concentrations of Akacid(plus) (1.5-96 microL/50 mL medium) for 96 h at 28 degrees C with shaking. Mycelial samples exposed to fungistatic concentrations of compound (1.5-48 microL) were processed for TEM. Corresponding to the growth inhibition, TEM observations revealed morphological anomalies in fungal compartments. The results demonstrated that Akacid(plus) targets the plasma membrane of the hyphae by its breaking down at variable intervals with the formation of small membrane-bound vesicles inside the cytoplasm, while no obvious damage was observed on the cell wall. A marked depletion of cytoplasmic contents of hyphae accompanied with lysis and disruption of membranes of major organelles such as nuclei, mitochondria and endoplasmic reticulum indicates that in high fungistatic concentrations, Akacid(plus) passes not only through the cell wall but also through the plasma membrane and then interact with membranous structures of the cytoplasmic organelles. Ultrastructural changes of fungal compartments exposed to Akacid(plus) in relation to the fungal growth and aflatoxin biosynthesis are discussed.

The effects of Akacid plus, a novel guanidine-based polymer first introduced as a biocidal and disinfectant agent were studied on Aspergillus parasiticus growth and its aflatoxin (AF) productivity. The fungus was cultured on yeast extract-sucrose (YES) broth in presence of various twofold serial dilutions of 25% Akacid plus (1.5-96 microL/50 mL medium) and then incubated in shaking condition with 150 rev./min at 28 degrees C for 96 h. Based on obtained results, Akacid plus was found to significantly inhibit both growth and aflatoxin B1 (AFB1) synthesis in very low concentrations in a dose-dependent manner. Fungal growth inhibition was determined in the range of 9.6-99.6% in mycelia exposed to the total concentration range of 1.5-48 microL. A final concentration of 96 microL was necessary to completely inhibit the growth of fungus. Under similar conditions, AFB1 synthesis was found to be strongly inhibited by 8.1-98.0% in presence of 1.5-24 microL Akacid plus with a maximum of 100% by 48 microL concentration. With respect to the unique physico-chemical properties of Akacid plus, its marked inhibitory effects on A. parasiticus growth and its AFB1 synthesis shown for the first time in this study make it a promising candidate for application in prevention programmes of AF contamination of susceptible crops.

International audience; In the present study, 193 Aspergillus strains were isolated from a total of 100 soil samples of pistachio orchards, which all of them were identified as Aspergillus flavus as the most abundant species of Aspergillus section Flavi existing in the environment. Approximately 59%, 81%, and 61% of the isolates were capable of producing aflatoxins (AFs), cyclopiazonic acid (CPA), and sclerotia, respectively. The isolates were classified into four chemotypes (I to IV) based on the ability to produce AFs and CPA. The resulting dendrogram of random amplified polymorphic DNA (RAPD) analysis of 24 selected A. flavus isolates demonstrated the formation of two separate clusters. Cluster 1 contained both aflatoxigenic and non-aflatoxigenic isolates (17 isolates), whereas cluster 2 comprised only aflatoxigenic isolates (7 isolates). All the isolates of cluster 2 produced significantly higher levels of AFs than those of cluster 1 and the isolates that produced both AFB(1) and AFB(2) were found only in cluster 2. RAPD genotyping allowed the differentiation of A. flavus from Aspergillus parasiticus as a closely related species within section Flavi. The present study has provided for the first time the relevant information on distribution and genetic diversity of different A. flavus populations from nontoxigenic to highly toxigenic enable to produce hazardous amounts of AFB(1) and CPA in soils of pistachio orchards. These fungi, either toxigenic or not-toxigenic, should be considered as potential threats for agriculture and public health.

The antifungal activity of onion (Allium cepa L.) on two important dermatophytes, Trichophyton rubrum and Trichophyton mentagrophytes, with special reference to morphological aspects was studied. Growth of both fungi was found to be strongly inhibited by aqueous onion extract (AOE) as a dose-dependent manner. The extract showed fungicidal effect for both fungi at concentrations >3.12% (v/v). The fungus T. mentagrophytes was more affected by the onion as compared to T. rubrum at all concentrations used. Morphological effects of onion exposure were examined in correlation with fungal growth. Corresponding to the growth inhibition, light and electron microscopy observations revealed morphological anomalies in hyphal compartments. The results demonstrated that AOE targets the cell membrane of the fungi as breaking down of both inner and outer membranes with consequent extrution of materials into the surrounding medium. Cytoplasmic membranes and other membranous structures of organelles, such as nuclei and mitochondria, were also disrupted. In correlation to the fungal growth, morphological alterations occurred to a less content for T. rubrum compared with T. mentagrophytes. The hyphae of T. rubrum were found to be mainly affected by converting to resistant forms, i.e., chlamidospores as a consequence of phenotype switching response to AOE. Plasmolysis accompanied by an almost complete depletion and disorganization of cytoplasmic structures were found to be the final event which led to cell death. Ultrastructural evidences obtained from this study strongly support that morphological changes of T. rubrum and T. mentagrophytes caused by AOE are associated with its fungistatic and fungicidal activities. With respect to the morphological results and the preliminary data on fungal biochemistry, a mechanism of action by interacting of AOE with thiol (-SH) groups present in essential compartments of the fungal cells was postulated.

Aflatoxins are a group of polyketide mycotoxins that are produced mainly by members of the genus Aspergillus. Production of these toxic secondary metabolites is closely related to fungal development (Keller et al., 2005; Jamali et al., 2012). Contamination of food, feed and agricultural commodities by aflatoxins poses enormous economic and serious health concerns because these chemicals are highly carcinogenic and can directly influence the structure of DNA. The resulting genetic defects can lead to fetal misdevelopment and miscarriages; aflatoxins are also known to suppress immune systems (Razzaghi-Abyaneh et al., 2013). In a global context, aflatoxin contamination is a constant concern between the 35N and 35S latitude where developing countries are mainly situated. With expanding boundaries of developing countries, aflatoxin contamination has become a persistent problem to those emerging areas (Shams-Ghahfarokhi et al., 2013). The continuing threat by aflatoxin contamination of food, feed and agricultural commodities to the world population has made aflatoxin research one of the most exciting and rapidly developing study areas of microbial toxins. The present research topic includes six review articles, three mini reviews and four original research articles. Contributors highlight current global health issues arising from aflatoxins and aflatoxigenic fungi and cover important aspects of aflatoxin research including contamination of crops, epidemiology, molecular biology and management strategies. Special attention is given to fungus-plant host interactions, biodiversity and biocontrol, sexual recombination in aflatoxigenic aspergilli, potential biomarkers for aflatoxin exposure in humans and safe storage programs. Perrone et al. (2014) reported the expected risk of a shift in aflatoxin problems toward new territories particular in South East of Europe due to increasing average temperatures. Giving an overview on genetic diversity of A. flavus populations in Europe, the authors stressed the importance of selecting stable atoxigenic A. flavus strains as biocontrol agents. In the review of climate change on A. flavus growth and aflatoxin production, Medina et al. (2014) focused on the potential impact of key environmental factors, such as water activity (aw), temperature and atmospheric CO2, and their interactions on ecology, growth and aflatoxin production by the A. flavus both in vitro and on maize. The authors showed that while such interacting abiotic factors have little effect on fungal growth, they however have a significant impact on aflatoxin biosynthetic gene expression and can stimulate the production of aflatoxins. In the insightful mini-review on sexual recombination in aflatoxin-producing Aspergillus species, Moore (2014) concisely summarized the potential negative impact of sexual recombination on the feasibility of using biological controls to reduce aflatoxin contamination of field crops. The author discussed specifically the implication of sexual recombination on the fate of two commercially available biopesticides: AF36 and Afla-Guard®. In the excellent review on the characteristics of A. flavus as well as the biocontrol strategy using non-toxigenic A. flavus strains, Ehrlich (2014) described the current state and outlook of this application in agricultural field. The author concluded that understanding genetic variations among A. flavus strains is critical for developing a robust biocontrol strategy, and it is unlikely that a “one size fits all” strategy will work for preharvest aflatoxin reduction. Host resistance is a very attractive area on aflatoxin research, and various aspects of A. flavus-plant host interaction were investigated with special focuses on mechanisms resistant to fungal growth and aflatoxin production (Dolezal et al., 2014; Fountain et al., 2014; Hruska et al., 2014; Scarpari et al., 2014; Shan and Williams, 2014). In the up-to-date review on environmental influences on aflatoxin production on maize, Fountain et al. (2014) detailed the history of research on this complex interaction and pointed out future directions for elucidating host resistance and susceptibility to A. flavus colonization in relation to abiotic stress such as drought and heat stresses, and oxidative stress in which aflatoxin may function as an antioxidant to the producing fungus. Utilizing an aflatoxigenic Green Fluorescence Protein (GFP) A. flavus strain, Hruska et al. (2014) investigated invasion, colonization and competition in maize kernels by this engineered strain. The authors showed that a decrease in aflatoxin production is correlated with depression of the aflatoxigenic population by the biocontrol strain, AF36, supporting the theory of competitive exclusion. Using a lipidomic approach to investigate A. flavus-maize interactions, Scarpari et al. (2014) suggested that A. flavus elicits the production of oxylipins in host plants, which function as signals for regulating aflatoxins biosynthesis, conidiogenesis and sclerotia formation. Their results highlighted the important role of maize oxylipins in driving secondary metabolism in A. flavus. In the microarray study to identify maize genes expressed during pathogen infection, Dolezal et al. (2014) found that metabolic processes are linked to defense responses, which include physical changes within the kernel as well as a disruption in kernel development. Shan and Williams (2014) provided a concise but clear overview of current knowledge about quantitative trait loci of corn related to aflatoxin contamination and ongoing efforts in the development of resistant corn lines. The authors concluded that the “phenotypic traits/data” established based on transcriptomics and proteomics approaches could be translated into the practices for improving corn resistance. Human exposure to aflatoxins is another challenging but not well studied area. In the concise overview of aflatoxin contamination of foods and related biomarker research, Mohd-Redzwan et al. (2013) described the historical problems related to aflatoxins in Malaysia, and how these problems have influenced the Malaysian population by highlighting the aflatoxin concentrations in basic food products and their comparison with established aflatoxin limits. The authors emphasized the importance of the legislation of law for a more controlled food production, legal enforcement to meet the set regulatory standards, and the improvement of pre and post-harvest techniques to reduce aflatoxin amounts in food and hence to decrease diseases in Malaysian population. In the comprehensive review about using microRNAs as specific molecular biomarkers in populations exposed to aflatoxins and as early markers for evidence of presence of or damage by hepatocellular carcinoma (HCC), Valencia-Quintana et al. (2014) described differential expression of microRNAs under specific conditions related not only to chemical and environmental pollutants but also to biological pollutants such as the presence of aflatoxins in humans and animals, and consequently, their influence over HCC. The authors provided findings that are important to toxicological research because microRNAs can be used to predict the toxicity of some compounds and will help to explore new treatments. In a well designed study aimed at enhancing antimycotic activities of known antifungal chemicals by natural compounds, Kim et al. (2014) successfully increased chemosensitization of a kresoxim methyl (Kre-Me), a natural fungicide from strobilurin class by chemically-synthesized benzo analogs. The authors found that among tested benzo analogs, octylgallate (OG) inhibits both growth and aflatoxin production by toxigenic aspergilli more efficiently. The study provided good evidence of remarkable synergism between OG and Kre-Me, which enhances the effectiveness of Kre-Me considerably. The efficient chemosensitizing capability of OG in increasing the efficacy of Kre-Me could reduce effective dosages of strobilurins and alleviate negative side effects associated with the current antifungal treatment. Finally, as one of the most important and practical issues on aflatoxin research, aflatoxin prevention and elimination, Villers (2014) presented laboratory and field data on an Ultra Hermetic™ storage system, which creates an unbreatheable atmosphere through insect and microorganism respiration alone, in preventing the exponential production and accumulation of aflatoxins. This system is proven useful during multi-month post-harvest safe storage tests of maize, rice and peanuts in hot, humid countries. The author further stressed the need for research on post-harvest protection against aflatoxin contamination by determining the frequency at which excessive aflatoxin levels are reached in the field vs. after months of post-harvest storage using this system. In conclusion, this research topic opens exciting perspectives on global health issues related to aflatoxins in the food chain and on the development of suitable strategies for preventing toxigenic fungal growth in field and storage, thereby reducing or eliminating subsequent aflatoxin contamination of our food supplies.

International audience; Soil isolates of Aspergillus section Flavi from Mazandaran and Semnan provinces with totally different climatic conditions in Iran were examined for aflatoxins (AFs; B and G types), cyclopiazonic acid (CPA) and sclerotia production. A total of 66 Aspergillus flavus group strains were identified from three species viz. Aspergillus flavus, Aspergillus parasiticus and Aspergillus nomius in both locations. A. flavus (87.9%) was found to be the prominent species followed by A. nomius (9.1%) and A. parasiticus (3.0%). Only 27.5% of A. flavus isolates were aflatoxigenic (B(1) or B(1) and B(2)), out of which approximately 75% were capable to producing CPA. All the A. parasiticus and A. nomius isolates produced AFs of both B (B(1) and B(2)) and G (G(1) and G(2)) types, but did not produce CPA. Sclerotia production was observed in only 4 isolates of A. flavus among all 66 isolates from three identified species. A. flavus isolates were classified into various chemotypes based on the ability to produce aflatoxins and CPA. In this study, a new naturally occurring toxigenic A. flavus chemotype comprising of two strains capable of producing more AFB(2) than AFB(1) has been identified. A relatively larger proportion of aflatoxigenic A. flavus strains were isolated from corn field soils of Mazandaran province which indicate a possible relationship between high levels of relative humidity and the incidence of aflatoxin-producing fungi. The importance of incidence of Aspergillus section Flavi in corn field soils regard to their mycotoxin production profiles and crop contamination with special reference to climatic conditions is discussed.

International audience; The expression of four aflatoxin (AF) biosynthetic pathway genes (aflD, aflO, aflP and aflQ) was evaluated in 24 Aspergillus flavus strains isolated from soils of pistachio orchards, with the aim of rapidly and accurately differentiating toxigenic from non-toxigenic strains. The amounts of AFB1 produced by 20 aflatoxigenic strains varied from 1.25 to 321.56 ng/mg fungal dry weights in YES medium. RT-PCR results showed that transcription of the four genes was not always correlated with AF production. The expression pattern of aflO and aflQ, however, was found to be well correlated with the amounts of AFB1 produced when strains were arbitrarily classified into two types: type I, comprised of strains producing ≥30 ng/mg; and type II, low (≤30 ng/mg) and non-AF producers. The present study suggests that, under specific growth conditions, the expression pattern of aflatoxin biosynthetic pathway genes such as aflO and aflQ can be used to infer the AF-producing capability of A. flavus strains.

Summary Objective The increasing importance of dermatophytoses and emerging resistance of dermatophytes to current synthetic antifungals have stimulated the search for safer and more effective alternative drugs from natural sources. The present study was carried out to identify antagonistic bacteria of soil origin with strong inhibitory activities on the growth of major human pathogenic dermatophytes. Materials and methods Antifungal activity of isolated soil bacteria was screened against the dermatophytes from three genera Microsporum (M. canis, M. gypseum), Epidermophyton (E. floccosum) and Trichophyton (T. mentagrophytes, T. rubrum, T. violaceum, T. tonsurans) by using visual plate agar assay method. A Pseudomonas chlororaphis isolate S105, identified at the species level by 16S ribosomal RNA sequence analysis, was reported as the strongest antagonistic bacterium. P. chlororaphis S105 culture supernatant (PCCS) was examined against tested dermatophytes by GY (glucose-yeast extract) broth bioassay in 6-well microplates. Antifungal compound of the bacterium was partially purified from the culture supernatant through a purification scheme of methanol extraction, Diaion HP20 ion-exchange chromatography and preparative thin layer chromatography. Results P. chlororaphis S105 was the most potent inhibitor of fungal growth for all tested dermatophytes with a percent inhibition ranged from 57.1% to 99.8%. The PCCS suppressed the growth of all fungi tested in the range of 18.5% to 84.8%. Partially purified antifungal compound of the bacterium was identified as a phenazine-like compound with an Rf value of 0.51. The compound inhibited fungal growth by 73.6% to 97.9% on GY broth. Fungal growth inhibition was significant for all dermatophytes tested in comparison with the controls (Anova, P Conclusion With respect to the strong inhibitory activity of P. chlororaphis against pathogenic dermatophytes reported here, it may be considered as a rich source of useful metabolites with potential application in antifungal drug discovery.