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MiRNAs are critical regulators of numerous physiological and pathological processes. Ascosphaera apis exclusively infects bee larvae and causes chalkbrood disease. However, the function and mechanism of miRNAs in the bee larval response to A. apis infection is poorly understood. Here, ame-miR-34, a previously predicted miRNA involved in the response of Apis mellifera larvae to A. apis invasion, was subjected to molecular validation, and overexpression and knockdown were then conducted to explore the regulatory functions of ame-miR-34 in larval body weight and immune response. Stem-loop RT-PCR and Sanger sequencing confirmed the authenticity of ame-miR-34 in the larval gut of A. mellifera. RT-qPCR results demonstrated that compared with that in the uninfected larval guts, the expression level of ame-miR-34 was significantly downregulated (p < 0.001) in the guts of A. apis-infected 4-, 5-, and 6-day-old larvae, indicative of the remarkable suppression of host ame-miR-34 due to A. apis infection. In comparison with the corresponding negative control (NC) groups, the expression level of ame-miR-34 in the larval guts in the mimic-miR-34 group was significantly upregulated (p < 0.001), while that in the inhibitor-miR-34 group was significantly downregulated (p < 0.01). Similarly, effective overexpression and knockdown of ame-miR-34 were achieved. In addition, the body weights of 5- and 6-day-old larvae were significantly increased compared with those in the mimic-NC group; the weights of 5-day-old larvae in the inhibitor-miR-34 group were significantly decreased in comparison with those in the inhibitor-NC group, while the weights of 4- and 6-day-old larvae in the inhibitor-miR-34 group were significantly increased, indicating the involvement of ame-miR-34 in modulating larval body weight. Furthermore, the expression levels of both hsp and abct in the guts of A. apis-infected 4-, 5-, and 6-day-old larvae were significantly upregulated after ame-miR-34 overexpression. In contrast, after ame-miR-34 knockdown, the expression levels of the aforementioned two key genes in the A. apis-infected 4-, 5-, and 6-day-old larval guts were significantly downregulated. Together, the results demonstrated that effective overexpression and knockdown of ame-miR-34 in both noninfected and A. apis-infected A. mellifera larval guts could be achieved by the feeding method, and ame-miR-34 exerted a regulatory function in the host immune response to A. apis invasion through positive regulation of the expression of hsp and abct. Our findings not only provide a valuable reference for the functional investigation of bee larval miRNAs but also reveal the regulatory role of ame-miR-34 in A. mellifera larval weight and immune response. Additionally, the results of this study may provide a promising molecular target for the treatment of chalkbrood disease.

Background: A high carriage rate of methicillin-resistant Staphylococcus aureus with the mecC gene (mecC-MRSA) has been described among Wild European hedgehogs (Europeaus erineaus). Due to this frequent occurrence, it has been suggested that hedgehogs could be a natural reservoir for mecC-MRSA. However, the reason why hedgehogs carry mecC-MRSA remains unknown, but it has been hypothesized that mecC-MRSA could have evolved on the skin of hedgehogs due to the co-occurrence with antibiotic producing dermatophytes. The aim of this pilot-study was therefore to investigate if hedgehogs in Sweden carry Trichophyton spp. and to provide evidence that these dermatophytes are able to produce penicillin or similar substances. In addition, the study aimed to identify if dermatophytes co-occurred with mecC-MRSA., Methods: Samples were collected from hedgehogs (Europeaus erineaus) that were euthanized or died of natural causes. All samples were screened for dermatophytes and mecC-MRSA using selective cultivation methods. Suspected isolates were characterized using PCR-based methods, genome sequencing and bioinformatic analyses. Identification of penicillin was performed by ultra-high-performance liquid chromatography-tandem mass spectrometry., Results: In total 23 hedgehogs were investigated, and it was shown that two carried Trichophyton erinacei producing benzyl-penicillin, and that these hedgehogs also carried mecC-MRSA. The study also showed that 60% of the hedgehogs carried mecC-MRSA., Conclusion: The pilot-study demonstrated that Trichophyton erinacei, isolated from Swedish hedgehogs, can produce benzylpenicillin and that these benzylpenicillin-producing T. erinacei co-occurred with mecC-MRSA. The study also reconfirmed the high occurrence of mecC-MRSA among hedgehogs., (© 2021. The Author(s).)

Dermatophytoses are inflammatory cutaneous mycoses caused by dermatophyte fungi of the genera Trichophyton, Microsporum, and Epidermophyton that affect both immunocompetent and immunocompromised individuals. With therapeutic failure, dermatophytoses can become chronic and recurrent. This is partly due to their ability to develop biofilms, microbial communities involved in a polymeric matrix attached to biotic or abiotic surfaces, contributing to fungal resistance. This review presents evidence accumulated in recent years on antidermatophyte biofilm activity. The following databases were used: Web of Science, Medline/PubMed (via the National Library of Medicine), Embase, and Scopus. Original articles published between 2011 and 2020, emphasizing the antifungal activity of conventional and new drugs against dermatophyte biofilms were eligible. A total of 11 articles met the inclusion criteria and were reviewed - the studies used in vitro and ex vivo (fragments of nails and hair) experimental models. The articles focused on reports of antibiofilm activity for conventional antifungals, natural drugs, and new therapeutic tools. The strains reported on were T. mentagrophytes, T. rubrum, T. tonsurans, M. canis, and M. gypseum. Between the studies, the wide variability of experimental conditions in vitro and ex vivo was observed. The data suggest the need for methodological standardization (at some minimum). This review systematically presents current studies involving agents that present antibiofilm activity against dermatophytes; and an overview of the ideal in vitro and ex vivo experimental conditions to guarantee biofilm formation that may assist future research., Lay Abstract: This review presents the current studies on the antibiofilm activities of drugs against dermatophytes and ideal experimental conditions, which might guarantee in vitro and ex vivo biofilm formation. It can be useful to examine the efficacy of new antimicrobial drugs against dermatophytes., (© The Author(s) 2021. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

Objective: The present study aimed to identify reference genes for qPCR analysis of T. rubrum growth in culture media which promote adhesion-inducing conditions to the host tissue., Methods: We investigated the suitability of six candidate reference genes: β-act, β-tub, ef1-α, gapdh, sdha and rpl2 in reference strain of Trichophyton rubrum in response to different environmental stimuli. The stability of these genes was determined by NormFinder, geNorm and BestKeeper software., Results: Our data obtained from the three algorithms revealed that mRNA expression levels of two candidate reference genes, ef1-α and β-tub, remained the most stable in response to different carbon sources, while different sample sets had their own most stable reference genes, highlighting the importance of the choice of internal controls in qPCR experiments. We then checked the stability of ef1-α and β-tub reference genes expression in different T. rubrum strains, suggesting that these two genes are reliable for normalisation of qPCR. Finally, we validated the suitability of selected reference genes as internal controls for target gene (SUB3) using the 2 -ΔΔCt method. The best result indicating an increase of SUB3 transcript of T. rubrum was found when the two the most stable reference (ef1-α and β-tub ) genes were used, as revealed by all three algorithms., Conclusions: We recommend the use of ef1-α and β-tub as reference genes for qPCR analysis of target gene expression in T. rubrum exposed to different carbon sources which promote adhesion-inducing conditions., (© 2020 Wiley-VCH GmbH.)

Chlamydoconidium-producing Trichophyton tonsurans strains isolated in Northeastern Brazil have morphological features different from the classic description of this dermatophyte species. This study investigated the phylogenetic relationship of chlamydoconidium-producing T. tonsurans strains isolated in Northeastern Brazil. Also, the effect of terbinafine and farnesol on mature biofilms of T. tonsurans strains was evaluated. The mass spectra of T. tonsurans strains were investigated by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The ITS and LSU loci regions of rDNA and the partial β-tubulin gene were sequenced and the phylogenetic tree was analysed. The effects of terbinafine and farnesol on mature T. tonsurans biofilms were evaluated through the analysis of metabolic activity, quantification of biomass and observation by scanning electron microscopy. MALDI-TOF MS spectra of the chlamydoconidium-producing T. tonsurans strains differed from the spectrum of the control strain (ATCC 28942), presenting an intense ion peak at m/z 4155 Da. Phylogenetic tree analysis showed that the chlamydoconidium-producing strains isolated in Northeastern Brazil are allocated to a single cluster, differing from strains isolated from other countries. As for mature T. tonsurans biofilms, farnesol reduced biomass and metabolic activity by 64.4 and 65.9 %, respectively, while terbinafine reduced the biomass by 66.5 % and the metabolic activity by 69 %. Atypical morphological characteristics presented by chlamydoconidium-producing T. tonsurans strains result from phenotypic plasticity, possibly for adaptation to environmental stressors. Also, farnesol had inhibitory activity against T. tonsurans biofilms, demonstrating this substance can be explored for development of promising anti-biofilm drugs against dermatophytes.

Aims: According to the WHO, 20-25% of people worldwide are affected by skin infections caused by dermatophytes, such as those of the Trichophyton genus. Additionally, several dermatophytes have developed resistance to drugs such as griseofulvin and itraconazole. This study tested 2S albumins-derived antimicrobial peptides (AMPs) as alternative antidermatophytic molecules., Main Methods: Membrane pore formation assays, tests to detect overproduction of ROS, scanning electron microscopy (SEM) and fluorescence microscopy (FM) were carried out to provide insight into the mechanisms of antidermatophytic action., Key Findings: All AMPs (at 50 μg mL -1 ) tested reduced the mycelial growth of T. mentagrophytes and T. rubrum by up to 95%. In contrast, using a concentration 20-fold higher, griseofulvin only inhibited T. mentagrophytes by 35%, while itraconazole was not active against both dermatophytes. Scanning electron and fluorescence microscopies revealed that the six AMPs caused severe damage to hyphal morphology by inducing cell wall rupture, hyphal content leakage, and death. Peptides also induced membrane pore formation and oxidative stress by overproduction of ROS. Based on the stronger activity of peptides than the commercial drugs and the mechanism of action, all six peptides have the potential to be either employed as models to develop new antidermatophytic drugs or as adjuvants to existing ones., Significance: The synthetic peptides are more efficient than conventional drug to treat infection caused by dermatophytes being potential molecules to develop new drugs., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Trichophyton rubrum (Tr) is the main aetiological agent of human dermatophytosis, being isolated from the environment and keratinised tissues. In the environment, Tr can interact with other organisms, such as free-living amoebas (FLA), which can act as an alternative host system to study the interaction between microbes and phagocytic cells., Objectives: To characterise the Acanthamoeba castellanii (ALX)-Tr interaction., Methods: Interaction was characterised in three conditions: trophozoites (PYG), late (PYG/NES) and early (NES) encystation stimulus, evaluating encystation kinetics, phagocytosis, exocytosis and fungicidal activity dynamics., Results: Tr was able to induce ALX encystation and be internalised by ALX. The number of internalised conidia was high at 1 hour, and ALX presented fungicidal activity with increased intracellular ROS production and exocytosis. In PYG/NES, phagocytosis and ROS production were reduced, with decreased ALX's fungicidal activity. However, in NES there was an increased fungal engulfment, and a reduced ROS production and higher fungal burden. Furthermore, exogenous mannose decreased phagocytosis of Tr conidia, and divalent cations induced ROS production and increased ALX's fungicidal activity. Interestingly, phagocytosis was reduced in the presence of cytoskeleton inhibitor, but exocytosis was increased, suggesting that Tr conidia may have alternative pathways to escape ALX's cells., Conclusion: A castellanii is a proper model for studying Tr-FLA interaction, since ALX can engulf, produce ROS and kill Tr, and all these parameters are influenced by an encystation stimulus and divalent cations. Moreover, this interaction is likely to occur in the environment implicating in the adaptation to environmental stressful conditions in both organisms., (© 2020 Wiley-VCH GmbH.)

The increasing number of dermatophytoses among animals observed recently in developed countries may be connected with relocation of many exotic species outside their natural living environment. Moreover, an impact on this situation may also be exerted by relapses related to limited compliance with antifungal treatment regimes. Many exotic animals, including camelids, imported to European countries are connected with cases of tuberculosis or zoophilic dermatophytoses in humans. In the present study, we identified and comprehensively analysed dermatophyte infections in alpacas from breeding farms in Poland. As part of this study, we determined the prevalence of dermatophyte infections in alpacas. The conventional and molecular mycological diagnostic procedures applied led to unambiguous identification of the aetiological factors of symptomatic dermatophytoses and asymptomatic animals, that is Trichophyton benhamiae and T. verrucosum. Furthermore, the susceptibility tests allowed choosing the best therapeutic option and revealed superior activity of allylamine drugs against all strains. Finally, in the case of strains isolated from symptomatic dermatophytoses, our study revealed a significantly higher virulence level expressed by high activity of chosen enzymes, especially related to keratinolytic and haemolytic activity. In conclusion, this report indicates that farmed alpacas can be a reservoir or vector for contagious zoophilic dermatophyte infection. For this reason, imported animals should be subjected to meticulous monitoring to detect not only symptomatic infections but also asymptomatic animals., (© 2020 Blackwell Verlag GmbH.)

Each year, millions of people worldwide are affected by superficial mycoses, which are frequently caused by dermatophytes having affinity to the scalp, nails, hair and the stratum corneum of the skin. The changing reservoirs of zoophilic dermatophytes, the numerous reports of animal carrier status, and the increasing number of pets make the diagnosis difficult, which is usually impossible based only on medical history. Herein we present a case report of tinea corporis caused by Trichophyton equinum in siblings who had no contact with horses. The routine laboratory diagnostic procedures for identification of isolates were based on phenotypic and genotypic characteristics, especially molecular techniques using rDNA internal transcribed spacer sequences. The results showed that both techniques proved to be insufficiently discriminatory to differentiate two closely related species, i.e. Trichophyton equinum and Trichophyton tonsurans. Introduction of a TEF1 sequence analysis to the diagnostic procedures revealed consistent differences between these two species and facilitated unambiguous identification. Interestingly, dogs that could leave the homestead freely were the source of the infection in children. In conclusion, Trichophyton equinum was considered in the past as a strict zoophilic dermatophyte associated with horses and rarely transmitted to humans. This study revealed that this species can have other reservoirs and live in the fur of asymptomatic animals.

Objective: In order to improve the effect of ketoconazole, poly-lactic acid (PLA) nanoparticles containing ketoconazole were prepared, characterized and tested against dermatophytes and Candida spp planktonic and biofilm cells., Methods: The ketoconazole-PLA nanoparticles obtained by nanoprecipitation were characterized using dynamic light scattering, scanning electron microscopy, transmission electron microscopy, and differential scanning calorimetry. In addition, quantification of encapsulated ketoconazole and the in vitro release profile were determined. Antifungal susceptibility tests against dermatophytes Trichophyton rubrum, Trichophyton mentagrophytes, and Microsporum gypseum and yeasts Candida albicans, C. dubliniensis, C. krusei, C. parapsilosis, and C. tropicalis were performed., Results: Spherical nanoparticles, with a mean diameter of 188.5nm and an encapsulation efficiency of 45% ketoconazole, were obtained. The nanoparticles containing ketoconazole had superior antifungal activity against all tested fungi strains than free ketoconazole. Inhibition of yeast biofilm formation was also achieved., Conclusion: Ketoconazole-PLA nanoparticles resulted in better antifungal activity of ketoconazole nanoparticles than free drug against dermatophytes and Candida species, indicating a promising tool for the development of therapeutic strategies., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Under steady-state conditions, the immune system is poised to sense and respond to the microbiota. As such, immunity to the microbiota, including T cell responses, is expected to precede any inflammatory trigger. How this pool of preformed microbiota-specific T cells contributes to tissue pathologies remains unclear. Here, using an experimental model of psoriasis, we show that recall responses to commensal skin fungi can significantly aggravate tissue inflammation. Enhanced pathology caused by fungi preexposure depends on Th17 responses and neutrophil extracellular traps and recapitulates features of the transcriptional landscape of human lesional psoriatic skin. Together, our results propose that recall responses directed to skin fungi can directly promote skin inflammation and that exploration of tissue inflammation should be assessed in the context of recall responses to the microbiota., Competing Interests: Competing interest statement: D.K. and S.N. are coauthors on a 2012 original article and a 2018 review paper. S.T. is an employee of L’Oréal Research and Innovation.

Difficulties in obtaining human nails that are large enough for examining the penetration of drug formulations led us to produce keratin films regenerated from human hair. We assume that these films can simulate human nail plates in drug penetration and permeation tests and can serve as a biological model for studying onychomycosis. The films were formed from keratin extracted from human hair using dithiothreitol, urea and thiourea. The obtained keratin extract was dispensed into Teflon rings and dried at 40 °C and then cured at 110 °C. The structure, surface morphology, chemical characterization and thermal stability of the films were characterized and were compared to those of human nail, hair and bovine hoof samples using SDS-electrophoresis, scanning electron microscopy (SEM), X-ray diffraction analysis (XRD), Fourier transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA). The structure of the obtained films was found to be closer to human nails than to hair or bovine hooves. The keratin films were infected with Trichophyton rubrum and were proven to be appropriate for serving as a model for studying onychomycosis.

The delineation of the pathogenic interaction between the host skin immune responses and dermatophytes has remained indigent. The obscure enigma in host-dermatophyte immunopathogenic interactions is the T regulatory (Treg) and T-helper (Th) 17  cell role in maintaining immune homeostasis. We attempted to understand the regulation and recognition of lineage-specific response in chronic dermatophytic skin infection patients. The percentages of Th17 (CD4 + CD161 + IL23R + ) and Treg (CD4 + CD25 + FoxP3 + ) cell subpopulations in the peripheral circulation of thirty chronic dermatophytic skin infection patients and twenty healthy individuals was determined. The serum cytokine levels were estimated for disease correlation. The mean duration of the disease was 10.68 ± 8.72 months, with Trichophyton mentagrophytes complex as the major pathogen. Total serum IgE level of patients was significantly higher compared to healthy controls (305 ± 117 vs 98.53 ± 54.55 IU/ml; p < 0.01). Expression of Th17 and Treg cell markers on CD4 + T cells was significantly elevated in patients than controls (p < 0.05). Comparatively, serum interleukin (IL)-4 and interferon (IFN)-γ levels were increased, with low IL-10 levels in patients. Our data envisages a complex immune dysfunction in chronic dermatophytosis, arising either as a result of dermatophyte exposure or paradoxical precedence of disease establishment. Designing new treatment strategies and preventing recurrences are challenges for future research., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Dermatophytes are a highly specialized group of keratinophilic and keratinolytic filamentous fungi causing a ringworm disease called dermatophytosis or superficial mycoses. Although dermatophyte infections do not threaten the host's life, they lower its quality in humans by causing discomfort related to cosmetic problems and through their epidemiological significance, whereas in farm animals they are responsible for economic losses and constitute a source of the spread of spores. Evidence from countless observational studies that have been conducted over the last 90 years indicates that dermatophytes infect humans of every age, race, gender and socioeconomic status with strikingly high rates, as well as both farmed and wild animals in various health conditions and with various epidemiological statuses. However, the prevalence of superficial fungal infections is highly variable, since it depends on several parameters associated with the infected individual and the dermatophyte, their mutual interactions, and epidemiological and geographical factors. The curious disparity in dermatophyte infection patterns has prompted many investigators to search for a link between the host, the host's predispositions and susceptibility to the disease, and the dermatophyte species and virulence. Thus, the question arises as to whether, in addition to the generally recognized factors predisposing hosts to diseases, there are some other predispositions to dermatophyte infections in a species-specific host. In this review, we describe recent findings about the mechanism of dermatophyte infections, focusing on the adaptation of the fungi to the host and conditions predisposing each side to the disease.

Dermatophyte infections are extremely frequent worldwide, and their epidemiological features and distribution make them one of the most frequent infections all over the world. We identified and analysed multiform T. mentagrophytes strains isolated from a silver fox (Vulpes vulpes) kept on a breeding farm. Identification of dermatophyte strains was carried out traditionally by correlating both the clinical manifestations of the infection with a micro- and macroscopic examination. To confirm the species affiliation fully, molecular differentiation methods were used. DNA was isolated from the dermatophytes with the phenol-chloroform method. The reaction of chitin synthase 1 (chs1) amplification was carried out to confirm the dermatophytes. The phylogenetic analysis was based on the ITS sequences. The polymerase chain reaction melting profile (PCR-MP) procedure was used for differentiation of dermatophyte genomes. Direct analysis of the material sampled from the clinical lesions revealed the presence of arthrospores in the samples collected from all animals with skin lesions. The macromorphology of the colonies obtained from material sampled from the same individual was not homogeneous. The PCR-MP electrophoregram indicated high variability of their genomes. Although the dermatophytes were isolated from one infected fox, no two identical genomic profiles were obtained. The PCR-MP result corresponds with the phenotypic diversity of the isolates. The findings about the multiple dermatophyte infection in one individual complicate any future epidemiology work and other clinical investigation. Previously, using only morphological characteristics, it had been assumed that one fungal isolate per patient could be diagnosed. The novel findings encourage application of the newly developed molecular typing methods in the diagnosis of dermatophytosis.

A 68-year-old woman was submitted to our hospital because of erythematous and scaly skin lesions. To exclude tinea samples of stratum corneum were collected and used for mycological investigations. In this material, no fungal elements were detected microscopically, but inoculation on Sabouraud agar with cycloheximide yielded a presumptive dermatophyte fungus. Subsequent detailed investigations with conventional morphological and physiological methods and a phylogenetic analysis of the combined LSU rRNA gene (D1/D2 domains) and ITS region sequences suggested that the fungus represents a hitherto undescribed species of the genus Arthroderma. Here, we describe this species as Arthroderma chiloniense sp. nov., EMBL accession no. LT992885. This new species can be distinguished from phylogenetically related Arthroderma species using ribosomal ITS and LSU genes, and 60S L10 protein sequences; specific macroscopic, microscopic and physiological features are lacking. Our attempts to re-isolate this fungus from the patient's skin failed although her skin lesions persisted. Most likely A. chiloniense is a geophilic species that incidentally contaminated or transiently colonised the patient's skin. To avoid diagnostic misinterpretations, it is necessary to distinguish A. chiloniense from truly pathogenic dermatophytes like Trichophyton (T.) rubrum and T. interdigitale which can easily be confused with A. chiloniense based on similar mycelium morphology., (© 2018 Blackwell Verlag GmbH.)

Objectives: The aim of this study was to evaluate the antifungal, antichemotactic and antioxidant activities of Schinus lentiscifolius essential oil, as well as its combined effect with terbinafine and ciclopirox, against dermatophytes., Methods: Essential oil was analysed by GC-MS. The antifungal activity and the mechanism of action were determined by broth microdilution, sorbitol and ergosterol assays, as well as scanning electron microscopy. The checkerboard method was used for evaluating the interactions with commercial antifungal agents. The antioxidant and antichemotactic activities were measured using the DPPH and the modified Boyden chamber methods, respectively., Key Findings: Chemical analysis revealed the presence of 33 compounds, the primary ones being γ-eudesmol (12.8%) and elemol (10.5%). The oil exhibited 97.4% of antichemotactic activity and 37.9% of antioxidant activity. Antifungal screening showed effect against dermatophytes with minimum inhibitory concentration values of 125 and 250 μg/ml. Regarding the mechanisms of action, the assays showed that the oil can act on the fungal cell wall and membrane. Synergistic interactions were observed using the combination with antifungals, primarily terbinafine., Conclusions: Schinus lentiscifolius essential oil acted as a chemosensitizer of the fungal cell to the drug, resulting in an improvement in the antifungal effect. Therefore, this combination can be considered as an alternative for the topical treatment of dermatophytosis., (© 2018 Royal Pharmaceutical Society.)

Objectives: This study was conducted to evaluate the antidermatophytic activity of 48 extracts obtained from medicinal plants (Cibotium barometz, Melastoma malabathricum, Meuhlenbeckia platyclada, Rhapis excelsa, Syzygium myrtifolium, Vernonia amygdalina) and marine algae (Caulerpa sertularioides, Kappaphycus alvarezii) against Trichophyton rubrum and Trichophyton interdigitale (ATCC reference strains), and the cytotoxicity using African monkey kidney epithelial (Vero) cells. Active plant extracts were screened for the presence of phytochemicals and tested against clinical isolates of Trichophyton tonsurans., Methods: Six different extracts (hexane, chloroform, ethyl acetate, ethanol, methanol and water) were obtained from each plant or algae sample using sequential solvent extraction. The antidermatophytic activity for the extracts was assessed using a colourimetric broth microdilution method. The viability of Vero cells was measured by Neutral Red uptake assay., Results: All the extracts (except the water extracts of V. amygdalina, C. sertularioides and K. alvarezii) showed antidermatophytic activity against Trichophyton spp. The minimum fungicidal concentration (MFC) ranges for the plant extracts against T. rubrum and T. interdigitale are 0.0025-2.50 and 0.005-2.50mg/mL, respectively. The algae extracts exhibited lower potency against both species, showing MFC ranges of 0.08-2.50 and 0.31-2.50mg/mL, respectively. The ethanol and methanol extracts from the leaves of R. excelsa, and the methanol and water extracts from the leaves of S. myrtifolium were highly active (MFC<0.1mg/mL) and with high selectivity indices (SI>2.8) against reference strains of T. rubrum and T. interdigitale, and most of the clinical isolates of T. tonsurans. Phytochemical analysis indicates the presence of alkaloids, anthraquinones, flavonoids, saponins, tannins, phenolics and triterpenoids in the extracts., Conclusions: The medicinal plant extracts exhibited stronger antidermatophytic activity compared to the algae extracts. The leaves of R. excelsa and S. myrtifolium are potential sources of new antidermatophytic agents against Trichophyton spp., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

We report 2 new cases of invasive infections caused by Nannizziopsis spp. molds in France. Both patients had cerebral abscesses and were immunocompromised. Both patients had recently spent time in Africa.

Fungal diseases of the nail bed (onychomycosis) and epidermis are recurrent illnesses in the elderly and immunocompromised patients, which have few efficacious treatment options. Current treatment options for onychomycosis are limited to topical agents, laser treatment, and oral antifungals. Previous generations of topical agents were not efficacious, owing to poor penetration of the nail bed. Oral antifungal drugs, such as itraconazole, terbinafine, and fluconazole, not only give better response rates but also inhibit a host of CYP450 enzymes. Oral antifungals can exacerbate drug-drug interactions for patients taking other medications concurrently. Newer topical agents might recognize improved efficacy and provide therapeutic alternatives when the use of oral antifungal agents is contraindicated. Recently, the Food and Drug Administration (FDA) approved efinaconazole and tavaborole for the treatment of onychomycosis. Additionally, the FDA approved luliconazole for the treatment of tinea pedis, tinea cruris, and tinea corporis. This review examines the mechanism of action, spectrum of activity, pharmacokinetics, and clinical trials data and considers the place in therapy for these 3 new antimycotic agents.

Tinea unguium caused by dermatophyte species are usually treated with oral antimycotic, terbinafine (TBF). To understand the mechanisms of improvement and recalcitrance of tinea unguium by oral TBF treatment, a method of quantifying dermatophyte viability in the nail was developed, and the viability of dermatophytes was analyzed in toenail lesions of 14 patients with KOH-positive tinea unguium treated with oral TBF 125 mg/day for up to 16 weeks. Mycological tests, including KOH examination and fungal culture, and targeted quantitative real-time PCR for internal transcribed spacer (ITS) region, including rRNA, were demonstrated at the initial visit and after 8 and 16 weeks of treatment. Assays in eight patients showed that average ITS DNA amount significantly decreased, to 44% at 8 weeks and 36% at 16 weeks compared with 100% at initial visit. No significant difference was observed between at 8 and 16 weeks, despite the TBF concentration in the nail supposedly more than 10-fold higher than the minimum fungicidal concentration for dermatophytes. This finding suggests the pathogenic dermatophytes in nail lesions could survive in a dormant form, such as arthroconidia, during oral TBF treatment. Both antimycotic activity and nail growth are important factors in treatment of tinea unguium.

Dermatophytes are a group of closely related fungi that nourish on keratinized materials for their survival. They infect stratum corneum, nails, and hair of human and animals, accounting the largest portion of fungi causing superficial mycoses. Huge populations are suffering from dermatophytoses, though the biology of these fungi is largely unknown yet. Reasons are partially attributed to the poor amenability of dermatophytes to genetic manipulation. However, advancements in this field over the last decade made it possible to conduct genetic studies to satisfying extents. These included genetic transformation methods, indispensable molecular tools, i.e., dominant selectable markers, inducible promoter, and marker recycling system, along with improving homologous recombination frequency and gene silencing. Furthermore, annotated genome sequences of several dermatophytic species have recently been available, ensuring an optimal recruitment of the molecular tools to expand our knowledge on these fungi. In conclusion, the establishment of basic molecular tools and the availability of genomic data will open a new era that might change our understanding on the biology and pathogenicity of this fungal group.

Most superficial fungal infections are caused by dermatophytes, a specialized group of filamentous fungi which exclusively infect keratinized host structures such as hair, skin and nails. Since little is known about the molecular basis of pathogenicity and sexual reproduction in dermatophytes, here we functionally addressed two central transcriptional regulators, SteA and StuA. In the zoophilic species Arthroderma benhamiae a strategy for targeted genetic manipulation was recently established, and moreover, the species is teleomorphic and thus allows performing assays based on mating. By comparative genome analysis homologs of the developmental regulators SteA and StuA were identified in A. benhamiae. Knock-out mutants of the corresponding genes as well as complemented strains were generated and phenotypically characterized. In contrast to A. benhamiae wild type and complemented strains, both mutants failed to produce sexual reproductive structures in mating experiments. Analysis of growth on keratin substrates indicated that loss of steA resulted in the inability of ΔsteA mutants to produce hair perforation organs, but did not affect mycelia formation during growth on hair and nails. By contrast, ΔstuA mutants displayed a severe growth defect on these substrates, but were still able to produce hair perforations. Hence, formation of hair perforation organs and fungal growth on hair per se are differentially regulated processes. Our findings on the major role of SteA and StuA during sexual development and keratin degradation in A. benhamiae provide insights into their role in dermatophytes and further enhance our knowledge of basic biology and pathogenicity of these fungi.

Onychomycosis is a chronic fungal nail infection caused by dermatophytes, yeasts or non-dermatophyte moulds (NDMs). It occurs worldwide and accounts for about 50% of all nail disorders. The incidence of the disease and the prevalence of pathogens involved vary in different geographical areas and change with time. This study was undertaken to determine the epidemiology of onychomycosis in Crete, Greece during a 12-year period. Mycologic nail investigation (direct microscopy and cultures) was performed in samples taken from 3317 patients with clinical suspicion of onychomycosis. Demographic characteristics, comorbidities and concurrent superficial fungal infections at other body sites were also recorded. Onychomycosis was mycologically confirmed in 547 (16.5%) patients. Fingernail onychomycosis was found in 274 (50.1%) patients, toenail onychomycosis in 271 (49.5%), and both toenail and fingernail onychomycosis in two (0.4%). Yeasts were the major causative agents, followed by dermatophytes and NDMs. Candida albicans was the most commonly isolated yeast species and Trichophyton rubrum the most common dermatophyte. Concomitant fungal skin infections were noted in 53 (19.6%) patients with toenail onychomycosis. Factors predisposing to onychomycosis were trauma, diabetes, psoriasis, peripheral circulatory insufficiency and immunosuppression. Mycological investigations are necessary to confirm clinical diagnosis and identify the pathogen, in order to select the most appropriate treatment., (© 2016 Blackwell Verlag GmbH.)

Conventional treatments for onychomycosis include oral and topical antifungal agents. Although Nd:YAG laser has been reported to be effective, controlled studies combined or compared with conventional treatments are needed. To evaluate the efficacy of 1064 nm Nd:YAG laser for onychomycosis and compare outcomes with those of topical antifungal treatment and combination therapy. Patients were randomly divided into three groups: Laser (L); laser with topical antifungal therapy (L + T); and topical antifungal treatment (T). Laser treatment consisted of three or four sessions at 4 week intervals. Outcomes were assessed clinically and mycologically. In 217 nails of 56 patients, 76% in the L group and 71.8% in the L + T group experienced clinical responses, and 15.2% and 22.5%, respectively, were cured at 24 weeks. The clinical and cure rates of both groups were significantly greater than those of the T group. Six patients of the L group developed new infections, as did one patient in the L + T group and two patients in the T group. 1064 nm Nd:YAG laser shows clinically good responses in onychomycosis. Addition of topical antifungal agent did not appear to improve efficacy, but may have protected against new infection., (© 2016 Blackwell Verlag GmbH.)

Non-dermatophytic moulds (NDMs) have been increasingly recognised as causative agents of onychomycosis. The diagnosis of onychomycosis is most often obtained by microscopic observation of nail specimens where fungal elements can be detected and cultured by standard mycological techniques. Direct microscopic examination does not always result positive in NDM onychomycosis; therefore to perform a correct diagnosis, a proper mycological culture is often required. The purpose of our study was to evaluate the role of direct microscopic examination in the NDM onychomycosis diagnosis. The results show that only 57.2% of the specimens from onychomycosis patients could be properly diagnosed showing positivity to both direct microscopic examination and NDMs culture isolation in two or more subsequent inoculations, while 42.8% of analysed specimens with a negative direct microscopic examination, showed NDMs growth after three or more subsequent inoculations. The large proportion of false negatives (more than 42%) could be related to the duration of the infection and/or to the experience and skills of the personnel dedicated to specimen collection. We point out the need for thoroughly evaluating all specimens showing cultural growth in at least three subsequent medium inoculations, whatever the result of the microscopic examination, in order to reduce false-negative rates. This strategy would allow for more accurate diagnosis of this mycosis., (© 2016 Blackwell Verlag GmbH.)

Background: Anamorphs of Arthroderma (A.) benhamiae, which can cause inflammatory tinea lesions in humans, have been progressively spreading in Germany., Objectives: Identification of A. benhamiae anamorphs by conventional methods., Materials and Methods: Evaluation of our own results obtained with A. benhamiae anamorphs and from the relevant literature., Results: Infections with A. benhamiae anamorphs are usually transferred by guinea pigs or other animals. A. benhamiae anamorphs can be cultured on growth media used for dermatophytes and can be characterized morphologically and physiologically. In Germany the yellow variant is seen most often but a white variant that equals Trichophyton (T.) interdigitale has also been observed. On Sabouraud agar the yellow variant develops markedly yellow thalli, whereas the white variant produces white aerial mycelium. Microconidia are formed by the yellow variant-if at all-only scarcely and delayed; they are small, roundish, and arranged in a grape-like order. The white variant produces peg-shaped microconidia alongside hyphae as well as roundish ones in grape-like clusters, and subsequently some macroconidia, chlamydospores, and spiral hyphae. In microcultures with both variants circuit-like hyphal structures can consistently be demonstrated. On Trichophyton agars only the yellow variant is clearly dependent on thiamine. The urease test is negative with the yellow variant and positive with the white variant. Most strains of both variants are negative in the hair perforation test., Conclusions: The characteristics described allow reliable identification of the yellow variant of the A. benhamiae anamorph by conventional methods; a distinction between the white variant and T. interdigitale can be more difficult. Dermatologists should be able to identify this agent in clinical routine.

Dermatophytes are classified in three genera, Epidermophyton, Microsporum and Trichophyton. They have the capacity to invade keratinized tissue to produce a cutaneous infection known as dermatophytoses. This investigation was performed to study the effect of gaseous ozone and ozonized oil on three specific properties of six different dermatophytes. These properties included sporulation, mycelia leakage of sugar and nutrients and the activity of their hydrolytic enzymes. Generally, ozonized oil was found to be more efficacious than gaseous ozone. Microsporum gypseum and Microsporum canis were the most susceptible, while Trichophyton interdigitale and T. mentagrophytes were relatively resistant. The study revealed a steady decline in spore production of M. gypseum and M. canis on application of ozonated oil. An increase in leakage of electrolytes and sugar was noticed after treatment with ozonized oil in the case of M. gypseum, M. canis, T. interdigitale, T. mentagrophytes and T. rubrum. The results also revealed loss in urease, amylase, alkaline phosphatase, lipase and keratinase enzyme producing capacity of the investigated fungi., (Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.)

Dermatophytosis, which is caused mainly by genera of Trichophyton, Epidermophyton, and Microsporum, is a frequent dermatological problem in tropical and subtropical countries. Investigations were carried out in this study to evaluate the antidermatophytic activity of the stems, leaves, and seeds of Croton tiglium, one of the traditional medicine plants indigenous to Asia. Ethanolic extracts of the stems, leaves, and seeds of C. tiglium were prepared by cold soak or heat reflux methods. The antidermatophytic activities of the extracts were evaluated by disc diffusion and microdilution susceptibility assays against Trichophyton mentagrophytes, T. rubrum, and Epidermophyton floccosum. The active components in the extracts were analyzed and identified by GC-MS. All ethanolic extracts of C. tiglium showed some antifungal activities against the three dermatophytes. The ethanolic stem extract had the greatest inhibitory activities against T. mentagrophytes and E. floccosum with MICs at 0.16 mg/mL and had a lower activity against T. rubrum (MIC: 0.31 mg/mL). Oleic acid and hexadecanoic acid were found to be the major constituents in the stem extract that demonstrated strong antidermatophytic activities. The ethanolic extracts of stem or seed of C. tiglium exhibit strong antidermatophytic activities and, thus, could be considered for application on treating skin fungal infections after appropriate processing.

In the last decades, the number of people suffering from dermatophytoses has seriously increased, which may be due to the development of resistant strains to a range of antifungal drugs. The present study was aimed to evaluate the antidermatophytic properties of eight extracts from the selected spices and herbs, which were ethno-medicinally used in Iran against Trichophyton mentagrophytes, Trichophyton interdigitale, Microsporum canis, and Microsporum gypseum (10 strain of each). The in vitro antifungal activities of the extracts from four spices and four plants were evaluated by the broth macro dilution method against four dermatophyte strains. In addition, the in vivo therapeutic effects of Myrtus communis L. and Cinnamomum zeylanicum Blume extracts (the most active extracts) on dermatophytosis induced by M. canis and T. mentagrophytes in guinea pigs were evaluated. Results of in vitro antifungal assay revealed that all the tested extracts demonstrated both fungistatic and fungicidal activities with the geometric mean (GM) MIC ranging from 0.058 to 3.73 mg/ml and GM (MFC) ranging from 0.058 to 7.46 mg/ml, respectively. Two extracts (M. communis and C. zeylanicum) significantly inhibited the growth of all the tested dermatophytes, while other extracts demonstrated weak (MICs of >0.625 mg/ml) to moderate (MICs ranging from 100 to 0.625 mg/ml) activities. In vivo antidermatophytic assay demonstrated that clotrimazole cured T. mentagrophytes and M. canis infection on days 21 and 17, respectively, whereas M. communis and C. zeylanicum extracts significantly (p < 0.05) cured T. mentagrophytes and M. canis infection on days 9 and 13 as well as 9, 11, respectively. Phytochemical screening showed the presence of flavonoids, tannins, phenols, and alkaloids in M. communis and alkaloids, flavonoids, and tannins in C. zeylanicum. Findings of the present study also provided the scientific evidence that natural plants could be used in traditional medicine for the prevention and treatment of dermatophytic infections., (© The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Subterranean rodents face unique thermoregulatory challenges. Evaporative water loss (EWL) is a crucial mechanism for maintaining heat balance in endotherms subjected to heat stress but also leads to potential dehydration. EWL depends on gradients of temperature and humidity between the surface of the individual and the surrounding environment. Underground burrows generally provide a stable water vapor saturated atmosphere which may impede evaporative heat loss (EHL). This will mainly occur when ambient temperature exceeds the upper limit of individual's thermoneutral zone, or when body temperature rises as result of digging activities. Here we evaluate the effect of ambient temperature on EWL and energy metabolism in the subterranean rodent Ctenomys talarum (tuco-tucos), which inhabits sealed burrows, but makes an extensive use of the aboveground environment. We observed that EWL is increased when ambient temperature rises above thermoneutrality; below this point, evaporation remains stable. Though EWL contributes to total heat loss by increasing ∼1.3 times at 35°C, dry thermal conductance is raised four times. In tuco-tucos' burrows both non-evaporative and, to some extent, evaporative and behavioral mechanisms are essential for body temperature regulation, preventing overheating at high ambient temperatures in a water vapor-saturated atmosphere., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Hemolytic activity was recently reported for several pathogenic fungal species, such as Aspergillus, Candida, Trichophyton, Penicillium and Fusarium. Based on a number of mechanistic and characterization studies, several fungal hemolysins have been proposed as virulence factors. Hemolysins lyse red blood cells resulting in the release of iron, an important growth factor for microbes especially during infection. The requirement of iron in fungal growth is necessary for metabolic processes and as a catalyst for various biochemical processes. Expression of a hemolytic protein with capabilities to lyse red blood cells has also been suggested to provide a survival strategy for fungi during opportunistic infections. The aims of this study were to investigate the hemolytic activities of dermatophytes species isolated from patients with dermatophytosis. Hair, skin and nail samples of patients were examined with direct microscopy using potassium hydroxide and cultivated on Mycobiotic agar and Sabouraud's dextrose agar. To determine hemolytic activities of dermatophytes species, they were subcultured on Columbia Agar with 5% sheep blood and incubated for 7-14 days at 25°C in aerobic conditions. Media which displayed hemolysis were further incubated for 1-5 days at 37°C to increase hemolytic activity. In this study, 66 dermatophytes strains were isolated from clinical specimens and were identified by six different species: 43 (65.1%) Trichophyton rubrum, 7 (10.7%) Trichophyton mentagrophytes, 5 (7.6%) Microsporum canis, 5 (7.6%) Trichophyton tonsurans, 4 (6.0%) Epidermophyton floccosum and 2 (3.0%) Trichophyton violaceum. Twenty-one T. rubrum strains showed incomplete (alpha) hemolysis and nine T. rubrum strains showed complete (beta) hemolysis, whereas hemolysis was absent in 13 T. rubrum strains. Four T. mentagrophytes strains showed complete hemolysis and three T. tonsurans strains showed incomplete hemolysis. However, M. canis, E. floccosum and T. violaceum species had no hemolytic activity. Hemolytic activity is pronounced in dermatophytes and may play an important role as a virulence factor. Hemolysins produced may play an important role in the balance between the host's cellular immunity and the ability of the fungus to diminish the immune response., (Copyright © 2014 Elsevier Masson SAS. All rights reserved.)

The anamorph of Arthroderma benhamiae is an upcoming zoophilic dermatophyte that only in recent years has gained importance as a cause of tinea in humans. Its identification by conventional methods can cause problems. In this study we have subjected seven genetically confirmed strains of A. benhamiae anamorphs from northern Germany recently identified in our laboratory to a comprehensive assessment. Their macroscopic and microscopic morphology was checked on various agars and enzyme release stimulated by substrates with keratin, hair perforation and other physiological characteristics were tested. All strains were related to the previously described yellow phenotype of the A. benhamiae anamorph and showed a high resemblance among themselves. Coherent features were their uniform thallus morphology on Sabouraud glucose agar with yellow pigmentation, the formation of circuit-like hyphal structures and hyphal connections that had not been described previously, a lack of conidia, thiamine dependence, the spectrum of released enzymes and a good growth on human stratum corneum. With exception of the latter two these criteria are suggested for the identification of this anamorph phenotype that should be evaluated by future observations. Different phenotypes of the A. benhamiae anamorph may prevail in other geographic regions., (© 2014 Blackwell Verlag GmbH.)

Dermatophytes are fungi responsible for a disease known as dermatophytosis. Biofilms are sessile microbial communities surrounded by extracellular polymeric substances (EPS) with increased resistance to antimicrobial agents and host defenses. This paper describes, for the first time, the characteristics of Trichophyton rubrum and T. mentagrophytes biofilms. Biofilm formation was analyzed by light microscopy, scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) as well as by staining with crystal violet and safranin. Metabolic activity was determined using the XTT reduction assay. Both species were able to form mature biofilms in 72 h. T. rubrum biofilm produced more biomass and EPS and was denser than T. mentagrophytes biofilm. The SEM results demonstrated a coordinated network of hyphae in all directions, embedded within EPS in some areas. Research and characterization of biofilms formed by dermatophytes may contribute to the search of new drugs for the treatment of these mycoses and might inform future revisions with respect to the dose and duration of treatment of currently available antifungals.

The most prevalent skin infections are mainly caused by species of dermatophytes of the genera Trichophyton, Microsporum, and Epidermophyton that infect keratinized tissues and stratum corneum of skin and hair. Besides proteases with putative role of kinases and other enzymes, immune modulators are abundantly secreted during infection as well. The molecular mechanism used by the dermatophytes to infect and counteract the host immune response is not well understood. The defense against infections basically depends on the host's immune responses to metabolites of the fungi, virulence of the infecting strain or species and anatomical site of the infection. The two aspects of the immune system, the immediate hypersensitivity and delayed-type hypersensitivity against dermatophytes may be crucial to the progression and severity of skin infection. Management of the infection through species identification and molecular diagnostic techniques as well as use of novel targeted drugs in addition to conventional anti-fungal compounds is of great importance in dealing with disease onsets and outbreaks. Here we reviewed the fungal skin infections elucidating their biologic and immunologic characteristics. Reaction to fungal invasion by the infected epithelial tissue on the host side is also discussed. Moreover, determinants of protective immunity and treatment options are focused that could confer long-lasting resistance to infection.

Hydroxytyrosol (HT) is a potent antioxidant found in olive oil and leaves. Using several in vitro approaches, we tested antifungal activity of HT. HT showed broad spectrum of antifungal activity against medically important yeasts and dermatophyte strains with MIC values ranging between 97.6 µgml⁻¹ and 6.25 mgml⁻¹. The antimicrobial activity of HT was also tested using the time-kill methodology. Below the MIC value, HT showed potent damage of cell wall of Candida albicans ATCC 10231 using fluorescent dye-exclusion method. At the subinhibitory concentration, HT also influenced dimorphic transition of Candida indicating that HT is inhibitor of germ-tube formation as one of the most important virulence factor of C. albicans. Furthermore, HT showed disturbances in cell surface hydrophobicity (CSH) of C. albicans. The in vitro results indicate that HT caused a significant cell wall damage and changes in CSH as well as inhibition of germ-tube formation as virulence factor of C. albicans. The study indicates that HT has a considerable in vitro antifungal activity against medically important yeasts.

Systematic studies about pet guinea pigs with dermatophytosis are rare. The aim of this study was to evaluate clinical signs, therapy and zoonotic risk of pet guinea pigs with dermatophytosis. Questionnaires from both owners (n = 74) of pet guinea pigs with dermatophytosis and their veterinarians (n = 101) were analysed regarding clinical signs, therapy and data pertinent to zoonotic potential. Trichophyton (T.) mentagrophytes was found in 97% of cases. In the weeks preceding the onset of the clinical signs, a new guinea pig joined the household in 43% of cases. One third of the affected guinea pigs had lived in the household for less than 3 months. Predominant clinical signs were alopecia (83%), scaling (73%) and crusting (70%). The most commonly affected body site was the head (75%). In approximately one quarter of the cases humans showed clinical signs of dermatophytosis, in half the households, only children were affected. Skin lesions were seen most often on the face, the neck and the arms. Pet guinea pigs carrying dermatophytes must be considered a serious zoonotic risk for their owners, especially for children. A major risk factor for dermatophytosis seems to be a recent acquisition of a new guinea pig., (© 2012 Blackwell Verlag GmbH.)

Dermatophytes are keratinophilic fungi that can be pathogenic for humans and animals by infecting the stratum corneum, nails, claws or hair. The first infection step consists of adherence of arthroconidia to the stratum corneum. The mechanisms and the kinetics of adherence have been investigated using different in vitro and ex vivo experimental models, most notably showing the role of a secreted serine protease from Microsporum canis in fungal adherence to feline corneocytes. After germination of the arthroconidia, dermatophytes invade keratinised structures that have to be digested into short peptides and amino acids to be assimilated. Although many proteases, including keratinolytic ones, have been characterised, the understanding of dermatophyte invasion mechanisms remains speculative. To date, research on mechanisms of dermatophyte infection focused mainly on both secreted endoproteases and exoproteases, but their precise role in both fungal adherence and skin invasion should be further explored., (© 2011 Blackwell Verlag GmbH.)

The dermatophytes are a group of closely related fungi which are responsible for the great majority of superficial mycoses in humans and animals. Among various potential virulence factors, their secreted proteolytic activity attracts a lot of attention. Most dermatophyte-secreted proteases which have so far been isolated in vitro are neutral or alkaline enzymes. However, inspection of the recently decoded dermatophyte genomes revealed many other hypothetical secreted proteases, in particular acidic proteases similar to those characterized in Aspergillus spp. The validation of such genome predictions instigated the present study on two dermatophyte species, Microsporum canis and Arthroderma benhamiae. Both fungi were found to grow well in a protein medium at acidic pH, accompanied by extracellular proteolysis. Shotgun MS analysis of secreted protein revealed fundamentally different protease profiles during fungal growth in acidic versus neutral pH conditions. Most notably, novel dermatophyte-secreted proteases were identified at acidic pH such as pepsins, sedolisins and acidic carboxypeptidases. Therefore, our results not only support genome predictions, but demonstrate for the first time the secretion of acidic proteases by dermatophytes. Our findings also suggest the existence of different pathways of protein degradation into amino acids and short peptides in these highly specialized pathogenic fungi., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

This study first report to identify the mating type (-)-specific gene of alpha-box and the mating type (+)-specific gene of the high-mobility-group (HMG) DNA-binding domain in zoophilic dermatophytes of Arthroderma benhamiae in an effort to understand the epidemiological characteristics of Trichophyton mentagrophytes. The sequence of the alpha-box gene (1,387 bp) was found to contain two exons, from 184 to 475 bp and from 525 to 1,387 bp, coding a protein of 384 amino acids, beginning with a putative initiating methionine (ATG). The sequence of the HMG gene (1,910 bp) contained two exons, from 234 to 415 bp and from 479 to 1,457 bp, coding a protein of 386 amino acids, beginning with a putative initiating methionine (ATG). PCR analysis detected the alpha-box gene in A. benhamiae (-) mating type strains but not in (+) mating type strains. On the other hand, the HMG gene was detected in A. benhamiae (+) mating type strains but not in (-) mating type strains. These findings suggest that the HMG and alpha-box genes could be specific to the (+) and (-) mating types, respectively.

Thirty-nine Arthroderma vanbreuseghemii isolates identified by mating behavior were assayed to determine their ribosomal RNA gene internal transcribed spacer (ITS) region genotypes. Twenty-two isolates of mating type (+) and 4 of (-) showed the genotype of Trichophyton mentagrophytes var. interdigitale, 9 isolates of mating type (+) and 4 of (-) showed the genotype of A. vanbreuseghemii. Nine of 14 isolates with granular or asteroid colony morphology showed the genotype of T. mentagrophytes var. interdigitale. There was no relation between ITS genotype and morphology or mating type of the isolates. Isolates of T. mentagrophytes var. interdigitale genotype were shown to include non-sexually degenerate zoophilic strains with granular colony morphology as well as sexually and morphologically degenerate anthropophilic strains and may be just one genotype of a heterogeneous species, A. vanbreuseghemii., (Copyright © 2011 The Japanese Society for Medical Mycology)

Dermatophytes are hyphomycetes that can degrade keratin. This puts them in a position to cause infections of the keratin-containing superficial skin. The resulting clinical picture is called tinea. The pathogenesis and course of tinea is decisively determined by pathogen-related factors and by the defense mechanisms of the host. An infection starts with an adherence of fungal propagules, followed by the formation of hyphae that can spread within the tissue. This process is accompanied by a release of fungal enzymes and other pathogenic factors. Next keratinocytes are activated, the epidermal barrier is destroyed, epidermal proliferation is enhanced and defensins are expressed within the epidermis. In addition, innate and specific immune responses are initiated, involving neutrophilic granulocytes, macrophages, antibodies and T cells. The cellular mechanisms are thought to be crucial for healing. Special conditions apply to nail infections, because within nail plates the fungi are not accessible to effective defense mechanisms, as well as to infections of hair follicles that contain specific concentrations of steroid hormones. Dermatophytes that penetrate into the dermis can cause granulomatous inflammatory reactions and systemic immune reactions are supposed to be a trigger of so-called id reactions., (© The Author • Journal compilation © Blackwell Verlag GmbH, Berlin.)

Cutaneous mycoses are among the most common infections in humans and have become an important public health issue because they cause invasive infections in immunocompromised patients. During the infectious process, dermatophyte-host interactions trigger specific metabolic adaptations that allow the pathogen to adhere to and penetrate the host tissue, scavenge nutrients, and overcome the host defense mechanisms. This metabolic shift and the interplay between metabolism, morphogenesis and stress response are important factors that have been extensively studied in several pathogens. Host cells also respond to the pathogen stimuli by activating intracellular signaling pathways that trigger the immune response against the infectious agent. The comprehension of the molecular aspects of these responses may help to establish new therapeutical strategies. In this review, different aspects of the biology of dermatophytes are addressed, with emphasis on the dermatophyte-host interaction and the mechanisms of antifungal resistance.

Crossing of Trichophyton rubrum with Arthroderma simii yielded many ascomata around the fluffy T. rubrum colonies. One of the 35 supposed ascospores isolated from matured ascoma was shown to be a hybrid of the two species. Hybrids were observed within the genotypes of four different genes, i.e., rRNA, actin, DNA topoisomerase II and cytochrome b. These results strongly suggest that T. rubrum is not an asexual or clonal species.

An active compound from the bulb of Eleutherine americana L. Merr. (Iridaceae) collected from East Kalimantan, Indonesia, was tested for its antidermatophyte and antimelanogenesis activity. Antifungal assay-directed fractionation of the n-hexane-soluble fraction of the methanolic extract of the bulb of E. americana led to the isolation of 1 as an active compound. The compound was identified as the naphthoquinone eleutherin by EI-MS and (1)H-, (13)C-, and two-dimensional NMR analyses. Antidermatophyte assay of 1 at concentrations of 10, 20, 40, 60, and 80 microg/disk and myconazole, a commercial antidermatophyte, at 10 microg/disk displayed 7, 8, 13, 16, 17, and 14 mm of inhibition zone against Trichophyton mentagrophytes, respectively. In a melanin formation inhibition assay, compound 1 displayed potent antimelanogenesis activity at 5 ppm with low toxicity compared with arbutin, a commercial skin-whitening agent. The results showed the high potential of 1, an active compound from E. americana, to be applied as an antidermatophyte and antimelanogenesis agent.

Three genetically hybrid F1 progenies produced between a clinical isolate of Arthroderma simii (KMU4810) and a tester strain of A. vanbreuseghemii (RV27961) were crossed with two tester strains of A. vanbreuseghemii (RV27961 and RV27960) and a tester strain of A. benhamiae (RV30001), respectively. Three crossings yielded hybrid second progenies. Another interspecies crossing between A. simii (KMU4810) and a tester strain of A. benhamiae (RV26680) yielded one hybrid F1 progeny (Asb57). The second crossings of F1 progeny (Asb57) with A. vanbreuseghemii (RV27961) and A. benhamiae (RV30001) yielded many hybrid second progenies. Some hybrid second progenies produced between F1 progeny and A. vanbreuseghemii were confirmed to have genes from three species. The gene exchangeability among three Arthroderma species was shown and the meaning of these events discussed.