Common Dermatophyte Species Research Articles

Enhanced treatment in cutaneous dermatophytosis management by Zataria multiflora-loaded nanostructured lipid carrier topical gel: A randomized double-blind placebo-controlled clinical trial

The global rising incidence of skin mycosis especially dermatophytes currently affects more than 20–25% of the world population. It is believed that nanotechnology can aid in the efficient treatment of this skin disease. The present study, therefore, aimed to employ nanostructured lipid carriers (NLCs) in a gel formulation for a more effective delivery of Zataria multiflora (ZM) essential oils through the skin to manage mild to moderate cutaneous dermatophytosis. Zataria multiflora-loaded nanostructure lipid carriers (ZM-NLCs) were prepared and optimized by utilizing an ultrasonic probe approach and formulated the ZM-NLCs as a 1% w/w carbopol gel after confirming the NLCs-related characteristics. In vitro antifungal susceptibility testing was performed on 20 common dermatophyte species based on the CLSI M38-A3 guidelines. A total of 80 volunteers (40 volunteers from each gender, equally divided into two groups: ZM-NLCs gel and placebo receivers) participated in this clinical randomized, double-blind placebo-controlled study. Clinical manifestations (itching, scaling, size of lesion and inflammation), as well as mycological findings such as KOH-direct examination test and culture, were evaluated after 2 and 4 weeks of topical application. Molecular identification of the strains isolated from the patient's lesions was also performed to the species level using a PCR-RFLP method. A mono-dispersed suspension of spherical nanoparticles with zeta potential, Z-average and PDI index of −26.6 ± 7.7 mV, 273.5 ± 4 nm and 0.33 ± 0.03, respectively was achieved with no cytotoxicity. Apart from the significant inhibitory effect of ZM-NLCs on fungal growth, the application of the ZM-NLCs gel resulted in an effective diminution in inflammation (87.5%), itching (90%) and scaling (95%) among the volunteers after 4 weeks. The aforementioned signs/symptoms were significant when compared to the patients receiving a placebo (p < 0.005). A reduction/disappearance of the lesion was also significantly reported in the ZM-NLCs gel receivers. Compared with placebo receivers, meaningful negative results for both direct examination and culture were observed (17.5% vs 70% for KOH-direct examination and 12.5% vs 57.5% for culture) for patients who received ZM-NLCs gel after 2 weeks of prescription. PCR-RFLP outputs revealed T. mentagrophytes/interdigitale complex as the predominant isolated species from cutaneous lesions. In conclusion, Zataria multiflora-NLCs gel application displayed a more rapid cure time with no adverse side effects compared to placebo demonstrating the efficacious nature of NLCs.

Factors influencing epidemiology of dermatomycoses at Gdansk, Poland and Grodno, Belarus.

IntroductionAn increase in numbers of cutaneous fungal infections is being observed around the world. Dermatomycoses occur in every age group and can significantly decrease quality of life. Examining the prevalence and epidemiological trends of cutaneous fungal infections is necessary for developing new effective preventive, diagnostic and therapeutic methods. Many factors can influence the growth and patterns of global spread of the different species of fungi.AimTo investigate the differences between the prevalence of dermatophytes and cutaneous fungal infections observed at two mycological laboratories and their plausible causes.Material and methodsMycological examination was performed in 7324 patients in Gdansk, Poland and in 4729 patients in Grodno, Belarus who exhibited clinical signs and symptoms of cutaneous fungal infections. Direct preparations was made in 20% KOH with 40% DMSO. Additionally cultures were prepared on modified Sabouraud dextrose agar.ResultsDermatophytes were cultured in 642 (53.58%) cases in Gdansk. The most common species of dermatophytes were Trichophyton rubrum (306 cases), Trichophyton mentagrophytes var. granulosum (193 cases) and Microsporum canis (127 cases). Yeasts were isolated in 531 (44.40%) cases out of which Candida albicans was the most frequently identified in 323 cases. Similarly, dermatophytes were the most common in 1158 (68.72%) cases in Grodno. Candida were isolated in 527 (31.28%) cases. Analysing the localization of dermatomycoses onychomycosis was observed in majority of the infected patients in Gdansk. In Grodno the most common location of superficial cutaneous infection was tinea corporis In the Gdansk region dermatomycoses were observed in 688 female patients and in 508 male patients. In Grodno the prevalence is inverted with the majority of cases observed in 921 males and 764 females.ConclusionsThe prevalence of superficial fungal infections is higher in Grodno, Belarus when compared to Gdansk, Poland. Similarly, dermatophytes were most commonly observed in both Laboratories.

Tinea Capitis by Microsporum canis in an Elderly Female with Extensive Dermatophyte Infection

Tinea capitis is a type of dermatophyte infection primarily affecting children. We report a case of an elderly woman with well-controlled diabetes mellitus presenting with a six-month history of erythema with yellow crusts on her scalp and extensive erythematous patches with scales on the body skin. She adopted a stray cat before the disease onset. Dermoscopic findings and manifestation under the Wood’s lamp favoured the diagnosis of tinea capitis. Further microscopic examinations of her scalp, including direct KOH and fluorescence stain examination, fungal culture and polymerase chain reaction sequencing identification confirmed the diagnosis of tinea capitis caused by Microsporum canis. Treatment with oral terbinafine was effective. Adult tinea capitis is often misdiagnosed due to its rarity and atypical presentation. However, in some regions, the incidence of tinea capitis in immunocompetent adults is rising which requires the awareness of clinicians. A thorough history (including the animal contacting history), physical examination and further mycological examinations are required for diagnosis. Trichophyton violaceum is the most common dermatophyte species in most regions while adult tinea capitis caused by Microsporum canis is less common. Terbinafine, griseofulvin and itroconazole have been reported to be effective drugs for the treatment of tinea capitis, and terbinafine can be considered as systemic treatment in elderly patients with comorbidities to reduce the drug–drug interaction.

In Vitro Antifungal Activity of Green Synthesized Silver Nanoparticles in Comparison to Conventional Antifungal Drugs Against Trichophyton Interdigitale, Trichophyton Rubrum and Epidermophyton Floccosum.

Dermatophytosis is a globally distributed fungal infection. Treatment failure and relapse is common in this disease. Silver nanoparticles are known for their promising antimicrobial activity. The aim of this study was to determine the antifungal activity of these nanoparticles against common dermatophyte species. A set of 30 molecularly identified dermatophytes including Trichophyton interdigitale (n=10), Trichophyton rubrum (n=10), and Epidermophyton floccosum (n=10) were used in this study. Green synthesized silver nanoparticles using chicory (Cichorium intybus) were tested for their antifungal activity in comparison to fluconazole, itraconazole and terbinafine. Interspecies differences in minimum inhibitory concentrations of antifungal drugs and silver nanoparticles were tested using Kruskal-Wallis test in SPSS software version 21. The highest minimum inhibitory concentrations (MICs) among antifungal drugs were observed for fluconazole [range: 4-64 μg/mL, geometric mean (GM) =17.959 μg/mL], followed by itraconazole (range: 0.008-0.5, GM= 0.066) and terbinafine (range: 0.004-0.25 μg/mL, GM=0.027 μg/mL). Silver nanoparticles showed potent antifungal activity against all dermatophyte isolates with MICs (range: 0.25-32 μg/mL, GM=4.812 μg/mL) higher than those of itraconazole and terbinafine, but lower than fluconazole. MIC values of silver nanoparticles demonstrated significant differences between species (P=0.044), with E. floccosum having the highest MICs (GM=9.849 μg/mL) compared to T. interdigitale (GM=3.732 μg/mL) and T. rubrum (GM=3.031 μg/mL). Silver nanoparticles demonstrated promising anti-dermatophyte activity against the studied dermatophytes. Due to their wide-spectrum activity against other fungal and bacterial pathogens, they could be a potential choice, at least in the case of cutaneous and superficial infections.

Clinico-mycological Evaluation of Dermatophytes in a Tertiary Care Hospital of Saurashtra, Gujarat, India

Introduction: Dermatophytes and opportunistic fungi are common skin infection mainly affecting superficial layers of integument with rare systemic involvement. The higher prevalence in India is supposedly due to the hot and humid climatic condition, occupation and low socio-economic status. Recent upsurge in fungal infections are due to increased incidence of immunosuppressive state. Aim: To study the clinical manifestation and mycological evaluation of dermatophytosis. Materials and Methods: The present cross-sectional observational study was conducted during December 2007 to October 2009 in MP Shah Medical College and GG Hospital, Jamnagar, Gujarat, India. Samples were taken from 200 patients with clinically diagnosed dermatophytosis. Depending on the site of lesion, samples from skin, hair or nail were taken. These samples were than examined directly under microscope and cultured on Sabouraud’s Dextrose Agar (SDA) and Cornmeal Agar (CMA). Descriptive statistics like mean and percentages were used to infer results. Results: Out of 200 patients, 177 (88.5%) samples were positive by Potassium Hydroxide (KOH) mount while 127 (63.5%) samples were positive by culture. Most common species of dermatophytes identified was Trichophyton rubrum followed by Trichophyton mentagrophytes. Agricultural workers were found to be more prone to dermatophytic infection and males (61%) were more commonly affected than females (39%). Conclusion: Dermatophytosis is a common superficial mycotic infection in Saurashtra region where hot and humid climate, poor socio-economic condition and compromised personal hygiene play an important role in fungal growth. In the present study, tinea corporis was found to be the most common clinical type with T. rubrum being the commonest isolated species.

The first succinylome profile of Trichophyton rubrum reveals lysine succinylation on proteins involved in various key cellular processes

BackgroundDermatophytes, the most common cause of fungal infections, affect millions of individuals worldwide. They pose a major threat to public health because of the severity and longevity of infections caused by dermatophytes and their refractivity to therapy. Trichophyton rubrum (T. rubrum), the most common dermatophyte species, is a promising model organism for dermatophyte research. Post-translational modifications (PTMs) have been shown to be essential for many biological processes, particularly in the regulation of key cellular processes that contribute to pathogenicity. Although PTMs have important roles, little is known about their roles in T. rubrum and other dermatophytes. Succinylation is a new PTM that has recently been identified. In this study, we assessed the proteome-wide succinylation profile of T. rubrum. This study sought to systematically identify the succinylated sites and proteins in T. rubrum and to reveal the roles of succinylated proteins in various cellular processes as well as the differences in the succinylation profiles in different growth stages of the T. rubrum life cycle.ResultsA total of 569 succinylated lysine sites were identified in 284 proteins. These succinylated proteins are involved in various cellular processes, such as metabolism, translation and epigenetic regulation. Additionally, 24 proteins related to pathogenicity were found to be succinylated. Comparison of the succinylome at the conidia and mycelia stages revealed that most of the succinylated proteins and sites were growth-stage specific. In addition, the succinylation modifications on histone and ribosomal proteins were significantly different between these two growth stages. Moreover, the sequence features surrounding the succinylated sites were different in the two stages, thus indicating the specific recognition of succinyltransferases in each growth phase.ConclusionsIn this study, we explored the first T. rubrum succinylome, which is also the first PTM analysis of dermatophytes reported to date. These results revealed the major roles of the succinylated proteins involved in T. rubrum and the differences in the succinylomes between the two major growth stages. These findings should improve understanding of the physiological and pathogenic properties of dermatophytes and facilitate future development of novel drugs and therapeutics for treating superficial fungal infections.

Analysis of zoonotic dermatophytoses in 64 family-based groups

Objective To investigate the distribution and epidemiology of fungal pathogens in zoonotic dermatophytoses. Methods Seventy-four patients with dermatophytoses and 72 pets from 64 families, who were all culture positive for dermatophytes, were included in this study and classified into 64 family-based groups. Fungal culture and direct microscopic examination were carried out for species identification of fungal isolates, internal transcribed spacer（ITS）sequence analysis and random amplified polymorphic DNA（RAPD）analysis were performed for molecular identification and homology analysis. Results Dermatophyte species were consistent among the patients and pets from the same families for all the 64 family-based groups. A total of 146 fungal strains were isolated, including 93 Microsporum canis（M. canis）strains and 53 Trichophyton interdigitale（T. interdigitale）strains. M. canis was isolated from 42（65.7%）family-based groups including 34 groups keeping cats and 8 groups keeping dogs, while T. interdigitale from 22（34.3%）groups, including 14 groups keeping rabbits, 6 groups keeping cats and 2 groups keeping dogs. There were 54（75.0%）pets with obvious clinical symptoms（erythema, desquamation, depilation, etc）, and 18（25.0%）asymptomatic pets which were all cats. Among the 18 asymptomatic cats, 14 carried M. canis, and 4 T. interdigitale. ITS sequencing and RAPD analysis revealed a high homology between the fungal pathogens in the same family-based groups. Conclusions M. canis and T. interdigitale are common species of dermatophytes in zoonotic dermatophytoses, and both of them have host specificity. Zoonotic dermatophytes can be transmitted between human and domestic animals, and attention should be paid to asymptomatic animals（carriers）. Key words: Tinea; Microsporum; Trichophyton; Pets; Random amplified polymorphic DNA technique; Zoonoses

Laser irradiation on growth of trichophyton rubrum: an in vitro study.

Trichophyton rubrum is one of the most common species of dermatophytes which affects superficial keratinous tissue. It is not especially virulent but it can be responsible for considerable morbidity. Although there are different therapeutic modalities to treat fungal infections, clinicians are searching for alternative treatment because of the various side effects of the present therapeutic methods. As a new procedure, Laser therapy has brought on many advantages in clinical management of dermatophytes. Possible inhibitory potential of laser irradiation on fungal colonies was investigated invitro in this study. A total of 240 fungal plates of standard size of trichophyton rubrum colonies that had been cultured from the lesions of different patients at the mycology laboratory, were selected. Each fungal plate was assigned as control or experimental group. Experimental plates were irradiated by a laser system (low power laser or different wavelength of high power laser). The effects of different laser wavelengths and energies on isolated colonies were assessed. After laser irradiation, final size of colonies was measured on the first, the 7th and the 14th day after laser irradiation. Although low power laser irradiation did not have any inhibitory effect on fungal growth, the Q-Switched Neodymium-Doped Yttrium Aluminium Garnet (Nd:YAG) laser 532nm at 8j/cm(2), Q-Switched Nd:YAG laser 1064nm at 4j/cm(2) to 8j/cm(2) and Pulsed dye laser 595nm at 8j/cm(2) to 14j/cm(2) significantly inhibited growth of trichophyton rubrum in vitro. Q-Switched Nd:YAG 532nm at 8j/cm(2), Q-Switched Nd:YAG laser 1064nm at 4j/cm(2) to 8j/cm(2) and pulsed dye laser (PDL) 595nm at 8j/cm(2) to 14j/cm(2) can be effective to suppress trichophyton rubrum growth.

Tinea capitis in Botswana

BackgroundTinea capitis (TC) is a common dermatophyte infection of the scalp that can also involve the eyebrows and eyelashes.AimThis study aimed to find the causative fungus responsible for TC in Botswana and determine its association with the clinical types of TC.MethodsSamples for potassium hydroxide 10% mounts and fungal cultures were collected in a microbiology laboratory at the National Health Laboratory, Gaborone, Botswana. Dermasel agar and Sabouraud dextrose agar were inoculated with the samples. Lactophenol cotton blue mounts were prepared from the culture-positive samples to study the morphological characteristics.ResultsTrichophyton violaceum was found to be the predominant causative organism of TC. Trichophyton tonsurans was isolated from one patient. Both are anthropophilic species.ConclusionTC was found to be most common in those aged 1–15 years (81%). Of 17 patients in this age group, 16 were younger than 10 years old and one was 14 years old. T. violaceum was the most common dermatophyte species isolated.

Species analysis of fungi isolated in a laboratory from patients with mycoses during 1960-2006

Objective To analyze fungal isolates from patients with superficial fungal infections during 1960-2006.Methods Fungal strains isolated from patients with superficial (mucocutaneous and cutaneous)fungal infections and identified in the Medical Mycology Clinical Laboratory,Department of Dermatology and Venereology,Union Hospital,from 1960 to 2006 (data from September 1991 to July 1992 were unavailable),were subjected to a classification and statistical analysis.Clinical samples for mycological examination were taken from outpatients or inpatients of different departments in hospitals of Hubei province and surrounding areas.Morphological,physiological and biochemical methods were applied for species identification.Results A total of 11 989 Candida strains were isolated,which belonged to 23 species and 16 genera.They fell into 3 groups,i.e.,dermatophytes,Candida and yeasts (including Malassezia),and non-dermatophyte moulds.Since 287 strains of moulds were suspected to be contaminating fungi,11 702 residual isolates were analyzed.Of the analyzed isolates,Candida species (5642/11 702,48.2％ )and dermatophytes (5279/11 702,45.1％ )predominated,followed by yeasts (449/11 702,3.8％) and Malassezia species (332/11 702,2.8％).The most frequently isolated species was Trichophyton rubrum (3865/11 702,33.0％),Candida albicans (3110/11 702,26.6％ ) and non-albicans Candida species (2532/11 702,21.6％ ).Dermatophyte strains were mostly isolated from lesions of smooth skin with an exception of palmoplantar and interdigit regions (1787/5279,37.7％).The most common dermatophyte species was Trichophyton rubrum,followed by Trichophyton violanceum.Candida was mainly isolated from mucous membrane lesions (4099/5642,72.7％),with Candida albicans being the predominant species.Conclusions Candida species and dermatophytes predominate in patients with superficial fungal infections during 1960-2006,with Trichophyton rubrum being the most common species. Key words: Dermatomycoses; Epidermaphyton; Candida

Naftifine demonstrates potent fungicidal activity against the most common dermatophyte species with no evidence of resistance

Naftifine demonstrates potent fungicidal activity against the most common dermatophyte species with no evidence of resistance

The Sensitivity of Dermatophytes to the Antimicrobial Activity of Manuka Honey and Other Honey

Honey has been reported to have antifungal activity and so was tested against clinical isolates of the common dermatophyte species which cause tineas in man. A honey with an average level of hydrogen peroxide, and a manuka (Leptospermum scoparium J. R. and G. Forst, family Myrtaceae) honey with an average level of non-peroxide antibacterial activity were used. An agar well diffusion assay was used, the contents of the wells being replaced with freshly prepared honey solutions at 24-h intervals over the 3–4 days of incubation. The lowest concentrations (% v/v, in steps of 5%) of manuka honey with catalase added to remove hydrogen peroxide, and of the other honey (without catalase) showed that inhibition of growth around the wells were, respectively, Epidermophyton floccosum 25%, 10%; Microsporum canis 25%, 15%; Microsporum gypseum 55%, 20%; Trichophyton mentagrophytes var. interdigitale 45%, 15%; Trichophyton mentagrophytes var. mentagrophytes 25%, 15%; Trichophyton rubrum 20%, 5% and Trichophyton tonsurans 25%, 20%. No inhibitory activity was detected with the other honey at 50% (v/v) with catalase added. The results of this investigation show that the common dermatophytes are sensitive to the antimicrobial activity of honey, indicating that clinical evaluation of honey in the treatment of tineas is warranted. This would determine whether the hydrogen peroxide or the non-peroxide antifungal agent diffuses better into the skin.

Direct detection of five common dermatophyte species in clinical samples using a rapid and sensitive 24-h PCR-ELISA technique open to protocol transfer

Identification of dermatophytes is usually based on morphological characteristics determined by time-consuming microscopic and cultural examinations. An effective PCR-ELISA method has been developed for rapid detection of dermatophyte species directly from clinical specimens within 24 h. Isolated genomic DNA of skin scrapings and nail samples from patients with suspected dermatophyte infections is amplified with species-specific digoxigenin-labelled primers targeting the topoisomerase II gene. The subsequent ELISA procedure with biotin-labelled probes allows a sensitive and specific identification of the five common dermatophytes -Trichophyton rubrum, T. interdigitale, T. violaceum, Microsporum canis and Epidermophyton floccosum. PCR-ELISA, based on the new polyphasic species concept, was assessed using 204 microscopy-positive samples in two university mycological laboratories in Munich and Tübingen, and 316 consecutive specimens - regardless of mycological findings - in a dermatological practice laboratory in Neu-Ulm. One of the five dermatophytes was confirmed by PCR-ELISA in 163 of 204 (79.9%) of the clinical samples from the university hospitals found positive using microscopy. Culture was positive for dermatophytes in 59.8% of the same cases. A significant difference between these two methods could be demonstrated using the McNemar test (P < 0.005). Analysis of specimens from Neu-Ulm confirmed the results in a dermatological practice laboratory as 25.0% of the specimens had positive PCR results, whereas only 7.3% were positive according to culture. Direct DNA isolation from clinical specimens and the PCR-ELISA method employed in this study provide a rapid, reproducible and sensitive tool for detection and discrimination of five major dermatophytes at species level, independent of morphological and biochemical characteristics.

Molecular typing ofTrichophyton mentagrophytesvar.interdigitaleisolated in a university hospital in Japan based on the non‐transcribed spacer region of the ribosomal RNA gene

Detecting intraspecies polymorphisms in fungi causing dermatophytoses is important in elucidating routes of infection and determining whether Tinea recurrence is caused by exacerbation or re-infection. In fungi, the non-transcribed spacer region (NTS) of the ribosomal RNA gene shows the greatest accumulation of base sequence mutations. We therefore assessed NTS sequences in 64 clinical isolates of Trichophyton mentagrophytes var. interdigitale, the second most common species of dermatophytes in Japan. These isolates were among the clinical isolates of dermatophytes in the Department of Dermatology, Kanazawa Medical University Hospital in 2006 and were obtained by morphological and molecular biological identification methods. DNA was extracted from each isolate, as well as from one isolate maintained in our department, to detect length polymorphisms at each of three variable loci, TmiS0, TmiS1 and TmiS2, of the NTS for subtyping. We observed seven patterns for TmiS0, six patterns for TmiS1 and three patterns for TmiS2. The combinations of these patterns enabled us to classify the 65 isolates into 15 types. The most prevalent, constituted 46% (30/65) of all isolates. Eleven types were new combinations, whereas the other four were previously described. These results suggest that this method may be used to determine the molecular epidemiology of T. mentagrophytes var. interdigitale in Japan, because it generated results rapidly and in a sensitive manner.

Molecular identification of dermatophytes isolated in Sfax-Tunisia

Recently, molecular techniques have been developed for rapid and accurate alternative identification of dermatophyte. The objective was to investigate PCR sequencing for identification of common dermatophyte species. Twenty-four samples were taken from patients with dermatophytosis. Mycological identification revealed: Trichophyton rubrum (13), Trichophyton mentagrophytes (seven), Microsporum sp. (two) and negative culture (two). PCR sequencing of chitin synthase1 gene from culture was applied for 17 strains and directly from samples for seven cases. PCR amplification of CHS1 gene gave a 450 bp fragment. Sequencing showed 97 to 99% identity. Identification of our strains by conventional methods was confirmed by PCR sequencing in 83.3% of cases. Two isolates of Microsporum sp. were reclassified as M. ferrugineum. In the two cases with negative culture, direct PCR sequencing has detected T. mentagrophytes. PCR sequencing provided excellent tool for identifying strains that do not present typical morphological characteristics. It was also able to give correct identification of the first isolates of M. ferrugineum in Tunisia. Récemment, les techniques moléculaires ont été développées comme des alternatives rapides et précises d’identification des dermatophytes. Notre objectif a été d’appliquer le séquençage de produits PCR pour l’identification des espèces de dermatophytes les plus fréquentes. Vingt-quatre échantillons ont été prélevés chez des patients atteints de dermatomycoses. L’identification mycologique a révélé : Trichophyton rubrum (13), Trichophyton mentagrophytes (sept), Microsporum sp. (deux) et culture négative (deux). Le séquençage d’amplicons PCR du gène de la « chitine synthase 1 » à partir de la culture a été appliqué pour 17 souches et directement à partir du prélèvement pour sept cas. L’amplification par PCR du gène CHS1 a permis d’obtenir un fragment de 450 pb. Le séquençage a montré 97 à 99 % d’identité. L’identification de nos souches par les méthodes conventionnelles a été confirmée par PCR séquençage dans 83,3 % des cas. Deux isolats de Microsporum sp. ont été reclassés comme M. ferrugineum. Dans les deux cas de culture négative, le séquençage de produits PCR appliqué directement à partir des squames a détecté T. mentagrophytes. Le séquençage de produits PCR a fourni un excellent outil pour l’identification des souches qui ne présentent pas des caractéristiques morphologiques typiques. Il a été aussi capable de donner l’identification correcte des premiers isolats de M. ferrugineum en Tunisie.

TrED: the Trichophyton rubrum Expression Database

BackgroundTrichophyton rubrum is the most common dermatophyte species and the most frequent cause of fungal skin infections in humans worldwide. It's a major concern because feet and nail infections caused by this organism is extremely difficult to cure. A large set of expression data including expressed sequence tags (ESTs) and transcriptional profiles of this important fungal pathogen are now available. Careful analysis of these data can give valuable information about potential virulence factors, antigens and novel metabolic pathways. We intend to create an integrated database TrED to facilitate the study of dermatophytes, and enhance the development of effective diagnostic and treatment strategies.DescriptionAll publicly available ESTs and expression profiles of T. rubrum during conidial germination in time-course experiments and challenged with antifungal agents are deposited in the database. In addition, comparative genomics hybridization results of 22 dermatophytic fungi strains from three genera, Trichophyton, Microsporum and Epidermophyton, are also included. ESTs are clustered and assembled to elongate the sequence length and abate redundancy. TrED provides functional analysis based on GenBank, Pfam, and KOG databases, along with KEGG pathway and GO vocabulary. It is integrated with a suite of custom web-based tools that facilitate querying and retrieving various EST properties, visualization and comparison of transcriptional profiles, and sequence-similarity searching by BLAST.ConclusionTrED is built upon a relational database, with a web interface offering analytic functions, to provide integrated access to various expression data of T. rubrum and comparative results of dermatophytes. It is devoted to be a comprehensive resource and platform to assist functional genomic studies in dermatophytes. TrED is available from URL: .

In vitro activity of Citrus bergamia (bergamot) oil against clinical isolates of dermatophytes

Recently, bergamot oil was shown to be a potent antifungal agent in vitro against clinically important Candida species. In this study, the activities of bergamot natural essence and its furocoumarin-free and distilled extracts on dermatophytes such as Trichophyton, Microsporum and Epidermophyton species were investigated. In vitro susceptibility testing assays on 92 clinical isolates of dermatophytes (Trichophyton mentagrophytes n = 20, Trichophyton rubrum n = 18, Trichophyton interdigitale n = 15, Trichophyton tonsurans n = 2, Microsporum canis n = 24, Microsporum gypseum n = 1 and Epidermophyton floccosum n = 12) were performed using the CLSI M38-A broth microdilution method, except for employing an inoculum of 1-3 x 10(3) cfu/mL. MICs were determined at a visual endpoint reading of 80% inhibition compared with the growth control. MICs (v/v) of all fungi ranged from 0.156% to 2.5% for the natural essence, from 0.02% to 2.5% for the distilled extract, and from 0.08% to 1.25% for the furocoumarin-free extract. The three isolates of T. tonsurans and M. gypseum exhibited the highest MIC values. Data from this study indicate that bergamot oil is active in vitro against several common species of dermatophytes, suggesting its potential use for topical treatment of dermatophytoses.

Specific Primers for Rapid Detection ofMicrosporum audouiniiby PCR in Clinical Samples

This report describes application of PCR fingerprinting to identify common species of dermatophytes using the microsatellite primers M13, (GACA)4, and (GTG)5. The initial PCR analysis rendered a specific DNA fragment for Microsporum audouinii, which was cloned and sequenced. Based on the sequencing data of this fragment, forward (MA_1F) and reverse (MA_1R) primers were designed and verified by PCR to establish their reliability in the diagnosis of M. audouinii. These primers produced a singular PCR band of 431 bp specific only to strains and isolates of M. audouinii, based on a global test of 182 strains/isolates belonging to 11 species of dermatophytes. These findings indicate these primers are reliable for diagnostic purposes, and we recommend their use in laboratory analysis.

Identification of common species of dermatophytes by PCR-RFLP

To establish a simple, sensitive and effective technique for the identification of six common dermatophytes, polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) targeting Topoisomerase II gene were used. The DNA of 6 common dermatophytes was amplified by primer dPsD1 and then primers dPsD2. The products generated by dPsD2 were digested with restriction enzyme Hinc II and Hinf I separately. A DNA fragment of about 3390 bp was amplified by using primer dPsD1 from the genomic DNA of each dermatophyte species. The product of dPsD2 was 2380 bp and the restriction profiles of Hinc II and Hinf I were between 58-1670 bp. By using PCR-RFLP, all of the 6 dermatophytoses were diagnosed to species level and no obvious difference identification between Hinc II and Hinf I. It is concluded that the PCR-RFLP identification of dermatophytes by Hinc II or Hinf I is efficient and rapid in clinical practice.

Clinical identification of common species of dermatophytes by PCR and PCR-RFLP

To find a fast and efficient way of identifying seven common dermatophytes in clinical practice, we used the techniques of polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) targeting Topoisomerase II gene. The DNA of 7 dermatophytes, along with Candida albicans, Aspergillus terreus and Aspergillus flavus were amplified by consensus primer dPsD1. They were then subjected to a second PCR with primers dPsD2 and species-specific primers PsT and PsME separately. 6 of the products generated by dPsD2 were digested with restriction enzyme Hinc II. DNA fragments of 3390 bp and 2380 bp was amplified by using consensus primer dPsD1 and dPsD2 from the genomic DNA of each dermatophyte species separately. By combining the results of the two species-specific primer sets (PsT and PsME), all species of dermatophyte yielded unique sizes-set of PCR products expect for T. mentagrophytes and T. tonsurans. From the restriction profiles of Hinc II, 6 of the 7 dermatophytoses were diagnosed to species level including T. mentagrophytes and T. tonsurans. By combining the results of the PCR and PCR-RFLP, the 7 common dermatophytes can be identified to species level. It is conclude that the multiplex PCR and PCR-RFLP identification targeting the DNA topoisomerase II gene is rapid and efficient.