Evaluation of the Multiplex Real-Time PCR DermaGenius® Assay for the Detection of Dermatophytes in Hair Samples from Senegal

For the successful treatment of dermatophytoses, especially tinea capitis, there is a need for accurate and rapid diagnostic methods. A lot of recent literature has focused on the detection of dermatophytes directly on sample material such as nails, hair and skin scrapings. Molecular tools offer the ability to rapidly diagnose dermatophytosis within 48 h. This study aimed to compare the results of a commercial real-time PCR (real-time PCR) assay DermaGenius®(DG) 2.0 complete multiplex kit with those of conventional diagnostic methods (direct microscopy and culture). A total of 129 hair samples were collected in Dakar (Senegal) from patients suspected of dermatophytosis. DG was applied for the molecular detection of Candida albicans, Trichophyton rubrum/soudanense, T. interdigitale, T. tonsurans, T. mentagrophytes, T. violaceum, Microsporum canis, M. audouinii, Epidermophyton floccosum, T. benhamiae and T. verrucosum. Dermatophytes species and C. albicans were differentiated by melting curve analysis. The sensitivity and specificity of the PCR assay were 89.3% and 75.3%, respectively. DG PCR was significantly more sensitive than culture (p < 0.001). DG PCR is fast and robust to contamination. In this paper, the main questions discussed were the replacement of culture by a broad-spectrum fungal real-time PCR and the implementation of DG PCR into a routine laboratory in Senegal.

Absence of Reported cases of Microsporum ferrugineum, Trichophyton violaceum, and Trichophyton schoenleinii in South Korea

Dermatophytosis is a skin disorder caused by dermatophytes. Dermatophytes isolated in South Korea include Trichophyton (T.) rubrum, T. mentagrophytes, T. verrucosum, T. tonsurans, T. violaceum, T. schoenleinii, Microsporum (M.) canis, M. ferrugineum, M. gypseum, and Epidermophyton floccosum. T. tonsurans was first found in South Korea in 1992. In contrast, there have been no recent reported cases of T. violaceum, T. schoenleinii, and M. ferrugineum in South Korea. Population mobility, changes in human lifestyles, development of the healthcare system, and the introduction of antifungals have brought about dermatophyte evolution in the skin microenvironment. We have reviewed the cases of dermatophytosis caused by M. ferrugineum, T. violaceum, and T. schoenleinii reported both in South Korea and globally.

A simple multiplex polymerase chain reaction assay for rapid identification of the common pathogenic dermatophytes:Trichophyton interdigitale, Trichophyton rubrum, and Epidermophyton floccosum

Background and Purpose: The most common etiological agents of human dermatophytosis in various parts of the world are Trichophyton rubrum, Trichophyton interdigitale, and Epidermophyton floccosum. The main aim of this study was to design and evaluate a simple and straightforward multiplex polymerase chain reaction (PCR)assay for reliable identification/differentiation of these species in clinical isolates. Materials and Methods: The reliable sequences of several molecular targets of dermatophytes species were used to design a multiplex PCR for the identification of common pathogenic dermatophytes. The isolates and clinical specimens examined in this study included seven standard strains of dermatophytes, 101 isolates of dermatophytes and non-dermatophyte molds/yeasts which had already been identified by sequencing or PCR-restriction fragment length polymorphism (RFLP), and 155 clinical samples from patients suspected of cutaneous mycoses. Results: Species-specific primer pairs for T. rubrum and T. interdigitale/T. mentagrophytes were designed based on the sequence data of the translation elongation factor 1-alpha gene, and the primers for E. floccosum targeted the specific sequence of the internal transcribed spacer region (ITS). The multiplex PCR successfully detected T.rubrum, T. interdigitale/T. mentagrophytes, and E. floccosum strains that were identified by sequencing or PCR-RFLP. However, the primer pairs selected for T. interdigitale/T. mentagrophytes cross-reacted with Trichophyton tonsurans. In testing the PCR system directly for clinical samples, the proportion of positive multiplex PCR was higher than positive culture (68.1% vs. 55.4%, respectively). Conclusion: The multiplex assay could detect three common agents out of several causal agents of dermatophytosis, namely T. rubrum, T. interdigitale, and E. floccosum.Therefore, by adding pan-dermatophyte primers it can be used as a comprehensive detection/identification test.

Antioxidant, antiradical power of Syzygium aromaticum essential oil, and its antidermatophytic activity against Epidermophyton floccosum and Trichophyton soudanense

In order to provide an effective alternative for efficient management of dermatophytosis, we evaluate in vitro the antioxidant and antiradical potentials of the essential oil of the dry flower buds of Syzygium aromaticum and its antidermatophytic activity against Epidermophyton floccosum and Trichophyton soudanense, For this purpose, the essential oil was obtained by hydrodistillation through a Clevenger apparatus and the antioxidant and antiradical potentials were evaluated by the ferric reducing antioxidant power (FRAP) and the trapping of the ABTS•+ radical methods, respectively. The evaluation of the antidermatophytic activity was made by the agar incorporation method. The results showed that the essential oil reduced ferric iron (Fe3+) to ferrous iron (Fe2+) in a more significant manner than BHT (reference antioxidant). Moreover, the antiradical power of the essential oil was twice greater than that of BHT. Furthermore, the essential oil inhibited the mycelial growth of the two dermatophytes, with 500 ppm and 1000 ppm of minimal inhibitory concentrations against Epidermophyton floccosum and Trichophyton soudanense, respectively. This activity was greater than that of griseofulvin whose minimal inhibitory concentration was greater than 4000 ppm against both studied germs. These findings show that, withon the framework of safeguarding human lives and protecting the environment, the essential oil from the dry flower buds of Syzygium aromaticum appeared as reliable alternative for the treatment of dermatophytosis caused by Epidermophyton floccosum and Trichophyton soudanense. Our results lay scientific foundation toward the promotion and development of Cameroonian biodiversity in treatments of dermatophytosis.

Comparative Efficacy of Antidermatophyte Creams Formulated from Ethanol Extracts of Cassia Occidentalis and Cassia Alata Leaves

Incidence of drug resistance by dermatophytes is a worldwide concern. Researchers search for natural and better drugs to arrest the ugly situation. This study aims to formulate antidermatophyte Creams with ethanol extracts of the leaves of cassia alata and cassia occidentalis against four selected dermatophytes namely Trichophyton mentagrophtes, Microsporum audounii, Epidermophyton floccosum and Malassezia furfur. Wistar rats were used in determining the efficacy of the formulated herbal creams by examining the biopsies of the skin of the infected rats for presence of fungal hyphae, tissue destruction, Inflammation of the skin, absence of sebaceous gland and also absence of hair follicles. Dekeratinization of the skin layer was also measured. From the result obtained, it was observed that the efficacy of the creams is concentration dependent. Cassia alata cream formulation was particularly effective against three of the dermatophytes namelyTrichophyton mentagrophtes (30.71± 0.51 µm) Microsporum audounii (36.17± 0.59 µm), and Epidermophyton floccosum (37.49± 0.18 µm) while Cassia occidentalis cream was effective against Malassezia furfur (33.22± 0.16 µm). The results showed that the plants ethanol extracts can be utilized in the management of dermatophytosis when formulated as a cream for topical use.

Synthesis of (E)-1-(2,4-Bis((1-phenyl-1H-1,2,3-triazol-4-yl)methoxy)phenyl)- 3-phenylprop-2-en-1-one Derivatives and their Antimicrobial Activity

A new series of chalcone based bis 1,4-disubstituted 1,2,3-triazole heterocyclic molecules (6a-j) were synthesized via the reaction of 1-(2,4-bis((1-phenyl-1H-1,2,3-triazol-4-yl)methoxy)phenyl)ethanone (6a-e) with substituted benzaldehyde. All the molecules were characterized by IR, 1H and 13C NMR and MS spectra spectral data. Compounds 6a-j were evaluated for their antimicrobial effects using a broth disc diffusion method. The compounds were evaluated for their in vitro antimicrobial potential against Gram-positive (Micrococcus luteus, Methicillin-resistant Staphylococcus aureus, Bacillus subtilis and Bacillus cereus), Gram-negative (Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli and Proteus vulgaris) bacterial and fungal (Microsporum canis, Microsporum gypseum and Epidermophyton floccosum) strains by comparison of the reference drugs, zentamycin sulphate (antibacterial) and nystatin (antifungal). The screened compounds 6f, 6e, 6c and 6i were found to be the most active against all the tested bacterial and fungal strains.

Antidermatophyte Activity of Ethanol Extract and Stability of Creams Formulated from Cassia alata Linn

There are scientific reports of medicinal plants having curative properties. These properties may be due to the presence of various phytochemicals in the plants. Cassia alata is a specie in the Fabaceae family. The purpose of this study is to determine the antidermatophyte activity of the extract, formulate a cream with the extract and determine the stability of the cream by measuring the pH, Free-thaw test, Centrifugation and sensitivity to light. We also subjected the formulated cream to temperature variation test at -10, 4, 30, 37 and 45o C. The percentage yield was 8.5%. The extract contain alkaloid (4.24±0.24%), saponin(1.35±0.39%), tannin (0.45±0.29%) and flavonoid (2.42±0.32%) respectively. The antifungal test results showed the activity against the dermatophytes to be in the increasing order M.furfur (12± 0.2 mm) < Microsporum audounii (12±0.4 mm) < Trichophyton mentagrophtes (14±0.2mm) < Epidermophyton floccosum (16±0.2 mm).Temperature stability and Centrifuge testing indicated that the formulations were stable. Light testing indicated no change in the colour of the cream. Our study showed that the plant has high potential as an anti-dermatophyte when formulated as a cream for topical use. The stability of the cream formulations indicated that it can be used for the management of dermatophytosis.

Antidermatophytic activity of some newly synthesized arylhydrazonothiazoles conjugated with monoclonal antibody

A new series of 5-arylhydrazonothiazole derivatives 5a–d has been synthesized, elucidated, and evaluated for their antidermatophytic activity. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the newly synthesized products were investigated against 18 dermatophyte fungal isolates related to Epidermophyton floccosum, Microsporum canis, and Trichophyton rubrum. The morphological alterations induced by the synthesized derivatives singly or conjugated with the monoclonal antibody were examined on spores of T. rubrum using a scanning electron microscope. The efficacy of synthesized derivative 5a applied at its respective MFC alone or conjugated with anti-dermatophyte monoclonal antibody 0014 in skin infection treatment of guinea pigs due to inoculation with one of the examined dermatophytes, in comparison with fluconazole as standard reference drug was evaluated. In an in vivo experiment, the efficiency of 5a derivative conjugated with the antibody induced 100% healing after 45 days in the case of T. rubrum and M. canis-infected guinea pigs.