Trichophyton Rubrum Research Articles

Recalcitrant dermatophyte infections: identification and risk factors.

Recalcitrant dermatophytosis is an emerging phenomenon that occurs worldwide, and Trichophyton indotineae is currently the prominent cause. Skin specimens from patients with tinea infection were obtained by scrubbing and then sectioned into three fragments. Two fragments were subjected to direct microscopic examination and culture, while the third portion was utilized in the PCR method. Isolates were morphologically identified as Trichophyton mentagrophytes/interdigitale complex (n = 60 [83.33%]), Microsporum canis (n = 8 [11.11%]), Trichophyton rubrum (n = 3 [4.16%]), and Epidermophyton floccosum (n = 1 [1.38%]). Among 60 T. mentagrophytes complex isolates, 53 (88.33%) were classified as T. indotineae and seven as T. interdigitale genotype II. The disease duration was longer in the T. indotineae group (P = 0.035). Both Gradient PCR and skin-sampling methods yield similar results in terms of positive and negative cases (P = 1.0000). The time patients stopped their medication did not impact the positive case numbers (P = 0.803). Gender had no effects on the frequency (P = 0.699). Familial contamination, dermatologic disorder, and other underlying conditions did not differ in the two group infections (P > 0.05). Steroid usage is strongly associated with the emergence of tinea infection (P < 0.04). The duration of antifungal administration had a substantial effect on the emergence of resistant organisms (P = 0.05). Steroid usage, T. indotineae involvement, and prolonged exposure to antifungals were the solid and influential factors in recalcitrant involvement. Regarding quick and suitable diagnosis and treatment, which is essential in preventing recalcitrant cases, we suggest that direct skin sample PCR can meet the demands.

Penetration Profile of Terbinafine Compared to Amorolfine in Mycotic Human Toenails Quantified by Matrix-Assisted Laser Desorption Ionization-Fourier Transform Ion Cyclotron Resonance Imaging.

Amorolfine 5% lacquer is an established topical treatment for fungal infection of the nails. The success of topical therapy for onychomycosis depends on whether the permeated drug concentration in the deep nail bed is retained above the effective antifungal minimum inhibitory concentration (MIC). We compared the penetration profile of amorolfine and a new topical formula of terbinafine in human mycotic toenails using matrix-assisted laser desorption ionization mass spectrometry imaging-Fourier transform ion cyclotron resonance (MALDI-FTICR) imaging. Amorolfine 5% lacquer and terbinafine 7.8% lacquer were applied to mycotic nails (n = 17); nail sections were prepared, and MALDI-FTICR analysis was performed. Based on the MICs of amorolfine and terbinafine needed to kill 90% (MIC90) of Trichophyton rubrum, the fold differences between the MIC90 and the antifungal concentrations in the nails (the multiplicity of the MIC90) were calculated overall and for the keratin-unbound fractions. Both amorolfine and terbinafine penetrated the entire thickness of the nail. The mean concentration across the entire nail section 3h following terbinafine treatment was 1414μg/g of tissue (equivalent to 4.9mM) compared with 780μg/g (2.5mM) following amorolfine treatment (not significantly different; p = 0.878). The median multiplicity of the MIC90 was significantly higher in amorolfine- than terbinafine-treated nails overall (191 vs. 48; p = 0.010) and for the keratin-unbound fractions only (7.4 vs. 0.8; p = 0.002). In this ex vivo study, MALDI-FTICR demonstrated that, although amorolfine 5% and terbinafine 7.8% had similar distribution profiles, both penetrating from the surface to the nail bed, the concentration of amorolfine in the nail was significantly higher than that of terbinafine relative to their respective MIC90 values. Clinical studies are required to determine whether these effects translate to a clinical difference in treatment success.

The StuA Transcription Factor and Alternative Splicing Mechanisms Drive the Levels of MAPK Hog1 Transcripts in the Dermatophyte Trichophyton rubrum.

Trichophyton rubrum is a human fungal pathogen that causes dermatophytosis, an infection that affects keratinized tissues. Integrated molecular signals coordinate mechanisms that control pathogenicity. Transcriptional regulation is a core regulation of relevant fungal processes. Previous RNA sequencing data revealed that the absence of the transcription factor StuA resulted in the differential expression of the MAPK-related high glycerol osmolarity gene (hog1) in T. rubrum. Here we validated the role of StuA in regulating the transcript levels of hog1. We showed through RT-qPCR that transcriptional regulation controls hog1 levels in response to glucose, keratin, and co-culture with human keratinocytes. In addition, we also detected hog1 pre-mRNA transcripts that underwent alternative splicing, presenting intron retention in a StuA-dependent mechanism. Our findings suggest that StuA and alternative splicing simultaneously, but not dependently, coordinate hog1 transcript levels in T. rubrum. As a means of preventing and treating dermatophytosis, our results contribute to the search for new potential drug therapies based on the molecular aspects of signaling pathways in T. rubrum.

Penetration Profiles of Four Topical Antifungals in Mycotic Human Toenails Quantified by Matrix-Assisted Laser Desorption Ionization-Fourier Transform Ion Cyclotron Resonance Imaging.

Onychomycosis is a fungal infection of the nails that can be challenging to treat. Here, matrix-assisted laser desorption ionization-Fourier transform ion cyclotron resonance (MALDI-FTICR) imaging was applied to the quantitative analysis of the penetration profile of the antifungal compound, amorolfine, in human mycotic toenails. The amorolfine profile was compared with those of three other antifungals, ciclopirox, naftifine, and tioconazole. Antifungal compounds (amorolfine 5% lacquer, ciclopirox 8% lacquer, naftifine 1% solution, and tioconazole 28% solution) were applied to mycotic nails (n = 42). Nail sections were prepared, and MALDI-FTICR analysis was performed on the sections at a spatial resolution of 70 μm to compare the distribution profiles. Based on the minimum inhibitory concentrations of the four test compounds needed to kill 90% (MIC90) of the fungal organism, Trichophyton rubrum, the fold differences between the MIC90 and the antifungal concentrations in the nails (termed the multiplicity of the MIC90) were calculated for each. The penetration profiles indicated higher concentrations of amorolfine and ciclopirox in the deeper layers of the nails 3h after treatment, compared with naftifine and tioconazole. The mean concentrations across the entire nail sections at 3h were significantly different among the four antifungals: amorolfine, 2.46mM; ciclopirox, 0.95mM; naftifine, 0.63mM; and tioconazole, 1.36mM (p = 0.016; n = 8 per compound). The median multiplicity of the MIC90 at 3h was 191-fold for amorolfine, tenfold for ciclopirox, 52-fold for naftifine, and 208-fold for tioconazole. In this study, MALDI-FTICR was successfully applied to the quantitative analysis of antifungal distribution in human mycotic nails. The findings suggest that amorolfine penetrates deeper layers of the nail and accumulates at concentrations far exceeding the MIC needed to exert antimycotic activity.

Chemical composition of Artemisia argyi essential oil and its antifungal activity against dermatophytes by inhibiting oxidative phosphorylation and causing oxidative damage

Ethnopharmacological relevanceDermatophytes are notorious pathogens capable of infecting various mammals skin, posing serious threats to human health and overall life quality worldwide. Artemisia argyi has been recorded and applied for over a thousand years to treat skin itching. Although it has the potential to be developed as a plant-based antifungal agent, it’s antifungal activity and action mechanism of active ingredients are still unclear. Aim of the studyThe aim of this study was to investigate the chemical composition, antifungal activity against skin fungi, and potential mechanisms of Artemisia argyi essential oil (AEO). Materials and MethodsThe chemical composition of AEO was analyzed by gas chromatography-mass spectrometry (GC-MS) firstly. Flat growth restraint and double half dilution tests was performed to evaluate AEO antifungal activity against Microsporum gypseum, Trichophyton mentagrophytes, and Trichophyton rubrum. And then, the physiological mechanism of AEO inhibiting dermatophytes was systematically explored through scanning electron microscopy, relative conductivity, membrane leakage, ROS content, and antioxidant enzyme activity. Finally, the main pathways were screened through transcriptome sequencing, while the related genes expression levels and enzyme activity were validated. ResultsMonoterpenes and sesquiterpenoids were the most highly representative class of AEO. AEO had powerful antifungal activity against M. gypseum, T. mentagrophytes, and T. rubrum, with minimum inhibitory concentration (MIC) values of 0.6, 1.2, and 1.2 μL/mL, respectively. Moreover, AEO can also damage the cell membrane integrity of T. mentagrophytes, resulting in cellular extravasation of intracellular substances. Transcriptome analysis revealed that the main target of AEO is to inhibit electron transfer and oxidative phosphorylation during respiration, ultimately leading to obstruction of normal ATP synthesis and energy metabolism in mitochondria. And a large amount of ROS will generate due to the incompletely catalysis of oxygen under mitochondrial complexes. Coupled with the decrease of antioxidant enzyme (SOD, POD) activity, excessive accumulation of ROS will cause serious oxidative damage to cells and eventually exhibiting antifungal activity against dermatophytes. ConclusionsThe present study demonstrated that Artemisia argyi was a valuable source of active compounds with antifungal activity. These findings support AEO as a potential agent to inhibit dermatophytes and prevent related dermatophytoses.

PH- and Matrix Metalloproteinase-Responsive Multifunctional Bilayer Microneedles Platform for Treatment of Tinea Pedis.

Persistent foot odor and itchiness are common symptoms of tinea pedis, significantly disrupting the daily life of those affected. The cuticular barrier at the site of the tinea pedis is thickened, which impedes the effective penetration of antifungal agents. Additionally, fungi can migrate from the skin surface to deeper tissues, posing challenges in the current clinical treatment for tinea pedis. To effectively treat tinea pedis, we developed a platform of bilayer gelatin methacrylate (GelMA) microneedles (MNs) loaded with salicylic acid (SA) and FK13-a1 (SA/FK13-a1@GelMA MNs). SA/FK13-a1@GelMA MNs exhibit pH- and matrix metalloproteinase (MMP)-responsive properties for efficient drug delivery. The MNs are designed to deliver salicylic acid (SA) deep into the stratum corneum, softening the cuticle and creating microchannels. This process enables the antibacterial peptide FK13-a1 to penetrate through the stratum corneum barrier, facilitating intradermal diffusion and exerting antifungal and anti-inflammatory effects. In severe cases of tinea pedis, heightened local pH levels and MMP activity further accelerate drug release. Our research demonstrates that SA/FK13-a1@GelMA MNs are highly effective against Trichophyton mentagrophytes, Trichophyton rubrum, and Candida albicans. They also reduced stratum corneum thickness, fungal burden, and inflammation in a guinea pig model of tinea pedis induced by T. mentagrophytes. Furthermore, it was discovered that SA/FK13-a1@GelMA MNs exhibit excellent biocompatibility. These findings suggest that SA/FK13-a1@GelMA MNs have significant potential for the clinical treatment of tinea pedis as well as other fungal skin disorders.

In vitro Growth of Trichophyton Rubrum in the Presence of 80% Ethanolic Extract of Grapefruit Pepins as Antifungal Agent

Introduction: Many modern antifungal drugs have limits in terms of germ resistance, and to remedy this, plant compounds are being explored to find new more effective principles. Aim: With the aim of complementing the efforts of modern medicine against viral, bacterial, parasitic and fungal diseases, the plant extract of 80% hydroalcoholic grapefruit seed extract of Citrus paradisi was tested on the in vitro growth of Trichophyton rubrum. Methods: Antifungal tests were carried out on Sabouraud medium, to which the plant extracts were incorporated using the double dilution method in inclined tubes. A 10 µL volume of the Trichophyton rubrum suspension was inoculated on the culture medium contained in the test tubes. Results: Results showed that Trichophyton rubrum was sensitive to the 80% hydroethanolic extract of Citrus paradisi in a dose-dependent manner. Conclusion: The hydroethanolic extract may be a source for the development of Traditional Improved Medicines (TIM) against skin mycosis.

Clinical Characteristics and Pathogen Spectrum of Male Genital Fungal Infections in Nanchang Area, South China.

The cutaneous fungal infections in male genitalia are relatively rare, and often present with various atypical clinical symptoms. It was mainly reported in a small number of case reports, while data with large number of patients were rarely reported. In this study, we reported 79 male patients with cutaneous fungal infections on scrotum or penis. The fungal infections were confirmed by microscopic examination directly and fungus culture. Clinical characteristics and predisposing factors were also collected. Of these 79 patients, 72 has lesions on scrotum, 5 on penis and 2 on both scrotum and penis. Trichophyton (T.) rubrum is the most common pathogen, found in 50 (67.6%) patients, which presented diverse clinical manifestation such as majorly erythematous, dry diffused scaly lesions without a clear border, slightly powdery and scutular scalings. Candida (C.) albicans is the secondly common pathogen, found in 21 (28.4%) patients, which also presented diverse lesions such as erythematous with dry whitish scaly lesions and erythematous erosion. The predisposing factors mainly included concomitant fungal infections on sites other than genitalia, especially inguinal region (tinea cruris), application of corticosteroid and high moisture. In conclusion, cutaneous fungal infections in male genitalia could be caused by different fungi, showed atypical or mild clinical appearances in most cases and might be a fungus reservoir, emphasizing the necessity to timely perform the fungi examinations and corresponding therapy.

Phytochemical Composition, Antioxidant and Antifungal Activity of Essential Oils and Crude Extracts of &lt;em&gt;Dittrichia viscosa maritime&lt;/em&gt; (L.), an Aromatic and Medicinal Plant from Northern Morocco

Dittrichia viscosa, a perennial herb belonging to the Asteraceae family, was collected in a coastal site located in northern Morocco. This study focused on exploring the phytochemical composition and biological activities of essential oils (EOs) and solvents (methanol and acetone) extract of leaves, stems, and flowers. Total phenolic (TPC) and total flavonoid (TFC) contents were determined, and the result showed a high level of TPC and TFC in leaves with the values of 127.61 mg GAEs/g and 117.82 mg CEs/g, respectively. HPLC-MS analysis highlighted 18 phenolic acids and flavonoids, with notable quantities of apigenin 7-glucoside (2233 μg/g) and chlorogenic acid (1429 μg/g). GC‒MS analysis of EOs allowed the identification of 27 biologically active compounds, dominated by decanoic acid (20.98%) and toluene (19.66%). Regarding the biological activities, antioxidant activity was evaluated through the DPPH test, while antifungal activity was tested using the microdilution method. The most potent antioxidant effect was observed in the methanol extract of leaves, with a value of 0.18 mg/mL, while the weakest effect was noted in the acetone extract of flowers, registering a value of 1.63 mg/mL. Similarly, the antioxidant activity of methanolic extracts surpassed that of acetonic extracts in each part of D. viscosa. Concerning antifungal activity, the findings reveal that all extracts demonstrated important capacity against the tested pathogens. The methanolic extract displayed high efficacy compared to the acetonic one, with MFC values ranging between 1.94-3.38 mg/mL and 2.88-4.75 for Candida albicans, Aspergillus niger and Trichophyton rubrum, respectively. However, for Fusarium oxysporum f. sp. albedinis, the MFCs were higher (7.5-8.5 mg/mL). Regarding EOs, the MFCs were more favorable for the three human pathogenic fungi, ranging between 1.88 and 3.35 µL/mL, compared to F. oxysporum, which has an MFCs of 3.5 µL/mL. In summary, the findings demonstrated that D. viscosa could be considered a useful alternative and reliable source of bioactive compounds for the pharmaceutical and agricultural industries.

The Ptk2-Pma1 pathway enhances tolerance to terbinafine in Trichophyton rubrum.

The increasing prevalence of dermatophyte resistance to terbinafine, a key drug in the treatment of dermatophytosis, represents a significant obstacle to treatment. Trichophyton rubrum is the most commonly isolated fungus in dermatophytosis. In T. rubrum, we identified TERG_07844, a gene encoding a previously uncharacterized putative protein kinase, as an ortholog of budding yeast Saccharomyces cerevisiae polyamine transport kinase 2 (Ptk2), and found that T. rubrum Ptk2 (TrPtk2) is involved in terbinafine tolerance. In both T. rubrum and S. cerevisiae, Ptk2 knockout strains were more sensitive to terbinafine compared with the wild types, suggesting that promotion of terbinafine tolerance is a conserved function of fungal Ptk2. Pma1 is activated through phosphorylation by Ptk2 in S. cerevisiae. Overexpression of T. rubrum Pma1 (TrPma1) in T. rubrum Ptk2 knockout strain (ΔTrPtk2) suppressed terbinafine sensitivity, suggesting that the induction of terbinafine tolerance by TrPtk2 is mediated by TrPma1. Furthermore, omeprazole, an inhibitor of plasma membrane proton pump Pma1, increased the terbinafine sensitivity of clinically isolated terbinafine-resistant strains. These findings suggest that, in dermatophytes, the TrPtk2-TrPma1 pathway plays a key role in promoting intrinsic terbinafine tolerance and may serve as a potential target for combinational antifungal therapy against terbinafine-resistant dermatophytes.

Rosemary and neem: an insight into their combined anti-dandruff and anti-hair loss efficacy

Dandruff, a common scalp disorder characterized by flaking dead skin, is often treated with conventional topical products. However, limitations exist due to potential side effects and high costs. Therefore, searching for natural, cost-effective solutions for dandruff and hair loss is crucial. Rosemary herb and neem tree, both cultivated in Egypt, possess well-documented anti-inflammatory properties derived from their rich phenolic phytoconstituents. This study formulated a standardized combined extract of rosemary and neem (RN-E 2:1) into hair gel and leave-in tonic formats. This extract demonstrated superior efficacy against Malassezia furfur (a causative agent of dandruff) and Trichophyton rubrum (associated with scalp disorders) compared to the conventional antifungal agent, ketoconazole. The combined extract (RN-E 2:1) also exhibited potent anti-inflammatory activity. Additionally, the suppression of iNOS expression is considered concentration-dependent. Quality control verified formulation stability, and ex-vivo studies confirmed effective ingredient penetration into the epidermis, the primary site of fungal presence. Remarkably, both formulations outperformed the standard treatment, minoxidil in hair growth trials. These findings highlight the potential of natural extracts for scalp and hair health.

Prevalence and risk factors for onychomycosis in acute care dermatology wards.

We conducted a cross-sectional study on the clinical and mycological features of onychomycosis in patients in the dermatology ward of Iwate Medical University Hospital, an acute care hospital. Of the 226 hospitalized patients, 73 (32.3%) had onychomycosis and 61 (26.9%) were diagnosed after admission. The toenail was the most common site of onychomycosis (94.5%), while toenail plus fingernail and fingernail only sites were 4.1% and 1.4%, respectively. The most common clinical form of onychomycosis was distal and lateral subungual onychomycosis (79%) with Trichophyton rubrum (66.7%) and T. interdigitale (27.8%) as the main causative species. Patients who were older, or had neurological diseases, or needed stretcher transfer had onychomycosis significantly more frequently than those who were obese, had diabetes, cancer, needed an escort for moving, or could move independently. Our study suggests that there is likely to be a significant number of untreated and undiagnosed patients with onychomycosis in acute care hospitals. Therefore, it is necessary to increase awareness of onychomycosis in hospitals.

Isolation, prevalence of bacteria and fungi from carcasses and different locations within Yola abattoir and antimicrobial resistance profile of Escherichia coli O157:H7

The absence of modern slaughtering facilities and an unhygienic environment in Yola abattoir may result in an outbreak of antibiotic-resistant pathogenic microorganisms, leading to economic loss and human death. Hence, this study was carried out to isolate and assess the prevalence of bacteria and fungi within the abattoir environment as well as to determine the antimicrobial resistance profile of E. coli O157:H7 from different locations and carcasses. A cross-sectional study was conducted where a simple random sampling method was used for sampling. A total of 156 samples were collected. Isolation and identification of bacteria and fungi were done using standard methods. Pseudomonas was the most dominant bacterial species with a total of 151 isolates, followed by E. coli 143 and Klebsiella with 123 isolates. The highest prevalence in fungi was recorded in Aspergillus flavus at 17.6 %, followed by Aspergillus niger at 15.1 % and Trichophyton rubrum and Penicillium that had 14.5 % respectively. The highest mean air-borne coliform count was in the Lairage, it ranged between 1.1 x 106 CFU/m3 and 1.7 x 106 CFU/m3. A hundred and forty-three E. coli were isolated out of which six (4.2 %) were characterized to be E. coli O157:H7 producing characteristic colorless colonies on cefixime tellurite sorbitol MacConkey (CT-SMAC) agar. All the six E. coli O157:H7 were resistant to at least two antibiotics, ampicillin, and amoxicillin-clavulanic acid while ciprofloxacin, trimethoprim-sulfamethoxazole, and tetracycline were effective against the six E. coli O157:H7. The MAR ranged between 0.2 and 0.7. The results obtained in this study showed that there is a need for modern facilities in Yola abattoir, training of workers on hygiene, and a hygienic environment to avoid microbial contamination and spread of antimicrobial-resistant pathogenic strains like E. coli O157:H7 and mycotoxin producer Aspergillus flavus.

Clinicomycological Study of Dermatophyte Infections in Patients Attending Dermatology Outpatient Department in a Tertiary Care Hospital

Context: Dermatophytosis is a fungal disease characterized by the infection of skin, hair, and nails. There is an upsurge of complicated superficial dermatophytosis and an increase in the incidence of chronic, relapsing, recurrent cases in India that are also often unresponsive to conventional doses of drugs. Aims: The aim of this study was to study the prevalence of dermatophyte species in the particular study area and it is susceptibility to itraconazole, terbinafine, and fluconazole. Settings and Design: All dermatophytes isolates obtained were subjected to 10% KOH mount, fungal culture, and antifungal sensitivity testing for fluconazole, itraconazole, and terbinafine. Materials and Methods: Newly diagnosed cases of dermatophytosis without treatment for the past 3 months were included in the study. KOH preparation was observed and subjected to fungal culture. Then, the susceptibility of the dermatophytes to oral antifungals (itraconazole, terbinafine, and fluconazole) was done. Results: Out of 28 patients with tinea, 14 males and 12 females were tested KOH positive. Out of them, 23 patients showed positive for dermatophyte growth. In them, 15 isolates were Trichophyton mentagrophytes and 8 isolates were Trichophyton rubrum. Antifungal drug susceptibility testing data indicated that, for the two species, MIC of itraconazole ranged between 0.03 and 16 μg/ml, terbinafine between 0.03 and 16 μg/ml, and fluconazole between 0.25 and 64 μg/ml. Conclusions: The study provided information regarding the prevalent species of dermatophyte in our study area and it is high sensitivity to itraconazole compared to terbinafine and the lowest for fluconazole. This gives us guidance as to which antifungal to use empirically in our clinical practice to treat tinea infection.

Extraction of Synephrine from Waste Peels of Citrus sinensis and Green Synthesis of Gold Nanoparticles from it against Dermatophytes

The main object of the current work was to determine the antifungal efficiency of secondary metabolites product called synephrine that extracted from Citrus sinesis peels and the ability of synephrine to biosynthesis gold nanoparticles from HAucl4 which consider environmentally favourable method, then determine their activity against pathogenic human dermatophyte. The identification of synephrine done by Thin layer chromatography (TLC), High Performance Liquid Chromatography (HPLC) and The Fourier Transform Infrared (FTIR). The characterization of gold nanoparticles by using Ultra Violet-Visible Spectroscopy (UV-Vis), Field – Emission Scanning Electron Microscopy (FESEM) and Fourier Transform Infrared (FTIR), confirmed the biosynthesis of gold nanoparticles in diameter and morphology for AuNps biosynthesis by C. sinensis was 9.7-31 (nm) rounded to oval shape. The synephrine and AuNps that formed use it against some dermatophytes Trichophyton mentographytes, Trichophyton rubrum and Microsporum canis, the activity of synephrine against T. mentographytes at (10, 15 and 20 mg/mL) give less inhibition effect as compare with antifungal effect, while M. canis in 15 mg/mL show best effect than antifungal and for gold nanoparticles most concentration effective was (20 mg/mL).

Antifungal Activity of Trichoderma orientale FMR 12486 Crude Extract against Some Human Pathogenic Fungi

     This work aimed to investigate the prevalence of pathogenic fungi and evaluate the antifungal activity of Trichoderma orientale FMR12486 crude extract against pathogenic fungi isolated from patients attending the National Center for Thoracic and Respiratory Diseases (having a history of tuberculosis) and consultant of Dermatology of Baghdad hospital, Iraq. A total of 80 clinical specimens were collected: 20 skin scrapings specimens and 60 sputum specimens. The results of direct examination by KOH 10% and culture showed that 11 (55%) cases from 20 skin specimens were positive for fungal infections, while in the sputum specimens, 28 (47%) cases from 60 were positive. Candida albicans represented the most common fungal infection isolated from sputum specimens which represented 18 cases at a rate of 64.2%, followed by Candida tropicalis 5 (17.9%), Aspergillus fumigates 2 (7.1%), Aspergillus flavus 1 (3.6%), Candida glabrata 1 (3.6%) and Candida krusei 1 (3.6%). While in the skin scrapes, Candida parapsilosis was the common which represented 3 cases at a rate of 27.3%, followed by Trichophyton mentagrophyte 2 (18.2%), Microsporium canis 2 (18.2%), C. albicans 2 (18.2%), Trichophyton rubrum 1 (9.1%), C. tropicalis 1 (9.1%). T. orientale isolates were collected from the gardens soil of the University of Baghdad and were then identified depending on the morphological feature of the colony and microscopic characteristics. To confirm identification, PCR technique was used in which DNA of T. orientale was extracted at a concentration of 54 - 294 μg/2 g wet mycelium with a purity of 1.6 - 1.8. DNA samples were amplified with primers ITS-1and ITS-4, And the results confirmed that the isolate was T. orientale strain FMR12486. This isolate was processed with ethyl acetate using a standard extraction method and the final amount of crude extract being 6 grams. Different concentrations of crude extract 0.5, 1, 2 and 4 mg/ml were prepared and used against fungal pathogens isolated using the agar-well diffusion method. The results showed high antifungal activity against pathogenic isolates, significantly recording maximum inhibition zones of 20 and 22 mm against C. parapsilosis at concentrations of 0.5 and 1 mg/ml, respectively. It was followed by T. mentagrophyte and C. glabrata. 16, 20, and 16, 20 mm that were recorded at concentrations of 2 and 4mg/ml, a significant maximum inhibition zone was recorded 26 mm against C. glabrata, followed by C. parapsilosis, M. canis and T. mentagrophyte (24, 26; 16, 24 and 20, 24mm respectively). Just Trichophyton rubrum fungus in this study showed resistance to all concentrations of the crude extract where the inhibition zone was zero. The results of the current study showed that T. orientale strain FMR12486 crude extract can be used to make an effective drug to treat human fungal infections.

Dermatophytes and skin dermatophytoses in Southeast Asia-First epidemiological survey from Cambodia.

Dermatomycoses count to the most frequent dermatoses in Cambodia. The aim of this survey was to investigate the occurrence of dermatophytes in this Southeast Asian country. From June 2017 to July 2018, skin scrapings were taken from 67 patients with superficial dermatophytosis for mycological diagnostics. Identification of dermatophytes was confirmed by sequencing of the 'internal transcribed spacer'-(ITS) region of the rDNA, and the gene of the Translation Elongation Factor (TEF)-1α. Patients were suffering from tinea corporis and tinea inguinalis/cruris 42/67 (63%), tinea capitis/faciei 14/67 (21%), tinea corporis/capitis/faciei 6/67 (9%), tinea manuum/pedis 2/67 (3%), tinea pedis 2/67 (3%) and tinea manuum 1/67 (1%). Both, by culture and/or PCR, a dermatophyte was detected in 52 (78%) out of 67 samples. Culture positive were 42 (81%) of 52, PCR positive were 50 (96%). The following dermatophytes were found: Trichophyton (T.) rubrum, 36/52 strains (69%, 29 by culture), T. mentagrophytes/T. interdigitale (TM/TI) 9/52 (17%, six by culture) and Microsporum (M.) canis 5/52 strains (10%, by culture). One strain of Nannizzia (N.) incurvata 1/52 (2%) and N. nana 1/52 (2%) was isolated. Based on sequencing, we demonstrated that two T. mentagrophytes strains out of the nine TM/TI represented the new ITS genotype XXV Cambodia. We found one T. mentagrophytes strain genotype VIII (now, reclassified as T. indotineae). This isolate was terbinafine resistant, and it exhibited the amino acid substitution Phe397Leu in the squalene epoxidase. Three strains of T. interdigitale genotype II* were isolated. This is the first survey on epidemiology of dermatophytes in Cambodia. Currently, T. rubrum represents the most frequent species in Cambodia. One Indian strain genotype VIII T. mentagrophytes was found. A highlight was the first description of the new T. mentagrophytes genotype XXV Cambodia.

Ozone Therapy on Fungi that Cause Onychomycosis

ABSTRACT Onychomycosis is a common fungal infection of the nails, predominantly caused by dermatophytes and is a notoriously difficult condition to treat. Thus, the objective of the present study was to evaluate and compare, in vitro, the efficacy of different applied ozone dosages against the main fungi that cause onychomycosis. Dermatophyte fungi Trichophyton rubrum (ATCC 28,188) and Microsporum gypseum (ATCC 24,102) were incubated at 28 °C for 14 days. Treatments were divided into nine experimental groups according to applied ozone dosages: Control Group 0 mg/m2: fungi were cultured but received no treatment; cultured fungi that received ozone therapy for 157, 314, 471, 628, 785 943, 1100 and 1257 mg/m2. The main results indicate that topical ozone therapy effectively reduced the germination percentage of T. rubrum and M. gypseum in all experimental periods, achieving a complete eradication with a treatment time of 628 and 1257 mg/m2, respectively. In conclusion, ozone therapy used topically with an applied ozone dosage of 628 mg/m2 for T. rubrum and 1257 mg/m2 for M. gypseum promoted antifungal action on the main dermatophytes responsible for critical complications of onychomycosis. Therefore, ozone therapy can be proposed as an adjuvant for dermatological treatments

Evaluation of the antifungal properties of nanoliposomes containing rhinacanthin-C isolated from the leaves of Rhinacanthus nasutus.

Fungal infections represent a challenging threat tothe human health. Microsporum gypseum and Trichophyton rubrum are pathogenic fungi causing various topical mycoses in humans. The globally emerging issue of resistance to fungi demands the development of novel therapeutic strategies. In this context, the application of nanoliposomes as vehicles for carrying active therapeutic agents can be a suitable alternative. In this study, rhinacanthin-C was isolated from Rhinacanthus naustus and encapsulated in nano-liposomal formulations, which were prepared by the modified ethanol injection method. The two best formulations composed of soybean phosphatidylcholine (SPC), cholesterol (CHL), and tween 80 (T80) in a molar ratio of 1:1:0 (F1) and 1:1:0.5 (F2) were proceeded for experimentation. The physical characteristics and antifungal activities were performed and compared with solutions of rhinacanthin-C. The rhinacanthin-C encapsulating efficiencies in F1 and F2 were 94.69 ± 1.20% and 84.94 ± 1.32%, respectively. The particle sizes were found to be about 221.4 ± 13.76 nm (F1) and 115.8 ± 23.33 nm (F2), and zeta potential values of -38.16 mV (F1) and -40.98 mV (F2). Similarly, the stability studies of rhinacanthin-C in liposomes demonstrated that rhinacanthin-C in both formulations was more stable in mediums with pH of 4.0 and 6.6 than pure rhinacanthin-C when stored at the same conditions. Rhinacanthin-C in F1 was slightly more stable than F2 when stored in mediums with a pH of 10.0 after three months of storage. However, rhinacanthin-C in both formulations was less stable than pure rhinacanthin-C in a basic medium of pH 10.0. The antifungal potential was evaluated against M. gypsum and T. rubrum. The findings revealed a comparatively higher zone of inhibition for F1. In the MIC study, SPC: CHL: T80 showed higher inhibition against M. gypseum and a slightly higher inhibition against T. rubrum compared to free rhinacanthin-C solution. Moreover, rhinacanthin-C showed significant interaction against 14α-demethylase in in silico study. Overall, this study demonstrates that nanoliposomes containing rhinacanthin-C can improve the stability and antifungal potential of rhinacanthin-C with sustained and prolonged duration of action and could be a promising vehicle for delivery of active ingredients for targeting various fungal infections.

New thymol-derived triazole exhibits promising activity against Trichophyton rubrum.

Fungal infections have emerged worldwide, and azole antifungals are widely used to control these infections. However, the emergence of antifungal resistance has been compromising the effectiveness of these drugs. Therefore, the objective of this study was to evaluate the antifungal and cytotoxic activities of the nine new 1,2,3 triazole compounds derived from thymol that were synthesized through Click chemistry. The binding mode prediction was carried out by docking studies using the crystallographic structure of Lanosterol 14α-demethylase G73E mutant from Saccharomyces cerevisiae. The new compounds showed potent antifungal activity against Trichophyton rubrum but did not show relevant action against Aspergillus fumigatus and Candida albicans. For T. rubrum, molecules nº 5 and 8 showed promising results, emphasizing nº 8, whose fungicidal and fungistatic effects were similar to fluconazole. In addition, molecule nº 8 showed low toxicity for keratinocytes and fibroblasts, concluding that this compound demonstrates promising characteristics for developing a new drug for dermatophytosis caused by T. rubrum, or serves as a structural basis for further research.