Dermatophyte Species Research Articles

Molecular Methods For Diagnostics Of Dermatomycoses – Review Of Available Techniques And Evaluation Of Their Advantages And Disadvantages In Implementation For In Routine Use

Abstract Fungal infections of the skin, hairs, and nails undeniably dominate among all types of fungal infections. The etiological factors of the majority of superficial fungal infections are dermatophytes which, although they are the oldest microorganisms considered as pathogens, have long been unstable in the taxonomic position. From a diagnostic point of view, the species identification of dermatophytes is still a serious problem, often generating therapeutic errors. An increasing number of infections, including zoonoses, lack of taxonomic stability and ambiguous clinical picture of all cases of dermatomycosis induce to search for new, fast, repeatable and at the same time cheap methods of species identification of these fungi. In the last decade, revolutionary progress has been observed in the development of molecular methods for the diagnosis of fungal infections and the reliable identification of species of etiological factors that cause these dermatomycoses. The results of many studies indicate that the direct identification of fungi from dermatological samples based on molecular methods is much more reliable and much faster compared to that carried out by conventional methods. Often, the etiological factor of the observed changes was also identified, while the result of cultivation was negative. Particular molecular methods used in the species identification of fungi directly from the clinical material differ in the procedures of genomic DNA extraction, PCR techniques used, the molecular marker used and the results interpretation system. This paper reviews literature regarding different methods of diagnosing of superficial fungal infections based on molecular biology techniques, their advantages and limitations, as well as critical factors for their implementation for routine use. The position of microbiologists in this matter seems to be a foregone conclusion, the time when molecular diagnostics will replace the conventional techniques, based on the cultivation of dermatophytes and assessing their morphology, inexorably coming. Molecular methods of identifying aetiological factors of dermatomycoses directly from dermatological samples are much more attractive and have many advantages. 1. Introduction. 2. Importance of identification of dermatophyte species in dermatological samples. 3. Molecular species identification in pure dermatophyte cultures. 4. Methods for direct identification of fungi from clinical samples. 4.1. DNA isolation. 4.2. Classical PCRbased techniques of direct identification. 4.3. Real-time PCR-based techniques of direct identification. 5. Choice of an optimal method for routine use. 6. Advantages and drawbacks of molecular identification methods applied in mycology. 7. Summary

HUBUNGAN WILAYAH KERJA TERHADAP KEJADIAN JAMUR PADA KARYAWAN PT.PERKEBUNAN NUSANTARA VII

Dermatophytosis was a disease in tissues that contain substances on the horn for examplestratum corneum epidermidis, hair and nails caused by dermatophyte fungi group. Dermatophytosishad keratinofilik nature, each species of dermatophytes had an affinity for a particularhost. Zoophilic dermatophytes which primarily affects animals, and humans. This study aimed todetermine the relationship of the working area on the incidence of fungal infections of the employeesof PT. Plantation Nusantara VII in North Bengkulu. This research was observational analyticcross-sectional design, the independent variable was the area of employment, the dependentvariable was the incidence of fungal infections. The population was 29 employees PT. PerkebunanNusantara VII, samples of all employees PT.Perkebunan Nusantara VII with a total samplingtechnique. Data was analyzed using frequency distribution and bivariate using Chi-Squaretest. The results revealed that the majority of employees working in the processing of fungal infectionthat is equal to 88.2%, which was statistically significant relationship between the workingarea with the incidence of fungal infections. Director PT.Perkebunan Nusantara VII shouldpay more attention to the safety of its employees with a firmer giving rules and providing extensionignore the dangers of the use of personal protective equipment can cause fungal infections.

Infrared spectroscopy as a novel tool to diagnose onychomycosis.

The determination of causative organisms of onychomycosis is still not optimal. There remains a need for a cheap, fast and easy-to-perform diagnostic tool with a high capacity to distinguish between organisms. To determine whether attenuated total-reflectance Fourier transform infrared (ATR-FTIR) spectroscopy can detect and differentiate causative agents in culture-based, ex vivo nail and in vivo nail models. A methodological study was conducted. Both the ex vivo nail model and in vivo pilot study were carried out in an academic university hospital. Analysis of cultured fungi revealed spectral differences for dermatophytes (1692-1606 and 1044-1004 cm-1 ) and nondermatophytes and yeasts (973-937 cm-1 ), confirmed by dendrograms showing an excellent separation between samples from different genera or species. Exploration of dermatophytes, nondermatophytes and yeasts growing on ex vivo nails exposed prominent differences from 1200 to 900 cm-1 . Prediction models resulted in a 96·9% accurate classification of uninfected nails and nails infected with dermatophytes, nondermatophytes and yeasts. Overall correct classification rates of 91·0%, 97·7% and 98·6% were obtained for discrimination between dermatophyte, nondermatophyte and yeast genera or species, respectively. Spectra of in vivo infected and uninfected nails also revealed distinct spectral differences (3000-2811 cm-1 , 1043-950 cm-1 and 1676-1553 cm-1 ), illustrated by two main clusters (uninfected vs. infected) on a dendrogram. Our data suggest that ATR-FTIR spectroscopy may be a promising, fast and accurate method to determine onychomycosis, including identification of the causative organism, bypassing the need for lengthy fungal cultures.

Detection of Trichophyton rubrum and Trichophyton interdigitalein onychomycosis using monoclonal antibodies against Sub6 (Tri r 2).

Onychomycosis is the most prevalent nail disease and is mainly caused by two dermatophyte species Trichophyton rubrum and Trichophyton interdigitale with a frequency in the range of 80% and 20%, respectively. The secreted protease Sub6 of the subtilisin family, which was never detected in vitro growth conditions, was found to be a robust marker of onychomycosis. The aim of this work was to detect tinea unguium using anti-Sub6 monoclonal antibodies in proteins extracted from clinical nail samples. We produced monoclonal antibodies in mice using recombinant Sub6 as an antigen. Selected monoclonal antibodies were tested by Western blot analysis and ELISA on protein extracts from onychomycosis samples. Several monoclonal antibodies used to quantify Sub6 in proteins extracted from clinical nail samples were produced and characterised. We showed that these antibodies were very specific and allowed the detection of T.rubrum and T.interdigitale in onychomycosis. Sub6 was detected in clinical samples infected by T.rubrum and not detected in nails with trauma and other diseases. Anti-Sub6 monoclonal antibodies could be useful for a rapid diagnosis of tinea unguium and/or therapeutic survey of dermatophyte in onychomycosis by ELISA or an immunochromatography device such as a strip test.

Insight into the draft whole-genome sequence of the dermatophyte Arthroderma vanbreuseghemii

Next-generation technologies have prompted efforts towards generating a large repertoire of whole-genome sequences. The dermatophyte Arthroderma vanbreuseghemii has been considered as a good model in which to conduct molecular biological studies on this fungal group. Despite the considerable repertoire of molecular tools developed for this fungus, the lack of genomic data has represented a major limitation, preventing effective implementation of those tools. Herein, the authors report the first draft whole-genome sequence of this dermatophytic species. The size of the draft genome was 23 Mb, exhibiting a GC content of 48.1%. Given the significance of secreted proteases in tissue invasion, a comparative analysis of genes encoding extracellular proteases was performed between A. vanbreuseghemii and other dermatophytes. Furthermore, genes that might be involved in DNA repair also were compared among dermatophytes. Moreover, the complete mitochondrial genome of A. vanbreuseghemii was obtained and shown to consist of 24,287 bp with a GC content of 24%. In conclusion, the availability of genomic data for A. vanbreuseghemii is expected to facilitate the implementation of the molecular tools established for this fungus, enhancing our understanding of the biology of dermatophytes.

Comparison of Two Conventional Methods for Identification of Dermatophyte Fungi

The current study is the identification and isolation dermatophyte species in clinical isolates by both Sabouraud’s Dextrose Agar (SDA) and on Dermatophyte Test Medium (DTM). Clinical specimens of hair, nails and skin scales were collected from patients with dermatophytosis and submitted to direct microscopic examination after immersion in 20% of potassium hydroxide solution. The clinical specimens were cultured on SDA containing chloramphenicol and cycloheximide, and on DTM. Tinea corporis showed the highest prevalent dermatophyte infection among patients (26.7%), followed by Tinea pedis (23.3%), whereas Tinea manuum exhibited the lowest fungal infection (6.7 %). Rural areas revealed the highest prevalence of dermatophyte infection (70.0 %) in comparison to 30.0% in urban areas. Based on the conventional laboratory methods, 30 clinical isolates of dermatophytes showed positive cultures which belong to three genera (Trichophyton, Microsporum and Epidermophyton). Trichophyton mentagrophytes was the most common species (21.7%) isolated among 30 positive dermatophytes, followed by Epidermophyton flocosum (17.4%), then Trichophyton bullosum and Trichophyton tonsurans (13.0%).

Dermatophytes and bacterial superinfectives in tinea pedis patients at Haji Adam Malik Central Hospital, Medan-Indonesia

Background: Tinea pedis is a dermatophyte infection of the feet, especially the toes and soles of the feet. The clinical features of tinea pedis consists of four forms. They are interdigitalis, chronic hyperkeratotic, vesicobullous, acute ulcerative or combination. Tinea pedis mainly interdigitalis form often involved bacterial superinfection, initially Gram-positive bacteria, but Gram-negative bacteria may also be involved. In a state of severe infection, the fungus is getting hard to find, otherwise the bacteria were easier to find, so it is necessary to do fungal and bacterial culture with antibiotic sensitivity tests.Objective: To determine dermatophyte species and bacterial superinfection in tinea pedis at H.Adam Malik General Hospital Medan.Methods: This study is a descriptive observational with cross sectional design. The results of the study are presented in the form of frequency distribution table. These research involved 45 subjects with tinea pedis who met the inclusion and exclusion criteria. We conducted anamnesis, dermatologic examination and collection of samples that were skin scrapings and smears on the lesion. The samples then examined on microscopic directly by using KOH. When KOH result was positive, it followed by fungal and bacterial culture.Results: The number of subjects who suffer from tinea pedis is not much different between men (48.9%) and women (51.1%).The highest prevalence was in age group 37-46 years (28,9%), mostly graduated from elementary school (40%) and mostly housekeepers (22.5%),  with the most prevalence type was interdigitale by 40 cases (88.9%). Dermatophytes T. mentagrophytes were most found by 44.4%, followed by T. rubrum, E. floccosum and T. violaceum. Bacterial superinfection were mostly involved S.aureus by 30,8%, but B. subtilis, S.epidermidis, K.oxytoca, K.pneumoniae, E. coli, P. vulgaris were also found.Conclusion: Dermatophytes T. mentagrophytes were most found followed by T. rubrum. Bacterial superinfection S.aureus were mostly involved.

Therapeutic efficacy of some extracts of hot pepper fruits (Capsicum annuum) and Sumac seeds (Rhus coriaria) on the growth of some isolated dermatophytes species

Background: Fungal infections are one of the common skin diseases with difficulty in their treatment approach. The present efficient drugs for fungal infection are limited.
 Aim: To determine the therapeutic efficacy of plant extracts as alternative antifungal agents.
 Materials and methods: 100 clinical samples [68 from female and 32 from male] were collected during the period from March to July 2017 from subjects attending Dermatology Clinic in Salah Uldean General Hospital. Fungal infection was diagnosed with using KOH wet preparation. Fungal species identified by using conventional approach. The active ingredients existing in the plant extracts were detected and analyzed through qualitative and quantitative detection technique of chemical compounds using a high performance liquid chromatographic device (HPLC). Agar diffusion method was used to determine antifungal activity of plant extracts.
 Results: Direct microscopic examination showed that there were (75%) positive samples, while culture shows (67%) positive samples. The isolated dermatophytes belong to Epidermophyoton, Microsporum, and Trichophyton genus. The predominant dermatophytes were T. rubrum (25%) species. The highest frequency of infection was in the age group of 11-20 years. The sensitivity of the tested fungi to the aqueous and alcoholic plant extracts varies. Alcoholic extract of the hot pepper plant was more effective as antifungal than the aqueous extract of the same plant. However, aqueous hot pepper extracts was more effective against T. mentagrophyte than that of alcoholic extract. Additionally, alcoholic Sumac extract shows higher efficacy that aqueous extract.
 Conclusion: Hot pepper and Sumac extracts show antifungal activity against Microsporum canis, Trichophyton rubrum and T. mentagrophyte.

Unequal distribution of the mating type (MAT) locus idiomorphs in dermatophyte species

The mating type (MAT) locus is the key regulator of sexual reproduction in fungi. In the dermatophytes and other Ascomycetes this genomic region exists in two distinct forms (idiomorphs) and their balanced presence is a precondition for successful mating in heterothallic fungi. But the MAT locus not only drives sexual reproduction, it has also been shown to influence pathogenicity, virulence, and/or morphological changes in pathogenic fungi of the genera Candida, Histoplasma, and Cryptococcus. In order to find out whether there are similar trends in dermatophytes, we investigated the MAT locus of 19 anthropophilic and zoophilic species via Sanger sequencing and primer walking.We identified for the first time the MAT locus idiomorphs of the dermatophyte species Microsporum audouinii (MAT1-2), M. ferrugineum (MAT1-2), Trichophyton schoenleinii (MAT1-2), T. bullosum (MAT1-1), T. quinckeanum (MAT1-1), T. concentricum (MAT1-1), T. eriotrephon (MAT1-1), and T. erinacei (MAT1-2). In addition, we determined the MAT locus sequence for dermatophyte species whose mating type idiomorphs had been described on the basis of results of classical confrontation experiments (e.g. M. canis, MAT1-2) and we confirmed recently published molecular data (e.g. T. rubrum, MAT1-2). Our results corroborate that MAT locus idiomorphs are unequally distributed in the majority of the analyzed species and the ability to mate with a partner of the opposite sex is limited to a few zoophilic species. Clonal spreads are identified that are connected to one of the idiomorphs and a higher virulence and/or a higher transmission rate to humans (T. benhamiae and T. mentagrophytes). For the imbalanced idiomorph distribution pattern we hypothesize that either: (I) one of the mating type idiomorphs may be extinct due to clonal reproduction (e.g., T. rubrum and M. canis), (II) mating partners of one species adapted to different hosts followed by speciation in the new niche (e.g., T. equinum and T. tonsurans) or (III) unisexual reproduction is the next evolutionary stage of propagation in dermatophytes which involves the extinction of one mating idiomorph.

In vitro models of dermatophyte infection to investigate epidermal barrier alterations.

Fungal infections of the skin, known as dermatophytoses, are initiated at the epidermal barrier and lead to dysfunctions of the stratum corneum and cornified skin appendages. Dermatophytosis affects a significant part of the human population and, despite the availability of effective treatments, its prevalence is still increasing. Numerous dermatophyte species are able to induce lesions in both animals and humans, with different clinical pictures and host inflammatory responses. The understanding of the infectious process and of tissue responses has been impeded by discrepancies between observations in vivo or in research models. Indeed, cells cultured as monolayers do not undergo the keratinization process required to study the adherence and invasion of dermatophytes. Animal models lack relevance to study human dermatophytosis because of species-specific differences in the development of lesions and inflammatory responses. This review focuses on the recent development of cultured human skin equivalents, which partly overcomes those limitations and allows improved understanding of the pathogenesis of dermatophytosis in human being, especially the impacts of infection on epidermal barrier integrity.

Exoproteome Analysis of Human Pathogenic Dermatophyte Species and Identification of Immunoreactive Proteins.

Increasing incidence of onychomycosis and tinea pedis in humans of industrialized countries together with deep tissue infections are a therapeutic challenge in clinical mycology. For a better understanding of the pathology and immunology of infection, the authors analyze the exoproteomes of three reference strains of the most common clinical dermatophyte species (Trichophyton rubrum, Trichophyton interdigitale, Arthroderma benhamiae) and of Trichophyton strains isolated from affected patients. Extracellular proteins of those in vitro grown strains are separated via 2D High Performance Electrophoresis and identified by mass spectrometry to find proteins with provoked host immune reactivity. More than 80 secreted proteins including virulence factors such as peptidases and other hydrolases are identified. By Western blotting with respective patient sera, up to 31 proteins with significant antigen-antibody reactions are detected in comparison with control sera, for example, peptidases as well as several oxidoreductases. One protein, beta-glucosidase F2SZI9 seems to be a commonly processed antigen in all Trichophyton infections. These first global exoproteome data of three dermatophyte species can be a stepping stone on the way to further study the molecular mechanisms of Trichophyton pathogenicity-associated traits. Possible candidates for potential new diagnostic methods or vaccination have to be validated in further investigations.

Phenotypic characterization of enzymatic activity of clinical dermatophyte isolates from animals with and without skin lesions and humans.

The pathogenesis of dermatophytoses is associated with the secretion of enzymes degrading the infected tissue components. Although many studies on enzymatic activity of dermatophytes have been conducted over the years, there have been no concrete proposals on the construction of the profile of enzymes characteristic of individual species, genus or ecological types of dermatophytes. The aim of this study was to assess the capability of clinical dermatophyte isolates from both symptomatic and asymptomatic animals and humans to produce different enzymes. Clinical isolates of 234 dermatophyte strains collected during routine examination of animal health were used in this study. The enzymatic production of keratinase, elastase, phospholipase, lipase, protease, DNase and gelatinase as well as the haemolytic activity were evaluated using specific test media. The overall degree of enzymatic activity of the analysed clinical isolates of the dermatophytes was 67%. All tested clinical isolates of different species of dermatophytes showed keratinase activity and 96% additionally exhibited phospholipase activity. The weakest activity among the tested enzymes was demonstrated for elastase and gelatinase. 83% of the isolates of the dermatophytes showed haemolytic activity. Our data indicate that clinical isolates of dermatophytes from different species produce enzymes with different levels of activities. Profile of enzymes characteristic of individual species, genus or ecological types of dermatophytes is possibly dependent upon factors related to the host. The relationship between each enzyme and the occurrence of skin lesions in animals and humans or asymptomatic animal carriers varies on whether the infection is caused by Trichophyton mentagrophytes, Trichophyton verrucosum or Microsporum canis. Interestingly, only keratinase seems to be correlated with the appearance of dermatophyte infections, irrespective of the pathogen species, and elastase is a characteristic enzyme for dermatophyte strains infecting humans. Haemolysis seems to be dependent on host factors and is more common in the case of human dermatophyte isolates.

Microsporum gypseum-infectie bij een paard met erge zomereczeem

Bij een paard met erge zomereczeem en een voorgeschiedenis van corticosteroïdebehandeling voor deze aandoening, werd een welomschreven, cirkelvormig letsel met witte korstvorming opgemerkt. De geofiele schimmel Microsporum gypseum werd geïsoleerd uit het letsel, waarna een vier weken durende topicale behandeling met enilconazole werd gestart. Twee maanden na de behandeling was het letsel volledig hersteld. Deze case-report laat vermoeden dat zomereczeem in combinatie met systemische corticosteroïdebehandeling als risicofactor kan optreden voor een dermatofyteninfectie. Verder wordt het belang van een correcte staalname, isolatie en identificatie van de oorzakelijke kiem bediscussieerd.

Dermatophytosis Caused by Microsporum canis in a Free-Living Maned Wolf (Chrysocyon brachyurus).

Background: Chrysocyon brachyurus is a South American wild canid considered a species near threatened by the International Union for Conservation of Nature and is classified as vulnerable in the National List of Endangered Species. With the increase of the contact between domestic animals, human population and wild animals, there was a greater exposure of the maned wolf to pathogens. Due to the importance of its conservation, the knowledge of emerging infectious diseases that affect this species becomes essential. This report aims to describe the first diagnosed case of dermatophytosis caused by Microsporum canis in a maned wolf.Case: A free-living female maned wolf (Chrysocyon brachyurus), approximately 60 days old, was rescued with presence of alopecia, non-pruritic lesions, one of circular shape located in the nasal plane and the other with signs of scaling and crusts in the region of the left pina. The animal was active, in good general condition and without other significantchanges to clinical examination. Skin scraping was performed for mycological culture of both lesions. Fungal growth on Sabouraud’s agar identified Microsporum canis. Topical therapy with ketoconazole ointment and cleaning of lesions with 0.2% chlorhexidine was instituted. After 20 days of treatment, remission of clinical signs and repilation of affected areas were observed. New mycological cultures of both areas were carried out, which were negative for Microsporum canis.Discussion: Although Microsporum canis is described as causing dermatophytosis in several animal species, it has apparently not yet been reported in maned wolves. Microsporum canis is one of the most isolated zoophilic dermatophytes in domestic cats and is also cited in reports of symptomatic wild felids, such as tigers, in which it has been identified as either the only agent of infection or in association with Trichophyton mentagrophytes. The occurrence is also high in domestic dogs. In the wild canids, Microsporum gypseum has been described in the gray wolf, Trichophyton mentagrophytes in wild foxes, and Microsporum canis in red fox, among others. The finding of zoophilic and geophilic dermatophyte fungi in healthy, free-living and captive wild animals confirms their role as carriers, probable reservoirs and disseminators of these organisms in the environment, and the potential risk they represent as causes of zoonoses. Zoophilic dermatophytes are usually transmitted by contact between individuals and contaminated fomites. In the case of M. canis in the red fox the contact was an asymptomatic domestic cat. In the present case the transmission was not determined, since the animal was rescued from the wild. Regarding the age group, young animals are more susceptible to dermatophytes than adults, probably related to the greater immunity of adults due to previous contacts or even the immaturity of the immune system of the young. The dermatophytosis diagnosis is generally based on anamnesis, physical examination of the lesion, Wood’s lamp examination, microscopic skin scraping, fungal culture, or even histology and PCR tests. In the present case, the final diagnosis was based upon by fungal culture in Sabouraud agar, which allowed to identify the dermatophyte species and,thus, the possible source of infection. Dermatophyte infections can be treated with systemic or topical antifungal medications. Because it was a young animal, it was chosen in the present case only for topical use, which proved to be adequate, probably due to the action of the drug in the areas of alopecia skin and the improvement of the systemic condition of theanimal. Knowing that the maned wolf is considered as a vulnerable species, it is important to know the diseases that affect this species, in order to carry out, when necessary, disease monitoring programs, preventive and therapies, which is essential for its preservation.Keywords: wild animal, disease, skin, dermatophyte.

Specific Identification and Antifungal Susceptibility Pattern of Clinically Important Dermatophyte Species Isolated from Patients with Dermatophytosis in Tehran, Iran

Background: With regard to the increasing number of antifungal-resistant dermatophytes, the requirement for precise identification of causative agents of infections and antifungal susceptibility test is vital. Antifungal susceptibility testing of dermatophytes plays a pivotal role in managing dermatophytosis. The current study aimed at determining antifungal susceptibility profile of 161 important dermatophyte species isolated from Iranian patients. Methods: The current descriptive, cross sectional study was conducted on 508 clinically suspected samples of dermatophytosis collected and identified by conventional methods. All dermatophyte isolates were identified using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The susceptibility of dermatophyte strains to two routine antider-matophyte agents (terbinafine and griseofulvin) was evaluated using micro-dilution method according to CLSI (the clinical and laboratory standards institute) M38-A2 guidelines. Trichophyton rubrum PTCC 5143 and Candida krusei ATCC 6258 were used as quality controls. Results: Among 161 dermatophyte isolates, T. interdigitale was reported as the most frequent species isolated from patients using PCR-RFLP and Microsporum ferruginum was the least isolated species. The minimum inhibitory concentration (MIC) values of griseofulvin and terbinafine were ranged 0.0312-8 and 0.008-4 µg/mL, respectively. The most susceptible and resistant species to griseofulvin were T. interdigitale (MIC = 0.0312 µg/mL) and T. interdigitale/T. rubrum (MIC = 8 µg/mL), respectively. The results indicated that T. verrucosum (MIC = 0.008 µg/mL) was the most susceptible species to terbinafine, whereas T. interdigitale and T. rubrum were the most resistant species to it (MIC = 4 µg/mL). Conclusions: The obtained results assist clinicians to monitor the trend and be able to choose effective medications to treat patients with dermatophytosis, especially in countries such as Iran, where dermatophytosis is still a public health problem. © 2018, Archives of Clinical Infectious Diseases.

Antifungal activity of Lawsonia inermis leaf extract against dermatophytes species

Antifungal activity of Lawsonia inermis leaf extract against dermatophytes species

Assessment of the subtilisin genes in Trichophyton rubrum and Microsporum canis from dermatophytosis

The presence of subtilisin genes (SUBs) coding for serine proteases with activities keratinolytic on dermatophyte species gives them the ability to adhere and degrade keratin during infection process. The aim of this study was to determine the presence of SUBs in Trichophyton rubrum and Microsporum canis from dermatophyte infections. In the current study, dermatophytes were isolated from human and animal with dermatophytosis. Isolates were identified by direct microscopic and laboratory examination; then, the isolates were examined for the presence of the SUB gene family by PCR with specific primers. This study showed the most frequency of SUB gene families (11/17). At least one virulence gene was observed for 94% (16/17) of the isolates. Statistical analysis revealed significant associations between SUB presence and isolates, origin of isolates (animal or human infection) and infection type (skin, hair, and nail) (P < 0.05). This is the first report on the presence/absence of SUB genes in isolates that were obtained from human and animal infection in Iran, which seems to raise the risk to be pathogenic.

Antifungal susceptibility pattern of dermatomycosis in a tertiary care hospital of North India

&lt;p class="abstract"&gt;&lt;strong&gt;Background:&lt;/strong&gt; Dermatomycoses affect the outer layers of the skin, nails and hair without tissue invasion and are often caused by dermatophytic molds, candida &amp;amp; non dermatophytic molds. Although not dangerous, they are important as a public health problem particularly in the immunocompromised. There are limited studies on the efficacy of antifungal agents against dermatophytes in North India.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Methods:&lt;/strong&gt; This study was conducted to test the efficacy of 5 systemic antifungal agents viz. voriconazole, itraconazole, terbinafine, fluconazole &amp;amp; griseofulvin using Microbroth dilution technique.&lt;strong&gt;&lt;/strong&gt;&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Results:&lt;/strong&gt; Three different species of dermatophytes which were isolated from the clinically suspected cases were &lt;em&gt;Trichophyton mentagrophytes&lt;/em&gt;, &lt;em&gt;T. rubrum&lt;/em&gt; and &lt;em&gt;M. gypseum&lt;/em&gt;. According to the obtained results, Itraconazole and Voriconazole showed the lowest MIC range while Fluconazole and Griseofulvin had the highest MIC range for most fungi tested.&lt;/p&gt;&lt;p class="abstract"&gt;&lt;strong&gt;Conclusions:&lt;/strong&gt; Despite several treatment options being available for cutaneous fungal infections, due to an inappropriate response, there is an increasing need for determining an antifungal susceptibility profile for specific fungal strains. This will enable the clinician to select an appropriate antifungal agent with minimal side effects to avoid antifungal resistance and treatment failure.&lt;/p&gt;

Nucleotide Sequence Database Comparison for Routine Dermatophyte Identification by Internal Transcribed Spacer 2 Genetic Region DNA Barcoding.

Conventional dermatophyte identification is based on morphological features. However, recent studies have proposed to use the nucleotide sequences of the rRNA internal transcribed spacer (ITS) region as an identification barcode of all fungi, including dermatophytes. Several nucleotide databases are available to compare sequences and thus identify isolates; however, these databases often contain mislabeled sequences that impair sequence-based identification. We evaluated five of these databases on a clinical isolate panel. We selected 292 clinical dermatophyte strains that were prospectively subjected to an ITS2 nucleotide sequence analysis. Sequences were analyzed against the databases, and the results were compared to clusters obtained via DNA alignment of sequence segments. The DNA tree served as the identification standard throughout the study. According to the ITS2 sequence identification, the majority of strains (255/292) belonged to the genus Trichophyton, mainly T. rubrum complex (n = 184), T. interdigitale (n = 40), T. tonsurans (n = 26), and T. benhamiae (n = 5). Other genera included Microsporum (e.g., M. canis [n = 21], M. audouinii [n = 10], Nannizzia gypsea [n = 3], and Epidermophyton [n = 3]). Species-level identification of T. rubrum complex isolates was an issue. Overall, ITS DNA sequencing is a reliable tool to identify dermatophyte species given that a comprehensive and correctly labeled database is consulted. Since many inaccurate identification results exist in the DNA databases used for this study, reference databases must be verified frequently and amended in line with the current revisions of fungal taxonomy. Before describing a new species or adding a new DNA reference to the available databases, its position in the phylogenetic tree must be verified.

Molecular characterization of dermatophytes isolated from cattle in Plateau State, Nigeria

Dermatophytes from cattle were successfully characterized to species and strain levels for the first time in Nigeria. This study was undertaken to isolate and characterize dermatophytes from cattle in Plateau State, Nigeria. Two molecular techniques were utilized. The first was the use of polymerase chain reaction (PCR) to amplify the internal transcribed spacer regions of the ribosomal DNA using ITS-1 and ITS-4 as primers. This was followed by restriction fragment length polymorphism analysis of the amplified ITS regions using the enzyme MvaI to identify dermatophyte species. The second technique was a PCR using the short oligonucleotide 5′-GACAGACAGACAGACA-3′ as primer for the RAPD typing of the isolates for identification of dermatophytes based on species specific profiles. Profiles of dermatophytes and their correlation with location, site of infection and severity of disease were also investigated. Both PCR-RFLP and RAPD analysis identified 26 Trichophyton verrucosum and 22 Trichophyton mentagrophytes. The PCR-RFLP analysis of the ITS regions produced two distinct profiles for both T. mentagrophytes and T. verrucosum. The first Profile for T. mentagrophytes consisted of two fragments of approximately 320 bp and 280 bp in length while the second was approximately 350 bp and 250 bp in length. The first profile for T. verrucosum consisted of two fragments having bands of approximately 380 bp and 220 bp. The second profile had a single band of undigested fragment of approximately 600 bp in length. Both T. mentagrophytes and T. verrucosum yielded identifiable fragments by RAPD analysis. Six profiles were produced for T. mentagrophytes and the PCR finger prints ranged from 1 to 9 bands with sizes ranging from approximately 350 to 5000 base pairs in size. Amplification of T. verrucosum isolates produced four Profiles. The PCR fingerprints ranged from 5 to 7 bands with sizes ranging from 500 bp–5000 bp. The results indicate that differences in location could contribute to variations in PCR amplicons of dermatophytes and strain differences in dermatophytes may be responsible for variation in clinical dermatophytosis but no significant association was observed between profiles of dermatophytes and the site of infection. The PCR-RFLP analysis of the internal transcribed spacer regions using the primer set ITS1/ITS4 and RAPD analysis using (GACA)4 as primer were successfully used to accurately identify dermatophytes from cattle to species and strain levels. Few molecular studies targeting dermatophytes of cattle are available in the literature. As far as we know, this may be the first report of molecular characterization of cattle dermatophytes from Africa.