Search

Background: With the rapid development of the economy and changes in lifestyle in recent years, the epidemiological pattern of dermatophytosis has changed. We analyzed the epidemiological features of dermatophytosis in a hospital in Hangzhou from 2018 to 2023., Methods: Data from patients with culture-positive dermatophytosis were obtained from the Laboratory Information System of the Mycology Laboratory of our hospital. Dermatophyte isolates were mainly identified by macroscopic and microscopic features of the colonies., Results: There were 32,786 patients with culture-positive dermatophytosis during the 6-year period, including 16,789 male patients and 15,990 female patients. Patients' ages ranged from 1 month to 97 years. Tinea unguium accounted for the largest proportion (14,879, 45.38%), which was mainly prevalent among females (8946, 60.13%). Significant gender differences were observed in all clinical types of dermatophytosis, except for tinea pedis. In total, 15,486 (47.73%) patients aged 21-40 years. The median age of patients with tinea manum and tinea capitis was the highest (49 years) and the lowest (5 years), respectively. Nine species of isolated dermatophytes were identified, of which Trichophyton rubrum accounted for 90.20% (29,573 cases). M. canis accounted for 66.80% of isolated dermatophytes from tinea capitis. The isolation rates of zoophilic species were 43.41% in tinea faciei and 18.90% in tinea corporis., Conclusion: T. rubrum was absolutely predominant agent of dermatophytosis except tinea capitis in which M. canis was the most common organism. The prevalence of zoophilic dermatophytes has increased in tinea faciei and tinea corporis. Tinea unguium was the major clinical form of dermatophytosis, and it was more prevalent among females. The onset peak of dermatophytosis was in the age group of 21-40 years, except for tinea capitis., Competing Interests: Declarations Competing interests The authors have no relevant financial or non-financial interests to disclose. Ethics Approval This is an observational study. The Hangzhou Third People’s Hospital Research Ethics Committee has confirmed that no ethical approval is required. Informed Consent Not applicable., (© 2024. The Author(s).)

Background: Trichophyton indotineae, a newly defined species within the T. mentagrophytes/T. interdigitale complex, has emerged as an epidemiological concern worldwide. However, owing to the limitations of commonly applied fungal identification techniques, T. indotineae remains underreported. In addition, T. indotineae's response to treatment has been described in only a few studies., Objective: To investigate the prevalence, clinical characteristics and treatment outcomes of terbinafine-resistant T. mentagrophytes/T. interdigitale complex infections, as well as to detect T. indotineae cases., Patients and Methods: A retrospective cohort study was conducted on 22 patients with T. mentagrophytes/T. interdigitale complex infections between 2019 and 2023, using either culture or commercial polymerase chain reaction methods. Patient demographics, disease characteristics and treatment responses were recorded. Patients non-responsive to oral terbinafine underwent further analyses, including DNA sequencing of the internal transcribed spacer region for accurate species identification and mutational analysis of the squalene epoxidase (SQLE) gene., Results: The mean age of the patients was 49.7 years (±18.2), with 54.5% men. Terbinafine-resistant T. mentagrophytes/T. interdigitale complex infections were reported in 46.2% of the cohort (n = 6/13 patients; 9 lost to treatment response follow-up), all of whom exhibited extensive dermatophytosis. Among the terbinafine-resistant T. mentagrophytes/T. interdigitale isolates, all five isolates available for fungal analysis were identified as T. indotineae, harbouring SQLE single-point mutations (Phe397Leu and Leu393Ser). Only three of the terbinafine-resistant cases responded to oral itraconazole 200 mg/day, with two responding only to oral voriconazole and one to oral itraconazole 400 mg/day., Conclusion: All cases of T. mentagrophytes/T. interdigitale assessed in this study were identified as T. indotineae, which exhibits SQLE gene mutations. This underscores the importance of integrating methods to detect T. indotineae in routine clinical practice., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Competing Interests: The authors declare no conflict of interest.

Over the past few years, a recalcitrant dermatophytosis has been observed on the Indian subcontinent, including Sri Lanka, which has caused a major public health issue in the region. An emerging species, Trichophyton indotineae, first described as Trichophyton mentagrophytes ITS genotype VIII, is thought to be responsible for this fast-spreading, mostly terbinafine-resistant dermatophytosis. Recalcitrant dermatophytosis is a challenge to dermatologists, and knowing the causative species and antifungal sensitivity in the earlier stage of management would be invaluable. We report a case series of patients with dermatophytosis caused by T. indotineae in Sri Lanka. This is the first detection of this highly terbinafine-resistant strain in Sri Lanka, and existence of this species should be taken seriously by dermatologists and healthcare policymakers for better management of tinea infections and antifungal stewardship in the country.

Trichophyton indotineae is an emerging species of the Trichophyton mentagrophytes complex (TMC), responsible for an epidemic of widespread hairless skin infections that is frequently (50-70%) resistant to terbinafine. In order to initiate appropriate treatment as quickly as possible without waiting for culture positivity (10-15 days) and molecular identification from the strain, we developed a dual quantitative PCR (qPCR) for the direct detection of T. indotineae in clinical samples. We first designed a T. indotineae-specific qPCR assay (TI-qPCR) targeting a single specific polymorphism in the internal transcribed spacer region. Although none of the 94 non-dermatophyte and 7 dermatophyte species were amplified, this TI-qPCR allowed amplification of other TMC species at a lower yield. With equal amounts (0.1 ng) of DNA per reaction, the mean quantitative cycle (Cq) values for T. indotineae and non-indotineae TMC were 27.9 (±0.1) and 38.9 (±0.3), respectively. Therefore, we normalized this assay against a previously validated pan-dermatophyte qPCR assay (PD-qPCR) and relied on the ΔCq [(TI-qPCR) - (PD-qPCR)] to identify T. indotineae versus other TMC species. Dual assay was validated using 86 clinical samples of culture-confirmed T. indotinea and 19 non-indotineae TMC cases. The mean ΔCq for non-indotineae TMC was 9.6 ± 2.7, whereas the ΔCq for T. indotinea was -1.46 ± 2.1 (P < .001). Setting the ΔCq at 4.5 as a cutoff value resulted in 100% specificity for the detection of T. indotineae. This dual qPCR assay quickly detects T. indotineae from skin scrapings, aiding in early diagnosis and treatment for patients with suspected infection., (© The Author(s) 2024. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

Dermatophyte infections (a.k.a. ringworm, tinea) affect an estimated 20%-25% of the world's population. In North America, most dermatophytoses are caused by Trichophyton rubrum or Trichophyton mentagrophytes species complexes. Severe and antifungal-resistant dermatophytoses are a growing global public health problem. A new species of the T. mentagrophytes species complex, Trichophyton indotineae , has recently emerged and is notable for the severe infections it causes, its propensity for antifungal resistance, and its global spread. In this issue of the Journal of Clinical Microbiology , C. F. Cañete-Gibas, J. Mele, H. P. Patterson, et al. (J Clin Microbiol 61:e00562-23, 2023, https://doi.org/10.1128/JCM.00562-23) summarize the results of speciation and AFST performed on North American dermatophyte isolates received at a fungal diagnostic reference laboratory. Within their collection, 18.6% of isolates were resistant to terbinafine (a first-line oral antifungal for dermatophytoses), and similar proportions of T. rubrum and T. indotineae demonstrated terbinafine resistance. The authors also found that T. indotineae has been present in North America since at least 2017. These findings highlight the importance of increased surveillance efforts to monitor trends in severe and antifungal-resistant dermatophytoses and the need for antifungal stewardship efforts, the success of which is contingent upon improving laboratory capacity for dermatophyte speciation and AFST., Competing Interests: The authors declare no conflict of interest.

Trichophyton interdigitale, an anthropophilic species, is one of the main causative agents of tinea unguium and tinea pedis. T. interdigitale and the zoophilic species T. mentagrophytes are morphologically and physiologically very similar. Isolates of the T. interdigitale/T. mentagrophytes complex from around the world have been classified into more than 10 internal transcribed spacer (ITS) genotypes. In this study, we isolated T. interdigitale from Japanese patients and investigated which ITS type was more common. The ITS regions of 29 clinical isolates of T. interdigitale and one clinical isolate of T. mentagrophytes were sequenced. The phylogenetic analysis of the ITS region sequences revealed that the 29 isolates of T. interdigitale belong to ITS type II of T. interdigitale. The one clinical isolate of T. mentagrophytes was in the same cluster with ITS type II* of T. mentagrophytes. One terbinafine-resistant strain of T. interdigitale also belonged to ITS type II of T. interdigitale., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: During the past 5 years, an outbreak of recalcitrant dermatophytosis due to a novel Trichophyton species generally resistant to terbinafine, T. indotineae, has spread out from South Asia to many countries around the World. These isolates cannot be reliably differentiated from other Trichophyton spp. on the basis of morphological traits and the current laboratory diagnostics relies on sequencing of ribosomal DNA ITS region., Objectives: In this study, we aimed to introduce two inexpensive and rapid PCR-based assays for differentiation between T. indotineae and other dermatophytes., Methods: The first introduced assay is based on PCR-restriction fragment length polymorphism (PCR-RFLP) analysis, involving the amplification of TOP2 sequences and the digestion of PCR products by Cfr13I restriction enzyme. The second assay is proposed as conventional endpoint species-specific PCR amplification of the C120-287 intergenic locus. To validate the assays, a total of 191 Trichophyton spp. and 2 Microsporum canis strains with known ITS region sequences were used. From the T. mentagrophytes / T. interdigitale species complex (TMTISC), strains with 18 different ITS genotypes were tested. The sample of TMTISC isolates included 41 T. indotineae strains., Results: TOP2 PCR-RFLP and T. indotineae-specific PCR were positive with testing on DNA of all 41 T. indotineae isolates and two strains of T. mentagrophytes belonging to ITS Types XIII and XVI, but negative with other species and other TMTISC ITS genotypes (n = 152). Therefore, the specificity of both new assays was 99%., Conclusion: The two developed diagnostic assays provide accurate and cost-effective means of identifying cultured T. indotineae isolates., (© 2022 Wiley-VCH GmbH.)

Background: Tinea capitis is a common dermatophyte infection of the scalp primarily affecting children, with less frequent, though not rare, observation in adults., Objective: This study aimed to assess changes in the causative agents of adult tinea capitis over a 60-year period in the Hubei area., Methods: A retrospective, single-centre study was performed on 164 adults with tinea capitis between 1960 and 2020., Results: Out of 1113 cases of tinea capitis, 164 patients were adults, representing 14.7% of all patients. Adult tinea capitis was slightly more prevalent in males (91, 55.5%) than in females (73, 44.5%), but gender difference was not statistically significant between adults and children. Adult tinea capitis was most prevalent between the ages of 18 and 29 years, with a mean age of 22 years. Trichophyton schoenleinii was the most common dermatophyte in adult tinea capitis (78, 47.6%), followed by Trichophyton violaceum (58, 35.4%). Most adult tinea capitis cases before the 1980s were caused by T. schoenleinii, but T. violaceum has become the leading pathogen for recent adult tinea capitis cases., Conclusion: Tinea capitis is not a disease exclusive to children. On the contrary, an upward trend of tinea capitis in adults has been observed in recent years. T. violaceum has become the dominant causative agent of adult tinea capitis in the Hubei area, replacing T. schoenleinii. These results provide a better understanding of the treatment and prevention of tinea capitis in adults., (© 2021 Wiley-VCH GmbH.)

Pathogens from the Trichophyton benhamiae complex are one of the most important causes of animal mycoses with significant zoonotic potential. In light of the recently revised taxonomy of this complex, we retrospectively identified 38 Trichophyton isolates that could not be resolved into any of the existing species. These strains were isolated from Iranian and Czech patients during molecular epidemiological surveys on dermatophytosis and were predominantly associated with highly inflammatory tinea corporis cases, suggesting possible zoonotic etiology. Subsequent phylogenetic (4 markers), population genetic (10 markers), and phenotypic analyses supported recognition of two novel species. The first species, Trichophyton persicum sp. nov., was identified in 36 cases of human dermatophytosis and one case of feline dermatophytosis, mainly in Southern and Western Iran. The second species, Trichophyton spiraliforme sp. nov., is only known from a single case of tinea corporis in a Czech patient who probably contracted the infection from a dog. Although the zoonotic sources of infections summarized in this study are very likely, little is known about the host spectrum of these pathogens. Awareness of these new pathogens among clinicians should refine our knowledge about their poorly explored geographic distribution. IMPORTANCE In this study, we describe two novel agents of dermatophytosis and summarize the clinical manifestation of infections. These new pathogens were discovered thanks to long-term molecular epidemiological studies conducted in Czechia and Iran. Zoonotic origins of the human infections are highly probable, but the animal hosts of these pathogens are poorly known. Further research is needed to refine our knowledge about these new dermatophytes.

Introduction: In Mauritania, only an investigation carried out 26 years ago allows to report epidemiological findings about tinea capitis in Nouakchott. The objective of this study was to reactualize the tinea capitis epidemiological, clinical and mycological profile in school children and to compare this profile in urban and rural area., Patients and Methods: All subscribed students in targeted schools and present during the investigation were examined clinically before samples collection. Mycological diagnosis was made by direct examination and culture in Sabouraud Dextrose Agar with chloramphenicol and supplemented with cycloheximide. The results were considered positive when culture with the presence of dermatophytes was obtained., Results: Prevalence of tinea capitis among school children was 10.50% and it was more important in rural area (12.21%) than in urban area (9.52%). The age group ranged between 6 and 8 years was the most affected (13.58%). According to the gender, male (11.46%) were more affected than female (9.62%). Trichophytic tinea were predominant and Trichophyton soudanense was more isolated followed by Trichophyton rubrum, Trichophyton schoenleinii and Microsporum audouinii., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Trichophyton (T.) mentagrophytes now accounts for an overwhelming majority of clinical cases in India, a new "Indian genotype" (T. mentagrophytes ITS genotype VIII) having been isolated from skin samples obtained from cases across a wide geographical distribution in this country. The conventional diagnostic methods, like fungal culture, are, however, inadequate for diagnosing this agent. Thus, molecular methods of diagnosis are necessary for proper characterization of the causative agent. The shift in the predominant agent of dermatophytosis from T. rubrum to T. mentagrophytes, within a relatively short span of time, is without historic parallel. The apparent ease of transmission of a zoophilic fungus among human hosts can also be explained by means of mycological phenomena, like anthropization.

Arthroderma is the most diverse genus among dermatophytes encompassing species occurring in soil, caves, animal burrows, clinical material and other environments. In this study, we collected ex-type, reference and authentic strains of all currently accepted Arthroderma species and generated sequences of three highly variable loci (ITS rDNA, β-tubulin, and translation elongation factor 1-α). The number of accepted species was expanded to 27. One novel species, A. melbournense (ex-type strain CCF 6162T = CBS 145858T), is described. This species was isolated from toenail dust collected by a podiatrist in Melbourne, during an epidemiological study of four geographical regions of Eastern Australia. Trichophyton terrestre, Chrysosporium magnisporum, and Chrysosporium oceanitis are transferred to Arthroderma. Typification is provided for T. terrestre that is not conspecific with any of the supposed biological species from the former T. terrestre complex, that is, A. insingulare, A. lenticulare and A. quadrifidum. A multi-gene phylogeny and reference sequences provided in this study should serve as a basis for future phylogenetic studies and facilitate species identification in practice., Lay Abstract: The genus Arthroderma encompasses geophilic dermatophyte species that infrequently cause human and animal superficial infections. Reference sequences from three genetic loci were generated for all currently accepted Arthroderma species and phylogeny was constructed. Several taxonomic novelties are introduced. The newly provided data will facilitate species identification and future taxonomic studies., (© The Author(s) 2020. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

Background: Dermatophytoses rank among the most frequent communicable diseases in humans, and the zoonotic transmission is increasing. The zoophilic dermatophyte Trichophyton (T.) benhamiae is nowadays one of the main causes of tinea faciei et corporis in children. However, scientific data on molecular pathomechanisms and specific virulence factors enabling this ubiquitous occurrence are scarce., Objectives: To study tissue invasion and the expression of important virulence factors of T. benhamiae, isolates that were recovered from two groups of hosts (humans vs. guinea pigs (GP)) using an ex vivo skin model., Methods: After confirmation of species identity by ITS sequencing, CFU suspensions of dermatophyte isolates (n = 20) were applied to the skin infection model and cultured. Employing specific immunofluorescence staining techniques, the expression of subtilisin 3 and 6 and metallocarboxypeptidase A was analysed. The general mode of invasion was explored. Results were compared with biopsies of naturally infected GP., Results: All isolates were successfully recovered and proliferated well after application to the infection model. Progressive invasion of hyphae through all skin structures and destruction of explants were observed with early events being comparable to natural infection. An increasing expression of the examined virulence factors towards the end of culture was noticed but no difference between the two groups of isolates., Conclusions: For the first time, important in vivo markers of dermatophytosis were visualised immunohistochemically in an ex vivo skin infection model and in skin biopsies of GP naturally infected with T. benhamiae. More research on the underlying pathomechanisms of dermatophyte infection is urgently needed., (© 2020 The Authors. Mycoses published by Blackwell Verlag GmbH.)

To determine fungal species distribution of interdigital intertrigo among seafarers in Dakar, Senegal, a cross-sectional study was carried out from May to August 2017 among seafarers clinically diagnosed with interdigital intertrigo. A questionnaire was filled to each patient before sampling the affected folds and transporting to Aristide Le Dantec University Hospital where mycological analyses were realized. Species identification by MALDI-TOF MS was performed in Marseille, France. In total, 169 men (21-66 years) were included. Few of them (3%) had a high level of education and the duration of the mycosis exceed 10 years for 88% of patients. Direct microscopic examination (ME) was positive in 34.3%. Among samples with positive ME, 58.6% had positive culture. An overall incidence of 30.2% was found. Patients with confirmed cases aged between 28 and 66 years. Among them, those between 36-50 years were predominant (52.9%). Those with a high level of education were less representative (2%). For 52.1% of patients, the duration of the mycosis was superior to 10 years. Furthermore, 57% of cases were significantly associated with other types of tinea pedis and/or onychomycosis (P=0.03). Culture was positive in 23.7% isolating 43 strains successfully identified at the species level by MALDI-TOF MS for 31 isolates: 20 Candida and 11 dermatophytes. The rest was identified only at the genus level belonged to Fusarium. In definitive, MALDI-TOF MS could be a useful tool for routine and fast identification of dermatophytes, yeasts and NDFF in clinical mycology laboratories., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Background: Dermatophytosis is one of the most frequent superficial mycoses in the world., Main Aim: To describe the cases of skin dermatophytosis and its main aetiologic agents in patients referred to a Mycological Reference Laboratory in Medellín, Colombia., Methods: A retrospective study was carried out with records of patients referred between 1994 and 2016 to the Corporación para Investigaciones Biológicas (CIB), Medellín-Colombia, because of clinical suspicion of skin dermatophytosis., Results: Of a total of 5628 clinical records of patients with suspicion of skin dermatophytosis analysed, 2780 (49.4%) had a proven or probable dermatophytosis diagnosis, 2774 cultures were performed, and aetiologic agents were isolated in 2576 samples (92.9%). The most frequently isolated aetiologic agents were Trichophyton rubrum (44.3%), followed by Trichophyton mentagrophytes complex (33.3%), Epidermophyton floccosum (12.4%), Nannizzia gypseum complex (5.7%, formerly Microsporum gypseum), Microsporum canis (3.5%) and Trichophyton tonsurans (0.8%). The most frequent clinical forms were tinea pedis (72.7%) and tinea corporis (12.7%). In addition, a group of patients (0.9%) developed mixed infections by two dermatophyte agents and another (4.1%) developed infections in more than one anatomical site., Conclusions: The results of the present study are coherent with previous reports where T rubrum and T mentagrophytes complex were the main causative agents of dermatophytosis. However, the increased incidence of N gypsea complex over M canis is worth highlighting., (© 2020 Blackwell Verlag GmbH.)

The most important species of the Trichophyton rubrum group are T. rubrum, causing mainly skin and nail infections, and T. violaceum which is mostly scalp-associated. The status of a third species, T. soudanense, has been under debate. With a polyphasic approach, using molecular phylogenetic techniques, MALDI-TOF mass spectrometry and physiological and morphological analysis, we re-evaluated the T. rubrum complex. Our results support four genetic lineages within the complex each with a distinct morphology and identifiable via MALDI-TOF MS: T. rubrum, T. violaceum, T. soudanense and the T. yaoundei clade. However, ITS and Bt2 sequencing data could not confirm these taxa as four monophyletic species. Our results also suggest that strains formerly identified as T. kuryangei and T. megninii should be considered in future taxonomic studies.

The anthropophilic dermatophyte Trichophyton tonsurans and its zoophilic counterpart T. equinum are phylogenetically closely related. The barcoding marker rDNA internal transcribed spacer (ITS) shows limited variation between these two species. In the current study, we combined molecular approaches with phenotypic data to determine the species boundaries between T. tonsurans (n = 52) and T. equinum (n = 15) strains originating from humans (n = 40), horses (n = 26), and a mouse (n = 1). Culture characteristics and physiology on Trichophyton agar media 1 and 5 were evaluated. Multi-locus sequencing involving ITS, partial large rDNA subunit (LSU), β-tubulin (TUB), 60S ribosomal protein (RPB), and translation elongation factor-3 (TEF3) genes, and the mating-type (MAT) locus was performed. Amplified fragment length polymorphism data were added. None of the test results showed complete mutual correspondence. With the exception of strains from New Zealand, strains of equine origin required niacin for growth, whereas most strains from human origin did not show this dependence. It is concluded that T. tonsurans and T. equinum incompletely diverged from a common lineage relatively recently. MAT1-1 and MAT1-2 are the main distinguishing genes between the two species.

Phylogenetic studies of the family Arthrodermataceae have revealed seven monophyletic dermatophyte clades representing the genera Trichophyton, Epidermophyton, Nannizzia, Lophophyton, Paraphyton, Microsporum, and Arthroderma. Members of the genus Nannizzia are geo- or zoophiles that occasionally infect humans. With the newly proposed taxonomy, the genus Nannizzia comprises thirteen species, i.e., Nannizzia aenigmatica, N. corniculata, N. duboisii, N. fulva, N. graeserae, N. gypsea, N. nana, N. incurvata, N. perplicata, N. persicolor, N. praecox, and two novel species. Nannizzia polymorpha sp. nov. was isolated from a skin lesion of a patient from French Guiana. For the strain originally described as Microsporum racemosum by Borelli in 1965, we proposed Nannizzia lorica nom. nov. The species are fully characterized with five sequenced loci (ITS, LSU, TUB2, RP 60S L1 and TEF3), combined with morphology of the asexual form and physiological features. A key to the species based on phenotypic and physiological characters is provided.

Background: The Trichophyton mentagrophytes complex is the second most common causal agent of dermatophytosis. It comprises five species- T. mentagrophytes , T. interdigitale , T. erinacei , T quinckeanum, and T. benhamie, as well as nine different genotypes of T. mentagrophytes / T. interdigitale -which are morphologically similar; however, their susceptibility to antifungal agents may differ. For targeted therapy and better prognosis, it is important to identify these species at a molecular level. However, since many hospitals lack molecular methods, the actual aetiology of dermatophytosis caused by this complex remains unknown. Objective: To characterize 55 anthropophilic isolates of the T. mentagrophytes complex recovered from a dermatological centre in Yucatán, Mexico. Material and methods: Fifty-five isolates of the T. mentagrophytes complex were obtained from patients with tinea capitis , tinea pedis , tinea corporis , tinea barbae , and tinea unguium . They were characterized by their colonial and microscopic morphology on Sabouraud dextrose agar (SDA) and through the sequencing of a fragment from the region ITS1-5.8S-ITS2. Results: All colonies grown on SDA were white. Forty-six isolates formed colonies with a powdery texture, while nine isolates formed colonies with a velvety texture. The micromorphological features were typical of the T. mentagrophytes complex. The molecular analysis revealed that 55 isolates were microorganisms that belonged to the T. mentagrophytes complex, that 46 formed powdery colonies representing T. mentagrophytes , and that the other nine isolates that formed velvety colonies represented T. interdigitale . The latter nine isolates were obtained from patients with tinea pedis , tinea corporis , and tinea unguium . Conclusions: The colony morphology on SDA led to the identification of 46 isolates as T. mentagrophytes and nine isolates as T. interdigitale . At a molecular level, the species identified by their morphology were identified only as T. mentagrophytes complex., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

There has been a considerable upsurge of extensive, treatment recalcitrant, dermatophytosis presenting as tinea corporis and tinea cruris in India since the past few years. Genome analysis of Trichophytonspecies causing severe superficial dermatophytosis in North India confirmed a unique clade related to the T.mentagrophytes/interdigitale complex, seeming to belong to an early diverging clade of the complex. The Indian Trichophyton species genomes were highly related showing only up to 42 SNPs between any two isolates confirming their clonal origin. Other genetic approaches such as ITS sequencing and multigene phylogeny used in this study were contradictory or inconclusive to show the differentiation of these isolates from T. mentagrophytes/T. interdigitale. Remarkably, high rates of resistance to all three commonly used oral antifungals, i.e., 36% for terbinafine (MICs 4 to ≥32 mg/L), 39.5% for fluconazole (MIC range 32 to ≥64 mg/L) and griseofulvin (Geometric mean MIC ≥ 4 mg/L) were observed. Two important amino acid substitutions (Leu393Phe or Phe397Leu) leading to a terbinafine resistant phenotype were found in the squalene epoxidase protein of all tested terbinafine resistant isolates. All 20 examined genomes presented a high mobility group (HMG) domain transcription factor gene corresponding to mating type (+). Of these, three isolates also showed positivity for both alpha-box and HMG in the genome which might indicate hybridization or an incomplete sexual cycle. Therefore, we highlight the potential of this organism to rapidly spread alleles that might be driving antifungal resistance among its population. This new population of Trichophyton with high rates of in vitro antifungal resistance seems to be driving an ongoing outbreak of dermatophytosis in India. Our study highlights difficulties in identifying isolates from the Trichophyton mentagrophytes/interdigitale clade of the genus using currently available molecular tools. High resistance rates of terbinafine warrant further clinical studies to assess its utility in the treatment of dermatophytosis caused by this clonal strain., (Copyright © 2019 Elsevier Inc. All rights reserved.)

A 6 month-old-female infant from Bahrain visiting Germany with her family for a holiday was seen by us for extensive dermatophytosis of the back, buttocks, chest and groins. Topical treatment by terbinafine for over 2 months was not successful. Other family members including adults and children were treated in Bahrain with topical antifungals and oral voriconazole which was not helpful. Mycological examination performed in Germany revealed the detection of the zoophilic dermatophyte Trichophyton (T.) mentagrophytes. The newly described genotype VIII within the species T. mentagrophytes was identified by sequencing of the "internal transcribed spacer" (ITS) region of the fungal rDNA. This genotype of T. mentagrophytes is the main causative agent of the current epidemic of chronic recalcitrant dermatophytoses in India. Transmission of this Indian genotype of T. mentagrophytes to other countries due to globalization is a serious issue to be considered. Moreover, a significant percentage of these Indian T. mentagrophytes strains are resistant to terbinafine both in vitro and by the way of genetic point mutations in the squalene epoxidase (SQLE) gene. Some are also found to be partially resistant against itraconazole and voriconazole. The point mutation TTC/TTA was found by SQLE mutation analysis in this particular T. mentagrophyte isolate from Bahrain. This point mutation is closely associated with F397L amino acid substitution of the enzyme indicative of in vitro resistance of the dermatophyte against terbinafine. The girl was successfully treated by topical miconazole and later by ciclopirox olamine. This is the first report on an infection due to a terbinafine-resistant T. mentagrophytes strain of the ITS genotype VIII from India in Germany.

The fungi Trichophyton mentagrophytes and T interdigitale account for significant amount of dermatophytosis cases worldwide. These two dermatophytes form a species complex and have a number of ribosomal internal transcribed spacer (ITS) region genotypes, allowing simultaneous species identification and strain typing. Our aim was to describe the geographic distribution of T mentagrophytes/T interdigitale ITS region genotypes and find an association between the genotypes and clinical presentations of respective infections. We performed rDNA ITS region sequencing in 397 Iranian T mentagrophytes/T interdigitale isolates and analysed all available in GenBank entries with sequences of this kind. For the study, 515 clinical annotations were available. Statistical analysis was performed by chi-squared test and Spearman rank correlation analysis. A total of 971 sequences belonged to genotypes with at least 10 geographic annotations and were classified on the basis of exclusive occurrence in a particular region or high relative contribution to a regional sample. We discerned Asian and Oceanian ("KU496915" Type V, "KT192500" Type VIII, "KU315316"), European ("FM986750" Type III, "MF926358" Type III*, "KT285210" Type VI) and cosmopolitan ("FM986691" Type I, "JX122216" Type II, "KP132819" Type II* and "AF170453" Type XXIV) genotypes. There was statistically significant difference in the ITS genotype distribution between different affected body sites. Trichophyton mentagrophytes "KT192500" Type VIII correlated with tinea cruris, T mentagrophytes "KU496915" Type V correlated with tinea corporis, T interdigitale "JX122216" Type II correlated with tinea pedis and onychomycosis. Trichophyton mentagrophytes and T interdigitale genotypes can be associated with distinct geographic locations and particular clinical presentations., (© 2019 Blackwell Verlag GmbH.)

Mating experiments were conducted with four clinical Trichophyton benhamiae isolates, genetically similar to the Trichophyton benhamiae CBS 112371, featuring the plus mating type and with two minus type strains. One minus type strain belonged to the white subgroup, and the other minus type strain, DSM 6916, showed genetic kinship to the yellow subgroup. Only two plus type strains were able to form mature, pigmented gymnothecia with DSM 6916. These two plus type strains demonstrated dark pigmentation and powdery mycelium on Takashio agar, whereas the other three strains exhibited a low degree of pigmentation on the same medium. All five plus strains were able to mate with the minus type strain of their own white subgroup. Cultures from single ascospore isolates showed highly variable morphology and pigmentation. Three genetic markers (ITS, mating type, EF1 alpha) were analyzed in polymerase chain reaction (PCR) experiments with optimized primers and PCR conditions to discriminate between subgroups. Furthermore, RAPD-PCR was used to generate a DSM 6916-specific DNA-fragment which served as an additional genetic marker. Assessing the isolates with recombinant genotypes, it was found that three genetic markers behave like linked genes. The recombination of plus mating type went together with ITS, EF1 alpha and RAPD marker of the DSM 6916 parental strain and was most frequently isolated, whereas plus types recombinants in this case were completely missing. This shows a high imbalance in mating type distribution of recombinants., (© The Author(s) 2019. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

The Trichophyton rubrum species complex comprises commonly encountered dermatophytic fungi with a worldwide distribution. The members of the complex usually have distinct phenotypes in culture and cause different clinical symptoms, despite high genome similarity. In order to better delimit the species within the complex, molecular, phenotypic, and physiological characteristics were combined to reestablish a natural species concept. Three groups, T. rubrum , T. soudanense , and T. violaceum , could be distinguished based on the sequence of the internal transcribed spacer (ITS) ribosomal DNA barcode gene. On average, strains within each group were similar by colony appearance, microscopy, and physiology, but strains between groups showed significant differences. Trichophyton rubrum strains had higher keratinase activity, whereas T. violaceum strains tended to be more lipophilic; however, none of the phenotypic features were diagnostic. The results of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and amplified fragment length polymorphism (AFLP) were partially consistent with the ITS data but failed to distinguish the species unambiguously. Despite their close similarity, T. violaceum , T. soudanense , and T. rubrum can be regarded as independent species with distinct geographical distributions and clinical predilections. Trichophyton soudanense is pheno- and genotypically intermediate between T. rubrum and T. violaceum For routine diagnostics, ITS sequencing is recommended., (Copyright © 2019 Su et al.)

In addition to morphological diagnostics, molecular methods have become available for the diagnosis of suspected dermatophytoses. Since March 2016, INSTAND e. V., in cooperation with the National Reference Laboratory for Dermatophytes, has provided an external quality assessment (EQA) test for the genome detection of dermatophytes twice a year. More than half of the participants used commercial kits for the analysis of the samples. All kits with a high level of accuracy correctly determined the presence of Trichophyton rubrum or no dermatophytes. In species diagnostics beyond Trichophyton rubrum there are large differences between the kits. These are examined in more detail based on clinical studies and the results of the EQA test.

In this study, a novel real-time polymerase chain reaction (PCR) assay (DermaGenius®2.0, PathoNostics BV, Maastricht, The Netherlands) and a recently developed microarray test (EUROArray Dermatomycosis, Euroimmun, Lübeck, Germany) were evaluated regarding their diagnostic specificity to identify dermatophyte DNA. The tests were compared to conventional methods and sequencing. The microarray Dermatomycosis test allows the detection of 50 dermatophytes and definitive identification of 23 dermatophyte species, 6 yeasts and moulds combined in one test. In comparison, real-time PCR is able to identify 11 dermatophytes and one yeast at the species level. Using the EUROArray, 22 out of 24 dermatophyte species were correctly identified. Using real-time PCR, 9 out of the 11 different dermatophytes included in the test kit were correctly identified. Both molecular tests for detection and differentiation of dermatophytes are useful tools for daily clinical practice. The real-time PCR test does not detect as many species, and specificity is slightly lower. However, real-time PCR is a very fast and easy to perform test, especially since no post-PCR step is necessary. Real-time PCR detects the most frequent dermatophytes like T. rubrum, T. interdigitale, and M. canis without any problems. The EUROArray is more elaborate to perform in the lab, due to the hybridization step. However, the EUROArray shows higher specificity and can detect a much broader range of causative agents, including rare species, in dermatomycology.

The fungi Trichophyton mentagrophytes and T. interdigitale are closely related species, causing superficial infections in humans and other mammals. The status of these taxa is a field of long-lasting debates. To clarify their phylogenetic relationships within the genus Trichophyton and sharpen the species boundaries, we performed sequencing of four T. mentagrophytes genomes and also evaluated three previously published multilocus data sets. We performed computational species delimitation analysis on all available in GenBank internal transcribed spacer region (ITS) sequences of Trichophyton spp. Phylogenomic data, phylogenetic network, and species delimitation analyses implied that T. mentagrophytes and T. interdigitale belong to the same phylogenetic species. However, we argue that taxonomic status quo should be retained, from the perspective of epidemiological data and the principle of taxonomic stability. Since there is a correlation between ITS genotype and epidemiological source of an isolate, restriction of T. interdigitale to purely anthropophilic ITS genotypes seems to be reasonable., (© The Author(s) 2018. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

Tinea barbae is a rare dermatomycocis, by definition follicular bound in the beard area of adult men. Manifestation usually starts with erythema accompanied by desquamation. Deeper distribution along terminal hairs leads to folliculitis with formation of pustules and nodes as well as abscesses; fixed adherent yellowish crusts may appear. Frequently there is locoregional swelling of the lymph nodes and occasionally a deterioration of general condition with (sub)febrile temperatures. Often this leads to the initial suspected diagnosis of a bacterial folliculitis barbae or impetigo contagiosa. Tinea barbae is mostly induced by species of the genus Trichophyton (T.). The pathogens are diverse and are mostly zoophilic, sometimes anthropophilic and rarely geophilic dermatophytes. With the help of a specific anamnesis and diagnostic procedure, including mycological examinations, histology and molecular detection of dermatophytes via polymerase chain reaction (PCR), tinea barbae-in our patient induced by T. mentagrophytes-can be rapidly diagnosed. Early initiation and adequate treatment duration lead to restitutio ad integrum.

In a preschool a long-lasting outbreak of Trichophyton (T.) tonsurans tinea capitis was stopped by a concurrent screening of all persons at-risk (N = 264) with the hairbrush technique and a therapy based on clinical picture as well as on the quantitative results of the culture. In addition to the 5 symptomatic patients 10 asymptomatic carriers undetected until now were especially important as vectors. With the rising incidence of T. tonsurans and T. violaceum and the return of Microsporum (M.) audouinii in central Europe such outbreaks are likely to occur more frequently. According to the literature an early and comprehensive screening of the entire at-risk population, a combined antimycotic therapy of the symptomatic and at least a topical therapy of the asymptomatic as well as measures of decontamination are important conditions for successful outbreak management. In our hands the hairbrush technique is a reliable, painless and easy-to-perform screening method which also allows a quantification of fungal load.

Trichophyton schoenleinii is an anthropophilic dermatophyte usually causing tinea favosa. Only few studies have provided data on molecular epidemiology and antifungal profiles of this fungus due to its limited prevalence after 1950s. Forty-nine strains from Asia (n = 27), Africa (n = 10), Europe (n = 10) and from unknown regions (n = 2) were analysed with amplified fragment length polymorphism fingerprinting (AFLP) to reveal intraspecific genetic diversity in this dataset. Amplified fragment length polymorphism fingerprinting genotyping revealed five clusters which did not correspond to geographic origins or clinical characteristics. Additionally, in vitro antifungal susceptibility to seven antifungals was provided for all strains. Terbinafine, ketoconazole, miconazole and itraconazole proved to be the most effective drugs, followed by griseofulvin. No correlation between genotypes and differences in antifungal susceptibility was observed. It is concluded that the AFLP groups are lineages within a single species., (© 2018 Blackwell Verlag GmbH.)

The disease burden of chronic-relapsing and therapy-refractory superficial dermatophytosis dramatically increased in India within the past 5-6 years. In order to evaluate the prevalence of this trend, 201 skin scrapings were collected from patients from all parts of India and were tested for dermatophytes using both fungal culture and a PCR-ELISA directly performed with native skin scrapings. Fungal culture material was identified by genomic Sanger sequencing of the internal transcribed spacer (ITS) region and the translation elongation factor (TEF)-1α gene. In total, 149 (74.13%) out of the 201 samples showed a dermatophyte-positive culture result. Out of this, 138 (92.62%) samples were identified as Trichophyton (T.) mentagrophytes and 11 (7.38%) as Trichophyton rubrum. The PCR-ELISA revealed similar results: 162 out of 201 (80.56%) samples were dermatophyte-positive showing 151 (93.21%) T mentagrophytes- and 11 (6.79%) T rubrum-positive samples. In this study, we show for the first time a dramatic Indian-wide switch from T rubrum to T mentagrophytes. Additionally, sequencing revealed a solely occurring T mentagrophytes "Indian ITS genotype" that might be disseminated Indian-wide due to the widespread abuse of topical clobetasol and other steroid molecules mixed with antifungal and antibacterial agents., (© 2018 Blackwell Verlag GmbH.)

The taxonomy of the dermatophytes has been revisited using a novel multilocus phylogenetic approach, and several neotypes and reference strains have been assigned for Trichophyton species. Single gene sequencing, that is the ITS region, for identification of highly related species T. mentagrophytes/T. interdigitale, warrant reassessment. The sequence database (Westerdijk and NCBI) needs to be updated as several incorrect/obsolete entries of reference and neotype strains of T. interdigitale/T. mentagrophytes hampers correct identification of this complex., (© 2018 Blackwell Verlag GmbH.)

The prevalence of tinea pedis (also known as athlete's foot) in Japanese workers as well as contamination of their footwear by pathogenic filamentous fungi were investigated. Health checks by a dermatologist at a factory located in the Kanto region (Japan) led to a clinical and morphologic diagnosis of tinea pedis in 9 of 19 workers. Scales obtained from the feet and dust obtained from the protective footwear (safety shoes) worn daily in the factory were obtained from these nine subjects and tested using a mycological culture technique. Scales obtained from six of the nine subjects indicated pathogenic filamentous fungi, not only Trichophyton spp., but also Acremonium, which causes symptoms similar to tinea pedis or onychomycosis. Similarly, culture of the dust obtained from the safety shoes yielded pathogenic filamentous fungi in six of the nine subjects, and in four samples Trichophyton spp. was also identified. These findings suggest that cultivable Trichophyton spp. can be detected in approximately 40% of the safety shoes of workers with tinea pedis. The risk of reinfection by pathogenic filamentous fungi is likely increased by wearing dermatophyte-contaminated shoes.

An alarming pan Indian increase in the incidence of superficial dermatophytosis has been noticed over the past 5-6 years. Recent studies have demonstrated emerging predominance of Trichophyton (T.) mentagrophytes as the causative organism in such cases. Interestingly, a distinct Indian genotype of T. mentagrophytes has been identified and recognised with the help of sequencing of the ITS region of the rDNA. That has, however, led to a basic confusion owing to the newly introduced taxonomy of dermatophytes in 2017. According to this most recently suggested classification and new taxonomy of dermatophytes, the former "T. mentagrophytes complex" is differentiated into T. mentagrophytes (zoophilic strains) and T. interdigitale (anthropophilic strains). We have noticed that in some recent studies the causative agent of the chronic, relapsing dermatophytosis outbreak in India has been described as T. interdigitale. In our opinion, it is very likely that these T. interdigitale strains isolated in Delhi and Chennai in India are indeed strains more closely related to the neotype of T. mentagrophytes and not strains of T. interdigitale. We therefore want to underscore the importance of a common nomenclature of species in accordance with the new taxonomy of dermatophytes. This would most likely facilitate better understanding of the issue amongst dermatologists and microbiologists in general. Mistaken identification of Trichophyton isolates not limited to India is very likely to occur due to the lack of appropriate molecular diagnosis which in turn is based on the already published data that presumably wrongly identify one species instead of the other., (© 2018 Blackwell Verlag GmbH.)

Objective: A three-year retrospective study of fungi isolated from samples of patients with suspected fungal skin infections in Eastern Slovakia is presented., Methods: A total of 11,989 samples were collected and investigated with direct microscopic examination using 20% KOH and cultivated in Sabouraud and Mycosel medium. Identification was based on macroscopic and microscopic characteristics., Results: Of the total samples, 61.76% (7,405/11,989) were completely negative and 38.24% positive (4,584/11,989). Dermatophytes accounted for 45.88% of isolates (2,103/4,584), yeasts for 26.79% (1,228/4,584), non-dermatophytes for 15.29% (701/4,584), and Malassezia sp. for 12.4% (552/4,584). Trichophyton rubrum was the most prevalent causative agent (79.08%) implicated in fungal skin infections, followed by Trichophyton interdigitale (10.60%). Less frequent isolates included Trichophyton tonsurans (5.13%), and Trichophyton mentagrophytes (3.14%). Other dermatophytes (Microsporum audouinii, Microsporum gypseum, Microsporum canis, Trichophyton violaceum, Trichophyton verrucosum, and Epidermophyton floccosum) were very rarely identified (each in less than 1% of all samples). The main clinical form of dermatophytosis in the sample was tinea unguium (42.61%), followed by tinea pedis (30.86%), tinea inguinalis (11.65%), tinea corporis (8.04%), and tinea manus (4.76%). Tinea capitis et faciei (2.08%) was more common among children and adolescents., Conclusion: The assessment of data has showed the predominance of tinea unguium among adult patients, tinea capitis et faciei among children, and the prevalent aetiological role of Trichophyton rubrum in fungal skin infections; findings that are in agreement with recent European studies.

Tinea pedis and tinea unguium are common infectious diseases, and many elderly people are reported to contract these infections. In this study, to investigate whether strains of the same origin are spreading inside a long-term care facility, we analyzed Trichophyton rubrum and Trichophyton mentagrophytes, isolated from the residents and staff at the facilities located in the Kanto area, using a genomic analytical method targeting tandem repeat regions in the nontranscribed spacer (NTS) region of ribosomal DNA. Five NTS types were confirmed in T. rubrum. T. rubrum of various types (types 1 to 5) was detected at each facility, but there was no isolate specific to one facility only. Eight NTS types of T. mentagrophytes were detected, and T. mentagrophytes that carried an NTS type that was confirmed at one facility only (types C4II, F4II, and D4II) was isolated. These T. mentagrophytes sequence types were isolated from several subjects residing at the same facility. This study proved that a T. mentagrophytes strain of the same type had spread in long-term care facilities. We believe in the importance of cleaning at a long-term care facility as a countermeasure to the spread of Trichophyton species.

Trichophyton verrucosum is a zoophilic fungus that is the most frequent aetiological agent of dermatophytosis in cattle. During the last few years, the number of cases of T. verrucosum from humans has been increasing constantly, which is correlated with the presence of cattle-rearing farms. We identified and analysed T. verrucosum strains isolated from humans and cattle. Identification was carried out traditionally by correlating both the clinical manifestations with a micro- and macroscopic examination. To confirm the species affiliation fully, molecular differentiation methods were used. Direct analysis revealed the presence of arthrospores. The macro- and micromorphology of the isolates obtained from material sampled was homogeneous, and characterised for T. verrucosum. The phylogenetic analysis based on the ITS sequences demonstrated that the strains formed a monophyletic group with T. verrucosum ATCC10 695 with a support of 99%. The MP-PCR analysis indicates high invariability of genomes of strains from humans and animals. MSP-fingerprinting analysis gives the same results as the MP-PCR analysis. To sum up, the rDNA ITS sequence analysis in combination with macro- and micro-morphology only facilitated T. verrucosum species identification without the possibility of intraspecific differentiation. Finding and testing methods, especially molecular technique, with sufficient discriminatory power, is the present challenge for mycologists., (© 2018 Blackwell Verlag GmbH.)

Dermatophytes are keratinophilic fungi belonging to the family Arthrodermataceae. Despite having a monophyletic origin, its systematics has always been complex and controversial. Sequencing of nuclear ribosomal ITS and D1/D2 rDNA has been proposed as an efficient tool for identifying species in this group of fungi, while multilocus analyses have been used for phylogenetic species recognition. However, the search for new markers, with sequence and size variation, which enable species identification in only one polymerase chain reaction (PCR) step, is very attractive. Inteins seems to fulfill these characteristics. They are self-splicing genetic elements present within housekeeping coding genes, such as PRP8, that codify the most important protein of the spliceosome. The PRP8 intein has been described for Microsporum canis in databases but has not been studied in dermatophytes in any other published work. Thus, our aim was to determine the potential of this intervening element for establishing phylogenetic relationships among dermatophytes and for identifying species. It was found that all studied species have a full-length PRP8 intein with a Homing Endonuclease belonging to the family LAGLIDADG. Phylogenetic analyses were consistent with other previous phylogenies, confirming Epidermophyton floccosum in the same clade of the Arthroderma gypseum complex, Microsporum audouinii close to M. canis, differentiating A. gypseum from Arthroderma incurvatum, and in addition, better defining the Trichophyton interdigitale and Trichophyton rubrum species grouping. Length polymorphism in the HE region enables identification of the most relevant Microsporum species by a simple PCR-electrophoresis assay. Intein PRP8 within dermatophytes is a powerful additional tool for identifying and systematizing dermatophytes.

Trichophyton benhamiae is a zoophilic dermatophyte transmitted to humans mostly from guinea pigs and occasionally other animals. It presents two distinct phenotypes: yellow and white. T. benhamiae was formerly known as Trichophyton species of Arthroderma benhamiae; it was considered part of the T. mentagrophytes species complex, and some authors have incorrectly described the yellow phenotype of T. benhamiae as T. mentagrophytes var. porcellae. Identification of T. benhamiae has been difficult, as it was described under more than three names, two phenotypes, and in several different possible host species. During the past 15 years, human infections due to this dermatophyte have been increasingly reported all over the world. In order to better understand the local epidemiology of T. benhamiae and to compare it to other European countries, we performed a 9-year retrospective study in the Strasbourg University Hospital. We studied 41 dermatophytes (38 isolated from humans and 3 from guinea pigs) identified as T. mentagrophytes var. porcellae or A. benhamiae from January 2008 to December 2016 and verified their identification by ITS (Internal Transcribed Spacer) sequencing. ITS sequencing was performed in 35 of the 41 strains, and they were identified as T. benhamiae (33), T. bullosum (1), and T. eriotrephon (1). The other six remaining strains were identified according to morphology as T. mentagrophytes var. porcellae, name incorrectly used since 2010 for the yellow phenotype of T. benhamiae. ITS sequencing is recommended for accurate identification of this dermatophyte and the culture phenotype (yellow or white) should be specified.

Matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) is becoming a popular technology in clinical microbiology. It is a fast and highly specific method for the routine identification of micro-organisms. In this study, we evaluated the suitability of dermatophyte identification after only 2 days of colony growth using MALDI-TOF MS. Two protein extraction protocols were also evaluated consisting of either formic acid alone or of ethanol-formic acid-acetonitrile to achieve a complete protein extraction. Morphology-based techniques were used as the diagnostic standard methods and MALDI-TOF MS results were obtained using the manufacturer's spectral library. Using the formic acid protein extraction protocol after 2 days of colony growth, 70 and 46% of dermatophytes were properly identified at the genus and species-level respectively. The addition of ethanol-formic acid-acetonitrile extraction protocol increased the identification to 90 and 62%. Based on our observations, we propose a two-step workflow for the fast and reliable identification of dermatophytes after only 2 days of colony growth. This flow chart consists of a first direct deposition procedure with the addition of formic acid, followed by a complete protein extraction when dermatophyte identification is not successful., Significance and Impact of the Study: In this study, a two-step workflow for the identification of clinical dermatophytes using MALDI-TOF analysis and commercially available spectral library was developed. The workflow consists of an initial direct deposition of the sample on the MALDI plate and formic acid protein extraction at 2 days of growth culture; if dermatophyte identification is not successful, a complete protein extraction using ethanol-formic acid-acetonitrile is subsequently performed. Using this workflow, the correct isolate identifications increase up to 90%; of these, 27% are identified at the genus-level, providing sufficient information to start an antifungal treatment. The method here proposed represents a fast and useful approach to differentiate dermatophytes grown in culture., (© 2018 The Society for Applied Microbiology.)

Trichophyton rubrum and Candida species comprise the majority of onychomycosis pathogens. The aim of this study was to evaluate Raman spectroscopy for the differentiation between healthy and either T. rubrum or Candida infected nails. Raman measurements were performed on clippings (N = 52) infected either by T. rubrum (N = 12) or Candida species (N = 14; C. parapsilosis (sensu lato): N = 11, C. glabrata: N = 1, C. albicans: N = 2) with healthy nails (N = 26) used as controls. Systematic spectral differences were observed in the 500-520 cm-1 band region, attributable to a diverting imprint of the disulfide stretching of cystine and cysteine residues among samples. Particularly, Candida infected nails demonstrated a shoulder at 519 cm-1, corresponding to the signal of the less stable gauche-gauche-trans conformation of the disulfide bond. Two additional bands at 619 and 648 cm-1, corresponding to the C-S stretching vibration, were more evident in the T. rubrum infected nails. Finally, a Raman band at 1550 cm-1, attributable to amide II and tryptophan (Trp) content, was undetectable in Candida infected nails. Using principal component analysis (PCA), efficient differentiation of healthy, T. rubrum and Candida species infected nails was achieved. Soft independent modeling of class analogy (SIMCA) and partial least squares-discriminant analysis (PLS-DA) were further applied to generate diagnostic algorithms for the classification of Raman spectra. Both techniques succeeded in modeling clinical nail samples in three groups according to their mycological categories. Raman spectroscopy is a promising method for the differentiation of healthy vs. diseased nails, including efficient differentiation between onychomycosis caused by T. rubrum and Candida species.

The zoophilic dermatophyte Trichophyton verrucosum is the most important causative agent of bovine dermatophytosis. Additionally, it causes profound and poorly healing skin infections in humans indicating the high zoonotic potential. The objective of this study was to establish differentiation of T. verrucosum from other dermatophytes by mass spectrometry and to identify distinct features of the mass spectra. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was successful for identification of this pathogen only after extension of the database of the manufacturer with spectra from T. verrucosum strains, which were identified as such by sequencing of the internal transcribed spacer (ITS) region. MALDI-TOF MS analysis was conducted with 46 field isolates from cattle, two live vaccine strains, and 10 isolates from humans identified as T. verrucosum by sequence analysis of the ITS region. The results suggest a very good agreement of both methods. Comparison with the mass spectra of 68 strains of other keratinophilic fungi revealed that most T. verrucosum wild-type isolates showed a characteristic peak at 7950-7954 m/z, which was missing in the spectra of other keratinophilic fungi and the live vaccine strains. The spectra of T. verrucosum were most similar to the spectra of T. benhamiae, an emerging zoophilic dermatophyte. In summary, MALDI-TOF MS is a powerful and reliable tool to identify T. verrucosum.

In the present study, an innovative top-down liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the identification of clinically relevant fungi is tested using a model set of dermatophyte strains. The methodology characterizes intact proteins derived from Trichophyton species, which are used as parameters of differentiation. To test its resolving power compared to that of traditional Sanger sequencing and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF), 24 strains of closely related dermatophytes, Trichophyton rubrum , T. violaceum , T. tonsurans , T. equinum , and T. interdigitale , were subjected to this new approach. Using MS/MS and different deconvolution algorithms, we identified hundreds of individual proteins, with a subpopulation of these used as strain- or species-specific markers. Three species, i.e., T. rubrum , T. violaceum , and T. interdigitale , were identified correctly down to the species level. Moreover, all isolates associated with these three species were identified correctly down to the strain level. In the T. tonsurans - equinum complex, eight out of 12 strains showed nearly identical proteomes, indicating an unresolved taxonomic conflict already apparent from previous phylogenetic data. In this case, it was determined with high probability that only a single species can be present. Our study successfully demonstrates applicability of the mass spectrometric approach to identify clinically relevant filamentous fungi. Here, we present the first proof-of-principle study employing the mentioned technology to differentiate microbial pathogens. The ability to differentiate fungi at the strain level sets the stage to improve patient outcomes, such as early detection of strains that carry resistance to antifungals., (Copyright © 2018 Dukik et al.)

Dermatophytic granuloma, also called Majocchi's granuloma (MG), is an uncommon infection of the dermis and subcutaneous tissues that can occur in both healthy and immunosuppressed hosts. We present two cases of MG with different clinical features. Both patients had satisfactory relief after treatment with terbinafine.

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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Dermatophyte research has renewed interest because of changing human floras with changing socioeconomic conditions, and because of severe chronic infections in patients with congenital immune disorders. Main taxonomic traits at the generic level have changed considerably, and now fine-tuning at the species level with state-of-the-art technology has become urgent. Research on virulence factors focuses on secreted proteases now has support in genome data. It is speculated that most protease families are used for degrading hard keratin during nitrogen recycling in the environment, while others, such as Sub6 may have emerged as a result of ancestral gene duplication, and are likely to have specific roles during infection. Virulence may differ between mating partners of the same species and concepts of zoo- and anthropophily may require revision in some recently redefined species. Many of these questions benefit from international cooperation and exchange of materials. The aim of the ISHAM Working Group Dermatophytes aims to stimulate and coordinate international networking on these fungi.

Dermatophytes include fungal species that infect humans, as well as those that also infect other animals or only grow in the environment. The dermatophyte species Trichophyton rubrum is a frequent cause of skin infection in immunocompetent individuals. While members of the T. rubrum species complex have been further categorized based on various morphologies, their population structure and ability to undergo sexual reproduction are not well understood. In this study, we analyze a large set of T. rubrum and T. interdigitale isolates to examine mating types, evidence of mating, and genetic variation. We find that nearly all isolates of T. rubrum are of a single mating type, and that incubation with T. rubrum "morphotype" megninii isolates of the other mating type failed to induce sexual development. While the region around the mating type locus is characterized by a higher frequency of SNPs compared to other genomic regions, we find that the population is remarkably clonal, with highly conserved gene content, low levels of variation, and little evidence of recombination. These results support a model of recent transition to asexual growth when this species specialized to growth on human hosts., (Copyright © 2018 Persinoti et al.)

Trichophyton tonsurans, a major pathogen causing tinea capitis and tinea corporis, has been isolated from contact sports-players in Japan. The molecular types of 208 strains isolated between 2011 and 2015 were determined to understand the contemporary Japanese epidemic. Of these, 142 were isolated from practitioners of judo, 28 from wrestlers, 7 from sumo wrestlers, and 31 from individuals with unknown backgrounds. Based on length polymorphisms of the non-transcribed spacer (NTS) region of the ribosomal RNA gene, these 208 strains were divided into 3 subtypes: NTS I (204; 98.1%), II (3; 1.4%), and III (1; 0.5%). Single nucleotide polymorphisms (SNPs) and deletion/insertion profiles in the NTS region, length polymorphisms of the alkaline protease 1 gene, and a SNP in the carboxypeptidase Y gene were identified in 50 NTS I strains isolated between 2011 and 2015, and in 10 strains isolated before 2005. All 60 strains were classified as the same molecular type, with a profile identical to that of type Ib, a major type in the United States of America. These results indicate that NTS I strains isolated in Japan are clonal and independent of the type of sports activity.

Background: Formerly only referred to as a subspecies (T. mentagrophytes var. quinckeanum), T. quinckeanum once again constitutes a distinct species according to the updated taxonomy of dermatophytes., Patients and Methods: During routine diagnostic tests conducted at the Mycology Laboratory, Mölbis, Germany, between 11/2013 to 1/2017 (three years and three months), all specimens sent in were examined for T. quinckeanum. Molecular biology methods employed included: 1) DNA hybridization (PCR ELISA), 2) gene sequencing of the ITS region and TEF-1α, and 3) in some cases, MALDI-TOF mass spectrometry., Results: Overall, 62 strains of T. quinckeanum were found. Sixty-eight percent of patients were female; 43 % were children and adolescents (≤ 19 years of age). Cats were a frequent source of infection. Sequencing of all 62 strains revealed a concordance of 100 % with T. quinckeanum sequences contained in the NCBI database. Isolates analyzed by MALDI-TOF mass spectrometry showed specific spectra., Conclusions: In Germany, the zoophilic dermatophyte T. quinckeanum currently appears to be more prevalent than expected. T. quinckeanum strains were isolated from children and adults with dermatomycosis and tinea capitis. Sources of infection with T. quinckeanum include small rodents (mice), horses, and - remarkably commonly -  cats. Given that unequivocal morphological identification of this dermatophyte is not always possible, molecular methods have to be employed in the diagnosis., (© 2017 Deutsche Dermatologische Gesellschaft (DDG). Published by John Wiley & Sons Ltd.)