The activity of propolis against pathogenic fungi isolated from human infections

Abstract Propolis is a resinous hive product collected by bees from the buds or other parts of plants. It is known for having various biological properties, including antifungal activity. Among the substances present in propolis, flavonoids and phenolic acids and their esters are responsible for its antifungal properties. This means that propolis is ideal for use as an antifungal agent in alternative medicine to treat a number of both topical and systemic infections caused by Candida species and other yeast-like fungi, dermatophyte and nondermatophyte moulds, without the serious side effects typical of synthetic treatment. It is also active against strains of fungi that are resistant to polyenes and azoles, the classes of drugs most commonly used to treat fungal infections. In this article, we review current knowledge about the activity of propolis from different parts of the world and its components in vitro and in vivo against pathogenic fungi isolated from human infections. The article also indicates the possible mechanism of antifungal activity of propolis and its components.

Nannizzia incurvata in Hue city - Viet Nam: Molecular identification and antifungal susceptibility testing.

BACKGROUND: Nannizzia incurvata, a species belonging to the Nannizzia gypsea complex, is considered a neglected pathogen. OBJECTIVE: To detected N. incurvata isolates from dermatophytosis patients in Hue city - Viet Nam, and test the antifungal susceptibility of this species. Moreover, fungal capability to produce hydrolytic enzymes was evaluated. METHODS: Patients' samples were collected and cultured on Sabouraud-chloramphenicol-cycloheximide medium. Dermatophytes isolates were initially macroscopically and microscopically identified. ITS PCR-RFLP and ITS rDNA sequences were performed to determine and confirm species. An ITS Neighbor-Joining phylogenetic tree evaluated the genetic relationship among isolates. Fungal hydrolytic enzymes were examined, including lipase, phospholipase and protease. Antifungal susceptibility testing was carried out by the disk diffusion method. MICs of itraconazole, voriconazole, and terbinafine against these isolates were determined by the broth microdilution method. RESULTS: Twelve isolates of N. gypsea complex were preliminary morphologically identified. PCR-RFLP and ITS-rDNA sequencing identified and confirmed dermatophytes as N. incurvata strains, respectively. An evident polymorphism among isolates was highlighted in the phylogenetic tree. All isolates showed the activity of lipase, phospholipase, and protease production. Overall, all N. incurvata isolates were susceptible to itraconazole, voriconazole, clotrimazole, miconazole, and terbinafine. Few isolates were susceptible to griseofulvin, and none of them were susceptible to fluconazole. CONCLUSIONS: There was a presence of polyclonal N. incurvata isolates in dermatophytosis patients from Hue city, identified by PCR-RLFP and confirmed by ITS sequencing. We confirmed PCR-RLFP as a reliable technique to identify this species. Azole and terbinafine are the optimal choices for N. incurvata treatment except for fluconazole.

Multiplex RT-PCR provides improved diagnosis of skin and nail dermatophyte infections compared to microscopy and culture: a laboratory study and review of the literature.

Dermatophytes are the most common cause of superficial mycosis, estimated to affect 20% to 25% of the general population. We assessed the performance of a novel real-time polymerase chain reaction (RT-PCR) multiplex assay for diagnosis of dermatophytosis. To evaluate sensitivity and specificity, 10 known bacteria and 10 known fungi commonly found on skin, as well as 105 samples with culture confirmed dermatophytosis were tested using Dermatophyte and Fungi assay (AusDiagnostics, Sydney, Australia), a novel multiplex assay for diagnosis of dermatophytosis in skin and nail. This was followed by prospective evaluation of 195 clinical samples for dermatophytosis by both conventional methods and RT-PCR. RT-PCR showed almost two-fold higher sensitivity and high specificity in the diagnosis of skin and nail dermatophytosis compared to traditional microscopy and culture. In addition, RT-PCR demonstrated markedly reduced turnaround time from 4 to 6 weeks to 4 to 6 hours and ability for high throughput.

Matrix-assisted laser desorption/ionisation-time of flight mass spectrometry: Protocol standardisation, comparison and database expansion for faster and reliable identification of dermatophytes.

BACKGROUND: Accurate and early identification of dermatophytes enables prompt antifungal therapy. However, phenotypic and molecular identification methods are time-consuming. MALDI-TOF MS-based identification is rapid, but an optimum protocol is not available. OBJECTIVES: To develop and validate an optimum protein extraction protocol for the efficient and accurate identification of dermatophytes by MALDI-TOF MS. MATERIALS/METHODS: Trichophyton mentagrophytes complex (n = 4), T. rubrum (n = 4) and Microsporum gypseum (n = 4) were used for the optimisation of protein extraction protocols. Thirteen different methods were evaluated. A total of 125 DNA sequence confirmed clinical isolates of dermatophytes were used to create and expand the existing database. The accuracy of the created database was checked by visual inspection of MALDI spectra, MSP dendrogram and composite correlation index matrix analysis. The protocol was validated further using 234 isolates. RESULT: Among 13 protein extraction methods, six correctly identified dermatophytes but with a low log score (≤1.0). The modified extraction protocol developed provided an elevated log score of 1.6. Significant log score difference was observed between the modified protocol and other existing protocols (T. mentagrophytes complex: 1.6 vs. 0.2-1.0, p < .001; T. rubrum: 1.6 vs. 0.4-1.0, p < .001; M. gypseum:1.6 vs. 0.2-1.0, p < .001). Expansion of the database enabled the identification of all 234 isolates (73.5% with log score ≥2.0 and 26.4% with log scores range: 1.75-1.99). The results were comparable to DNA sequence-based identification. CONCLUSION: MALDI-TOF MS with an updated database and efficient protein extraction protocol developed in this study can identify dermatophytes accurately and also reduce the time for identifying them.

Therapeutic efficacy of topically used luliconazole vs. terbinafine 1% creams.

OBJECTIVES: Dermatomycoses of zoophilic origin, especially those caused by Trichophyton mentagrophytes, often pose considerable therapeutic problems. This is reflected in the growing number of strains of this species with resistance to terbinafine caused by a mutation in the squalene epoxidase (SQLE) gene. Therefore, it is reasonable to look for alternative therapies to the commonly used terbinafine. The aim of the present study was to assess the in vivo effectiveness of topical therapy with luliconazole or terbinafine 1% cream. METHODS: Therapeutic efficacy was assessed using direct examination in KOH with DMSO, qPCR analysis with pan-dermatophyte primers and culturing. Moreover, in vitro susceptibility tests for luliconazole and terbinafine were performed. RESULTS: The results demonstrated significantly higher antifungal activity of luliconazole than terbinafine against dermatomycoses caused by T. mentagrophytes. The geometric mean of the MIC value for luliconazole against all T. mentagrophytes strains was 0.002 µg/ml, while this value for terbinafine was 0.004 µg/ml. In all studied cases, 28-day local therapy with luliconazole contributed to complete eradication of the aetiological agent of infection. CONCLUSIONS: Given the increasingly frequent reports of difficult-to-treat dermatophytoses caused by zoophilic terbinafine-resistant strains, the 1% luliconazole cream can be alternative solution in topical therapy.

In vitro antifungal susceptibility patterns of Trichophyton benhamiae complex isolates from diverse origin.

BACKGROUND: Species from the Trichophyton benhamiae complex are mostly zoophilic dermatophytes which cause inflammatory dermatophytosis in animals and humans worldwide. OBJECTIVES: This study was purposed to (a) to identify 169 reference and clinical dermatophyte strains from the T benhamiae complex species by molecular method and adhering to the newest taxonomy in the complex (b) to evaluate the in vitro antifungal susceptibility profile of these strains against eight common and new antifungal agents that may be used for the treatment of dermatophytosis. METHODS: All isolates, mainly originated from Europe but also from Iran, Japan and USA, were subjected to ITS-rDNA sequencing. The in vitro antifungal susceptibility profiles of eight common and new antifungal drugs against the isolates were determined by CLSI M38-A2 protocol and according to microdilution method. RESULTS: Based on the ITS-rDNA sequencing, T benhamiae was the dominant species (n = 102), followed by T europaeum (n = 29), T erinacei (n = 23), T japonicum (n = 10), Trichophyton sp (n = 4) and T eriotrephon (n = 1). MIC ranges across all isolates were as follows: luliconazole: 0.0002-0.002 µg/ml, terbinafine: 0.008-0.125 µg/ml, efinaconazole: 0.008-0.125 µg/ml, ciclopirox olamine: 0.03-0.5 µg/ml, itraconazole: 0.06-2 µg/ml, griseofulvin: 0.25-4 µg/ml, amorolfine hydrochloride: 0.125-4 µg/ml and tavaborole: 1-16 µg/ml. CONCLUSION: Luliconazole, efinaconazole and terbinafine were the most potent antifungals against T benhamiae complex isolates, regardless of the geographic locations where strains were isolated. These data might help dermatologists to develop effective therapies for successful treatment of infections due to T benhamiae complex species.

The prevalence of dermatophytoses in Brazil: a systematic review.

Dermatophytosis is a common cutaneous mycosis worldwide whose prevalence in Brazil is still unknown. This systematic review has estimated the burden of dermatophytoses from updated literature data reported in the general Brazilian population. We used the following databases: Web of Science, Medline/PubMed, Embase, The Cochrane Library and Scopus for studies published between 2011 and 2020. Original articles with an emphasis on prevalence data for dermatophytosis in the Brazilian population, and diagnosed by culture exam or molecular biology were eligible. We also assessed the methodological quality of the studies. A total of 24 articles met the inclusion criteria and were reviewed. The occurrence of dermatophytoses found in the studies ranged from 4-88.50 %. The pooled prevalence of dermatophytosis for the population studies was 25 % (95 % CI: 24.7-25.3 %). The size of the samples used in the studies ranged from 45 to 36 446 participants, and ages ranged up to 98 years old. The populations studied involved mostly women. The presence of tinea unguium (toenail and fingernail) and tinea pedis were the most frequent dermatophytosis, and we observed a predominance of Trichophyton rubrum, T. interdigitale and T. mentagrophytes. The studies were primarily conducted in patient groups with suspected mycoses and were not entirely representative of the general population. Yet we believe that in the future, more collaborative strategies would improve both diagnostic capacity and epidemiological methodologies, associating the prevalence of dermatophytosis with social and environmental risk factors. This review helps to better understand future epidemiological trends in Brazil and the world.

Atypical chlamydoconidium-producing Trichophyton tonsurans strains from Ceará State, Northeast Brazil: investigation of taxonomy by phylogenetic analysis and biofilm susceptibility.

Chlamydoconidium-producing Trichophyton tonsurans strains isolated in Northeastern Brazil have morphological features different from the classic description of this dermatophyte species. This study investigated the phylogenetic relationship of chlamydoconidium-producing T. tonsurans strains isolated in Northeastern Brazil. Also, the effect of terbinafine and farnesol on mature biofilms of T. tonsurans strains was evaluated. The mass spectra of T. tonsurans strains were investigated by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The ITS and LSU loci regions of rDNA and the partial ß-tubulin gene were sequenced and the phylogenetic tree was analysed. The effects of terbinafine and farnesol on mature T. tonsurans biofilms were evaluated through the analysis of metabolic activity, quantification of biomass and observation by scanning electron microscopy. MALDI-TOF MS spectra of the chlamydoconidium-producing T. tonsurans strains differed from the spectrum of the control strain (ATCC 28942), presenting an intense ion peak at m/z 4155 Da. Phylogenetic tree analysis showed that the chlamydoconidium-producing strains isolated in Northeastern Brazil are allocated to a single cluster, differing from strains isolated from other countries. As for mature T. tonsurans biofilms, farnesol reduced biomass and metabolic activity by 64.4 and 65.9 %, respectively, while terbinafine reduced the biomass by 66.5 % and the metabolic activity by 69 %. Atypical morphological characteristics presented by chlamydoconidium-producing T. tonsurans strains result from phenotypic plasticity, possibly for adaptation to environmental stressors. Also, farnesol had inhibitory activity against T. tonsurans biofilms, demonstrating this substance can be explored for development of promising anti-biofilm drugs against dermatophytes.

Detection and identification of dermatophytes based on currently available methods - a comparative study.

AIMS: Accurate identification of dermatophytes is essential for implementing appropriate antifungal treatment and epidemiological analysis. However, the limitations of conventional diagnostics are a frequently discussed topic, and new diagnostic techniques are constantly expanding. In this study, we assess the suitability of conventional diagnostic techniques in comparison to the real-time PCR assay and MALDI-TOF MS in detection and identification of dermatophytes. METHODS AND RESULTS: Strains included in this study were obtained from human and animals with symptomatic, and asymptomatic infection. A direct examination revealed that 31·7 and 60·9% of samples from symptomatic patients, and 25·7 and 60% from asymptomatic animals were positive, as shown by light and fluorescence microscopy respectively. In turn, dermatophytes were isolated from 90·2 and 71·4% of these samples. The pan-dermatophyte primers in real-time PCR assay facilitated detection in 85·3 and 82·9% of the symptomatic and asymptomatic dermatophytoses respectively. Additionally, species-specific PCR assays were positive in 70·7 and 37·1% of these samples. The MALDI-TOF MS analysis yielded positive results consistent with conventional techniques in 97·2 and 72% of symptomatic and asymptomatic infections respectively. CONCLUSIONS: Our study revealed that there is no universal diagnostic method that would be ideal in each of the cases considered. Nonetheless, conventional techniques are still the most effective and reliable tools for mycological diagnostics. SIGNIFICANCE AND IMPACT OF THE STUDY: Dermatologists and veterinarians have difficulties in making a diagnosis of dermatophytoses based only on observed symptoms of fungal infections, as they mimic symptoms of other dermatoses. In this context, a comparative analysis of the results of diagnostics performed using conventional methods and new technologies are crucial for implementing these pioneer methods into routine laboratory practice.

Antifungal susceptibility testing of dermatophytes: Development and evaluation of an optimised broth microdilution method.

BACKGROUND: Dermatophytosis is one of the most common infections affecting 3%-17% of the population. Resistance to antifungals so far was not of concern in the therapeutic management. However, recent reports of terbinafine-resistant strains in several countries are worrisome making antifungal susceptibility testing inevitable. OBJECTIVES: We aimed to develop and evaluate an optimised broth microdilution assay for antifungal drug susceptibility testing of dermatophytes. METHODS: We first studied the effect of different inocula, incubation temperatures and incubation times to establish an optimised assay. Subsequently, we tested 79 clinical strains of 11 dermatophyte species with 13 antifungals. RESULTS: We found inoculating with 0.5-5 × 104 colony forming units (CFU) and incubating at 29°C ± 1°C for 4 days to be appropriate. Terbinafine was the most active antifungal agent with minimum inhibitory concentration (MIC) values ≤ 0.06 µg/mL, expect for one resistant T mentagrophytes strain, which was isolated from an Indian patient. Also, a majority of MICs of other antifungals that are commonly used to treat dermatophytosis were low, except those of fluconazole. Fluconazole MICs do not correlate with the good efficacy in the clinical management. CONCLUSIONS: Our assay enables fast and reliable susceptibility testing of dermatophytes with a large panel of different antifungals. This helps to improve the therapeutic management of dermatophytosis by detecting resistant strains.

Three-gene phylogeny of the genus Arthroderma: Basis for future taxonomic studies.

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, ß-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.

Molecular identification and antifungal susceptibility among clinical isolates of dermatophytes in Shiraz, Iran (2017-2019).

Dermatophytosis is a common superficial mycotic infection affecting individual's quality of life worldwide. The present study aimed to perform species-level identification and evaluate the antifungal susceptibility patterns of dermatophytes isolated in Shiraz, Iran. This cross-sectional study was conducted on clinical samples collected during 2017-2019 from 307 patients suspected of having dermatophytosis. The isolates were identified by direct microscopy, culture and internal transcribed spacer ribosomal DNA sequencing, and their antifungal susceptibility patterns were determined by the microdilution method. Among 307 patients, dermatophytosis was diagnosed by microscopy in 190 (61.8%) subjects and confirmed in 130 (42.3%) cases by both microscopy and culture. It was found out tinea pedis was the most common clinical manifestation, and Trichophyton mentagrophytes was the most prevalent species (28.4%), followed by T tonsurans (23.8%), Microsporum canis (11.5%), T interdigitale (10%), T verrucosum (6.9%), T rubrum (6.9%), T benhamiae (4.6%), T violaceum (3%), T simii (3%), Epidermophyton floccosum (0.7%) and M ferrugineum (0.7%). Moreover, it was revealed that luliconazole with a geometric mean (GM) minimum inhibitory concentration (MIC) of 0.03 µg ml-1 was the most effective agent against all tested isolates. Regardless of species, 30% of isolates responded to high MICs of griseofulvin (MIC90  > 2 µg ml-1 ). The increasing prevalence of nonindigenous species of T simii, T benhamiae and M ferrugineum in Shiraz, Iran, was a notable finding. In addition, infections due to zoophilic species showed an increasing trend. These epidemiological data, along with antifungal susceptibility patterns, may have implications for clinical decision-making and successful treatment.

Species identification of dermatophytes isolated in China by matrix-assisted laser desorption ionisation-time-of-flight mass spectrometry.

BACKGROUND AND OBJECTIVES: Matrix-assisted laser desorption ionisation-time-of-flight mass spectrometry (MALDI-TOF MS) is a novel technique for identifying dermatophytes. This study aimed to detect the limitation of MALDI-TOF MS applied to dermatophytes. METHODS: A total of 113 DNA-sequenced dermatophyte isolates preserved at the Research Center for Medical Mycology of Peking University were selected for this study. Forty-two isolates were selected as reference strains used to create a supplementary database. Seventy-one isolates (Trichophyton rubrum series, T benhamiae series, T mentagrophytes series species and T schoenleinii) were used to evaluate the suitability of the MALDI-TOF MS Biotyper system. MALDI Biotyper 4.0 software was employed to construct the main spectrum profile (MSP) dendrograms. RESULTS: Correct identification rates at the species and genus levels were 90.1% and 91.5%, respectively, using Bruker Filamentous Fungi Library 1.0 combined with the novel database. The MSP dendrogram of the T rubrum series showed unambiguous separation of T rubrum and T violaceum and that of the T benhamiae series distinguished T verrucosum, T benhamiae and T erinacei. Conversely, the MSP dendrogram of the T mentagrophytes series did not successfully distinguish T mentagrophytes, T interdigitale and T tonsurans. CONCLUSION: MALDI-TOF MS showed good performance in the identification and delineation of the T rubrum series and T benhamiae series, but showed poor performance in T mentagrophytes series.

Tinea capitis at the University Hospital of Tizi-Ouzou, Algeria, and first isolation of Trichophyton tonsurans.

Tinea capitis, which are fungal infections caused by some dermatophyte species, are common in developing countries, such as Algeria, where they represent a public health concern. In order to define the epidemiological, clinical and diagnostic features of these infections, a prospective study was conducting from September 2018 to May 2019, at the University Hospital of Tizi-Ouzou (Algeria). PATIENTS: All patients addressed to the Laboratory of Parasitology-Mycology of the University Hospital of Tizi-Ouzou for a suspected Tinea capitis, were included in this study. MATERIALS AND METHODS: Before sampling, contact with animals or soil, presence of similar lesions in the family circle, and previous antifungal or corticosteroid treatment were searched. Mycological examination included direct microscopic examination of the samples, and culture on Sabouraud agar slants at 27°C for up to 4weeks. RESULTS: Out of the 87 samples examined, 46 allowed us to confirm the diagnosis of Tinea capitis, representing a positivity rate of 52.9%. The sex ratio was 1.09 (52.2% males vs. 47.8% females among the infected patients), and the infection mainly involved children of 4-6years (43.3%). Thirty-four strains of dermatophytes were isolated. Microsporum canis was the most frequent species identified (44.1%), followed by Trichophyton mentagrophytes (38.2%). During this study, Trichophyton tonsurans, an unusual dermatophyte species in Algeria, was identified for the first time in our hospital. CONCLUSION: Tinea capitis are still common in Algeria, mainly affecting school-aged children and preschool children. Microsporum canis and T. mentagrophytes are the major causative agents, in agreement with previous studies showing a decrease in frequency of anthropophilic species, in favour of zoophilic species.

In vitro activities of antifungal drugs against a large collection of Trichophyton tonsurans isolated from wrestlers.

BACKGROUND: Trichophyton tonsurans is the most common agent causing tinea gladiatorum in wrestlers, and limited data on susceptibility profiles of Trichophyton tonsurans are available. OBJECTIVES: We aimed to assess the in vitro activity of the common antifungal drug against a large collection of T tonsurans. MATERIALS/METHODS: The in vitro activities to eight common antifungal drugs (sertaconazole, itraconazole, clotrimazole, fluconazole, butenafine, tolnaftate, terbinafine and griseofulvin) against 128 clinical isolates of T tonsurans strains, obtained from wrestlers with dermatophytosis, were performed according to CLSI M38-A2 broth microdilution document. RESULTS: The geometric mean minimum inhibitory concentration was the lowest for tolnaftate (0.022 µg/mL), followed by itraconazole (0.026 µg/mL), terbinafine (0.033 µg/mL), butenafine (0.088 µg/mL), griseofulvin (0.566 µg/mL), sertaconazole (2.875 µg/mL), clotrimazole (3.419 µg/mL) and fluconazole (12.540 µg/mL). CONCLUSIONS: Evaluation of antifungal susceptibility of dermatophytes showed that tolnaftate and itraconazole were the most effective drugs against T tonsurans and fluconazole had the least effect.

Assessing biodiversity shortfalls of freshwater meiofauna from the Atlantic Forest: New species, distribution patterns and the first total-evidence phylogeny of semiplanktonic Gastrotricha.

The Brazilian Atlantic forest is a tropical rainforest recognized as a hotspot of biodiversity, with high species richness and endemicity. This forest extends over a wide latitudinal range, bordering the entire Brazilian coastline, from sea level to high mountains over 2000 m.a.s.L., and presents a variety of environmental conditions and forest physiognomy. Despite many years of intense studies on animal biodiversity in the biome, there is a lack of information on meiofauna taxa causing several shortfalls in biodiversity knowledge of these tiny organisms. In this study, we address some of these shortfalls by describing a new species of Neogossea (Gastrotricha: Chaetonotida) from a lentic ecosystem in southeastern Brazil, surrounded by fragments of Atlantic Forest by using an integrative approach combining different morphological techniques and molecular data. We also point out new hypotheses of homologous structures due to scanning electron microscope observations of the new species. Additionally, we used two numerical methods to assess distribution patterns and historical regionalization of four freshwater meiofaunal taxa (Gastrotricha, Rotifera, Copepoda and Cladocera). For the first time, we accessed the areas of endemism in this biological hotspot based on aquatic fauna with a very peculiar life history. Due to sampling issues and meiofauna species being widespread, our results raise incongruences with previous endemism analyses on vertebrates and arthropods. Finally, we performed the first total-evidence phylogenetic analyses of benthic and semiplanktonic gastrotrichs based on 59 morphological characters and three molecular markers, employing a parsimony approach. The phylogenetic reconstruction supports the hypothesis of a single origin of semiplanktonic gastrotrichs, and both Dasydytidae and Neogosseidae families are monophyletic taxa as well as four non-monotypic genera.

Immunity to commensal skin fungi promotes psoriasiform skin inflammation.

Under steady-state conditions, the immune system is poised to sense and respond to the microbiota. As such, immunity to the microbiota, including T cell responses, is expected to precede any inflammatory trigger. How this pool of preformed microbiota-specific T cells contributes to tissue pathologies remains unclear. Here, using an experimental model of psoriasis, we show that recall responses to commensal skin fungi can significantly aggravate tissue inflammation. Enhanced pathology caused by fungi preexposure depends on Th17 responses and neutrophil extracellular traps and recapitulates features of the transcriptional landscape of human lesional psoriatic skin. Together, our results propose that recall responses directed to skin fungi can directly promote skin inflammation and that exploration of tissue inflammation should be assessed in the context of recall responses to the microbiota.

Trichophyton indotineae sp. nov.: A New Highly Terbinafine-Resistant Anthropophilic Dermatophyte Species.

In this report, we describe the first isolation of two highly terbinafine (TRF)-resistant Trichophyton interdigitale-like strains from a Nepali patient and an Indian patient with tinea corporis in Japan. These strains (designated NUBS19006 and NUBS19007) exhibited a TRF minimal inhibitory concentration (MIC) of > 32 mg/L and contained a missense mutation (Phe397Leu) in squalene epoxidase (SQLE) gene. The internal transcribed spacer (ITS) region sequences amplified from the isolates (NUBS19006 and NUBS19007) were 99.5% identical to Japanese isolates of T. interdigitale and T. interdigitale strain CBS 428.63. The homology of region sequences were also 97.6% identical to T. mentagrophytes strain CBS 318.56. Moreover, the ITS sequences amplified from the isolates were 100% identical to highly TRF-resistant strains of T. interdigitale, which were isolated in Delhi, India, and harbored mutations in SQLE. The urease test on Christensen's urease agar was positive for T. mentagrophytes and T. interdigitale after 7 days of incubation. On the other hand, the type strain of T. rubrum CBS 100081 T and highly TRF-resistant strains (NUBS19006 and NUBS19007) were negative on Christensen urease agar after 7 and 14 days of incubation. Moreover, NUBS19006 and NUBS19007 were also negative reaction on the hair perforation test. To avoid confusion in the taxonomy of the T. mentagrophytes/T. interdigitale complex, we suggest that the highly TRF-resistant Indian strains be considered a new species independent of T. interdigitale, according to clinical and mycological features.

DNA topoisomerase 2 gene polymorphism in dermatophytes.

BACKGROUND: Dermatophytes are a group of keratinophilic fungi of medical importance. Despite a relatively long history of molecular taxonomic studies, there is still a need for information on genetic polymorphism in wider variety of genomic loci. OBJECTIVES: Our goal was to study partial DNA topoisomerase 2 gene (TOP2) polymorphism in dermatophytes. METHODS: We performed DNA sequencing of TOP2 in 26 dermatophyte species along with ribosomal internal transcribed spacer (ITS) sequencing. RESULTS: The number of polymorphic sites in TOP2 data set was similar to that one in ITS data set. Nannizzia species formed paraphyletic group in TOP2 tree. Trichophyton simii was paraphyletic in concatenated TOP2-ITS tree, one of its two clades contained solely Iranian isolates. CONCLUSIONS: Our results revealed several unresolved problems in the taxonomy of dermatophytes, including probable polyphyly of the genus Nannizzia and the species T simii.