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Although the fungal microbiome in human skin mainly comprises lipophilic yeasts, including Malassezia species, these microorganisms can cause various dermatitis conditions, including pityriasis versicolor, seborrheic dermatitis, folliculitis, and atopic dermatitis, depending on the host condition. Both Malassezia globosa and Malassezia restricta are major species implicated in Malassezia-related dermatitis. However, the pathogenicity of these microorganisms has not been revealed at the genetic level owing to the lack of a genetic recombination system. Therefore, we developed a gene recombination system for M. globosa using Agrobacterium tumefaciens-mediated gene transfer of the target gene FKB1, which encodes the FKBP12 protein that binds the calcineurin inhibitor tacrolimus. The wild-type strain of M. globosa was sensitive to tacrolimus, whereas the FKB1 deletion mutant was resistant to tacrolimus. Reintroduction of FKB1 into the FKB1 deletion mutant restored wild-type levels of susceptibility to tacrolimus. Moreover, an FKB1-eGFP fusion gene was generated and expression of this fusion protein was observed in the cytoplasm. This newly developed gene recombination system for M. globosa will help further our understanding of the pathogenesis of M. globosa-related dermatitis at the genetic level.

Malassezia restricta is the most predominant fungus in the microbiome of human skin. This microorganism can cause or exacerbate Malassezia-associated skin dermatitis, seborrheic dermatitis, atopic dermatitis, and pityriasis versicolor. The virulence factors of M. restricta have not been analyzed because a gene recombination system has not been developed. In this study, we established an Agrobacterium tumefaciens-mediated gene transfer (ATMT) system, optimized for generating gene-deficient mutants of M. restricta. A mutant of FKB1 gene, which encodes the FKBP12 protein that binds to the calcineurin inhibitor tacrolimus, was generated using the ATMT system. Subsequently, the FKB1 gene was reintroduced into the FKB1 gene-deficient mutant for obtaining a gene-complemented strain. The wild-type strain of M. restricta was sensitive to tacrolimus, whereas the FKB1 gene-deficient mutant was resistant to tacrolimus; the phenotypic drug susceptibility in the mutant was restored by reintroducing the FKB1 gene. Contrastingly, the FKB1 gene-deficient mutant was not resistant to cyclosporine A, which also inhibits calcineurin by binding to cyclophilin A. The gene recombination system for M. restricta will facilitate in elucidating the molecular mechanisms causing Malassezia-associated dermatitis.

We analyzed the short sequence repeats (SSRs) of the intergenic spacer (IGS) region 1 of the ribosomal RNA genes in Malassezia globosa and Malassezia restricta, which predominantly colonize the scalp in androgenetic alopecia (AGA). No AGA-specific SSRs were found in the M. globosa IGS region, whereas a (CT)

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objects Lipophilic Malassezia species are abundant in the scalp microbiome; we investigated the scalp microbiome and sebum composition of patients with androgenic alopecia (AGA) and aimed to identify factors accelerating AGA progression. Materials and Methods Scalp scale samples (swabs) were collected from 55 male Japanese patients with AGA and 63 healthy individuals. Fungal rRNA genes were amplified by PCR and the amplicons were sequenced on the MiSeq platform. The extent of fungal colonization was determined by qPCR. We used gas chromatography/mass spectrometry to measure the sebum levels of free fatty acids, diglycerides, triglycerides, squalene, free cholesterol, cholesterol esters, and wax. Results and Discussion Malassezia restricta predominated in all AGA (64.7%) and non-AGA age groups (44.6%). qPCR revealed that Malassezia colonization was more extensive in the AGA than non-AGA group, regardless of age; the Malassezia level was significantly higher in ADA subjects aged 50–59 than 30-49 years. The TG level was significantly higher in the AGA than non-AGA group (P Conclusion Thus, the scalp fungal microbiome and sebum composition may influence AGA development.

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM Objects Malassezia restricta and M. globosa cause or exacerbate Malassezia -associated skin, seborrheic, and atopic dermatitis, as well as pityriasis versicolor, but the virulence factors remain unclear because between-species genetic recombination has not yet been achieved. We built an Agrobacterium tumefaciens-mediated gene transfer (ATMT) system that generated gene-knockout mutants of both Malassezia species. Materials and Methods The binary vector pAg1-Δfkb1::NAT1 was introduced into M. restricta CBS 7877 and M. globosa CBS 7966 via ATMT; FKB1 was replaced. Then, FKB1 was re-introduced into the mutants to counteract the deficiencies. Results and Discussion The medium acetosyringone concentration and temperature, as well as the co-cultivation ratios of A. tumefaciens and the Malassezia strains, affected recombination efficacy. We generated a mutant of the FKB1 gene (which encodes the FKBP12 protein that binds to the calcineurin inhibitor tacrolimus). Wild-type M. restricta and M. globosa were tacrolimus-sensitive, while the FKB1 mutants were tacrolimus-resistant; drug susceptibility was restored by reintroducing FKB1. Conclusion Studies on recombination between M. restricta and M. globosa will aid elucidation of the molecular mechanisms underlying Malassezia -associated dermatitis.

Four new yeast species belonging to the genus Apiotrichum and two new yeast species belonging to Cutaneotrichosporon are described for strains isolated from guano samples from bat-inhabited caves in Japan. In 2005, we reported these isolates as Trichosporon species based on sequence analyses of the D1/D2 domain of large subunit (LSU) rRNA genes according to available basidiomycetous yeast classification criteria; however, to date, they have not been officially published as new species with descriptions. Their phylogenetic positions have been reanalysed based on comparison of internal transcribed spacer (ITS) region sequences (including the 5.8S rRNA gene) and the D1/D2 domain of the LSU rRNA gene with those of known species; we confirmed clear separation from previously described species. Physiological and biochemical properties of the isolates also suggest their distinctiveness. Therefore, we describe Apiotrichum akiyoshidainum (holotype JCM 12595T), Apiotrichum chiropterorum (JCM 12594T), Apiotrichum coprophilum (JCM 12596T), Apiotrichum otae (JCM 12593T), Cutaneotrichosporon cavernicola (JCM 12590T) and Cutaneotrichosporon middelhovenii (JCM 12592T) as new species. C. cavernicola showed particularly distinctive morphology including large inflated anomalous cells on the hyphae and germination from the cells, although clear clamp connections on the hyphae were not confirmed. Further study is needed to elucidate the morph of this species.

Trichophyton tonsurans is a major causative fungus of human dermatophytosis, which has been isolated from contact sport players in Japan. The microbiome in the scalp of judoists with or without T. tonsurans infection was analyzed to investigate the correlation between T. tonsurans infection and microbiome profile. Among 30 members of the same judo team in a high school, samples were collected by scrubbing their scalp with shampoo hairbrushes; then, DNA was extracted directly from the obtained scales. Twenty-seven datasets were subjects for microbiome analysis and T. tonsurans was detected in six members (no T. tonsurans-positive participants had scalp lesions). Regarding the fungal microbiome, Cyphellophora were more abundant in the T. tonsurans-positive group (TP) than T. tonsurans-negative group (TN) (P < 0.05). Regarding the Malassezia microbiome, Malassezia caprae were more abundant in TP than TN (P < 0.01). Regarding the bacterial microbiome, Lactococcus, Actinobacillus, Beijerinckiaceae and Xanthomonas were more abundant in TP than TN (P < 0.05). Also, the Shannon diversity index revealed no significant diversity between TP and TN, and 3-D principal coordinate analysis revealed no clear separation between TP and TN. There was practically no difference in microbiome between TP and TN, indicating that T. tonsurans could colonize humans regardless of their original microbiome. T. tonsurans coexisted with other fungi and bacteria without affecting species diversity in asymptomatic carriers. To our knowledge, this is the first investigation of the correlation between T. tonsurans infection and microbiome profile.

Modified Leeming and Notman agar medium (mLNA) has been widely utilized to grow lipophilic fungi belonging to the genus Malassezia. We developed a new artificial sebum-containing mLNA to obtain higher yields of Malassezia spp. The olive oil in mLNA was replaced with artificial sebum composed of triglyceride (triolein), diglyceride (glyceryl distearate), fatty acids (palmitic acid, myristic acid, pentadecanoic acid, and oleic acid), and squalene, and the Tween 60 was replaced with glyceryl stearate. Nine human-associated Malassezia species grew well on the artificial sebum-containing mLNA, and the yield of the most predominant fungus on human skin, Malassezia restricta, in artificial sebum-containing mLNA was double that in standard mLNA.

The skin microbiome and sebum may be associated with inflammation-related diseases of the scalp. To assess the pathogenesis and progression of androgenetic alopecia (AGA), we analyzed the composition of sebum and the bacterial and fungal microbiomes of the scalps of 118 Japanese male individuals with and without AGA, then discussed their roles in the pathogenesis of AGA. Sebum triglyceride and palmitic acid contents were higher in the AGA group than in the non-AGA group. Malassezia restricta, a lipophilic fungus that consumes palmitic acid, was abundant on the scalps of patients with AGA. Cutibacterium, Corynebacterium, and Staphylococcus were the most common genera in both groups, and patients with AGA exhibited scalp dysbiosis (increased abundance of Cutibacterium and decreased abundance of Corynebacterium). Our findings suggest that both sebum and the bacterial and fungal microbiomes of the scalp may be involved in the development of AGA.

The proportion ofStaphylococcus aureusin the skin microbiome is associated with the severity of inflammation in the skin disease atopic dermatitis.Staphylococcus epidermidis, a commensal skin bacterium, inhibits the growth ofS.aureusin the skin. Therefore, the balance betweenS.epidermidisandS.aureusin the skin microbiome is important for maintaining healthy skin. In the present study, we demonstrated that the heat-treated culture supernatant ofDelftia acidovorans, a member of the skin microbiome, inhibits the growth ofS.epidermidis, but not that ofS.aureus. Comprehensive gene expression analysis by RNA sequencing revealed that culture supernatant ofD.acidovoransincreased the expression of genes related to glycolysis and the tricarboxylic acid cycle (TCA) cycle inS.epidermidis. Malonate, an inhibitor of succinate dehydrogenase in the TCA cycle, suppressed the inhibitory effect of the heat-treated culture supernatant ofD.acidovoranson the growth ofS.epidermidis. Reactive oxygen species production inS.epidermidiswas induced by the heat-treated culture supernatant ofD.acidovoransand suppressed by malonate. Further, the inhibitory effect of the heat-treated culture supernatant ofD.acidovoranson the growth ofS.epidermidiswas suppressed byN-acetyl-L-cysteine, a free radical scavenger. These findings suggest that heat-resistant substances secreted byD.acidovoransinhibit the growth ofS.epidermidisby inducing the production of reactive oxygen species via the TCA cycle.

Citrobacter koseri, an aerobic Gram-negative bacterium, is isolated from the human skin and intestinal tract. Here, we report the complete genome sequence of Citrobacter koseri strain MPUCK001, which has a 4.9-Mbp genome, containing 4,536 protein-coding sequences.

Four new yeast species belonging to the genus

Staphylococcus aureus is a resident skin bacterium involved in the exacerbation of atopic dermatitis. Here we report that S. aureus regulates the tricarboxylic acid (TCA) cycle via the production of pyruvate for tolerance to betamethasone valerate (BV), an anti-inflammatory drug used in the treatment of atopic dermatitis. The addition of BV or clobetasol propionate to the medium among 5 different anti-inflammatory steroids delayed the growth of S. aureus. Comprehensive gene expression analysis by RNA-seq revealed that BV increased the expression of genes related to glycolysis in S. aureus. Pyruvate, a product of glycolysis, suppressed the S. aureus growth inhibition by BV. The addition of oxaloacetate, a compound in the TCA cycle biosynthesized from pyruvate, was also suppressed the inhibitory effect of BV. Malonate, an inhibitor of succinate dehydrogenase in the TCA cycle, increased the inhibitory effect of BV on the growth of S. aureus. These findings suggest that S. aureus promotes tolerance to BV, an anti-inflammatory steroid, by regulating the TCA cycle via the production of pyruvate.

The aim of this study was to develop a method for the rapid detection of Gardnerella vaginalis, which is proposed to play a key role in the pathogenesis of bacterial vaginosis. Specific loop-mediated isothermal amplification (LAMP) primers were designed and used to detect target DNA within 45 min under isothermal conditions. Comparative screening indicated that the LAMP assay is superior to PCR in terms of rapidity, and is equivalent in sensitivity and specificity. This LAMP assay can be used for rapid screening and detection of G. vaginalis in vaginal samples; the limit of detection is 10 fg DNA.

Vulvovaginal candidiasis (VVC) is a common superficial infection of the vaginal mucous membranes caused by the fungus Candida albicans. The aim of this study was to assess the mechanisms underlying the inhibitory effects of the culture supernatants of Lactobacillus gasseri and L. crispatus, the predominant microbiota in Asian healthy women, on C. albicans biofilm formation. The inhibition of C. albicans adhesion to HeLa cells by Lactobacillus culture supernatant was also investigated. Candida albicans biofilm was formed on polystyrene flat-bottomed 96-well plates, and the inhibitory effects on the initial colonization and maturation phases were determined using the XTT reduction assay. The expression levels of biofilm formation-associated genes (HWP1, ECE1, ALS3, BCR1, EFG1, TEC1, and CPH1) were determined by reverse transcription quantitative polymerase chain reaction. The inhibition of C. albicans adhesion to HeLa cells by Lactobacillus culture supernatant was evaluated by enumerating viable C. albicans cells. The culture supernatants of both Lactobacillus species inhibited the initial colonization and maturation of C. albicans biofilm. The expression levels of all biofilm formation-related genes were downregulated in the presence of Lactobacillus culture supernatant. The culture supernatant also inhibited C. albicans adhesion to HeLa cells. The culture supernatants of L. gasseri and L. crispatus inhibited C. albicans biofilm formation by downregulating biofilm formation-related genes and C. albicans adhesion to HeLa cells. These findings support the notion that Lactobacillus metabolites may be useful alternatives to antifungal drugs for the management of VVC.

A rapid detection method for Lactobacillus crispatus and Lactobacillus iners , which are important for maintaining a healthy vaginal environment, was developed using loop-mediated isothermal amplification (LAMP). The LAMP assay had a lower limit of detection of 10 fg DNA and could detect both species within 45 min.

Malodorants in the human axilla are produced from human biogenic precursors by axillary bacterial enzymes. In the present study, we used pyrosequencing analysis to identify the axillary bacterial microbiota of 13 Japanese male subjects with cumin-like, spicy body odor (C type), and 9 with milky, skin-based body odor (M type). Anaerococcus, Corynebacterium, and Staphylococcus predominated in both C- and M-type subjects, followed by Moraxella and Peptoniphilus. These genera accounted for 96.2-99.9% of the total bacterial population, except in the microbiota of one C-type subject. However, the axillary bacteria in C-type subjects were more abundant than that in M-type subjects. These results suggest that the level of colonization by axillary bacteria is important for the production of malodorants.

Strict microbial control is required in pharmaceutical manufacturing facilities, for which environmental microbial monitoring is fundamental. Appropriate microbial control is based on understanding the abundance and community structure of the microbes in the target environment, but most microbes are not culturable by conventional methods. Here, we determined the bacterial abundance and assessed the environmental microbiome in a pharmaceutical manufacturing facility using rRNA gene-targeted quantitative PCR (qPCR) and high-throughput sequencing of rRNA gene fragments. A commercially available microbial particle counter was also used for real-time measurements. In the air of the first gowning room and the passageway of the facility, the microbial particle number determined by both the particle counter and qPCR was ca. 104/m3; the number of microbial particles was about 100 times the number of culturable bacteria. Thus, the measurement of microbes using the particle counter was accurate. In the second gowning room of the facility, managed by a HEPA filter, the number of particles in the air was dependent on human movement, and was below the detection limit around 10 min after movement. Bacteria of the phyla Proteobacteria, Firmicutes, and Actinobacteria were frequently detected in samples from the facility; these bacteria are constituents of the human microbiota. Among fungi, Aspergillus and Cladosporium were detected in the air, and Malassezia was dominant on the walls. Our results provide fundamental data for the evaluation and control of microbes in pharmaceutical and food industry facilities.

We previously reported that the Malassezia microbiota of the external auditory canal were composed principally of Malassezia slooffiae and Malassezia restricta. In the present study, the fungal microbiota of the external auditory canal were comprehensively analyzed by pyrosequencing on an FLX-454 platform. Of 17 samples tested, the fungal microbiota of 14 samples were composed of > 90% Malassezia species; either M. slooffiae or M. restricta was abundant in individual samples. The external auditory canal exhibits a lower fungal diversity than do other bodily sites.

Seborrheic dermatitis (SD) is a chronic inflammatory dermatologic condition in which erythema and itching develop on areas of the body with sebaceous glands, such as the scalp, face and chest. The inflammation is evoked directly by oleic acid, which is hydrolyzed from sebum by lipases secreted by skin microorganisms. Although the skin fungal genus, Malassezia, is thought to be the causative agent of SD, analysis of the bacterial microbiota of skin samples of patients with SD is necessary to clarify any association with Malassezia because the skin microbiota comprises diverse bacterial and fungal genera. In the present study, bacterial microbiotas were analyzed at non-lesional and lesional sites of 24 patients with SD by pyrosequencing and qPCR. Principal coordinate analysis revealed clear separation between the microbiota of non-lesional and lesional sites. Acinetobacter, Corynebacterium, Staphylococcus, Streptococcus and Propionibacterium were abundant at both sites. Propionibacterium was abundant at non-lesional sites, whereas Acinetobacter, Staphylococcus and Streptococcus predominated at lesional sites; however, the extent of Propionibacterium colonization did not differ significantly between lesional and non-lesional sites according to qPCR. Given that these abundant bacteria hydrolyze sebum, they may also contribute to SD development. To the best of our knowledge, this is the first comprehensive analysis of the bacterial microbiotas of the skin of SD patients.

As DNA sequences of the intergenic spacer (IGS) region in the rRNA gene show remarkable intraspecies diversity compared with the small subunit, large subunit, and internal transcribed spacer region, the IGS region has been used as an epidemiological tool in studies on Malassezia globosa and M. restricta, which are responsible for the exacerbation of atopic dermatitis (AD) and seborrheic dermatitis (SD). However, the IGS regions of M. sympodialis and M. dermatis obtained from the skin of patients with AD and SD, as well as healthy subjects, lacked sequence diversity. Of the 105 M. sympodialis strains and the 40 M. dermatis strains, the sequences of 103 (98.1 %) and 39 (97.5 %), respectively, were identical. Thus, given the lack of intraspecies diversity in the IGS regions of M. sympodialis and M. dermatis, studies of the diversity of these species should be performed using appropriate genes and not the IGS.

Background Euphorbia supina (ES) plant has been used as treatment for inflammatory conditions. The antibacterial effect and the anti-inflammatory mechanism of ES for Propionibacterium (P.) acnes-induced inflammation in THP-1 cells and acne animal model remain unclear. Therefore, the objective of the present study was to determine the antibacterial and anti-inflammatory activities of ES against P. acnes, the etiologic agent of skin inflammation. Method The antibacterial activities of ES were tested with disc diffusion and broth dilution methods. Cytotoxicity of ES at different doses was evaluated by the MTT assay. THP-1 cells were stimulated by heat-killed P. acnes in the presence of ES. The pro-inflammatory cytokines and mRNA levels were measured by ELISA and real-time-PCR. MAPK expression was analyzed by Western blot. The living P. acnes was intradermally injected into the ear of BLBC/c mice. Subsequently, chemical composition of ES was analyzed by liquids chromatography-mass spectrometry (LC-MS). Result ES had stronger antibacterial activity against P. acnes and inhibitory activity on lipase. ES had no significant cytotoxicity on THP-1 cells. ES suppressed the mRNA levels and production of IL-8, TNF-a, IL-1β in vitro. ES inhibited the expression levels of pro-inflammatory cytokines and the MAPK signaling pathway. Ear thickness and inflammatory cells were markedly reduced by ES treatment. Protocatechuic acid, gallic acid, quercetin, and kaempferol were detected by LC-MS analysis in ES. Conclusions Our results demonstrate antibacterial and anti-inflammatory activities of ES extract against P. acnes. It is suggested that ES extract might be used to treatment anti-inflammatory skin disease. Electronic supplementary material The online version of this article (10.1186/s12906-018-2320-8) contains supplementary material, which is available to authorized users.

The aim of this study was to develop a method for the rapid detection of Gardnerella vaginalis, which is proposed to play a key role in the pathogenesis of bacterial vaginosis. Specific loop-mediated isothermal amplification (LAMP) primers were designed and used to detect target DNA within 45 min under isothermal conditions. Comparative screening indicated that the LAMP assay is superior to PCR in terms of rapidity, and is equivalent in sensitivity and specificity. This LAMP assay can be used for rapid screening and detection of G. vaginalis in vaginal samples; the limit of detection is 10 fg DNA.

The International Space Station (ISS) is a huge manned construct located approximately 400 km above the earth and is inhabited by astronauts performing space experiments. Because the station is within a closed microgravity environment, the astronauts are subject to consistent stress. This study analyzed the temporal changes in the skin fungal microbiota of 10 astronauts using pyrosequencing and quantitative PCR assay before, during, and after their stay in the ISS. Lipophilic skin fungi, Malassezia predominated most samples regardless of the collection period, body site (cheek or chest), or subject. During their stay in the ISS, the level of Malassezia colonization changed by 7.6- ± 7.5-fold (mean ± standard deviation) and 9.5- ± 24.2-fold in cheek and chest samples, respectively. At the species level, M. restricta, M. globosa, and M. sympodialis were more abundant. In the chest samples, the ratio of M. restricta to all Malassezia species increased, whereas it did not change considerably in cheek samples. Fungal diversity was reduced, and the ratio of Malassezia to all fungal colonization increased during the astronauts' stay at the ISS. The ascomycetous yeast Cyberlindnera jadinii was detected in abundance in the in-flight sample of 5 of the 10 astronauts. The microorganism may have incidentally adhered to the skin during the preflight period and persisted on the skin thereafter. This observation suggests the ability of a specific or uncommon microorganism to proliferate in a closed environment. Our study is the first to reveal temporal changes in the skin fungal microbiota of ISS astronauts. These findings will provide information useful for maintaining the health of astronauts staying in the space environment for long periods and for preventing infection due to the human skin microbiota.

Cryptococcus gattii and C. neoformans are pathogenic yeasts that cause meningoencephalitis. C. gattii has four molecular types: VGI, VGII, VGIII, and VGIV. Furthermore, three genotypes have been reported for VGII, and a high pathogenicity of the VGIIa genotype has been proposed. The VGIIa strain has been isolated from a patient in Japan, but little is known about the characteristics of the polysaccharides in this strain. In this study we examined the induction of interleukin-8（IL-8）transcriptional activation and compared the nuclear magnetic resonance（NMR）spectra of extracellular polymeric substances（EPSs）, mainly polysaccharides, from the VGIIa, VGIIb, and VGIIc genotypes. The induction of IL-8 by C. gattii EPSs was weaker than that by C. neoformans EPSs. The anomeric proton signals in the NMR spectra of EPSs obtained from VGII isolates were similar, and the polysaccharides were mainly mannose, xylose, galactose, and glucuronic acid. These results suggest that the extracellular polysaccharides from the VGIIa strain isolated in Japan are almost the same as those from other VGII strains.

A wide variety of microorganisms colonize the human skin and are important to maintaining human health. However, this community is highly sensitive to perturbations, and diseases can develop when the skin microbiome is disrupted by a change in host or environmental conditions. The International Space Station (ISS) is a closed environment, and astronauts on the ISS do not wash their bodies as frequently as when they are on the ground. The maintenance of a balanced skin microbiome is important to overall health, disease prevention, and a high quality of life while on the ISS. The skin fungal microbiome is dominated by Malassezia sp. These lipophilic fungi are ubiquitous across different skin types, whereas changes in the levels of M. globosa and M. restricta are correlated with the formation of seborreich dermatitis/dandruff. The Malassezia microbiome on the skin of astronauts staying on the ISS changed, and there was a reduction in skin fungal microbial diversity. These findings provide useful information about temporal changes in the hygiene of astronauts who are on the ISS for an extended period and indicate that Malassezia microbiome as microbiological markers of skin hygiene.

著者人数: 12名, Accepted: 2015-05-12, 資料番号: SA1150187000

The common skin disease acne vulgaris is caused by Propionibacterium acnes. A lipase secreted by this microorganism metabolizes sebum and the resulting metabolites evoke inflammation in human skin. The antifungal drug ketoconazole inhibits P. acnes lipase activity. We previously showed that the drug also inhibits the growth of P. acnes. Thus, ketoconazole may serve as an alternative treatment for acne vulgaris, which is important because the number of antibiotic-resistant P. acnes strains has been increasing.

Figure S1. The antibacterial effect of ES on skin microbe. To evaluate the antibacterial activity of ES extract against skin microbe, the strains were co-cultured with various concentrations of ES for 48Â h. (A) Staphylococcus aureus, (B) Staphylococcus epidermis, (C) Propionibacterium granulosum. (DOCX 393 kb)

Fungal taxonomy has been reconstructed on the basis of genome information, and new nomenclatural rules have been enacted from 2013. It has been proposed that Cryptococcus neoformans and Cryptococcus gattii be reclassified into two species (C. neoformans and Cryptococcus deneoformans) and five species (C. gattii, Cryptococcus bacillisporus, Cryptococcus deuterogattii, Cryptococcus tetragattii, and Cryptococcus decagattii), respectively. The genus Trichosporon has been reclassified into five genera. Trichosporon asahii, which is the causative agent of trichosporonosis, has been retained in the genus Trichosporon, while Trichosporon cutaneum has been transferred into a new genus, Cutaneotrichosporon.

The common skin disease acne vulgaris is caused by Propionibacterium acnes. A lipase secreted by this microorganism metabolizes sebum and the resulting metabolites evoke inflammation in human skin. The antifungal drug ketoconazole inhibits P. acnes lipase activity. We previously showed that the drug also inhibits the growth of P. acnes. Thus, ketoconazole may serve as an alternative treatment for acne vulgaris, which is important because the number of antibiotic-resistant P. acnes strains has been increasing.

Here, we report the draft genome sequence of Trichosporon domesticum JCM 9580, isolated from the house of a patient with summer-type hypersensitivity pneumonitis (SHP) in Japan. This genomic information will help elucidate the mechanisms of the development of SHP.

The skin surface is colonized by a wide variety of fungi and bacteria. While many of these organisms, including Malassezia, Candida, Streptococcus and Staphylococcus species, are associated with provocation and/or exacerbation of psoriasis, a detailed analysis of the cutaneous fungal microbiome in psoriatic patients has yet to be performed. To identify the disease-specific fungal microbiota on psoriatic scale samples, fungal rRNA gene sequences from 12 psoriatic patients and 12 healthy controls were analyzed by pyrosequencing. A total of 317 806 high-quality sequences were obtained, representing 142 genera. Malassezia species were the most abundant sequences in both populations (46.9 ± 14.0% in psoriasis vs. 76.0 ± 14.6% for healthy controls). Principal coordinate analysis revealed that the fungal microbiomes were independent. Although an association between the cutaneous fungal microbiome and psoriasis has yet to be established, our data indicate that the microbiome in patients with psoriasis is independent of that in healthy controls.

Background: Various types of microorganisms colonize the skin surface, and some such as the bacterium Staphylococcus aureus and fungus Malassezia, exacerbate the symptoms of atopic dermatitis. Malassezia-specific IgE antibodies are present in the sera of patients with atopic dermatitis and the level thereof correlates with the severity of the condition. Malassezia has many genotypes. In the present study, we explored the relationships among the genotypes of Malassezia species colonizing patients with atopic dermatitis, the clinical severity of the disease, and the level of specific IgE antibody. Methods: Scale samples were obtained from head or neck lesions of 74 patients with atopic dermatitis and 40 healthy subjects. The intergenic spacer (IGS) 1 region of the rRNA genes of M. globosa (the major flora of patients with atopic dermatitis) were amplified by PCR and directly sequenced. M. globosa-specific IgE antibody levels were determined using the AlaSTAT™ microplate system. Results: Eighteen M. globosa IGS1 genotypes were detected in scale samples from patients with atopic dermatitis and healthy individuals. Of these, the proportion of the (GT)10:(CT)8 genotype increased with the clinical severity of disease and increasing levels of M. globosa-specific IgE antibodies, whereas this genotype was not found on the skin of healthy subjects. Conclusion: A specific genotype of Malassezia selectively colonized the skin of patients with atopic dermatitis, contributing to the clinical severity of disease. Thus, a “bad Malassezia” may be present on the skin of patients with atopic dermatitis.

Background: Seborrheic dermatitis (SD) is an inflammatory disease associated with seborrhea that appears most often on the face, especially the nasolabial folds, scalp, and upper trunk, in sebaceous gland-rich areas of the skin. Although various factors are involved in the development of SD, Malassezia species of skin fungi play an important role in this skin disease. The aim of this study was to obtain basic information to elucidate the mycological involvement in the development of SD by performing a comprehensive analysis of the skin fungal microbiota of lesional and non-lesional sites in SD patients using a pyrosequencing approach. Methods: Scale samples were collected from lesional and non-lesional sites of 24 Japanese patients with seborrheic dermatitis and the skin fungal microbiome in the scale samples was analyzed using a pyrosequencing method. Results: Thirty taxa were detected from a total of 480,186 high-quality sequences. Malassezia species predominated at both lesional and non-lesional sites; however, the fungal microbiota at lesional sites was more diverse than at non-lesional sites. The fungal communities at lesional and non-lesional sites were clearly distinguished on principal coordinate analysis. Conclusion: Our data indicated that the skin fungal microbiota in lesional sites of SD patients was independent of that in non-lesional sites and will be utilized as basic information for understanding the association between skin fungi and development of SD.

The basidiomycetous yeast Trichosporon faecale is considered to be a non-pathogenic fungus; however, the microorganism has been isolated from clinical specimens in several countries. Trichosporon faecale is classified as type I, II, or III depending on the sequence of the intergenic spacer (IGS) region in its rRNA gene. In this study, 28 T. faecale strains obtained from Japanese subjects and environmental samples were found to be type I. In addition, T. faecale was detected by PCR in 32 scale samples from 146 Japanese healthy subjects, and all 32 samples were found to be type I. Our findings suggest a lack of intraspecific diversity among T. faecale samples originating from Japanese subjects and that T. faecale is part of the skin fungal microbiome.

We previously reported the first isolation of Pseudozyma species from the blood of Thai patients. In this study, three additional new Pseudozyma species were isolated from clinical specimens from Thai patients. The Pseudozyma species showed relatively low sensitivity to azole antifungal agents. The names proposed for these isolates are Pseudozyma alboarmeniaca (DMST 17135(T) = JCM 12454(T ) = CBS 9961(T)), Pseudozyma crassa (DMST 17136(T ) = JCM 12455(T) = CBS 9959(T)) and Pseudozyma siamensis (DMST 17137(T ) = JCM 12456(T) CBS 9960(T)), where DMST is Department of Medical Sciences Culture Collection, JCM is Japan Collection of Microorganisms and CBS is Centraalbureau voor Schimmelcultures.