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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.

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.

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.

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.

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.

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.

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.

SummaryThe fungus Trichosporon asahii is distributed widely in the environment and is frequently isolated from clinical specimens. It can cause life-threatening disseminated infections, including breakthrough infection after the administration of candin antifungal drugs. However, the definitive infection route is still unclear. In this study, 43 T. asahii genotypes were detected in 72 fecal samples from healthy subjects. The rRNA genotypes of the species were determined using a culture-independent method. The genotypes of T. asahii were almost identical to those of reported clinical isolates. Therefore, T. asahii colonizing the gastrointestinal tract might be associated with the development of trichosporonosis, and not environmental isolates.

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.