Distribution of Malassezia species in patients with psoriasis and healthy individuals in Tehran, Iran

Background: Psoriasis is a non‐contagious disorder that affects the skin as red scaly patches. Although the role of Malassezia species in the pathogenesis of psoriasis is still not fully understood, it is thought that these lipophilic yeasts might be a trigger factor in the exacerbation of psoriatic lesions. Methods: Using culture in a specific medium followed by the polymerase chain reaction‐restriction fragment length polymorphism method, the presence of Malassezia species in the skin of 110 patients with psoriasis was compared with that in a control group of 123 healthy patients. Results: The recovery rate of Malassezia species from the skin of patients with psoriasis was significantly lower than that in the controls. In both psoriatic and healthy skin, Malassezia globosa was isolated as the predominant species. In psoriatic patients, the rate of colonization of Malassezia furfur and Malassezia restricta was almost twice that in the controls, whereas M. globosa was isolated more frequently from healthy individuals than from patients. Conclusions: Considering the higher lipase activity secretion by M. furfur in comparison with other Malassezia species, the enzymatic release of arachidonic acid and its metabolites by M. furfur may exacerbate the inflammatory and hyperproliferative changes observed in psoriasis.     

Malassezia yeasts activate the NLRP3 inflammasome in antigen‐presenting cells via Syk‐kinase signalling

Although being a normal part of the skin flora, yeasts of the genus Malassezia are associated with several common dermatologic conditions including pityriasis versicolour, seborrhoeic dermatitis (SD), folliculitis, atopic eczema/dermatitis (AE/AD) and dandruff. While Malassezia spp. are aetiological agents of pityriasis versicolour, a causal role of Malassezia spp. in AE/AD and SD remains to be established. Previous reports have shown that fungi such as Candida albicans and Aspergillus fumigatus are able to efficiently activate the NLRP3 inflammasome leading to robust secretion of the pro‐inflammatory cytokine IL‐1β. To date, innate immune responses to Malassezia spp. are not well characterized. Here, we show that different Malassezia species could induce NLRP3 inflammasome activation and subsequent IL‐1β secretion in human antigen‐presenting cells. In contrast, keratinocytes were not able to secrete IL‐1β when exposed to Malassezia spp. Moreover, we demonstrate that IL‐1β secretion in antigen‐presenting cells was dependent on Syk‐kinase signalling. Our results identify Malassezia spp. as potential strong inducers of pro‐inflammatory responses when taken up by antigen‐presenting cells and identify C‐type lectin receptors and the NLRP3 inflammasome as crucial actors in this process.     

Utilization of matrix‐assisted laser desorption and ionization time‐of‐flight mass spectrometry for identification of infantile seborrheic dermatitis‐causing Malassezia and incidence of culture‐based cutaneous Malassezia microbiota of 1‐month‐old infants

Matrix‐assisted laser desorption and ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS) has been utilized for identification of various microorganisms. Malassezia species, including Malassezia restricta, which is associated with seborrheic dermatitis, has been difficult to identify by traditional means. This study was performed to develop a system for identification of Malassezia species with MALDI‐TOF‐MS and to investigate the incidence and variety of cutaneous Malassezia microbiota of 1‐month‐old infants using this technique. A Malassezia species‐specific MALDI‐TOF‐MS database was developed from eight standard strains, and the availability of this system was assessed using 54 clinical strains isolated from the skin of 1‐month‐old infants. Clinical isolates were cultured initially on CHROMagar Malassezia growth medium, and the 28S ribosomal DNA (D1/D2) sequence was analyzed for confirmatory identification. Using this database, we detected and analyzed Malassezia species in 68% and 44% of infants with and without infantile seborrheic dermatitis, respectively. The results of MALDI‐TOF‐MS analysis were consistent with those of rDNA sequencing identification (100% accuracy rate). To our knowledge, this is the first report of a MALDI‐TOF‐MS database for major skin pathogenic Malassezia species. This system is an easy, rapid and reliable method for identification of Malassezia.     

Phospholipase activity after β‐endorphin exposure discriminates Malassezia strains isolated from healthy and seborrhoeic dermatitis skin

Background Phospholipase activity and its induction by β‐endorphin have been associated with pathogenic Malassezia pachydermatis animal isolates. Objective To evaluate Malassezia phosholipase activity in human isolates from seborrhoeic dermatitis (SD) and healthy controls before and after β‐endorphin exposure. Methods Eighty‐four volunteers with or without SD (N = 41) were sampled. Isolated Malassezia strains were incubated in Dixon’s medium with and without 100 nmol/L β‐endorphin. Subsequently, phospholipase activity was assessed in egg‐yolk agar and the results were compared employing Wilcoxon sign test for paired data, chi‐squared test and multinomial logistic regression analysis. Results A total of 64 Malassezia strains were isolated. SD strains tended to have decreased phospholipase activity before (P = 0.057) and increased after exposure to β‐endorphin (P = 0.061) compared to isolates from healthy skin. Phospholipase activity after β‐endorphin exposure related to basal enzyme activity as a measure of per strain phospholipase inducibility by β‐endorphin did not depend on Malassezia species (P = 0.652). However, this latter biochemical trait discriminates strains isolated from SD lesional and healthy skin (P = 0.036). Conclusion β‐endorphin exposure modifies the in vitro phosholipase activity in Malassezia species isolated from SD lesional skin. This is in accordance with emerging evidence that enhanced local lipase activity is involved in the pathogenesis of SD.     

Effects of Malassezia yeasts on serum Th1 and Th2 cytokines in patients with guttate psoriasis

Background Systemic and focal infections caused by microorganisms have been known to induce or exacerbate psoriasis. Although the role of yeast species of the genus Malassezia in the pathogenesis of psoriasis is not fully understood, it is thought that these lipophilic yeasts may represent a triggering factor in the exacerbation of psoriatic lesions. Objectives This study investigated the effects of Malassezia yeasts on serum Th1 and Th2 cytokines in patients with guttate psoriasis (GP) in order to define their role in the pathogenesis of psoriasis. Methods Fifty patients with GP and 29 clinically healthy individuals were included in the study. All samples consisted of scales and scrapings taken from the scalps, trunks, and upper limbs of both psoriasis patients and healthy subjects. Psoriasis patients and healthy subjects were grouped according to their positivity or negativity for Malassezia yeasts as ascertained by direct microscopy and/or culture. An enzyme‐linked immunosorbent assay (ELISA) was used to measure serum levels of Th1 and Th2 cytokines in these groups. Results No significant differences in positivity for Malassezia yeasts were found between psoriatic skin and healthy skin in samples taken from different body sites. Serum interleukin‐13 (IL‐13) levels were significantly lower in the psoriasis group compared with the control group (P = 0.04). Levels of other cytokines did not differ significantly between the psoriasis and control groups. Mean levels of Th2 cytokines (IL‐4, IL‐10, IL‐13), but not of Th1 cytokines (IL‐2 and IFN‐γ), were significantly lower in psoriasis patients positive for Malassezia yeasts compared with those negative for Malassezia yeasts and control subjects (P = 0.04, P < 0.001 and P = 0.01, respectively). Conclusions The isolation of Malassezia yeasts from GP lesions does not necessarily mean that these species are pathogenic, but their downregulating effects on anti‐inflammatory Th2 cytokines may contribute to the occurrence of GP.     

Seborrheic dermatitis: Etiology,risk factors,and treatments:: Facts and controversies

Seborrheic dermatitis (SD) is a common skin condition seen frequently in clinical practice. The use of varying terms such as sebopsoriasis, seborrheic dermatitis, seborrheic eczema, dandruff, and pityriasis capitis reflects the complex nature of this condition. Despite its frequency, much controversy remains regarding the pathogenesis of SD. This controversy extends to its classification in the spectrum of cutaneous diseases, having being classified as a form of dermatitis, a fungal disease, or an inflammatory disease, closely related with psoriasis. Some have postulated that SD is caused by Malassezia yeasts, based on the observation of their presence in affected skin and the therapeutic response to antifungal agents. Others have proposed that Malassezia is incidental to a primary inflammatory dermatosis that resulted in increased cell turnover, scaling, and inflammation in the epidermis, similar to psoriasis. The presence of host susceptibility factors, permitting the transition of M furfur to its pathogenic form, may be associated with immune response and inflammation. Metabolites produced by Malassezia species, including oleic acid, malssezin, and indole-3-carbaldehyde, have been implicated. SD also has been traditionally considered to be a form of dermatitis based on the presence of Malassezia in healthy skin, the absence the pathogenic mycelial form of Malassezia yeasts in SD, and its chronic course. As a result, proposed treatments vary, ranging from topical corticosteroids to topical antifungals and antimicrobial peptides.     

Superficial veterinary mycoses

Dermatophytes are significant pathogens in animal health due to their zoonotic potential, the economic consequences of infection in farm animal and fur production systems, and the distressing lesions they cause in small domestic pets. Malassezia spp are normal commensal and occasional pathogens of the skin of many veterinary species. Malassezia pachydermatis is a very common cause of otitis and pruritic dermatitis in dogs but is of less importance in other veterinary species.     

La pitiriasis versicolor y las levaduras del género Malassezia

Although pityriasis versicolor is the only human disease for which Malassezia yeasts have been fully established as pathogens, it is still not clear which species are implicated. Most studies carried out in recent years support our hypothesis —proposed in 1999— that Malassezia globosa is the predominant species in pityriasis versicolor lesions, at least in temperate climates. Confirmation of this hypothesis could help us understand the conditions, as yet unclear, that induce transformation of this yeast from the saprophytic form present in healthy skin to the parasitic form, characterized by the formation of pseudomycelium, and could also guide therapy. In addition, isolation of another species, Malassezia furfur, which seems to be predominant in the tropics, raises the possibility of a second etiologic agent confined to certain areas, as occurs with some other human mycoses.     

Skin diseases associated with Malassezia species

The yeasts of the genus Malassezia have been associated with a number of diseases affecting the human skin, such as pityriasis versicolor, Malassezia (Pityrosporum) folliculitis, seborrheic dermatitis and dandruff, atopic dermatitis, psoriasis, and--less commonly--with other dermatologic disorders such as confluent and reticulated papillomatosis, onychomycosis, and transient acantholytic dermatosis. Although Malassezia yeasts are a part of the normal microflora, under certain conditions they can cause superficial skin infection. The study of the clinical role of Malassezia species has been surrounded by controversy because of their fastidious nature in vitro, and relative difficulty in isolation, cultivation, and identification. Many studies have been published in the past few years after the taxonomic revision carried out in 1996 in which 7 species were recognized. Two new species have been recently described, one of which has been isolated from patients with atopic dermatitis. This review focuses on the clinical, mycologic, and immunologic aspects of the various skin diseases associated with Malassezia. It also highlights the importance of individual Malassezia species in the different dermatologic disorders related to these yeasts.     

Comparison of Nested PCR and RFLP for Identification and Classification of Malassezia Yeasts from Healthy Human Skin

Background

Malassezia yeasts are normal flora of the skin found in 75~98% of healthy subjects. The accurate identification of the Malassezia species is important for determining the pathogenesis of the Malassezia yeasts with regard to various skin diseases such as Malassezia folliculitis, seborrheic dermatitis, and atopic dermatitis.

Objective

This research was conducted to determine a more accurate and rapid molecular test for the identification and classification of Malassezia yeasts.

Methods

We compared the accuracy and efficacy of restriction fragment length polymorphism (RFLP) and the nested polymerase chain reaction (PCR) for the identification of Malassezia yeasts.

Results

Although both methods demonstrated rapid and reliable results with regard to identification, the nested PCR method was faster. However, 7 different Malassezia species (1.2%) were identified by the nested PCR compared to the RFLP method.

Conclusion

Our results show that RFLP method was relatively more accurate and reliable for the detection of various Malassezia species compared to the nested PCR. But, in the aspect of simplicity and time saving, the latter method has its own advantages. In addition, the 26S rDNA, which was targeted in this study, contains highly conserved base sequences and enough sequence variation for inter-species identification of Malassezia yeasts.     

Molecular Analysis of Malassezia Microflora on the Skin of the Patients with Atopic Dermatitis

Background

The yeasts of the genus Malassezia are members of the normal flora on human skin and they are found in 75~80% of healthy adults. Since its association with various skin disorders have been known, there have been a growing number of reports that have implicated Malassezia yeast in atopic dermatitis (AD).

Objective

The aim of the present study is to isolate the various Malassezia species from AD patients by using 26S rDNA (ribosomal Deoxyribonucleic acid) PCR-RFLP and to investigate the relationship between a positive Malassezia culture and the severity of AD.

Methods

Cultures for Malassezia yeasts were taken from the scalp, cheek, chest, arm and thigh of 60 patients with atopic dermatitis. We used a rapid and accurate molecular biological method 26S rDNA PCR-RFLP, and this method can overcome the limits of the morphological and biochemical methods.

Results

Positive Malassezia growth was noted on 51.7% of the patients with atopic dermatitis by 26S rDNA PCR-RFLP analysis. The overall dominant species was M. sympodialis (16.3%). M. restricta was the most common species on the scalp (30.0%) and cheek (16.7%). M. sympodialis (28.3%) was the most common species on the chest. The positive culture rate was the highest for the 11~20 age group (59.0%) and the scalp showed the highest rate at 66.7%. There was no significant relationship between the Malassezia species and SCORing for Atopic Dermatitis (SCORAD).

Conclusion

The fact that the cultured species was different for the atopic dermatitis lesion skin from that of the normal skin may be due to the disrupted skin barrier function and sensitization of the organism induced by scratching in the AD lesion-skin. But there was no relationship between the Malassezia type and the severity score. The severity score is thought to depend not on the type, but also on the quantity of the yeast.     

The role of the skin microbiome in atopic dermatitis: a systematic review

Dysbiosis is a hallmark of atopic dermatitis (AD). The composition of skin microbiome communities and the causality of dysbiosis in eczema have not been well established. The objective of this review is to describe the skin microbiome profile in AD and address whether there is a causal relationship between dysbiosis and AD. The protocol is registered in PROSPERO (CRD42016035813). We searched PubMed, Embase, Scopus and ClinicalTrials.gov for primary research studies applying culture‐independent analysis on the microbiome on AD skin of humans and animal models. Two authors independently screened the full text of studies for eligibility and assessed risk of bias. Because of heterogeneity no quantitative synthesis was done. Of 5735 texts, 32 met the inclusion criteria (17 published: 11 human and six animal studies). The studies varied in quality and applied different methodology. The skin in AD had low bacterial diversity (lowest at dermatitis‐involved sites) and three studies showed depletion of Malassezia spp. and high non‐Malassezia fungal diversity. The relative abundance of Staphylococcus aureus and Staphylococcus epidermidis were elevated and other genera were reduced, including Propionibacterium. A mouse study indicated that dysbiosis is a driving factor in eczema pathogenesis. The data are not sufficiently robust for good characterization; however, dysbiosis in AD not only implicates Staphylococcus spp., but also microbes such as Propionibacterium and Malassezia. A causal role of dysbiosis in eczema in mice should encourage future studies to investigate if this also applies to humans. Other important aspects are temporal dynamics and the influence of methodology on microbiome data.     

Malassezia globosa tends to grow actively in summer conditions more than other cutaneous Malassezia species

Malassezia globosa is a major pathogen of Malassezia folliculitis (MF) and the predominant species on human skin. The aim of this study was to clarify the differences between M. globosa and other cutaneous Malassezia species, M. restricta, M. dermatis, M. sympodialis and M. furfur. The optimum growth temperature, effects of compounds of sweat and free fatty acids on growth, and lipase activities of five cutaneous Malassezia species were determined. The growth of M. globosa was promoted strongly by the compounds of sweat and high temperature unlike that of other cutaneous Malassezia species. This result clarified that M. globosa tended to grow actively in summer conditions more than other cutaneous Malassezia species. Furthermore, M. globosa showed high lipase activity. We consider these characteristics of M. globosa to relate to the pathogenesis of MF.     

Progress in Malassezia Research in Korea

Yeasts of the genus Malassezia are part of the normal flora of human skin. However, they are also associated with various skin diseases. Since the introduction of Malassezia to the Korean Dermatologic Society two decades ago, remarkable progress has been made in our knowledge of this genus. In this paper, we review recent developments in Malassezia research, including taxonomy and methods for species identification, recent genome analyses, Malassezia species distribution in healthy conditions and in specific skin diseases, trials investigating the mechanisms underlying Malassezia-related diseases, as well as therapeutic options. This review will enhance our understanding of Malassezia yeasts and related skin diseases in Korea.     

The Investigation on the Distribution of Malassezia Yeasts on the Normal Korean Skin by 26S rDNA PCR-RFLP

Background

Malassezia yeasts are normal flora of the skin that are discovered in 75~98% of health subjects, but since its association with various skin disorders have been known, many studies have been conducted in the distribution of the yeasts.

Objective

To isolate, identify, and classify Malassezia yeasts from the normal human skin of Koreans by using the rapid and accurate molecular biology method (26S rDNA PCR-RFLP) which overcome the limits of morphological and biochemical methods, and to gather a basic database that will show its relation to various skin diseases.

Methods

Malassezia yeasts were cultured from clinically healthy human skin using scrub-wash technique at five sites (forehead, cheek, chest, upper arm, and thigh) and swabbing technique at scalp in 160 participants comprised of 80 males and 80 females aged from 0 to 80. Identification of obtained strains were placed into the one of the 11 species by 26S rDNA PCR-RFLP.

Results

An overall positive culture rate was 62.4% (599/960). As shown in the experiment groups by their age, the positive culture rate was the highest (74.2%) in the age 21~30 and 31~40 (89/120). In the experiment groups by different body areas, the scalp showed the highest positive culture rate of 90% (144/160). On analysis of 26S rDNA PCR-RFLP, M. globosa was the most predominant species in the age 0~10 (32.8%), 11~20 (28.9%), 21~30 (32.3%). M. restricta was identified as predominant species in the age 41~50 (27.9%), 61~70 (31.5%) and 71~80 (24.0%). In the age 31~40 years, M. sympodialis was found to be the most common species (24.6%). According to body site, M. restricta was more frequently recovered in the scalp (56.8%), forehead (39.8%) and cheek (24.0%) and while M. globosa was more frequently recovered in the chest (36.8%). Higher positive culture rates of Malassezia yeasts were shown in male subjects than female counterparts in all body areas except scalp (p<0.05). Especially in this study, M. dermatis, newly isolated Malassezia species from atopic dermatitis patient in Japan, was isolated and identified in 19 cases (1.9%) in healthy subjects.

Conclusion

The key is to recognize the existence of a difference in the type of Malassezia species in different ages as well as body areas, which reflects differing skin lipid levels in various ages and different body areas. Moreover, 26S rDNA PCR-RFLP analysis which was opted in this study could provide a sensitive and rapid identification system for Malassezia species, which may be applied to epidemiological surveys and clinical practice.     

The prevalence of Malassezia yeasts in patients with atopic dermatitis, seborrhoeic dermatitis and healthy controls

Cultures for Malassezia yeasts were taken from both normal-looking skin and lesional skin in 124 patients with atopic dermatitis, 16 patients with seborrhoeic dermatitis and from normal skin of 31 healthy controls. Positive Malassezia growth was found in fewer patients with atopic dermatitis (56%) than in patients with seborrhoeic dermatitis (88%) or in healthy controls (84%, p<0.01). In the patients with atopic dermatitis, fewer positive cultures were found in lesional (28%) than in non-lesional skin (44%, p<0.05), while positive cultures were found in 75% of both lesional and non-lesional skin of patients with seborrhoeic dermatitis (not significant). M. sympodialis dominated in patients with atopic dermatitis (46%) and in healthy controls (69%). In patients with seborrhoeic dermatitis both M. sympodialis and M. obtusa were cultured in 43%. A Malassezia species extract mixture would increase the possibility of detecting IgE sensitization to Malassezia in patients with atopic dermatitis.     

Malassezia yeasts and their significance in dermatology

Yeasts of the genus Malassezia belong to the normal microflora of the human skin. In addition they are known to cause a variety of skin diseases; the most frequent of which is pityriasis versicolor. Malassezia yeasts are also thought to be associated with seborrheic dermatitis, dandruff and Malassezia folliculitis. Recently the significance of Malassezia yeasts as a trigger factor for atopic dermatitis of the head and neck region has been pointed out. The role of the Malassezia yeasts in these different diseases has been controversial in the past and remains an issue because of difficulties in isolation, culture and differentiation of the organism. Thanks to molecular techniques, 10 species can actually be differentiated. The article presents the different Malassezia-associated diseases, their clinical picture, diagnosis and appropriate therapy. In addition the speciation of Malassezia is reviewed.     

Malassezia-Hefen und ihre Bedeutung in der Dermatologie

Yeasts of the genus Malassezia belong to the normal microflora of the human skin. In addition they are known to cause a variety of skin diseases; the most frequent of which is pityriasis versicolor. Malassezia yeasts are also thought to be associated with seborrheic dermatitis, dandruff and Malassezia folliculitis. Recently the significance of Malassezia yeasts as a trigger factor for atopic dermatitis of the head and neck region has been pointed out. The role of the Malassezia yeasts in these different diseases has been controversial in the past and remains an issue because of difficulties in isolation, culture and differentiation of the organism. Thanks to molecular techniques, 10 species can actually be differentiated. The article presents the different Malassezia-associated diseases, their clinical picture, diagnosis and appropriate therapy. In addition the speciation of Malassezia is reviewed.     

Analysis of the response of human keratinocytes to Malassezia globosa and restricta strains

Malassezia spp. are saprophyte yeasts involved in skin diseases with different degrees of severity. The aim of our study was to analyze the response of human epidermal keratinocytes to Malassezia globosa and restricta strains evaluating the host defence mechanisms induced by Malassezia spp. colonization. Our results showed a different modulation of the inflammatory and immunomodulatory cytokine pathways obtained with the different strains of Malassezia tested. In addition, this expression is altered by blocking the TLR2 receptor. In comparison with M. furfur, M. globosa and restricta displayed an unexpected and striking cytotoxicity on keratinocytes. The differences observed could be related to the different modalities of interaction between keratinocytes and Malassezia strains, but also to their growth condition. Taken together, these results indicate that M. globosa or M. restricta colonization exert a different control on the cytokine inflammatory response activated in the human keratinocyte in which TLR2 might be involved. M. globosa and M. restricta may play a synergistic role in the exacerbation of skin diseases in which both are found.