Taxonomy of Malassezia furfur: state of the art

Malassezia furfur is a lipophilic yeast considered as a normal component of the human skin flora. Apart from pityriasis versicolor, M. furfur has been linked to several skin diseases such as seborrheic dermatitis, folliculitis or atopic dermatitis. Moreover, these yeasts have been reported as agent of invasive human diseases including pneumonia, catheter-associated sepsis and peritonitis. The existence of morphological, serological, metabolical, biochemical and karyotipical differences has been described among isolates of these yeasts. These observations gave arguments for a possible intraspecific division, and this hypothesis has been confirmed by the existence of six species within the formerly called M. furfur (lipid-dependent Malassezia strains): M. furfur, Malassezia sympodialis, Malassezia globosa, Malassezia obtusa, Malassezia restricta and Malassezia slooffiae.     

Concordance between phenotypical features and PCR-REA for the identification of Malassezia spp

The genus Malassezia has been recently revised and nowadays includes 11 species that cannot always be differentiated from each other by physiological and morphological tests. This study was aimed to evaluate the correlation between a molecular method and conventional phenotypic features in the identification of Malassezia spp. To achieve this aim, 92 Argentinean clinical strains isolated between 2001 and 2005 were analyzed along with three reference strains (Malassezia furfur CBS 7019, Malassezia sympodialis CBS 7222 and Malassezia slooffiae CBS 7956). By using PCR and restriction enzyme analysis with three different DNA endonucleases (PCR-REA), the molecular method consistently identified all three reference strains and all 92 clinical isolates as follows: 63 M. sympodialis, 18 M. furfur, 10 Malassezia globosa and one Malassezia obtusa. Phenotypic studies undentified 85 clinical isolates and two of the reference strains (total agreement > 91%). In particular for M. sympodialis, M. furfur and M. globosa, the species more frequently involved in human pathology, the agreement ranged between 84 and 96%. This result suggests that phenotypic studies are suitable for the presumptive identification of important Malassezia species in the clinical medical mycology laboratories where molecular methodologies are not available.     

Chitin synthase 2 gene sequence of Malassezia species.

Nucleotide sequences of the chitin synthase 2 (CHS2) gene of seven species, Malassezia furfur, M. globosa, M. obtusa, M. pachydermatis, M. restricta, M. slooffiae and M. sympodialis, were analyzed for their phylogenetic relationship. About 620-bp genomic DNA fragments of the CHS2 gene were amplified from these Malassezia species by polymerase chain reaction (PCR) and sequenced. The CHS2 nucleotide sequences of these Malassezia species showed more than 95% similarity between the species. A phylogenetic analysis of the nucleotide sequences of CHS2 gene fragments of seven Malassezia species revealed that the species were genetically distinct from each other.     

正常人耵聍中马拉色菌菌种构成研究

目的研究正常人耵聍中马拉色菌菌种构成及同一宿主耵聍中菌种是否一致。方法采集45名健康志愿者双侧耵聍,0.1%曲拉通X-100溶解稀释后接种于含菜籽油培养基,生化及形态学方法鉴定到种,同时提取菌种DNA,用真菌通用引物ITS1/ITS4做PCR扩增并测序鉴定。结果有44例(97.78%)双侧耵聍中均培养出马拉色菌(共分离出88株菌),菌种构成:糠粃马拉色菌29株(32.95%)、斯洛菲马拉色菌23株(26.14%)、合轴马拉色菌18株(20.45%)、球形马拉色菌11株(12.50%)、限制性马拉色菌7株(7.95%)。44例(88株菌)中双侧耵聍菌种相同者有38例(76株菌)(一致率86.36%)。结论正常人耵聍中马拉色菌菌种分布较广,主要菌种为糠秕马拉色菌。同一宿主双侧耵聍中马拉色菌菌种具有一定的一致性。     

Characterization of Malassezia microbiota in the human external auditory canal and on the sole of the foot

Of the fungal skin microbiota, the lipophilic yeast genus Malassezia predominates at all body sites. Of the members of this genus, M. globosa, M. restricta, and M. sympodialis are the most common on the face, limbs, and trunk. In the present study, the Malassezia microbiotas in the external auditory canal and on the sole of the foot were characterized. M. slooffiae was the most common species in both the external auditory canal and on the sole of the foot, followed by M. restricta. Principal component analysis further revealed that the Malassezia microbiota in the external auditory canal and on the sole of the foot constitute a different cluster from those on the scalp and cheek and in the nasal cavity. Additionally, five new Malassezia phylotypes were detected on the sole of the foot and in the external auditory canal. Our results suggest that a distinctive Malassezia microbiota is present in the external auditory canal and on the sole of the foot, although the clinical significance of this finding remains unknown.     

Detection and quantification of specific IgE antibodies against eight Malassezia species in sera of patients with atopic dermatitis by using an enzyme-linked immunosorbent assay

The lipophilic yeast Malassezia, a member of the cutaneous microflora, is an exacerbating factor in atopic dermatitis (AD). Of the 11 currently recognized species, M. globosa and M. restricta are found to frequently colonize the skin of AD patients. In this study, we attempted to quantify specific IgE antibodies against eight Malassezia species, namely, M. dermatitis, M. furfur, M. globosa, M. obtusa, M. pachydermatis, M. slooffiae, M. sympodialis, and M. restricta, in sera from AD patients by using an enzyme-linked immunosorbent assay (ELISA). The specific IgE value against M. restricta was greater than those against other Malassezia species. Competitive ELISA inhibition tests revealed that M. restricta contained species specific as well as shared antigens. Therefore, M. restricta could be considered as a candidate diagnostic antigen for detecting anti-Malassezia IgE in sera from AD patients.     

The genus Malassezia: old facts and new concepts

Lipophilic yeasts are being considered as major opportunistic pathogens for a very long time. Most of the yeasts show an absolute requirement for long fatty acid chains and specific procedures are required for their isolation, conservation and identification. For that reason, the history of the nomenclature used for the Malassezia genus is quite complex. Before 1996, only 3 species were recognized: Malassezia furfur, M. pachydermatis and M. sympodialis. To date, the genus is composed of one non lipid-dependent species (M. pachydermatis) and 12 lipid-dependent species. No doubt that additional new taxa will be described in close future. Very recently the genome and secretory proteome of two Malassezia species was described. This analysis demonstrated the presence of multiple secreted lipases to aid in harvesting host lipids. It also revealed the presence of mating-type genes, providing an indication that Malassezia yeasts may be capable of sex.     

In vitro modulation of human keratinocyte pro- and anti-inflammatory cytokine production by the capsule of Malassezia species

Malassezia spp. are commensal, cutaneous fungi that are implicated in seborrhoeic dermatitis. We hypothesize that the lipid-rich capsule of Malassezia spp. masks the organism from host detection, and depletion of this layer elicits an inflammatory response. To test this, preparations of capsulated or acapsular [10% (v/v) Triton X-100 treated], viable and nonviable, exponential or stationary phase Malassezia furfur, Malassezia globosa, Malassezia obtusa, Malassezia restricta, Malassezia slooffiae and Malassezia sympodialis, were incubated with normal human keratinocytes. Proinflammatory (IL-6, IL-8, IL-1alpha and tumour necrosis factor-alpha) and anti-inflammatory cytokine (IL-10) release and intracellular IL-10 concentrations were quantified using enzyme-linked immunosorbent assays. Capsulated Malassezia yeasts stimulated limited or no production of inflammatory cytokines, and increased intracellular IL-10 (P < 0.05). Removal of the capsule of many Malassezia preparations caused a significantly increased production of IL-6, IL-8 and IL-1alpha, and a decrease in intracellular IL-10. Notably, acapsular viable, stationary phase M. globosa caused a 66-fold increase in IL-8 production (P < 0.001) and acapsular nonviable, stationary phase M. furfur caused a 38-fold increase in IL-6 production (P < 0.001) and a 12-fold decrease in intracellular IL-10 (P < 0.001). These results support the hypothesis that the lipid layer of Malassezia spp. modulates cytokine production by keratinocytes. This has implications in the pathogenesis of seborrhoeic dermatitis.     

Prevalence of Malassezia species in pityriasis versicolor lesions in northeast Argentina

BackgroundMalassezia species normally colonize the skin but they can change their saprophytic state and invade the stratum corneum as pathogens.AimsTo determine the prevalence of Malassezia species isolated from patients with pityriasis versicolor (PV) and to analyse their distribution according to the location of the lesion on the body.MethodsThis study included 218 patients with PV and positive Malassezia cultures who resided in the city of Resistencia, a subtropical area located in northeast Argentina. Age, gender, and the body site of lesions were recorded. Strains were identified by PCR-RFLP.ResultsMalassezia sympodialis (37.7%) and Malassezia globosa (37.2%) were the most prevalent species isolated alone or in association with other Malassezia species in 82% of the patients. Malassezia furfur (21.3%) was the third most common species, followed by Malassezia slooffiae (1.7%), and Malassezia restricta (1.3%), which was found only in combination with M. globosa and M. sympodialis. Malassezia dermatis (0.4%) and Malassezia pachydermatis (0.4%) were each isolated once. None of the species affected a body site with statistical significance. Significant difference between genders according to age was found only in the 31–40-year-age group.ConclusionsThis study suggests that M. sympodialis and M. globosa represent the main species implicated in the pathogenicity of PV. M. furfur appears to be the third agent of importance in this geographical area. Statistical analyses showed none of the species was particularly associated with any one of the body sites.     

东南亚青年人群中马拉色菌菌种构成分析

目的：研究部分东南亚国家青年人群中马拉色菌菌种构成。方法采集285名青年人（分别来源于中国、尼泊尔、印度、巴基斯坦）面部正常皮肤及皮损区标本,接种于 Leeming ＆amp; Notman 培养基后进行培养分离,生化及形态学方法鉴定到种。结果共分离出8个菌种,共计501株马拉色菌,以球形马拉色菌为主,占40.5℅（203/501）,其次为合轴马拉色菌19.0℅（95/501）、糠秕马拉色菌16.0℅（80/501）、限制马拉色菌11.0℅（55/501）、斯洛菲马拉色菌6.2℅（31/501）等。各国家间未见明显的地理学差异。结论东南亚地区青年正常人群及马拉色菌相关疾病患者中的主要菌种为球形马拉色菌。     

Pathological and clinical aspects of the diseases caused by Malassezia species

From veterinary point of view Malassezia pachydermatis has the greatest significance. It has been standing in the focus of interest since the early 1990s, mostly because of the frequency of otitis externa and dermatitis caused by this yeast in dogs. This is the only lipid-independent species in the genus Malassezia. It can be found in very large proportion on the skin of healthy animals, but can be isolated in much greater number from diseased dogs. It often causes illness together with other pathogens (e.g. Staphylococcus intermedius). Some breeds are predisposed. In addition to the treatment of the accidental concurrent diseases, therapy consists of systemic and/or topical antimicrobial treatment. Ketoconazole is used most frequently. Malassezia pachydermatis plays also a role in the skin disorders of other carnivores. It has little zoonotic potential, it can be dangerous to immunocompromised humans. The other Malassezia species have little veterinary importance, although M. sympodialis and M. globosa were isolated from asymptomatic animals (mostly cats) and from mixed infections.     

The influence of newly synthesised fenporpimorph derivatives on some pathogen yeasts

The effect of minimum inhibitory concentrations (MICs) of six novel fenpropimorph derivatives on lipid and sterol composition of Candida albicans, Cryptococcus neoformans, Malassezia pachydermatis and Malassezia furfur was investigated. The MICs for the most effective derivatives were found in the range from 3.7 to 56.7 microM and were 2-3 times lower compared to the commercial fungicide bifonazol. The more efficient fenpropimorph derivatives were the piperidine derivative for C. albicans and the allylamine derivative for Cr. neoformans, M. pachydermatis and M. furfur. The inhibitor in the growth medium reduced the unsaturation index of the total lipid content in M. furfur and C. albicans.     

江西地区花斑癣患者马拉色菌菌种分析

目的了解江西地区花斑癣患者马拉色菌菌种分布情况。比较传统吐温试验和改良吐温试验。方法对141例临床典型、真菌镜检阳性的花斑癣患者,采用Leeming和Notman培养基培养皮屑。以标准菌株为对照,按形态学和生理生化特点进行分类,分析马拉色菌菌种构成情况。同时比较传统吐温试验和改良吐温试验优缺点。结果培养到95株马拉色菌,分离出5个菌种:合轴马拉色菌62株,糠秕马拉色菌17株,球形马拉色菌9株,钝形马拉色菌6株,限制马拉色菌1株。结论合轴马拉色菌占有明显优势(65.26%)。改良吐温试验易于操作、费时短,尤其适合多标本的流行病学调查研究,值得推广。     

A new yeast, Malassezia yamatoensis, isolated from a patient with seborrheic dermatitis, and its distribution in patients and healthy subjects

Over the last few years, new Malassezia species have been found regularly in Japanese subjects. We isolated another new Malassezia species from a Japanese patient with seborrheic dermatitis (SD), and named it M. yamatoensis. In its physiological characteristics and the utilization of Tween by M. yamatoensis is similar to that of M. furfur and M. dermatis. It is distinguished by its growth temperature. To examine the distribution of the microorganism in the skin of patients with SD and atopic dermatitis (AD), and healthy subjects, we applied transparent dressings to the skin, and detected M. yamatoensis DNA using a non-culture-based method that consisted of nested PCR with specific primers. M. yamatoensis DNA was detected from 3 of 31 SD patients (9.7%), 5 of 36 AD patients (13.9%), and 1 of 22 healthy subjects (4.6%). Therefore, M. yamatoensis is a rare member of the cutaneous microflora.     

Identification of atypical strains of Malassezia spp. from cattle and dog

Yeast species in the genus Malassezia are lipophilic with the exception of Malassezia pachydermatis. During a study of the occurrence of Malassezia species in the external ear of 964 cattle and 6 dogs in Minas Gerais, Brazil, six lipid-dependent isolates could not be identified to known species. Four isolates came from healthy cows, one from a cow with otitis, and one from a healthy dog. When tested with Tweens and Cremophor EL as single sources of lipids, the strains grew on all sources except Cremophor EL. None of the six strains hydrolyzed esculin, and all produced catalase. Pigment production from tryptophan was variable. Partial large subunit rRNA sequences were obtained for two isolates that remained viable in culture. The strain from the cow with otitis was identified as a lipid-dependent variant of M. pachydermatis, and the strain from the dog was an atypical variant of Malassezia furfur.     

南海海军部队特发性瘙痒症患者皮肤马拉色菌定植调查

目的研究南海海军部队特发性皮肤瘙痒症患者皮肤马拉色菌的检出以及菌种的构成,探讨马拉色菌与特发性瘙痒症之间的相关性。方法对采自56例特发性瘙痒症患者与50例健康者的鳞屑标本进行真菌学镜检及采用菜子油培养基培养,根据菌落形态和生理生化特点进行菌种鉴定。结果观察组与对照组镜检阳性率分别为74．10％和72．00％,X2=0．344,P〉0．05两者比较无统计学意义。而镜检马拉色菌孢子密度Ⅲ级以上（每10个高倍视野≥50个者）,观察组与对照组分别为74．70％和38．89％,X2=20．30,P〈0．01。两者统计学比较差异有显著性意义。观察组168份标本共分离出143株马拉色菌,阳性率85．12％。其中合轴马拉色菌97株（67．83％）,球形马拉色菌26株（18．18％）,糠秕马拉色菌19株（13．29％）,钝形马拉色菌1株（0．70％）。对照组分离出马拉色菌107株,阳性率71．33％;其中糠秕马拉色菌55株（51．40％）,合轴马拉色菌33株（30．84％）,球形马拉色菌14株（13．08％）,钝形马拉色菌5株（4．67％）。菌种检出率和菌种构成差异均有显著性（X2=2．99,P〈0．01和7。X2=51.16,P〈0．01）。结论南海海军部队特发性瘙痒症患者皮肤马拉色菌载量较正常人高,其优势菌是合轴马拉色菌。     

Two new lipid-dependent Malassezia species from domestic animals

During a study on the occurrence of lipid-dependent Malassezia spp. in domestic animals, some atypical strains, phylogenetically related to Malassezia sympodialis Simmons et Guého, were shown to represent novel species. In this study, we describe two new taxa, Malassezia caprae sp. nov. (type strain MA383=CBS 10434), isolated mainly from goats, and Malassezia equina sp. nov. (type strain MA146=CBS 9969), isolated mainly from horses, including their morphological and physiological characteristics. The validation of these new taxa is further supported by analysis of the D1/D2 regions of the 26S rRNA gene, the ITS1-5.8S-ITS2 rRNA, the RNA polymerase subunit 1 and chitin synthase nucleotide sequences, and the amplified fragment length polymorphism patterns, which were all consistent in separating these new species from the other species of the genus, and those of the M. sympodialis species cluster, specifically.     

Epidemiology and variability of Malassezia spp

A short review on Malassezia spp., completed with our experience, is made. The main epidemiological characteristics with particular regard to the diffusion in several animal species, the characteristics of skin colonization (in particular of the dog) and the distribution of the different Malassezia spp. in some hosts are discussed. Lastly the main phenotypic and genotypic characteristics, referred to M. pachydermatis especially, were described, showing their high variability and differentiation.     

Suitability of lipid materials for culture ofMalassezia as evaluated from its cellular fatty acid composition

Malassezia is a facultative or obligatory lipophilic yeast. We devised new lipid-supplemented media suitable for the culture ofMalassezia. Malassezia furfur andM. pachydermatis grew well on both solid and liquid media supplemented with creaming powder preparations which are commercially available at moderate prices. Striking differences were found between the cellular fatty acid compositions ofM. furfur grown on media supplemented with creaming powder and that grown on media with conventional olive oil.Malassezia furfur grown on media with olive oil had nearly the same fatty acid composition as olive oil, with C18:1 amounting to 80%, while that grown on media supplemented with creaming powder had C16, C18:1 and C18:2 as the principal components. The use of these supplementary lipids appeared not to inhibit the normal synthesis of fatty acid inM. furfur. For the culture ofM. pachydermatis, media supplemented with creaming powder were also found more suitable than lipid-free media. The media devised are considered excellent, because they appear to provide a more natural growth environment forMalassezia.     

Molecular Epidemiology of Malassezia globosa and Malassezia restricta in Sudanese Patients with Pityriasis Versicolor

Pityriasis versicolor is a superficial infection of the stratum corneum caused by Malassezia yeasts. The cutaneous Malassezia globosa and Malassezia restricta in Sudanese patients with pityriasis versicolor were elucidated using a molecular-based, culture-independent method and compared with that in healthy individuals. Scale samples were collected by applying an Opsite? transparent dressing to lesional and non-lesional sites on 29 Sudanese patients with pityriasis versicolor and 54 healthy individuals. Malassezia DNA was extracted directly from the samples. The overall level of colonization by Malassezia globosa and Malassezia restricta was analyzed by real-time PCR using a TaqMan probe. The overall level of colonization by Malassezia at the lesional sites was higher than that at the non-lesional sites for all body sites, including the face, neck, cheeks, and trunk (2.7- to 6.0-fold increase). Both M. globosa and M. restricta were detected in patients and healthy individuals. However, M. globosa predominated at lesional sites, whereas the level of colonization by both species was similar in healthy individuals.