Isolation of Malassezia globosa and M. sympodialis from patients with pityriasis versicolor in Spain

Pityriasis versicolor is a superficial infection of the stratum corneum by several yeast species formerly collectively named Malassezia furfur. The genus Malassezia has been recently enlarged with new species. With the exception of M. pachydermatis, the remaining six species have an absolute requirement in vitro for supplementation of long-chain fatty acids in media. These lipophilic yeasts comprise six species: M. furfur, M. globosa, M. obtusa, M. restricta, M. slooffiae and M. sympodialis. The aim of this study was to establish whether there was any association between the various species of Malassezia and pityriasis versicolor lesions. Thus, we studied the isolates from 79 patients with pityriasis versicolor, both from lesions and from apparently healthy skin close to the visible lesions. In pityriasis versicolor lesions, the species most frequently isolated was M. globosa (90%), followed by M. sympodialis (41%). Almost all isolates (99%) belonged to one of these two species. The most frequent pattern was M. globosa as the sole species (58% of cases), although the association with M. sympodialis was also frequent (30%). These results confirmed M. globosa as the main agent of pityriasis versicolor and M. sympodialis as the second agent in importance. Malassezia globosa was found to be a species with high levels of esterase and lipase enzymes of probable importance in their pathogenicity. This revised version was published online in June 2006 with corrections to the Cover Date.     

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.     

Identification of Malassezia Species in Patients with Seborrheic Dermatitis in China

Background

The causes of seborrheic dermatitis (SD) are complex and incompletely understood. Among the factors, Malassezia yeasts have been reported to play a major etiological role in SD. Many previous studies adopted conventional culture methods that were disadvantaged to detect Malassezia microflora in SD patients, resulting in a low detection rate for each species and high variance in types of microflora observed.

Objective

This study analyzed Malassezia microflora in SD patients by applying a transparent dressing to the lesional skin and using direct detection of fungal DNA using nested PCR.

Methods

We collected samples from the lesional skin of 146 SD patients in China and extracted fungal DNA directly from the lesional samples without culture. Specific primers for each Malassezia species were designed to amplify existing yeasts in each sample. Some samples were randomly selected to culture and identified by morphological and physiologic criteria.

Results

M. globosa and M. restricta were found in 87.0 and 81.5 % of seborrheic dermatitis patients, respectively, which together accounted for more than 50 % of Malassezia spp. recovered in these Chinese patients. The majority of SD patients (82.9 %) showed co-colonization of two or more Malassezia species.

Conclusion

M. globosa and M. restricta predominated in Malassezia colonization in Chinese SD patients. Compared with conventional culture, non-culture-based methods may more accurately reflect Malassezia microflora constitution.     

Prevalence of Malassezia species in patients with pityriasis versicolor in Rosario, Argentina

BackgroundMalassezia species are considered opportunistic yeasts of increasing clinical importance. These lipophilic yeasts are associated with various human diseases, especially pityriasis versicolor (PV), a chronic superficial scaling dermatomycosis.AimsThe aim of this study was to isolate, identify and analyze the distribution of the different species of Malassezia in patients with PV in Rosario city (Argentina).MethodsA total of 264 clinical samples were studied. Isolates were identified on the basis of microscopic observation of cells, and physiological properties, such as the presence of catalase, ability to use Tween compounds, splitting of esculin, and morphology, color and precipitate production on chromogenic agar CHROMagar-Malassezia medium (CHROMM).ResultsThe highest prevalence of PV in this study was observed in the 25- to 45-year-old group. No differences were found in the development of PV between sexes. The most affected areas of body were the trunk and face. Malassezia sympodialis (51%) was the most commonly isolated species, followed in frequency by M. globosa (40%), Malassezia furfur (7%), Malassezia obtusa (1%) and Malassezia slooffiae (1%).ConclusionsThe success for a correct identification of these yeasts is important to improve our knowledge about their epidemiological role in PV and also to detect the appearance of strains which are resistant to the commonly used antifungal drugs.     

Chemical Organization of the Cell Wall Polysaccharide Core of Malassezia restricta

Malassezia species are ubiquitous residents of human skin and are associated with several diseases such as seborrheic dermatitis, tinea versicolor, folliculitis, atopic dermatitis, and scalp conditions such as dandruff. Host-Malassezia interactions and mechanisms to evade local immune responses remain largely unknown. Malassezia restricta is one of the most predominant yeasts of the healthy human skin, its cell wall has been investigated in this paper. Polysaccharides in the M. restricta cell wall are almost exclusively alkali-insoluble, showing that they play an essential role in the organization and rigidity of the M. restricta cell wall. Fractionation of cell wall polymers and carbohydrate analyses showed that the polysaccharide core of the cell wall of M. restricta contained an average of 5% chitin, 20% chitosan, 5% β-(1,3)-glucan, and 70% β-(1,6)-glucan. In contrast to other yeasts, chitin and chitosan are relatively abundant, and β-(1,3)-glucans constitute a minor cell wall component. The most abundant polymer is β-(1,6)-glucans, which are large molecules composed of a linear β-(1,6)-glucan chains with β-(1,3)-glucosyl side chain with an average of 1 branch point every 3.8 glucose unit. Both β-glucans are cross-linked, forming a huge alkali-insoluble complex with chitin and chitosan polymers. Data presented here show that M. restricta has a polysaccharide organization very different of all fungal species analyzed to date.     

Molecular Analysis of Malassezia Microflora from Patients with Pityriasis Versicolor

Background: Pityriasis versicolor (PV) is a superficial infection of the stratum corneum caused by Malassezia species. Eleven species have been identified within this genus, namely M. globosa, M. restricta, M. sympodialis, M. furfur, M. obtusa, M. slooffiae, M. pachydermatis, M. dermatis, M. japonica, M. yamatoensis, M. nana. M. furfur has long been identified as the causative fungus of PV. However, recent studies using the culture and isolation identified by morphological and physiological characteristics suggest that M. globosa is the causative agent of PV. Objectives: The aim of this study was to examine the distribution of PV microorganisms with a molecular-based non-culture method. Patients: The subjects were 49 patients with PV (32 males, 17 females; 16–83 years old) who visited our outpatient clinic. Methods: Samples were taken from lesions for direct microscopy with methylene blue and detected Malassezia species without M. pachydermatis and M. nana using a non-culture-based method consisting of nested PCR with specific primers. Results: The most frequently isolated species were M. globosa and M. restricta (both 93.9%). Only M. globosa was detected from the lesion in which the mycelial form alone was observed microscopically, but M. restricta was not. Conclusions: Our results suggest that M. globosa is the causative agent of PV.     

Prevalencia de especies de Malassezia asociadas a lesiones de dermatitis seborreica en pacientes de Argentina

BackgroundSeborrheic dermatitis (SD) is considered the second most frequently dermatosis associated with the genus Malassezia but little is the knowledge about the epidemiology of this association.AimsTo determinate the prevalence of Malassezia species associated with SD and to analyse their distribution according to the location of the lesion on the body.MethodsThis study was performed in Resistencia city, located in a subtropical area in northeast Argentina. In this study, 226 skin samples from patients with lesions compatible with SD were studied. Age, gender and body sites lesion were recorded. Strains were identified by PCR-RFLP.ResultsOne hundred and thirty-one positive cultures were obtained. Association of 2 species was detected in 10 cases; therefore, 141 strains were isolated. Malasezzia globosa (43.3%) was the most frequent species isolated, followed by Malasezzia furfur (20.6%), Malasezzia sympodialis (17%) and Malasezzia restricta (16.3%). Three isolates of Malasezzia slooffiae (2.1%) and one of Malasezzia pachydermatis (0.7%) were obtained. Statistical significance (P < 0.05) was found between M. globosa and scalp. Malasezzia restricta was isolated only in head areas.ConclusionsThis study suggests M. globosa is the most related species to SD. The prevalence of other species is different from that reported by other authors. Only M. globosa and M. restricta presented a pattern of relationship with the body sites of the lesions. It is noteworthy is the isolation of the zoophylic species M. pachydermatis. The Malassezia genus ecology and the pathogenic role of its species are still under study. This work is a contribution to this knowledge.     

Genotype Analyses of Human Commensal Scalp Fungi,Malassezia globosa,and Malassezia restricta on the Scalps of Patients with Dandruff and Healthy Subjects

Dandruff and seborrheic dermatitis are common afflictions of the human scalp caused by commensal scalp fungi belonging to the genus Malassezia. Malassezia globosa and Malassezia restricta are the predominant species found on the scalp. The intergenic spacer regions of these species’ rRNA genes contain short sequence repeats (SSR): (GT)n and (CT)n in M. globosa and (CT)n and (AT)n in M. restricta. In the present study, we compared the genotypes (SSR) of M. globosa and M. restricta colonizing the scalps of patients with dandruff and healthy individuals. For M. globosa, the genotype (GT)₁₀:(CT)₈ (40.3 %, 25/62) was predominant followed by (GT)₉:(CT)₈ (14.5 %, 9/62) and (GT)₁₁:(CT)₈ (14.5 %, 9/62) in patients with dandruff, whereas the genotypes in healthy subjects were diverse. For M. restricta, the genotype (CT)₆:(AT)₆ (59.7 %, 37/62) was predominant followed by (CT)₆:(AT)₈ (24.2 %, 15/62) in patients with dandruff, while four genotypes, (CT)₆:(AT)₆ (10.5 %, 6/57), (CT)₆:(AT)₇ (22.8 %, 13/57), (CT)₆:(AT)₈ (17.5 %, 10/57), and (CT)₆:(AT)₁₀ (21.1 %, 12/57), accounted for 71.9 % of all combinations in healthy subjects. The results of this study suggest that the M. globosa genotype (GT)₁₀:(CT)₈ and the M. restricta genotype (CT)₆:(AT)₆ may be involved in the development of dandruff.     

Microreview of Pityriasis versicolor and Malassezia species

Recently 11 Malassezia species were isolated. Attention has focused on the relationship between Malassezia species and Malassezia-related disease. The causal fungus of Pityriasis versicolor is M. globosa. The conditions of mycelial form induction are not clear for M. globosa.     

Species of Malassezia associated with various dermatoses and healthy skin in the Mexican population

At the present, eight Malassezia species have been described and their distribution in normal skin and in several skin diseases appears variable. The aim of the present study was to determine the frequency and distribution of Malassezia species in patients with psoriasis, seborrhoeic dermatitis and pityriasis versicolor attended in a Hospital from Mexico City, in addition to a healthy individual group. Scales of abnormal and healthy skin were grown in modified Dixon agar and the species identification was performed by macroscopic and microscopic features; by catalase and urease reaction; growth at 32, 37 and 40 degrees C; and Tween 20, 40, 60 and 80 assimilation. The cultures from 63 persons were included: forty six patients (20 psoriasis, 15 seborrhoeic dermatitis, 11 pityriasis versicolor) and 17 healthy individuals (external auditory canal). A total of 96 isolates were obtained. The more frequently isolated species were: M. sympodialis (38.2%) and M. furfur (26.5%) in psoriasis; M. sympodialis (38.5%) and M. slooffiae (34.6%) in seborrhoeic dermatitis; M. globosa (46.7%) and M. sympodialis (26.7%) in pityriasis versicolor; and M. restricta (47.6%) and M. globosa (23.8%) in normal skin. The number of isolates, the species diversity and association were higher in the patients group than in the healthy individuals group.     

The Role of L-DOPA on Melanization and Mycelial Production in Malassezia Furfur

Melanins are synthesized by organisms of all biological kingdoms and comprise a heterogeneous class of natural pigments. Certain of these polymers have been implicated in the pathogenesis of several important human fungal pathogens. This study investigated whether the fungal skin pathogen Malassezia furfur produces melanin or melanin-like compounds. A melanin-binding monoclonal antibody (MAb) labelled in vitro cultivated yeast cells of M. furfur. In addition, melanization of Malassezia yeasts and hyphae was detected by anti-melanin MAb in scrapings from patients with pityriasis versicolor. Treatment of Malassezia yeasts with proteolytic enzymes, denaturant and concentrated hot acid yielded dark particles and electron spin resonance spectroscopy revealed that these particles contained a stable free radical compound, consistent with their identification as melanins. Malassezia yeasts required phenolic compounds, such as L-DOPA, in order to synthesize melanin. L-DOPA also triggered hyphal formation in vitro when combined with kojic acid, a tyrosinase inhibitor, in a dose-dependent manner. In this respect, L-DOPA is thought to be an essential substance that is linked to both melanization and yeast-mycelial transformation in M. furfur. In summary, M. furfur can produce melanin or melanin-like compounds in vitro and in vivo, and the DOPA melanin pathway is involved in cell wall melanization.     

Identification of Malassezia species isolated from patients with extensive forms of pityriasis versicolor in Siena,Italy

BackgroundPityriasis versicolor (PV) is an infection caused by various species of Malassezia yeast. There is no agreement in the literature concerning the species of Malassezia and the demographic, clinical, and mycological data.AimsTo prospectively identify Malassezia species isolated from lesions of patients with extensive, long standing and recurrent forms of PV and to estimate the relationship between Malassezia species and the demographic and clinical data of the patients.MethodsAll patients with PV were enrolled over a four-year period. Malassezia species were isolated in cultures and identified by morphological features and physiological tests. In the last 2 years a PCR-based technique was used to confirm the species’ identification.ResultsA total of 74 patients (43 males and 31 females, mean age 39.5 years) were enrolled. Only one species was isolated in 45 patients, and more than one species were identified in the remaining 28 patients (38%). M. globosa was the most frequently isolated (60.3%) species. There was a significant association between the isolation of 2 or more species and the presence of at least one predisposing factor. In the last 29 cases, which were subjected to PCR, there were no differences in the identification of isolated species as compared to traditional methods.ConclusionsThe isolation of more than one species in a single lesion is not infrequent in PV and is related to the presence of one predisposing factor. The isolated species isolated were not influenced by demographic and clinical features. The traditional and more recent (PCR) procedures gave the same results in the isolated species.     

Malassezia Intra-Specific Diversity and Potentially New Species in the Skin Microbiota from Brazilian Healthy Subjects and Seborrheic Dermatitis Patients

Malassezia yeasts are part of the resident cutaneous microbiota, and are also associated with skin diseases such as seborrheic dermatitis (SD). The role these fungi play in skin diseases and why they are pathogenic for only some individuals remain unclear. This study aimed to characterize Malassezia microbiota from different body sites in healthy and SD subjects from Brazil. Scalp and forehead samples from healthy, mild SD and severe SD subjects were collected. Non-scalp lesions from severe SD patients were also sampled. 5.8S rDNA/ITS2 amplicons from Malassezia sp. were analyzed by RFLP and sequencing. Results indicate that Malassezia microbiota did not group according to health condition or body area. Phylogenetic analysis revealed that three groups of sequences did not cluster together with any formally described species, suggesting that they might belong to potential new species. One of them was found in high proportions in scalp samples. A large variety of Malassezia subtypes were detected, indicating intra-specific diversity. Higher M. globosa proportions were found in non-scalp lesions from severe SD subjects compared with other areas, suggesting closer association of this species with SD lesions from areas other than scalp. Our results show the first panorama of Malassezia microbiota in Brazilian subjects using molecular techniques and provide new perspectives for further studies to elucidate the association between Malassezia microbiota and skin diseases.     

Extracellular enzymatic activity of Malassezia spp. isolates

Extracellular enzymatic activity of different species of Malassezia spp was evaluated. Thirty-three isolates of animal origin (dogs and cats) and stock culture samples were studied. Twenty isolates of M. pachydermatis, 8 of M. furfur, 2 of M. sympodialis and M. globosa and one of M. restricta, M. obtusa and M. slooffiae were examined. The enzymatic activity was investigatedusing Api Zym system. The enzymatic patterns showed light differences. Esterase lipase, Phosphatase acid and Naphtol-AS-BI-phosphohydrolase were produced in significant amounts from most isolates excepted for M. restricta, confirming the limited enzymatic activity of this species. Data obtained from the other new species described after the revision of the genus, appear to be quite homogeneous. Dixon’s broth appeared to be a valid medium for the growth of all Malassezia spp. This revised version was published online in June 2006 with corrections to the Cover Date.     

Genotyping by RAPD-PCR analyses of Malassezia furfur strains from pityriasis versicolor and seborrhoeic dermatitis patients

Malassezia furfur is lypophilic yeast commonly associate with dermatological disorders. In the present work, we described the isolation of 47 M. furfur strains from three groups of patients: pityriasis versicolor (21 isolates), seborrhoeic dermatitis (15 isolates) and seborrhoeic dermatitis of the HIV positive patients (11 isolates). To investigate the identity of the strains at molecular level, DNA genomic of M. furfur strains were prepared and used to RAPD-PCR analyses. RAPD assay were carried out using two decamer primers and bands pattern generated were analyzed by an Unweighted Pair-Group Method (UPGMA). Dendrogram established a distinct differentiation between M. furfur isolates from pityriasis versicolor and seborrhoeic dermatitis patients with or without AIDS. We concluded that RAPD typing presented a high discriminatory power between strains studied in this work and can be applied in epidemiological investigation of skin disease causing by M. furfur.     

Pityriasis Versicolor: Clinical Spectrum and Diagnosis

Pityriasis versicolor (PV) is caused by the fungus Malassezia, especially by Malassezia globosa. The predisposing factors for PV are heat, humidity, hyperhidrosis, oral contraceptives, stress, application of oily preparations, and treatment with corticosteroids. PV is found mainly on the seborrheic areas of the trunk, shoulders, upper aspects of the arms, and neck, but it may spread to the face, scalp, submammary areas, axillae, groin, skin folds, and buttocks. It is characterized by hyperpigmented, hypopigmented, or erythematous (versicolor), round-to-oval, finely scaling, thin plaques. Other less frequent clinical variants are papular, atrophic, imbricata, and pityriasis rubra pilaris-like presentations. The diagnosis of PV is usually made clinically with the aid of Wood’s light or dermoscopy. Direct microscopic examination, culture, biopsy, and molecular studies are among the laboratory diagnostic methods. Topical therapy is the treatment of choice for most patients. Systemic imidazole therapy is usually reserved for widespread or resistant cases.     

In vitro antifungal efficacy of ciclopirox olamine alone and associated with zinc pyrithione compared to ketoconazole against Malassezia globosa and Malassezia restricta reference strains

The aim of this study was to determine the in vitro fungicidal and growth inhibitory activity of ciclopirox olamine alone (1% and 1.5%) or in association with 1% zinc pyrithione compared to 2% ketoconazole, against Malassezia species particularly involved in the pathogenesis of seborrheic dermatitis. Experiments were performed on Malassezia globosa IP 2387.96 and M. restricta IP 2392.96 strains. Growth inhibitory activity of the active compounds in solution was evaluated by measuring minimal inhibitory concentrations using a broth micro-method and their fungicidal activity by a filtration method after contact times between solutions and yeasts ranging from 3–5 to 30 min. Concerning the determination of minimal inhibitory concentration of ciclopirox olamine/zinc pyrithione, it revealed the marked synergistic inhibitory effect of the association, leading to a higher efficacy compared to ketoconazole. As to the fungicidal activity of ciclopirox olamine, it significantly increased with the contact time. After 15–30 min of contact between 1.5% ciclopirox olamine and Malassezia strains, a 2-log reduction of Malassezia counts was observed. The 1.5% ciclopirox olamine/1% zinc pyrithione association was characterized by a steady fungicidal efficacy whereas the 2% ketoconazole solution did not express any fungicidal effect. In conclusion, this study demonstrates the in vitro inhibitory and fungicidal efficacy of the ciclopirox olamine/zinc pyrithione association against Malassezia species and underscores its potential interest in the treatment of seborrheic dermatitis.     

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。     

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.     

Skin Colonization by Malassezia spp. in Hospitalized Neonates and Infants in a Tertiary Care Centre in North India

Malassezia, a skin colonizer, is associated with multiple skin disorders in adults, and cephalic pustulosis and folliculitis in children. It can cause fungemia in infants and neonates. The time and pattern of colonization, risk factors associated with colonization and causing fungemia in children, are not well understood. The prospective cohort study was conducted to determine the rate of Malassezia species colonization and associated factors in hospitalized neonates and infants. Consecutive 50 neonates and infants admitted in neonatal and pediatric intensive care units were studied. The skin swabs were collected on the day of admission and every fifth day, thereafter, till the patient was discharged or died. Putative risk factors for the colonization of Malassezia species were recorded. Isolates were identified by phenotypic methods and sequencing of the D1 and D2 region of rDNA. Neonates were not colonized at the time of entry in neonatal ICU or at birth. Nineteen (38 %) neonates were colonized with Malassezia species during their hospital stay. Among the infants, three (6 %) came to ICU with Malassezia colonization and 26 (52 %) acquired Malassezia during ICU stay. Mechanical ventilation, duration of hospital stay, central venous catheterization, and antifungal therapy were the significantly associated factors for colonization. Malassezia furfur was the most common species isolated from the skin of infants and neonates. Colonization by Malassezia species in infants and neonates in a hospital is not uncommon and can be a potential source of nosocomial infection.