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.     

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.     

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.     

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.     

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.     

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.     

<Emphasis Type="Italic">Malassezia</Emphasis> spp. in Acoustic Meatus of Bats (<Emphasis Type="Italic">Molossus molossus</Emphasis>) of the Amazon Region,Brazil

The yeasts of the Malassezia genus are opportunistic microorganisms and can cause human and animal infections. They are commonly isolated from the skin and auricular canal of mammalians, mainly dogs and cats. The present study was aimed to isolate Malassezia spp. from the acoustic meatus of bats (Molossus molossus) in the Montenegro region, “Rondônia”, Brazil. From a total of 30 bats studied Malassezia spp. were isolated in 24 (80%) animals, the breakdown by species being as follows (one Malassezia sp. per bat, N = 24): 15 (62.5%) M. pachydermatis, 5 (20.8%) M. furfur, 3 (12.5%) M. globosa and 1 (4.2%) M. sympodialis. This study establishes a new host and anatomic place for Malassezia spp., as it presents the first report ever of the isolation of this genus of yeasts in the acoustic meatus of bats.     

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.     

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.     

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.     

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.     

Gentamycin inhibits the growth of Malassezia pachydermatis in culture

BackgroundMalassezia pachydermatis is a yeast of importance in both veterinary and human medicine.AimsTo know if M. pachydermatis grow on micological media with high concentrations of gentamycin.MethodsTwenty M. pachydermatis strains were streaked on Sabouraud Dextrose Agar plates with different concentrations of gentamycin.ResultsAll isolates were inhibited when high concentrations of gentamycin were added.ConclusionsThe use of plates with high concentrations of gentamycin can lead to some important misdiagnoses: firstly, false-negative cultures, and secondly, an erroneous classification of M. pachydermatis as a lipid-dependent species. Morever, all of this could be useful in two therapeutic fields: i) in animals, topical gentamycin could be an efficacious treatment for a disease such as external otitis in dogs; ii) in humans, we hypothesize that gentamycin could be regarded as a possible therapy (“antibiotic-lock”) for catheter-associated Malassezia spp. infections.     

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.     

Producción de fosfolipasa y proteinasa en cepas de Malassezia pachydermatis aisladas de perros con otitis y sin otitis

BackgroundMalassezia pachydermatis is part of the skin microbiota of dogs and cats. M. pachydermatis has been associated with external otitis and seborrhoeic dermatitis, reported more often in dogs than in cats. When the physical, chemical or immunological mechanisms of the skin are altered, M. pachydermatis could act as a pathogen. Thus, several virulence factors, such as the ability to produce esterase, lipase, lipoxygenase, protease, chondroitin sulphatase, and hyaluronidase, have been studied.AimsIn the present study, we aim to identify the phospholipase activity measured at pH 6.3, and the proteinase activity measured at pH 6.3 and pH 6.8 (pH from ears of dogs with external otitis) of M. pachydermatis strains isolated from dogs with and without external otitis.MethodsThe phospholipase activity was measured using a semi-quantitative method with egg yolk, and the proteinase activity with a semi-quantitative method using bovine serum albumin agar. The study was performed on 96 isolates of M. pachydermatis, 43 isolated from dogs without clinical symptoms of otitis, and 52 isolated from dogs with otitis.ResultsIn our study, 75.8% of the isolates showed phospholipase activity at pH 6.3, and 81 and 97.9% of them showed proteinase activity measured at pH 6.3 and 6.8, respectively. A higher phospholipase activity was detected in strains isolated from dogs with otitis. The proteinase activity was increased at a pH of 6.8 (97.9%) in comparison to a pH of 6.3 (81%).ConclusionsOur results suggest that the phospholipase activity may play an important role in the invasion of host tissues in chronic canine otitis cases. The proteinase activity results obtained in this study suggest that a reduction in the pH of the treatment may improve its efficacy in the resolution of M. pachydermatis otitis.     

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 Identification of <Emphasis Type="Italic">Malassezia</Emphasis> Species in Patients with <Emphasis Type="Italic">Malassezia</Emphasis> folliculitis in Sfax,Tunisia

Aim

Malassezia folliculitis is caused by the invasion of hair follicles by large numbers of Malassezia cells. Several Malassezia researches still use cultures, morphology and biochemical techniques. The aim of this study was to identify Malassezia species isolated from patients diagnosed with folliculitis, at the Parasitology and Mycology Laboratory of Sfax University Hospital, and to explore the genetic diversity of Malassezia by using PCR-RFLP and PCR-sequencing targeting the rDNA region of the Malassezia genome.

Patients and Methods

Specimens were taken from 27 patients with Malassezia folliculitis. For the molecular identification, PCR amplification of the 26S rDNAD1/D2 region was carried out using the Malup and Maldown primers and three restriction enzymes (BanI, MspI and HeaII) for RFLP analysis. The nucleotide sequences of each isolate were compared to those in the NCBI GenBank by using BLASTIN algorithm.

Results

Three species of Malassezia yeasts were identified among the 31 Malassezia strains isolated: M. globosa (83.9%), M. sympodialis (12. 9%) and M. furfur (3.2%). The sequence analysis of M. globosa showed six genotypes.

Conclusion

There is a high genotypic variability of M. globosa colonizing patients with folliculitis.

     

Heterologous expression and biochemical characterisation of the recombinant β-carbonic anhydrase (MpaCA) from the warm-blooded vertebrate pathogen malassezia pachydermatis

Warm-blooded animals may have Malassezia pachydermatis on healthy skin, but changes in the skin microenvironment or host defences induce this opportunistic commensal to become pathogenic. Malassezia infections in humans and animals are commonly treated with azole antifungals. Fungistatic treatments, together with their long-term use, contribute to the selection and the establishment of drug-resistant fungi. To counteract this rising problem, researchers must find new antifungal drugs and enhance drug resistance management strategies. Cyclic adenosine monophosphate, adenylyl cyclase, and bicarbonate have been found to promote fungal virulence, adhesion, hydrolase synthesis, and host cell death. The CO₂/HCO₃^-/pH-sensing in fungi is triggered by HCO₃^- produced by metalloenzymes carbonic anhydrases (CAs, EC 4.2.1.1). It has been demonstrated that the growth of M. globosa can be inhibited in vivo by primary sulphonamides, which are the typical CA inhibitors. Here, we report the cloning, purification, and characterisation of the β-CA (MpaCA) from the pathogenic fungus M. pachydermatis, which is homologous to the enzyme encoded in the genome of M. globosa and M. restricta, that are responsible for dandruff and seborrhoeic dermatitis. Fungal CAs could be thus considered a new pharmacological target for combating fungal infections and drug resistance developed by most fungi to the already used drugs.     

A reevaluation of the genus Malassezia by means of genome comparison

Results of a study of the genus Malassezia on the basis of genome characters confirm that two species should be maintained, M. furfur and M. pachydermatis. The two forms associated with skin disease, frequently referred to as Pityrosporum orbiculare and P. ovale, were found to be synonymous, the name M. furfur having priority. Malassezia pachydermatis, hitherto regarded as a strictly zoophilic species, may also be found on humans.     

The diversity ofMalassezia yeasts confirmed by rRNA sequence and nuclear DNA comparisons

One hundred and fourMalassezia strains (52 isolated from humans and 52 from animals) were compared using large subunit (LSU) ribosomal RNA sequence similarity and nuclear DNA complementarity. Eight groups of strains were recognized as genetically distinct species. Each taxon was confirmed by a homogeneous mole % GC and percentages of DNA/DNA reassociations higher than 85%. The non-lipid-dependentMalassezia yeasts were maintained as the unique taxonM. pachydermatis. In contrast, lipid-dependent strains were shown to be distributed among seven species:M. furfur, M. sympodialis andM. species 1–5. These taxa matched remarkably well with morphological and serological differences documented by previous investigators. The LSU rRNA sequences allowed a further intraspecific resolution with most of genomic taxa represented by several closely related sequences:M. pachydermatis counted up to seven sequences,M. furfur four sequences,M. species 1 comprised three sequences andM. species 2 andM. species 5 two sequences. Three species,M. sympodialis, M. species 3 andM. species 4, displayed a unique type of sequence. Thus, the present report demonstrates the usefulness of sequencing for both taxonomic and epidemiological purposes.