Aislamiento e identificación fenotípica y genotípica de cepas de Malassezia spp. en pacientes con pitiriasis versicolor / Isolation and identification phenotypic and genotypic strains of Malassezia spp. in patients with pytiriasis versicolor

Antecedentes: Pitiriasis versicolor (PV) es una infección micótica superficial crónica de la piel, debido a la colonización del estrato córneo por Malassezia sp. Tradicionalmente M. furfur era considerada el agente causal de PV, hoy en día se ha demostrado que las especies de Malassezia más comunes cultivadas a partir de de lesiones de PV son M. globosa y M. sympodialis. Objetivos: Evaluar la epidemiología de la dermatomicosis PV, en pacientes de la ciudad de Talca, Chile. Metodología: En este estudio, 19 pacientes diagnosticados con PV fueron evaluados. Se realizó identificación fenotípica e identificación molecular por técnica de PCR-RFLP. Para esto, se diseñaron partidores que amplificaron una zona del 26S ADNr presente en todas las especies de Malassezia. Resultados: De un total de 19 pacientes con PV, 7 correspondieron a sexo masculino (36,8 por ciento) y 12 a sexo femenino (63,2 por ciento). Se obtuvieron 18 cepas, a partir de las pruebas fenotípicas fue posible identificar presuntivamente 11 especies de M. globosa (61,1 por ciento) y 7 especies de M. sympodialis (38,9 por ciento). Con los resultados obtenidos en la PCR-RFLP fue posible identificar 11 especies de M. globosa (61,1 por ciento) y 7 especies (38,9 por ciento) de M. sympodialis. Conclusiones: La especie identificada fenotípicamente y molecularmente en mayor proporción fue Malassezia globosa seguida de Malassezia sympodialis. Por lo que hay concordancia entre ambas técnicas de identificación. Este es el primer trabajo de investigación realizado en Talca, Chile, que identificó las especies de Malassezia spp. involucradas en PV, siendo un aporte al estudio de este cuadro clínico.

Background: Pytiriasis versicolor (PV) is a fungal infection of the skin due to the stratum corneum colonization by Malassezia sp. M. furfur traditionally was considered the etiological agent of PV but today it has been shown that M. globosa and M. sympodialis are the most common Malassezia species cultivated from PV lesions. Objective: To evaluate the epidemiological features of PV in patient of the Talca city, Chile. Method: In this study, 19 patients with diagnosis of PV were evaluated. Phenotypic and molecular identified were performed using PCR-RFLP. To do this, primers to amplify 26S ADNr secuence were designed. Result: From a total of 19 patients who presented PV, 7 were male (36.8 percent) and 12 female patients (63.2 percent). Eighteen strains were obtained, from the phenotypic tests, it was possible to presumably identify 11 species of M. globosa (61.1 percent) and 7 species of M. sympodialis (38.9 percent). With PCRRFLP results it was possible to identify 11 species of M. globosa (61.1 percent) and 7 species (38.9 percent) of M. sympodialis. Conclusion: The most commonly isolated species were Malassezia globosa followed by Malassezia sympodialis. So, there is agreement between both techniques of identification. This is the first research work carried out in Talca, Chile, which identified the species of Malassezia spp. Involved in PV, being a contribution to the clinical diagnosis.

Is Malassezia nana the main species in horses' ear canal microbiome?

ABSTRACT The objective of this study was to characterize genotypically Malassezia spp. isolated from the external ear canal of healthy horses. Fifty-five horses, 39 (70.9%) males and 16 (29.1%) females, from different breeds and adults were studied. External ear canals were cleaned and a sterile cotton swab was introduced to collect cerumen. A total of 110 samples were cultured into Dixon medium and were incubated at 32 °C for up to 15 days. Macro- and micromorphology and phenotypic identification were performed. DNA was extracted, strains were submitted to polymerase chain reaction technique, and the products obtained were submitted to Restriction Fragment Length Polymorphism using the restriction enzymes BstCI and HhaI. Strains were sent off to genetic sequencing of the regions 26S rDNA D1/D2 and ITS1-5.8S-ITS2 rDNA. Malassezia spp. were isolated from 33/55 (60%) animals and 52/110 (47%) ear canals. No growth on Sabouraud dextrose agar was observed, confirming the lipid dependence of all strains. Polymerase chain reaction-Restriction fragment length polymorphism permitted the molecular identification of Malassezia nana - 42/52 (81%) and Malassezia slooffiae - 10/52 (19%). Sequencing confirmed RFLP identification. It was surprising that M. nana represented over 80% of the strains and no Malassezia equina was isolated in this study, differing from what was expected.

Identificación de especies de Malassezia aisladas de piel sana en pobladores de Lima, Perú / Identification of Malassezia species isolated from healthy skin in residents of Lima, Peru

Objetivo: Identificar las especies de Malassezia en zonas seborreicas de piel sana en población limeña. Diseño: Estudio descriptivo transversal. Lugar: Instituto de Medicina Tropical æDaniel Alcides CarriónÆ, Universidad Nacional Mayor de San Marcos, Lima, Perú. Participantes: Pobladores asintomáticos. Intervenciones: Empleando la técnica de Mariat y Adan-Campos, se recolectó muestras de piel de 129 pobladores asintomáticos de diversos distritos de la ciudad de Lima. El aislamiento de Malassezia se realizó en medio Dixon modificado e incubado a 32 grados centígrados por 7 días, la identificación de las colonias por sus características macro y micromorfológicas, y la tipificación mediante el estudio de las propiedades bioquímicas y fisiológicas según la técnica de Guillot y col. Principales medidas de resultados: Especie de Malassezia, sexo, edad y región anatómica. Resultados: Se aisló Malassezia spp en 43,4 por ciento de los pobladores, obteniéndose 49,2 por ciento en varones y 37,5 por ciento en mujeres. De las diferentes regiones corporales, 68 cultivos fueron positivos: cuero cabelludo 31 (45,6 por ciento), espalda 36 (52,9 por ciento) y región frontal 1 (1,5 por ciento). El grupo etario con mayor frecuencia de aislamientos (47,2 por ciento) fue el de 14 a 25 años (adolescentes jóvenes). M. slooffiae fue encontrado en 83,8 por ciento y M. obtusa en 16,2 por ciento de los casos. Conclusiones: Se encontró Malassezia spp. en la piel humana sana. M. slooffiae fue la especie predominante de los casos positivos (83,8 por ciento) seguido de M. obtusa (16,2 por ciento)...

Objective: To identify Malassezia species in healthy skin seborrhea areas in Lima inhabitants. Design: Cross-sectional study. Setting: Daniel Alcides Carrion Tropical Medicine Institute, Universidad Nacional Mayor de San Marcos, Lima, Peru. Participants: Asymptomatic persons. Interventions: Skin samples were collected from 129 asymptomatic residents of several districts of Lima city using Mariat and Adan-CamposÆ technique. Malassezia isolation was performed in modified Dixon medium and incubated at 32 grades centigrades for 7 days. Colonies were identified by macro and micro morphological characteristics and typing was determined by biochemical and physiological properties using GuillotÆs technique. Main outcome measures: Malassezia species, participantsÆ gender, age and anatomical region. Results: Malassezia spp was isolated in 43.4 per cent of the residents, 49.2 per cent in men and 37.5 per cent in women. From various body regions 68 cultures were positive: scalp 31 (45.6 per cent), back 36 (52.9 per cent) and frontal region 1 (1.5 per cent). Isolates most common age group (47.2 per cent) was that of adolescents-young (14-25 year-old). M. slooffiae was found in 83.8 per cent and M. obtusa in 16.2 per cent of cases. Conclusions: Malassezia spp. was present in healthy human skin. M. slooffiae was the predominant species in positive cases (83.8 per cent) followed by M. obtusa (16.2 per cent)...

Dermatite seborreica / Seborrheic dermatitis

A dermatite seborreica é uma doença eritêmato-escamativa de caráter crônico-recidivante que acomete entre 1 e 3 por cento da população geral dos Estados Unidos. Possui dois picos de incidência - o primeiro, durante os três primeiros meses de vida, e o segundo, a partir da puberdade, atingindo seu ápice entre os 40 e 60 anos de idade. Os indivíduos HIV positivos têm maior prevalência da doença, que apresenta maior intensidade e tendência à refratariedade ao tratamento. Doenças neurológicas e outras doenças crônicas também estão associadas ao desenvolvimento da dermatite seborreica. Como mecanismo fisiopatogênico, reconhece-se que o fungo Malassezia sp., presente na pele de indivíduos suscetíveis, leve a uma irritação não-imunogênica a partir da produção de metabólitos à base de ácidos graxos insaturados deixados na superfície cutânea. Este artigo faz uma revisão da literatura sobre dermatite seborreica, com ênfase nos aspectos imunogenéticos, formas clínicas e tratamento.

Seborrheic dermatitis is a chronic relapsing erythematous scaly skin disease, the prevalence of which is around 1 to 3 percent of the general population in the United States. It has two incidence peaks, the first in the first three months of life and the second beginning at puberty and reaching its apex at 40 to 60 years of age. The prevalence of seborrheic dermatitis is higher in HIV-positive individuals and the condition tends to be more intense and refractory to treatment in these patients. Neurological disorders and other chronic diseases are also associated with the onset of seborrheic dermatitis. The currently accepted theory on the pathogenesis of this disease advocates that yeast of Malassezia spp., present on the skin surface of susceptible individuals, leads to a non-immunogenic irritation due to the production of unsaturated fatty acids deposited on the skin surface. This article provides a review of the literature on seborrheic dermatitis, focusing on immunogenetics, the clinical forms of the disease and its treatment.

Novos aspectos na evolução clínica da pitiríase versicolor / New aspects in the clinical course of pityriasis versicolor

FUNDAMENTO: A pitiríase versicolor é uma doença infecciosa causada por várias espécies de Malassezia com uma tendência a se tornar recidivante ou crônica. OBJETIVOS: Este trabalho foi conduzido na tentativa de conhecer a evolução clínica da pitiríase versicolor em relação ao número de recidivas após um tratamento adequado no período de 12 meses e correlacionar o número de recidivas com as espécies de Malassezia isoladas. MATERIAL E MÉTODOS: Cento e dois pacientes com diagnóstico clínico e laboratorial de pitiríase versicolor foram acompanhados por um período de 12 meses para observarmos o número de recidivas da doença. RESULTADOS: A pitiríase versicolor, após um tratamento adequado, apresentou três tipos de evolução clínica num período de 12 meses: pitiríase versicolor sem nenhum episódio de recidiva (32,35 por cento); pitiríase versicolor recidivante, com um a quatro episódios de recidiva (52,94 por cento) devidos a fatores de predisposição relacionados; e pitiríase versicolor crônica, com mais de quatro episódios de recidiva (14,70 por cento) sem nenhuma relação com fatores de predisposição. CONCLUSÕES: A pitiríase versicolor apresentou uma evolução clínica de acordo com o número de episódios de recidiva da doença analisados durante um período de 12 meses que pode ser considerada da seguinte maneira: pitiríase versicolor com cura clínica e micológica, pitiríase versicolor recidivante e pitiríase versicolor crônica.

BACKGROUND: Pytiriasis versicolor is an infectious disease caused by several Malassezia species which has a tendency to become relapsing or chronic. OBJECTIVES: This study was conducted in an attempt to investigate the clinical course of pityriasis versicolor with regard to the number of relapses after a 12-month therapy and correlate this number with isolates of Malassezia species. MATERIAL AND METHODS: 102 patients with clinical and laboratory diagnosis of pityriasis versicolor were monitored for 12 months to investigate the number of relapsing episodes of the disease. RESULTS: After appropriate treatment, pityriasis versicolor presented three types of clinical course: pity - riasis versicolor without relapsing episodes (32.35 percent), relapsing pityriasis versicolor with one to four relapsing episodes (52.94 percent ) due to associated predisposing factors, and chronic pityriasis versicolor with more than four relapsig episodes (14.70 percent) with no relation to predisposing factors. CONCLUSIONS: The clinical course of pityriasis versicolor varied according to the number of relapsing episodes of the disease analyzed over a period of 12 months and can be classified as follows: pityriasis versiolor with clinical and mycological clearing, relapsing pityriasis versicolor and chronic pityriasis versicolor.

Identificação de espécies de malassésia na pitiríase versicolor em um serviço de dermatologia do sul do Brasil / Identification of Malassezia yeast species isolated from patients with pityriasis versicolor

A pitiríase versicolor é uma doença de distribuição universal. Existe a descrição de 12 espécies de malassezia. O objetivo deste estudo foi determinar quais as espécies de malassezia mais prevalentes nos pacientes com pitiríase versicolor. Foram realizadas as coletas através de raspado das lesões nos pacientes com suspeita clínica de pitiríase versicolor e posterior exame micológico e cultural para identificação final da espécie. Foram coletadas amostras de 87 pacientes. Quanto às culturas, 30 percent foram de Malassezia sympodialis, 25,7 percent de Malassezia furfur, 22,7 percent de Malassezia globosa, 12,1 percent de Malassezia retrita, 7,6 percent de Malassezia obtusa e 1,5 percent de Malassezia sloofiae.

Pityriasis versicolor (PV) is a disease with worldwide distribution. Twelve different species of Malassezia yeast have been described. The objective of this study was to determine which species of Malassezia are more prevalent in patients with pityriasis versicolor. Samples were collected by scraping the lesions of 87 patients with a clinical suspicion of pityriasis versicolor. The samples were then submitted to fungal microscopy and culture to identify the species. The species found were: Malassezia sympodialis (30 percent), Malassezia furfur (25.7 percent), Malassezia globosa (22.7 percent), Malassezia restricta (12.1 percent), Malassezia obtusa (7.6 percent) and Malassezia slooffiae (1.5 percent).

Comportamiento de malassezia furfur en medios de cultivo con base en los exudados gomosos de spondias dulcis y spondias mombin: producción de lipasa extracelular / Growth of malassezia furfur in media with spondias dulcis and spondias mombin gum exudates: production of extracellular lipase

El desarrollo de las levaduras del género Malassezia requiere condiciones especiales, estos hongos que producen afecciones en la piel, son generalmente cultivados en el medio Dixon. Se ensayaron los exudados gomosos de Spondias dulcis y Spondias mombin como sustratos para Malassezia furfur. Se evaluó también la producción de lipasa. Se determinó la cinética de crecimiento a un determinado intervalo de tiempo (0-168h), y a diferente concentración (0,5; 1%) y pH (4,5; 6,0). La biomasa obtenida para la levadura probada demostró que los sustratos preparados con los exudados gomosos son adecuados para su desarrollo. La mayor actividad de lipasa extracelular se observó al tiempo inicial de estudio (18h) en ambos sustratos, en las condiciones usadas: concentración (0,5 y 1%) y pH(4,5 y 6,0). Estas especies botánicas, ampliamente localizadas en Venezuela, especialmente en los Estados Zulia y Falcón, producen abundante goma. Este hecho, y los resultados obtenidos podrían ser útiles en la obtención de un nuevo sustrato, que pueda competir con el medio Dixon para el aislamiento y la caracterización de especies de Malassezia, y para la producción de lipasa

The development of genus Malassezia yeasts requires special conditions. This fungus, which produces skin diseases, is generally cultivated in the Dixon medium. Gum exudates from Spondias dulcis and Spondias mombin were tested as substrates for Malassezia furfur. Lipase production was also evaluated. The growth kinetic was determined for a given time range (0-168h) at different concentrations (0.5; 1%) and pH levels (4.5; 6.0). The biomass obtained for the tested yeast showed that substrates prepared with S. dulcis and S.mombin gum exudates are suitable for its development. The highest extracellular lipase activity was observed at 18h on both substrates at given concentrations (0.5; 1%) and pH (4.5; 6.0). These botanical species, widely located in Venezuela, especially in the States of Zulia and Falcon, yield abundant gum. Findings may be useful for obtaining new substrates that could compete with the Dixon medium for isolation and characterization of Malassezia species and for lipase production

Pitiríase versicolor: isolamento e identificação das principais espécies de Malassezia / Pityriasis versicolor: isolation and identification of the main species

As espécies do gênero Malassezia isoladas foram: Malassezia sympodialis (16,66 por cento), Malassezia furfur (12,50 por cento), Malassezia globosa (11,45 por cento) e Malassezia slooffiae (2,10 por cento). A Malassezia sympodialis foi a espécie que predominou em nosso estudo. As espécies de Malassezia identificadas não mostraram correlação com as variantes clínicas e com a distribuição das lesões de pitiríase versicolor quanto às regiões do corpo.

Species of the genus Malassezia isolated were: Malassezia sympodialis (16.66 percent), Malassezia furfur (12.50 percent), Malassezia globosa (11.45 percent), and Malassezia slooffiae (2.10 percent). Malassezia sympodialis predominated in the study. The species of Malassezia identified did not show correlation with clinical variants and with the distribution of pityriasis versicolor lesions in relation to areas of the body.

Actividad lipasa de especies de Malassezia aisladas en pacientes sanos y con lesiones dérmicas / Lipase activity of Malassezia species isolated from healthy patients and having dermic lesions

Las levaduras del género Malassezia forman parte de la micro biota normal de piel humana y animal. Excepto M. pachydermatis, todas las especies de este género son lipodependientes. Bajo ciertos factores, Malassezia se asocia como agente etiológico en diversas afecciones dérmicas. Uno de los principales factores de virulencia de estas levaduras es su actividad de lipasa (AL). El objetivo de este trabajo fue introducir modificaciones a las técnicas de determinación de la actividad lipasa (AL) para su aplicación en levaduras lipodependientes y estudiar la AL en cepas de Malassezia aisladas de personas con piel sana y de pacientes con pitiriasis versicolor (PV), dermatitis seborreica (DS) y psoriasis (PS). Se estudiaron 94 cepas aisladas de 34 pacientes con lesiones de PV, 20 con DS, 7 con PS y 33 cepas de personas con piel sana. Las modificaciones planteadas a la técnica, que incluyeron variación del medio de cultivo y tiempos de incubación, permitieron la determinación semi cuantitativa de la AL con resultados claros y definidos. El 88,23 por ciento de las cepas presentó AL. No se obtuvieron diferencias estadísticamente significativas al comparar la AL entre las cepas de pacientes con afecciones de piel y las cepas aisladas de personas sanas. La producción de lipasas de las especies de Malassezia en orden decreciente fue: M. sympodialis, M. slooffiae, M. furfur, M. globosa, M. restricta. M. globosa y M. furfur fueron las especies en que se observaron mayor cantidad de cepas no productoras de AL y cepas con gran variabilidad en la medida de AL.

Yeasts of the genus Malassezia are part of the regular microbiota in human and animal skin. Except for M. pachydermatis, all the species of this genus are lipodependent. Malassezia, under certain factors, is associated as an etiological agent in diverse dermic affections. One of the main virulence factors of these yeasts is their lipase activity (LA). The objective of this research was to introduce some changes in the techniques adopted to determine the lipase activity (LA) in order to apply them to lipodependent yeasts and to study likewise the LA in Malassezia strains isolated from healthy skin people and patients diagnosed with pitiriasis versicolor (VP), greasy dermatitis (GD) and psoriasis ( PS). Ninety four strains isolated from 34 patients having VP lesions, 20 with GD, 7 with PS and 33 strains from healthy skin people. Changes suggested to the technique involved a variation in the medium of culture as well as in the time of incubation what resulted in the semiquantitative determination of the LA together with clear and precise results. The presence of LA was observed in of 88.23 percent strains. The comparison of the LA among strains of patients bearing injured skin and those isolated from healthy skin did not show any significant statistical difference. The production of lipasae from Malassezia species were in decreasing order: M.sympodialis, M. slooffiae, M.furfur,M.globosa and M.restricta. M. globosa and M. furfur were the species that revealed the highest number of non producting LA strains as well as strains with the highest variability in the degree of LA.

Identificação de espécies de Malassezia em pacientes com pitiríase versicolor em Goiânia-GO / Identification of Malassezia species in patients with pityriasis versicolor in Goiânia-GO

O objetivo deste estudo foi verificar a freqüência de pitiríase versicolor e identificar leveduras do gênero Malassezia, de pacientes encaminhados ao laboratório de Micologia da Universidade Federal de Goiás em Goiânia. Foram diagnosticados 95 casos de pitíriase versicolor e identificados quatro espécies de Malassezia: Malassezia furfur, Malassezia sympodialis, Malassezia globosa e Malassezia obtusa.

The objectives of this study were to investigate the frequency of pityriasis versicolor occurrence and to identify yeasts of the genus Malassezia in patients at the mycology laboratory of the Federal University of Goiás, in Goiânia, State of Goiás. Ninety-five cases of pityriasis versicolor were diagnosed, and four species of Malassezia were identified: Malassezia furfur, Malassezia sympodialis, Malassezia globosa and Malassezia obtusa.

Caracterización de especies del género Malassezia en pacientes con dermitis seborreica y en controles / Characterization of species of the gender Malassezia in patients with seborrheic dermatitis and subjects without skin lesions

The yeast Malassezia spp has an established etiological role in pityriasis versicolor, folliculitis, systemic infections and onychomycosis. Aim: To assess the presence of Malassezia spp in patients with seborrheic dermatitis (SD), to find a correlation between Malassezia spp count and the severity of the disease and to compare the prevalence of the different Malassezia species in SD patients and subjects without skin lesions. Patients and methods: Scrapings of the face from 81 patients with SD (69 males) and 79 subjects (54 males) without skin lesions were obtained for a direct microscope examination and yeast culture. Results: The yeast Malassezia was found in 76% of SD patients and in 82% of subjects without skin lesions. There was a positive correlation between the number of yeasts found on direct examination and the clinical severity of lesions in SD patients. Although this correlation was statistically significant (p=0.046), the degree of association (rho=0.22) was weak. Fifty Malassezia species were identified. M globosa was found in 67% of SD patients, followed by M furfur and M sympodialis, each present in 16.5% of the SD patients. In subjects without skin lesions, the most prevalent species were M globosa (77%), followed by M sympodialis (12%), M slooffiae (7%) and M furfur (4%). Conclusions: The presence of the yeast Malassezia is not associated with the presence of skin lesions (Rev Méd Chile 2003; 131: 1295-300).

Actual taxonomía del género Malassezia / Current taxonomy of Malassezia genus

Desde 1853 a la fecha, las levaduras del género Malassezia han recibido diferentes denominaciones debido a su gran polimorfismo y dificultad para cultivarlas. Se revisa la evolución histórica del género, concluyendo en la taxonomía actual propuesta por Guého, Midgley y Guillot, que agrupa siete especies dentro del género Malassezia.

From 1853 to date, yeasts of the Malassezia genus have received different denominations due to their great polymorphism and cultivation difficulties. We review the historic evolution of the genus, giving way to the current taxonomy proposed by Guého, Midgley and Guillot, which groups them into seven species.