Seborrheic dermatitis: predisposing factors and ITS2 secondary structure for Malassezia phylogenic analysis.

Seborrheic dermatitis (SD) is a chronic, widespread skin condition, which is considered a multifactorial disease influenced, in part, by Malassezia spp. opportunistic activities, as well as various endogenous and exogenous factors. Malassezia species are lipophilic, lipid-dependent yeasts that are members of the normal mycobiota of the human skin. Their isolation from SD lesions varies around the world and the study of the relationship among factors such as gender, age, immunosuppressive condition of the patient and SD development, can lead to a better understanding of this disease. To elucidate the association of age and gender with the development of SD and to precisely determine the Malassezia species involved in the disease, samples were obtained from 134 individuals, including individuals without lesions, human immunodeficiency virus positive patients, individuals with seborrheic dermatitis, and HIV patients with seborrheic dermatitis. Malassezia spp. were identified by phenotypic and genotypic methods and a phylogenetic analysis was performed using Bayesian inference. This study revealed that age and gender are not predisposing factors for SD development, and that the most frequent species of Malassezia related to SD development among the Colombian population is M. restricta. We also report the isolation of M. yamatoensis for the first time in Colombia, and propose an ITS2 secondary structure from Malassezia taxa that can be used for precise identification and to establish more robust phylogenetic relationships.

Isolation and characterization of two strains of Fusarium oxysporum causing potato dry rot in Solanum tuberosum in Colombia.

BACKGROUND: Fusarium oxysporum has worldwide distribution and causes severe vascular wilt or root rot in many plants. Strains are classified into formae speciales based on their high degree of host specificity, of which multilocus sequence typing provides a fairly good estimate. AIMS: The main aim of this study was to identify the causal agent of an infected potato tuber in Colombia. METHODS: Two F. oxysporum isolates were recovered from a potato tuber showing symptoms of dry rot. Both macroscopic and microscopic morphology differences were observed between the two isolates. Koch's postulates were verified and in quantitative tuber pathogenecity trials, both isolates induced moderate dry rot. Ribosomal internal transcribed spacer (ITS) and partial intergenic spacer region (IGS) sequences were PCR-amplified, sequenced and shown to be identical for the two isolates. A maximum parsimony phylogeny was created using F. oxysporum IGS sequences available in the Genebank database, which does not include sequences from the formae speciales tuberosi. RESULTS: Our two isolates were most closely related to a red clover (Trifolium pratense) pathogenic isolate and two non-pathogenic F. oxysporum isolates from birdsfoot trefoil (Lotus corniculatus) and Lycopersicon sp. rhyzosphere (99% identity). CONCLUSIONS: These experiments showed that our isolates are not restricted to potato and that a molecular marker is needed to differentiate the formae speciales since the IGS and EF-1α do not have the power to do it.

Fusarium species detected in onychomycosis in Colombia.

Fusarium spp. have frequently been isolated from patients with onychomycosis. In Colombia, several studies have shown that Fusarium is the most common non-dermatophyte mould causing onychomycosis and its spread has increased in the past years. In this study, samples were collected in 2003 and 2004 from 137 patients who were diagnosed with onychomycosis caused by Fusarium spp. Three species of Fusarium were identified: Fusarium solani (64.9%), Fusarium oxysporum (32.8%) and Fusarium verticillioides (2.3%). The diseases were more common in women (73%) than in men (27%) and occurred mainly among adults between 31 and 40 years old. The percentage of patients who had received previous treatments was 63.5%. In the last years, new and improved antifungal agents like echinocandins or new triazoles like voriconazole have been developed. For this reason, susceptibility testing using voriconazole was performed, by broth microdilution and disk diffusion. The results showed that F. solani had the highest minimum inhibitory concentration. Using the disk diffusion test, many of the isolates showed variable susceptibility. Genetic diversity of F. oxysporum isolates was determined by random amplified polymorphic DNA. Twenty isolates belonging to different haplotypes were selected for PCR amplification of a region of the gene encoding α-l-arabinofuranosidase B, a specific test to determine if the isolates were F. oxysporum f. sp. dianthi. On the basis of these PCR results, we found that five out of the 20 F. oxysporum isolates corresponded to f. sp. dianthi.

Polimorfismos genéticos de aislamientos del género Malassezia obtenidos en Colombia de pacientes con lesión dermatológica y sin ella / Genetic polymorphism of Malassezia spp. yeast isolates from individuals with and without

Introducción. Las especies del género Malassezia se consideran levaduras oportunistas emergentes de gran importancia. Han sido asociadas a diferentes patologías dermatológicas y sistémicas de las cuales se aislan una o más especies de este género. El papel de estas levaduras en las enfermedades dermatológicas no se ha aclarado completamente, ya que la Malassezia spp. pertenece a la flora normal de la piel. Objetivo. Buscar marcadores genéticos en los aislamientos de Malassezia spp. que permitan correlacionar las lesiones dermatológicas con las especies aisladas. Materiales y métodos. Se obtuvieron 103 aislamientos de Malassezia spp. a partir de muestras de pacientes con pitiriasis versicolor, dermatitis seborreica, dermatitis seborreica en pacientes positivos para VIH, dermatitis atópica, y de individuos sanos. Para los controles se usaron ocho cepas del Centraalbureau voor Schimmelcultures (CBS), Holanda. El perfil genético se realizó utilizando la técnica de ADN polimórfico amplificado aleatorio (RAPD) con tres iniciadores (OPA2, OPA4, OPA13). Los datos obtenidos se analizaron con los programas Diversity Database y SYN-TAX-PC. Resultados. Se observó heterogeneidad genética intraespecífica en Malassezia furfur, Malassezia globosa, Malassezia restricta, Malassezia slooffiae y Malassezia obtusa, mientras que Malassezia sympodialis mostró mayor homogeneidad. Conclusión. No se determinó ningún patrón genético específico mediante la técnica de RAPD para las especies de Malassezia que se pudiera relacionar con la entidad dermatológica implicada

[Endophytic fungi in rose (Rosa hybrida) in Bogota, Colombia]. / Aislamiento de hongos endofitos en rosa (Rosa hybrida) en Bogotá,Colombia.

We have investigated the presence of endophytic fungi associated with rose plants (Rosa hybrida) in Colombia. Endophytic fungi were isolated from healthy leaves of ten ornamental roses plants from gardens cultured in malt extract, peptone, yeast extract agar plates (MPY). We sampled 560 leaves fragments, 56 per sample. Endophytic fungi comprised 92 isolates (16.4%); of these isolates, 41 were classified as sterile mycelium (without reproductive structures that allowed their identification), 31 isolates were identified to genus or to species, and 20 isolates could not be identified at all. The identified endophytic fungi were as follow: Nigrospora oryzae, Aureobasidium spp, Acremonium spp. The fungi Nodulisporium sp, Gliocladium virens, Cladosporium sp, Alternaria sp, Phoma sp and Chaetomium globosum were represented by one isolate each. Since the endophytic fungi are known for their capacity to produce metabolites with biological activity, it is possible that the microorganisms found in this study have potential as antagonist of rose pathogens.

Malassezia yeast species isolated from patients with dermatologic lesions.

INTRODUCTION: Yeasts of the genus Malassezia form a normal component of skin flora, but are also associated with several dermatological disorders. Since 1996, the description of new species in this genus have led to new questions about their epidemiology and pathogenicity. OBJECTIVE: Herein, the frequency of Malassezia species in individuals with pityriasis versicolor, atopic dermatitis, seborrhoeic dermatitis, seborrhoeic dermatitis was compared in HIV patients and healthy individuals. Three body sites were selected for examination--head, thorax, and upper and lower extremities. MATERIAL AND METHODS: The 154 Malassezia species were isolated from 112 individuals and grouped as follows: 39 with seborrhoeic dermatitis (20 were HIV-positive patients), 18 with pityriasis versicolor, 18 with atopic dermatitis and 37 without dermatological leisions. HIV patient samples were examined microscopically, and specimens from both patients and healthy subjects were cultured on modified Dixon agar medium. Subsequently, isolates were identified by macroscopic, microscopic and physiological characteristics. RESULTS: The most commonly isolated species were Malassezia globosa (37.5%), M. sympodialis (31.3%) and M. furfur (31.3%). Malazzerzia globosa predominated in patients with pityriasis versicolor (67%) and in HIV-positive patients with seborrhoeic dermatitis (85%). In non-HIV patients with atopic dermatitis or seborrhoeic dermatitis, M. furfur and M. restricta were isolated in 72% and 26% of the cases, respectively. CONCLUSION: Several conclusions were evident. First, Malassezia species was present in subjects with and without dermatological pathologies. Second, the species frequency in the sampled population differed from frequencies reported from other geographic areas. Third, Malassezia globosa was involved at high frequency in patients with dermatological pathologies, suggesting a higher pathogenicity of this species. Additional studies on each species are recommended to clarify their pathogenic roles in association with HIV-positive and normal subjects.

Malassezia yeast species isolated from patients with dermatologic lesions / Especies de Malassezia aisladas de pacientes con lesiones dermatológicas

Introducción. Las levaduras del género Malassezia forman parte de la flora normal de la piel y se asocian con varios tipos de lesiones dermatológicas. Desde 1996, la descripción de nuevas especies dentro de este género planteó numerosos interrogantes sobre su epidemiología y patogenicidad. Objetivo. El fin de este trabajo fue encontrar la frecuencia de las diferentes especies de Malassezia en individuos con pitiriasis versicolor, dermatitis atópica, dermatitis seborreica, dermatitis seborreica en pacientes positivos para VIH, y en individuos sin lesiones a partir de tres zonas corporales (cabeza, tórax, extremidades inferiores y superiores). Materiales y métodos. Se identificaron 154 aislamientos de especies de Malassezia de 112 individuos: 39 con dermatitis seborreica de los cuales 20 eran positivos para VIH, 18 con pitiriasis versicolor, 18 con dermatitis atópica y 37 individuos sin lesión dermatológica (controles). Las escamas de piel de los pacientes se observaron microscópicamente y las muestras de pacientes y controles se cultivaron en agar Dixon modificado. Posteriormente, se observaron las colonias y se identificaron según sus características macroscópicas, microscópicas y fisiológicas. Resultados. Las especies más frecuentemente aisladas en el total de los grupos estudiados fueron: Malassezia globosa (37,5 por ciento), M. sympodialis (31,3 por ciento) y M. furfur (31,3 por ciento). M. globosa predominó en los aislamientos de pacientes con pitiriasis versicolor (67 por ciento) y en pacientes positivos para VIH con dermatitis seborreica (85 por ciento). En pacientes con dermatitis atópica y dermatitis seborreica sin diagnóstico de VIH, se aislaron M. furfur y M. restricta en el 72 por ciento y el 26 por ciento de los casos, respectivamente. Conclusiones. Las diferentes especies de Malassezia pueden aislarse de pacientes con lesiones dermatológicas o sin ellas. Además, la frecuencia de especies en la población muestreada difiere de lo reportado en otras zonas geográficas. Se destaca la alta frecuencia de M. globosa en pacientes con lesiones lo que podría indicar la mayor patogenicidad de esta especie. No obstante, se requieren estudios adicionales para determinar el papel de las diferentes especies del género Malassezia, asociadas con estas patologías

[Genetic polymorphism of Malassezia spp. yeast isolates from individuals with and without dermatological lesions]. / Polimorfismos genéticos de aislamientos del género Malassezia obtenidos en Colombia de pacientes con lesión dermatológica y sin ella.

INTRODUCTION: Malassezia spp. yeasts are opportunistic and newly emergent diseases. One or more species have been isolated in association with dermatological pathology and systemic disease. Their pathological role has not been fully elucidated since Malassezia spp. are common in normal skin flora. OBJECTIVE: In the current study, a search was undertaken for genetic markers in the Malassezia spp. isolates that correlate with each type of dermatologic lesion. MATERIALS AND METHODS: A total of 103 strains of Malassezia spp were isolated from patients with pytiriasis versicolor, seborrheic dermatitis, seborrheic dermatitis HIV (+), and atopic dermatitis, as well as from healthy individuals. Eight isolates from CBS Centraalbureau voor Schimmelcultures (Netherlands) were used as controls. Fingerprinting was done using random amplification of polymorphic DNA technique (RAPD) with three primers (OPA2, OPA4, OPA13). The data were analyzed with Diversity Database and SYN-TAX-PC programs. RESULTS: Intraspecies genetic heterogeneity in M. furfur, M. globosa, M. restricta, M. slooffiae, M. obtusa was observed, whereas M. sympodialis showed the greatest homogeneity. CONCLUSION: The dermatological disease caused by these different species was not associated with distinctive RAPD fingerprints.