RESUMO: Malassezia pachydermatis é uma levedura lipofílica e dependente de lipídios, principalmente da pele de animais. Sendo, por essa razão, considerada uma espécie zoofílica e causadora de otite externa em cães. Neste sentido, aspectos associados à sua epidemiologia e patogenicidade constituem um tema de interesse científico. O objetivo deste estudo foi realizar a caracterização molecular de 43 isolados de M. pachydermatis obtidos a partir de cães com otite externa. Para esta propósito, foram amplificadas, sequenciadas e analisadas com enzimas de restrição as regiões do gene 5.8S, do espaçador interno transcrito 2 (ITS2) e D1/D2 do 26S do rRNA pelo método RFLP, com as endonucleases AluI, CfOI e BstF5I. Análises filogenéticas revelaram que os isolados se agruparam com as sequências tipo I, IV e V de M. pachydermatis como já descrito anteriormente. De maneira interessante, se observou um novo RFLP polimórfico utilizando BstF5I. Os isolados que mostraram esse padrão foram associados com os padrões IV e V. No entanto, não foi observada associação entre padrões polimórficos de RFLP e atividade de fosfolipase ou dados da população canina. Estes resultados demonstram a diversidade genética de M. pachydermatis e fornecem novas perspectivas sobre a epidemiologia destas espécies na população analisada.

ABSTRACT: Malassezia pachydermatis is a lipophilic and lipid-dependent yeast mostly isolated from animals’ skin; hence, it is regarded as a zoophilic species causing otitis externa in dogs. Aspects associated with its epidemiology and pathogenicity is a matter of interest. This study aimed to conduct a molecular characterization of 43 isolates of M. pachydermatis obtained from dogs with otitis externa. For this purpose, the 5.8S internal transcribed spacer 2 (ITS2) and D1/D2 26S rRNA regions were amplified, sequenced and analyzed using restriction fragment length polymorphism (RFLP) with AluI, CfoI, and BstF5I endonucleases. Phylogenetic analyses revealed that these isolates grouped with the sequence types I, IV and V, previously proposed for M. pachydermatis. Interestingly, we found a new polymorphic RFLP pattern using BstF5I, these isolates were associated with the sequence types IV and V, nevertheless an association between polymorphic RFLP patterns, and fosfolipase activity or canine population data was not observed. These findings underline the genetic diversity of M. pachydermatis and provide new insights about the epidemiology of this species in the analyzed population.

The present work was aimed at optimizing a culture medium for biomass production and phenolic compounds by using Ganoderma lucidum. The culture was optimized in two stages; a Plackett-Burman design was used in the first one for identifying key components in the medium and a central composite design was used in the second one for optimizing their concentration. Both responses (biomass and phenolic compounds) were simultaneously optimized by the latter methodology regarding desirability, and the optimal concentrations obtained were 50.00 g/L sucrose, 13.29 g/L yeast extract and 2.99 g/L olive oil. Maximum biomass production identified in these optimal conditions was 9.5 g/L and that for phenolic compounds was 0.0452 g/L, this being 100% better than that obtained in the media usually used in the laboratory. Similar patterns regarding chemical characterization and biological activity towards Aspergillus sp., from both fruiting body and mycelium-derived secondary metabolites and extracts obtained in the proposed medium were observed. It was shown that such statistical methodologies are useful for optimizing fermentation and, in the specific case of G. lucidum, optimizing processes for its production and its metabolites in submerged culture as an alternative to traditional culture.

Actinomicetos mineralizadores de fosfato involucrados en la interacción radical de Glomus sp. - trébol blanco. El objetivo de este trabajo fue determinar la capacidad de los actinomicetos para mineralizar fósforo orgánico y su interacción rizosférica con micorrizas en Trifolium repens L. (trébol blanco). En enero de 2009, en el laboratorio de la Unidad de Investigaciones Agropecuarias (UNIDIA) de la Pontificia Universidad Javeriana, Bogotá, Colombia, se cuantificó la actividad fosfatasa de Streptomyces sp. (cepa MCR9), Streptomyces sp. (cepa MCR26) y Thermobifida sp. (cepa MCR24) con la técnica del p-nitrofenil fosfato. Thermobifida sp. (cepa MCR24) presentó la menor actividad. La inoculación de solo actinomicetos presentó valores mayores de fósforo inorgánico en suelo, mientras que la concentración de fósforo a nivel foliar fue mayor cuando había co-inoculación actinomiceto-Glomus sp., a excepción de la cepa Thermobifida sp. (cepa MCR24). La co-inoculación con los actinomicetos y Glomus sp. estimuló la producción de biomasa y el porcentaje de colonización. Se determinó la capacidad para mineralizar fósforo orgánico por los actinomicetos al asociarse con Glomus sp. de forma sinérgica y benéfica tanto para la planta como para la micorriza. Se realizó el análisis de la varianza y prueba de “t” ALS (P<0,05) para comparar los promedios entre los tratamientos. Se determinó el grado de relación entre el porcentaje de micorrización y el peso seco de las plantas mediante “r” Coeficiente de Correlación, donde las plantas con los microorganismos presentaron una micorrización mayor del 70%.

Phosphate-mineralizing Actinomycetes involved in root interaction of Glomus sp.- white clover. The objective of this work was to determine the capacity of actinomicetes for organic phosphorus mineralization and their rhyzospere interaction with mycorrhiza of Trifolium repens L. (white clover). In January 2009, at the laboratory of the Unidad de Investigaciones Agropecuarias (UNIDIA) of the Pontificia Universidad Javeriana, Bogotá, Colombia, the phosphatase activity of Streptomyces sp. (strain MCR9), Streptomyces sp. (strain MCR26) and Thermobifida sp. (strain MCR24) were determined using the p-nitrofenil phosphate technique. Thermobifida sp. (strain MCR24) showed the lowest activity. Inoculation of actinomycetes alone resulted in higher values of inorganic soil P, while leaf P content was higher when co-inoculation of actinomycete-Glomus sp. occurred, with the exception of the strain Thermobifida sp. (strain MCR24). Co- inoculation with actynomicetes and Glomus sp. stimulated biomass production and colonization percent. Actynomicetes are sinergistic with Glomus sp., resulting in benefits for the plant and the mycorrhiza. Analysis of variance and “t” tests ALS (P<0,05) were conducted to compare the treatments. The relation between the degree of mycorrhization plant dry weight was determined by the “r” correlation coefficient, and plants associated with microorganisms was higher than 70%.

Objetivo. Determinar actividades amilolíticas, celulolíticas, lipolíticas, pectinolíticas y proteolíticas en 32 aislamientos de Fusarium spp. de origen humano, animal y vegetal. Materiales y métodos. Las actividades enzimáticas se determinaron a nivel cualitativo, por medio de la medición de halos de hidrólisis en placas de agar con el respectivo sustrato, y a nivel cuantitativo se realizó un cultivo líquido para determinar la degradación del respectivo sustrato por medio de técnicas colorimétricas. Resultados. Todos los aislamientos presentaron actividades enzimáticas a nivel cualitativo, excepto las amilolíticas y lipolíticas. La determinación a nivel cuantitativo fue posible para las enzimas evaluadas, a excepción de las lipasas. Conclusión. La determinación de los perfiles enzimáticos amilolíticos, celulolíticos, pectinolíticos y proteolíticos de cada uno de los aislamientos evaluados pertenecientes al género Fusarium sugirió su capacidad, indistintamente de su procedencia, de degradar estos sustratos.

Objective. To determine amylolytic, cellulolytic, lipolytic, pectinolytic and proteolytic activities in 32 Fusarium spp. isolates from humans, animals and plants. Materials and methods. Qualitative determination of enzymatic activities was done by measuring hydrolysis halos in agar plates with their corresponding substrate. Quantitative determination was done by colorimetric techniques, using liquid culture supernatants to determine the respective substrate degradation. Results. All isolates showed enzymatic activities from a qualitative point of view, except amylolytic and lipolytic. Quantitative determination was possible for all the evaluated enzymes except lipases. Conclusion. The determination of amylolytic, cellulolytic, pectinolytic and proteolytic enzymatic profiles of each of the Fusarium isolates assessed suggests their capacity to degrade these substrates, irrespectively of their origin.

Objetivo. Determinar as atividades amilolíticas, celulolíticas, lipolítica, pectinolíticas e proteolíticas em 32 isolamentos de Fusarium spp. de origem humana, animal e vegetal. Materiais e métodos. As atividades enzimáticas foram determinadas a nível qualitativo medindo os halos de hidrólise em placas de agar com o substrato respectivo e a nível quantitativo realizou-se uma cultura líquida para determinar a degradação do substrato respectivos por meio de técnicas colorimétricas. Resultados. Todos os isolados apresentaram uma atividade enzimática a nível qualitativo, exceto as amilolíticas e lipolíticas. A determinação a nível quantitativo foi possível para as enzimas testadas, exceto para as lipases. Conclusão. A determinação de perfis enzimáticos amilolíticos, celulolíticos, pectinolíticos e proteolíticos de cada um dos isolados testados pertencentes ao gênero Fusarium, sugeriu sua capacidade, independentemente da sua origem, para degradar estes substratos.