Aspergillus andPenicillium (Eurotiales: Trichocomaceae) in soils of the Brazilian tropical dry forest: diversity in an area of environmental preservation

ResumenEl suelo es un sistema biológico complejo, que desempeña un papel fundamental en las plantas y los animales, especialmente en los bosques secos como la Caatinga. Los hongos del suelo, tales como Aspergillus y Penicillium, pueden ser utilizados como bioindicadores para la conservación de la biodiversidad. El objetivo de este estudio fue aislar e identificar las especies de Aspergillus y Penicillium del suelo, en los municipios de Ibimirim y Tupanatinga en el Parque Nacional Catimbau. Cinco colecciones se llevaron a cabo en cada área durante la estación seca de 2012, un total de 25 muestras de suelos por área. Los hongos fueron aislados mediante la suspensión en agua destilada estéril y se sembraron en medio de cultivo Agar Sabouraud más Cloranfenicol y Rosa de Bengala, y también en el medio Agar Dicloran Glicerol. Los aislamientos fueron identificados en el Laboratorio de Colección de Hongos y se confirmaron por secuenciación del espaciador transcrito interno de ADN. Un total de 42 especies fueron identificadas, 22 de ellas pertenecientes al género Aspergillus y 20 al género Penicillium. Los aislamientos de Penicillium mostraron una distribución uniforme en Tupanatinga con índices de uniformidad entre 0.92 y 0.88 en Ibimirim. Entre los aislamientos de Aspergillus el valor encontrado en Tupanatinga (0.85) fue muy similar al encontrado en Ibimirim (0.86). Se observó una gran diversidad y bajo predominio de hongos en las muestras de suelo. Estos resultados contribuyen a la estimación de la diversidad de hongos en ambientes secos, especialmente en la Caatinga, donde la diversidad es decreciente en los suelos que han sufrido alteraciones.

Abstract Soil is a complex biological system that plays a key role for plants and animals, especially in dry forests such as the Caatinga.Fungi from soils, such as Aspergillus and Penicillium, can be used as bioindicators for biodiversity conservation. The aim of this study was to isolate and identify species of Aspergillus and Penicillium in soil, from the municipalities of Tupanatinga and Ibimirim, with dry forests, in the Catimbau National Park. Five collections were performed in each area during the drought season of 2012, totaling 25 soil samples per area. Fungi were isolated by suspending soil samples in sterile distilled water and plating on Sabouraud Agar media plus Chloramphenicol and Rose Bengal, and Glycerol Dicloran Agar. Isolates were identified by morphological taxonomy in the Culture Collection Laboratory and confirmed by sequencing of the Internal Transcribed Spacer of rDNA. A total of 42 species were identified, of which 22 belong to the genus Aspergillus and 20 to Penicillium. Penicillium isolates showed uniform distribution from the collecting area in Tupanatinga, and the evenness indices found were 0.92 and 0.88 in Tupanatinga and Ibimirim, respectively. Among isolates of Aspergillus evenness, the value found in Tupanatinga (0.85) was very close to that found in Ibimirim (0.86). High diversity and low dominance of fungi in soil samples was observed. These results contributed to the estimation of fungal diversity in dry environments of the Caatinga, where diversity is decreasing in soils that have undergone disturbance. Rev. Biol. Trop. 64 (1): 45-53. Epub 2016 March 01.

Isolation and identification of toxigenic and non-toxigenic fungi in samples of medicinal plants from the market / Isolamento e identificação de fungos toxigênicos e não toxigênicos em amostras plantas medicinais do comércio

ABSTRACT: The consumption of preparations of medicinal plants has been increasing during the last decades in occidental societies. The presence of toxigenic fungi in a plant product may represent a potential risk of contamination, because of aflatoxins and ochratoxins. In this study, 12 samples of medicinal plants were analyzed in relation to the level of fungal contamination, and the presence of producers of ochratoxin A and aflatoxins was assessed by visualization of fungi using a cromatovisor in coconut milk. Most of the species found belong to the genus Cladosporium, Fusarium, Aspergillus and Penicillium. Species producing ochratoxin A were present in 2 samples (16.7%), Melissa and Hibiscus. Species producing aflatoxin were found in samples of Jacaranda decurrens (8.33%). This study suggests that herbs, if stored improperly, can provide the growth of fungi and should be examined before consumption.

RESUMO: O consumo das plantas medicinais vem aumentando nas últimas décadas nas sociedades ocidentais, porém, a presença de fungos toxigênicos nestas plantas pode representar um risco em potencial de contaminação devido à produção de aflatoxinas e ocratoxinas. Neste trabalho, 12 amostras de plantas medicinais foram analisadas em relação ao nível de contaminação por fungos, enquanto a presença de produtores de ocratoxina A e aflatoxinas foi avaliada pela visualização em cromatovisor dos fungos em meio de leite de coco. A maioria das espécies encontradas pertence aos gêneros Cladosporium, Fusarium, Aspergillus e Penicillium. Espécies produtoras de ocratoxina A estavam presentes em 2 amostras (16,7%), Melissa e Hibisco. Espécies produtoras de aflatoxina foram encontradas na amostra de Carobinha (8,33%). Este trabalho sugere que as ervas, sendo armazenadas inadequadamente, proporcionam o crescimento de fungos e, por isso, estes devem ser examinados antes do consumo.

Characterization and antimicrobial activity of lectins from Penicillium sp.

Ten Penicillium sp. were screened for lectin activity for occurrence of lectins. Mycelial extracts from submerged cultures of P. corylophilum, P. expansum and P. purpurogenum showed agglutination against human (A, B, AB and O), goat, sheep, pig and rabbit erythrocytes. Neuraminidase treatment to human blood type O erythrocytes substantially increased their agglutinability by all the lectins as compared to untreated erythrocytes. Modification of erythrocyte surfaces by protease increased the lectin titre only of P. corylophilum with no effect on other two lectins. P. corylophilum and P. expansum displayed relatively lower titres in mycelial extracts prepared from agar plate cultures as compared to broth cultures. A panel of sugars was tested for inhibition of lectin activity. All the lectins were found to be specific for asialofetuin, bovine submaxillary mucin, porcine stomach mucin, chondroitin-6-sulphate, D-sucrose and D-glucose. P. corylophilum lectin was expressed (Titre 8) by 5 day old cultures, reaching its maximum level (Titre 32) upon 8 days of cultivation, thereafter declin in lectin activity was observed. P. purpurogenum lectin was expressed by 7-10 days old cultures, while in P. expansum maximum lectin activity was elaborated by 5-8 days old cultures. Lectin extracts from all the three species were found to possess antimicrobial activities. Lectin extracts from the three Penicillium species displayed antifungal activity and antibacterial activity against Gram-negative and Gram-positive bacterial strains.

Penicillium species present in Uruguayan salami / Especies de Penicillium presentes en salamines uruguayos

The surface coverage of certain dry fermented sausages such as Italian salami by some species of Penicillium provides their characteristic flavor and other beneficial properties. One of them is the protective effect by means of a uniform film of white mold against undesirable microorganisms. The aim of this work was to identify and to isolate the fungal species present in mature Italian type of salami and to evaluate if it is possible to obtain some of them as starters. In addition, the effects of temperature (14 °C and 25 °C), water activity (a w) (0.90, 0.95 and 0.995) and 2.5 % sodium chloride (NaCl) on fungal growth were determined. Similarly, the proteolytic and lipolytic activity and the ability to produce toxic secondary metabolites were evaluated in order to characterize some possible starter strain. All species found belong to the genus Penicillium, including a performing starter as Penicillium nalgiovense and some potentially toxicogenic species. All the strains showed a higher growth rate at 25 °C. The production of extracellular proteases and lipases was significantly higher at 25 °C than at 14 °C with and without sodium chloride. Only Penicillium expansum produced patulin. On the other hand, Penicillium griseofulvum was the only species that produced ciclopiazonic acid but none of the strains produced penicillin. The species present on salami, Penicillium nalgiovense, Penicillium minioluteum, Penicillium brevicompactum and Penicillium puberulum were unable to produce any of the evaluated toxins. These findings suggest that some fungal isolates from the surface of salami such as P. nalgiovense are potentially useful as starters in sausage manufacture.

La cobertura de la superficie de los embutidos fermentados secos -como el salamín tipo italiano- por algunas especies de Penicillium les proporciona un sabor característico y otras propiedades beneficiosas. Una de ellas es el efecto de protección contra microorganismos indeseables, al formarse una película blanca uniforme de mohos. El objetivo de este trabajo fue aislar e identificar los hongos filamentosos encontrados en la superficie de salamines tipo italiano y evaluar la posibilidad de obtener especies para utilizarse como cultivos iniciadores. Se determinó el efecto de la temperatura, la actividad de agua y del cloruro de sodio sobre el crecimiento fúngico. La actividad proteolítica y lipolítica y la capacidad de producir metabolitos secundarios tóxicos fueron evaluadas con el fin de caracterizar algunos posibles cultivos iniciadores. Todas las cepas fúngicas aisladas e identificadas correspondieron a especies del género Penicillium, algunas benéficas, como Penicillium nalgiovense, y otras potencialmente toxicogénicas. Estas cepas tuvieron diferentes tasas de crecimiento en respuesta a las diferentes condiciones de cultivo. Todas las cepas mostraron mayor crecimiento a 25 °C. La producción de proteasas y lipasas extracelulares fue significativamente mayor a 25 °C que a 14 °C. Penicillium expansum fue la única especie que produjo patulina y Penicillium griseofulvum fue la única que produjo ácido ciclopiazónico. Ninguna de las especies produjo penicilina. Penicillium nalgiovense, Penicillium minioluteum, Penicillium brevicompactum y Penicillium puberulum no produjeron ninguna de las toxinas evaluadas. Estos resultados sugieren que algunos aislamientos fúngicos, como P. nalgiovense, son potencialmente útiles como cultivos iniciadores en la fabricación de estos productos.

Infección diseminada por Penicillium marneffei en un paciente HIV-positivo: Primera observación en la República Argentina / Disseminated infection due to Penicillium marneffei related to HIV infection: First observation in Argentina

Se presenta el primer caso humano de peniciliosis por Penicillium marneffei observado en la República Argentina. El paciente era un joven de 16 años, HIV-positivo, procedente de un área rural del sur de China. El paciente fue internado en el Hospital "F. J. Muñiz" por padecer una neumonía grave con insufciencia respiratoria aguda. El agente causal fue aislado de un lavado broncoalveolar y se lo observó en un citodiagnóstico de piel. La identifcación de P. marneffei fue confrmada por las características fenotípicas del aislamiento y la amplifcación del ADNr. El enfermo padecía una infección muy avanzada por HIV que condujo a la aparición simultánea de infecciones por citomegalovirus, Pneumocystis jirovecii y procesos bacterianos nosocomiales. Este complejo cuadro derivó en una evolución fatal.

The frst case observed in Argentina of AIDS-related human penicillosis is herein presented. The patient was a six- teen year-old young man coming from a rural area of southern China. He was admitted at the F. J. Muñiz Hospital of Buenos Aires city with severe pneumonia and adult respiratory distress. Penicillium marneffei was isolated from bronchoalveolar lavage fuid and was microscopically observed in a skin cytodiagnosis. P. marneffei identifcation was confrmed by rRNA amplifcation and its phenotypic characteristics. The patient suffered an advanced HIV infection and also presented several AIDS-related diseases due to CMV, nosocomial bacterial infections and Pneumocystis jirovecii which led to a fatal outcome.

Novel antimicrobial secondary metabolites from a Penicillium sp. isolated from Brazilian cerrado soil / Nuevos metabolitos antimicrobianos secundarios de Penicillium sp. aislada en suelo del cerrado de Brasil

The morphological features of a Penicillium, isolated from Brazilian cerrado soil, were characterized and showed to be distinctly different from all well-defined Penicillium species. Chemical and biological investigation on the ethyl acetate extract of this Penicillium isolate resulted in the isolation of three new naphthalenoids: a major metabolite, methyl 6-acetyl-4-methoxy-5,7,8-trihydroxynaphthalene-2-carboxylate and two minor ones, methyl 6-acetyl-4-methoxy-7,8-dihydroxynaphthalene-2-carboxylate and methyl 6-acetyl-4-methoxy-5,8-dihydroxynaphthalene-2-carboxylate. Their structures were determined based on their mono and bidimensional nuclear magnetic resonance data. Acetyl, allyl and methoxyl derivatives of the major metabolite were prepared in order to establish structure-activity relation. Antimicrobial activity of the major natural product and its semi-synthetic derivatives was screened by macro dilution methodology and the corresponding minimum inhibitory concentrations were determined. Natural secondary metabolite methyl 6-acetyl-4-methoxy-5,7,8-trihydroxynaphthalene-2-carboxylate, isolated in a very high yield (0.3175 mg.L-1) showed to be the most active compound, possessing expressive activity against Candida albicans (minimum inhibitory concentration (MIC) 32 ug/mL), Listeria monocitogenes and Bacillus cereus (MIC 64 µg/mL for both).

Penicillium marneffei--an indicator disease of AIDS: a case report.

Penicillium marneffei (PM), the only dimorphic species of the genus penicillium is the etiological agent of penicilliosis marneffei. This opportunistic fungal infection occurs among human immunodeficiency virus (HIV) infected and other immunocompromised patient in several regions of South-east Asia, where the infection is considered as an indicator disease of AIDS. A case of penicilliosis marneffei is reported in a patient whose HIV status was unknown and later turned to be in the late stage of AIDS. This demonstrates that it is indeed an indicator disease of AIDS. In India, penicilliosis has been reported among the inhabitants of Manipur state where the prevalence of HIV infection / AIDS is very high. The causative agent was first isolated from a captive bamboo rat. Investigation of the prevalence of the organism among bamboo rats of different countries of South East Asia revealed four species of bamboo rats to be harboring the organism. These four species of bamboo rats are Rhizomys sinensis, R. pruinosus, R. sumatrensis and Cannomys badius. In Manipur, Penicillium marneffei has been isolated from Cannomys badius. Any patient presenting with penicilliosis marneffei should be subjected to HIV counselling and testing if the HIV status is not known and further study regarding the ecology and epidemiology of the fungus is needed.

Actividades hidrolíticas y caracterización isoenzimática de poblaciones microbianas aisladas del patrimonio documental del Archivo General de Colombia / Hydrolytic activity and isoenzymatic characterization of microbial population Isolated of documental Heritage of General Archive of Colombia (AGN)

Este trabajo presenta un modelo para el aislamiento y evaluación de microorganismos agentes deteriorantes del acervo documental. Se recuperaron e identificaron 28 aislamientos tanto de las unidades de conservación con biodeterioro como de la atmósfera del depósito 15 del Archivo General de la Nación de Colombia (AGN). De este grupo, 16 aislamientos microbianos mostraron capacidad hidrolítica sobre fibras vegetales cuando se cultivaron en medios con paredes celulares al 1porciento como única fuente de carbono. A las poblaciones microbianas recuperadas se les evaluó su capacidad para hidrolizar celulosa, xilano, almidón y proteínas, considerando los halos de degradación y el número de sustratos hidrolizados, se seleccionaron cuatro aislamientos: dos de Penicillium sp., uno de Bacillus sp. y uno de Actinopolyspora sp. A los aislamientos se les cuantificó las actividades depolimerasas y accesorias y se les determinó el perfil isoenzimático para celulasas y xilanasas. Los resultados sugieren: (i) los hongos filamentosos y actinomycetes son más eficientes en la degradación de polímeros complejos, (ii) posiblemente las poblaciones bacterianas actúan como colonizadores secundarios y (iii) el perfil isoenzimático permite descartar microorganismos saprófitos, de especializados en degradar soporte celulolítico

Morphological variation in pathogenic strains of Penicillium marneffei.

Penicillium marneffei is a dimorphic fungus known to be pathogenic to animals and man. The natural reservoir of this organism was known to be bamboo rats found in South Vietnam, Thailand and China. The first two human infections were reported in 1959 and 1973 from the United States. Up to 1984, five new cases of human penicillosis were reported from Thailand. Since then several more cases have been reported from different parts of the world mainly from the southern part of China. However, there are very limited mycological descriptions of this fungi. In this report, five Thai strains were studied for colonial morphology in comparison with Reference strain PLM 689. Variation in mycelial pigment was observed ranging from yellowish-green to orange with water soluble red pigment produced in every strain which can be seen early from the reverse side. Ultrastructural study by both scanning electron microscopy (SEM) and transmission electron microscopy (TEM) was compared with that of the reference strain PLM 689. PLM 689 strain had only biverticillate penicilli, but all five strains from Thailand had both monoverticillate and biverticillate penicilli which occasionally appeared on the same branch. The conidia of the Thai isolates were oval in shape and 1.3-2 x 0.7-1.6 microns in size smaller than those of PLM 689 which were 2.5-4 x 2-3 microns. Phialides were also smaller and a little shorter but the number of phialides was similar to those of PLM 689 ranging 4-10 except for one strain which had 3-16 phialides. All Thai strains have stipes smaller and somewhat longer than those of PLM 689.(ABSTRACT TRUNCATED AT 250 WORDS)