Óleo essencial de Pataqueira (Conobea scoparioides Cham. & Schltdl.) como conservante natural em Cosméticos / Pataqueira essential oil (Conobea scoparioides Cham. & Schltdl.) as a natural preservative in cosmetics

Um dos principais grupos de conservantes utilizados na maioria dos cosméticos são os parabenos que em muitos estudos demonstraram que podem provocar reações alérgicas como dermatite de contato, entre outras sensibilizações cutâneas. A fim de minimizar esses problemas, a indústria está produzindo cosméticos livres de conservantes ou de origem natural e em associações aos sintéticos. Dentre os conservantes naturais utilizados, podemos citar os óleos essenciais como uma alternativa viável. Diante deste contexto o presente trabalho visa avaliar experimentalmente o potencial antimicrobiano do óleo essencial de Conobea scoparioides Cham. & Schltdl., conhecida popularmente como pataqueira, o efeito de sua associação com parabenos e de sua eficácia como conservante em bases cosméticas. A composição do óleo essencial foi avaliada, indicando que este é composto em sua maior parte por terpenos, tendo éter metílico do timol (39,2%), timol (33,8 %) e α-felandreno (15,9%) como compostos majoritários. A atividade antimicrobiana do óleo essencial e do timol foi acessada através da concentração inibitória mínima (CIM), cujos resultados em µg/mL para o óleo essencial e o timol foram respectivamente: Staphylococcus aureus 650,70 e 284,90, Escherichia coli 721,53 e 271,20, Pseudomonas aeruginosa 1748,00 e > 2.000, Burkholderia cepacia 833,03 e 1.077,70, Candida albicans 521,43 e 172,61 e Aspergillus brasiliensis 300 e 400. O efeito sinérgico da associação do óleo essencial com os parabenos foi realizado através de um delineamento experimental centroide simplex para uma mistura de metilparabeno, propilparabeno e óleo essencial frente aos mesmos micro-organismos utilizados na determinação da atividade antimicrobiana. As concentrações ideais obtidas pela análise estatística para cada componente em µg/mL foram: 1120 para o metilparabeno, 350 para o propilparabeno e 675 para o óleo essencial. O teste de eficácia do sistema conservante em formulação cosmética foi efetuado empregando as concentrações ideais e mais duas concentrações superiores e uma abaixo do ideal. Para todas as cepas microbianas desafiadas o resultado do teste foi de redução total da carga microbiana inoculada nos sete dias de ensaio e nenhum aumento até o vigésimo oitavo dia o que demonstra a eficácia da associação do óleo essencial com os conservantes sintéticos. O óleo essencial de C. scoparioides apresentou um potencial antimicrobiano importante tanto sozinho como em associação com conservantes sintéticos. Estes resultados sugerem que esse óleo pode ser usado para compor um sistema conservante para formulações cosméticas contendo uma menor quantidade de sintéticos

One of the main groups of preservatives used in most cosmetics are parabens, that many studies have shown that they can cause allergic reactions such as contact dermatitis, among other skin sensitizations. To minimize these problems, the industry is producing cosmetics preservative free or using natural products instead and their combination with the synthetics. Among the natural preservatives used, we can mention essential oils as a viable alternative. In this context, the present work aims to experimentally evaluate the antimicrobial potential of the Conobea scoparioides Cham. & Schltdl. essential oil, popularly known as pataqueira, the effect of its association with parabens and its effectiveness as a preservative in cosmetic bases. The essential oil composition was analyzed, indicating that it is composed mostly of terpenes, with thymol methyl ether (39.2%), thymol (33.8%) and -phelandrene (15.9%) as major compounds. The antimicrobial activity of essential oil and thymol was accessed through the minimum inhibitory concentration (MIC), whose results in µg/mL for essential oil and thymol were respectively: Staphylococcus aureus 650.70 and 284.90, Escherichia coli 721, 53 and 271.20, Pseudomonas aeruginosa 1748.00 and > 2,000, Burkholderia cepacia 833.03 and 1,077.70, Candida albicans 521.43 and 172.61 and Aspergillus brasiliensis 300 and 400. The synergistic effect of the association of essential oil with parabens was performed through a centroid simplex experimental design for a mixture of methylparaben, propylparaben and essential oil against the same microorganisms used in the antimicrobial activity evaluation The ideal concentrations obtained by statistical analysis for each component in µg/mL were: 1120 for methylparaben, 350 for propylparaben and 675 for essential oil. The effectiveness test of the preservative system in cosmetic formulation was carried out using the ideal concentrations plus two higher concentrations and one below the ideal. For all challenged microbial strains, the test result was a total reduction of the inoculated microbial load in the seven days of testing and no increase until the twenty-eighth day, which demonstrates the effectiveness of the association of essential oil with synthetic preservatives. C. scoparioides essential oil showed an important antimicrobial potential both alone and in association with parabens. These results demonstrated that it can be used to compose a preservative system for cosmetic formulations containing lower amounts of synthetics

The occurrence of aflatoxigenic Aspergillus spp. in dairy cattle feed in Southern Brazil

ABSTRACT The presence of mycotoxins or related fungi in animal feed is a major problem for animal and human health. Silage and concentrated feed samples were collected from 21 dairy farms in the Western part of Paraná state in Southern Brazil. Water activity and pH of all samples were measured, and each sample was analyzed to check for the presence of aflatoxigenic Aspergillus. Water activity was observed to be lower in the concentrated feed samples. The pH was lower in the silage samples, indicating fermentation processes. Two silage samples and four concentrated feed samples were contaminated with Aspergillus spp. Seven isolates of Aspergillus spp. were obtained and their potential to produce aflatoxins was evaluated. Four of the isolates, two from the silage samples and two from the concentrated feed samples, produced the aflatoxins B1, B2, G1, and G2 in culture media. These isolates were identified as Aspergillus parasiticus and Aspergillus nomius. The presence of aflatoxigenic isolates of Aspergillus spp. in silage and concentrated feed samples is a matter of concern, because of the risk of aflatoxin production and contamination of the animal feed.

Aerobiología y biodeterioro del género Aspergillus Link en depósitos de tres instituciones patrimoniales cubanas / Aerobiology and biodeterioration of the genus Aspergillus Link in repositories of three Cuban heritage intitutions

Las especies del género Aspergillus Link constituyen importantes agentes biodeteriorantes de objetos de valor constructivo, cultural y patrimonial. En ambientes laborales, sus propágulos y metabolitos pueden provocar afecciones a la salud humana. Los estudios aeromicológicos pueden contribuir a minimizar el impacto de Aspergillus spp. en el biodeterioro y la calidad de vida. Se han caracterizado de acuerdo a su potencial biodeterió- geno y patogénico representantes de Aspergillus spp. en el aire de depósitos de materiales en tres instituciones patrimoniales cubanas. Las muestras se tomaron empleando un biocolector SAS súper 100. Se evaluó cualitativamente la actividad celulolítica, amilolítica y proteolítica de los aislados; así como la excreción de pigmentos y ácidos orgánicos. Se evaluó el crecimiento a 37°C y la secreción de hemolisinas. Aspergillus spp. constituyó parte considerable de la micobiota aérea viable en los depósitos con 22 especies correspondientes a 12 secciones, las de mayor variabilidad fueron Flavi y Restricti, mientras la primera y la sección Aspergillus fueron las mejor representadas. El 74 por ciento de las cepas mostraron alta capacidad para degradar los compuestos orgánicos evaluados y secretar ácidos, mientras el 34 por ciento excretó pigmentos de amarillo a pardo oscuro, se evidenció un amplio potencial biodeteriorante en general. Más del 30 por ciento de los aislados mostraron ser patógenos oportunistas peligrosos para el hombre al crecer a 37°C, secretar hemolisinas, y presentar un tamaño conidial que posibilita la depositación en alvéolos pulmonares. Las cepas con mayor impacto biodeteriorante y patogénico fueron las correspondientes a las Secciones Flavi, Nigri y la especie Aspergillus fumigatus Fresen.

Species of the genus Aspergillus Link cause damage to objects constructive, cultural and heritage value. In work environments, their propagules and metabolites may cause disorders to human health. Aerobiological studies can help minimize the impact of Aspergillus spp. in the biodeterioration and quality of life. Aspergillus spp. strains present in the air of the repositories in three Cuban heritage institutions were characterized according to their biodeteriogenic and pathogenic potential. Each isolated was classified taxonomically, cellulolytic, amylolytic and proteolytic activity as well as excretion of pigments and organic acids was evaluated. The capacity to grow at 37°C and secrete hemolysins was qualitatively evaluated. Aspergillus spp. was significant portion of viable air micobiota in the three repositories with 22 species corresponding to 12 sections, most variability were Flavi and Restricti, while the first section and Aspergillus were the best represented. The 74 percent of the strains showed high ability to degrade organic compounds evaluated and secrete acids, while 34 percent excreted pigments from yellow to dark brown, evidenced a broad biodeteriogenic potential in general. More than 30 percent of the isolates were shown to be dangerous opportunistic pathogens for humans to grow at 37°C, secrete hemolysin, and present a conidial size which enables the deposition pulmonary alveoli. Strains higher biodeteriogenic and pathogenic impact were those corresponding to the Flavi, Nigri sections and the species Aspergillus fumigatus Fresen.

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.

Susceptibility eest for fungi: clinical and laboratorial correlations in medical mycology / Teste de suscetibilidade para fungos: correlações clínico-laboratoriais em Micologia médica

SUMMARYDuring recent decades, antifungal susceptibility testing has become standardized and nowadays has the same role of the antibacterial susceptibility testing in microbiology laboratories. American and European standards have been developed, as well as equivalent commercial systems which are more appropriate for clinical laboratories. The detection of resistant strains by means of these systems has allowed the study and understanding of the molecular basis and the mechanisms of resistance of fungal species to antifungal agents. In addition, many studies on the correlation of in vitro results with the outcome of patients have been performed, reaching the conclusion that infections caused by resistant strains have worse outcome than those caused by susceptible fungal isolates. These studies have allowed the development of interpretative breakpoints for Candida spp. and Aspergillus spp., the most frequent agents of fungal infections in the world. In summary, antifungal susceptibility tests have become essential tools to guide the treatment of fungal diseases, to know the local and global disease epidemiology, and to identify resistance to antifungals.

RESUMONas últimas décadas, os testes de suscetibilidade a antifúngicos foram padronizados e, atualmente, servem tal como os testes de suscetibilidade a antibacterianos em laboratórios de microbiologia. Métodos de referência norte americanos e europeus foram desenvolvidos, assim como os equivalentes sistemas comerciais, estes últimos mais apropriados a laboratórios clínicos. A detecção de cepas resistentes por meio de tais sistemas permitiu o estudo e a compreensão das bases moleculares dos mecanismos de resistência de espécies fúngicas a fármacos antifúngicos. Além disso, foram realizados muitos estudos sobre a correlação de resultados obtidos in vitro com o desfecho clínico de pacientes permitindo a conclusão de que infecções por cepas resistentes têm pior evolução em relação às causadas por cepas sensíveis. Os estudos permitiram o estabelecimento de pontos de corte interpretativos (interpretative breakpoints development) para Candida spp. e Aspergillus spp., os agentes etiológicos mais frequentes de infecções fúngicas em todo o mundo. Em resumo, os testes de suscetibilidade representam uma ferramenta essencial para a orientação do tratamento de doenças fúngicas, para o conhecimento da epidemiologia local e global, bem como para a identificação de resistência a antifúngicos.

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.

Agentes comunes en las aspergilosis humanas: conceptos primarios en la diferenciación de sus complejos de especies / Common agent in human aspergillosis: primary concepts in their complex species differentiation

Las especies del género Aspergillus P.Micheli ex Haller, son consideradas como importantes organismos integrantes de la microbiota en distintos tipos de suelos en todas las latitudes. Presentan grandes capacidades fermentativas industriales, son agentes comunes en microbiología de alimentos, grandes biodescomponedores junto a las bacterias, contaminantes de productos agrícolas y de importancia en salud humana y animal. El género se asocia a 9 o más teleomorfos, formando un clado monofilético estrechamente relacionado con el género Penicillium. Debido al elevado número de especies, su dinámica taxonómica y sus frecuentes cambios en la nomenclatura, han complicado a menudo al investigador en algún campo de la biología, en especial cuando éste guarda relación con la industria o la medicina. Estos hongos productores de grandes cantidad de esporas secas de dispersión aérea, constituyen una fuente constante de exposición para los humanos, especialmente en los ambientes internos. Si bien es cierto, que los integrantes del género Aspergillus (en especial A. fumigatus) pueden causar un amplio espectro de enfermedades invasoras o no invasoras, la incidencia de otras especies del género reportadas en la literatura como agentes de aspergilosis, ha aumentado y complicado el trabajo taxonómico, genético y clínico en los últimos decenios. La presente revisión abarca el tema desde un punto de vista primario que incluye básicamente la morfofisiología de 34 especies involucradas en diversos casos clínicos geográficamente dispersos en todos los continentes. Se destacan algunos aspectos taxonómicos, ecológicos, patológicos, moleculares y el empleo de cultivos comunes en el diagnóstico en 3 de los subgéneros más frecuentes en clínica (Fumigati, Circumdati y Nidulantes) y 7 de sus secciones: Fumigati, Clavati, Nigri, Flavi, Terrei, Nidulantes y Usti...

Species of the genus Aspergillus P.Micheli ex Haller, are considered important members of the organisms of mycrobiota in different tipes of soils in all latitudes. They prsent large industrial fermentative capacity, are common agents in food microbiology, major biodeterioration agents with bacteria, contaminants in agricultural products, and important in human and animal health. The genus is associated with 9 or more teleomorphs, forming a monophyletic clade closely related to the genus Penicillium. Due to the high number of species, their taxonomic dynamic and frequent changes in nomenclature have often complicated the researcher in any field of biology, especially when related to industry or medicine. These fungi producing large amount of dry spores of aerial dispersion, and they are a constant source of exposure for humans, especially in indoor environments. While it is true, that members of the genus Aspergillus (especially A. fumigatus) can cause a wide spectrum invasive or non-invasive disease, the incidence of other species of the genus reported in the literature as agents of aspergillosis has increased and complicated the taxonomic, genetic and clinical work in recent decades. The present review covers the topic from a primary morphophysiology view and basically includes 34 species involved in various clinical cases geographically dispersed in all continents. It highlights some taxonomic, ecological, pathological and molecular aspects and the use of common culture media in the diagnosis in 3 of the most common clinical subgenera (Fumigati, Circumdati and Nidulantes) and 7 sections: Fumigati, Clavati, Nigri, Flavi, Terrei, Nidulantes and Usti...

Development of RFLP-PCR method for the identification of medically important Aspergillus species using single restriction enzyme MwoI

In this study we attempted to modify the PCR-RFLP method using restriction enzyme MwoI for the identification of medically important Aspergillus species. Our subjects included nine standard Aspergillus species and 205 Aspergillus isolates of approved hospital acquired infections and hospital indoor sources. First of all, Aspergillus isolates were identified in the level of species by using morphologic method. A twenty four hours culture was performed for each isolates to harvest Aspergillus mycelia and then genomic DNA was extracted using Phenol-Chloroform method. PCR-RFLP using single restriction enzyme MwoI was performed in ITS regions of rDNA gene. The electrophoresis data were analyzed and compared with those of morphologic identifications. Total of 205 Aspergillus isolates included 153 (75%) environmental and 52 (25%) clinical isolates. A. flavus was the most frequently isolate in our study (55%), followed by A. niger 65(31.7%), A. fumigatus 18(8.7%), A. nidulans and A. parasiticus 2(1% each). MwoI enabled us to discriminate eight medically important Aspergillus species including A. fumigatus, A. niger, A. flavus as the most common isolated species. PCR-RFLP method using the restriction enzyme MwoI is a rapid and reliable test for identification of at least the most medically important Aspergillus species.

Guía para el diagnóstico de laboratorio de enfermedad fúngica invasora por hongos filamentosos / Laboratory guidelines for diagnosis of invasive fungal disease caused by filamentous fungi

Early diagnosis and timely treatment are critical in Invasive Fungal Disease (IFD) caused by filamentous fungi in immunocompromised patients. Clinical features of IFD are nonspecific; therefore, images (CT scan), direct microscopic examination, staining and cultures of clinical samples and galactomannan determination in blood or BAL, are substantial. This guideline provides recommendations on transport and sample processing, including stains and culture media requirements. It also describes clues for diagnosis of major genera and species of Aspergillus.

La enfermedad fúngica invasora (EFI) por hongos filamentosos es causa de morbi-mortalidad en pacientes inmunocomprometidos, por lo que es fundamental el diagnostico precoz y su tratamiento oportuno. Las manifestaciones clínicas de las EFIs son inespecíficas, razón por la cual las imágenes (TAC), el examen microscópico directo, tinciones y cultivos de las muestras obtenidas y la determinación de galactomanano en sangre o en LBA tienen un rol fundamental. Esta guía tiene por objetivo recomendar las óptimas condiciones de transporte, procesamiento de las muestras, así como las tinciones y los medios de cultivos a utilizar. Se describen, además, claves para el diagnóstico de los principales géneros y especies de Aspergillus.

Variability in Galactomannan detection by platelia Aspergillus EIA™ according to the Aspergillus species / Variabilidade na detecção de galactomanana pelo Platelia Aspergillus EIA® de acordo com a espécie de Aspergillus

Here we investigate the extent to which different Aspergillus species release galactomannan (GM) in vitro. Marked variability was observed in GM reactivity between and within Aspergillus species, with A. terreus strains showing the highest GM indexes. The in vivo significance of these findings remains to be determined.

O estudo objetivou investigar a liberação in vitro de galactomanana (GM) em distintas espécies patogênicas de fungos do gênero Aspergillus. Grande variabilidade foi detectada tanto intra quanto inter espécies, sendo as cepas da espécie A. terreus relacionadas aos maiores índices de GM detectados. O significado in vivo destes achados permanece em aberto, porém merece investigação.

Ochratoxigenic fungi associated with green coffee beans (Coffea arabica L.) in conventional and organic cultivation in Brazil

The genera Aspergillus comprises species that produce mycotoxins such as aflatoxins, ochratoxins and patulin. These are cosmopolitan species, natural contaminants of agricultural products. In coffee grains, the most important Aspergillus species in terms of the risk of presenting mycotoxins belong to the genera Aspergillus Section Circumdati and Section Nigri. The purpose of this study was to assess the occurrence of isolated ochratoxigenic fungi of coffee grains from organic and conventional cultivation from the South of Minas Gerais, Brazil, as well as to evaluate which farming system presents higher contamination risk by ochratoxin A (OTA) produced by fungi. Thirty samples of coffee grains (Coffea arabica L.) were analysed, being 20 of them of conventional coffee grains and 10 of them organic. The microbiological analysis was done with the Direct Plating Technique in a Dichloran Rose Bengal Chloramphenicol Agar (DRBC) media. The identification was done based on the macro and micro morphological characteristics and on the toxigenic potential with the Plug Agar technique. From the 30 samples analysed, 480 filamentous fungi of the genera Aspergillus of the Circumdati and Nigri Sections were isolated. The ochratoxigenic species identified were: Aspergillus auricoumus, A. ochraceus, A. ostianus, A. niger and A. niger Aggregate. The most frequent species which produces ochratoxin A among the isolated ones was A. ochraceus, corresponding to 89.55%. There was no significant difference regarding the presence of ochratoxigenic A. ochreceus between the conventional and organic cultivation systems, which suggests that the contamination risk is similar for both cultivation systems.

Equinocandinas / Echinocandins

The echinocandins, caspofugin, micafungin, and anidulafungin, are lipopeptides that inhibit fungal growth by binding to β - (1.3) d glucan synthase. This enzyme is responsible for the formation of the peptidoglycan cell wall, and it is essential in fungi such as Candida spp, but less important in the case of Aspergillus and Fusarium species. We review the history, pharmacology and clinical trials that have showed clinical efficacy similar to amphotericin B for the management of fungal infections such as candidemia, invasive candidiasis and aspergillosis, even in cases refractory to initial treatment. These drugs have less toxicity and discontinuation is uncommonly required. Despite similar spectrum and tolerability, there are several pharmacological differences. Only a few clinical trials compare the clinical efficacy between them and their clinical application cannot be generalized. However, the echinocandins have demonstrated clinical efficacy in patients with invasive candidiasis and in others forms of systemic mycoses.

Las equinocandinas -caspofugina, micafungina y ani-dulafungina- son lipopéptidos que inhiben el crecimiento fúngico al unirse a la β-(1,3) d glucano sintetasa, enzima esencial para la síntesis en la pared celular de hongos como Candida spp, y menos importante en el caso de especies de Aspergillus y Fusarium. Se revisa la historia, farmacología y los diferentes ensayos clínicos que han evidenciado similar eficacia clínica a la de anfotericina B para el manejo de infecciones micóticas como candidemia, candidiasis invasora y aspergilosis, inclusive en casos refractarios al manejo inicial. Estos medicamentos tienen menor toxicidad y en pocos casos hay necesidad de retiro del tratamiento. Dado su espectro y tolerabilidad similar, su farmacología permite diferenciarlas. Se dispone de información limitada de estudios clínicos que las comparen entre ellas, limitando la extrapolación de la información a todo el grupo. Sin embargo, presentan eficacia clínica comprobada en pacientes con varias micosis invasoras.

Comparison of the activities of amphotericin B, itraconazole, and voriconazole against clinical and environmental isolates of Aspergillus species.

Background: Invasive fungal infections are a significant cause of morbidity and mortality in immunocompromised populations. Aims: To evaluate the susceptibility pattern of our isolates against amphotericin B, itraconazole, and voriconazole and to compare the antifungal activities of these agents with each other against the Aspergillus species tested. Settings and Design: A prospective study was designed to include clinical and environmental isolates of Aspergillus species. Materials and Methods: 420 sputum samples, 70 bronchoalveolar lavage fluids, 160 oral washings, and 47 environmental samples were collected. Direct microscopy by potassium hydroxide and lactophenol cotton blue mounts followed by culture on Sabourad`s dextrose agar (SDA) was done. Susceptibility testing was performed by the broth microdilution technique as per Clinical Laboratory Standards Institute standards (M-38A). Additionally, all the isolates were also tested by the colorimetric microdilution technique using Alamar Blue dye. Statistical Analysis: It was done by the Chi-square test and Z-test using SPSS statistical software version 12.0. Results and Conclusion: Twenty-seven isolates (47.3%) were recovered from patients with chronic bronchial asthma followed by fibrocavitary pulmonary tuberculosis in 9 (15.7%), allergic bronchopulmonary aspergillosis (ABPA) in 6 cases (10.5%), bronchiectasis in 3 (5.2%), bronchogenic carcinoma in 5 (8.7%) and those receiving radiotherapy for head and neck cancer 7 (12.2%). Thirteen environmental isolates were also included in the study. The most common isolate was A. fumigatus 28 (40%), followed by A. niger 22 (31%), A. flavus 13 (19%), and A. terreus 7(10%). All isolates were susceptible to amphotericin B, itraconazole, and voriconazole. Among the three agents tested, voriconazole exhibited lowest MICs (≤1 μg/ml) against all Aspergillus species.

Aportes morfotaxonómicos en el género Aspergillus link: claves para las especies ambientales y clínicas más comunes / Morphotaxonomic contributions in the genus Aspergillus Link: keys for the most common environmental and clinical species

Se condensan los principales alcances en el género Aspergillus y las metodologías de diagnóstico morfo-taxonómico de las especies más comunes de utilidad para el micólogo en áreas clínicas y ambientales. Se aporta información bibliográfica actualizada para las determinaciones polifásicas de las especies y algunas claves morfofisiológicas tentativas en ciertos taxa de importancia médica, con énfasis en la Sección Fumigati.

The main information about the Aspergillus genus together with methodologies for the morphotaxonomic diagnosis of the most common species useful for the mycologist in clinical and environmental areas are here condensed. An updated bibliography is forwarded in order to carry out polyphasic identification of the species as well as tentative morphophysiological keys in some taxa having medical significance with emphasis in Section Fumigati.

Characterization of Aspergillus species based on fatty acid profiles

Cellular fatty acid (FA) composition was utilized as a taxonomic tool to discriminate between different Aspergillus species. Several of the tested species had the same FA composition and different relative FA concentrations. The most important FAs were palmitic acid (C16:0), estearic acid (C18:0), oleic acid (C18:1) and linoleic acid (C18:2), which represented 95 percent of Aspergillus FAs. Multivariate data analysis demonstrated that FA analysis is a useful tool for differentiating species belonging to genus Aspergillus. All the species analyzed showed significantly FA acid profiles (p < 0.001). Furthermore, it will be possible to distinguish among Aspergillus spp. in the Flavi Section. FA composition can serve as a useful tool for the identification of filamentous fungi.

El género Aspergillus en la atmósfera de La Habana (Cuba) / The Aspergillus genus in the atmosphere of La Habana, Cuba

Se ha realizado un estudio de la contaminación fúngica ambiental en la atmósfera de la ciudad de La Habana (Cuba), durante el período de lluvias de los años 2001 y 2002, utilizando un método volumétrico viable, que ha permitido conocer la concentración de unidades formadoras de colonias (ufc/m3) totales y caracterizar los géneros y especies más abundantes, en especial del género Aspergillus. Durante el año 2001 las concentraciones más elevadas de hongos se obtuvieron en agosto y septiembre y las más bajas en julio, mientras que en el año 2002 destacan los niveles de septiembre y octubre frente a los escasos niveles del mes de agosto. Al analizar la densidad relativa de los géneros identificados se observa que Aspergillus es el género más abundante en la atmósfera seguido de Penicillium y Cladosporium. Las especies más abundantes en el año 2001 y relativamente constantes en el tiempo fueron A. japonicus, A. niger y A. flavus, mientras en el año 2002, los más abundantes fueron: A. japonicus, A. fumigatus y A. niger, siendo A. niger el único constante en el tiempo.

A study on environmental fungal contamination in the atmosphere of the City of La Habana (Cuba), during the rainy period in 2001 and 2002 was carried out. A viable volumetric method was used, making it possible to ascertain the concentration of units comprising total colonies (ufc/m3), and to characterize the most abundant genera and species, especially Aspergillus. Throughout 2001, the highest concentrations of fungi were collected in August and September, and the lowest in July; while in 2002, the highest levels were in September and October, compared to the minimal levels for the month of August. On analyzing the relative density of the genera identified, it was found that the most abundant genus in the atmosphere was Aspergillus, followed by Penicillium and Cladosporium. The most abundant species and relatively constant over time in 2001 were A. japonicus, A. niger and A. flavus while in 2002 A. japonicus, A. fumigates and A. niger; this specie was the only constant in time.

Reporte clínico: diagnóstico endoscópico e histológico de Aspergilosis Sinusal no invasiva en paciente inmunocompetente / Clinical report: an endoscopic and histologic diagnosis of a non invasive sinusal aspergillosis in an immunocompetent patient

Se describe un caso basado en un diagnóstico clínico, radiológico, endoscópico e histológico de una sinusitis crónica no invasiva en un paciente femenino de 40 años, con historia de rinorrea purulenta unilateral derecha de varios años de evolución. El TAC mostró velamiento del seno maxilar y calcificaciones con diagnóstico probable de infección micótica, sin embargo, no se solicitaron cultivos micológicos. Se indicó cirugía del seno maxilar mediante antrotomía media bajo endoscopia. El diagnóstico histológico reveló abundantes hifas septadas ramifi-cadas no invasivas de la mucosa sinusal con la presencia de algunas estructuras morfológicas asexuales que orientaron hacia un grupo taxonómico específico; Aspergillus sección Fumigati (posiblemente A. fumigatus). La evolución postoperatoria fue rápida y satisfactoria, sin requerir uso de antifúngicos, sólo aseos frecuentes de la cavidad operada hasta obtener una mucosa indemne y ventilada.

A case based on a clinical, radiologic, endoscopic and histologic diagnosis of a chronic non invasive sinusitis in a female 40 aged patient suffering from a purulent unilateral right rhinorrea which had been developing for several years is herein described. TAC revealed acertain masking of the maxilar sinus and calcifications which denoted a probable micotic infection yet mycological cultivations were not demanded. Surgery on the maxilar sinus by means of a medium antrotomy under endoscopy was indicated. The histologic diagnosisrevealed abundant non invasive septated, branched hyphae in the sinusal mucosa as well as some asexualmorphological structures which pointed to a particular taxonomic group; Aspergillus section Fumigati (possiblyA.fumigatus). Postoperatory evolution was fast and satisfactory since there was no need of using antifungaldrug, just frequent washing of the operated cavity until an undamaged and ventilated mucosa could be reached.

In-vitro susceptibility testing by agar dilution method to determine the minimum inhibitory concentrations of amphotericin B, fluconazole and ketoconazole against ocular fungal isolates.

PURPOSE: To standardize in-vitro antifungal susceptibility testing by agar dilution method to find out the minimum inhibitory concentration (MIC) of amphotericin B, fluconazole and ketoconazole on ocular fungal isolates. METHODS: A total of 180 ocular fungal isolates (130 filamentous fungi and 50 yeasts) were included. The antifungal drugs such as amphotericin B (0.0625-8 microg/mL), fluconazole (0.2-819.6 microg/mL) and ketoconazole (0.025-6.4 microg/mL) were incorporated in doubling dilutions in the yeast nitrogen base medium. The MIC was determined as the lowest concentration of the antifungal drug preventing growth of macroscopically visible colonies on drug containing plates when there was visible growth on the drug-free control plates. RESULTS: All 50 ocular isolates of yeast were susceptible to amphotericin B, while two (4%) and five (10%) strains were resistant to fluconazole and ketoconazole respectively. Of the 130 filamentous fungi tested, six (4.6%) were resistant to amphotericin B, 49 (37.7%) and 10 (7.6%) were resistant to fluconazole and ketoconazole respectively. Percentile 50 (MIC 50) and Percentile 90 (MIC 90) for all the three antifungal agents were calculated. Aspergillus niger, Aspergillus terreus and Candida krusei were found to be resistant to fluconazole and ketoconazole. CONCLUSION: This technique was found to be reliable, cost effective and easy to perform with consistent results.

Communities of P-solubilizing bacteria, fungi and arbuscular mycorrhizal fungi in grass pasture and secondary forest of Paraty, RJ - Brazil

Avaliaram-se as comunidades de bactérias, fungos solubilizadores de fosfato e fungos micorrízicos arbusculares em dois diferentes ecossistemas. Foram feitas amostragens em duas áreas de Mata Atlântica, em Paraty - RJ, Brasil, uma de floresta secundária e outra de pastagem. Foram avaliados quatro meios de cultura: GL (glicose e extrato de levedura), GES (glicose, extrato de solo, KNO3, CaCl2, MgSO4, NaCl, FeEDTA solução de micronutrientes), GAGES (glicose, extrato de solo, arabinose, glicerol, CaCl2, MgSO4 e NaCl) e GELP (glicose, extrato de solo, extrato de levedura, peptona, CaCl2, MgSO4 e NaCl) para isolamento de microrganismos solubilizadores de fosfato. A identificação das bactérias solubilizadoras foi baseada na análise do 16 S rDNA, enquanto que os fungos solubilizadores e os micorrízicos arbusculares foram identificados pela sua morfologia. O maior número de bactérias solubilizadoras foi obtido usando os meios GL e GELP. O maior número de fungos solubilizadores foi obtido com os meios GAGES e GES. As bactérias solubilizadoras foram identificadas como Enterobacteriaceae e Bacillus sp., e os fungos como Aspergillus sp. Glomus macrocarpum e Glomus etunicatum foram as espécies de fungos micorrízicos dominantes nas áreas de floresta secundária e pastagem, respectivamente.

Poblaciones de Aspergillus en semillas de maíz y soja de importación Argentina: énfasis en la sección Flavi / Aspergillus population in maize and soja seed imported from Argentina: ephasis on section Flavi

Durante un período de 5 meses se realizaron muestreos seriados en granos de maíz y soja de procedencia Argentina para estudiar la presencia de especies del género Aspergillus (en especial los de la sección Flavi). Para ambos tipos de granos se tomaron 10 muestras mensuales obtenidas de un puerto terrestre (Los Andes, Chile), desde submuestras transportadas en bolsas plásticas y proporcionadas por personal de control sanitario. Los granos se sembraron por duplicado en placas de Petri, utilizando 2 medios: agar malta sal (AMS) y agar agua sal (AAS). Todos los Aspergillus, fueron transferidos posteriormente en agar malta (AM) y agar czapek con extracto de levadura (CYA), para su determinación morfológica final, análisis poblacional, frecuencia de presencia en el tiempo y la posible producción de aflatoxinas (sección Flavi) en medio de cultivo específico. En maíz y soja y en ambos medios, se detectó un total de 1669 colonias del género Aspergillus (1193 en AMS y 476 en AAS), representando 18 taxa, (17 en maíz y 15 en soja), éstos fueron en órden decreciente: Aspergillus flavus, Eurotium amstelodami, E. rubrum, A.ellipticus, A.niger, A.candidus, A.ochraceus, A.terreus, A.versicolor, A.glaucus, A. tamarii, A.clavatus, A.ostianus, A. sydowii, A. fumigatus, A.parasiticus, A.wentii y A. sclerotiorum. Los 5 primeros taxa representaron el 85,7 por ciento de la presencia total. En general en maíz y soja la especie dominante fue A. flavus, mientras E. amstelodami y E. rubrum fueron más frecuentes en soja. Los valores más altos de presencia en maíz fueron en julio y en soja en noviembre. A.flavus presentó alta dominancia en ambos granos, seguida con baja frecuencia de A.tamarii y A. parasiticus. Para un análisis morfológico poblacional se seleccionaron al azar 50 cepas de A. flavus de todos los subtratos y muestras. En AM y CYA, se observaron cabezas radiadas a columnareslaxas mayoritariamente, aproximadamente la mitad de las cepas presentaron cabezas biseriadas y los...

Serial sampling of maize and soja grainsimported from Argentina were carried out for five months in order to study the occurrence of Aspergillus species (mainly those of Flavi section). For both kind of grains, ten monthly samples collected from a terrestrial port (Los Andes, Chile) were taken from subsamples transferred in plastic bags and supplied by sanitary control personnel. Grains were cultured induplicate on Petri dishes by using two media: malt salt agar (AMS)and salt water agar (AAS). All Aspergillusdetected were lately transferred in malt agar (AM) and czapek agar added with yeast extract (CYA) to determine its ultimate morphological character,population analysis, frequency of occurrence in time and possible production of aflotoxins in Flavi sectionin a particular medium of culture.In maize and soja and in both media,1669 colonies of the genus Aspergillus(1193 in AMS and 476 in AAS) was detected, representing 18 taxa, (17 in maize and 15 in soja), which in decreasing order were:Aspergillus flavus, Eurotium amstelodami, E.rubrum, A.ellipticus, A.niger, A.candidus, A.ochraceus, A.terreus, A.versicolor,A.glaucus, A.tamarii, A.clavatus, A.ostianus, A.sydowii, Afumigatus, A.parasiticus, A.wentii and A.sclerotium. The first five taxa represented 85.7 percent of the overall total. In general, the dominant species was A.flavus in maize and soja, whereas E.amstelodamiand E.rubrum were most frequent in soja. Highest values of occurrence in maize were in july and in november, in soja. A.flavus exhibited high dominancein both grains followed by a low frequency of A.tamarii and A.parasiticus. Fifty strains of A.flavus were randomly selected from every substrate and sample in order to carry out a population analysis. In AM and CYA, radiated heads to columnar laxa were mostly observed, about half strains exhibited biserial heads whereas 7-14day conidia (measured with an image processer) did not reveal any further change in...