Fungal decomposition of chicken-feather waste in submerged and solid-state fermentation / Decomposição fúngica de resíduos de pena de frango em fermentação submersa e de estado sólido

Poultry industry is expanding rapidly and producing million tons of feather waste annually. Massive production of keratinaceous byproducts in the form of industrial wastes throughout the world necessitates its justified utilization. Chemical treatment of keratin waste is proclaimed as an eco-destructive approach by various researchers since it generates secondary pollutants. Keratinase released by a variety of microbes (bacteria and fungi) can be used for the effective treatment of keratin waste. Microbial degradation of keratin waste is an emerging and eco-friendly approach and offers dual benefits, i.e., treatment of recalcitrant pollutant (keratin) and procurement of a commercially important enzyme (keratinase). This study involves the isolation, characterization, and potential utility of fungal species for the degradation of chicken-feather waste through submerged and solid-state fermentation. The isolated fungus was identified and characterized as Aspergillus (A.) flavus. In a trial of 30 days, it was appeared that 74 and 8% feather weight was reduced through sub-merged and solid-state fermentation, respectively by A. flavus. The pH of the growth media in submerged fermentation was changed from 4.8 to 8.35. The exploited application of keratinolytic microbes is, therefore, recommended for the treatment of keratinaceous wastes to achieve dual benefits of remediation.

A indústria avícola está se expandindo rapidamente e produzindo milhões de toneladas de resíduos de penas anualmente. A produção massiva de subprodutos queratinosos na forma de resíduos agrícolas e industriais em todo o mundo exige sua utilização justificada. O tratamento químico de resíduos de queratina é proclamado como uma abordagem ecodestrutiva por vários pesquisadores, uma vez que gera poluentes secundários. A queratinase liberada por uma variedade de micróbios (bactérias e fungos) pode ser usada para o tratamento eficaz de resíduos de queratina. A degradação microbiana de resíduos de queratina é uma abordagem emergente e ecológica e oferece benefícios duplos, ou seja, tratamento de poluente recalcitrante (queratina) e obtenção de uma enzima comercialmente importante (queratinase). Este estudo envolve o isolamento, caracterização e utilidade potencial de espécies de fungos para a degradação de resíduos de penas de frango por meio da fermentação submersa e em estado sólido. O fungo isolado foi identificado e caracterizado como Aspergillus (A.) flavus. Em um ensaio de 30 dias, constatou-se que 74% e 8% do peso das penas foram reduzidos por A. flavus, respectivamente, por meio da fermentação submersa e em estado sólido. O pH do meio de crescimento em fermentação submersa foi alterado de 4,8 para 8,35. A aplicação explorada de micróbios queratinolíticos é, portanto, recomendada para o tratamento de resíduos ceratinosos para obter benefícios duplos de remediação.

Desarrollo y capacidad de remoción de los pesticidas atrazina, clorpirifós y endosulfán por parte de cepas de Aspergillus sección Flavi no toxigénicas / Development and removal ability of non-toxigenic Aspergillus section Flavi in presence of atrazine, chlorpyrifos and endosulfan

This study evaluated the in vitro effect of three concentrations of atrazine, chlorpyrifos and endosulfan on the growth parameters of four non-toxigenic Aspergillus section Flavi strains. The ability of the strains to remove these pesticides in a synthetic medium was also determined. Growth parameters were measured on soil extract solid medium supplied with 5,10 and 20mg/l of each pesticide, and conditioned to -0.70, -2.78, -7.06 and -10.0 water potential (MPa). Removal assays were performed in Czapek Doc medium (CZD) supplied with 20mg/l of each pesticide under optimal environmental conditions (-2.78 of MPa and 25 °C). The residual levels of each pesticide were detected by the reversed-phase HPLC/fluorescence detection system. The lag phases of the strains significantly decreased in the presence of the pesticides with respect to the control media. This result indicates a fast adaptation to the conditions assayed. Similarly, the mycelial growth rates in the different treatments increased depending on pesticide concentrations. Aspergillus oryzae AM 1 and AM 2 strains showed high percentages of atrazine degradation (above 90%), followed by endosulfan (56 and 76%) and chlorpyrifos (50 and 73%) after 30 days of incubation. A significant (p <0.001) correlation (r = 0.974) between removal percentages and growth rate was found. This study shows that non-toxigenic Aspergillus section Flavi strains from agricultural soils are able to effectively grow in the presence of high concentrations of atrazine, chlorpyrifos and endosulfan under a wide range of MPa conditions. Moreover, these strains have the ability to remove high levels of these pesticides in vitro in a short time.

En este estudio se evaluó los efectos in vitro de 3 concentraciones de atrazina, clorpirifós y endosulfán sobre los parámetros de crecimiento de 4 cepas no toxigénicas de Aspergillus sección Flavi. También se evaluó la capacidad de las cepas de remover los pesticidas. Los parámetros de crecimiento se ensayaron en medio agar extracto de suelo suplementado con 5, 10 y 20mg/l de cada pesticida y acondicionado a -0.70, -2.78, -7.06 y -10.0 de potencial de agua (MPa). Los ensayos de remoción se realizaron en medio Czapek Dox con 20mg/l de cada pesticida bajo condiciones óptimas de crecimiento (-2.78 de MPa y 25 °C). Los niveles residuales de atrazina, clorpirifós y endosulfán se detectaron en un sistema HPLC con detección por fluorescencia. La fase de latencia de las cepas disminuyó significantemente en presencia de los pesticidas, indicando una rápida adaptación a dichas condiciones. La velocidad de crecimiento se incrementó considerablemente dependiendo de la concentración de pesticida. Las cepas Aspergillus oryzae AM1 y AM2 mostraron porcentajes elevados de degradación de atrazina (aproximadamente el 90%), seguidos por endosulfán (56 y 76%) y clorpirifós (50 y 73%). Se observó una correlación (r = 0.974) significante (p <0.001) entre el porcentaje de pesticida removido y la velocidad de crecimiento. Este estudio muestra que cepas no-toxigénicas de Aspergillus sección Flavi aisladas de suelos agrícolas desarrollan eficientemente en presencia de altas concentraciones de atrazina, clorpirifós y endosulfán en un amplio rango de MPa. Además, presentan capacidad de remover in vitro altos niveles de pesticidas en corto tiempo.

Differential role of gpaB and sidA gene expressions in relation to virulence in Aspergillus species from patients with invasive aspergillosis

Abstract The virulence genes in invasive aspergillosis (IA) have not been analyzed adequately. The present study was designed to evaluate the expression of gpaB and sidA genes, which are important virulence genes in Aspergillus spp. from bronchoalveolar lavage (BAL) samples. Direct examination and culture on Czapek Agar and Sabouraud Dextrose Agar media were performed for 600 BAL specimens isolated from patients with possible aspergillosis. A Galactomannan ELISA assay was also carried out. The expression levels of the gpaB and sidA genes in isolates were analyzed using quantitative real-time PCR (qRT-PCR). We identified 2 species, including Aspergillus flavus (A. flavus) and Aspergillus fumigatus (A. fumigatus) in 25 positive samples for invasive aspergillosis as validated using GM-ELISA. A. flavus is the main pathogen threatening transplant recipients and cancer patients worldwide. In this study, A. flavus had low levels of the gpaB gene expression compared to A. fumigatus (p = 0.006). The highest sidA expression was detected in transplant recipients (p = 0.05). There was no significant correlation between sidA expression and underlying disease (p = 0.15). The sidA and gpaB gene expression patterns may provide evidence that these virulence genes play important roles in the pathogenicity of Aspergillus isolates; however, there are several regulatory genes responsible for the unexpressed sidA and gpaB genes in the isolates.

In vitro antifungal susceptibility of clinical and environmental isolates of Aspergillus fumigatus and Aspergillus flavus in Brazil

ABSTRACT The in vitro susceptibility of 105 clinical and environmental strains of Aspergillus fumigatus and Aspergillus flavus to antifungal drugs, such as amphotericin B, azoles, and echinocandins was evaluated by the broth microdilution method proposed by the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Following the EUCAST-proposed breakpoints, 20% and 25% of the clinical and environmental isolates of A. fumigatus, respectively, were found to be resistant to itraconazole (Minimal Inhibitory Concentration, MIC > 2.0 mg/L). Voriconazole showed good activity against A. fumigatus and A. flavus strains, except for one clinical strain of A. fumigatus whose MIC was 4.0 mg/L. Posaconazole (≤0.25 mg/L) also showed appreciable activity against both species of Aspergillus, except for six A. fumigatus strains with relatively higher MICs (0.5 mg/L). The MICs for Amphotericin B ranged from 0.06 to 1.0 mg/L for A. fumigatus, but were much higher (0.5-8.0 mg/L) for A. flavus. Among the echinocandins, caspofungin showed a geometric mean of 0.078 and 0.113 against the clinical and environmental strains of A. flavus, respectively, but had elevated minimal effective concentrations (MECs) for seven of the A. fumigatus strains. Anidulafungin and micafungin exhibited considerable activity against both A. fumigatus and A. flavus isolates, except for one environmental isolate of A. fumigatus that showed an MEC of 1 mg/L to micafungin. Our study proposes that a detailed investigation of the antifungal susceptibility of the genus Aspergillus from different regions of Brazil is necessary for establishing a response profile against the different classes of antifungal agents used in the treatment of aspergillosis.

Hongos productores de micotoxinas asociados a hierbas medicinales en la Ciudad de São Paulo, Brasil / Mould and mycotoxins associated in medicinal plants in São Paulo, Brazil

Antecedentes: la fitoterapia es una de las más antiguas prácticas utilizadas por la humanidad. Hasta mediados del siglo XIX, cuando se introdujeron los medicamentos, la formulación de estos generalmente era basada en plantas medicinales. Objetivos: Determinar la micobiota y los niveles de aflatoxinas originadas de Aspergillus sección Flavi aislados de las 50 muestras de medicamentos fitoterápicos comercializados actualmente en la ciudad de São Paulo, Brasil. Métodos: Cincuenta (50) muestras de medicamentos fitoterápicos en la forma de hojas (té-25) y cápsulas (25) fueron colectadas de agosto de 2000 a julio de 2001. Los hongos filamentosos aislados fueron identificados al nivel de género de acuerdo con las características morfológicas y criterios taxonómicos. El análisis de aflatoxinas fue realizada por cromatografía de capa fina (TLC). Resultados: El análisis microbiológico mostró que 41 (82 por ciento) de los medicamentos fitoterápicos presentaron un crecimiento fúngico sobre las 100 UFC/g. Un total de 106 especies de seis diferentes géneros fueron aislados (Aspergillus, Penicillium, Mucor, Rhizopus y Alternaria). El género Aspergillus fue el predominante (60.5 por ciento) seguido por Penicillium (20,0 por ciento). Aspergillus niger (30 por ciento) A. flavus (22 por ciento), A. fumigatus (6,5 por ciento) y A. parasiticus fueron las especies de Aspergillus identificadas. Se observó que 13 (56,5 por ciento), de los 23 A. flavus aislados y dos aislados de A. parasiticus produjeron aflatoxinas. Conclusiones: La contaminación observada en la mayoría de los productos y el alto nivel de cepas productoras de aflatoxinas justifica un análisis más cuidadoso de los medicamentos fitoterápicos comercializados y la aplicación de leyes más rigurosas son necesarias para garantizar la calidad de los productos.

Background: phytotherapy is one of the most ancient practices used by humanity. In Antiquity until the middle of the XIX century, when chemotherapeutic drugs were introduced, formulation of medicines was usually based on medicinal plants. Objective: To determine mycobiota and levels of Aspergillus section Flavi aflatoxins isolated from 50 samples of phytotherapeutic remedies currently commercialized in São Paulo, Brazil. Methods: Fifty (50) samples of phytotherapeutic remedies in the form of leaves (teas-25) and powders (capsules-25) were collected from August 2000 to July 2001. Filamentous fungi isolates were identified at the genera level in accordance with morphological characteristics and taxonomic criteria. Aflatoxins were performed by Thin-layer chromatography (TLC). Results: The microbiological analysis showed that 41 (82 percent) of phytotherapeutic remedies presented a fungal growth over 100 CFU/g. A total of 106 species of six different genera were isolated (Aspergillus, Penicillium, Mucor, Rhizopus and Alternaria). The genus Aspergillus was the predominant (60.5 percent) followed by Penicillium genus (20.0 percent). Aspergillus niger (30 percent) A. flavus (22 percent), A. fumigatus (6.5 percent) and A. parasiticus were the species of Aspergillus identified. It was observed that 13 (56.5 percent) of 23 A. flavus isolates and two A. parasiticus isolates produced aflatoxins. Conclusions: The contamination observed in most products and the high level of aflatoxigenic strains justify the concern regarding the execution of more careful analyzes of the commercialized phytotherapeutic remedies and the application of more rigorous laws that may warrant the quality of these products.

The use of chitosan in protecting wooden artifacts from damage by mold fungi

Background: Many buildings in Egypt e.g. museums, mosques and churches, do not possess controlled environments for minimizing the risks of damage of wooden artifacts due to the growth of fungi. Fungal damage usually appears as change in wood color, appearance of stains, and sometimes deformation of wooden surfaces. In this study we focused on the effect that some fungi exert on the properties of wooden artifacts and evaluated the effectiveness of different concentrations of chitosan on their protection against damage by mold fungi. Results: Samples were collected from different monuments and environments, and fungi growing on them were isolated and identified. The isolated Penicillium chrysogenum, Aspergillus flavus and /Aspergillus niger strains were used for the infestation of new pitch pine samples. The results revealed that the lightness of samples infected with any of the tested fungi decreased with increasing incubation times. XRD analysis showed that the crystallinity of incubated samples treated individually with the different concentrations of chitosan was lower than the crystallinity of infected samples. The crystallinity index measured by the first and the second method decreased after the first and second months but increased after the third and fourth months. This may due to the reducing of amorphous part by enzymes or acids produced by fungi in wooden samples. Conclusions: The growth of fungi on the treated wood samples decreased with increasing the concentration of chitosan. Hence, it was demonstrated that chitosan prevented fungal growth, and its use could be recommended for the protection of archeological wooden artifacts.

Molecular characterisation of Aspergillus flavus isolates from peanut fields in India using AFLP

Aflatoxin contamination of peanut, due to infection by Aspergillus flavus, is a major problem of rain-fed agriculture in India. In the present study, molecular characterisation of 187 Aspergillus flavus isolates, which were sampled from the peanut fields of Gujarat state in India, was performed using AFLP markers. On a pooled cluster analysis, the markers could successfully discriminate among the ‘A’, ‘B’ and ‘G’ group A. flavus isolates. PCoA analysis also showed equivalent results to the cluster analysis. Most of the isolates from one district could be clustered together, which indicated genetic similarity among the isolates. Further, a lot of genetic variability was observed within a district and within a group. The results of AMOVA test revealed that the variance within a population (84%) was more than that between two populations (16%). The isolates, when tested by indirect competitive ELISA, showed about 68.5% of them to be atoxigenic. Composite analysis between the aflatoxin production and AFLP data was found to be ineffective in separating the isolate types by aflatoxigenicity. Certain unique fragments, with respect to individual isolates, were also identified that may be used for development of SCAR marker to aid in rapid and precise identification of isolates.

Copper induction and differential expression of laccase in Aspergillus flavus

Aspergillus flavus was isolated from soil and exhibited laccase activity under both constitutive and copper induced conditions. Spiking the medium with 1 mM copper sulfate resulted in an increase in the activity which reached 51.84 U/mL, a distinctive protein band was detected at 60 kDa. The extracellular enzyme was purified 81 fold using gel filtration chromatography and resulted in two different laccase fractions L1 and L2, the latter had a higher enzymatic activity which reached 79.57 U/mL and specific activity of 64.17 U/μg protein. The analysis of the spectrum of the L2 fraction showed a shoulder at 330 nm which is characteristic for T2/T3 copper centers; both copper and zinc were detected suggesting that this is an unconventional white laccase. Primers of laccase gene were designed and synthesized to recover specific gene from A. flavus. Sequence analysis indicated putative laccase (Genbank ID: JF683612) at the amino acid level suggesting a close identity to laccases from other genera containing the copper binding site. Decolorization of textile waste water under different conditions showed possible application in bioremediation within a short period of time. The effect of copper on A. flavus was concentration dependent.

Aspergillosis of the nose and paranasal sinuses: a review of 54 cases

Aspergillus species are considered opportunistic fungi of increasing clinical importance. Informationregarding extrapulmonary involvement is scarce. The aim of this study was to isolate the differentspecies of Aspergillus from patients with rhinosinusitis. A retrospective study was conducted ina university hospital in Porto Alegre, Brazil (1986–2014). For mycological diagnoses, paranasaltissue obtained at surgery was subjected to histopathology examination and sent for fungal cultures.Of the 54 samples analyzed, 32 were diagnosed positive by culture. The underlying causes ofimmunodeficiency were: six with transplantation (three bone marrow,two lung, one kidney) andtwo with hematological disease (one bone marrow neoplasia and two leukemia). In the presentstudy, the clinical manifestations of rhinosinusitis aspergillosis were: 20 allergic reactions, 20fungus balls, and 14 acute invasive cases. The species isolated from the 54 samples were: Aspergillusfumigatus (n=14); A. flavus (n=6); A. niger (n=2); A. terreus (n=1); A. fischeri (n=1); and Aspergillussp., (n=3). Two concomitant species of Aspergillus were observed in two patients: A. fumigatus andA. flavus; and A. fumigatus and A. niger. In four patients, Aspergillus was associated with other fungi. These were: A. flavus and Fusarium, A. fumigatus and Rhyzopus, A. flavus and Mucorales, and Aspergillus sp. and Mucorales. The most common species of Aspergillus that were responsiblefor paranasal sinus infections were A. fumigatus, A. flavus, and A. niger...

Espécies de Aspergillus são considerados fungos oportunistas de crescente importância clínica.Informações sobre o envolvimento extrapulmonar é escassa. O objetivo deste estudo foi isolaras diferentes espécies de Aspergillus em pacientes com rinossinusite. Um estudo retrospectivofoi realizado em um hospital universitário em Porto Alegre, Brasil (1986-2014). Para diagnósticomicológico, tecido paranasais obtido no momento da cirurgia foi submetido a exame histopatológicoe encaminhados para cultivos de fungos. Das 54 amostras analisadas, 32 foram diagnosticados pelocultivo positivo. As causas subjacentes da imunodeficiência foram: seis com transplante (medulaóssea, três, pulmão, dois; rim, um) e dois com doenças hematológicas (neoplasia osso estreito,um; leucemia, duas). No presente estudo, as manifestações clínicas de rinossinusite aspergilarforam: alérgica, 20; bolas fúngica, 20; e aguda invasiva, 14. As espécies fúngicas isoladas foram:Aspergillus fumigatus, 14; A. flavus, seis; A. niger, dois; A. terreus, um; A. fischeri, um; e Aspergillussp., três. Duas espécies de Aspergillus concomitantes foram observadas em dois pacientes: A.fumigatus e A. flavus; e A. fumigatus e A. niger. Em quatro pacientes, Aspergillus foi associado comoutros fungos: A. flavus e Fusarium, um; A. fumigatus e Rhyzopus, um; A. flavus e Mucorales, um; eAspergillus sp. e Mucorales, um. Os isolados mais comuns de Aspergillus que são responsáveis porinfecções dos seios paranasais são A. fumigatus, A. flavus e A. niger...

Screening the antifungal activity of essential oils against decay fungi from palmyrah leaf handicrafts

BACKGROUND: The whitish tender leaves of Palmyrah are used for making handicrafts. The problem with these articles is discolouration with time and become more brittle due to fungal attack. This could be prevented by some protective coating. Instead of expensive and harmful chemicals we decided to test natural plant essential oils to control fungal attack. Palmyrah leaf article decay fungi were isolated from two different sites of Jaffna peninsula. In this investigation Antifungal Activity of different plant essential oils from neem (Azadirachta indica), castor (Ricinus communis), citronella (Cymbopogon sp) and camphor (Cinnamomum camphora) obtained from local market have been evaluated against isolated fungi. For screening of Antifungal activity, tests and controls were set to determine minimum inhibitory concentration (MIC) and Percentage of Growth Inhibition. RESULTS: Morphologically three different types of Palmyrah leaf decay fungi were isolated and characterized asAspergillus niger, Aspergillus flavus and Penicillium sp. Neem and castor oils have recorded no significant (0.05 > P) antifungal activity while citronella and camphor oils showed significantly different antifungal activity compared with control. Camphor oil and Citronella oil showed 100, 58.13% of average growth inhibition for A. niger. 96.38, 51.32% for A.flavus and 84.99, 72.76% forPenicillium sp respectively. Camphor oil showed highest percentage of growth inhibition at lowest minimum inhibitory concentration compared with citronella oil. Camphor oil was found to be highly antifungal and most effective against A niger, and A. flavus, compared with Penicillium sp and gave 100 percentage of growth inhibitions at 5, 1 and 15 ml/dl minimum inhibitory concentration respectively. CONCLUSION: Significantly higher broad-spectrum of antifungal activity was observed in camphor oil than other tested oils because it showed highest percentage of growth inhibition at lowest inhibitory concentration. Therefore it could be used for the development of new environmental friendly antifungal agent for the preservation of leafy handicrafts. Further formulation, field experiments are necessary to achieve this target.

Efficacy of aqueous garlic extract on growth, aflatoxin B1 production, and Cyto-morphological aberrations of Aspergillus flavus, causing human ophthalmic infection: topical treatment of A. flavus keratitis

By using agar well diffusion assay, antifungal activity of aqueous extract prepared from Egyptian garlic (Allium sativum L.) was evaluated in vitro against two strains of Aspergillus flavus (OC1 and OC10) causing human ocular infection. The recorded minimum inhibitory concentration (MIC) for growth inhibition of both strains was 3.60 mg/ml. Aqueous garlic extract (AGE) was used in successive in vivo tests as an attempt to cure rabbit's fungal keratitis caused by A. flavus OC1. Findings showed that diluted preparation of AGE was effective topical antifungal agent and succeeded to cure severe A. flavus keratitis in a time course less than 10 days without any observable side effects. Microscopic examination showed that AGE induced deleterious cyto-morphological aberrations inA. flavus target cells. AGE applied to Czapek's broth via contact method was more effective on growth, spores and aflatoxin B1 production than AGE applied to the same broth at the same concentration via fumigation method.

Dermatomicosis en ancianos institucionalizados y estudios de sensibilidad in vitro a los antifúngicos sistémicos / Dermatomycosis in institutionalized elderly patients and in vitro sensitivity study to systemic antifungus

Se determinó la prevalencia de dermatomicosis en ancianos institucionalizados de Ciudad Bilívar, Estado Bolívar, Venezuela, y se evaluó la sensibilidad in vitro de los aislamientos clínicos a los antifúngicos itraconazol, fluconazol y terbinafina mediante el método de microdilución en medio líquido, recomendado por el Comité Internacional de Laboratorios Clínicos (M38-P), con algunas modificicaciones. Los hongos fueron identificados mediante métodos tradicionales. Las levaduras se identificaron mediante pruebas bioquímicas, sistema Api 20 C AUX (Biomérieux SA®, France) y crecimiento en medio de Staib. Se estudiaron 74 ancianos, todos recluidos en el Asilo "San Vicente de Paúl" y el Geriátrico "Carlos Fragachán" quienes dieron consentimiento por escrito para participar en el estudio. La edad de los pacientes estuvo comprendida entre 63 y 98 años (80 ± 8,4 años), la mayoría eran hombres (73%). Todos los pacientes tenían lesiones sugestivas de onicomicosis en los pies. El único dermatofito aislado fue Trichophyton rubrum (n=2) el cual resultó sensible al Itraconazol, terbinafina y sensibilidad variable a flucozazol. Asimismo se logró aislar Aspergillus niger (n=5; 6,7%) demostrándose sensible a terbinafina y fluconazol con sensibilidad variable a itraconazol. Candida albicans (n=3; 4,1%) fue sensible a fluconazol, resistentes a itraconazol y variable a la terbinafina. Aspergillus flavus fue aislado en dos casos (2,7%). Además de Geomyces sp, Fusarium oxysporum y Pseudeurotium ovale. Se concluye que existe una prevalencia baja de dermatomicosis en los ancianos institucionalizados de Ciudad Bolívar y que las lesiones clinicamente observadas son debidas a los cambios degenerativos propios de la edad.

A study determine prevalence of dermatomycosis in 74 institutionalized elderly patients was conductted in Ciudad Bolivar, state of Bolivar, Venezuela. Clinical isolates were assayed for in vitro sensitivity to itraconazole, fluconazole, and terbinafine using a slightly modified version of the microdilution method in liquid medium recommended by the International Committee of Clinical Laboratory (M38-P). Traditional methods were used to identify the fungi. The yeasts were identified by Api 20C AUX biochemical testing (bioMérieux SA®, France) and growth on Staib media. The elders, mostly men (73%), from the "San Vicente de Paúl" Nursing Home and the "Carlos Fragachan" Geriatric Hospital, were aged between 63 and 98 (80 ± 8.4 years). All the patients, whose written consent was secured, had lesions suggestive of onychomycosis. Trichophyton rubrum was the only isolated dermatophyte (n=2), which resulted sensitive to itraconazole and terbinafine, with variable sensitivity to fluconazole. Aspergillus niger (n=5;6.7%) was sensitive to terbinafine and fluconazole with variable itraconazole sensitivity. Candida albicans (n=3; 4.1%) was fluconazole sensitive, resistant to itraconazole, and variable to terbinafine. Aspergillus flavus was isolated in two cases 2.7%). Geomyces sp., Fusarium oxysporum, and Pseudeurotium ovale were also isolated. It is concluded that there is a low prevalence of dermatomycosis among institutionalized elders in Ciudad Bolivar, and that the lesions clinically observed were due to degenerative changes naturally occurring with aging.

Ação fungitóxica do óleo essencial de Tanaecium nocturnum (Barb. Rodr.) Bur. e K. Shum sobre o Aspergillus flavus isolado da castanha-do-Brasil (Bertholletia excelsa) / Fungitoxic action of the essential oil of Tanaecium nocturnum (Barb. Rodr.) Bur. and K. Shum on Aspergillus flavus isolated from the Brazil nut (Bertholletia excelsa)

Neste trabalho, avaliou-se a capacidade fungitóxica do óleo essencial de folhas frescas de Tanaecium nocturnum sobre o Aspergillus flavus isolado da castanha-do-brasil, por meio das técnicas de contato e fumigação. Pelos resultados dos bioensaios realizados até 10 dias de incubação, verificou-se que a inibição total do crescimento micelial ocorreu quando se utilizou o óleo essencial nas concentrações de 782 ppm (técnica de contato) e 1000 ppm (técnica de fumigação). Em ambas as técnicas, o óleo essencial inibiu a esporulação a partir da concentração de 500 ppm. Observou-se que nos cinco primeiros dias de incubação não houve diferença significativa nos resultados apresentados pelas duas técnicas estudadas, havendo a partir daí uma redução da atividade do óleo essencial nas concentrações inferiores a 1000 ppm pelo teste de fumigação. A ação fungitóxica do óleo essencial sobre o microrganismo estudado pode ser atribuída à presença do benzaldeído (composto majoritário do óleo essencial estudado), em associação com outros compostos também presentes nesse óleo essencial, tais como; álcool benzílico, benzoato de benzila e mandelonitrila.

The present work sought to evaluate the fungitoxic activity of the essential oil from fresh Tanaecium nocturnum fresh leaves on Aspergillus flavus isolated from Brazil nuts, using contact and fumigation techniques. The results of bioassays performed up to 10 days of incubation demonstrated that total inhibition of mycelial growth occurred when using the essential oil at concentrations of 782 ppm (contact technique) and 1000 ppm (fumigation technique). In both techniques, the essential oil inhibited the formation of spores at the concentration of 500 ppm. No significant difference in the results presented by the two techniques was observed in the first five days of incubation. After this period, the essential oil showed a reduction in activity at concentrations lower than 1000 ppm in the fumigation test. The fungitoxic activity of the essential oil on the organism studied can be attributed to the presence of benzaldehyde (major component of the essential oil), in combination with other compounds also present in this oil, such as, benzyl alcohol, benzyl benzoate and mandelonitrila.

Nutritional changes in powdered red pepper upon in vitro infection of Aspergillus flavus / Alterações nutricionais em pimenta vermelha em pó após infecção in vitro com Aspergillus flavus

Quantitative losses in various biochemical constituents like capsaicin, carotenes, ascorbic acid, polyphenols,mineral matter, sugars (soluble and insoluble), protein and fat were estimated after the successful growth ofAspergillus flavus for 30 days on powdered red pepper. The fungal biomass was measured by ergosterolcontent and Aflatoxin B1 by HPLC. Amongst the various nutritional constituents evaluated for nutritionallosses and changes the highest nutritional loss was reported in total carotenoids (88.55%) followed by totalsugars (85.5%). The protein content of the infected sample increased from 18.01% to 23%. The nutritional profile of chilli powder (Capsicum annum var. sannam L.) shows highest share of total soluble sugars (32.89%) and fiber content (21.05%), followed by protein (18.01%) and fat (13.32%) making it an ideal solid - substrate for mould growth. At the end of incubation the fungal biomass was 192. 25 mg / 100 gram powder, total plate count 17.5 X 10 4 CFU/g and Aflatoxin B1 content was 30.06 μg / kg.

Foram avaliadas as perdas de vários constituintes bioquímicos como capsaicina, carotenos, acido ascórbico,polifenóis, matéria orgânica, açucares (solúveis e insolúveis), proteína e gordura em pimenta vermelha em pó após a multiplicação de Aspergillus flavus por 30 dias. A biomassa fúngica foi mensurada pelo conteúdo de ergosterol e aflatoxina por HPLC. Entre os vários constituintes avaliados, a maiorperda foi a de carotenóides totais (88,55%), seguido de açucares totais (85,5%). O conteúdo protéico da amostra infectada aumentou de 18,01% para 23%. O perfil nutricional da pimenta em pó (Capsicum annum var. sannam L.) indica alto teor de açucares totais (32,89%) e fibras (21,05%), seguido de proteína (18,01%) e gordura (13,32%), tornando-a umsubstrato ideal para crescimento de fungos. Ao final dos 30 dias, a biomassa fúngica foi 192,25 mg/100g, a contagem total em placas foi 17,5 x 104 CFU/g e o conteúdo de aflatoxina B1foi 30,06 μg/kg.

Invasive Aspergillus flavus sinusitis: case report in a patient with biphenotypic acute leukemia / Sinusite invasiva por Aspergillus flavus: relato de um caso associado a leucemia aguda bifenotípica

Here we report a case of invasive pansinusitis with proptosis of the right eye caused by Aspergillus flavus in an immunocompromised patient with acute biphenotypic leukemia without aggressive therapy response.

Descreve-se um caso de pansinusite invasiva com proptose do globo ocular direito causado por Aspergillus flavus em um paciente imunossuprimido com leucemia aguda bifenotípica sem resposta a terapia agressiva.

Genetic diversity of environmental Aspergillus flavus strains in the state of São Paulo, Brazil by random amplified polymorphic DNA

Aspergillus flavus is a very important toxigenic fungus that produces aflatoxins, a group of extremely toxic substances to man and animals. Toxigenic fungi can grow in feed crops, such as maize, peanuts, and soybeans, being thus of high concern for public health. There are toxigenic and non-toxigenic A. flavus variants, but the necessary conditions for expressing the toxigenic potential are not fully understood. Therefore, we have studied total-DNA polymorphism from toxigenic and non toxigenic A. flavus strains isolated from maize crops and soil at two geographic locations, 300 km apart, in the Southeast region of Brazil. Total DNA from each A. flavus isolate was extracted and subjected to polymerase chain reaction amplification with five randomic primers through the RAPD (random amplified polymorphic DNA) technique. Phenetic and cladistic analyses of the data, based on bootstrap analyses, led us to conclude that RAPD was not suitable to discriminate toxigenic from non toxigenic strains. But the present results support the use of RAPD for strain characterization, especially for preliminary evaluation over extensive collections.

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...

Prevalence of Aspergillus species in clinical samples isolated in an Indian tertiary care hospital.

CONTEXT (BACKGROUND): In recent times, it has become important to determine the prevalence of different Aspergillus species in clinical samples in view of difference in antifungal susceptibility noted in some species. AIMS: To determine the species prevalence of Aspergillus isolates in various clinical samples received in the Mycology Laboratory at our institute. METHOD: Over a period of 4-years, a total of 18,731 samples were processed, and species identification carried out by standard microbiological methods. RESULTS: Four hundred and fifty six samples (2.43%) were culture positive for Aspergillus species. A.flavus (46.93%) was the most common isolate, followed by A.fumigatus (37.72%) and A.niger (15.35%). It was observed that A.fumigatus was the predominant species isolated from blood and respiratory specimens, A.flavus was predominantly isolated from nasal polyps whereas A.niger predominated in nail specimens. Culture positivity was highest in the age group 12-65 years and in males. Sixty-nine patients (15.13%) were admitted to the intensive care unit. CONCLUSIONS: The study highlights the diverse manifestations caused by Aspergillus species in human beings and also throws light on the different species prevalent locally. The knowledge would prove useful in selecting empirical antifungal therapy and formulating prophylactic and pre-emptive strategies.