Invasive aspergillosis in a user of inhaled cocaine: rhinosinusitis with bone and cartilage destruction

Aspergillosis is an infection caused by saprophytic fungi of the genus Aspergillus, which typically occurs in immunosuppressed individuals, but has also been reported in immunocompetent patients. The main routes of entry are the respiratory tract, skin, cornea, and ear, and the infection may be localized or disseminated by contiguity or vascular invasion. We report a severe case of rhinosinusitis with cutaneous involvement, caused by invasive aspergillosis, in an immunocompetent user of inhaled cocaine. Invasive aspergillosis related to cocaine abuse has not yet been reported in the literature. After itraconazole treatment and surgical debridement, complete clinical remission was achieved. Nasal reconstruction with a skin graft over a silicone prosthesis resulted in a satisfactory esthetic outcome.

Fungos contaminantes e produtores de aflatoxinas em castanha do Pará (Bertholletia excelsa HUMB. & BONPL 1808)

Cem amostras de castanhas do Para, procedentes dos Estados do Amazonas e Sao Paulo, foram analisadas micológico e toxicologicamente para aflatoxinas B1 e G1. Isolou-se 312 colônias de fungos, sendo 91 do gênero Aspergillus, 83 do gênero Penicillium e as restantes incluídas em 23 gêneros diferentes. Entre as amostras de Aspergillus, 26 foram aflatoxigênicos, distribuídos em 3 espécies: Aspergillus flavas Link (18); Aspergillus parasiticus Speare (7) e Aspergillus fresenii Subram (1). Dos 100 extratos de castanhas analisados 3 foram positivos para aflatoxina.

Seleção de fungos produtores de β-D-frutosiltransferase por fermentação em estado sólido

A enzima β-D-frutosiltransferase é responsável pela síntese de FOS (frutooligossacarídeos) a partir de sacarose por reação de transfrutosilação é produzida por diferentes micro-organismos, principalmente por fungos filamentosos. O objetivo deste trabalho foi selecionar a melhor linhagem fúngica produtora da -D-frutosiltransferase por fermentação em estado sólido, bem como o método de extração. A fermentação em estado sólido utilizando o substrato farelo de trigo umedecido com solução de sacarose atingindo 70% de umidade na concentração de esporos de 107 no tempo de 96 horas de crescimento. Todas as linhagens manipuladas apresentaram atividade hidrolítica, no entanto apenas uma linhagem não demonstrou atividade transfrutosilação. O isolado SIS 14 que pertence ao gênero Aspergillus sp. destacou-se pelos maiores valores em atividade no método de extração utilizando água destilada, apresentando 300,90 U/mL na atividade de transfrutosilação e na atividade hidrolítica de 155,74 U/mL. Contudo, pode-se perceber que dos solventes estudados a água destilada foi melhor obtendo o valor em atividade de transfrutosilação, como também a linhagem SIS 14 é promissora para a produção da β-D-frutosiltransferase.

Atividade antimicrobiana de fungos endofíticos isolados de plantas tóxicas da amazônia: Palicourea longiflora (aubl.) rich e Strychnos cogens bentham

Das plantas tóxicas da Amazônia Palicourea longiflora e Strychnos cogens foram isolados 571 fungos endofíticos e 74 bactérias endofíticas. Palicourea longiflora (Rubiaceae) e outras espécies desse gênero estão relacionadas a 90% das mortes de gado na região Amazônica. Strychnos cogens (Loganiaceae) e outras espécies de Strychnos são utilizadas pelos indígenas na confecção de "curares". Entre os endófitos isolados de P. longiflora foram identificados os fungos: Colletotrichum sp. e seu telemorfo Glomerella sp., Guignardia sp., Aspergillus niger, Phomopsis sp. e Xylaria sp., além da bactéria Burkholderia gladioli, pertencente a um grupo de fixadoras de nitrogênio. Dos isolados de S. cogens foram identificados os fungos: Colletotrichum sp., Guignardia sp., Aspergillus niger e Trichoderma sp. Uma amostra de 79 isolados fúngicos teve seus metabólitos extracelulares bioensaiados contra microrganismos patogênicos e fitopatogênicos: 19 isolados inibiram um ou mais microrganismos-teste. Dos oito isolados com melhores resultados de inibição, as móleculas bioativas eram menores que 12.000 daltons, fato verificado pela diálise dos metabólitos.

Aspergilli and Penicillia related to the surface of ripening italian grana type cheese and its ripening environment

Information available on the mycoflora associated to ripening Italian “grana type” cheese is very poor. Recently, ochratoxin A (OTA) was detected in samples of packed grated cheese [1]; therefore, the need of information to perform a risk management was highlighted. Moreover, sterigmatocystin (STC) has been reported in cheese and it is considered an emerging problem. Despite the fact that both of them are mycotoxins included in group 2B by IARC [2,3], no European regulation exists. So, the main goal of this work is to give for the first time a general overview about Penicillia and Aspergilli growing on the surface of ripening “grana type” cheese, with particular attention on mycotoxigenic species. To perform this, in 2013 and 2014 crust samples were scratched from ripening grana cheese wheels and also Potato Dextrose Agar plates were exposed to monitor ripening house air. Then, 140 fungal isolates were randomly chosen, purified and monosporic colonies were obtained for their identification at specie level. A polyphasic approach is followed, based on morphological characterisation, toxic extrolites profiling and gene sequencing. The identification is still in progress, but the first results based on the morphological approach showed the presence of mycotoxigenic Aspergilli (Aspergillus flavus and A. versicolor) and various Penicillium species; among them Penicillium chrysogenum, P. implicatum and P. solitum were identified. Only P. chrysogenum was reported to produce the mycotoxins cyclopiazonic acid (CPA) and roquefortine-C (ROQ-C) [4]. These results will be presented and discussed. [1] A. Biancardi, R. Piro, G. Galaverna, C. Dall’Asta, "A simple and reliable liquid chromatography–tandem mass spectrometry method for determination of ochratoxin A in hard cheese" International Journal of Food Sciences and Nutrition 64 (5), 2013, 632 – 640. [2] International Agency for Research on Cancer (IARC) “IARC Monographs on the Evaluation of Carcinogenic Risks to Humans” 31, 1983, 191 – 199. [3] International Agency for Research on Cancer (IARC) “IARC Monographs on the Evaluation of carcinogenic Risks to Humans”, suppl. 7, 1987, 72. [4] J. I. Pitt, D. A. Hocking, “Fungi and Food Spoilage” 1997, 291.

Antimicrobial activity of bergenin from Endopleura uchi (Huber) Cuatrec

Endopleura uchi (Huber) Cuatrec. is an Amazon species traditionally used as treatment for inflammations and female disorders. Bergenin was isolated from ethyl acetate fraction of bark of E. uchi by using column chromatography over sephadex LH-20 and then silica gel 60 flash. Its structure was identified on the basis of its NMR spectra. The antimicrobial activity of bergenin and fractions of methanol extract of E. uchi were evaluated against ATCC microorganisms (Escherichia coli, Salmonella enteritidis, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus, Candida albicans, C. guilliermondii, Aspergillus flavus, A. nidulans). Clinically isolated strains of all of these microorganisms, along with C. tropicalis, A. niger, Shigella sonnei, Serratia marcenses and Klebsiella pneumoniae were also evaluated. The growth inhibition caused by bergenin, extracts and fractions of E. uchi against ATCC microorganisms were similar to the inhibition to microorganisms clinically isolated. The ethyl acetate fraction and the isolate bergenin inhibit the growth of the yeasts C. albicans, C. tropicalis, and C. guilliermondii, but present lower activity against filamentous fungi Aspergillus flavus, A. nidulans, A. niger, and did not inhibit the Gram positive and Gram negative bacteria. The activity of the ethyl acetate fraction and bergenin are in agreement wit its high concentration found in bark extract of E. uchi. Moreover, the selective activity against three Candida species helps to understand its traditional use against infections that affect women.

Superação de dormência na qualidade de sementes e mudas: influência na produção de Senna multijuga (L. C. Rich.) Irwin & Barneby

O objetivo deste trabalho foi determinar a qualidade fisiológica e sanitária de sementes de Senna multijuga(L. C. Rich.) Irwin & Barneby relacionada aos métodos de superação de dormência e à interferência na produção de mudas. As sementes foram submetidas aos seguintes métodos: imersão em água fervente, as sementes foram imersas em água, com temperatura de 100°C, até esfriar, por 24 horas; escarificação ácida, onde as sementes foram imersas em ácido sulfúrico (H2SO4) a 90%, por 10 e 20 minutos, e testemunha (sem tratamento). Foram realizados os testes de sanidade, germinação, tetrazólio e avaliação da qualidade das mudas. O delineamento experimental foi inteiramente casualizado. Para a avaliação da germinação foi utilizado um esquema fatorial (4 X 2), com quatro métodos de superação de dormência X dois fotoperíodos, para os substratos rolo-de-papel e vermiculita. A escarificação ácida constituiu-se no método mais eficiente para a superação da dormência das sementes de Senna multijuga. Penicillium sp. e Aspergillus sp. tiveram sua incidência aumentada quando o tegumento foi danificado pela escarificação ácida por 20 minutos. O controle de Fusarium spp. aumentou gradativamente com o aumento do tempo de exposição ao ácido sulfúrico.

A review of mycotoxins in food and feed products in Portugal and estimation of probable daily intakes

Mycotoxins are toxic secondary metabolites produced by filamentous fungi that occur naturally in agricultural commodities worldwide. Aflatoxins, ochratoxin A, patulin, fumonisins, zearalenone, trichothecenes and ergot alkaloids are presently the most important for food and feed safety. These compounds are produced by several species that belong to the Aspergillus, Penicillium, Fusarium and Claviceps genera and can be carcinogenic, mutagenic, teratogenic, cytotoxic, neurotoxic, nephrotoxic, estrogenic and immunosuppressant. Human and animal exposure to mycotoxins is generally assessed by taking into account data on the occurrence of mycotoxins in food and feed as well as data on the consumption patterns of the concerned population. This evaluation is crucial to support measures to reduce consumer exposure to mycotoxins. This work reviews the occurrence and levels of mycotoxins in Portuguese food and feed to provide a global overview of this issue in Portugal. With the information collected, the exposure of the Portuguese population to those mycotoxins is assessed, and the estimated dietary intakes are presented.

Qualidade da castanha-do-brasil do comércio de Rio Branco, Acre

Este trabalho teve como objetivo avaliar a qualidade de castanhas-do-brasil beneficiadas e comercializadas em Rio Branco, Acre. Foram analisadas amostras das três marcas de castanha encontradas no mercado local quanto às variáveis: atividade de água, teor de umidade, contagem total de fungos filamentosos, quantificação de Aspergillus flavus e de A. parasiticus, bem como quantificação de aflatoxinas B1, B2, G1 e G2. As castanhas do comércio se encontravam com um teor de umidade e atividade de água adequados, o que pode ter sido responsável pela baixa contaminação por fungos e por aflatoxinas. Quanto a estas micotoxinas, as amostras estão de acordo com o recomendado pela Anvisa, podendo ser esta uma consequência da grande divulgação no Estado do uso de Boas Práticas no manejo da castanha.

Laccase-catalyzed synthesis of conducting polyaniline-lignosulfonate composite

Enzymatic polymerization of aniline was first performed in lignosulfonate (LGS) template system. High-redox-potential catalyst laccase, isolated from Aspergillus, was used as a biocatalyst in the synthesis of conducting polyaniline/lignosulfonate (PANI-ES-LGS) complex using atmospheric oxygen as the oxidizing agent. The linear templates (LGS), also serving as the dopants, could facilitate the directional alignment of the monomer and improve the solubility of the conducting polymer. The process of the polymerization was monitored using UV-Vis spectroscopy, by which the conditions for laccase-catalyzed synthesis of PANI-ES-LGS complex were also optimized. The structure characterizations and solubility of the complex were carried out using corresponding characterization techniques respectively. The PANI-ES-LGS suspensions obtained was used as coating for cotton with a conventional padder to explore the applications of the complex. The variable optoelectronic properties of the coated cotton were confirmed by cyclic voltammetry and color strength test. The molecular weight changes of LGS treated by laccase were also studied to discuss the mechanism of laccase catalyzed aniline polymerization in LGS template system.

Citric acid production by Y. lipolytica W29 from crude glycerol: strain improvement by mutagenesis

[Excerpt] Citric acid, an important and versatile organic acid extensively used in several industries, is originally produced by Aspergillus niger in submerged fermentation from molasses [1]. However, Yarrowia lipolytica have been studied and demonstrate a great potential as citric acid producer from several carbon sources [1–5] including crude glycerol, a low cost byproduct from the biodiesel industry [6]. The simultaneous production of the isomer isocitric acid is the major problem in using this yeast in the citric acid production. (...)

Aflatoxins detoxification by gamma irradiation

[Excerpt] Mycotoxins are secondary toxic metabolites of filamentous fungi. Aflatoxins (AFs) are produced to Aspergillus species such as A. flavus and A. parasiticus. These fungi are ubiquitous in nature and usually found on agricultural commodities. Therefore, AFs are encountered in many important foodstuff, including wheat, rice, maize, peanuts, sorghum, pearl millet, spices, oilseeds, tree nuts and milk. Due to the high toxicity of AFs, many methods have been studied to reduce or eliminate these mycotoxins from food and feed. Gamma irradiation is one technology that has been investigated with promising results. The aims of this study were (I) to study the effect of gamma radiation on aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatoxin G2 (II) to evaluate the effect of the presence of water on AFs degradation during the irradiation process; and (IV) to evaluate the cytotoxicity of radiolytic products formed. (...)

Current situation on the occurrence of mycotoxins and toxigenic fungi in Portugal

Mycotoxins are secondary metabolites produced by filamentous fungi that are toxic for humans and animals in small amounts and that are found worldwide in a large number of agricultural commodities. They are usually ingested involuntarily, when contaminated plant products are consumed, and represent a great risk for public health. Therefore, governments throughout the world have imposed strict legal limits for their levels in food and feed products in order to reduce potential health risks for consumers. Despite of its ubiquity, the mycotoxin problem is mainly dependent on regional factors, such as the mycotoxigenic characteristics of the local mycoflora, the local climate conditions, and the local agricultural practices. For this reason, a constant vigilance from local governmental food safety agencies and from the local researcher community is needed. This communication will review the current situation on the occurrence of mycotoxigenic fungi in some Portuguese cultures, such as wine grapes, corn and dried fruits. Particular attention will be given to the incidence of mycotoxigenic Aspergillus strains in those cultures and to the levels of ochratoxin A, aflatoxins, cyclopiazonic acid and fumonisin B2 produced. Data will be discussed taking into account the geographical origin of the isolates and the particular climate conditions of each sampling region. An updated review on the levels of the main mycotoxins found in local products and in imported commodities will also be presented.

Simultaneous detection of cyclopiazonic acid and aflatoxin B1 by HPLC in methanol/water mobile phase

A simple procedure for the simultaneous detection of cyclopiazonic acid (CPA) and aflatoxin B1 from fungal extracts is presented, using a methanol and water mobile phase and fluorescence detection. This methodology has been tested with standard solutions of both mycotoxins CPA and Aflatoxin B1 and with methanolic extracts of Aspergillus section Flavi strains, previously characterized for their mycotoxin production profile. Previously available methodology required the use of two different chromatographic runs for these mycotoxins, with distinct columns and detectors (fluorescence detection with a post-column photochemical derivatization (PHRED) for aflatoxin B1 and UV detection for CPA). The proposed method detects both mycotoxins in a single run. Data from these assays will be presented and discussed.

Biosynthesis, detection and toxicology of ochratoxins - optimisation of production

Ochratoxin A (OTA) is a very well known mycotoxin found in several food commodities for which maximum limits are being discussed in EC in other to produce appropriate regulations. OTA is one of several ochratoxins produced by Aspergillus and Penicillium species. All the compounds in this group have a molecular structure very similar to OTA and some were already isolated from natural substrates. Several of these compounds such as ochratoxin , methyl and ethyl ester of ochratoxin A, 4-R and S-hydroxyochratoxin A, 10-hydroxyochratoxin A and ochratoxin A open lactone are commercially unavailable. However, they can be easily synthesized through OTA modification. With the main objective of its application on further research works, OTA production, isolation and purification has been optimised from an A. alliaceus strain grown on wheat medium. Synthesis and purification of some OTA derivatives has been achieved and an HPLC method for their detection was optimised. Data about their production by several species of Aspergillus will be presented. The toxicological properties of ochratoxins are still not very clear and a future EC safety limit for OTA will depend on e.g., a better clarification of its carcinogenity. Could OTA derivatives play a role here?