Controle químico de doenças foliares na cultura do arroz irrigado

O objetivo deste trabalho foi avaliar a interferência das doenças foliares na produtividade de duas cultivares de arroz irrigado, bem como determinar a interação entre ingredientes ativos e momento de aplicação, para o controle de escaldadura e mancha-marrom. Foram utilizados: tiofanato metílico, tebuconazole, azoxystrobin e trifloxystrobin + propiconazole, aplicados em cinco épocas, às cultivares IRGA 417 e El Passo L 144. Os parâmetros avaliados foram: severidade das doenças e rendimento de grãos. O controle químico proporcionou redução significativa na severidade das doenças, em ambas as cultivares, com reflexo no rendimento de grãos, e foi influenciado pelo produto e pela época de aplicação.

Solubilização do fosfato de ferro em meio de cultura

O objetivo deste trabalho foi avaliar a eficiência de solubilização de FePO4 por Aspergillus niger, em meio de cultura com diferentes fontes de carbono (C) e nitrogênio (N) e com concentrações crescentes de fosfato. A quantidade de fosfato solúvel, acidez e pH final foram determinados após o crescimento do fungo, em cultura estacionária a 30ºC. A eficiência de solubilização aumentou conforme o crescimento do fungo, atingiu o máximo no 11º dia (68%) e depois regrediu. Das fontes de C e de N testadas, as maiores eficiências de solubilização foram obtidas com manitol (21%) e ácido glutâmico (17%). Com o aumento da concentração de fosfato (0 a 1.330 µg mL-1), a máxima eficiência de solubilização (70%) foi obtida com 330 µg mL-1 de PO4(3-) e decresceu até 47% com 1.330 µg mL-1. A produção de ácidos foi o principal mecanismo de solubilização do FePO4, com base na correlação positiva e significativa entre a produção de fosfato e a acidez.

Aplicação do silício para aumentar a resistência do arroz à mancha-parda

O objetivo deste trabalho foi avaliar fontes e métodos de aplicação de silício para aumentar sua absorção e a resistência de plantas de arroz (Oryza sativa), cultivar Metica-1, à mancha-parda (Bipolaris oryzae). Foram conduzidos dois experimentos em casa de vegetação, com solo Latossolo Vermelho. O delineamento experimental foi o completamente casualizado, com oito repetições. As fontes foram wollastonita (silicato de cálcio), aplicada via solo, e silicato de potássio e ácido monossilícico, aplicados via foliar. A aplicação de silício via solo resultou em aumento do teor foliar deste elemento e foi eficiente na indução de resistência à mancha-parda, diferentemente do observado com sua aplicação via foliar.

Review of CT features of saprophytic aspergillosis (Aspergilloma) and correlation with final diagnosis : P21

Purpose: 1. To review Ct features suggestive of saprophytic aspergillosis (aspergilloma) and to correlate them with the final pathological results. 2. To illustrate the wide range of differential diagnosis. Methods and materials: The electronic database of our department from 1995 to 2007 revealed CT reports of 48 patients that had been considered very suggestive of aspergilloma. Two radiologists with 6 and 12 years experience in thoracic radiology jointly reviewed the corresponding CT features including ancillary findings and the underlying lung diseases and correlated them with the final pathological diagnosis. Results: Forty patients could be included in the study (12 women, mean age 52), while in 8 patients there was no adequate clinical follow-up. In 17 patients the diagnosis "mycetoma" due to aspergillus fumigatus infection was confirmed, either by surgery, biopsy or bronchoscopy. In 23 patients, differential diagnoses were found, such as cavitating bronchial carcinoma (n = 7), bacterial abscess (n = 3), typical (n = 2) and atypical (n = 2) tuberculosis, as well as inflammatory changes due to mucoviscidosis (n = 1), Wegener's disease (n = 1) or chronic obstructive pulmonary disease (n = 3). Fibromyxoide hamartoma, lung infarction and bronchomucocele were responsible for the typical CT feature in one patient each. Conclusion: 1. The typical CT feature suggesting mycetoma is softtissue proliferation within a pre-existing wall-thickened lung cavity, oten even considered "pathognomonic". However, this diagnosis was finally confirmed by surgery or laboratory findings in less than 50% of patients only. 2. Since differential diagnoses are very large, not only including cavitating lung cancer and tuberculosis, the individual underlying lung disease needs strongly being taken into account often giving the best clue for the correct diagnosis.

Atividade celulolítica de fungos isolados de bagaço de cana-de-açúcar e madeira em decomposição

O objetivo deste trabalho foi identificar isolados de fungos a partir de bagaço de cana-de-açúcar e madeira em decomposição e avaliar a sua atividade celulolítica em bagaço de cana. Cinco isolados foram avaliados, tendo-se como referências os fungos Trichoderma reesei QM9414 e T. reesei RUT C30. A atividade celulolítica foi estimada pela capacidade hidrolítica do extrato enzimático dos fungos cultivados em bagaço de cana sobre os substratos papel de filtro (atividade celulolítica total) e carboximetilcelulose sódica (atividade da endoglucanase). Os isolados foram identificados pela análise molecular da região 26S rDNA. Os gêneros Paecilomyces, Aspergillus, Acremonium/Penicillium e Trichoderma foram identificados. Embora T. reesei QM9414 tenha apresentado a mais alta atividade celulolítica total, alguns isolados também apresentaram alta atividade de endoglucanase. A biodiversidade, em nichos como bagaço de cana-de-açúcar, pode fornecer linhagens de fungos celulolíticos com grande potencial biotecnológico.

Biocontrol of seed pathogens and growth promotion of common bean seedlings by Trichoderma harzianum

The objective of this work was to evaluate isolates of Trichoderma harzianum regarding biocontrol of common bean seed-borne pathogens, plant growth promotion, and rhizosphere competence. Five isolates of T. harzianum were evaluated and compared with commercial isolate (Ecotrich), Carboxin+Thiram, and an absolute control. Bean seeds of the cultivar Jalo Precoce, contaminated with Aspergillus, Cladosporium, and Sclerotinia sclerotiorum, were microbiolized with antagonists, and seed health tests were carried out. Isolates were evaluated on autoclaved substrate and in field conditions. Ten days after sowing (DAS), plant length was measured. To test rhizosphere competence, isolates were applied in boxes containing autoclaved washed sand, and root colonization was evaluated at 10 DAS, using five plants per box. The most effective isolates in the seed health tests were: CEN287 and CEN289 to control Aspergillus; the commercial isolate to control Cladosporium; and CEN287 and CEN316 to control S. sclerotiorum. Isolates CEN289 and CEN290 promoted bean growth in greenhouse and field. Seed treatment with T. harzianum reduces the incidence of Aspergillus, Cladosporium, and S. sclerotiorum in 'Jalo Precoce' common bean seeds.

Identification of novel secreted proteases during extracellular proteolysis by dermatophytes at acidic pH.

The dermatophytes are a group of closely related fungi which are responsible for the great majority of superficial mycoses in humans and animals. Among various potential virulence factors, their secreted proteolytic activity attracts a lot of attention. Most dermatophyte-secreted proteases which have so far been isolated in vitro are neutral or alkaline enzymes. However, inspection of the recently decoded dermatophyte genomes revealed many other hypothetical secreted proteases, in particular acidic proteases similar to those characterized in Aspergillus spp. The validation of such genome predictions instigated the present study on two dermatophyte species, Microsporum canis and Arthroderma benhamiae. Both fungi were found to grow well in a protein medium at acidic pH, accompanied by extracellular proteolysis. Shotgun MS analysis of secreted protein revealed fundamentally different protease profiles during fungal growth in acidic versus neutral pH conditions. Most notably, novel dermatophyte-secreted proteases were identified at acidic pH such as pepsins, sedolisins and acidic carboxypeptidases. Therefore, our results not only support genome predictions, but demonstrate for the first time the secretion of acidic proteases by dermatophytes. Our findings also suggest the existence of different pathways of protein degradation into amino acids and short peptides in these highly specialized pathogenic fungi.

Trichophyton rubrum secreted and membrane-associated carboxypeptidases

Dermatophytes are the most common agents of superficial mycoses, and exclusively infect stratum corneum, nails or hair. Therefore, secreted proteolytic activity is considered a virulence trait of these fungi. In a medium containing protein as a sole nitrogen and carbon source Trichophyton rubrum secretes a metallocarboxypeptidase (TruMcpA) of the M14 family according to the MEROPS proteolytic enzyme database. TruMcpA is homologous to human pancreatic carboxypeptidase A, and is synthesized as a precursor in a preproprotein form. The propeptide is removed to generate the mature active enzyme alternatively by either one of two subtilisins which are concomitantly secreted by the fungus. In addition, T. rubrum was shown to possess two genes (TruSCPA and TruSCPB) encoding serine carboxypeptidases of the S10 family which are homologues of the previously characterized Aspergillus and Penicillium secreted acid carboxypeptidases. However, in contrast to the Aspergillus and Penicillium homologues, TruScpA and TruScpB enzymes are not secreted into the environment, but are membrane-associated with a glycosylphosphatidylinositol (GPI) anchor. During infection, T. rubrum secreted and GPI-anchored carboxypeptidases may contribute to fungal virulence by cooperating with previously characterized endoproteases and aminopeptidases in the degradation of compact keratinized tissues into assimilable amino acids and short peptides.

Malassezia yeasts activate the NLRP3 inflammasome in antigen-presenting cells via Syk-kinase signalling.

Although being a normal part of the skin flora, yeasts of the genus Malassezia are associated with several common dermatologic conditions including pityriasis versicolour, seborrhoeic dermatitis (SD), folliculitis, atopic eczema/dermatitis (AE/AD) and dandruff. While Malassezia spp. are aetiological agents of pityriasis versicolour, a causal role of Malassezia spp. in AE/AD and SD remains to be established. Previous reports have shown that fungi such as Candida albicans and Aspergillus fumigatus are able to efficiently activate the NLRP3 inflammasome leading to robust secretion of the pro-inflammatory cytokine IL-1β. To date, innate immune responses to Malassezia spp. are not well characterized. Here, we show that different Malassezia species could induce NLRP3 inflammasome activation and subsequent IL-1β secretion in human antigen-presenting cells. In contrast, keratinocytes were not able to secrete IL-1β when exposed to Malassezia spp. Moreover, we demonstrate that IL-1β secretion in antigen-presenting cells was dependent on Syk-kinase signalling. Our results identify Malassezia spp. as potential strong inducers of pro-inflammatory responses when taken up by antigen-presenting cells and identify C-type lectin receptors and the NLRP3 inflammasome as crucial actors in this process.

Qualidade fisiológica e sanitária de sementes de amendoim durante o processo de produção

O objetivo deste trabalho foi avaliar o efeito das etapas do processo de produção sobre a qualidade fisiológica e sanitária de sementes de amendoim. Utilizou-se o delineamento experimental inteiramente casualizado com 12 tratamentos e quatro repetições. Os tratamentos consistiram de arranquio das plantas, recolha, transporte, secagem, armazenamento (dois, quatro e seis meses), além das seguintes etapas de beneficiamento: trilha mecânica, classificação por tamanho, separação por densidade e coloração, e tratamento químico. Avaliaram-se: o teor de água, a qualidade sanitária, a germinação e o vigor de sementes, após cada tratamento. Aspergillus spp. e Penicillium sp. foram encontrados nas sementes. No primeiro ano, após a trilha mecânica, as sementes apresentaram baixo desempenho. No processo de produção, o armazenamento promoveu a contaminação de 100% das sementes por Aspergillus spp. O tratamento químico foi eficiente na recuperação da qualidade sanitária da semente. As etapas de trilha mecânica e armazenamento reduzem a qualidade fisiológica das sementes de amendoim.

Análise de trilha da contaminação por aflatoxinas em grãos de milho armazenados

O objetivo deste trabalho foi determinar a magnitude dos efeitos diretos e indiretos de parâmetros físico-químicos e microbiológicos sobre a ocorrência de aflatoxinas em grãos de milho armazenados, por meio da análise de trilha. Os dados foram coletados em dois experimentos: um cultivo de inverno (colheita do milho em julho de 2012) e um cultivo de verão (colheita em fevereiro de 2013). Os grãos foram armazenados por 12 meses, em dois tipos de acondicionamentos: em sacarias convencionais de polipropileno e a granel em silo metálico. Avaliaram-se 14 características dos grãos. Calculou-se a matriz de correlações de Pearson entre todos os parâmetros, realizou-se o diagnóstico de multicolinearidade e, posteriormente, aplicou-se a análise de trilha com regressão em crista. Observou-se forte influência das condições ambientais entre as safras de inverno e de verão, e entre época e tipo de armazenagem, nos coeficientes de análise de trilha. As variáveis teor de umidade, conteúdo de cinzas, proteínas, lipídios, grãos sem defeitos, massa volumétrica, incidência de Penicillium sp., Aspergillus sp. e Fusarium sp. devem ser consideradas conjuntamente na definição da possibilidade de ocorrência de aflatoxinas na massa de grãos armazenados.

Characterization of rue extract and its potential for controlling rice blast

Abstract: The objective of this work was to purify and standardize the rue (Ruta graveolens) extract and evaluate its effect on Magnaporthe oryzae as an alternative to the integrated management of rice blast. The drug was characterized, the liquid extract was obtained, and the methodology for quantifying the standard markers psoralen and bergapten was validated. Rue extract and the markers, solely or in combination, were assayed in vitro, as well as in greenhouse conditions, for their ability to suppress leaf blast, by the evaluation of mycelial growth, conidial germination, and appressorium formation. Rue extract inhibited M. oryzae mycelial growth (100%), conidial germination (LD50=0.237 mg), and the appressorium formation (LD50=0.121 mg); besides, the extract reduced leaf blast severity by 80.84%. Fluorescence microscopy showed that rue extract did not damage M. oryzae cell wall and plasma membrane, indicating another mode of action. Rue extract has a great potential for controlling rice leaf blast.

Contributions of the peroxisome and β-oxidation cycle to biotin synthesis in fungi.

The first step in the synthesis of the bicyclic rings of D-biotin is mediated by 8-amino-7-oxononanoate (AON) synthase, which catalyzes the decarboxylative condensation of l-alanine and pimelate thioester. We found that the Aspergillus nidulans AON synthase, encoded by the bioF gene, is a peroxisomal enzyme with a type 1 peroxisomal targeting sequence (PTS1). Localization of AON to the peroxisome was essential for biotin synthesis because expression of a cytosolic AON variant or deletion of pexE, encoding the PTS1 receptor, rendered A. nidulans a biotin auxotroph. AON synthases with PTS1 are found throughout the fungal kingdom, in ascomycetes, basidiomycetes, and members of basal fungal lineages but not in representatives of the Saccharomyces species complex, including Saccharomyces cerevisiae. A. nidulans mutants defective in the peroxisomal acyl-CoA oxidase AoxA or the multifunctional protein FoxA showed a strong decrease in colonial growth rate in biotin-deficient medium, whereas partial growth recovery occurred with pimelic acid supplementation. These results indicate that pimeloyl-CoA is the in vivo substrate of AON synthase and that it is generated in the peroxisome via the β-oxidation cycle in A. nidulans and probably in a broad range of fungi. However, the β-oxidation cycle is not essential for biotin synthesis in S. cerevisiae or Escherichia coli. These results suggest that alternative pathways for synthesis of the pimelate intermediate exist in bacteria and eukaryotes and that Saccharomyces species use a pathway different from that used by the majority of fungi.

Preparation of (S)-1-halo-2-octanols using ionic liquids and biocatalysts

Preparation of (S)-1-chloro-2-octanol and (S)-1-bromo-2-octanol was carried out by the enzymatic hydrolysis of halohydrin palmitates using biocatalysts. Halohydrin palmitates were prepared by various methods from palmitic acid and 1,2-octanediol. A tandem hydrolysis was carried out using lipases from Candida antarctica (Novozym® 435), Rhizomucor miehei (Lipozyme IM), and “resting cells” from a Rhizopus oryzae strain that was not mycotoxigenic. The influence of the enzyme and the reaction medium on the selective hydrolysis of isomeric mixtures of halohydrin esters is described. Novozym® 435 allowed preparation of (S)-1-chloro-2-octanol and (S)-1-bromo-2-octanol after 1–3 h ofreaction at 40 °C in [BMIM][PF6].

Patulin in food: state-of-the-art and analytical trends

Patulin is a mycotoxin produced by several fungal species of the genera Penicillium and Aspergillus, found on several fruit species and, remarkably, in apples and apple products. Patulin has a broad spectrum of toxicity, including carcinogenicity and teratogenicity in animals. Due to the stability of the molecule, considerable amounts of patulin still remain in apple products after processing. This paper reviews different analytical methods for patulin determination and methods to reduce levels of patulin in apple products as well.