Production and partial characterization of polygalacturonases produced by thermophilic Monascus sp N8 and by thermotolerant Aspergillus sp N12 on solid-state fermentation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Stimulation of polygalacturonase production in an immobilized system by Aspergillus sp.: effect of pectin and glucose

The synthesis of polygalacturonases (PG) is known to be influenced by Aspergillus growth conditions, namely, environmental factors and pectin content in the cultivation medium containing a mixed carbon source. Optimal conditions were attained at a temperature of 30 A degrees C and an initial pH of 4.5. PG activity (3.29 and 2.48 U/mL) was determined after a two-day culture of Aspergillus sp. HC1 and Aspergillus sp. CC1, respectively, in a basic medium containing 2% citrus pectin as the sole carbon source. The addition of glucose (2% w/v) to the basic medium led to a 2-fold increase in PG production. However, enzyme synthesis was repressed when a higher concentration of glucose was used in the medium containing the mixed carbon source. Spores from the two fungi were immobilized in a 3% Ca-alginate system and the mechanical strength of the gel beads allowed the use of this process system 6-fold longer (288 h) than the free culture. In the Aspergillus sp. CC1 immobilized system, PG production increased nearly 10-fold in the medium with 2% glucose added (5.95 U/mL) in comparison to the medium without sugar (0.55 U/mL). The results demonstrate that a different response in activity was produced by free and entrapped spore systems. PG production remained approximately constant throughout the six 48 h cycles in the medium containing citrus pectin (2% w/v) as the sole carbon source.

Induction of physical paramorphogenesis in Aspergillus sp

A simple and easily reproducible method to obtain little fungical pellets from mycelial fractionization, completely under control with respect to development, size and homogeneity without genetic compromise, is described. The method has been successully employed with Aspergillus niger and also showed similar results with Aspergillus oryzae.

Biochemical characterisation and application of lipases produced by Aspergillus sp on solid-state fermentation using three substrates

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Produção e caracterização parcial da xilanase produzida por Aspergillus sp

As xilanases são enzimas que hidrolisam as ligações 1,4-β-xilosídicas da xilana e que possuem potencial biotecnológico em vários processos industriais, como na clarificação de sucos e vinhos, na fabricação de pães, na produção de biocombustíveis e no tratamento das polpas celulósicas. Além disso, os produtos de hidrólise da xilana, xilooligossacarídeos, podem ser utilizados como ingredientes prebióticos, ou seja, ingredientes nutricionais não digeríveis que estimulam seletivamente a proliferação e a atividade de bactérias benéficas do cólon. O uso de enzimas microbianas nas indústrias se dá devido às suas diversas vantagens sobre os métodos químicos e à facilidade de obtenção do microrganismo. Assim, os objetivos deste trabalho foram: produzir e caracterizar parcialmente a xilanase de Aspergillus sp isolado de pó de café utilizado, purificar parcialmente a enzima produzida e analisar os produtos de hidrólise da xilana. O Aspergillus sp mostrou-se bom produtor de xilanase quando o pó de sabugo de milho foi utilizado como fonte de carbono e a xilanase produzida apresentou melhor atividade específica quando o período de cultivo foi de 168 horas. Os processos de precipitação de proteínas e diálise promoveram o aumento da atividade específica do extrato. A xilanase de Aspergillus sp apresentou pH e temperatura de máxima atividade semelhantes aos de produzidas por Aspergillus niger isolados de outras fontes. A hidrólise da xilana produziu xilose e xilooligossacarídeos. Além da xilanase, o fungo revelou ser produtor de outras proteínas que podem ser estudadas em pesquisas futuras.

Diversidade de geofungos em águas do Parque Municipal do Ibirapuera na cidade de São Paulo, SP, Brasil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Desempenho de sementes de algodão tratadas quimicamente e armazenadas

Considerando que o armazenamento desempenha papel decisivo na manutenção da qualidade da semente, desenvolveu-se este trabalho com o objetivo de avaliar a qualidade fisiológica e fitossanitária de sementes de algodão, tratadas quimicamente e armazenadas por doze meses. Foram utilizados dois lotes de sementes de algodão cv. DeltaPine-AC90, deslintados quimicamente, que foram submetidos aos tratamentos fungicidas e inseticidas: testemunha; Disulfoton + Carboxin + Thiram; Carbofuran + Carboxin + Thiram; Imidacloprid + Tolylfluanid + Pencycuron. As sementes foram armazenadas em armazém sem controle de temperatura e umidade relativa do ar. Foram retiradas amostras de sementes no início do armazenamento e a cada dois meses e avaliadas quanto ao teor de água, à porcentagem de germinação, ao vigor (testes de envelhecimento acelerado e de germinação à baixa temperatura), à sanidade e à emergência das plântulas. O delineamento utilizado foi o inteiramente casualizado, em esquema fatorial, com quatro repetições. Verificou-se redução da germinação e do vigor de sementes em função do armazenamento. A redução da qualidade fisiológica associou-se com o aumento na ocorrência de Aspergillus sp. e Penicillium sp. nas sementes. A manutenção da qualidade de sementes de algodão foi obtida até o oitavo mês de armazenagem, podendo-se concluir que: a eficiência do tratamento químico de sementes de algodão depende da combinação de produtos utilizados; não se deve tratar com fungicida sementes de algodão com baixo nível de vigor; a ocorrência dos fungos Aspergillus sp. e Penicillium sp. aumentou com o período de armazenamento nas sementes sem tratamento e que a manutenção da qualidade de sementes de algodão para comercialização depende da sua qualidade inicial e do período de armazenamento.

Protease production by different thermophilic fungi

A comparative study was carried out to evaluate protease production in solid-state fermentation (SSF) and submerged fermentation (SmF) by nine different thermophilic fungi - Thermoascus aurantiacus Miehe, Thermomyces lanuginosus, T. lanuginosus TO.03, Aspergillus flavus 1.2, Aspergillus sp. 13.33, Aspergillus sp. 13.34, Aspergillus sp. 13.35, Rhizomucor pusillus 13.36 and Rhizomucor sp. 13.37 - using substrates containing proteins to induce enzyme secretion. Soybean extract (soybean milk), soybean flour, milk powder, rice, and wheat bran were tested. The most satisfactory results were obtained when using wheat bran in SSF. The fungi that stood out in SSF were T. lanuginosus, T. lanuginosus TO.03, Aspergillus sp. 13.34, Aspergillus sp. 13.35, and Rhizomucor sp. 13.37, and those in SmF were T. aurantiacus, T. lanuginosus TO.03, and 13.37. In both fermentation systems, A. flavus 1.2 and R. pusillus 13.36 presented the lowest levels of proteolytic activity.

Germinação, qualidade sanitária e armazenamento de sementes de canafístula (Peltophorum dubium)

Neste trabalho, objetivou-se avaliar o efeito de métodos de superação de dormência e do ambiente de armazenamento sobre a qualidade fisiológica e fitopatológica das sementes de canafístula (Peltophorum dubium). As sementes foram submetidas aos seguintes tratamentos de superação de dormência: escarificação com lixa (200); imersão em água na temperatura ambiente, durante 24 e 72 h; imersão em ácido sulfúrico por 2, 6, 10, 15, 20 e 30 min; imersão em água quente (70, 80 e 90 C); e umedecimento do substrato com solução de KNO3 (0,2%). As sementes foram armazenadas na temperatura ambiente e a 10 C por 210 dias. Os efeitos dos tratamentos e do armazenamento foram avaliados por meio do teor de água, teste de germinação (cinco repetições de 30 sementes), de comprimento de plântulas e sanidade (400 sementes), com incubação por oito dias (22-25 C). Na análise estatística dos dados, utilizou-se o delineamento experimental inteiramente casualizado em esquema fatorial 2 x 14 (condições de armazenamento x tratamentos para a superação da dormência). As médias foram comparadas pelo teste de Tukey (P>0,5). Com relação às sementes não armazenadas, os melhores tratamentos para superar a dormência e promover a germinação foram escarificação com lixa ou ácido sulfúrico por 15 a 30 min; quanto às sementes armazenadas, houve a imersão em água quente (70 a 80 ºC). Os fungos detectados nas sementes foram Pestalotia sp., Alternaria sp., Rhizopus sp., Nigrospora sp., Curvularia sp., Fusarium sp., Rhizoctonia sp., Aspergillus sp., Cladosporium sp. e Fusarium semitectum.

TRATAMENTO DE SEMENTES DE ARROZ, TRIGO, FEIJÃO E SOJA COM UM PRODUTO FORMULADO À BASE DE CÉLULAS E DE METABÓLITOS DE Bacillus subtilis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Influence of different drying temperatures on the amount of essential oils and citral content in Cymbopogon citratus (DC) Stapf. - Poaceae

Studies were conducted to show the effect of different temperatures in the drying process on the amount and quality of essential oils of Cymbopogon citratus (DC) Stapf. Leaves were harvested in the experimental field of the Agronomical Sciences College, UNESP, Botucatu, SP, Brazil in September, 1996. Blades of the leaves were cut in small parts (about 1-1,5 cm length), dried for several days at 30°, 50°, 70° and 90°C, until establishment of the weights. In the following process a hydrodistillation, during 2.5 hours, by Clevenger apparatus, was subsidized to extract the essential oils. A higher amount of oil could clearly be collected with the lower drying temperatures, except at 30°C, affected by fungus growing. Aspergillus sp., Penicillium sp., Rhyzopus sp., Cladosporium sp., Trichoderma sp. and Alternaria sp. were observed in the leaves. The analysis of the oil by GC-MS showed the variation of citral concentration of the treatments (86,1 to 95,2%). The results proved it is worthwhile to spend more time and effort in the production process using longer times of careful drying.

Isolamento de fungos produtores de enzimas pectinolíticas

Soil samples collected in the campus, UNESP, Araraquara, SP, were employed to isolate and characterize fungi strains with potential pectinolytic enzymes. These enzymes have arisen great interest due to its increasing application in the food industry. Two hundred forty six strains were isolated based on the appearance of colony on PDA medium, morphology (septate mycelia, nonseptate conidiophore, black conidia, and clublike spore-bearing head), after 48 h of growth at 30°C. Strains were selected in solid medium containing pectin citrus as sole carbon source and 0.5% rutenium red. The characterization of pectinolytic production was performed in solid culture and batch fermentation medium containing pectin citrus. The enzyme pectinolytic production was evaluated at 30°C, without agitation in 100 mL of medium containing 2% pectin citrus, 0.2% ammonium sulphate, 0.2% magnesium sulphate, and 0.05% potassium phosphate. The maximum pectinolytic activity (15U/mL) was observed in the medium after Aspergillus sp CFCF-0492 growth, while Aspergillus sp CFCF-CC1 showed the higher level of the final biomass. The pectinolytic activity is more preserved when the fungi-spores were maintained in agar-Czapeck medium.

Sementes de feijão-caupi (Vigna unguiculata L. (Walp), tratadas com extrato de folhas de nim (Azadirachta indica A. Juss.) avaliação da germinação e da incidência de fungos

This study aimed to evaluate the effect of aqueous extract of neem on germination and fungi incidence on seeds of three cultivars (Serrinha, BR 17 and Maranhão) of cowpea. Neem leaves were dryed, crushed and prepared dilutions of 0.5; 1.0; 2.0, 4.0 g dm -3and control. The fungi incidence was evaluated by the test filter paper and germination according to the Rules for Seeds Testing (Regras para Análise de Sementes). In the three cultivars analyzed, reduction in the incidence of Aspergillus sp and Fusarium sp was observed. In relation to the influence of extracts of neem leaves on seed germination, significant effect of extract in Maranhão cultivar was observed, where all concentrations differed from the control, and propovided a considerable increase in the percentage of normal seedlings. It was concluded that the leaf extract of neem was effective in controlling Aspergillus sp, Fusarium sp , Phoma sp and Macrophomina phaseolina at different concentrations in different cultivars and seed germination was stimulated for the Maranhão cultivar.

Estudo da mcrobiota fúngica gastrintestinal de morcegos (Mammalia, Chiroptera) da região noroeste do estado de São Paulo: Potencial zoonótico

Bats are hosts of a rich diversity of microorganisms. Many studies indicate a close link between bats and fungi with pathogenic potential, especially for living in environments such as caves, caverns and hollow trees, favorable to the maintenance and spread of fungi. The objective was to study the gastrointestinal mycoflora of bats. Of the 98 samples belonging to 11 species of bats coming from 15 studied cities, 20% of the species were Carollia perspicillata, 19% Artibeus lituratus, 17% Molossus rufus, 13% Glossophaga soricina, 9% Nyctinomops macrotis, 8% Molossus molossus, 7% Desmodus rotundus, 2% Lasiurus ega and 1% Eptesicus furinalis, Myotis nigricans and Tadarida brasiliensis. The genus Aspergillus sp. was isolated from 29% of the samples, followed by 6% Microsporum sp. and Penicillium sp. 4% Trichophyton sp. and zygomycetes and 2% Fusarium sp. Of yeast species, 14% were from Rhodotorula sp., 10% Candida sp. and 2% Cryptococcus sp., 22% of isolates remained unidentified. All 82 cultures of organs were negative for Histoplasma capsulatum. There was a statistically significant association between the results of microbiological culture and bat species (p < 0.05). We conclude that the bats can act as disperser agents of fungi with pathogenic potential, although other studies should be performed to establish strategies to identify the main factors correlated with the growth and spread of microorganisms in nature and implication of bats in the epidemiological cycle.

Identificação de espécies fúngicas nas onicomicoses do idoso institucionalizado

BACKGROUND: Superficial fungal infections are caused by dermatophytes, yeasts or filamentous fungi. They are correlated to the etiologic agent, the level of integrity of the host immune response, the site of the lesion and also the injured tissue. OBJECTIVE: The purpose of this study is to isolate and to identify onychomycosis agents in institutionalized elderly (60 years old +). METHODS: The identification of the fungi relied upon the com-bined results of mycological examination, culture isolation and micro cultures observation under light microscopy from nail and interdigital scales, which were collected from 35 elderly with a clinical suspicion of onychomycosis and a control group (9 elderly with healthy interdigital space and nails). Both groups were insti-tutionalized in two nursing homes in Sao Bernardo do Campo, SP, Brazil. RESULTS: The nail scrapings showed 51.40% positivity. Of these, dermatophytes were found in 44.40% isolates, 27.78% identified as Trichophyton rubrum and 5.56% each as Trichophyton tonsurans, Trichophyton mentagrophytes and Microsporum gypseum. The second more conspicuous group showed 38.89% yeasts: 16.67% Candida guilliermondii, 11.11% Candida parapsilosis, 5.56% Candida glabrata, and 5.56% Trichosporon asahii. A third group displayed 16.70% filamen-tous fungi, like Fusarium sp, Aspergillus sp and Neoscytalidium sp (5.56% each). The interdigital scrapings pre-sented a positivity rate of 14.29%. The agents were coincident with the fungi that caused the onychomycosis. In the control group, Candida guilliermondii was found at interdigital space in one person. CONCLUSION: Employing a combination of those identification methods, we found no difference between the etiology of the institutional-ized elderly onychomycosis from that reported in the literature for the general population. © 2013 by Anais Brasileiros de Dermatologia.