Inferência filogenética revela a complexa etiologia das manchas areolada e foliar em seringueira e em outras espécies cultivadas na Amazônia

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

Produção de enzimas por fungos em fermentação semi-sólida utilizando bagaço de coco e pedúnculo de caju como substratos

The production of enzymes by microorganisms using organic residues is important and it can be associated with several applications such as food and chemical industries and so on. The objective of this work is the production of CMCase, Xylanase, Avicelase and FPase enzymes by solid state fermentation (SSF) using as substrates: bagasse of coconut and dried cashew stem. The microorganisms employed are Penicillium chrysogenum and an isolated fungus from the coconut bark (Aspergillus fumigatus). Through the factorial design methodology and response surface analysis it was possible to study the influence of the humidity and pH. For Penicillium chrysogenum and the isolated fungus, the coconut bagasse was used as culture medium. In another fermentation, it was used the mixture of coconut bagasse and cashew stem. Fermentations were conducted using only the coconut bagasse as substrate in cultures with Penicillium chrysogenum fungus and the isolated one. A mixture with 50% of coconut and 50% of cashew stem was employed only for Penicillium chrysogenum fungus, the cultivation conditions were: 120 hours at 30 °C in BOD, changing humidity and pH values. In order to check the influence of the variables: humidity and pH, a 2 2 factorial experimental design was developed, and then two factors with two levels for each factor and three repetitions at the central point. The levels of the independent variables used in ascending order (-1, 0, +1), to humidity, 66%, 70.5% and 75% and pH 3, 5 and 7, respectively. The software STATISTICA TM (version 7.0, StatSoft, Inc.) was used to calculate the main effects of the variables and their interactions. The response surface methodology was used to optimize the conditions of the SSF. A chemical and a physic-chemical characterization of the coconut bagasse have determined the composition of cellulose (%) = 39.09; Hemicellulose (%) = 23.80, Total Lignin (%) = 36.22 and Pectin (%) = 1.64. To the characterization of cashew stem, the values were cellulose (g) = 15.91 Hemicellulose (%) = 16.77, Total Lignin (%) = 30.04 and Pectin (%) = 15.24. The results indicate the potential of the materials as substrate for semisolid fermentation enzyme production. The two microorganisms used are presented as good producers of cellulases. The results showed the potential of the fungus in the production of CMCase enzyme, with a maximum of 0.282 UI/mL and the Avicelase enzyme the maximum value ranged from 0.018 to 0.020 UI/ mL, using only coconut bagasse as substrate. The Penicillium chrysogenum fungus has showed the best results for CMCase = 0.294 UI/mL, FPase = 0.058 UI/mL, Avicelase = 0.010 UI/mL and Xylanase = 0.644 UI/ mL enzyme, using coconut bagasse and cashew stem as substrates. The Penicllium chrysogenum fungus showed enzymatic activities using only the coconut as substrate for CMCase = 0.233 UI/mL, FPase = 0.031 to 0.032 UI/ mL, Avicelase = 0.018 to 0.020 UI/mL and Xylanase = 0.735 UI/ mL. Thus, it can be concluded that the used organisms and substrates have offered potential for enzyme production processes in a semi-solid cultivation

Palma forrageira (Opuntia ficus indica e Nopalea cochenillifera) como matéria-prima para produção de etanol celulósico e enzimas celulolíticas

The need for new sources of energy and the concern about the environment have pushed the search for renewable energy sources such as ethanol. The use of lignocellulosic biomass as substrate appears as an important alternative because of the abundance of this raw material and for it does not compete with food production. However, the process still meets difficulties of implementation, including the cost for production of enzymes that degrade cellulose to fermentable sugars. The aim of this study was to evaluate the behavior of the species of cactus pear Opuntia ficus indica and Nopalea cochenillifera, commonly found in northeastern Brazil, as raw materials for the production of: 1) cellulosic ethanol by simultaneous saccharification and fermentation (SSF) process, using two different strains of Saccharomyces cerevisiae (PE-2 and LNF CA-11), and 2) cellulolytic enzymes by semi-solid state fermentation (SSSF) using the filamentous fungus Penicillium chrysogenum. Before alcoholic fermentation process, the material was conditioned and pretreated by three different strategies: alkaline hydrogen peroxide, alkaline using NaOH and acid using H2SO4 followed by alkaline delignification with NaOH. Analysis of composition, crystallinity and enzymatic digestibility were carried out with the material before and after pretreatment. In addition, scanning electron microscopy images were used to compare qualitatively the material and observe the effects of pretreatments. An experimental design 2² with triplicate at the central point was used to evaluate the influence of temperature (30, 40 and 45 °C) and the initial charge of substrate (3, 4 and 5% cellulose) in the SSF process using the material obtained through the best condition and testing both strains of S. cerevisiae, one of them flocculent (LNF CA-11). For cellulase production, the filamentous fungus P. chrysogenum was tested with N. cochenillifera in the raw condition (without pretreatment) and pretrated hydrothermically, varying the pH of the fermentative medium (3, 5 and 7). The characterization of cactus pear resulted in 31.55% cellulose, 17.12% hemicellulose and 10.25% lignin for N. cochenillifera and 34.86% cellulose, 19.97% hemicellulose and 15.72% lignin for O. ficus indica. It has also been determined, to N. cochenillifera and O. ficus indica, the content of pectin (5.44% and 5.55% of calcium pectate, respectively), extractives (26.90% and 9.69%, respectively) and ashes (5.40% and 5.95%). Pretreatment using alkaline hydrogen peroxide resulted in the best cellulose recovery results (86.16% for N. cochenillifera and 93.59% for O. ficus indica) and delignification (48.79% and 23.84% for N. cochenillifera and O. ficus indica, respectively). This pretreatment was also the only one which did not increase the crystallinity index of the samples, in the case of O. ficus indica. However, when analyzing the enzymatic digestibility of cellulose, alkali pretreatment was the one which showed the best yields and therefore it was chosen for the tests in SSF. The experiments showed higher yield of conversion of cellulose to ethanol by PE-2 strain using the pretreated N. cochenillifera (93.81%) at 40 °C using 4% initial charge of cellulose. N. cochenillifera gave better yields than O. ficus indica and PE-2 strain showed better performance than CA-11. N. cochenillifera proved to be a substrate that can be used in the SSSF for enzymes production, reaching values of 1.00 U/g of CMCase and 0.85 FPU/g. The pretreatment was not effective to increase the enzymatic activity values

Strain differentiation of Trichophyton rubrum by random amplification of polymorphic DNA (RAPD)

Trichophyton rubrum é um importante agente causal de dermatomicose. Os métodos de tipagem molecular têm sido recentemente desenvolvidos para responder questões sobre epidemiologia e auxiliar no esclarecimento de recidivas, após o tratamento. As seqüências aleatórias 1- (5'-d[GGTGCGGGAA]-3') e 6- (5'-d[CCCGTCAGCA]-3') foram usadas para tipagem molecular deste fungo por RAPD produzindo variabilidade intraespecífica. Cinco padrões foram observados entre os 10 isolados de T. rubrum, com ambas as seqüências. Foi concluído que a análise por RAPD pode ser utilizada para estudos epidemiológicos.

Atividade antifúngica de extratos de Momordica charantia L. sobre Colletotrichum musae

Os objetivos do presente trabalho foram avaliar os efeitos de extratos de Momordica charantia sobre o crescimento micelial e a germinação de conídios de Colletotrichum musae, e a eficiência destes extratos no controle da antracnose, causada por C. musae, em bananas. Extratos aquoso e hidroetanólico, obtidos de folhas e ramos, na concentração de 50% em relação ao volume adicionado, em meio sólido, proporcionaram 71 e 65% de inibição do crescimento micelial, respectivamente, enquanto que em meio líquido, a inibição do crescimento micelial foi de 86 e 81%, respectivamente. Somente o extrato aquoso e o tiofanato metílico, nas concentrações de 50% e 1000 µg mL-1 respectivamente, proporcionaram 100% de inibição da germinação de esporos de C. musae. Os extratos metanólico e aquoso inibiram em 80 e 70%, respectivamente, o desenvolvimento das lesões em bananas, quando aplicados até dois dias antes da inoculação do fungo. Estes resultados foram semelhantes ao tratamento com tiofanato metílico, que inibiu 80% do desenvolvimento das lesões. Confirma-se a presença de substância antifúngica nos extratos de M. charantia e outros estudos devem ser realizados para viabilizar seu uso no controle da antracnose da banana.

The hamster cheek pouch: an immunologically privileged site suitable to the study of granulomatous infections

A bolsa jugal do hamster (BJH) é uma invaginação da mucosa oral, caracterizada histologicamente como semelhante a pele. Nesse estudo nós descrevemos algumas de suas características anatômicas, histológicas e embriológicas e comentamos sobre sua propriedade como local imunologicamente privilegiado, considerando a ausência de drenagem linfática e o reduzido número de células de Langerhans. Apresentamos também os resultados obtidos quando da inoculação de micobacterias (BCG, Mycobacterium tuberculosis e Mycobacterium leprae) e do fungo Paracoccidioides brasiliensis na bolsa jugal. Comparada com as lesões provocadas em outras localizações e, à exceção do BCG, as lesões induzidas na bolsa são menores e de maior duração e, mesmo quando granulomatosas, incapazes de controlar a multiplicação do agente; nos casos em que houve o desenvolvimento da resposta imune, ele se fez tardiamente e foi acompanhado pela redução do número de parasitas nas lesões. Essas observações apontam a bolsa jugal do hamster como um local de escolha para o estudo sobre a participação da resposta imune no desenvolvimento e modulação das doenças infecciosas e dos granulomas.

Experimental dermatophytosis in hamsters inoculated with Trichophyton mentagrophytes in the cheek pouch

Esse estudo apresenta os resultados obtidos quando da inoculação de Trychophyton mentagrophytes na bolsa jugal do hamster, local imunologicamente privilegiado. Foram utilizados 42 animais: 21 inoculados com 10(6) fungos na bolsa jugal (grupo 1) e, 21 inicialmente inoculados com 10(6) fungos no coxim plantar e, 15 dias após, na bolsa jugal com a mesma quantidade fúngica (grupo 2). Os animais foram sacrificados às 20 h, 3, 7, 14, 30, 60 e 120 dias; foram coletadas amostras da bolsa jugal inoculada, e das patas submetidas ao teste do coxim plantar (TCP). Independente do grupo e do tempo de evolução da infecção, os animais não desenvolveram hipersensibilidade tardia avaliada através do TCP. A pré-inoculação de fungos no coxim plantar não alterou a morfologia das lesões induzidas na bolsa jugal. Assim, nos animais do grupo 1 e grupo 2, a introdução do fungo na bolsa jugal, resultou em lesão focal, constituída por infiltrado inflamatório agudo estéril, com formação de abscesso, que evoluiu para reação macrofágica e, posteriormente, para a resolução mesmo na ausência de resposta imune detectável pelo TCP. Nossos resultados indicam que, apesar do importante papel da resposta imune na regressão espontânea da dermatofitose, outros fatores são, também, parte integral da defesa contra esta infecção fúngica.

Dermatophytosis: association between ABO blood groups and reactivity to the trichophytin

Os autores investigaram a relação entre dermatofitose e grupo sanguíneo ABO através da tipagem sanguínea, identificação do dermatófito isolado e resposta imune celular específica de 40 indivíduos portadores desta micose. Verificaram que o fungo Trichophyton rubrum foi isolado em 54,5% dos pacientes, sendo mais frequente em indivíduos pertencentes ao grupo sanguíneo A. A resposta imune celular, avaliada através do antígeno tricofitina, foi positiva em 25% dos pacientes estudados; a presença de reações imediatas (30 min) foi verificada em 35%. A distribuição dos grupos sanguíneos entre pacientes com dermatofitose e grupo controle foi a seguinte: 47,5% x 36% grupo A, 40% x 50% grupo O, 12,5% x 11% grupo B. Embora os autores tenham encontrado um número maior de pacientes pertencentes ao grupo sanguíneo A e infectados pelo T. rubrum, não obtiveram evidência estatística de que esses indivíduos sejam mais suscetíveis as dermatofitoses.

Estrutura genética de populações de Amphobotrys ricini, agente causal do mofo cinzento da mamoneira

The gray mold, causal organism Amphobotrys ricini, is one of the major diseases of castor bean. Difficulties in managing plant disease arises form the limited understanding of the genetic structure of A. ricini, their complexity and variability make it difficult to control. Genetic structure can be used to infer the relative impact of different forces that influence the evolution of pathogen populations, that allow to predict the potencial for pathogen populations to envolve in agricultural ecosystems. Growers protect their crop by applying fungicides, but there aren t fungicides to provide significant control of gray mold of castor bean. The objectives of this work were use RAPD to determine the genetic structure of A. ricini subpopulations in Paraíba and assay the sensitivity of A. ricini isolates to azoxystrobin and carbendazim. To determine the genetic structure of A. ricini subpopulations in Paraíba, 23 isolates were colleted from two different geographic location (subpopulation). These isolates were analysed by RAPD using 22 random decamer primers, purchased from OPERON, produced a total of 80 markers polimorphics. The resulting matrixes were analysed using PopGene version 1.32. Sensitivity to azoxystrobin and carbendazim of 30 isolates, colleted form Paraíba and Alagoas, was estimated based on spore germination and colony growth inhibition. The stock solutions were added toV8 medium after sterilization to produce final concentrations of 0, 0.01, 0.1, 1, 10, and 100 µg/ml of carbendazim and 0, 0.001, 0.01, 0.1, 1, and 10 µg/ml of azoxystrobin. All statistical analyses were performed using SAS to estimate the dose that inhibited fungal growth by 50% (ED50 values). The genetic diversity within subpopulations (Hs=0,271) accounted for 92% of the total genetic diversity (Ht=0,293), while genetic diversity between subpopulations (Gst = 0,075) represented only 7,5%. The estimated number of migrants per generation (NM ) was 6,15. Nei s average gene identity across 80 RAPD loci was 0,9468. Individual ED50 values, for the 30 isolates screened for their sensitivity to azoxystrobin, ranged From a maximum of 0,168 µg/ml to a minimum of 0,0036 µg/ml. The ED50 values for carbendazim varied within the range of 0,026 to 0,316 µg/ml

Estudo etnofarmacológico e fitoquímico das espécies medicinais Cleome spinosa Jacq, Pavonia varians Moric e Croton cajucara Benth

In this present work an ethnographic research was performed with 84 native medicinal specimens from the Litoral Norte Riograndense, from which two plants Cleome spinosa Jacq e Pavonia varians Moric were submitted to ethnobotanic, phytochemistry and pharmacologic investigations. Additionally, a phytopharmacological research of the medicinal specimen Croton cajucara Benth ( native plant of the Amazon region of Brazil) was improved. The obtained phytochemical results of the C. spinosa and P. varians showed the presence of flavonoids constituents, among other components. The two flavonoids (2S)-5-hydroxy-7,4 -dimethoxy-flavanone and 5,4 -dihydroxy-3,7,3 -trimethoxy-flavone were isolated from C. spinosa. The antioxidant activity of the hydroalcoholic extracts of C. spinosa and P. varians solubilized in the microemulsion systems SME-1 and SME-4, was evaluated in the DPPHmethod. The used SME systems [obtained with Tween 80: Span 20 (3:1) and isopropyl myristate (IPM)] improved the dissolution of those tested polar extracts, with higher efficacy to the SME-1 system (in which ethanol was included as cosurfactant). The CE50 values evidenced for P. varians were 114 [g/mL (SME-1) and 246 [g/mL (SME-4); for C. spinosa it was 224 [g/mL (SME-1) and 248 [g/mL (SME-4), being the system SME-1 more effective for both tested extracts. The hydroalcoholic extracts of P. varians (HAE-PV) was also submitted to pharmacological screening for antinociceptive activity in animal models. The oral administration of this extract (100, 300 and 1000 mg/kg) inhibited the acetic acid-induced writhing in mice. The higher inhibition (74%) was evidenced to the 1000 mg/kg administered dose. Its effect on the central nervous system (CNS) was investigated by tail flick and formalin-method and reveled that it has negligible antinociceptive action on the CNS. After taking consideration of HAE-PV interaction, Pavonia varians Moric could be used as a potent analgesic agent in case of peripheral algesia, without affecting the CNS. The phytochemical study of the stem bark of Croton cajucara Benth lead to the isolation of 19-nor-clerodanetype diterpenes, as well as to the separation of its fixed oil FO-CC. This non polar oil material reveled to be rich in sesquiterpenes and 19-nor-clerodanes components. The biologic effect of OF-CC was evaluated in the development in vitro of the fungis phytopatogens such as Fusarium oxysporum, Rhizoctonia solani and Sclerotium rolfsii. Significant inhibitory effect of the tested fungis (at 0,2 mg.mL-1 dosage) were comproved. A Mass Spectrometry study of clerodane-type diterpenes was developed in order to identify characteristic fragments on mass spectrometra of both clerodane and 19-nor-clerodane presenting an α,β-insaturated carbonyl moiety at ring A of the decalin-system. For that study, mass spectroscopy data were analysed for 19-nor-clerodanes [trans-dehydrocrotonin (DCTN), trans-crotonin (CTN), cis-cajucarin B (c-CJC-B), and cajucarinolide (CJCR)] and for clerodanes [isosacacarin (ISCR) and transcajucarin A (t-CJC-A)] obtained from the stem bark of C. cajucara, and also clerodane-type from other species. The trans-junction of the enone-system clerodanes was clear correlated with the presence of the characteristic ions at m/z 95, 121 e 205. Meanwhile, the characteristics ions at m/z 122 e 124 were correlated to cis-junction. The trans-junction of the enone-system 19-nor-clerodanes showed characteristics ions at m/z 161, 134 e 121. This study could be successful employed for identification of clerodane constituents from other specimens without any additional spectroscopic analyses, as well as a previously phytochemical analyzes in clerodane project search

Patogenicidade cruzada de Ceratobasidium spp. do caquizeiro (Diospyros kaki) e do chá(Camellia sinensis) e reação de cultivares de caqui ao patógeno

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

Atividade fungitóxica de Momordica charantia L. no controle de Sclerotium rolfsii Sacc.

O fungo Sclerotium rolfsii causa grandes perdas em algumas culturas econômicas. Por produzir estruturas de resistência (escleródios), este fungo é de difícil controle. Há escassez de novos ingredientes ativos eficientes para o controle deste patógeno. Assim, o objetivo do presente trabalho foi verificar se existe atividade fungitóxica na planta Momordica charantia (melão-de-sãocaetano), com potencial futuro para ser estudado no controle de S. rolfsii. Para isso, dois ensaios foram realizados, um in vitro (laboratório) e outro in vivo (câmara de crescimento). em in vitro, escleródios do patógeno ficaram em contato com extratos hidroetanólico e aquoso de folhas e ramos de M. charantia e sem extrato por 7, 14, 21 e 28 dias. A sobrevivência dos escleródios foi avaliada em meio de cultura específico, após cada tempo. em in vivo, testou-se a ação dos mesmos extratos de maneira preventiva e curativa (aplicação aos 6 e 3 dias antes do plantio; no dia do plantio; e aos 3 e 6 dias após o plantio) e no tratamento de semente, no patossistema feijoeiro cv. Carioquinha versus S. rolfsii. A eficiência da ação dos extratos foi avaliada por meio da severidade da doença. Os extratos hidroetanólico e aquoso, in vitro, de forma semelhante, controlaram 100% os escleródios, num período de 0 a 7 dias. No ensaio in vivo, o extrato hidroetanólico, aplicado tanto em 6 ou 3 dias, antes do plantio, de forma preventiva, diminuiu a severidade da doença em 74%. Há atividade fungitóxica na parte aérea da planta de melão-de-são-caetano, com potencial futuro de estudo para controlar S. rolfsii, preferencialmente, de maneira preventiva.

Severidade da mela da soja causada por Rhizoctonia solani AG-1 IA em função de doses de potássio

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

Imunoterapia de úlceras venosas com ß-(1-3) glucana insolúvel

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Pythium insidiosum: relato do primeiro caso de infecção humana no Brasil

A pitiose é causada por microorganismo aquático, fungo-símile, o Pythium insidiosum, patógeno de homens e animais. Observou-se um paciente com úlcera fagedênica no membro inferior, com exame anatomopatológico sugestivo de zigomicose, pouco sensível à terapêutica antifúngica, obtendo-se cura por meio de ampla exérese. A comprovação etiológica resultou de métodos moleculares, com amplificação e seqüenciamento de DNA de organismo isolado em ágar Sabouraud, observando-se 100% de analogia com seqüências de P. insidiosum depositadas no GenBank.