Seleção de isolados de fungos entomopatogênicos para o controle de Leptopharsa heveae (Hemiptera: Heteroptera, Tingidae)

Foram analisados vinte e sete isolados de fungos entomopatogênicos procedentes de diversos hospedeiros e regiões. A produção de conídios dos patógenos foi efetuada em placas de Petri contendo meio de cultura BDA. Insetos de terceiro instar de Leptopharsa heveae foram inoculados com uma suspensão fúngica na concentração de 1 x 10(9) conídios/mL. Avaliações de mortalidade confirmada foram efetuadas a cada dois dias e evidenciaram grande variação de virulência, sendo o isolado 1.189, de Metarhizium anisopliae, o mais virulento. Os cinco melhores isolados, mais os dois tidos como padrão por empresas do setor de heveicultura, foram então submetidos aos testes de produção. Para tanto, pré-matrizes em placas de Petri contendo meio de cultura BDA foram preparadas. A partir destas placas, matrizes contendo arroz como meio de cultura foram inoculadas com suspensão de conídios originária das pré-matrizes na concentração de 5 x 10(7) conídios/mL. Concluído o desenvolvimento dos patógenos nas matrizes, sacos de polipropileno contendo canjica de milho como meio de cultura foram inoculados com 40 mL da suspensão obtida a partir das matrizes. Após 15 dias de incubação, o conteúdo das sacolas foi avaliado quanto à produção e viabilidade de conídios. O isolado 1200, de Isaria fumosorosea, apresentou os melhores resultados de rendimento total e taxa de viabilidade e o isolado E9 de Metarhizium anisopliae foi o mais virulento ao percevejo de renda.

Potential of non-pathogenic Fusarium oxysporum isolates for control of Fusarium wilt of tomato

O trabalho avaliou a eficiência dos isolados (141/3, 233, 233/1, 245, 245/1, 251, 251/2, 251/5 e 257) de Fusarium oxysporum não patogênico ao tomateiro (Lycopersicon esculentum), no controle da murcha vascular causada por Fusarium oxysporum f. sp. lycopersici, raça 2 em plântulas de tomateiro cv. Viradoro. Para verificar o efeito dos isolados de F. oxysporum não patogênicos, o sistema radicular de plântulas de tomateiro, com 30 dias de idade, foi imerso na suspensão de conídios (10(6) ml-1) e as mudas transplantadas para substrato de cultivo. Após 35 dias do transplante foi verificado que esses isolados não foram patogênicos às plantas de tomateiro, nem afetaram o desenvolvimento das mudas. A eficiência dos isolados de Fusarium oxysporum não patogênicos no controle da murcha foi determinada imergindo-se as raízes de mudas de tomateiro em suspensão de conídios (10(6) conídios ml-1) e transplantando-as em substratos previamente infestados com os isolados de F. oxysporum f.sp. lycopersici, raça 2 (10(5) conídios ml-1 de substrato). Transcorridos 35 dias do transplante, foram realizadas as avaliações da severidade na escala de 1=planta sadia a 6=planta morta ou com vasos coloridos e folhas murchas até o ponteiro e altura das mudas. Os isolados de F. oxysporum não patogênicos foram eficientes em reduzir a severidade da doença e em manter normal o seu desenvolvimento. Esses resultados evidenciam a atividade antagônica dos isolados de F. oxysporum não patogênico no controle da murcha vascular do tomateiro, causada por Fusarium oxysporum f. sp. lycopersici raça 2.

Qualidade sanitária e germinação de sementes de pinhão-manso

O pinhão-manso é uma espécie arbustiva que está se destacando pelo interesse para produção de biocombustível, porém, os estudos sobre controle de qualidade das sementes estão se iniciando. Neste trabalho objetivou-se estudar o efeito da temperatura e do tempo de contagem na germinação de sementes de pinhão-manso, conduzido em papel toalha bem como avaliar a qualidade sanitária das sementes, desinfestadas ou não com hipoclorito de sódio, a 2%. O estudo foi conduzido, com sementes de pinhão-manso extraídas de frutos maduros, coletados na planta. O teste de germinação foi conduzido nas temperaturas de 25 ºC e alternadas de 20-30 ºC, com contagens aos 6, 9 e 12 dias. O teste de sanidade foi conduzido a 20 ºC com fotoperíodo de 12 h por 7 dias. Foi utilizado o delineamento experimental inteiramente casualizado com quatro repetições e empregado o teste F na análise de variância dos dados de germinação e do índice de velocidade de germinação. As médias foram comparadas pelo teste de Tukey a 5% de probabilidade. O potencial máximo de germinação das sementes de pinhão-manso foi obtido na temperatura de 25 ºC, com contagens aos seis, nove e 12 dias, entretanto não apresentou diferença significativa com as temperaturas de 20-30 ºC, nas contagens aos 9 e 12 dias. As sementes de pinhão-manso analisadas apresentaram alta frequência de espécies fúngicas, independente da desinfestação superficial. A incidência de fungos em ordem decrescente foi dos gêneros: Alternaria, Macrophomina, Cladosporium, Colletotrichum, Fusarium, Pestalotiopsis, Phoma, Helminthosporium, Epicocum e Nigrospora a maioria destes fungos apresenta potencial patogênico.

Influence of culture media in viability test of conidia of entomopathogenic fungi

O presente trabalho objetivou investigar se meios de cultura utilizados em teste de viabilidade afetam a germinação de conídios de cinco isolados de Lecanicillium lecanii, cinco de Beauveria bassiana e quatro de Paecilomyces fumosoroseus. Os testes foram realizados em lâminas de microscopia contendo um dos seguintes meios de cultura: Ágar-água (AA), Meio Mínimo (MM), Batata, dextrose e ágar (BDA), Batata, dextrose, ágar e 1% de extrato de levedura (BDAL), Sabouraud, dextrose, ágar e extrato de levedura (SDAL) e Meio Completo (MC). Delimitaram-se três áreas por lâmina e em cada uma aplicou-se 0,05mL de uma suspensão com concentração de 5,5 x 105 conídios ml-1. Para cada isolado foi realizado um bioensaio, com seis tratamentos e cinco repetições. Avaliou-se a germinação 15 horas após incubação, a 26±0,5ºC. Os meios de cultura influíram na capacidade de germinação das três espécies estudadas, ocorrendo variações inter e intraespecíficas. Verificou-se que os meios Completo e BDA proporcionaram as maiores porcentagens de germinação dos isolados de L. lecanii, sendo que e as menores foram obtidas nos meios SDAL e AA. Os meios ricos em nutrientes (BDA, BDAL e Completo) favoreceram a germinação dos isolados de B. bassiana, o que não ocorreu com os meios pobres (AA e MM). Nos meios Completo e BDA foram obtidas as maiores porcentagens de germinação dos isolados de P. fumosoroseus. As menores percentagens, por sua vez, foram obtidas no meio SDAL. Entretanto, alguns isolados apresentaram alta germinação em meios pobres em nutrientes (AA e MM).

Pathogenic variation in Colletotrichum gloeosporioides infecting Stylosanthes spp. in a center of diversity in Brazil

Pathogenic variation in Colletotrichum gloeosporioides infecting species of the tropical pasture legume Stylosanthes at its center of diversity was determined from 296 isolates collected from wild host population and selected germ plasm of S. capitata, S. guianensis, S. scabra, and S. macrocephala in Brazil. A putative host differential set comprising 11 accessions was selected from a bioassay of 18 isolates on 19 host accessions using principal component analysis. A similar analysis of anthracnose severity data for a subset of 195 isolates on the 11 differentials indicated that an adequate summary of pathogenic variation could be obtained using only five of these differentials. of the five differentials, S. seabrana 'Primar' was resistant and S. scabra 'Fitzroy' was susceptible to most isolates. A cluster analysis was used to determine eight natural race clusters using the 195 isolates. Linear discriminant functions were developed for eight race clusters using the 195 isolates as the training data set, and these were applied to classify a test data set of the remaining 101 isolates. All except 11 isolates of the test data set were classified into one of the eight race clusters. Over 10% of the 296 isolates were weakly pathogenic to all five differentials and another 40% were virulent on just one differential. The unclassified isolates represent six new races with unique virulence combinations, of which one isolate is virulent on all five differentials. The majority of isolates came from six field sites, and Shannon's index of diversity indicated considerable variation between sites. Pathogenic diversity was extensive at three sites where selected germ plasm were under evaluation, and complex race clusters and unclassified isolates representing new races were more prevalent at these sites compared with sites containing wild Stylosanthes populations.

Xylanases from fungi: properties and industrial applications

Xylan is the principal type of hemicellulose. It is a linear polymer of beta-D-xylopyranosyl units linked by (1-4) glycosidic bonds. In nature, the polysaccharide backbone may be added to 4-O-methyl-alpha-D-glucuronopyranosyl units, acetyl groups, alpha-L-arabinofuranosyl, etc., in variable proportions. An enzymatic complex is responsible for the hydrolysis of xylan, but the main enzymes involved are endo-1,4-beta-xylanase and beta-xylosidase. These enzymes are produced by fungi, bacteria, yeast, marine algae, protozoans, snails, crustaceans, insect, seeds, etc., but the principal commercial source is filamentous fungi. Recently, there has been much industrial interest in xylan and its hydrolytic enzymatic complex, as a supplement in animal feed, for the manufacture of bread, food and drinks, textiles, bleaching of cellulose pulp, ethanol and xylitol production. This review describes some properties of xylan and its metabolism, as well as the biochemical properties of xylanases and their commercial applications.

Antigenic similarities to Paracoccidioides brasiliensis in thermo-dependent dimorphic fungi isolated from soil in Botucatu, SP, Brazil

We compared the antigenic characteristics of two thermo-dependent dimorphic fungi isolated from soil in Botucatu, an endemic area of paracoccidioidomycosis (PCM) and Paracoccidioides brasiliensis. The soil isolates grew as cerebriform colonies at 37 degrees C (yeast form) and as cottonous colonies at 25 degrees C (mycelial form). No pathogenicity for ddY mice or hamsters were observed. In immunodiffusion test, there were precipitation bands between the 2 soil isolates and pooled PCM patient sera. There were also common precipitation bands at 21, 50 and 58 kDa between the soil isolates antigens and PCM patient sera by Western-blotting, but no gp43 kDa band. No gene for gp43 kDa protein was detected in the soil isolates by PCR. The fact that these isolates were obtained from an endemic area of PCM and there were some antigenic similarities between the soil isolates and P. brasiliensis in immunodiffusion test and Western-blotting may have some importance in epidemiological surveys done with paracoccidioidin as well interfering with the immune response of the exposed population.

EVALUATION OF DIFFERENT TECHNIQUES FOR THE DIFFERENTIATION OF PATHOGENIC AND NON PATHOGENIC STRAINS OF YERSINIA-ENTEROCOLITICA

Different methods and tests have been used to evaluate the pathogenic potential of distinct Y. enterocolitica serotypes and biotypes. We tested a total of 60 Y. enterocolitica strains, being 25 of human origin (serotype O3 biotype 4 and serotype O5 biotype 1); 6 of animal origin (serotype O3 biotype 4); 19 isolated from the environment (serotype O5.27 biotypes 1 and 2); and 8 isolated from food (serotype O5 biotype 1 and serotype 05.27 biotype 1). The methods used were based on plasmid gene expression (autoagglutination, calcium-dependence at 37 degrees C and Congo Red absorption tests), chromosomal gene expression (assays for pyrazinamidase activity, salicin fermentation and esculin hydrolysis), and invasion of HEp-2 cells. All but one of the Y. enterocolitica O3 strains, were found to be potentially pathogenic when submitted to the pyrazinamidase-salicin-esculin tests. In contrast, the results obtained with the assays related to plasmidial gene expression were not so uniform, probably due to plasmid loss. The least homogeneous results were obtained with the HEp-2 cell invasion test. Y. enterocolitica O5 behaved in a uniform manner when tested with the first two groups of tests (based on chromosomal and plasmidial gene expression), but not when tested with the HEp-2 invasion assay. The strains of serotype O5.27 biotype 1 presented a uniform behavior hen submitted to the chromosomic-related tests, showing no pathogenicity. However, they did not provide conclusive results with the tests related to plasmidial gene expression or HEp-2 cell invasion. We conclude that the tests related to chromosomal gene expression (esculin-salicin-pyrazinamidase) are simple and highly effective for the detection of potentially pathogenic Y. enterocolitica isolated from clinical cases.

Variability of non-mutualistic filamentous fungi associated with Atta sexdens rubropilosa nests

A survey of the filamentous fungi other than the symbiotic one found in association with Atta sexdens rubropilosa colonies was carried out. Different fungal species (27 taxa) were isolated a few days after treating the workers with toxic baits (sulfluramid; Mirex-SO), from 40 laboratory and 20 field nests. Syncephalastrum racemosum (54 %) and Escovopsis weberi (21 %), Trichoderma harzianum (38 %) and Fusarium oxysporum (23 %) were the prevalent species in laboratory and field nests, respectively. Acremonium kiliense, Acremonium strictum, E. weberi, F oxisporum, Fusarium solani, Moniliella silaveolens and T harzianum were found in both nests' groups. We revealed that many filamentous fungi can co-exist in a dormant state inside the nests of these insects and some of them appear to be tightly associated with this environment.

Decolorization of textile indigo dye by ligninolytic fungi

The indigo dye is extensively used by textile industries and is considered a recalcitrant substance, which causes environmental concern. Chemical products used on textile processing, which affect the environment through effluents, can be voluminous, colored and varied. Vat textile dyes, like indigo, are often used and dye mainly cellulosic fibers of cotton. Decolorization of this dye in liquid medium was tested with ligninolytic basidiomycete fungi from Brazil. Decolorization started in a few hours and after 4 days the removal of dye by Phellinus gilvus culture was in 100%, by Pleurotus sajor-caju 94%, by Pycnoporus sanguineus 91% and by Phanerochaete chrysosporium 75%. No color decrease was observed in a sterile control. Thin layer chromatography of fungi culture extracts revealed only one unknown metabolite of Rf = 0.60, as a result of dye degradation. (C) 2001 Published by Elsevier B.V. B.V.

Pathogenicity of isolates of Metarhizium anisopliae (Metsch.) Sorokin towards the cattle tick Boophilus microplus (Can.) (Acari : Ixodidae) under laboratory conditions

The pathogenic activity of Metarhizium anisopliae on Boophilus microplus females was assessed using five fungal isolates. Groups of 8 engorged ticks collected from cattle on free pasture were inoculated by immersion for 5 seconds in a spore suspension containing 2.5 x 10(8) conidia/ml. The groups were transferred to moist chambers in which they were maintained at 27 +/- 1 degreesC for about 15 days, their mortality being determined every 2, or occasionally 3, days. The isolate that produced the best results in this assay was used to inoculate female ticks with spore suspensions containing 0, 7.5 x 105, 7.5 x 10(6), 7.5 x 10(7) and 7.5 x 10(8) conidia/ml, under the same conditions as described above. The pathogenic action of M. anisopliae on engorged females was clearly demonstrated. Isolates E9 and AM were more effective, causing high tick mortality as well as reduced oviposition. The concentration of 7.5 x 10(8) conidia/ml was the most effective, with the fungus sporulating on 91.1% of the ticks. Mean percent oviposition was highest in the control treatment and lowest in the treatment with 7.5 x 10(8) conidia/ml.