The homeostasis of iron, copper, and zinc in paracoccidioides brasiliensis, cryptococcus neoformans var. Grubii, and cryptococcus gattii: a comparative analysis

Iron, copper, and zinc are essential for all living organisms. Moreover, the homeostasis of these metals is vital to microorganisms during pathogenic interactions with a host. Most pathogens have developed specific mechanisms for the uptake of micronutrients from their hosts in order to counteract the low availability of essential ions in infected tissues. We report here an analysis of genes potentially involved in iron, copper, and zinc uptake and homeostasis in the fungal pathogens Paracoccidioides brasiliensis, Cryptococcus neoformans var. grubii, and Cryptococcus gattii. Although prior studies have identified certain aspects of metal regulation in Cryptococcus species, little is known regarding the regulation of these elements in P. brasiliensis. We also present amino acid sequences analyses of deduced proteins in order to examine possible conserved domains. The genomic data reveals, for the first time, genes associated to iron, copper, and zinc assimilation and homeostasis in P. brasiliensis. Furthermore, analyses of the three fungal species identified homologs to genes associated with high-affinity uptake systems, vacuolar and mitochondrial iron storage, copper uptake and reduction, and zinc assimilation. However, homologs to genes involved in siderophore production were only found in P. brasiliensis. Interestingly, in silico analysis of the genomes of P. brasiliensis Pb01, Pb03, and Pb18 revealed significant differences in the presence and/or number of genes involved in metal homeostasis, such as in genes related to iron reduction and oxidation. The broad analyses of the genomes of P. brasiliensis, C. neoformans var. grubii, and C. gattii for genes involved in metal homeostasis provide important groundwork for numerous interesting future areas of investigation that are required in order to validate and explore the function of the identified genes and gene pathways.

Efeito da modificação atmosférica em goiabas var. Paluma na redução de danos mecânicos em pós-colheita

In the postharvest management, the fruits can be exposed to injuries that depreciated the quality and the shelf life. Thus, it was evaluated the modified atmosphere effects on guavas var. Paluma subjected to different mechanical damages. Once harvested, the fruits were selected, sanitized and submitted to the treatments T1 (control) - without injuries or packaging in bags of low density polyethylene (LDPE); T2 - without injuries + LDPE bags; T3 - damage by fall of 1 m + LDPE bags; T4 - damage by compression of 9 N + LDPE bags; T5 - damage by fall of 1 m + damage by compression of 9 N + LDPE bags and T6 - damage by fall of 1 m + damage by compression of 9 N without LDPE bags. The treatments were kept in cold storage at 10 ± 1 o C and 94 ± 2% de R.H. The analysis of CO2/ethylene production, enzymatic activity, total and soluble pectins, pulp firmness, titratable acidity (TA), soluble solids (SS), reducing sugars and ascorbic acid were performed every 10 days of refrigeration, and an additional day outside cold storage (22 ± 1o C and 75 ± 3% R.H.) for 30 days. Guavas packed in LDPE bags, not subject to mechanical damage, presented the best quality standards. The fruits suffered only one kind of damage, when packaged, presented satisfactory pattern compared to the fruits without package and not exposed to any mechanical damages. Applying the two kinds of damages, the LDPE packaging was not adequate to decrease the metabolic rate of these fruits, making them unfit for marketing.

Produção de amilases de Rhizopus microsporus var. oligosporus e hidrólise enzimática do bagaço de mandioca visando a produção de etanol por Saccharomyces cerevisiae

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Produção de amilases de Rhizopus microsporus var. oligosporus e hidrólise enzimática do bagaço de mandioca visando a produção de etanol por Saccharomyces cerevisiae

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Biotic versus abiotic pollination in Oeceoclades maculata (Lindl.) Lindl. (Orchidaceae)

Geographical variation in the reproductive biology of widespread species often occurs at their distributional boundaries. We sought to determine whether such variation has occurred in an invasive orchid, Oeceoclades maculata, across its naturalized range. We compared its reproductive biology in a Brazilian population with that published for a population on the Caribbean island of Puerto Rico. In the state of Sao Paulo, O. maculata flowers between December and February, at the height of the rainy season. Similar fruit sets were observed in manual self (76%) and cross (70.4%) pollination treatments. The fruit set of plants protected from both pollinators and rainfall was 6.1%, whereas plants exposed only to rainfall had a fruit set of 41.4%, slightly less than the controls (48.3%). Like the Puerto Rico population, reproduction is primarily through rain-assisted autogamy, but unlike observations made on the island, outcrossing can eventually occur. We observed two butterfly species (Heliconius ethilla narcaea and Heliconius erato phyllis) pollinating O. maculata. Secretory epidermal cells and trichomes of the spur lumen produced 0.7 mu L of 25% (sucrose equivalents) nectar per flower each morning, which was stored in a dilated basal portion of the spur and reabsorbed by the afternoon. Thus, geographical variation in reproductive biology exists across the broad invasive range of O. maculata.

Production of a xylose-stimulated beta-glucosidase and a cellulase-free thermostable xylanase by the thermophilic fungus Humicola brevis var. thermoidea under solid state fermentation

Humicola brevis var. thermoidea cultivated under solid state fermentation in wheat bran and water (1:2 w/v) was a good producer of beta-glucosidase and xylanase. After optimization using response surface methodology the level of xylanase reached 5,791.2 +/- A 411.2 U g(-1), while beta-glucosidase production was increased about 2.6-fold, reaching 20.7 +/- A 1.5 U g(-1). Cellulase levels were negligible. Biochemical characterization of H. brevis beta-glucosidase and xylanase activities showed that they were stable in a wide pH range. Optimum pH for beta-glucosidase and xylanase activities were 5.0 and 5.5, respectively, but the xylanase showed 80 % of maximal activity when assayed at pH 8.0. Both enzymes presented high thermal stability. The beta-glucosidase maintained about 95 % of its activity after 26 h in water at 55 A degrees C, with half-lives of 15.7 h at 60 A degrees C and 5.1 h at 65 A degrees C. The presence of xylose during heat treatment at 65 A degrees C protected beta-glucosidase against thermal inactivation. Xylanase maintained about 80 % of its activity after 200 h in water at 60 A degrees C. Xylose stimulated beta-glucosidase activity up to 1.7-fold, at 200 mmol L-1. The notable features of both xylanase and beta-glucosidase suggest that H. brevis crude culture extract may be useful to compose efficient enzymatic cocktails for lignocellulosic materials treatment or paper pulp biobleaching.

Effects of Natural Radiation, Photosynthetically Active Radiation and Artificial Ultraviolet Radiation-B on the Chloroplast Organization and Metabolism of Porphyra acanthophora var. brasiliensis (Rhodophyta, Bangiales)

We undertook a study of Porphyra acanthophora var. brasiliensis to determine its responses under ambient conditions, photosynthetically active radiation (PAR), and PAR+UVBR (ultraviolet radiation-B) treatment, focusing on changes in ultrastructure, and cytochemistry. Accordingly, control ambient samples were collected in the field, and two different treatments were performed in the laboratory. Plants were exposed to PAR at 60 mu mol photons m(-2) s(-1) and PAR+UVBR at 0.35 W m(-2) for 3 h per day during 21 days of in vitro cultivation. Confocal laser scanning microscopy analysis of the vegetative cells showed single stellate chloroplast in ambient and PAR samples, but in PAR+UVBR-exposed plants, the chloroplast showed alterations in the number and form of arms. Under PAR+UVBR treatment, the thylakoids of the chloroplasts were disrupted, and an increase in the number of plastoglobuli was observed, in addition to mitochondria, which appeared with irregular, disrupted morphology compared to ambient and PAR samples. After UVBR exposure, the formation of carpospores was also observed. Plants under ambient conditions, as well as those treated with PAR and PAR+UVBR, all showed different concentrations of enzymatic response, including glutathione peroxidase and reductase activity. In summary, the present study demonstrates that P. acanthophora var. brasiliensis shows the activation of distinct mechanisms against natural radiation, PAR and PAR+UVBR.

Evaluation of the genetic diversity of Histoplasma capsulatum var. capsulatum isolates from north-eastern Brazil

Since the beginning of the HIV epidemic, there has been a significant increase in the number of histoplasmosis cases in Ceara, a state in north-east Brazil. The lack of epidemiological data on the genotypes circulating in the north-east region shows the importance of more detailed studies on the molecular epidemiology of Histoplasma capsulatum var. capsulatum in this region. Different molecular techniques have been used to better characterize the genetic profile of H. capsulatum var. capsulatum strains. The aim of this study was to analyse the genetic diversity of H. capsulatum var. capsulatum isolates in Fortaleza, the capital of Ceara, through the sequencing of the internal transcribed spacer (ITS)1-5.8S-ITS2 region, and establish the molecular profile of these isolates, along with strains from south-east Brazil, by RAPD analysis, featuring the different clusters in those regions. The isolates were grouped into two clusters. Cluster 1 included strains from the south-east and north-east regions with separation of isolates into three distinct subgroups (subgroups 1a, 1 b and 1 c). Cluster 2 included only samples from north-east Brazil. Sequencing of the ITS1 -5.8S-ITS2 region allowed the detection of two major clades, which showed geographical correlation between them and their subgroups. Therefore, it can be concluded that the H. capsulatum var. capsulatum isolates from Ceara have a high degree of genetic polymorphism. The molecular data also confirm that populations of this fungus are composed of different genotypes in Brazil and worldwide.

Megabenthic communities in the waters around Svalbard during POLARSTERN cruise ARK-VIII/2 (EPOS II)

Composition and distribution of megabenthic communities around Svalbard were investigated in June/July 1991 with 20 Agassiz trawl and 5 bottom trawl hauls in depths between 100 and 2100 m. About 370 species, ranging from sponges to fish, were identified in the catches. Species numbers per station ranged from 21 to 86. Brittle stars, such as Ophiacantha bidentata, Ophiura sarsi and Ophiocten sericeum, were most important in terms of constancy and relative abundance in the catches. Other prominent faunal elements were eunephthyid alcyonarians, bivalves, shrimps, sea stars and fish (Gadidae, Zoarcidae, Cottidae). Multivariate analyses of the species and environmental data sets showed that the spatial distribution of the megabenthos was characterized by a pronounced depth zonation: abyssal, bathyal, off-shore shelf and fjordic communities were discriminated. However, a gradient in sediment properties, especially the organic carbon content, seemed to superimpose on the bathymetric pattern. Both main factors are interpreted as proxies of the average food availability, which is, hence, suggested to have the strongest influence in structuring megabenthic communities off Svalbard.

Nitrógeno prontamente asimilable : efecto sobre la velocidad de fermentación del jugo de uva (Vitis vinifera L.)

El propósito de este trabajo fue evaluar la incidencia de la concentración de nitrógeno prontamente asimilable (NPA) sobre la velocidad y duración de la fermentación alcohólica de los mostos de uva. El experimento se diseñó con tres tratamientos (A=testigo; B=agregado de PO4H(NH4)2 50 mg/L; C=agregado de PO4H(NH4)2 100 mg/L) y cuatro repeticiones. Se realizaron microvinificaciones con jugo de uva pasteurizado var. Chardonnay, inoculado con Saccharomyces cerevisiae cepa FCA 32. La fermentación se condujo a 25 °C. La concentración de NPA fue medida por titulación en medio formol. La velocidad de fermentación fue determinada por pérdida de peso. La velocidad máxima de fermentación se alcanzó al tercer día. Existen diferencias significativas entre la velocidad máxima alcanzada por el testigo y por los tratamientos B y C pero no hay diferencias significativas entre las velocidades máximas alcanzadas por los tratamientos B y C. La velocidad máxima de fermentación alcanzada por el tratamiento B (agregado de 50 mg/L de PO4H(NH4)2) fue 57 % superior respecto del testigo, mientras que el tratamiento C (agregado de 100 mg/L de PO4H(NH4)2) fue 53 % superior respecto del mismo testigo La velocidad máxima de fermentación aumentó con la adición de nitrógeno, pero no se observan diferencias entre las distintas dosis empleadas. La duración media de la fermentación resulta significativamente diferente para los tres tratamientos: 9.25 días para el testigo, 7.5 días para el tratamiento B y 6.25 días para el tratamiento C. El agregado de PO4H(NH4)2 disminuye la duración de la fermentación en las condiciones de trabajo. La duración de la fermentación del tratamiento B (agregado de 50 mg/L de PO4H(NH4)2) fue del 81 % respecto del testigo 100 %, mientras que el tratamiento C (agregado de 100 mg/L de PO4H(NH4)2) fue del 67 %, respecto del mismo testigo.

Bioensayo de germinación de Lactuca sativa (L.) : determinación de calidad de agua en represas para riego

En el norte de la provincia de Entre Ríos se ha desarrollado un modelo de producción de arroz basado en el uso de agua superficial mediante la construcción de presas de tierra: existen más de 60 con superficies variables. El uso de agroquímicos en esa zona podría introducir un factor de contaminación en el suelo y en el agua de las represas. El objetivo de este trabajo fue determinar la calidad del agua de 19 represas para riego del centro norte de Entre Ríos, mediante bioensayos de germinación de Lactuca sativa var. mantecosa. Los bioensayos se realizaron en cajas de Petri, con papel de filtro en la base humedecido con 3 ml de agua de la muestra correspondiente. Se sembraron 20 semillas por caja, distribuyéndose los tratamientos en bloques al azar con 4 repeticiones, en cámara de germinación con alternancia de luz y oscuridad. Se registró el porcentaje de germinación y la longitud promedio de la raíz; se calculó un índice de germinación. El porcentaje de germinación promedio de los tratamientos fue de 96,07% y de 97,9% en el testigo. El índice de germinación en todos los casos fue superior al 60% y no se detectó toxicidad en el agua proveniente de las distintas represas.

Actividad tóxica del aceite esencial de laurel y del cineol sobre Brevicoryne brassicae L. en repollo

El objetivo del presente trabajo fue estudiar la actividad tóxica del aceite esencial de laurel (Laurus nobilis L.) y del cineol, monoterpeno cíclico considerado un aleloquímico puro, sobre Brevicoryne brassicae L. en repollo (Brassica oleracea var. capitata L.). Las concentraciones de aceite esencial utilizadas fueron: 1; 1,5; 2 y 3%, y para cineol: 0,5; 1,5 y 2,5%, ambas formuladas en solución acuosa con 2% de oleato de polietilenglicol como emulsionante. Se utilizaron dos técnicas de aplicación: papeles impregnados y pulverización directa. A las 24 horas del tratamiento se evaluó el porcentaje de mortalidad. Los resultados se analizaron por ANOVA de dos vías y Test de Tukey. Se encontraron diferencias significativas para los productos evaluados en todas las concentraciones ensayadas. Las técnicas de aplicación no dieron diferencias significativas. La mayor mortalidad correspondió a 52% para el aceite esencial de laurel y 27,5% para cineol a las mayores concentraciones en ambos casos. Se concluye que los productos ensayados podrían ser una herramienta para el Manejo Integrado de Plagas para control de áfidos en cultivos hortícolas.

Uso de marcadores morfológicos, bioquímicos y moleculares SRAP para diferenciar variedades de Cynara cardunculus L. (Asteraceae)

Dentro del complejo primario de Cynara cardunculus L. se encuentran diversas variedades botánicas: var. scolymus (alcaucil), var. altilis (cardo cultivado) y var. sylvestris (cardo silvestre). A lo largo de la historia del mejoramiento, la caracterización de los materiales ha evolucionado desde el uso de caracteres morfológicos hasta los modernos análisis moleculares, pasando por los marcadores bioquímicos. El objetivo del presente trabajo fue comparar la utilidad de los marcadores morfológicos, bioquímicos y moleculares para la caracterización de materiales pertenecientes a tres variedades botánicas de Cynara cardunculus. Tres cultivares de la var. scolymus, dos de la var. altilis y dos de la var. sylvestris fueron caracterizadas por variables morfovegetativas, proteínas de reserva y por marcadores moleculares a través de la técnica SRAP. Estas metodologías permitieron discriminar dos grupos: uno incluyendo las variedades de cardo cultivado y silvestre, y el otro, las variedades del alcaucil. Los datos moleculares y morfológicos permitieron además diferenciar los cultivares evaluados de la var. scolymus. Se concluye que los marcadores analizados son útiles para la caracterización intravarietal e intervarietal en programas de mejoramiento.

Variability in the indument of Phaseolus vulgaris var. aborigineus (Fabaceae)

The indument of Phaseolus vulgaris L. var. aborigineus (Burkart) Baudet, a native bean from Argentina, potential source for improvement of cultivated varieties, was compared between different genetic lines. Seeds collected from wild, weedy forms and presumptive hybrid individuals in two distant latitudes were sown in two sites of Buenos Aires University Campus (field and greenhouse). Two type of trichomes were found, which differenciate in length and shape. The pubescence density was significantly different between wild specimens from different latitudes and between culture sites for plants of a same procedence. This character could be involved in physical defences of these plants.

Diatom biostratigraphy of sediments from the Kerguelen Plateau, Southern Ocean

The biostratigraphic distribution and abundance of lower Oligocene to Pleistocene diatoms is documented from Holes 747A, 747B, 748B, 749B, and 751A drilled during Ocean Drilling Program Leg 120 on the Kerguelen Plateau in the southeast Indian Ocean. The occurrence of middle and upper Eocene diatoms is also documented, but these are rare and occur in discrete intervals. The recovery of several Oligocene to Pleistocene sections with minimal coring gaps, relatively good magnetostratigraphic signatures, and mixed assemblages of both calcareous and siliceous microfossils makes the above four Leg 120 sites important biostratigraphic reference sections for the Southern Ocean and Antarctic continent. A high-resolution diatom zonation divides the last 36 m.y. into 45 zones and subzones. This zonation is built upon an existing biostratigraphic framework developed over the past 20 yr of Southern Ocean/Antarctic deep-sea coring and drilling. After the recent advances from diatom biostratigraphic studies on sediments from Legs 113, 114, 119, and 120, a zonal framework for the Southern Ocean is beginning to stabilize. The potential age resolution afforded by the high-diversity diatom assemblages in this region ranks among the highest of all fossil groups. In addition to the 46 datum levels that define the diatom zones and subzones, the approximate stratigraphic level, age, and magnetic anomaly correlative of more than 150 other diatom datums are determined or estimated. These total 73 datum levels for the Pliocene-Pleistocene, 67 for the Miocene, and 45 for the Oligocene. Greater stratigraphic resolution is possible as the less common and poorly documented species become better known. This high-resolution diatom stratigraphy, combined with good to moderately good magnetostratigraphic control, led to the recognition of more than 10 intervals where hiatuses dissect the Oligocene-Pleistocene section on the Kerguelen Plateau. We propose 12 new diatom taxa and 6 new combination