11-Oxoaerothionin isolated from the marine sponge Aplysina fistularis shows anti-inflammatory activity in LPS-stimulated macrophages

Marine sponges of the order Verongida are a rich source of biologically active bromotyrosine-derived secondary metabolites. However, none of these compounds are known to display anti-inflammatory activity. In the present investigation, we report the anti-inflammatory effects of 11-oxoaerothionin isolated from the Verongida sponge Aplysina fistularis. When RAW264.7 cells and primary macrophages were preincubated with 11-oxoaerothionin and stimulated with LPS (lipopolysaccharide), a concentration-dependent inhibition of iNOS (inducible nitric oxide synthase) protein and NO2- (Nitrite) production were observed. The same effect was observed when proinflammatory cytokines and PGE(2) (Prostaglandin E2) production was evaluated. In summary, we demonstrated that in the presence of LPS, 11-oxoaerothionin suppresses NO2 and iNOS expression as well as inflammatory cytokines and PGE(2).

HEPATOTOXICITY ASSESSMENT OF THE AZO DYES DISPERSE ORANGE 1 (DO1), DISPERSE RED 1 (DR1,) AND DISPERSE RED 13 (DR13) IN HEPG2 CELLS

During the dyeing process in baths approximately 10 to 15% of the dyes used are lost and reach industrial effluents, thus polluting the environment. Studies showed that some classes of dyes, mainly azo dyes and their by-products, exert adverse effects on humans and local biota, since the wastewater treatment systems and water treatment plants were found to be ineffective in removing the color and reducing toxicity of some dyes. In the present study, the toxicity of the azo dyes disperse orange 1 (DO1), disperse red 1 (DR1), and disperse red 13 (DR13) was evaluated in HepG2 cells grown in monolayers or in three dimensional (3D) culture. Hepatotoxicity of the dyes was measured using 3-(4,5-dimethylthiazol-2yl)2,5-diphenyltetrazolium (MTT) and cell counting kit 8 (CCK-8) assays after 24, 48, and 72 h of incubation of cells with 3 different concentrations of the azo dyes. The dye DO1 only reduced the mitochondrial activity in HepG2 cells grown in a monolayer after 72 h incubation, while the dye DR1 showed this deleterious effect in both monolayer and 3D culture. In contrast, dye DR13 decreased the mitochondrial activity after 24, 48, and 72 h of exposure in both monolayer and 3D culture. With respect to dehydrogenase activity, only the dye DR13 diminished the activity of this enzyme after 72 h of exposure in both monolayer and 3D culture. Our results clearly demonstrated that exposure to the studied dyes induced cytotoxicity in HepG2 cells.

Soil-mediated effects on potential Euterpe edulis (Arecaceae) fruit and palm heart sustainable management in the Brazilian Atlantic Forest

Euterpe edulis is an endangered species due to palm heart overharvesting, the most important non-timber forest product of the Brazilian Atlantic Forest, and fruit exploitation has been introduced as a low impacting alternative. However, E. edulis is a keystone species for frugivores birds, and even the impact of fruit exploitation needs to be better investigated. Since this species occurs over contrasting habitats, the establishment of site-specific standards and limits for exploitation may also be essential to achieve truly sustainable management. In this context, we sought to investigate how soil chemical composition would potentially affect E. edulis (Arecaceae) palm heart and fruit exploitation considering current standards of management. We studied natural populations found in Restinga Forest and Atlantic Rainforest remnants established within Natural Reserves of Sao Paulo State, SE Brazil, where 10.24 ha permanent plots, composed of a grid of 256 subplots (20 m x 20 m), were located. In each of these subplots, we evaluated soil chemical composition and diameter at breast height of E. edulis individuals. Additionally, we evaluated fruit yield in 2008 and 2009 in 20 individuals per year. The Atlantic Rainforest population had a much higher proportion of larger diameter individuals than the population from the Restinga Forest, as a result of habitat-mediated effects, especially those related to soil. Sodium and potassium concentration in Restinga Forest soils, which have strong negative and positive effect on palm growth, respectively, played a key role in determining those differences. Overall, the number of fruits that could be exploited in the Atlantic Rainforest was four times higher than in Restinga Forest. If current rules for palm heart and fruit harvesting were followed without any restriction to different habitats, Restinga Forest populations are under severe threat, as this study shows that they are not suitable for sustainable management of both fruits and palm heart. Hence, a habitat-specific approach of sustainable management is needed for this species in order to respect the demographic and ecological dynamics of each population to be managed. These findings suggest that any effort to create general management standards of low impacting harvesting may be unsuccessful if the species of interest occur over a wide range of ecosystems. (C) 2012 Elsevier B.V. All rights reserved.

Microbial transformation of the sesquiterpene lactone tagitinin C by the fungus Aspergillus terreus

The biotransformation of the sesquiterpene lactone tagitinin C by the fungus Aspergillus terreus MT 5.3 yielded a rare derivative that was elucidated by spectrometric methods. The fungus led to the formation of a different product through an unusual epoxidation reaction between C4 and C5, formation of a C3,C10 ether bridge, and a methoxylation of the C1 of tagitinin C. The chemical structure of the product, namely 1 beta-methoxy-3 alpha-hydroxy-3,10 beta-4,5 alpha-diepoxy-8 beta-isobutyroyloxygermacr-11(13)-en-6 alpha,12-olide, is the same as that of a derivative that was recently isolated from the flowers of a Brazilian population of Mexican sunflower (Tithonia diversifolia), which is the source of the substrate tagitinin C. The in vitro cytotoxic activity of the substrate and the biotransformed product were evaluated in HL-60 cells using an MTT assay, and both compounds were found to be cytotoxic. We show that soil fungi may be useful in the biotransformation of sesquiterpene lactones, thereby leading to unusual changes in their chemical structures that may preserve or alter their biological activities, and may also mimic plant biosynthetic pathways for production of secondary metabolites.

Fishes of Paranagua Estuarine Complex, South West Atlantic

The objective of this work was to present an updated checklist of the currently known fishes in the Paranagua Estuarine Complex (PEC) and provides comments on conservation status for the treated species. We used a large dataset derived from a pool of studies which have been conducted within there along the last 30 years. Each study was based on monthly samplings and conducted in several estuarine habitat; thus, the pool covers practically all estuarine habitats and takes into account the seasonal cycle in the system. The PEC ichthyofauna represents a mixture between that fauna typical from the tropical Brazilian coast and that with affinities of temperate Argentinean and Uruguayan zones. The PEC harbors a rich fish fauna of 213 species, inserted in the families that are common along the Brazilian coast. Only a minor part (8%) of the PEC fish fauna was evaluated as regards the conservation status, mostly because of the lack of basic biological and ecological information for most species. Despite part of the among-estuaries differences are due to different and incomplete sampling efforts, the richness in the PEC is surprisingly higher than other systems in Brazil and around world, which emphasize the importance of the region for global biodiversity conservation.

Differential seed germination of a keystone palm (Euterpe edulis) dispersed by avian frugivores

FAPESP (BIOTA Program) [2007/03392-6, 2010/04927-3]

Systematic UPLC-ESI-MS/MS Study on the Occurrence of Staurosporine and Derivatives in Associated Marine Microorganisms from Eudistoma vannamei

Former bioactivity-guided analysis of the marine invertebrate Eudistoma vannamei led to the isolation of staurosporine derivatives, which revealed strong cytotoxic activity against several human cancer cell lines. The occurrence of such alkaloids in E. vannamei may be correlated to the presence of associated biota, such as Streptomyces bacteria. In agreement to this hypothesis, marine microorganisms associated with E. vannamei were recovered and cultured, leading to a total of 84 isolated bacterial strains. Gas phase fragmentation reactions of staurosporine and derivatives were systematically studied and the analyzed results further supported by computational chemistry studies. The resulting fragment patterns were used to search for the presence of different derivatives in extracts of isolated microorganisms, thereby using LC-MS/MS analysis in MRM mode. These results evidenced that one isolated Streptomyces sp. was able to generate staurosporine, while none of the hydroxy-7-oxo derivatives were detected. Finally, significant cytotoxic activity against human cancer lines was observed for one of the extracts.

Composição, diversidade e distribuição geográfica de plantas vasculares de uma Floresta Ombrófila Densa Atlântica do Sudeste do Brasil

In the Montane and Submontane Rain Forest of the Carlos Botelho State Park - PECB (ca. 37,000 ha) the composition, richness and geographical distribution of native, vascular forest species was evaluated. The analysis of 1143 species of 140 families supported the pattern found for other forests of Eastern Brazil, showing high species richness of Myrtaceae (85 species), Orchidaceae (81), Fabaceae (57), Asteraceae, Melastomataceae (54), Lauraceae (53), Rubiaceae (51), Bromeliaceae (43), Piperaceae (30) and Solanaceae (25), besides ferns (123). The most species-rich genera were Eugenia (34), Ocotea (26), Leandra, Myrcia, Vriesea (18), Piper, Solanum (16), Miconia (14), Mollinedia (13), and Peperomia (12). The richness and composition varied greatly among life forms, as well as the number of families represented in each one of them (only Rubiaceae had species in all life forms, except parasites). Trees had the largest contribution of total richness (39.1%), a value that represented more than 20% of the species listed for the whole Atlantic Forest of Southeastern Brazil. Trees were followed by epiphytes (22.4%), herbs (18.4%), shrubs (10.1%), lianas (9.1%), and parasites (0.9%). The overall richness and composition of life forms was quite close to other neotropical forests (e.g. high contribution of ferns among epiphytes), although some life forms remain undersampled in the PECB (mainly herbs, lianas and epiphytes). The occurrence of species endemic to the Atlantic Forest was pronounced (65%), with a predominance of species restricted to the Southern Atlantic Forest (43%). Pantropical species were rare (2%), being more common among ferns. Myrtaceae and Melastomataceae were the families with greater number and proportion of endemic species.

Shifts in phylogenetic diversity of archaeal communities in mangrove sediments at different sites and depths in southeastern Brazil

This study focused on the structure and composition of archaeal communities in sediments of tropical mangroves in order to obtain sufficient insight into two Brazilian sites from different locations (one pristine and another located in an urban area) and at different depth levels from the surface. Terminal restriction fragment length polymorphism (T-RFLP) of PCR-amplified 16S rRNA gene fragments was used to scan the archaeal community structure, and 16S rRNA gene clone libraries were used to determine the community composition. Redundancy analysis of T-RFLP patterns revealed differences in archaeal community structure according to location, depth and soil attributes. Parameters such as pH, organic matter, potassium and magnesium presented significant correlation with general community structure. Furthermore, phylogenetic analysis revealed a community composition distributed differently according to depth where, in shallow samples, 74.3% of sequences were affiliated with Euryarchaeota and 25.7% were shared between Crenarchaeota and Thaumarchaeota, while for the deeper samples, 24.3% of the sequences were affiliated with Euryarchaeota and 75.7% with Crenarchaeota and Thaumarchaeota. Archaeal diversity measurements based on 16S rRNA gene clone libraries decreased with increasing depth and there was a greater difference between depths (<18% of sequences shared) than sites (>25% of sequences shared). Taken together, our findings indicate that mangrove ecosystems support a diverse archaeal community; it might possibly be involved in nutrient cycles and are affected by sediment properties, depth and distinct locations. (C) 2012 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Endophytic fungi from Vitis labrusca L. ('Niagara Rosada') and its potential for the biological control of Fusarium oxysporum

We investigated the diversity of endophytic fungi found on grape (Vitis labrusca cv. Niagara Rosada) leaves collected from Salesopolis, SP, Brazil. The fungi were isolated and characterized by amplified ribosomal DNA restriction analysis, followed by sequencing of the ITS1-5.8S-ITS2 rDNA. In addition, the ability of these endophytic fungi to inhibit the grapevine pathogen Fusarium oxysporum f. sp herbemontis was determined in vitro. We also observed that the climatic factors, such as temperature and rainfall, have no effect on the frequency of infection by endophytic fungi. The endophytic fungal community that was identified included Aporospora terricola, Aureobasidium pullulans, Bjerkandera adusta, Colletotrichum boninense, C. gloeosporioides, Diaporthe helianthi, D. phaseolorum, Epicoccum nigrum, Flavodon flavus, Fusarium subglutinans, F. sacchari, Guignardia mangiferae, Lenzites elegans, Paraphaeosphaeria pilleata, Phanerochaete sordida, Phyllosticta sp, Pleurotus nebrodensis, Preussia africana, Tinctoporellus epiniltinus, and Xylaria berteri. Among these isolates, two, C. gloeosporioides and F. flavus, showed potential antagonistic activity against F. oxysporum f. sp herbemontis. We suggest the involvement of the fungal endophyte community of V. labrusca in protecting the host plant against pathogenic Fusarium species. Possibly, some endophytic isolates could be selected for the development of biological control agents for grape fungal disease; alternatively, management strategies could be tailored to increase these beneficial fungi.

Application of MALDI-MS analysis of Rainforest chemodiversity: a keystone for biodiversity conservation and sustainable use

FAPESP/BIOTA

Systematic UPLC-ESI-MS/MS study on the occurrence of staurosporine and derivatives in associated marine microorganisms from Eudistoma vannamei

Former bioactivity-guided analysis of the marine invertebrate Eudistoma vannamei led to the isolation of staurosporine derivatives, which revealed strong cytotoxic activity against several human cancer cell lines. The occurrence of such alkaloids in E. vannamei may be correlated to the presence of associated biota, such as Streptomyces bacteria. In agreement to this hypothesis, marine microorganisms associated with E. vannamei were recovered and cultured, leading to a total of 84 isolated bacterial strains. Gas phase fragmentation reactions of staurosporine and derivatives were systematically studied and the analyzed results further supported by computational chemistry studies. The resulting fragment patterns were used to search for the presence of different derivatives in extracts of isolated microorganisms, thereby using LC-MS/MS analysis in MRM mode. These results evidenced that one isolated Streptomyces sp. was able to generate staurosporine, while none of the hydroxy-7-oxo derivatives were detected. Finally, significant cytotoxic activity against human cancer lines was observed for one of the extracts.

Invasion of the dinoflagellate Ceratium furcoides (Levander) Langhans 1925 at tropical reservoir and its relation to environmental variables

Dinoflagellates of the genus Ceratium are chiefly marine but there are rare occurrences in freshwater. In this study we analyze the invasion and progressive establishment of Ceratium furcoides, an exotic species, in the Furnas Reservoir. Samples were taken at 36 points in the reservoir, during the months of March, June, September and December, 2007. Measurements of some physical and chemical variables were simultaneously performed at each site. The occurrence of C. furcoides was registered at 20 sites, with densities varying between 0.57 and 28,564,913.0 ind.m-3. Blooms of this species were recorded in points which were classified as mesotrophic, coinciding with the places receiving high amounts of untreated domestic sewage. C. furcoides density was correlated with temperature, nutrients (nitrate and nitrite) and water electric conductivity. The highest density was recorded in June when temperature was low. The presence of Ceratium furcoides in the reservoir apparently has not yet affected the reservoir water quality or other plankton communities. However, if it becomes fully established it could perhaps become a problem in the reservoir or even to spread out to other reservoirs in Rio Grande basin.

Fishes of Paranaguá Estuarine Complex, South West Atlantic

The objective of this work was to present an updated checklist of the currently known fishes in the Paranaguá Estuarine Complex (PEC) and provides comments on conservation status for the treated species. We used a large dataset derived from a pool of studies which have been conducted within there along the last 30 years. Each study was based on monthly samplings and conducted in several estuarine habitat; thus, the pool covers practically all estuarine habitats and takes into account the seasonal cycle in the system. The PEC ichthyofauna represents a mixture between that fauna typical from the tropical Brazilian coast and that with affinities of temperate Argentinean and Uruguayan zones. The PEC harbors a rich fish fauna of 213 species, inserted in the families that are common along the Brazilian coast. Only a minor part (8%) of the PEC fish fauna was evaluated as regards the conservation status, mostly because of the lack of basic biological and ecological information for most species. Despite part of the among-estuaries differences are due to different and incomplete sampling efforts, the richness in the PEC is surprisingly higher than other systems in Brazil and around world, which emphasize the importance of the region for global biodiversity conservation.

Life history of three catfish species (Siluriformes: Ariidae) from southeastern Brazil

This study evaluated the spatio-temporal distribution, reproduction and diet of the catfishes Genidens genidens, G. barbus and Aspistor luniscutis in Caraguatatuba Bay. Their sizes were recorded and the sex and reproductive stage identified. The abundance was compared between areas (South and North) and among months (August 2003 - October 2004). The species had different spatial distributions, allowing them to coexist. The temporal distribution reflected their tendencies to migrate in the reproductive period, as evidenced by the dominance of small immature individuals in the bay. G. genidens tended to reproduce in winter, and A. luniscutis in spring. The diet of G. genidens consisted of crustaceans, mollusks (shells), fish scales, ostracods, and bivalve siphons. G. barbus consumed a high proportion of mysids, followed by fish (bones and scales). For A. luniscutis, the diet was based on fish scales and crustaceans. High quantities of particulate organic matter were observed in the diet of all three species, as previously known for estuarine catfishes. The consumption of fish scales may reflect a lepidophagic habit. A small overlap was observed among the diets, reflecting differences in their environments as well as in the proportions of each item ingested.