Brown algal phlorotannins: Improving and applying chemical methods

Phlorotannins are the least studied group of tannins and are found only in brown algae. Hitherto the roles of phlorotannins, e.g. in plant-herbivore interactions, have been studied by quantifying the total contents of the soluble phlorotannins with a variety of methods. Little attention has been given to either quantitative variation in cell-wall-bound and exuded phlorotannins or to qualitative variation in individual compounds. A quantification procedure was developed to measure the amount of cell-wall-bound phlorotannins. The quantification of soluble phlorotannins was adjusted for both large- and small-scale samples and used to estimate the amounts of exuded phlorotannins using bladder wrack (Fucus vesiculosus) as a model species. In addition, separation of individual soluble phlorotannins to produce a phlorotannin profile from the phenolic crude extract was achieved by high-performance liquid chromatography (HPLC). Along with these methodological studies, attention was focused on the factors in the procedure which generated variation in the yield of phlorotannins. The objective was to enhance the efficiency of the sample preparation procedure. To resolve the problem of rapid oxidation of phlorotannins in HPLC analyses, ascorbic acid was added to the extractant. The widely used colourimetric method was found to produce a variation in the yield that was dependent upon the pH and concentration of the sample. Using these developed, adjusted and modified methods, the phenotypic plasticity of phlorotannins was studied with respect to nutrient availability and herbivory. An increase in nutrients decreased the total amount of soluble phlorotannins but did not affect the cell-wall-bound phlorotannins, the exudation of phlorotannins or the phlorotannin profile achieved with HPLC. The presence of the snail Thedoxus fluviatilis on the thallus induced production of soluble phlorotannins, and grazing by the herbivorous isopod Idotea baltica increased the exudation of phlorotannins. To study whether the among-population variations in phlorotannin contents arise from the genetic divergence or from the plastic response of algae, or both, algae from separate populations were reared in a common garden. Genetic variation among local populations was found in both the phlorotannin profile and the content of total phlorotannins. Phlorotannins were also genetically variable within populations. This suggests that local algal populations have diverged in their contents of phlorotannins, and that they may respond to natural selection and evolve both quantitatively and qualitatively.

A spatial modelling framework for assessing climate change impacts on freshwater ecosystems: Response of brown trout (Salmo trutta L.) biomass to warming water temperature

Mountain regions worldwide are particularly sensitive to on-going climate change. Specifically in the Alps in Switzerland, the temperature has increased twice as fast than in the rest of the Northern hemisphere. Water temperature closely follows the annual air temperature cycle, severely impacting streams and freshwater ecosystems. In the last 20 years, brown trout (Salmo trutta L) catch has declined by approximately 40-50% in many rivers in Switzerland. Increasing water temperature has been suggested as one of the most likely cause of this decline. Temperature has a direct effect on trout population dynamics through developmental and disease control but can also indirectly impact dynamics via food-web interactions such as resource availability. We developed a spatially explicit modelling framework that allows spatial and temporal projections of trout biomass using the Aare river catchment as a model system, in order to assess the spatial and seasonal patterns of trout biomass variation. Given that biomass has a seasonal variation depending on trout life history stage, we developed seasonal biomass variation models for three periods of the year (Autumn-Winter, Spring and Summer). Because stream water temperature is a critical parameter for brown trout development, we first calibrated a model to predict water temperature as a function of air temperature to be able to further apply climate change scenarios. We then built a model of trout biomass variation by linking water temperature to trout biomass measurements collected by electro-fishing in 21 stations from 2009 to 2011. The different modelling components of our framework had overall a good predictive ability and we could show a seasonal effect of water temperature affecting trout biomass variation. Our statistical framework uses a minimum set of input variables that make it easily transferable to other study areas or fish species but could be improved by including effects of the biotic environment and the evolution of demographical parameters over time. However, our framework still remains informative to spatially highlight where potential changes of water temperature could affect trout biomass. (C) 2015 Elsevier B.V. All rights reserved.-

Evaluation of a revision of the BSPcast decision support system for control of brown spot of pear

Control of brown spot of pear requires fungicide treatments of pear trees during the growing season. Scheduling fungicide sprays with the Brown spot of pear forecasting system (BSPcast) provides significantfungicide savings but does not increase the efficacy of disease control. Modifications in BSPcast wereintroduced in order to increase system performance. The changes consisted of: (1) the use of a daily infectionrisk (Rm≥0.2) instead of the 3-day cumulative risk (CR≥0.4) to guide the fungicide scheduling, and (2) theinclusion of the effect of relative humidity during interrupted wetness periods. Trials were performed during2 years in an experimental pear orchard in Spain. The modifications introduced did not result in increaseddisease control efficacy, compared with the original BSPcast system. In one year, no reduction in the numberof fungicide applications was obtained using the modified BSPcast system in comparison to the original system, but in the second year the number of treatments was reduced from 15 to 13. The original BSPcast model overestimated the daily infection risk in 6.5% of days with wetness periods with low relative humidity during the wetness interruption, and in these cases the modified version was more adequate

Increased diversity of egg-associated bacteria on brown trout (Salmo trutta) at elevated temperatures.

The taxonomic composition of egg-associated microbial communities can play a crucial role in the development of fish embryos. In response, hosts increasingly influence the composition of their associated microbial communities during embryogenesis, as concluded from recent field studies and laboratory experiments. However, little is known about the taxonomic composition and the diversity of egg-associated microbial communities within ecosystems; e.g., river networks. We sampled late embryonic stages of naturally spawned brown trout at nine locations within two different river networks and applied 16S rRNA pyrosequencing to describe their bacterial communities. We found no evidence for a significant isolation-by-distance effect on the composition of bacterial communities, and no association between neutral genetic divergence of fish host (based on 11 microsatellites) and phylogenetic distances of the composition of their associated bacterial communities. We characterized core bacterial communities on brown trout eggs and compared them to corresponding water samples with regard to bacterial composition and its presumptive function. Bacterial diversity was positively correlated with water temperature at the spawning locations. We discuss this finding in the context of the increased water temperatures that have been recorded during the last 25 years in the study area.

Long-term treatment with insulin induces apoptosis in brown adipocytes: role of oxidative stress

Trying to define the precise role played by insulin regulating the survival of brown adipocytes, we have used rat fetal brown adipocytes maintained in primary culture. The effect of insulin on apoptosis and the mechanisms involved were assessed. Different from the known effects of insulin as a survival factor, we have found that long-term treatment (72 h) with insulin induces apoptosis in rat fetal brown adipocytes. This process is dependent on the phosphatidylinositol 3-kinase/mammalian target of rapamycin/p70 S6 kinase pathway. Short-term treatment with the conditioned medium from brown adipocytes treated with insulin for 72 h mimicked the apoptotic effect of insulin. During the process, caspase 8 activation, Bid cleavage, cytochrome c release, and activation of caspases 9 and 3 are sequentially produced. Treatment with the caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp (Z-VAD), prevents activation of this apoptotic cascade. The antioxidants, ascorbic acid and superoxide dismutase, also impair this process of apoptosis. Moreover, generation of reactive oxygen species (ROS), probably through reduced nicotinamide adenine dinucleotide phosphate oxidases, and a late decrease in reduced glutathione content are produced. According to this, antioxidants prevent caspase 8 activation and Bid cleavage, suggesting that ROS production is an important event mediating this process of apoptosis. However, the participation of uncoupling protein-1, -2, and -3 regulating ROS is unclear because their levels remain unchanged upon insulin treatment for 72 h. Our data suggest that the prolonged hyperinsulinemia might cause insulin resistance through the loss of brown adipose tissue.

Identification and conservation of remnant genetic resources of brown trout in relict populations from Western Mediterranean streams

Brown trout is a cold-adapted freshwater species with restricted distribution to headwater streams in rivers of the South European peninsulas, where populations are highly vulnerable because Mediterranean regions are highly sensitive to the global climatic warming. Moreover, these populations are endangered due to the introgressive hybridization with cultured stocks. Individuals from six remnant populations in Western Mediterranean rivers were sequenced for the complete mitochondrial DNA control region and genotyped for 11 nuclear markers. Three different brown trout lineages were present in the studied region. Significant genetic divergence was observed among locations and a strong effect of genetic drift was suggested. An important stocking impact (close to 25%) was detected in the zone. Significant correlations between mitochondrial-based rates of hatchery introgression and water flow variation suggested a higher impact of stocked females in unstable habitats. In spite of hatchery introgression, all populations remained highly differentiated, suggesting that native genetic resources are still abundant. However, climatic predictions indicated that suitable habitats for the species in these rivers will be reduced and hence trout populations are highly endangered and vulnerable. Thus, management policies should take into account these predictions to design upstream refuge areas to protect remnant native trout in the region

Caracterização ácido-base da superfície de espécies mistas da alga Spirulina através de titulação potenciométrica e modelo de distribuição de sítios discretos

Acid base properties of mixed species of the microalgae Spirulina were studied by potentiometric titration in medium of 0.01 and 0.10 mols L-1 NaNO3 at 25.0±0.10 C using modified Gran functions or nonlinear regression techniques for data fitting. The discrete site distribution model was used, permitting the characterization of five classes of ionizable sites in both ionic media. This fact suggests that the chemical heterogeneity of the ionizable sites on the cell surface plays a major role on the acid-base properties of the suspension in comparison to electrostatic effects due to charge-charge interactions. The total of ionizable sites were 1.75±0.10 and 1.86±0.20 mmolsg-1 in ionic media of 0.01 and 0.10 mols L-1 NaNO3, respectively. A major contribution of carboxylic groups was observed with an average 34 and 22% of ionizable sites being titrated with conditional pcKa of 4.0 and 5.4, respectively. The remaining 44% of ionizable sites were divided in three classes with averaged conditional pcKa of 6.9, 8.7 and 10.12, which may be assigned respectively to imidazolic, aminic, and phenolic functionalities.

Estudi de la coautoria de publicacions científiques entre UPC i nou universitats dels Estats Units: Brown University, University of Chicago, Northwestern University, Stanford University, Columbia University, Cornell University, Harvard University, Princeton University i Duke University

S'analitza la coautoria de la UPC amb autors vinculats a institucions acadèmiques dels Estats Units, per totes les àrees temàtiques i sense considerar límits cronològics o documentals.

Influência da temperatura do ar de secagem sobre o teor e a composição química do óleo essencial de Lippia alba (Mill) N. E. Brown

Leaves of Lippia alba were submitted to six different drying treatments, using air at ambient temperature and heated up to 80 °C. The essential oil was extracted by steam distillation and analyzed by GC-MS. For the dried leaves, the oil content was reduced by 12 to 17% when compared with the fresh plant (0.66%). The major oil component was citral, representing 76% for the fresh plant, and varying from 82 to 84% for the dried material. These results showed that L. alba can be submitted to a drying process of up to 80 ºC without degradation and/or loss of the major, [LC1] active component.

4-(hidroximetil)-benzenossulfonato de potássio: metabólito inédito isolado da alga marinha Bostrychia tenella (Rhodomelaceae, ceramiales)

Chemical investigation of the dichloromethane/methanol extract of the marine alga Bostrychia tenella has led to the isolation of two aromatic compounds, the new sulfate metabolite potassium 4-(hydroxymethyl)-benzenosulfonate (1) and the compound 1-methoxyphenethyl alcohol (2), described previously as a synthetic product. Their structures were determined by spectroscopic methods including NMR, MS, IR and by comparison with literature data.

Variabilidade sazonal e biossíntese de terpenóides presentes no óleo essencial de Lippia alba (Mill.) N. E. Brown (Verbenaceae)

The essential oil of the leaves of Lippia alba chemotype linalool-1,8-cineol was extracted by hidrodistillation at different seasons and analyzed by GC/MS. Qualitative and quantitative variations in regard to the period of harvesting have been performed and the results were correlated with meteorological data. The essential oil yield varied from 0.33 to 0.67%. The chemical diversity of the constituents increased throughout the year, being 1,8-cineol and linalool the major components. Possible biosynthetic routes of mono and sesquiterpenoids present in the essential oil are discussed.

Extração e purificação de clorofila a, da alga Spirulina maxima: um experimento para os cursos de química

This work describes a simple and economical experiment for the extraction and purification of chlorophyll a from Spirulina maxima. Extraction and purification of natural compounds can be considered one of the most illustrative experiments that can be performed in Organic Chemistry courses. Particularly, the chromatography of dyes and pigments allows students to have a better comprehension of the chromatography separations. These compounds represent an important class of organic pigments applied in pharmaceutical, cosmetic, detergent compositions, and various other fields and can be extracted from plants and algae. To extract, separate and purify chlorophyll a from associated pigments such as xanthophylls, carotenes, and pheophytins, very costly processes are reported. The present approach is perfectly adequate for use in Chemistry experiments for undergraduate students.

Atividade inseticida de óleos essenciais de Pelargonium graveolens l'Herit e Lippia alba (Mill) N. E. Brown sobre Spodoptera frugiperda (J. E. Smith)

Insecticidal activity of essential oils of Pelargonium graveolens, Lippia alba and compounds geraniol, linalool, 1,8-cineole, limonene, carvone, citral and Azamax® were evaluated against Spodoptera frugiperda. Topical application assay showed essential oil of P. graveolens has acute toxicity against Spodoptera frugiperda larvae (third instar) with LD50 1.13 µg/mg per insect and LD90 2.56 µg/mg per insect. Three essential oils of L. alba also exhibited insecticidal activity with LD50 ranging from 1.20 to 1.56 µg/mg per insect and LD90 from 2.60 to 3.75 µg/mg per insect. Geraniol, linalool, carvone and citral caused significant mortality of 30, 90, 84 and 64% respectively, compared to negative control. The bioinsecticide, Azamax®, caused lower mortality than the compounds of the essential oils.

Determinação de bromo e iodo em alga marinha comestível por ICP-MS após decomposição por combustão iniciada por micro-ondas

A method based on microwave-induced combustion (MIC) was applied for the decomposition of different types of edible seaweed (Nori, Hijiki and Wakame) for subsequent determination of bromine and iodine by inductively coupled plasma mass spectrometry (ICP-MS). Decomposition of 500 mg of each sample was achieved in less than 30 min. A single and diluted solution (150 mmol L-1 (NH4)2CO3) was used for the absorption of both analytes and a reflux step of 5 min was applied to improve analyte recoveries. Accuracy was evaluated using certified reference materials and agreement was between 103 and 108% for both elements.