Optimization of the extraction of antioxidants from Dunaliella salina microalga by pressurized liquids

In this work, extraction of antioxidant compounds from Dunaliella salina microalga is optimized by combining pressurized liquid extraction (PLE) and experimental design (three-level factorial design) with three different solvents (hexane, ethanol, and water). Two main factors were considered, the extraction temperature (40, 100, and 160 degrees C) and the extraction time (5, 17.5, and 30 min). As response variables, the extraction yield (percent dry weight/initial weight) and the antioxidant activity of the extracts (determined using the TEAC method) were used. The parameters of the model were estimated by multiple linear regression. Results showed that the extraction temperature was the factor having the strongest influence (positive) on the two response variables. The best yields were obtained with ethanol at the higher extraction temperature and time tested. Besides, although hexane extracts provided the best antioxidant activity, ethanol extracts were also very active. The chemical characterization of ethanol extracts was carried out using HPLC-DAD, and attempts have been made to correlate their chemical composition with the antioxidant activity measured. Results pointed out that the extracts contained, besides all-trans-beta-carotene and isomers, several different minor carotenoids that seemed to make a contribution to the antioxidant activity of the extracts.     

Antiproliferative Activity of Fucan Nanogel

Sulfated fucans comprise families of polydisperse natural polysaccharides based on sulfated L-fucose. Our aim was to investigate whether fucan nanogel induces cell-specific responses. To that end, a non toxic fucan extracted from Spatoglossum schröederi was chemically modified by grafting hexadecylamine to the polymer hydrophilic backbone. The resulting modified material (SNFuc) formed nanosized particles. The degree of substitution with hydrophobic chains was close to 100%, as estimated by elemental analysis. SNFfuc in aqueous media had a mean diameter of 123 nm and zeta potential of −38.3 ± 0.74 mV, as measured by dynamic light scattering. Nanoparticles conserved their size for up to 70 days. SNFuc cytotoxicity was determined using the MTT assay after culturing different cell lines for 24 h. Tumor-cell (HepG2, 786, H-S5) proliferation was inhibited by 2.0%–43.7% at nanogel concentrations of 0.05–0.5 mg/mL and rabbit aorta endothelial cells (RAEC) non-tumor cell line proliferation displayed inhibition of 8.0%–22.0%. On the other hand, nanogel improved Chinese hamster ovary (CHO) and monocyte macrophage cell (RAW) non-tumor cell line proliferation in the same concentration range. The antiproliferative effect against tumor cells was also confirmed using the BrdU test. Flow cytometric analysis revealed that the fucan nanogel inhibited 786 cell proliferation through caspase and caspase-independent mechanisms. In addition, SNFuc blocks 786 cell passages in the S and G2-M phases of the cell cycle.     

Does otolith macrostructure record environmental or biological events? The case of black hake (Merluccius polli and Merluccius senegalensis)

Fish age determination using otoliths requires a prior understanding of growth mark deposition patterns (translucent rings, TR) as well as their connection with internal or external events experienced by the fish. This study analysed the macrostructural seasonal ring deposition pattern observed in transversal sections of black hake otoliths. A total of 793 black hake otoliths were collected in autumn and spring 2007 from research and commercial surveys carried out in continental and shelf waters off Mauritania. Most of the Merluccius polli otoliths presented narrow and wide translucent rings (NTR and WTR, respectively) regardless of fish size, whereas Merluccius senegalensis otoliths only showed NTR. This seemed to be a sign of ontogenetic discrepancy between the two black hake species, whose otoliths confirmed the existence of significant differences in their growth patterns.The frequency distributions of the number of TR counted along the ventral radius (VR) of the otolith from the nucleus (birth date) and from the ventral edge (death date) were analysed to ascertain whether a specific endogenous event (Hypothesis A) or a precise environmental event (Hypothesis B) could restrain growth leading to the formation of TR. The general TR frequency distribution pattern was somewhat similar for both hake species, which showed marked TR at comparable distances. Within each species, TR frequency distributions of their distances from the nucleus along the otolith VR were quite similar between sexes, seasons, and fish sizes. Our results support the idea of a coincident biological event, such as first maturity, slowing down the growth process and thus provoking the formation of TR in otoliths of both species.This study also experience difficulty using the complex and highly variable macrostructural pattern of black hake otoliths to establish age interpretation criteria for these two species.     

Environmental effects of a marine fish farm of gilthead seabream (Sparus aurata) in the NW Mediterranean Sea on water column and sediment

This study examined the effects of organic enrichment on water column, sediments and macrofauna caused by a fish farm in the Mediterranean Sea. Samples were collected on four sampling campaigns over a one‐year cycle. Significant differences were found in the water column in dissolved oxygen, dissolved inorganic nitrogen, phosphate and total phosphorus concentrations between the fish farm and the control. The increase in the dissolved inorganic nitrogen and phosphate concentrations at the fish farm modified the stoichiometric ratios between nutrients, with silicate acting as limiting nutrient at the fish farm 11% more than at the control. Nevertheless, chlorophyll a concentration in the water column was higher at the control station, probably due to the fouling of the underwater fish farm structures. Significant differences were found in sediment concentrations of organic matter, total phosphorus and redox potential between the fish farm and the control. The Canonical Correlation Analysis indicated that organic matter, total phosphorus, redox potential and% of gravels accounted for 68.9% of the total variance in the species data. Changes were observed in macrofauna, with a decrease in number of species and up to a nine‐fold increase in abundance with respect to the control.     

A comparison of indexes to estimate corn S uptake and S mineralization in the field

The development of simple predictors of sulfur (S) mineralization and its correlation with field-derived data may help improving corn S availability diagnosis. The objectives of this study were (1) to compare methods to estimate soil S mineralization, (2) to develop a model to predict soil S mineralization from S mineralization indexes and edaphic variables, and (3) to predict field-grown corn S uptake (S_uptake) and apparent S mineralization (S_min-app) from different S mineralization indexes and edaphic-climatic variables. We evaluated 26 experimental sites where we measured edaphic variables as soil organic C (SOC), organic C in the particulate fraction (C-PF), S mineralization potential (S_min-10wk), S mineralized during a short-term (7 days) aerobic incubation + initial inorganic S (S_min-7d?+?S_inorg), and N mineralized during a short-term (7 days) anaerobic incubation (N_an). Additionally, 18 field experiments were carried out to quantify S_uptake and S_min-app. The C-PF, S_min-7d?+?S_inorg, N_an, and SOC were variables significantly correlated with S_min-10wk (r?=?0.89, 0.89, 0.88, and 0.85, respectively). We developed a simple model to predict S_min-10wk from selected edaphic variables (S_min-10wk?=?0.038*N_an?+?0.106*SOC?+?0.74; R_a² =?0.87). The S_min-10wk, C-PF, and S_min-7d?+?S_inorg showed a liner-plateau association with S_uptake (R²?=?0.73, 0.53, and 0.48, respectively). We modified the method to estimate S_min-app to account for S losses (S_{min-app (modified)}) and developed a model to predict S_{min-app (modified)} from C-PF (S_{min-app (modified)}?=?4.65*C-PF?+?9.86; R²?=?0.62) or S_min-10wk (S_{min-app (modified)}?=?3.0*S_min-10wk?+?7.4; R²?=?0.54). Our results demonstrate that S mineralization indexes can be used to predict corn S availability under field conditions.     

Does the persistence of sweet chestnut depend on cultural inputs? Regeneration,recruitment, and mortality in <Emphasis Type="Italic">Quercus</Emphasis>- and <Emphasis Type="Italic">Castanea</Emphasis>-dominated forests

Key message

Quercus secondary forests show a gradual transition toward mixed forests, with sweet chestnut ( Castanea sativa ) becoming increasingly abundant in the western Spanish Central System. Additionally, in chestnut-dominated stands, it shows a certain resistance to competitive displacement by Quercus pyrenaica . Our results partially refute the traditional view that C. sativa is unable to recruit in the absence of cultural inputs.

Context

Sweet chestnut, Castanea sativa, is a component of European broadleaf forests and is one of the most managed trees. Due to a reduction in cultural inputs, chestnut-dominated stands tend to be invaded by other species, and it is unclear how chestnut is able to persist in natural mixed forests.

Aims

Our work aimed to identity the main factors that limit the establishment of C. sativa and to analyze the recruitment and mortality processes of C. sativa trees.

Methods

The age, growth ring patterns, regeneration density, and the spatial structure of trees and saplings in 11 plots in the Spanish Central System were analyzed.

Results

Chestnut seedling density increased with C. sativa basal area, but transition toward the sapling stage appeared limited owing to light availability. In Quercus pyrenaica secondary forests, sparse canopies did not constrain chestnut regeneration, and in old chestnut stands, C. sativa showed a certain resistance to competitive displacement. By contrast, mixed young coppices showed a high mortality, most likely due to competition with other vigorous resprouters.

Conclusion

Quercus secondary forests showed a gradual transition toward mixed forests with sweet chestnut becoming increasingly more abundant. In old stands, C. sativa is likely to persist under a gap-phase mode of regeneration. Our results partially refute the traditional view that C. sativa is unable to recruit in the absence of cultural inputs.

     

Pathogenicity of plant and soil isolates of <Emphasis Type="Italic">Phytophthora parasitica</Emphasis> on tomato and pepper

Crown and root rot of tomato and sweet pepper can be caused by Phytophthora parasitica. In this work, 23 P. parasitica isolates from diseased pepper or tomato plants as well as 54 isolates from 23 monocrop tomato soils (from Spain and Chile) and one from a pepper soil were studied for their host–pathogen response. Results show significant host specificity for the isolates from tomato plants and tomato soils (63 of 64 isolates were unable to cause disease in pepper). None of the pepper plant/soil isolates showed pathogenicity on tomato, and only four of 14 reproduced their pathogenicity on pepper. Only one tomato isolate was pathogenic to both Solanaceae species. Two different inoculation protocols were evaluated (substrate irrigation and stem cutting). All isolates which expressed pathogenicity when stem inoculated also did it when root inoculated, but not vice-versa. Therefore, the recommended test protocol for tomato and pepper breeding programmes is that based on root inoculation by irrigation.