Enriching rotifers with “premium” microalgae. Isochrysis aff. galbana clone T-ISO

The effect of semi-continuous culture on the nutritional value of microalgae was tested in the rotifer Brachionus plicatilis in short-term enrichment experiments. Isochrysis aff. galbana clone T-ISO was cultured semi-continuously with renewal rates from 10 to 50% of the volume of the culture per day and used to feed the rotifers. After 24 h, dramatic differences in dry weight and protein, lipid and carbohydrate contents were observed in the rotifers depending on the renewal rate applied to the microalgal culture. Rotifers fed T-ISO cultured with low renewal rates showed low dry weight and organic content, whereas rotifers fed microalgae from nutrient-sufficient, high renewal rate cultures showed higher dry weight and increases up to 60% in protein, 35% in lipid and 100% in carbohydrate contents. Feed conversion rate (FCR) and organic FCR decreased with increasing renewal rates, indicating a more efficient assimilation of the microalgal biomass obtained at high growth rates. The fatty acid profile of rotifers reflected that of T-ISO, with maximum content of polyunsaturated fatty acids (PUFAs), n-3 fatty acids and docosahexaenoic acid (DHA) being found in the rotifers fed microalgae from the renewal rate of 40%. Results demonstrate that the biochemical composition of B. plicatilis is strongly modified through the use of semi-continuous cultures of microalgae in short-term enrichment processes. This technique provides an excellent tool to improve the nutritional value of the live feed used in fish larvae cultures.     

S‐locus diversity and cross‐compatibility of wild Prunus avium for timber breeding

Prunus avium is primarily cultivated for its fruit, sweet cherries. However, it is also used to produce high‐quality timber. In a P. avium seed orchard, gametophytic self‐incompatibility is a restriction for free pollen flow and should be considered when establishing basic forest materials. In this study, S‐locus diversity and cross‐incompatibility of wild cherry individuals in clonal banks established for breeding for timber production were investigated. Wild cherry trees (140) with outstanding forest growth habit, collected in northern Spain, grafted and planted in two clonal banks, were genotyped at the S‐locus. The self‐incompatibility S‐locus genes, S‐RNase and SFB, were analysed by PCR. Twenty‐two S‐haplotypes, resulting in 72 different S‐genotypes, were identified. The genotypes were grouped into 33 incompatibility groups and 39 unique genotypes. This initial S‐locus analysis revealed large genetic diversity of wild cherry trees from the Spanish northern deciduous forest, and provides useful information for seed orchard design. Wild P. avium displays significantly more genetic diversity than what is detected in local cultivars, revealing a narrowing of genetic diversity during local domestication.     

<Emphasis Type="Italic">Lupinus albus</Emphasis> Conglutin Gamma Modifies the Gene Expressions of Enzymes Involved in Glucose Hepatic Production <Emphasis Type="Italic">In Vivo</Emphasis>

Lupinus albus seeds contain conglutin gamma (Cγ) protein, which exerts a hypoglycemic effect and positively modifies proteins involved in glucose homeostasis. Cγ could potentially be used to manage patients with impaired glucose metabolism, but there remains a need to evaluate its effects on hepatic glucose production. The present study aimed to analyze G6pc, Fbp1, and Pck1 gene expressions in two experimental animal models of impaired glucose metabolism. We also evaluated hepatic and renal tissue integrity following Cγ treatment. To generate an insulin resistance model, male Wistar rats were provided 30% sucrose solution ad libitum for 20 weeks. To generate a type 2 diabetes model (STZ), five-day-old rats were intraperitoneally injected with streptozotocin (150 mg/kg). Each animal model was randomized into three subgroups that received the following oral treatments daily for one week: 0.9% w/v NaCl (vehicle; IR-Ctrl and STZ-Ctrl); metformin 300 mg/kg (IR-Met and STZ-Met); and Cγ 150 mg/kg (IR-Cγ and STZ-Cγ). Biochemical parameters were assessed pre- and post-treatment using colorimetric or enzymatic methods. We also performed histological analysis of hepatic and renal tissue. G6pc, Fbp1, and Pck1 gene expressions were quantified using real-time PCR. No histological changes were observed in any group. Post-treatment G6pc gene expression was decreased in the IR-Cγ and STZ-Cγ groups. Post-treatment Fbp1 and Pck1 gene expressions were reduced in the IR-Cγ group but increased in STZ-Cγ animals. Overall, these findings suggest that Cγ is involved in reducing hepatic glucose production, mainly through G6pc inhibition in impaired glucose metabolism disorders.     

Supercritical fluid extraction of pesticides from a table-ready food composite of plant origin (gazpacho)

Supercritical fluid carbon dioxide extraction (SFE) has been evaluated for the extraction of 17 organohalogen and organophosphate pesticides in gazpacho (a table-ready food composite containing crude vegetables, white bread, vegetable oil, water, and other minor components) using anhydrous magnesium sulfate as drying agent. The effects of different parameters, such as fat content in gazpacho composites, magnesium sulfate/gazpacho ratio, supercritical fluid volume, pressure, temperature, and static modifier additions, on SFE recoveries from spiked gazpacho samples have been studied. Analyses were performed by gas chromatography (GC) with flame photometric (FPD), electron capture (ECD), and mass spectrometry (MSD) detectors. In most experiments, recoveries obtained for the nonpolar organohalogen pesticides were lower than those obtained for the most polar organophosphate pesticides, but overall pesticide recoveries determined by using the optimal SFE conditions indicate that SFE could be used to determine pesticide residue levels in gazpacho.     

Direct analysis of pesticide residues in olive oil by on-line reversed phase liquid chromatography-gas chromatography using an automated through oven transfer adsorption desorption (TOTAD) interface

A fully automated on-line reversed phase liquid chromatography-gas chromatography system is described. The system uses a prototype of the automated through oven transfer adsorption desorption interface. The system is demonstrated by presenting a new rapid method for the determination of pesticide residue in olive oil, which is injected directly with no sample pretreatment step other than filtration. Methanol:water is used as the eluent in the LC preseparation step, while the LC fraction containing the pesticide is automatically transferred to the gas chromatograph. Detection limits of pesticides varied from 0.18 to 0.44 mg/L when a flame ionization detector was used. As an example, relative standard deviation and linear calibration are presented for terbutryne.     

Phenolic profile of quince fruit (Cydonia oblonga Miller) (pulp and peel)

Qualitative and quantitative analyses of phenolic compounds were carried out on quince fruit samples from seven different geographical origins in Portugal. For each origin, both pulp and peel were analyzed by reversed-phase HPLC-DAD and HPLC-DAD/MS.The results revealed differences between the phenolic profiles of pulps and peels in all studied cases. The pulps contained mainly caffeoylquinic acids (3-, 4-, and 5-O-caffeoylquinic acids and 3,5-dicaffeoylquinic acid) and one quercetin glycoside, rutin (in low amount). The peels presented the same caffeoylquinic acids and several flavonol glycosides: quercetin 3-galactoside, kaempferol 3-glucoside, kaempferol 3-rutinoside, and several unidentified compounds (probably kaempferol glycoside and quercetin and kaempferol glycosides acylated with p-coumaric acid). The highest content of phenolics was found in peels.     

Use of nootkatone as a senescence indicator for Rouge La Toma Cv. grapefruit (Citrus paradisi Macf.)

The objective of this research was to study the usefulness of nootkatone as a senescence indicator for Rouge La Toma cv. grapefruit (Citrus paradisi Macf.), simulating different treatments that included the normal postharvest handling of citrus fruits: temperature conditioning, cold storage, shipment periods to overseas markets such as Japan and the U.S., marketing conditions, and storage at nonchilling temperature (control treatments). The highest nootkatone levels, determined by GLC-MS analyses, were detected in fruits subjected to control treatments. No significant differences were observed in nootkatone levels between treatments either with or without temperature conditioning prior to the start of the cold storage. Levels of nootkatone increased throughout time for all assayed treatments. The linear regressions of nootkatone levels showed correlation coefficients of 0.80 and 0.83 with storage time (29 and 42 days, respectively). Therefore, nootkatone appears to be a good indicator of senescence for Rouge La Toma grapefruit.     

Time-resolved spectral studies of blue-green fluorescence of artichoke (Cynara cardunculus L. Var. Scolymus) leaves: identification of chlorogenic acid as one of the major fluorophores and age-mediated changes

Synchrotron radiation and the time-correlated single-photon counting technique were used to investigate the spectral and time-resolved characteristics of blue-green fluorescence (BGF) of artichoke leaves. Leaves emitted BGF under ultraviolet (UV) excitation; the abaxial side was much more fluorescent than the adaxial side, and in both cases, the youngest leaves were much more fluorescent than the oldest ones. The BGF of artichoke leaves was dominated by the presence of hydroxycinnamic acids. A decrease in the percentage of BGF attributable to the very short kinetic component (from 42 to 20%), in the shape of the BGF excitation spectra, and chlorogenic acid concentrations indicate that there is a loss of hydroxycinnamic acid with leaf age. Studies on excitation, emission, and synchronized fluorescence spectra of leaves and trichomes and chlorogenic acid contents indicate that chlorogenic acid is one of the main blue-green fluorophores in artichoke leaves. Results of the present study indicate that 20-42% (i.e., the very short kinetic component) of the overall BGF is emitted by chlorogenic acid. Time-resolved BGF measurements could be a means to extract information on chlorogenic acid fluorescence from the overall leaf BGF.     

Occurrence of naturally acetylated lignin units

This work examines the occurrence of native acetylated lignin in a large set of vascular plants, including both angiosperms and gymnosperms, by a modification of the so-called Derivatization Followed by Reductive Cleavage (DFRC) method. Acetylated lignin units were found in the milled wood lignins of all angiosperms selected for this study, including mono- and eudicotyledons, but were absent in the gymnosperms analyzed. In some plants (e.g., abaca, sisal, kenaf, or hornbeam), lignin acetylation occurred at a very high extent, exceeding 45% of the uncondensed (alkyl-aryl ether linked) syringyl lignin units. Acetylation was observed exclusively at the gamma-carbon of the lignin side chain and predominantly on syringyl units, although a predominance of acetylated guaiacyl over syringyl units was observed in some plants. In all cases, acetylation appears to occur at the monomer stage, and sinapyl and coniferyl acetates seem to behave as real lignin monomers participating in lignification.