Preliminary chemometric study on the use of honey as an environmental marker in Galicia (northwestern Spain)

Thirteen metal elements were determined in 40 honey samples from Galicia with different environmental origins: rural, urban, and industrial areas. The data set of the honey metallic profiles was studied with a double purpose: first, to make a preliminary evaluation of honey as an environmental indicator in Galicia with the aim of monitoring pollution and, second, to compare the different capabilities of diverse pattern recognition prediction procedures for modeling the environmental surrounding of the hive. A certain level of similarity for urban and industrial samples was obtained using principal component analysis and cluster analysis, whereas significant differences for urban and industrial honeys were found in relation to rural honey samples. Different classification rules to associate metal content of honeys with their environmental surrounding were obtained by chemometric pattern recognition procedures. In general, the classification methods developed by neural networks provided better results than the traditional pattern recognition procedures. The metal profiles of honey seem to provide sufficient information to enable categorization criteria for classifying samples according to their environmental surrounding. Thus, honey could be a potential pollution indicator for the Galician area.     

Determination of dexamethasone in bovine liver by chemiluminescence high-performance liquid chromatography.

A new method for the determination of dexamethasone (9alpha-fluoro-11beta,17alpha,21-trihydroxy-16alpha -methylpregna-1, 4-diene-3,20-dione) in bovine liver was developed. This new liquid-liquid extraction method comprises the addition of sodium hydroxide to the tissue sample followed by extraction with ethyl acetate. After centrifugation, the extract is evaporated to dryness and the residue dissolved in acetonitrile. The cleaning of the fat is performed with n-hexane, and the acetonitrile layer is evaporated. Analysis of the extracts is performed using high-performance liquid chromatography with chemiluminescence detection employing luminol as CL reagent. A series of recovery curves performed at spiking levels of 50, 30, 10, 5, and 2.5 ppb show that at least 80% of DEX can be recovered from liver and that the chemiluminescence detection yields satisfactory results with respect to sensitivity (LOD 0.2 ppb), reproducibility (CV% 10.7) and repeatability (CV% 6.2-8.9).     

The first spontaneous spawning of European hake Merluccius merluccius L.: characteristics of eggs and early larval stages

For the first time, a spontaneous spawning of hake was recorded in Spain in April 2009. The spawn was obtained from broodstock kept in captivity for two years at the facilities of the Spanish Institute of Oceanography in Vigo (NW Spain). Eggs were transparent, spherical and had an average diameter of 1.067 ± 0.024 mm; yolk occupied the majority of egg volume. The oil droplet had a diameter of 0.27 ± 0.03 mm. The incubation period of the eggs lasted for 4 days at 14°C and the duration from hatching to the total absorption of the yolk sac was between 5–7 days after hatching, at the same temperature. Newly hatched larvae had an average total length of 3.20 ± 0.13 mm and began feeding 6 days after hatching; a daily growth rate of 0.158 mm day‐1 was observed from hatching to yolk sac consumption. This paper describes the daily evolution of biometric and morphological characteristics of the different stages of embryos and larvae of European hake up to the age of 19 days.     

Soil biology and warming play a key role in the release of ‘old C’ from organic soils

A radiocarbon approach was used to investigate the roles of temperature and soil fauna activity in the turnover of ‘old’ non-labile carbon in a peatland ecosystem. We investigated the impacts of enchytraeids on carbon turnover in two different soil layers, with different incorporation of the ‘bomb’ peak, when incubated at two different temperatures. Results showed that, in agreement with previous studies, warmer temperatures promoted reproduction rates of enchytraeids, with the top layer supporting higher animal densities and biomass. With independence of the animal treatment, soil respiration in the top 5 cm was four times higher than in the deeper layer suggesting that decomposition was greater in the upper layer, with the response being greater at the highest temperature treatment. Furthermore, independent of temperature, the presence of enchytraeids in the top layer significantly enhanced the release of non-labile C as DOC. Similarly, at the bottom layer, ‘older’ C sources were mobilised in response to warming and a greater amount of pre-bomb carbon was released into the soil solution at 20 °C when the worms were present. A strong positive link between the ages of the C assimilated by the animals and released through mineralization suggests an important role of soil biology in the mobilisation of the older C pools in soils and should be taken into account in developing global C models to predict the response of soil C dynamics to climate change.     

Morphological development and forage quality changes in crimson clover (Trifolium incarnatum L.)

The development of efficient forage management and utilization is facilitated by a knowledge of the changes in nutritive value and patterns of dry matter (DM) distribution within plants. The objectives of this experiment were to determine the patterns of DM accumulation and the change in nutritive value during maturation of crimson clover ( Trifolium incarnatum L.), a cool-season annual forage legume adapted to mild winter areas and acid soils. Crimson clover was established on a Dystric Eurocrept soil with pH 5·4 in Galicia, north-western Spain, during three consecutive years (1990–92). Shoot samples were taken at approximately biweekly intervals in the spring and summer after autumn sowing, and were separated into stems, leaves and seed pods. DM accumulation, crude protein (CP) and acid-detergent fibre (ADF) concentrations and in vitro organic matter digestibility (IVOMD) were determined for each plant part at each stage. Average annual DM production at initial flowering was 4·2 t ha^–1, whereas the CP and ADF concentrations and IVOMD values were 134 g kg^–1 DM, 302 g kg^–1 DM and 0·720 respectively. The nutritive value declined with age, coinciding with a reduction in leaf:stem ratio (from 0·53 at the vegetative stage to 0·13 at the ripe seed stage). This decline was more rapid than that reported for other legumes, possibly because of the low leaf:stem ratio of crimson clover. However, as it is used in the Atlantic coastal areas of north-western Spain, where it is harvested around initial flowering, it can provide forage of good quality.     

Chromosome alignment and segregation regulated by ubiquitination of survivin

Proper chromosome segregation requires the attachment of sister kinetochores to microtubules from opposite spindle poles to form bi-oriented chromosomes on the metaphase spindle. The chromosome passenger complex containing Survivin and the kinase Aurora B regulates this process from the centromeres. We report that a de-ubiquitinating enzyme, hFAM, regulates chromosome alignment and segregation by controlling both the dynamic association of Survivin with centromeres and the proper targeting of Survivin and Aurora B to centromeres. Survivin is ubiquitinated in mitosis through both Lys(48) and Lys(63) ubiquitin linkages. Lys(63) de-ubiquitination mediated by hFAM is required for the dissociation of Survivin from centromeres, whereas Lys(63) ubiquitination mediated by the ubiquitin binding protein Ufd1 is required for the association of Survivin with centromeres. Thus, ubiquitinaton regulates dynamic protein-protein interactions and chromosome segregation independently of protein degradation.     

Nitrate improves growth in salt-stressed citrus seedlings through effects on photosynthetic activity and chloride accumulation

We analyzed the effects of nitrate availability on growth of Navelina (Citrus sinensis (L.) Osbeck) scions grafted on three citrus rootstocks differing in salt tolerance: Carrizo citrange (Citrus sinensis (L.) Osbeck x Poncirus trifoliata (L.) Raf.), Citrus macrophylla Wester and Cleopatra mandarin (Citrus reshni Hort. ex Tanaka). Salt stress reduced total plant biomass by 27-38%, whereas potassium nitrate supplementation partially counteracted this effect by increasing dry matter and new leaf area. Salinized Carrizo citrange had the greatest response to nitrate supplementation, whereas the effects on salinized Cleopatra mandarin and C. macrophylla were less apparent. Nitrogen and chlorophyll contents and photosynthetic activity also increased in leaves of the nitrate-supplemented salinized plants. In salinized plants, nitrate supplementation reduced leaf abscission, stimulated photosynthetic activity and increased growth of new leaves. The nitrate treatment did not modify chloride concentration in leaves, but it reduced chloride concentrations in Carrizo and Macrophylla roots. Therefore, in both rootstocks, chloride content was similar in mature leaves, higher in immature leaves and lower in roots of the nitrate-supplemented salinized plants compared with salinized plants unsupplemented with nitrate. We suggest that the nitrate-induced stimulation of growth reduced chloride concentration in roots through the reallocation of chloride to new leaves.