Spatio-temporal variability in fatty acid trophic biomarkers in stomach contents and muscle of Iberian sardine (<Emphasis Type="Italic">Sardina pilchardus</Emphasis>) and its relationship with spawning

Temporal variation in the fatty acid (FA) composition of stomach contents of Iberian sardines was compared to the relative contribution to dietary carbon made by different prey types for fish from two areas off Portugal. The effect of the FA content of the diet on sardine muscle FA composition was also studied, aiming at (1) analysing if FA biomarkers can be used as a complementary technique for the study of sardine diet and (2) to relate spatial and temporal variations of prey FA content with sardine condition and reproduction. Significant spatial differences in the FA composition of sardine diet occurred with concentrations of n-3 polyunsaturated FA, namely eicosapentaenoic acid [EPA, 20:5n-3] and linolenic acid 18:3n-3, being significantly higher in the diet of sardines from the west coast, whilst the diet of sardines from the south coast was richer in monounsaturated fatty acids (MUFA), namely the carnivory biomarker oleic acid 18:1n-9. These results are in agreement with the higher contribution made by diatoms and dinoflagellates to the diet of sardines off the west coast. Spatial variation in sardine dietary FA was also detected in their muscle composition, specifically for EPA, and the eicosapentaenoic/docosahexaenoic acid and (n-3)/(n-6) ratios, which were higher in sardines from the west coast. No difference in FA composition was detected between sexes, and the seasonal variability in sardine total FA concentration was primarily related to the seasonality of spawning. Sardines accumulate high concentrations of FAs during the resting stage of reproduction when the feeding intensity is similar or lower to that observed during the spawning season. Additionally, sardines show a high selective retention of MUFA and polyunsaturated FA (PUFA) throughout the year except at the beginning of the spawning season, when these FAs are largely invested in the formation of the gonads. Therefore, temporal and regional differences of prey environments are strong enough to be reflected in fish body composition, namely on the accumulation of essential FAs, which can have a strong impact on reproduction success for this species.     

Ranked set sampling allocation models for multiple skewed variables: an application to agricultural data

The mean of a balanced ranked set sample is more efficient than the mean of a simple random sample of equal size and the precision of ranked set sampling may be increased by using an unbalanced allocation when the population distribution is highly skewed. The aim of this paper is to show the practical benefits of the unequal allocation in estimating simultaneously the means of more skewed variables through real data. In particular, the allocation rule suggested in the literature for a single skewed distribution may be easily applied when more than one skewed variable are of interest and an auxiliary variable correlated with them is available. This method can lead to substantial gains in precision for all the study variables with respect to the simple random sampling, and to the balanced ranked set sampling too.     

Assessment of Iodine Direct Titration in Sea Water by Constant Current Potentiometry

Abstract

Dissolved iodine in sea water, about 60 μg L^?1, usually exists in equal proportions as iodate and iodide ions. For their determination several techniques have been proposed (Riley, 1975); among them the volumetric one which depends upon the release of elementary iodine by the addition of acid and iodide ions to the iodate present in the sample. The liberated iodine is titrated with thiosulphate the equivalence point being detected photometrically (Matthews and Riley, 1970; Wong and Brewer, 1974) or it can also be reduced with an excess of thiosulphate and the residual thiosulphate back-titrated with iodate amperometrically (Barkley and Thompson, 1960; Truesdale and Spencer, 1974; Tsonkova and Kulev, 1981). To titrate total iodine in the sample the iodide must be previously oxidized to iodate with bromine (Barkley and Thompson, 1960; Tsonkova and Kulev, 1981) or permanganate (Schnepfe, 1972). In the amperometric procedure Truesdale and Spencer (1974) observed that platinum electrodes often do not respond reliably.     

Isolation of single circulating trophoblasts from maternal circulation for noninvasive fetal copy number variant profiling

Objective

To develop a multi-step workflow for the isolation of circulating extravillous trophoblasts (cEVTs) by describing the key steps enabling a semi-automated process, including a proprietary algorithm for fetal cell origin genetic confirmation and copy number variant (CNV) detection.

Methods

Determination of the limit of detection (LoD) for submicroscopic CNV was performed by serial experiments with genomic DNA and single cells from Coriell cell line biobank with known imbalances of different sizes. A pregnancy population of 372 women was prospectively enrolled and blindly analyzed to evaluate the current workflow.

Results

An LoD of 800 Kb was demonstrated with Coriell cell lines. This level of resolution was confirmed in the clinical cohort with the identification of a pathogenic CNV of 800 Kb, also detected by chromosomal microarray. The mean number of recovered cEVTs was 3.5 cells per sample with a significant reverse linear trend between gestational age and cEVT recovery rate and number of recovered cEVTs. In twin pregnanices, evaluation of zygosity, fetal sex and copy number profiling was performed in each individual cell.

Conclusion

Our semi-automated methodology for the isolation and single-cell analysis of cEVTS supports the feasibility of a cell-based noninvasive prenatal test for fetal genomic profiling.     

Study concerning the recovery of zinc and manganese from spent batteries by hydrometallurgical processes

Used batteries contain numerous metals in high concentrations and if not disposed of with proper care, they can negatively affect our environment. These metals represent 83% of all spent batteries and therefore it is important to recover metals such as Zn and Mn, and reuse them for the production of new batteries. The recovery of Zn and Mn from used batteries, in particular from Zn–C and alkaline ones has been researched using hydrometallurgical methods. After comminution and classification of elemental components, the electrode paste resulting from these processes was treated by chemical leaching. Prior to the leaching process the electrode paste has been subjected to two washing steps, in order to remove the potassium, which is an inconvenient element in this type of processes. To simultaneously extract Zn and Mn from this paste, the leaching method in alkaline medium (NaOH solution) and acid medium (sulphuric acid solution) was used. Also, to determine the efficiency of extraction of Zn and Mn from used batteries, the following variables were studied: reagents concentration, S/L ratio, temperature, time. The best results for extraction yield of Zn and Mn were obtained under acid leaching conditions (2 M H₂SO₄, 1 h, 80 °C).     

Evaluation of the measurement uncertainty in automated long-term sampling of PCDD/PCDFs

Since the publication of the first version of European standard EN-1948 in 1996, long-term sampling equipment has been improved to a high standard for the sampling and analysis of polychlorodibenzo-p-dioxin (PCDD)/polychlorodibenzofuran (PCDF) emissions from industrial sources. The current automated PCDD/PCDF sampling systems enable to extend the measurement time from 6–8 h to 15–30 days in order to have data values better representative of the real pollutant emission of the plant in the long period. EN-1948:2006 is still the European technical reference standard for the determination of PCDD/PCDF from stationary source emissions. In this paper, a methodology to estimate the measurement uncertainty of long-term automated sampling is presented. The methodology has been tested on a set of high concentration sampling data resulting from a specific experience; it is proposed with the intent that it is to be applied on further similar studies and generalized. A comparison between short-term sampling data resulting from manual and automated parallel measurements has been considered also in order to verify the feasibility and usefulness of automated systems and to establish correlations between results of the two methods to use a manual method for calibration of automatic long-term one. The uncertainty components of the manual method are analyzed, following the requirements of EN-1948-3:2006, allowing to have a preliminary evaluation of the corresponding uncertainty components of the automated system. Then, a comparison between experimental data coming from parallel sampling campaigns carried out in short- and long-term sampling periods is realized. Long-term sampling is more reliable to monitor PCDD/PCDF emissions than occasional short-term sampling. Automated sampling systems can assure very useful emission data both in short and long sampling periods. Despite this, due to the different application of the long-term sampling systems, the automated results could not be directly compared with manual results, not even in terms of measurement uncertainty. This investigation focuses on both uncertainty and repeatability of the automated sampling method. The standard 20988, developed by Internarional Organization of Standardization (ISO) can be used to estimate the measurement uncertainty. The results confirm that the uncertainties of manual and automated methods are comparable. At the same time, it is not appropriate to consider the manual method as a reference for the evaluation of the uncertainty of the automated sampling system, due to the high variability of both systems.     

Assessment of Iodine Direct Titration in Sea Water by Constant Current Potentiometry

Dissolved iodine in sea water, about 60 μg L^-1, usually exists in equal proportions as iodate and iodide ions. For their determination several techniques have been proposed (Riley, 1975); among them the volumetric one which depends upon the release of elementary iodine by the addition of acid and iodide ions to the iodate present in the sample. The liberated iodine is titrated with thiosulphate the equivalence point being detected photometrically (Matthews and Riley, 1970; Wong and Brewer, 1974) or it can also be reduced with an excess of thiosulphate and the residual thiosulphate back-titrated with iodate amperometrically (Barkley and Thompson, 1960; Truesdale and Spencer, 1974; Tsonkova and Kulev, 1981). To titrate total iodine in the sample the iodide must be previously oxidized to iodate with bromine (Barkley and Thompson, 1960; Tsonkova and Kulev, 1981) or permanganate (Schnepfe, 1972). In the amperometric procedure Truesdale and Spencer (1974) observed that platinum electrodes often do not respond reliably.     

Genotoxicity of gold nanoparticles functionalized with indolicidin towards Saccharomyces cerevisiae

The toxic effects of gold nanoparticles surface-functionalized with the antimicrobial peptide indolicidin(Au NPs-indolicidin) towards the yeast Saccharomyces cerevisiae, one of the major eukaryotic model organisms, have been evaluated. Growth and survival,genotoxicity, as measured by comet assay, and expression of the YCA1, an apoptosis indicating gene, following 72 hr exposure of yeast to Au NPs-indolicidin, and to Au NPs and indolicidin alone have been examined. The gold nanoparticles exerted toxicity with DNA damage, accompanied by reactive oxygen species production(ROS), but they do not inhibit yeast growth and viability. Genotoxicity was less pronounced for surface-functionalized nanoparticles, showing that S. cerevisiae is quite resistant to the complex Au NPs-indolicidin.A progressive reduction of the genotoxic effect was observed along 72 hr exposure,presumably due to the activation of DNA repair mechanisms. These findings suggest the occurrence of a physiological protective response of S. cerevisiae towards nanoparticles,thereby providing useful information to the assessment of the environmental impact of metal nanoparticles.