Optimization of large-volume injection for the determination of polychlorinated biphenyls in children's fast-food menus by low-resolution mass spectrometry

This study includes the determination of five indicator polychlorinated biphenyls (PCBs) (52, 101, 153, 138, and 180), six non-ortho PCBs (35, 80, 81, 77, 126, and 169), and two mono-ortho PCBs (28 and 118) in fast food for children. A freeze-dried sample of 10 g is extracted by using pressurized n-hexane in two 5 min cycles at 120 degrees C and 100 mbar. Fatty extracts were cleaned up by means of acetonitrile/n-hexane partitioning and gel-permeation chromatography. The fractionation of non-ortho, mono-ortho, and indicator PCBs was made on graphitized carbon solid-phase extraction cartridges by using n-hexane, n-hexane/toluene (99:1, v/v), and toluene as elution solvents. Gas chromatography coupled to tandem mass spectrometry and large-volume injections with a programmed-temperature vaporizer (PTV-LV) were used to increase sensitivity and selectivity of the PCB determination. The PTV-LV injection settings, that is, vaporizing temperature, vaporizing time, and purge flow, were optimized by using a central composite design. A 15-40 times increased sensitivity was reached as compared with that obtained with the conventional 1 microL splitless injection. The limits of detection achieved were between 0.3 and 1.2 pg/g, and repeatability data, as relative standard deviation varied, ranged from 2 to 9% for the 0.05 ng/mL PCB level.     

Application of comprehensive two-dimensional liquid chromatography to elucidate the native carotenoid composition in red orange essential oil

In the present work, the ability of a LC x LC-DAD/APCI-MS method developed at this laboratory to identify the native composition of carotenoid in an extremely complex matrix such as red orange essential oil was demonstrated. To carry out this task, two independent and orthogonal separation mechanisms were coupled through a 10-port switching valve that simultaneously collected the eluent from a microbore cyano column used as the first dimension in normal phase mode and injected it to a conventional reversed phase monolithic C(18) column in the second dimension separation. By using this novel analytical technique together with the use of DAD and APCI-MS detectors it was possible to identify in the sample, without the need of any pretreatment, 40 different carotenoids. Among them, 16 carotenoid monoesters were identified, mainly beta-cryptoxanthin palmitate (C(16:0)), myristate (C(14:0)), and laureate (C(12:0)) as well as several lutein, violaxanthin, antheraxanthin, and luteoxanthin monoesters. Moreover, 21 carotenoid diesters composed by several antheraxanthin, luteoxanthin, violaxanthin, and auroxanthin diesters were found in the native carotenoid composition of the orange oil. The main carotenoid diesters were the laureate palmitate (C(12:0), C(16:0)), myristate palmitate (C(14:0), C(16:0)), and dipalmitate (C(16:0), C(16:0)) diesters, although other diesters were also identified. Besides, two different free carotenes, zeta-carotene and phytofluene, and a xanthophyll, lutein, were also determined. To the authors' knowledge, this is the first time that carotenoid diesters are described and identified in orange essential oil. Likewise, it has been demonstrated that the LC x LC approach proposed in this study is capable of coping with the direct analysis and identification of a complex natural source of carotenoids such as the orange.     

Diversity,occurrence, and life characteristics of natural entomopathogenic nematode populations from La Rioja (Northern Spain) under different agricultural management and their relationships with soil factors

Natural entomopathogenic nematode (EPN) populations were surveyed in La Rioja (Northern Spain) during two consecutive years (2003–2005) to study their diversity, occurrence, and life characteristics under different agricultural management representing natural areas and perennial and annual crops from organic and conventional systems. Native EPN species and strains were identified using morphological and molecular characteristics. Virulence, infection cycle length and reproductive potential were assessed using Galleria mellonella larvae. The EPN occurrence was evaluated through abundance, recovery frequency, larval mortality percentage and EPN population density. EPNs were also related to selected soil physical and chemical variables as well as to some soil pollutants such as heavy metals and organochlorine pesticide residues. Only two steinernematids species were identified: Steinernema feltiae was observed throughout all seasons from natural and agricultural areas and Steinernema carpocapsae in summer and autumn of 2004 from perennial crops only. The virulence of native strains was lower than other previously isolated Spanish strain from natural areas or crop field edges. EPN abundance and recovery frequency indicated that habitat type might influence EPNs occurrence stronger than seasonality with the intensity of agricultural management inversely affecting their distribution. Moreover, clay, P₂O₅, Zn, Cu and hexachlorobenzene contents negatively correlating with EPN population density. We consider that agricultural management should be taken into account if EPNs are going to be used as biological control agents.     

Effect of arginine supplementation on the morphology and function of intestinal epithelia and serum concentrations of amino acids in pigs exposed to heat stress

The exposure of pigs to heat stress (HS) appears to damage their intestinal epithelia, affecting the absorption of amino acids (AAs). Arg is involved in the restoration of intestinal epithelial cells but HS reduces Arg intake. The effect of dietary supplementation with Arg on the morphology of intestinal epithelia, AA transporter gene expression, and serum concentration (SC) of free AAs in HS pigs was analyzed. Twenty pigs (25.3 ± 2.4 kg body weight) were randomly assigned to two dietary treatments: Control (0.81% Arg), wheat–soybean meal diet supplemented with l-Lys, l-Thr, dl-Met, and l-Trp, and the experimental diet where 0.16% free l-Arg was supplemented to a similar Control diet (+Arg). All pigs were individually housed and exposed to HS, fed ad libitum with full access to water. The ambient temperature (AT), recorded at 15-min intervals during the 21-d trial, ranged on average from 29.6 to 39.4 °C within the same day. Blood samples were collected on day 18 at 1600 hours (AT peak); serum was separated by centrifugation. At the end of the trial, five pigs per treatment were sacrificed to collect samples of mucosa scratched from each small intestine segment. The expression of AA transporters in intestinal mucosa and the SC of AAs were analyzed. Villi height (VH) was higher (P < 0.01) in the duodenum, jejunum, and ileum but the crypt depth did not differ between the Control and the +Arg pigs. Supplementation of l-Arg increased the mRNA coding for the synthesis of the cationic AA transporter b^0,+ (P < 0.01) and the neutral AA transporter B⁰ (P < 0.05) in the duodenum by approximately 5-fold and 3-fold, respectively, but no effect on mRNA abundance was observed in the jejunum and ileum. The supplementation of l-Arg increased serum Arg, His, Met, Thr, Trp, and urea (P < 0.05) and also tended to increase Val (P < 0.10) but did not affect Ile, Lys, Leu, and Phe. These results indicate that supplementing 0.16% l-Arg to the Control diet may help to improve the function of the small intestine epithelium, by increasing the VH, the abundance of AA transporters, and the SC of most indispensable AAs in pigs exposed to HS conditions. However, the lack of effect of supplemental Arg on both Lys SC and weight gain of pigs suggests that increasing the Lys content in the +Arg diet might be needed to improve the performance of HS pigs.     

Soil nitrous oxide emissions from grassland: Potential inhibitor effect of hippuric acid

In grassland systems, cattle and sheep urine patches are recognized as nitrous oxide (N₂O) emission hot spots due to the high urinary nitrogen (N) concentrations. Hippuric acid (HA) is one of the constituents of ruminant urine that has been reported as a natural inhibitor of soil N₂O emissions. The aim of this study was to examine the potential for elevated ruminant urine HA concentrations to reduce N₂O emissions, in situ, on an acidic heavy clay soil under poorly drained conditions (WFPS > 85%). A randomized complete block design experiment with three replications and four treatments was conducted using the closed‐static‐flux chamber methodology. The four treatments were applied inside the chambers: control with no artificial urine application (C), control artificial urine (U), and enriched artificial urine with two rates of HA (55.8 and 90 mM, U+HA1, U+HA2). Soil inorganic‐N, soil dissolved organic carbon (DOC), soil pH as well as N₂O and methane (CH₄) fluxes were monitored over a 79‐d period. Although N₂O emissions were not affected by the HA enriched urine treatments, U+HA2 positively affected the retention of N as until day 3, when the soil pH dropped to values < 5. Subsequently, as a consequence of rainfall events and soil acidification, it is likely that leaching or sorption onto clay reduced the efficacy of HA, masking any treatment differential effect on N₂O emissions. Moreover, CH₄ fluxes as well as DOC results reflected the soil anaerobic conditions which did not favour nitrification processes. Further research is needed to determine the fate of HA into the soil which might clarify the lack of an in situ effect of this compound.     

Sunflower crop-wild hybrids: Identification and risks

Wild sunflower, Helianthus annuus, is an open-pollinated species that can be a major weed in several crops. If it hybridizes with cultivated sunflower the following generations can generate crop-wild hybrids that could evolve into invasive populations. The objective of this study was to morphologically characterize F1 hybrids between inbred lines and wild sunflower, their F2, BC1 and BC2 progenies with domestic sunflower to assess the risk of these plants evolving into invasive biotypes. Crop-wild hybrids were easily identified as off-type plants. F1 plants were taller, branched with several heads, smaller disks and phyllaries, stigma and pale anthocyanins, and a longer flowering period compared to cultivated sunflower. None of the F2 plants were similar to wild sunflower plants, but morphologically closer to the cultivated materials. These results showed the improbability that crop-wild off-type plants give rise to plants morphologically similar to wild sunflowers and therefore the risk of developing invasive populations by their introduction.     

The world's richest tadpole communities show functional redundancy and low functional diversity: ecological data on Madagascar's stream-dwelling amphibian larvae

Background  

Functional diversity illustrates the range of ecological functions in a community. It allows revealing the appearance of functional redundancy in communities and processes of community assembly. Functional redundancy illustrates the overlap in ecological functions of community members which may be an indicator of community resilience. We evaluated patterns of species richness, functional diversity and functional redundancy on tadpole communities in rainforest streams in Madagascar. This habitat harbours the world's most species-rich stream tadpole communities which are due to their occurrence in primary habitat of particular interest for functional diversity studies.     

Spatial analysis of epidemics of <Emphasis Type="Italic">Grapevine leafroll associated virus-3</Emphasis>

Several vineyards in Rías Baixas and one in the Ribeira Sacra (Spain) were monitored and the spatial pattern of leafroll-diseased grapevine was analysed at several dates. Unidimensional aggregation analysis (ordinary runs), bidimensional analysis, and disease gradients analysis were used as methods of study of spatial aspects of epidemics of GLRaV. At very low insect populations the mealybug Planococcus citri transmitted the GLRaV-3 from infected plants to healthy ones planted between them in an experimental plot at Beluso; an initial focus of leafroll-infected grapevines was detected 2 years after planting at the area where the vectors had been located infesting the old plants. Within 8 years some new foci appeared and coalesced, and the incidence reached >80%. In three commercial plots where no vectors were observed, the spatial analysis of the diseased plants showed three different situations. In Meaño, the study of the evolution of the spatial patterns of diseased plants between 1992 and 2005 suggested slow vectorial field transmission of GLRaV-3. In Goián the analysis for only 2 years suggested random distribution; therefore the viruses were arriving with the planting material, but the runs analysis of some lines suggested incipient spread of GLRaV-3. In Portomarín the incidence of both GLRaV-2 and 3 was low and their distribution was random, without any evidence of field spread. These examples of the study of the spatial analysis of leafroll-infected plants may be helpful to determine whether or not spread of the viruses is occurring, and the best control measures to take.