Molecular characterization of Miraflores peach variety and relatives using SSRs

Some traditional peach varieties, originated from the region of Aragón (Spain), were analysed by SSRs (simple sequence repeats). The aim of this research was to characterize 19 clones related to Miraflores variety, with unknown pedigrees, to assess their genetic diversity and to elucidate their possible relationships with 10 traditional peach varieties. Twenty SSR primer pairs with high levels of polymorphism, which have been previously developed for peach, were used in this study. A total of 46 alleles were obtained for all the microsatellites studied, ranging from one to six alleles per locus, with a mean value of 2.3 alleles per locus. Fourteen SSRs were polymorphic in the set of varieties studied and permitted to distinguish 16 different genotypes out of the 30 initially studied, although fourteen ‘Miraflores’ clones showed identical gel profiles. The genetic distance matrix was used to construct neighbor joining cluster and to perform principal coordinate analysis which allowed the arrangement of all the genotypes according to their genetic relationships. The genetic relationships among these traditional peach varieties, and in particular among ‘Miraflores’ clones are discussed. The obtained results confirm that microsatellite markers are very useful for these purpose.     

Performance of a photo‐heterotrophic,hypersaline system for intensive cultivation of white leg shrimp (Litopenaeus vannamei) with minimal water replacement in lined ponds using a stochastic approach

The aim of the study was to evaluate the performance of Litopenaeus vannamei in an intensive photo‐heterotrophic hypersaline system with minimal seawater replacement, and establish relationships between parameters of a stochastic production model and relevant water quality variables. Six experimental 1000 m² lined ponds were stocked at a density of 120 shrimp m^?2 for a 105‐day trial. Salinity increased from 37 to 45 ± 2 g/L, and the water level was maintained with the weekly addition of filtered seawater, equivalent to 1.6% per day. The stochastic model predicted that, at harvest, there is 95% confidence that the system produces between 12.1 and 14.7 t/ha with a mean final individual weight of 13.1 g and a mean survival of 84.2%. Sensitivity analyses showed that dissolved oxygen and individual final weight of shrimp were the main variables influencing yield variance. Nitrogenous compounds were maintained between optimal cultivation levels (NH₃–NH₄⁺ = 0.73 ± 0.43 mg/L, N–NO₂^? = 0.09 ± 0.05 mg/L, N–NO₃^? = 3.22 ± 0.11 mg/L). Heterotrophic bacteria (6.6 ± 3.4 × 10⁵ CFU/ml) and chlorophyll‐α concentration (108.5 ± 80.2 μg/L) showed a similar development pattern, indicating a strong relationship between bacteria and microalgae during cultivation. Vibrio spp. concentrations were low (1.24 ± 1.42 × 10³ CFU/ml). It was shown that the photo‐heterotrophic system could be used in hypersaline conditions, typical of semi‐arid regions, to consistently produce between 12.1 and 14.7 t/ha in 15 weeks.     

Production of monoclonal antibodies to the N-methylcarbamate pesticide propoxur

Monoclonal antibodies (MAbs) to the pesticide propoxur were produced from haptens with carboxylic spacer arms of different lengths introduced at the carbamate group of the analyte structure. MAbs were subsequently characterized in the conjugate-coated format using these immunizing haptens and newly synthesized compounds as homologous and heterologous assay conjugates, respectively. Appropriate combinations of immunoreagents resulted in competitive enzyme immunoassays (ELISA) with I(50) values in the low nanomolar range (6.5-17.9 nM). A modification of the conjugate-coated format consisting of the simultaneous incubation of the MAb and the peroxidase-labeled secondary antibody in the presence of the analyte resulted in an assay with an I(50) value of 4.4 nM. This one-step conjugate-coated ELISA format is as simple and fast as the antibody-coated format but without the need of synthesizing enzyme-hapten conjugates. Major N-methylcarbamate pesticides were not recognized by the MAb. This immunoassay should reasonably allow the rapid, low-cost, and sensitive determination of propoxur in food, soils, and the environment at levels of regulatory and practical importance.     

Climate tolerances of Philaenus spumarius should be considered in risk assessment of disease outbreaks related to Xylella fastidiosa

The bacterium Xylella fastidiosa (Xf) is an invasive insect-borne pathogen, which causes lethal diseases to important crops including olives, citrus, almonds and grapes as well as numerous forest, ornamental, and uncultivated plants. Outbreaks of Xf-related plant diseases are currently occurring in the Mediterranean region, causing substantial losses to various agricultural sectors. Several models have recently been published to identify which regions are at highest risk in Europe; however, such models did not consider the insect vectors, which constitute the key driver of short-range Xf spread. We fitted bioclimatic species distribution models to depict the macroclimatic preferences of the meadow spittlebug Philaenus spumarius L. (1978) (Hemiptera: Aphrophoridae), the major epidemiologically relevant vector currently responsible for Xf spread in the Europe. Many regions of Western Europe and Mediterranean basin are predicted by models as highly climatically suitable for this vector, including all regions where severe Xf have occurred so far. Conversely, the driest and warmest areas of the Mediterranean basin are predicted as little suitable for P. spumarius. Models forecast that agricultural-important parts of the southern Mediterranean area might experience a substantial decrease in climatic suitability for P. spumarius by the period 2040–2060. Areas predicted as highly suitable just for the bacterium but not optimal for this vector are apparently still free of severe Xf outbreaks, suggesting that climate tolerances of P. spumarius might partly explain the current spatial pattern of Xf outbreaks in Europe and should always be considered in further risk assessments.

     

Phospholipid interactions protect the milk allergen alpha-lactalbumin from proteolysis during in vitro digestion

Interactions with food components may alter the resistance of food proteins to digestion, a property thought to play an important role in determining allergenic properties. The kinetics of breakdown of the bovine milk allergen alpha-lactalbumin during in vitro gastrointestinal digestion was found to be altered by interactions with physiologically relevant levels of phosphatidylcholine (PC), a surfactant that is abundant both in milk and is actively secreted by the stomach. Breakdown during gastric digestion was slowed in the presence of PC and accompanied by small alterations in the profile of resulting peptides, with little effect being observed during subsequent duodenal digestion. alpha-Lactalbumin was found to unfold at gastric (acid) pH, giving a CD spectrum similar to that obtained for the partially folded state it is known to adopt at pH values below its isoelectric point. Fluorescence polarization studies performed at low pH indicated that this partially unfolded form of the protein was able to penetrate into the PC vesicles. These interactions are probably responsible for the slowing of gastric digestion by reducing the accessibility of the protein to pepsin. These findings show that interactions with other food components, such as lipids, may alter the rate of breakdown of food proteins in the gastrointestinal tract. It underlines the importance of the food matrix in affecting patterns of food allergen digestion and hence presentation to the immune system and that in vitro digestion systems used for assessing digestibility of allergens must take account of surfactants.     

Detection of anti-Leishmania infantum antibodies in sylvatic lagomorphs from an epidemic area of Madrid using the indirect immunofluorescence antibody test

An outbreak of human leishmaniosis was confirmed in the southwest of the province of Madrid, Spain, between July 2009 and December 2012. Incidence of Leishmania infection in dogs was unchanged in this period, prompting a search for alternative sylvatic infection reservoirs. We evaluated exposure to Leishmania in serum samples from animals in the area with an indirect immunofluorescence test (IFAT). Using promastigotes from six culture passages and a 1/25 threshold titer, we found anti-Leishmania infantum seroreactivity in 9.3% of cats (4 of 43), 45.7% of rabbits (16/35) and 74.1% of hares (63/85). Use of promastigotes from >10 in vitro passages resulted in a notably IFAT lower titer, suggesting antigenic changes during extended culture. Postmortem inspection of seropositive animals showed no clinical signs of infection. The results clearly suggest that asymptomatic hares were the main reservoir in the outbreak, and corroborate IFAT as a sensitive serological surveillance method to detect such cryptic Leishmania infections.     

Correlación Anatomo-radiológica de la Cabeza en la Cabra de Raza Canaria (Capra hircus L.)

A radiologic and tomographic study was made of the cranioencephalic topography of canary goats of medium weight and size. In a first step, the exocranialis references corresponding to an axis designed at the level of the meatus acusticus externus and parallel to the basicranial axis were anatomically and radiologically defined. Conventional stereotaxic techniques were then used to reach the standard position of the head in order to obtain macroscopical cross sections at different levels perpendicular to the base of the cranium. In a second step, a tomographic study was made to confirm the design of the axis.     

Effect of single Quercus ilex trees upon spatial and seasonal changes in soil water content in dehesas of central western Spain

The spatial and temporal evolution of soil water content (θ) in Quercus ilex dehesas has been investigated to determine how trees modify the soil water dynamics and the nature of tree-grass interactions in terms of soil water use in these ecosystems. Soil physical parameters and θ were measured at different distances from the tree trunk (2–30 m) in the upper 300 cm of soil. θ was measured monthly by TDR during 2002–2005. Tree water potential was determined during the summers of 2004 and 2005. At deeper soil layers, mean θ values were higher beyond than beneath tree canopy during dry periods. θ depletion beyond tree canopy continued even in summer, when herbaceous plants dried up, suggesting that trees uptake water from the whole inter-tree space. Results have shown a high dependence of trees on deep water reserves throughout late spring and summer, which helps to avoid competition for water with herbaceous vegetation.