Quantitative sandwich ELISA for the determination of tropomyosin from crustaceans in foods

The ubiquitous muscle protein tropomyosin has been identified as the major shrimp allergen and is suggested to be a cross-reacting allergen. Previously, only a few methods for the detection of tropomyosin in food have been published. A quantitative sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of tropomyosin from crustaceans in foods has been developed and validated. A polyclonal rabbit antitropomyosin capture antibody and the biotinylated conjugate of the same antibody for detection were the basis for the ELISA, which was specific for crustaceans. The ELISA was able to quantitate tropomyosin in various food matrixes, had a detection limit of 1 microg/g, and cross-reacted to some extent with cockroach. Recoveries ranged from 63 to 120%, and the intra and interassay coefficients of variation were <6 and <14%, respectively.     

Effects of photoperiod on sex steroids and gonad maturation in Arctic charr

Four groups of Arctic charr (age 3+), previously exposed to natural photoperiod, were subjected to different photoperiod regimes from February 20: Simulated natural photoperiod (SNP; Tromsø 70°N), continuous light (LD 24:0) followed by short day (LD 6:18) from either May 1 (L_Feb–May) or June 25 (L_Feb–June), or continuous light (LD 24:0) throughout the experiment (L_Continuous). In females, peak levels of estradiol-17β and testosterone were 6 and 10, 6 and 7, and 3 and 5 weeks advanced, in L_Feb–May, L_Feb–June and L_Continuous, respectively, whereas in males plasma levels of 11-ketotestosterone peaked 8 weeks earlier in L_Feb–May and L_Feb–June than in SNP and L_Continuous. Median ovulation time in L_Feb–May and L_Feb–June was advanced by 10 weeks compared to SNP and L_Continuous, and median spermiation time (first observation of running milt in each individual) in L_Feb–May, L_Feb–June and L_Continuous was advanced by 10, 7 and 5 weeks, compared to SNP. A switch from long to short day early in the reproductive cycle (L_Feb–May and L_Feb–June) apparently resulted in stronger synchronization of both ovulation and spermiation. Ovulation in L_Feb–May and L_Feb–June occurred over 3 weeks (L_Feb–May) and 8 weeks (L_Feb–June), as compared to 10 weeks in the SNP group, whereas all males (except one individual) began spermiating over a period of 3 weeks (L_Feb–June), or less (L_Feb–May), as compared to 7 weeks in the SNP group. Lack of a short day stimulus (L_Continuous), on the other hand, resulted in larger variation in the timing of final maturation, with ovulation and spermiation being spread over 15 weeks. Low egg survival in L_Feb–May (17% versus 60% and 97% for L_Continuous and SNP) indicate that a long day signal of 10 weeks early in the reproductive cycle may be too short to ensure good egg and sperm quality. Photoperiod manipulation did not affect the proportion of maturing fish.     

Negligible contribution from roots to soil-borne phospholipid fatty acid fungal biomarkers 18:2ω6,9 and 18:1ω9

The phospholipid fatty acid biomarkers 18:1ω9, 18:2ω6,9 and 18:3ω3,6,9 are commonly used as fungal biomarkers in soils. They have, however, also been found to occur in plant tissues, such as roots. Thus, the use of these PLFAs as fungal biomarkers in sieved soil, which may still contain small remains of roots, has been questioned. We used data from a recent beech tree girdling experiment to calculate the contribution of roots to these biomarkers and were able to demonstrate that not more than 0.61% of 18:1ω9 and 18:2ω6,9 in sieved soil samples originated from roots (but 4% of 18:3ω3,6,9). Additionally, the abundance of the biomarker 18:2ω6,9 in the soil was found to be highly correlated to ectomycorrhizal root colonization, which further corroborates its fungal origin. PLFA biomarkers were substantially reduced in vital roots from girdled trees compared to roots of control trees (by up to 76%), indicating that the major part of PLFAs measured in roots may actually originate from ectomycorrhizal fungi growing inside the roots. We calculated, that even a near to 50% reduction in fine root biomass - as observed in the girdling treatment - accounted for only 0.8% of the measured decrease of 18:2ω6,9. Our results demonstrate that both 18:1ω9 and 18:2ω6,9 are suitable biomarkers for detecting fungal dynamics in soils and that especially 18:2ω6,9 is a reliable biomarker to study mycorrhizal dynamics in beech forests.     

Incidental phosphorus losses – are they significant and can they be predicted?

Fresh applications of phosphorus (P) may cause ‘incidental’ losses of dissolved and particulate P forms in land runoff when rainfall interacts directly with fertilizers and manures which are spread, or excreted, onto the soil surface. Research indicates that when incidental P losses (IPL) occur, they often make the dominant (50–98 %) contribution to measured P loads in surface and sub‐surface runoff from field plots, with increased risk of eutrophication where P‐enriched runoff is routed directly to the watercourse. Rates of P loss are temporally and spatially very variable (< 1 to 25 % of total P applied) depending on the amount of P applied; the P release properties of the materials applied (% P extractable in water), the timing of storm events after application and the amounts of runoff generated. Large P applications left on the surface of wet, frozen, compacted, and intensively underdrained soils in high rainfall areas are particularly vulnerable to IPL. Concentrations of P in runoff are often greatest during the first storm event following P application, but can remain high for several weeks, or even months after application. Catchment scale impacts are more difficult to quantify due to the effects of dilution with runoff from fields without IPL, and the contributions of P from other sources. Approaches to predicting IPL are discussed and need to be improved in order to assess the contribution of IPL to catchment P loads in river basin management planning. Strategies to control IPL should focus on adoption of more sensitive management practices in fields with a high risk of rapid runoff or preferential flow, and are likely to very effective.     

The conceptual basis for a decision support framework to assess the risk of phosphorus loss at the field scale across Europe

We report a conceptual framework for a simple screening tool to evaluate the vulnerability of different fields in the landscape to phosphorus (P) loss within the European agricultural context. Our work builds on the principles established for the P Index system developed in the United States. We examine the differing needs and criteria for a range of EU member states and EFTA countries and highlight essential and optional criteria for inclusion in the decision support framework depending on their ecoregional characteristics. A framework such as this, meets in principle, the needs of the COST832 Action ‘Quantifying the Agricultural Contribution to Eutrophication’ as it deals with diffuse nutrient sources from agriculture, and aids more targeted use of practical strategies to mitigate nutrient losses to minimise the potential for the eutrophication of receiving waters.     

Degradation and binding of atrazine in surface and subsurface soils

Understanding the dissipation rates of chemicals in unsaturated and saturated zones of subsurface soils will help determine if reductions of concentrations to acceptable levels will occur. Chemical properties and microbial biomass and activity were determined for the surface (0-15 cm), lower root (50-105 cm), and vadose (175-220 cm) zones in a Huntington silty clay loam (Fluventic Hapludoll) collected from an agricultural field near Piketon, OH. The rates of sorption, mineralization, and transformation (formation of bound residues and metabolites) of atrazine were determined. Microbial activity was estimated from the mineralization of (14)C-benzoate. We observed decreased levels of nutrients (total organic carbon, N, and P) and microbial biomass with depth, while activity as measured with benzoate metabolism was higher in the vadose zone than in either the surface or the root zones. Sorption coefficients (K(f)) declined from 8.17 in the surface to 3.31 in the vadose zone. Sorption was positively correlated with organic C content. Rates of atrazine mineralization and bound residues formation were, respectively, 12-2.3-fold lower in the vadose than in the surface soil. Estimated half-lives of atrazine ranged from 77 to 101 days in the surface soil, but increased to over 900 days in the subsurface soils. The decreased dissipation of atrazine with increasing depth in the profile is the result of decreased microbial activity toward atrazine, measured either as total biomass or as populations of atrazine-degrading microorganisms. The combination of reduced dissipation and low sorption indicates that there is potential for atrazine movement in the subsurface soils.     

Effect of source germplasm and season on the in vivo haploid induction rate in tropical maize

For in vivo production of doubled haploid (DH) lines in maize, the rate of haploid induction is of crucial importance. Maternal haploid induction depends primarily on the inducer used as a pollinator. However, the source germplasm used as a maternal parent and the environmental conditions for induction may also influence haploid induction and these aspects have not been examined in tropical maize so far. The objectives of our study were to (i) monitor the variation for haploid induction rate (HIR) among diverse source germplasm in tropical maize, (ii) determine the relative importance of general (GCA) and specific (SCA) combining abilities for HIR, and (iii) investigate the influence of summer and winter seasons and genotype × season interactions on this trait. Ten inbreds were mated in a half diallel design. The resulting 45 F₁ single crosses were pollinated with the haploid inducer hybrid RWS × UH400 during the summer 2008 and winter 2009 seasons in a lowland tropical environment in Mexico. HIR of the single crosses averaged over seasons ranged from 2.90 to 9.66% with an overall mean of 6.74%. Mean HIR was significantly (P < 0.01) higher during the winter (7.37%) than summer season (6.11%). Significant (P < 0.01) variation was observed due to GCA effects of parental inbreds of single crosses but not for SCA, GCA × season and SCA × season interactions. Our study underpins that a higher HIR in tropical maize can be obtained by selecting appropriate source germplasm and undertaking pollination under favorable environmental conditions.     

CX3CR1-mediated dendritic cell access to the intestinal lumen and bacterial clearance

Dendritic cells (DCs) and macrophages are critical to innate and adaptive immunity to the intestinal bacterial microbiota. Here, we identify a myeloid-derived mucosal DC in mice, which populates the entire lamina propria of the small intestine. Lamina propria DCs were found to depend on the chemokine receptor CX3CR1 to form transepithelial dendrites, which enable the cells to directly sample luminal antigens. CX3CR1 was also found to control the clearance of entero-invasive pathogens by DCs. Thus, CX3CR1-dependent processes, which control host interactions of specialized DCs with commensal and pathogenic bacteria, may regulate immunological tolerance and inflammation.     

Reward-predictive cues enhance excitatory synaptic strength onto midbrain dopamine neurons

Using sensory information for the prediction of future events is essential for survival. Midbrain dopamine neurons are activated by environmental cues that predict rewards, but the cellular mechanisms that underlie this phenomenon remain elusive. We used in vivo voltammetry and in vitro patch-clamp electrophysiology to show that both dopamine release to reward predictive cues and enhanced synaptic strength onto dopamine neurons develop over the course of cue-reward learning. Increased synaptic strength was not observed after stable behavioral responding. Thus, enhanced synaptic strength onto dopamine neurons may act to facilitate the transformation of neutral environmental stimuli to salient reward-predictive cues.