Efficient analysis of egg yolk proteins and their thermal sensitivity using sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing and nonreducing conditions

The multiple functional properties of egg yolk are mostly influenced by its complex protein composition. The high lipid content of egg yolk as well as the low solubility of delipidated egg yolk lipoproteins make analysis by conventional chromatographic or electrophoretic techniques a difficult task. This work describes a method to profile egg yolk proteins after delipidation with acetone using sodium dodecyl sulfate polyacrylamide gel electrophoresis on precast 8-18% T polyacrylamide gradient gels. Twenty bands were obtained for the whole egg yolk profile with molecular weights ranging between 5 and 221 kDa. The bands were identified based on their molecular weight and by comparison with isolated egg yolk subfractions. The dissociation behavior under reducing and nonreducing conditions provided additionally helpful information for identification and characterization of the yolk proteins. The method presented is very well suited for assaying the thermal sensitivity of whole yolk and its components and thus for the characterization of heat treatment processes.     

Distortion of genetically modified organism quantification in processed foods: influence of particle size compositions and heat-induced DNA degradation

Milling fractions from conventional and transgenic corn were prepared at laboratory scale and used to study the influence of sample composition and heat-induced DNA degradation on the relative quantification of genetically modified organisms (GMO) in food products. Particle size distributions of the obtained fractions (coarse grits, regular grits, meal, and flour) were characterized using a laser diffraction system. The application of two DNA isolation protocols revealed a strong correlation between the degree of comminution of the milling fractions and the DNA yield in the extracts. Mixtures of milling fractions from conventional and transgenic material (1%) were prepared and analyzed via real-time polymerase chain reaction. Accurate quantification of the adjusted GMO content was only possible in mixtures containing conventional and transgenic material in the form of analogous milling fractions, whereas mixtures of fractions exhibiting different particle size distributions delivered significantly over- and underestimated GMO contents depending on their compositions. The process of heat-induced nucleic acid degradation was followed by applying two established quantitative assays showing differences between the lengths of the recombinant and reference target sequences (A, deltal(A) = -25 bp; B, deltal(B) = +16 bp; values related to the amplicon length of the reference gene). Data obtained by the application of method A resulted in underestimated recoveries of GMO contents in the samples of heat-treated products, reflecting the favored degradation of the longer target sequence used for the detection of the transgene. In contrast, data yielded by the application of method B resulted in increasingly overestimated recoveries of GMO contents. The results show how commonly used food technological processes may lead to distortions in the results of quantitative GMO analyses.     

Plant species richness and composition in managed grasslands: The relative importance of field management and environmental factors

Decline of grassland diversity throughout Europe within the last decades is threatening biological diversity and is a major conservation problem. There is an urgent need to determine the underlying factors that control vascular plant species richness and composition in managed grasslands. In this study, 117 grasslands were sampled using standardised methods. Explanatory variables were recorded for each grassland site, reflecting the local field management, site-specific environmental conditions and large-scale spatial trends. Using variation partitioning methods, we determined the pure and shared effects of these three sets of explanatory variables on the plant species richness and composition in grasslands. Most of the explained variation in plant species richness was related to the joint effect of local field management and environmental variables. However, the applied variation partitioning approach revealed that the pure effect of spatial variables contributed relatively little to explaining variation in both the plant species richness and species composition. The largest fractions of explained variation in plant species composition were accounted for by the pure effects of environmental and local field management variables. Moreover, the results revealed that the main mechanisms by which these sets of explanatory variables affect plant species vary according to the type of management regime under study. From our findings we could conclude that particularly a reduction of nitrogen fertilisation on meadows and grazing at a low stocking rate on pastures can help to conserve biodiversity.     

Computerized apparatus for measuring dynamic flavor release from liquid food matrices

A fully computer-controlled apparatus was designed. It combines a glass reactor with a temperature-controlled hood, in which headspace volatiles are captured. Flavored liquids can be introduced into the reactor and exposed to conditions of temperature, air flow, shear rate, and saliva flow as they occur in the mouth. As the reactor is completely filled before measurements are started, creation of headspace just before sampling start prevents untimely flavor release resulting in real time data. In the first 30 s of flavor release the concentrations of the volatiles can be measured up to four times by on-line sampling of the dynamic headspace, followed by off-line trapping of the samples on corresponding Tenax traps and analysis using GC-TDS-FID. Flavor compounds from different chemical classes were dissolved in water to achieve concentrations typically present in food (micrograms to milligrams per liter). Most of the compounds showed constant release rates, and the summed quantities of each volatile of three 10 s time intervals correlated linearly with time. The entire method of measurement including sample preparation, release, sampling, trapping, thermodesorption, and GC analysis showed good sensitivity [nanograms (10 s)(-1)] and reproducibility (mean coefficient of variation = 7.2%).     

Historical Contaminated Sediments and Soils at the River Basin Scale

Background, Aims, and Scope  

Data from the Elbe River and its tributaries indicate, despite extensive improvement in water quality during the last 15 years, that the respective sediment situation of many priority pollutants has not reached an acceptable level. For the coming decades, risks for downstream sites and stakeholders will persist, mainly due to secondary sources originating from historical pollution of soils and sediments in the catchment area. In practice, a catchment-wide assessment of historical contaminated soil and sediment should apply a three-step approach: (i) Identification of substances of concern (s.o.c.) and their classification into ’hazard classes of compounds’; (ii) identification of areas of concern (a.o.c.) and their classification into ‘hazard classes of sites’; (iii) identification of areas of risk (a.o.r.) and their assessment relative to each other with regard to the probability of polluting the sediments in the downstream reaches. The conversion of this concept has to consider the underlying philosophy of the EU Water Framework Directive, particularly with respect to the analysis and monitoring of priority substances in solid matrices. However, major deficiencies are still in the assessment and prognosis of resuspension processes, and potential approaches to fill this gap are described both in theory and from examples of the Elbe River.     

Mineralization of hydroxylated isoproturon metabolites produced by fungi

When exposed to the herbicide isoproturon, some soil fungi in pure culture metabolize the substance to hydroxylated metabolites. Hydroxylated metabolites of isoproturon have also been detected in soil studies. In an agricultural soil not previously exposed to isoproturon we found that the hydroxylated isoproturon metabolite N-(4-(2-hydroxy-1-methylethyl)phenyl)-N′,N′-dimethylurea mineralized faster than both isoproturon and its N-demethylated metabolite N-(4-isopropylphenyl)-N′-methylurea (MDIPU), thus indicating that mineralization of isoproturon is stimulated by fungal hydroxylation in this soil. In soils previously treated with isoproturon, in contrast, isoproturon and both its hydroxylated and demethylated metabolites mineralized at almost the same rate with up to 52% of the ¹⁴C-ring-carbon being degraded to ¹⁴CO₂ within 63 days. Thus hydroxylated metabolites of isoproturon do not seem to be more persistent than isoproturon, and hence may degrade before they can leach from topsoil and contaminate the aquatic environment. While an isoproturon-mineralizing bacterium Sphingomonas sp. SRS2 and a MDIPU-mineralizing mixed bacterial culture were able to deplete the medium of hydroxylated metabolites, little or no mineralization took place. This indicates that other bacteria must be present in the soil that are able to benefit from isoproturon being made available to mineralization by fungal hydroxylation.     

Ökologische Gruppierung von Böden nach ihrem chemischen Bodenzustand

Ecological grouping of soils according to their chemical soil state On the base of the solubility of carbonic acid, calcium carbonate, as well as the oxidic compounds of Al and Fe existing in soils, the following buffer ranges are defined and separated from each other by pH values in the soil solution: Carbonic acid/Carbonate buffer range (pH 6.2 to 8.6), Carbonic acid/Silicate buffer range (pH 5.0 to 6.2), Exchanger buffer range (pH 4.2 to 5.0), Aluminium buffer range (pH 2.8 to 4.2), Iron buffer range (pH 2.4 to 3.8). The buffer capacities are calculated and literature indications regarding buffer rates are compiled. The characteristical chemical soil state is demonstrated for the various buffer ranges with aid of a soil collection. With increasing soil acidification the Ca/Al mole ratio in fine roots decreases from values > 1 to values around 0.3; this is taken as evidence for increasing disposition for Al-toxicity. It is suggested to attach soil horizons to buffer ranges on the base of pH(H₂O) measurements made on field fresh samples. The measurement of pH in salt solution is taken as measure for the amplitude of the chemical soil state in case of H-ion or salt stress.     

Adsorption—desorption and/or precipitation—dissolution processes of zinc in soils

The concentrations of trace and toxic metals in soil solutions are explained by several authors either in terms of adsorption—desorption or precipitation—dissolution reactions in soils. Data have been given for zinc to test the applicability of both concepts. The results show that the concentrations of zinc in equilibrium solutions with soil clay fractions and whole soil samples at pH values below 7 are determined exclusively by adsorption—desorption reactions for various pH's, contents of bound zinc and compositions of soils. At neutral to alkaline pH values precipitation—dissolution reactions of zinc may take place. There is some evidence that formation of zinc silicates may control the zinc concentration in solution provided natural complexing agents are absent, the affinity of the soil for zinc is low and the content of reaching zinc is high (> ～ 100 ppm). Even at pH values above 7, the formation of other zinc compounds is unlikely in most soils because additions of large amounts of zinc are required to ensure saturation of the adsorption sites of different soil components before the zinc concentration in the soil solution can increase sufficiently to bring about the precipitation of definite compounds. Model experiments in CaCO₃-buffered systems showed that the adsorption capacity for specifically adsorbed zinc (in μmole/g) by the following components increased in the order CaCO₃ (0.44), bentonite (44), humic acid (842), amorphous Fe- and Al-oxides (1190, 1310) and δ -MnO₂ (1540) and demonstrated the importance of Mn-, Fe-, and Al-oxides and humic substances for the binding of zinc in soils containing carbonates, and thus indicate the special role of these components in limiting precipitation reactions.