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.     

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.     

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.     

Fast and efficient discrimination of the Isotoma viridis group (Insecta: Collembola) by PCR-RFLP

Identification of collembolan species is generally based on specific morphological characters, such as chaetotaxy and pigmentation pattern. However, some specimens do not match to described characters because these refer to adult specimens, often of one specific sex, or the characters are highly variable in adults (e.g. pigmentation, setae or furcal teeth). Isozymes have frequently assisted species discrimination, and also these may vary with developmental stage or environmental conditions. For identification of single species of the Isotoma viridis group, we present both direct sequencing of the cytochrome oxidase subunit II (COII) gene and a simple DNA-based molecular method.

Five PCR primers amplifying the COII region (717 bp) of the mitochondrial DNA were used. The sequences clearly separated the species I. viridis, I. riparia and I. anglicana, irrespective of colour varieties within the first species. DNA amplification products of different species can also be distinguished by digestion with restriction endonucleases, followed by gel electrophoresis for separation of fragments. This restriction fragment length polymorphism (RFLP), obtained after digestion with the endonucleases TaqI, VspI, MvaI and Bsp143I, revealed specific fragments that separated the three species from each other. Since restriction enzymes are sensitive to single base mutations, we suggest to use a combination of enzymes with at least two species-specific restriction sites when using the RFLP technique. For the I. viridis complex, VspI and Bsp143I appear to be an appropriate combination.     

Effects of input management and crop diversity on non-renewable energy use efficiency of cropping systems in the Canadian Prairie

Although producers’ prime objective may be to increase net returns, many are also interested in conserving and enhancing the quality the soil, water and air resources through adopting more environmentally friendly production practices. This study compared non-renewable energy inputs, energy output, and energy use efficiency of nine dryland cropping systems comprised of a factorial combination of three methods of input management [high (HIGH), i.e., conventional tillage plus full recommended rates of fertilizer and pesticides; reduced (RED), i.e., conservation tillage plus reduced rates of fertilizer and pesticides; and organic (ORG), i.e., conventional tillage plus N-fixing legumes and non-chemical means of weed and pest control]; and three crop rotation systems with varying levels of cropping diversity [a fallow-based rotation with low crop diversity (LOW); a diversified rotation using annual cereal, oilseed and pulse grains (DAG); and a diversified rotation using annual grains and perennial forages (DAP)]. The study was conducted over the 1996–2007 period on a Dark Brown Chernozemic soil (Typic Boroll) in the Canadian Prairies. As expected, total direct plus indirect energy input was the highest for the HIGH and RED input treatments (3773 MJ ha^?1 year^?1), and 50% less for ORG management. Most of the energy savings came from the non-use of inorganic fertilizers and pesticides in the ORG management treatments. Further, total energy use was the highest for the DAG treatments (3572 MJ ha^?1 year^?1), and similar but about 18% lower for the DAP and LOW crop diversity treatments compared to DAG. Thus, overall, the HIGH/DAG and RED/DAG systems had the highest energy requirements (4409 MJ ha^?1 year^?1) and ORG/DAP had the lowest (1806 MJ ha^?1 year^?1). Energy output (calorimetric energy content) was typically the highest for the HIGH input treatments (26,541 MJ ha^?1 year^?1), was about 4% less with RED, and 37% less with ORG management. The latter reflected the lower crop yields obtained with organic management. Similarly, energy output was the highest for the DAP treatments (25,008 MJ ha^?1 year^?1), about 5% less for DAG, and 20% less for the LOW crop diversity treatments. The higher energy output with the DAP treatments largely reflected that the entire harvested biomass of the forage crops was included in energy output, while for grain crops only the seed was included. The straw and crop residues from annual crops were returned to the land to protect the soil from erosion and to maintain soil organic matter as this is the recommended practice in this semi-arid region. In contrast to energy output and to net energy produced, energy use efficiency (measured as yield of grain plus forage produced per unit of energy input or as energy output/energy input ratio) was the highest for the ORG input treatments (497 kg of harvested production GJ^?1 of energy input, and an energy output/energy input ratio of 8.8). We obtained lower, but generally similar energy use efficiency for the HIGH and RED input treatments (392 kg GJ^?1 and ratio of 7.1). Thus, overall, ORG/DAP was the most energy efficient cropping system, while RED/LOW and RED/DAG generally ranked the lowest in energy use efficiency. Our findings support the current movement of producers toward ORG management as a means of reducing the reliance on non-renewable energy inputs and improving overall energy use efficiency of their cropping systems. Our results also suggest that moving away from traditional monoculture cereal rotations that employ frequent summer fallowing, toward extended and diversified crop rotations that use reduced tillage methods, although resulting in an increase in energy output, will not significantly reduce the overall reliance on non-renewable energy inputs, nor enhance energy use efficiency, unless perennial legume forages and/or legume grain crops are included in the cropping mix.