Evaluation of the antioxidant activity of capsiate analogues in polar, nonpolar, and micellar media

Synthesis of 10 capsiate analogues was conducted by lipase-mediated (Novozyme 435) esterification of vanillyl alcohol with different fatty acids. The antioxidant activity of the synthesized capsiates was evaluated using three in vitro assays: DPPH radical scavenging assay (polar medium), Rancimat assay (nonpolar medium), and autoxidation of linoleic acid (micellar medium). The objective of this study is to find the influence of structural characteristics of the alkyl chain of capsiate analogues on their antioxidant activity. In these assays, BHT and α-tocopherol were used as reference compounds. Both DPPH and Rancimat assays did not show any specific trend of antioxidant activity with the increase in lipophilicity and also with the type of fatty acids grafted to the phenolic moiety. In the Tween 20 micellar system for the inhibition of autoxidation of linoleic acid, vanillyl ester attached to a C18 alkyl chain (vanillyl stearate, oleate, and ricinoleate) exhibited maximum inhibition of autoxidation of linoleic acid.     

Biofortification of soybean meal: immunological properties of the 27 kDa γ-zein

Legumes, including soybeans ( Glycine max ), are deficient in sulfur-containing amino acids, which are required for the optimal growth of monogastric animals. This deficiency can be overcome by expressing heterologous proteins rich in sulfur-containing amino acids in soybean seeds. A maize 27 kDa γ-zein, a cysteine-rich protein, has been successfully expressed in several crops including soybean, barley, and alfalfa with the intent to biofortify these crops for animal feed. Previous work has shown that the maize 27 kDa zein can withstand digestion by pepsin and elicit an immunogenic response in young pigs. By use of sera from patients who tested positive by ImmunoCAP assay for elevated IgE to maize proteins, specific IgE binding to the 27 kDa γ-zein is demonstrated. Bioinformatic analysis using the full-length and 80 amino acid sliding window FASTA searches identified significant sequence homology of the 27 kDa γ-zein with several known allergens. Immunoblot analysis using human serum that cross-reacts with maize seed proteins also revealed specific IgE-binding to the 27 kDa γ-zein in soybean seed protein extracts containing the 27 kDa zein. This study demonstrates for the first time the allergenicity potential of the 27 kDa γ-zein and the potential that this protein has to limit livestock performance when used in soybeans that serve as a biofortified feed supplement.     

Comparison of the results of soil profiles’ diagnostics performed in three classification systems

Three soil classification systems—the World Reference Base for Soil Resources (WRB), Soil Taxonomy, and the recent Russian system—were used for the identification of 17 soil profiles in southwestern Poland; all the systems put emphasis on the soil properties as diagnostic criteria. Different soils developed on glaciofluvial plains, loessic uplands, and in the Sudetes Mountains were classified. The best correlation between the classification decisions in the different systems was obtained for the most widespread soils owing to the similarity of the diagnostic criteria, which were essentially close although not coinciding. The most prominent divergence between the systems in both the names and the taxonomic categories of the soils was found for the polygenetic soils and for the soils developing from the lithologically discontinuous parent materials. It was also found that the diagnostic elements differ in terms of their taxonomic importance among the classification systems.     

土壤持水特性参数与土壤质地尺度关系的联合多重分形分析

Soil water-retention characteristics at measurement scales are generally different from those at application scales, and there is scale disparity between them and soil physical properties. The relationships between two water-retention parameters, the scaling parameter related to the inverse of the air-entry pressure (αvG, cm-1) and the curve shape factor related to soil pore-size distribution (n) of the van Genuchten water-retention equation, and soil texture (sand, silt, and clay contents) were examined at multiple scales. One hundred twenty-eight undisturbed soil samples were collected from a 640-m transect located in Fuxin, China. Soil water-retention curves were measured and the van Genuchten parameters were obtained by curve fitting. The relationships between the two parameters and soil texture at the observed scale and at multiple scales were evaluated using Pearson correlation and joint multifractal analyses, respectively. The results of Pearson correlation analysis showed that the parameter αvG was significantly correlated with sand, silt, and clay contents at the observed scale. Joint multifractal analyses, however, indicated that the parameter αvG was not correlated with silt and sand contents at multiple scales. The parameter n was positively correlated with clay content at multiple scales. Sand content was significantly correlated with the parameter n at the observed scale but not at multiple scales. Clay contents were strongly correlated to both water-retention parameters because clay content was relatively low in the soil studied, indicating that water retention was dominated by clay content in the field of this study at all scales. These suggested that multiple-scale analyses were necessary to fully grasp the spatial variability of soil water-retention characteristics.     

Species composition of arbuscular mycorrhizal fungi in two mountain meadows with differing management types and levels of plant biodiversity

Species composition of arbuscular mycorrhizal fungi (AMF) was analysed in two differently managed mountain grasslands in Thuringia (Germany). Arbuscular mycorrhizal fungi were studied in the roots of 18 dominant plant species from a total of 56 (32%). Additionally, spores of AMF were isolated from soil samples. Arbuscular mycorrhizal fungi species composition was analysed based on 96 sequences of the internal transcribed spacer of the nuclear ribosomal DNA, 72 originated from mycorrhizal roots, and 24 originated from AMF spores. Phylogenetic analyses revealed a total of 19 AMF species representing all genera of the Glomeromycota except Scutellospora and Pacispora. Despite a different farming intensity, resulting in remarkable differences concerning their plant species diversity (27 against 43 plant species), the diversity of AMF was found to be similar with 11 species on the intensively farmed meadow and ten species on the extensively farmed one. Nevertheless, species composition between both sites was clearly different. It thus seems likely that the AMF species composition, but not necessarily the species number, is related to above ground plant biodiversity in the system under study.     

Microbial community structure in forest soils treated with a fire retardant

The influence of a fire retardant (Firesorb, an acrylic–acrylamide copolymer) on the microbial community structure determined by phospholipid fatty acid (PLFA) analysis was examined under laboratory conditions using two different textured soils under pine forest. Firesorb was added to unheated and heated soil samples (350°C for 10 min followed by reinoculation, to mimic a forest fire) at three levels of application (none, usual and three times the usual levels), and measurements were made after 12 weeks of incubation. The relative importance of the three factors considered on the PLFA profiles was as follows: soil heating ≫ soil texture ≈ Firesorb treatment. In the unheated soils, Firesorb had a larger effect than soil texture, while the opposite was found in the heated soils. Soil heating reduced the total PLFAs, while Firesorb tended to increase them in both the unheated and heated soils. Soil heating decreased the PLFAs indicative of gram-positive (G⁺) bacteria and tended to increase the fatty acids associated with gram-negative (G⁻) bacteria and, to a lesser extent, the PLFA 18:2ω6, considered to be predominantly of fungal origin. Firesorb treatment decreased the G⁻/G⁺ bacteria ratio in the heated soils but tended to increase it in the unheated soils, the effect being dose dependent.     

Uptake of veterinary medicines from soils into plants

Medicines play an important role in the treatment and prevention of disease. Whereas the side effects on human and animal health resulting directly from treatment have been widely documented, only recently have the occurrence and fate of medicines in the environment and the potential consequences for human health been recognized as an issue warranting consideration. Medicines have been shown to be released to soils and to persist in the environment. This study was performed to investigate the potential for a range of veterinary medicines to be taken up from soil by plants used for human consumption and to assess the potential significance of this exposure route in terms of human health. Soil analyses indicated that, for selected substances, measurable residues of these are likely to occur in soils for at least 5 months following application of manure containing these compounds. Experimental studies on the uptake of veterinary medicines into carrot roots (tubers) and lettuce leaves showed that only florfenicol, levamisole, and trimethoprim were taken up by lettuces, whereas diazinon, enrofloxacin, florfenicol, and trimethoprim were detected in carrot roots. Measured concentrations in plant material were used to model potential adult human exposure to these compounds. Although exposure concentrations were appreciable in a few instances, accounting for approximately 10% of the acceptable daily intake values (ADI), all were lower than the ADI values, indicating that, at least for compounds with properties similar to those considered here, there is little evidence of an appreciable risk. This exposure route may, however, be important when veterinary medicines have a very low ADI, at which they elicit subtle effects over prolonged periods, or when exposure is occurring via a number of routes at once. Although degradation products (produced in the soil or the plant) were not measured, it is possible for some substances that these could increase the risks to consumers.     

Occurrence of Polycyclic Musks in Sewage Sludge and their Behaviour in Soils and Plants. Part 1: Behaviour of Polycyclic Musks in Sewage Sludge of Different Treatment Plants in Summer and Winter (5 pp)

Background, Aim and Scope   Part 1: Behaviour of Polycyclic Musks in Sewage Sludge of Different Treatment Plants in Summer and Winter Part 2: Investigation of Polycyclic Musks in Soils and Plants -  Preamble. In Part 1 of the study, screening tests were performed to investigate the occurrence of PCMs in sewage sludges. For a preliminary risk assessment, further information is needed about their behaviour in the terrestrial environment. Hence, Part 2 examined the adsorption of PCMs to soil, their dissipation and leaching in soil and their uptake by plants. Background, Aim and Scope   Polycyclic Musks (PCMs) enter the environment via the waste water system. Because of their persistence, they can accumulate in different matrices like sewage sludge or biota. By the use of sewage sludge as a fertilizer, PCMs are transferred to agricultural soils. Therefore, in Part 1 of the study, screening tests were performed to investigate the occurrence of PCMs in sewage sludge. For a preliminary risk assessment, further information is needed about their behaviour in the terrestrial environment. Hence, Part 2 of the study examined the adsorption of PCMs to soil, their dissipation and leaching in soil, and their uptake by plants. Materials and Methods: In the screening study, samples of activated sewage sludge were taken both in summer and in winter at 21 treatment plants. In order to get an overview of the contamination situation, sampling covered different types of treatment plants (in rural, urban, industrial areas). Analytical methods for the determination of HHCB, AHTN, ADBI, ATTN, AHDI and ATII in the sludge samples were developed and applied. Results: The analytical screening of PCMs showed their presence in activated and dried sewage sludge samples. HHCB and AHTN represented about 95% of the PCMs investigated. Their concentrations in the activated sludge samples varied between 2.9 and 10.4 mg/kg dry mass (dm) and 1.1 to 4.2 mg/kg dm, respectively. Although different types of sewage treatment plants were investigated, similar PCM levels were found, showing the widespread input of these compounds into domestic waste water. Discussion: PCM concentrations in activated sludge varied widely. The variation drops substantially when concentrations are related to the varying dry mass. In dehydrated sludge, PCM concentrations were up to 24 mg/kg dm for HHCB and up to 6.9 mg/kg dm for AHTN. These high values are comparable to those obtained in other investigations analysing PCMs. If the degradation of organic mass during anaerobic decomposition is included in the evaluation, the figures obtained are comparable to those for activated sludge. Elimination in sewage sludge was higher in summer than in winter. Therefore, the contamination of the sludges in winter reached higher levels compared to the summer. Conclusions: The results show that PCMs are widespread contaminants in sewage sludge. Recommendations and Perspectives: PCM should be considered in a risk assessment as potential contaminants of sewage sludge destined for agricultural use. Due to the high PCM levels in sewage sludge, further investigations into the degradation and elimination behaviour in sewage sludge have to be carried out, including that involving PCM metabolites such as lactone derivatives.     

Sewage Treatment in a Rotating Biological Contactor (RBC) System

The treatment of domestic wastewater at a temperature of 12–24°C was investigated in an RBC system. The RBC consists of a two stage system connected in series. The system was operated at different organic loading rates (OLR's) and hydraulic retntion times (HRT's) in order to optimize the RBC performance. The overall removal efficiencies for chemical oxygen demand (COD_total, COD_suspended and COD_colloidal) significantly decreased when decreasing the total HRT from 10 to 2.5 h and increasing the OLR from 11 to 47 g COD/m².d. However, the effluent quality of COD_soluble remained unaffected. Most of the COD was removed in the 1^st stage and nitrification took place in the 2^nd stage of the two stage system.The overall nitrification efficiency was 49% at total OLR of 11 gCOD/m².d. At total HRT's of 10, 5 and 2.5 h, the Escherchia coli (E. coli) concentration was reduced by a value of 1.6, 1.5 and 0.8 log₁₀ respectively. The sludge volume index (SVI) decreased as the OLR increased. However, the SVI of the excess sludge produced in the RBC under different OLR's was always <74 ml/gTS, which indicates vadjust a good settleability. The performance of the single versus two stage RBC operated at the same total OLR of 22 g COD/m².d and the same HRT of 5.0 h was examined. The results obtained showed that the COD concentration and the E. coli content in the final effluent of a two stage were lower than in the effluent of the single stage RBC. Moreover, the nitrification efficiency in the two stage system was higher comapred to one stage system.     

Identification of glycinin and beta-conglycinin subunits that contribute to the increased protein content of high-protein soybean lines

Seed protein concentration of commercial soybean cultivars calculated on a dry weight basis ranges from approximately 37 to 42% depending on genotype and location. A concerted research effort is ongoing to further increase protein concentration. Several soybean plant introductions (PI) are known to contain greater than 50% protein. These PIs are exploited by breeders to incorporate the high-protein trait into commercial North American cultivars. Currently, limited information is available on the biochemical and genetic mechanisms that regulate high-proteins. In this study, we have carried out proteomic and molecular analysis of seed proteins of LG00-13260 and its parental high-protein lines PI 427138 and BARC-6. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis revealed that the high-protein lines accumulated increased amounts of beta-conglycinin and glycinins, when compared with Williams 82. High-resolution two-dimensional electrophoresis utilizing pH 4-7 and pH 6-11 ampholytes enabled improved resolution of soybean seed proteins. A total of 38 protein spots, representing the different subunits of beta-conglycinin and glycinin, were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. High-protein was correlated with an increase in the accumulation of most of the subunits representing beta-conglycinin and glycinin. Comparisons of the amino acid profiles of high-protein soybean lines revealed that the concentration of sulfur amino acids, a reflection of protein quality, was not influenced by the protein concentration. Southern blot analysis showed the presence of genotypic variation at the DNA level between PI 427138 and BARC-6 for the genes encoding group1 glycinin, beta-conglycinin, Bowman-Birk inhibitor (BBI), and the Kunitz trypsin inhibitor (KTI). LG00-13260 inherited the allelic variants of the parental line PI 427138 for glycinin, beta-conglycinin, and KTI, while BBI was inherited from the parental line BARC-6. The results of our study indicate that high-seed protein concentration is attributed to greater accumulation of specific components of beta-conglycinin and glycinin subunits presumably mediated by preferential expression of these genes during seed development.