Long-Term Soil Reaction Changes in a Temperate Beech Forest Subject to Past Alkaline Pollution

Changes in deposition exert effects on forests. Some evidence for a slow recovery of strongly alkalised forest soils after a reduction in alkaline pollution exists, but the recovery rate is little known. The objective of this study was to estimate temporal soil reaction changes. For this purpose, measurements of soil reaction under a beech forest near magnesite works in Central Slovakia were taken in 1991 and 2006, following the alkaline pollution reduction in 1992. As a result, the mean active soil reaction decreased from pH 7.8 to 6.6. Beyond the range of beech stemflow, which amplifies water and acid pollution inputs into soils locally, soil reaction dropped from pH 7.8 to 6.8. The effect of stemflow on local exchangeable reaction minima was detected even before the pollution reduction. The logarithmic function fitted on the pH data indicates considerable differences between neutralisation rates within and outside the stemflow zone.     

Characterization of the inflammatory infiltrate and cytokine expression in the skin of horses with recurrent urticaria

Background – Recurrent urticaria (RU) is a common skin disease of horses, but little is known about its pathogenesis. Hypothesis/Objective – The aim of this study was to characterize the inflammatory cell infiltrate and cytokine expression pattern in the skin of horses with RU. Animals – Biopsies of lesional and nonlesional skin of horses with RU (n = 8) and of skin from healthy control horses (n = 8) were evaluated. Methods – The inflammatory cell infiltrate was analysed by routine histology. Immunohistochemistry was used to identify T cells (CD3), B cells (CD79), macrophages (MAC387) and mast cells (tryptase). Expression of T‐helper 2 cytokines (interleukins IL‐4, IL‐5 and IL‐13), a T‐helper 1 cytokine (interferon‐γ), IL‐4 receptor α and thymic stromal lymphopoietin was assessed by quantitative RT‐PCR. Results – In subepidermal lesional skin of RU‐affected horses, increased numbers of eosinophils (P ≤ 0.01), CD79‐positive (P ≤ 0.01), MAC387‐positive (P ≤ 0.01) and tryptase‐positive cells (P ≤ 0.05) were found compared with healthy horses. Subepidermal lesional skin of RU‐affected horses contained more eosinophils (P ≤ 0.05) and tryptase‐positive cells (P ≤ 0.05) compared with nonlesional skin. There was no significant difference in infiltrating cells between nonlesional skin and skin of healthy horses. Expression of IL‐4 (P ≤ 0.01), IL‐13 (P ≤ 0.05), thymic stromal lymphopoietin (P ≤ 0.05) and IL‐4 receptor α (P ≤ 0.05) was increased in lesional skin of RU‐affected horses compared with control horses. Expression of IL‐4 was higher (P ≤ 0.05) in lesional compared with nonlesional RU skin. Conclusions and clinical importance – Analysis of cytokine expression and inflammatory infiltrate suggests that T‐helper 2 cytokines, eosinophils, mast cells and presumptive macrophages play a role in the pathogenesis of equine RU.     

ParaCalc®--a novel tool to evaluate the economic importance of worm infections on the dairy farm

Subclinical infections with gastrointestinal nematodes and liver fluke are important causes of production losses in grazing cattle. Although there is an extensive compilation of literature describing the effect of these infections on animal performance, only a few attempts have been made to convert these production losses to an economic cost. Here, we propose a novel tool (ParaCalc(?)), available as a web-application, to provide herd-specific estimates of the costs of these infections on dairy farms. ParaCalc(?) is a deterministic spread-sheet model where results from diagnostic methods to monitor the helminth infection status on a herd and anthelmintic usage are used as input parameters. Default values are provided to describe the effects of the infections on production and the cost of these production losses, but the latter can be adapted to improve the herd-specificity of the cost estimate. After development, ParaCalc(?) was applied on input parameters that were available for 93 Belgian dairy herds. In addition, the tool was provided to 6 veterinarians and their user experiences were evaluated. The estimated median [25th-75th percentile] cost per year per cow was € 46 [29-58] and € 6 [0-19] for gastrointestinal nematode and liver fluke infection, respectively. For both infections, the major components in the total costs were those associated with milk production losses in the adult cows. The veterinarians evaluated ParaCalc(?) as a useful tool to raise the farmers' awareness on the costs of worm infections, providing added value for their services. However, the score given for user-friendliness was diverse among users. Although the model behind ParaCalc(?) is a strong simplification of the real herd processes inducing economic losses, the tool may be used in the future to support economic decisions on helminth control.     

Generation of equine TSLP-specific antibodies and their use for detection of TSLP produced by equine keratinocytes and leukocytes

Allergic horses react to innocuous environmental substances by activation of Th2 cells and production of allergen-specific IgE antibodies. The mechanisms leading to Th2 differentiation are not well understood. In humans and mice, epithelial cell-derived thymic stromal lymphopoietin (TSLP) plays a central role in this process. Little is known about equine TSLP (eqTSLP) and its role in allergic diseases and our current knowledge is limited to the assessment of TSLP mRNA expression. In order to be able to study eqTSLP at the protein level, the aim of the present study was to produce recombinant eqTSLP protein and generate TSLP-specific antibodies. EqTSLP was cloned from a skin biopsy sample from a horse with chronic urticaria and eqTSLP protein was expressed in E.coli and in mammalian cells. Recombinant proteins were designed to include C-terminal Histag, which allowed subsequent purification and detection by Histag-specific Ab. Polyclonal and monoclonal eqTSLP-specific Ab were generated after immunization of mice with E.coli-expressed TSLP. Their specificity was tested by western blotting and ELISA. In addition, a commercially available polyclonal human TSLP-specific antibody was tested for cross-reactivity with eqTSLP. Expression of TSLP protein was confirmed by western blotting using Histag-specific Ab. E.coli-expressed TSLP appears as a band of ～13 kDa, whereas mammalian cell-expressed TSLP showed several bands at 20-25 kDa, probably representing several glycosylation forms. Polyclonal and monoclonal antibodies generated in this study, as well as commercially available human TSLP-specific Ab reacted with both E.coli- and mammalian cell-expressed TSLP in western blotting and ELISA. A capture ELISA was established to quantitate TSLP in cell supernatants and validated using supernatants from primary equine keratinocytes and peripheral blood leukocytes (PBL). Increased TSLP concentrations were found after stimulation of keratinocytes with PMA+ionomycine and with Culicoides extract. Similarly, increased TSLP concentrations were detected in PBL after stimulation with ConA, Culicoides extract, or IgE cross-linking. In conclusion, recombinant TSLP proteins and TSLP-specific antibodies produced in this study will allow further studies of the role of TSLP in equine allergic diseases.     

PIN proteins perform a rate-limiting function in cellular auxin efflux

Intercellular flow of the phytohormone auxin underpins multiple developmental processes in plants. Plant-specific pin-formed (PIN) proteins and several phosphoglycoprotein (PGP) transporters are crucial factors in auxin transport-related development, yet the molecular function of PINs remains unknown. Here, we show that PINs mediate auxin efflux from mammalian and yeast cells without needing additional plant-specific factors. Conditional gain-of-function alleles and quantitative measurements of auxin accumulation in Arabidopsis and tobacco cultured cells revealed that the action of PINs in auxin efflux is distinct from PGP, rate-limiting, specific to auxins, and sensitive to auxin transport inhibitors. This suggests a direct involvement of PINs in catalyzing cellular auxin efflux.     

Effect of Azospirillum inoculants on arbuscular mycorrhiza establishment in wheat and maize plants

Plant growth-promoting rhizobacteria and arbuscular mycorrhizal (AM) fungi represent two main groups of beneficial microorganisms of the rhizosphere. The role of different strains of Azospirillum on AM fungi development was evaluated by measuring the percentage of AM colonisation of the root system in durum wheat and maize plants, grown under both greenhouse and field conditions. The effect of wild-type Azospirillum brasilense strain Sp245 and genetically modified (GM) derivatives overproducing indole-3-acetic acid was assessed at greenhouse level in (1) three different cultivars of durum wheat, in the presence of indigenous AM fungi and (2) maize plants artificially inoculated with Glomus mosseae and Glomus macrocarpum. In addition, the establishment of natural AM fungal symbiosis was evaluated using Azospirillum lipoferum CRT1 in maize plants at field level. Despite the stimulatory effect of the different Azospirillum inocula on root growth, no significant differences in AM colonisation were found, independently of the AM fungus involved, either in wheat or in maize plants. Similarly, GM A. brasilense, which strongly stimulates root development, did not affect AM formation. Although these results were obtained in conditions in which the mycorrhization rate was moderate (15–30%), overall considered they indicate that the use of wild-type or GM Azospirillum phytostimulators does not alter mycorrhization.     

Immunochemical approach to detection of adulteration in honey: physiologically active royal jelly protein stimulating TNF-alpha release is a regular component of honey

The presence of royal jelly (RJ) proteins in honey collected from nectars of different plants, origin, and regions and in honeybee's pollen was detected by Western-blot analysis using polyclonal antibodies raised against water-soluble RJ-proteins. The most abundant RJ-protein in honeybee products corresponded to a 55 kDa protein. The N-terminal amino acid sequence of 55 kDa protein was N-I-L-R-G-E. This sequence is identical to the apalbumin-1, the most abundant protein of RJ. Apalbumin-1 is a regular component of honeybee products and thus is a suitable marker tool for proving adulteration of honey by means of immunochemical detection. Its presence in all tested samples of honeys and honeybee pollen was confirmed also by Western-blot analysis using polyclonal antibodies raised against recombinant apalbumin-1. It has been found that major RJ-proteins, apalbumin-1, and apalbumin-2, stimulate mouse macrophages to release TNF-alpha, which demonstrates that physiologically active proteins of honey could be used for its biological valuation.     

Stone bunds for soil conservation in the northern Ethiopian highlands: Impacts on soil fertility and crop yield

In the Ethiopian highlands, large-scale stone bund building programs are implemented to curb severe soil erosion. Development of soil fertility gradients is often mentioned as the major drawback of stone bund implementation, as it would result in a dramatic lowering of crop yield. Therefore, the objectives of this study are to assess soil fertility gradients on progressive terraces and their influence on crop yield, in order to evaluate the long-term sustainability of stone bunds in the Ethiopian Highlands.

The study was performed near Hagere Selam, Tigray and comprises (i) measurement of P_av, N_tot and C_org along the slope on 20 representative plots and (ii) crop response measurement on 143 plots. Results indicate that levels of P_av, N_tot and C_org in the plough layer are highly variable between plots and mainly determined by small-scale soil and environmental features, plot history and management. After correcting for this “plot effect” a significant relationship (p < 0.01) was found between the position in the plot relative to the stone bund and levels of P_av and N_tot, which are higher near the lower stone bund, especially on limestone parent material. For C_org and on basalt-derived soils in general no significant relationship was found. Although soil fertility gradients are present, they are not problematic and can be compensated by adapted soil management. Only in areas where a Calcaric or Calcic horizon is present at shallow depth, care should be taken. Crop Yields increased by 7% compared to the situation without stone bunds, if a land occupation of 8% by the structures is accounted for. Yield increased from 632 to 683 kg ha⁻¹ for cereals, from 501 to 556 kg ha⁻¹ (11%) for Eragrostis tef and from 335 to 351 kg ha⁻¹ for Cicer arietinum.

No negative effects reducing stone-bund sustainability were found in this study. Soil erosion on the other hand, poses a major threat to agricultural productivity. Stone bund implementation therefore is of vital importance in fighting desertification and establishing sustainable agriculture in the Ethiopian highlands.     

Toward a kinetic model for acrylamide formation in a glucose-asparagine reaction system

A kinetic model for the formation of acrylamide in a glucose-asparagine reaction system is pro-posed. Equimolar solutions (0.2 M) of glucose and asparagine were heated at different temperatures (120-200 degrees C) at pH 6.8. Besides the reactants, acrylamide, fructose, and melanoidins were quantified after predetermined heating times (0-45 min). Multiresponse modeling by use of nonlinear regression with the determinant criterion was used to estimate model parameters. The proposed model resulted in a reasonable estimation for the formation of acrylamide in an aqueous model system, although the behavior of glucose, fructose, and asparagine was slightly underestimated. The formation of acrylamide reached its maximum when the concentration of sugars was reduced to about 0. This supported previous research, showing that a carbonyl source is needed for the formation of acrylamide from asparagine. Furthermore, it is observed that acrylamide is an intermediate of the Maillard reaction rather than an end product, which implies that it is also subject to a degradation reaction.     

A 12-month survey of gastrointestinal helminth infections of cervids kept in two zoos in Belgium.

Infections with helminths are a major health issue in captive and wild deer. In this study, fecal egg count patterns and clinical signs associated with gastrointestinal nematodes were assessed for 12 mo in nine cervid herds kept under different husbandry conditions at two sites. At site 1, an urban zoo, fecal egg counts remained low and no clinical signs of parasitic gastroenteritis were seen in the herds of fallow deer (Dama dama), Dybowski's deer (Cervus nippon dybowski), pudu (Pudu pudu), and reindeer (Rangifer tarandus tarandus). Helminth infection at this site may have been successfully prevented by daily dung removal of the small sandy-soil enclosures, and applying routine anthelmintic treatment was not justified. At site 2, a wild animal park, involved species were red deer (Cervus elaphus hippelaphus), Nelson's elk (Cervus elaphus nelsoni), Père David's deer (Elaphurus davidianus), European elk (Alces alces alces), and reindeer (Rangifer tarandus tarandus). Nematode eggs were frequently encountered in herds of red deer, Nelson's elk, and European elk, which were kept on larger, grassy enclosures that were irregularly cleaned. The trimodal pattern of fecal egg counts in herds from the wild animal park, consisting of a small spring rise in June, a peak in October, and a small rise in February, indicates that infective larvae on pastures are the main source of infection. In addition, routine anthelmintic treatment with fenbendazole in April and July limited egg shedding, but reinfection rapidly occurred. In two European elk and one reindeer, increasing fecal egg counts were associated with loss of fecal consistency and reduced appetite. Three genera and three species of nematodes were recovered at necropsy of one red deer and three Nelson's elk: Spiculopteragia spiculoptera, Trichostrongylus spp., Nematodirus filicollis, Capillaria spp., Oesophagostomum radiatum, and Trichuris spp., with total worm counts between 950 and 8,700.