Novel antigens for detection of cell mediated immune responses to Mycobacterium avium subsp. paratuberculosis infection in cattle

Paratuberculosis is a chronic infection of the intestine of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Early stage MAP infection can be detected by measuring specific cell mediated immune responses, using the whole blood interferon-γ (IFN-γ) assay. Available IFN-γ assays use purified protein derivative of MAP (PPDj) which are complex antigen mixtures with low specificity. The objectives of this study were to evaluate immunogenicity and specificity of 14 novel recombinant antigens for use in the IFN-γ assay and to assess the consistency of IFN-γ responses. The study included blood samples from 26 heifers from a MAP infected herd, collected three times with four to five-week intervals, and blood samples from 60 heifers of a non-infected herd collected once. Heifers of the non-infected herd were used to establish cut-off values for each antigen. The case definition was an animal with ≥ 2 positive tests for ≥ 4 antigens, resulting in 13 cases and 13 non-cases. IFN-γ levels of cases were higher compared to IFN-γ levels of non-cases (P<0.05). The results of the IFN-γ assay using PPDj did not correlate well with the results using the novel antigens. PPDj produced elevated IFN-γ responses of samples from both the non-infected and the MAP infected herd, indicating unspecific IFN-γ responses and showed low consistency. Three latency proteins, LATP-1, LATP-2 and LATP-3 gave positive IFN-γ tests that correlated very well with the case definition suggesting high immunogenicity. Three tested antigens, LATP-2, MAP-1 and MAP-2 have no homologue in the M. avium subsp. avium or M. bovis genome and could be promising diagnostic antigens, especially LATP-2 correlated highly with the case definition.     

Effects of elevated atmospheric CO2, prolonged summer drought and temperature increase on N2O and CH4 fluxes in a temperate heathland

In temperate regions, climate change is predicted to increase annual mean temperature and intensify the duration and frequency of summer droughts, which together with elevated atmospheric carbon dioxide (CO₂) concentrations, may affect the exchange of nitrous oxide (N₂O) and methane (CH₄) between terrestrial ecosystems and the atmosphere. We report results from the CLIMAITE experiment, where the effects of these three climate change parameters were investigated solely and in all combinations in a temperate heathland. Field measurements of N₂O and CH₄ fluxes took place 1-2 years after the climate change manipulations were initiated. The soil was generally a net sink for atmospheric CH₄. Elevated temperature (T) increased the CH₄ uptake by on average 10 μg C m⁻² h⁻¹, corresponding to a rise in the uptake rate of about 20%. However, during winter elevated CO₂ (CO₂) reduced the CH₄ uptake, which outweighed the positive effect of warming when analyzed across the study period. Emissions of N₂O were generally low (<10 μg N m⁻² h⁻¹). As single experimental factors, elevated CO₂, temperature and summer drought (D) had no major effect on the N₂O fluxes, but the combination of CO₂ and warming (TCO₂) stimulated N₂O emission, whereas the N₂O emission ceased when CO₂ was combined with drought (DCO₂). We suggest that these N₂O responses are related to increased rhizodeposition under elevated CO₂ combined with increased and reduced nitrogen turnover rates caused by warming and drought, respectively. The N₂O flux in the multifactor treatment TDCO₂ was not different from the ambient control treatment. Overall, our study suggests that in the future, CH₄ uptake may increase slightly, while N₂O emission will remain unchanged in temperate ecosystems on well-aerated soils. However, we propose that continued exposure to altered climate could potentially change the greenhouse gas flux pattern in the investigated heathland.     

Xanthohumol uptake and intracellular kinetics in hepatocytes, hepatic stellate cells, and intestinal cells

Xanthohumol (XN) is the major prenylated chalcone of hops and hence an ingredient of beer. Despite many advances in understanding of the pharmacology of XN, one largely unresolved issue is its low bioavailability in the human organism. Also, not much is known about its actual concentrations and pharmacokinetics in liver and intestinal cells. Therefore, the uptake, intracellular distribution, and kinetics of XN were studied in various cell types, namely, hepatocellular carcinoma cells (HuH-7), hepatic stellate cells (HSC), primary cultured hepatocytes, and colorectal adenocarcinoma cells (Caco-2). Fluorescent microscopy allowed for the first time visualization and tracing of the uptake and intracellular distribution of XN. A rapid accumulation of XN concentrations that were up to >60-fold higher than the concentration present in the ambient culture medium was observed. Fluorescence recovery after photobleaching experiments revealed that most XN molecules are bound to cellular proteins, which may alter properties of cellular factors.     

Anemia and Growth Retardation in Totally Gastrectomized Swine

The effects of total gastrectomy in six young swine were followed and described during an experimental period up to 18 months. Two of the gastrectomized pigs (Nos. 1 and 2) were medicated with cycobemin every second week. Two other gastrectomized pigs (Nos. 11 and 14) were medicated with iron-dextran twice a week. Three non-gastrectomized swine receiving the same feed as the gastrectomized animals were observed as controls for up to eight months.One of the gastrectomized non-medicated pigs (No. 4) did not gain weight, while the other gastrectomized animals had a normal appearance and gained weight but less than the controls. The non-medicated as well as the cycobemin-medicated gastrectomized animals developed a microcytic hypochrome anemia corresponding to the anemia in iron deficiency. Histochemically loss of iron in the depots (liver, spleen, bone marrow and intestine) was demonstrated in the gastrectomized animals except those treated with iron-dextran. Histological and histochemical examinations of the nervous system did not show any abnormalities.     

Effects of feeding fat-coated butyrate on mucosal morphology and function in the small intestine of the pig

As shown earlier, pig rations with high starch and purine content initiate mucosal hypertrophy by stimulating mitotic activity and DNA formation in the small intestine, whereas in the colon butyrate inhibits apoptosis and thus increases crypt depth. It was the aim of this study to combine these effects by targeting fat-coated butyrate into the small intestine where it usually does not occur, and to investigate effects on mucosal development and function. Three groups of five pigs were fed 3.6 kg/day of either a low-energy ration [deficit group, 6.6 MJ metabolizable energy (ME)/kg] or a high-energy ration (13.7 MJ ME/kg) that was supplemented with brewing yeast as a source of purines. The third ration was of high energy and contained purines and was additionally supplemented with coated butyrate (13.5 MJ ME/kg; 29 g calcium butyrate/kg). Rations were fed for 5 days. After killing, tissue samples were obtained from the proximal, medial and distal parts of jejunum for histology. Chyme samples were obtained from the ileum of all animals and used for sucrase determination. Villus size was not changed by feeding, but butyrate had an effect on plica height and area mainly in the medial jejunum. Plica area in the butyrate group (4.2 mm(2)) was significantly higher (p 相似文献   

Prevention strategies for field traffic-induced subsoil compaction: a review: Part 2. Equipment and field practices

The loads imposed by modern farm machinery have considerable potential to increase subsoil stress. Within the context of economically viable and environmentally sustainable systems, the practices associated with subsoil damage and methods for avoidance are identified. The greatest potential for damage is on fragile, wet or loosened subsoils combined with high wheel or track loads and contact pressures that create noticeable ruts in the topsoil. In-furrow ploughing increases this potential considerably by placing loads on the subsoil. Measures to avoid this potential involve a whole farm approach and an understanding of the many interactions between cropping systems and machinery. Alternatives to in-furrow ploughing that involve working from the surface and building a protective topsoil are discussed. Key measures to reduce the risk to subsoils involve a clear understanding of tyre load and inflation data and simple on-farm methods of achieving this are suggested. Although avoidance has the potential to reduce the risk, confinement of damage to specific strips in the field is seen as a realistic alternative. Controlled traffic operations, together with precision guidance, offer an economic means by which compaction on the cropped area can be avoided. The most effective route to improvement in soil care across the European Union (EU) is an appropriate management structure coupled with a best practice framework.     

A catena in northeast Brandenburg (Bölkendorf): Soil biota,root growth and soil structure

Typical for the morainal landscape of NE‐Germany are catenas with pronounced gradients of abiotic factors, such as soil texture and organic matter content. For adequate management of these regions, their ecology must be better understood. In this context, the biological interactions in soil play an important role. The present study is based on the hypothesis that the gradients of abiotic factors across the catena are reflected in soil biota, i.e. earthworms and microarthropods, and root growth.

Sampling was carried out at selected positions across the catenary sequence of Boelkendorf, Brandenburg, for earthworms, Collembola and roots in April 1994. The soil physical analysis comprised measurements of soil bulk density and penetration resistance.

A positive correlation between earthworm abundance, numbers of Collembola and root growth at different catena positions was found. The top position especially, a calcaric regosol with low organic matter content, was markedly different with respect to soil biological and physical characteristics. High bulk densities and high soil resistance in the subsoil coincided with low earthworm densities, low microarthropod numbers and impeded rootability of the soil.     

Normal electrocardiographic QT interval in race-fit Standardbred horses at rest and its rate dependence during exercise

ObjectivesCardiac repolarization, measured as QT and T_peak to T_end (T_pT_e) intervals on the ECG, is important, as irregularities caused by diseases, ventricular hypertrophy, drugs and genetic defects can trigger arrhythmias which predispose human patients to syncope and sudden cardiac death. In horses, repolarization is not well described and therefore QT analysis cannot yet be used diagnostically. Therefore, we sought to describe reference values for the normal QT and T_pT_e intervals in Standardbreds and to determine the best method for heart rate (HR) correction.Animals30 Standardbreds.MethodsQT and T_pT_e intervals were measured during rest and exercise and plotted against HR converted to R_peak to R_peak interval (RR). Data were fitted with relevant regression models. Intra- and inter-observer agreement was assessed using Bland–Altman analyses.ResultsData were best described by a piecewise linear model (r² > 0.97). Average prediction error of this model was smaller than for both Bazett and Fridericia corrections. Coefficient of repeatability of intra- and inter-observer variability was 8.76 ms and 5.64 ms respectively and coefficient of variation was 1.77% and 2.76% respectively. T_pT_e increased with RR in stallions.ConclusionsThe QT interval in Standardbred horses shortens with decreasing RR interval (increasing HR) as in humans, but in a markedly different order as it clearly follows a piecewise linear model. The equine QT interval can be measured easily and there is small intra- and inter-observer variability. This model of the equine QT interval provides clinicians with a method that could support a diagnosis of repolarization disturbances in horses.