Competitive enzyme-linked immunosorbent assay for heartwater using monoclonal antibodies to a Cowdria ruminantium-specific 32-kilodalton protein

Hybridomas producing monoclonal antibodies (mAb) to Cowdria ruminantium were raised. Four mAbs of the IgG isotype reacted in western blots with a 32-kilodalton Cowdria protein (Cr32), which had previously been shown to be conserved and immunodominant. A fifth mAb of the IgM isotype recognized a 40-kDa Cowdria protein. The latter mAb was negative in an indirect fluorescent antibody test (IFA), whereas the other four were positive. mAb No. 4F10B4 showed the strongest signal in western blots using three different stocks of Cowdria. Immuno-gold labeling of Cowdria organisms in vitro using 4F10B4 showed that Cr32 has surface-exposed antigenic determinants. Using mAb 4F10B4, a competitive ELISA was developed which detected specific Cowdria antibodies in goat, sheep and cattle sera. Antibodies in animal sera competed with binding of mAb 4F10B4 to a crude sonicated Cowdria antigen obtained from infected endothelial cell cultures. The competition ELISA (CELISA) detected antibodies in 55 out of 70 (79%) goats experimentally infected with one of eight different Cowdria stocks. Fourteen out of the 15 sera which were shown negative in the CELISA were also negative in the IFA. Nevertheless, all 15 sera recognized some epitopes of the immunodominant Cowdria-specific 32 kDa protein as judged from their reaction with this protein in western blots. Overall, there was 89% agreement between CELISA and IFA considering all 70 goat sera. Moreover, antibodies were detected in nine out of nine sheep infected with one of three different stocks of Cowdria and in sera from calves experimentally infected by two different strains of heartwater. There were no cross-reactions with Ehrlichia phagocytophila antibodies in goat sera, nor with Anaplasma marginale antibodies in bovine sera. Lack of cross-reactivity and detection of antibodies to eight geographically widely distributed stocks of Cowdria, makes the competition ELISA a promising test for use in heartwater endemic areas.     

Schmallenberg virus epidemic: Impact on milk production,reproductive performance and mortality in dairy cattle in the Netherlands and Kleve district,Germany

Schmallenberg virus (SBV), a novel orthobunyavirus that rapidly spread throughout north-western Europe in 2011, caused congenital malformations in lambs and goat kids (Van den Brom et al., 2012) and newborn calves (Hoffmann et al., 2012). The impact of the SBV epidemic seemed limited however, in terms of the number of affected herds with malformed offspring (European Food Safety Authority, 2012b). Nevertheless, little is known with regard to the overall within-herd impact of SBV infection. The objective of the current study was to quantify the impact of the 2011 SBV epidemic on the productivity of dairy cattle in the Netherlands and the district of Kleve, Germany.     

Adherence of Actinobacillus pleuropneumoniae to primary cultures of porcine lung epithelial cells

To study adherence of Actinobacillus pleuropneumoniae to porcine lower respiratory epithelium, a cell culture model was developed using primary cultures of porcine lung epithelial cells (LEC). Adherence assays were performed and results were compared with data obtained with swine kidney cells (SK6). A. pleuropneumoniae efficiently adhered to LEC with up to 62 bacteria per cell after 2h of incubation. Reference strain of serotype 3 (R3) adhered better to LEC than reference strains of serotypes 1 (R1), 7 (R7) and 8 (R8). Overall the adherence to LEC was more rapid and up to 30-fold more efficient than adherence to SK6 cells. In search for the mechanism involved in the adherence event, we tested the effect of LPS which has previously been demonstrated to cause adherence of the pathogen to upper respiratory epithelium. Adherence assays with LPS transposon mutants demonstrated unaltered (mutant with modification in core/lipid A moiety) or even three-fold more adherence (mutants lacking O antigen) compared to the parent micro-organisms. Purified LPS of strains R1, R3, R7 and R8 did not inhibit adherence of R8 to LEC either, suggesting that LPS and particularly the O-antigen are not essential for adherence of A. pleuropneumoniae to LEC. The efficient, LPS-independent adherence of A. pleuropneumoniae to LEC cells indicates that A. pleuropneumoniae may carry different, cell type-specific adhesins and that primary cultures of lower respiratory epithelium are valuable infection models in studying A. pleuropneumoniae pathogenesis.     

Back kinematics of healthy trotting horses during treadmill versus over ground locomotion

Reasons for performing study: Treadmill locomotion is frequently used for training of sport horses, for diagnostic purposes and for research. Identification of the possible biomechanical differences and similarities between the back movement during treadmill (T) and over ground (O) locomotion is essential for the correct interpretation of research results. Objectives: To compare the kinematics of the thoracolumbar vertebral column in treadmill and over ground locomotion in healthy horses. Methods: Six sound Dutch Warmblood horses trotted on a T and O during 10 s at their own preferred velocity (mean ± s.d. 3.6 ± 0.3 m/s T and 3.6 ± 0.1 m/s O), which was the same in both conditions. Kinematics of the vertebral column was captured by infrared cameras using reflective skin markers attached over the spinous processes of selected vertebrae and other locations. Flexion‐extension and lateral bending range of motion (ROM), angular motion pattern (AMP) and intravertebral pattern symmetry (IVPS) of 5 vertebral angles (T6‐T10‐T13, T10‐T13‐T17, T13‐T17‐L1, T17‐L1‐L3 and L1‐L3‐l5) were calculated. Neck angle, linear and temporal stride parameters and protraction‐retraction angles of the limbs were also calculated. Results: The vertical ROM (flexion‐extension) was similar in both conditions, but the horizontal ROM (lateral bending) of the lumbar angles T17‐L1‐L3 and L1‐L3‐L5 was less during T locomotion (mean ± s.d. difference of 1.8 ± 0.6 and 1.7 ± 0.9°, respectively, P>0.05). During O locomotion, the symmetry pattern of the lumbar vertebral angles was diminished from 0.9 to 0.7 (1 = 100% symmetry) indicating increased irregularity of the movement (P>0.05). No differences were found in the basic linear and temporal stride parameters and neck angle. Potential relevance: Vertebral kinematics during treadmill locomotion is not identical to over ground locomotion, but the differences are minor. During treadmill locomotion lumbar motion is less, and caution should be therefore taken when interpreting lumbar kinematics.     

A Single Sample Method for Evaluating 51Chromium-Ethylene Diaminic Tetraacetic Acid Clearance in Normal and Hyperthyroid Cats

Background: Chronic kidney failure is frequently seen in middle-aged and elderly cats. ⁵¹Chromium-ethylene diaminic tetraacetic acid (⁵¹Cr-EDTA) clearance and single blood sample (SBS) method are used in several species to estimate the glomerular filtration rate (GFR).
Hypothesis: The hypothesis of this study was that ⁵¹Cr-EDTA clearance could be determined using an SBS method in normal and hyperthyroid cats.
Animals: Forty-six cats were included in this study, with an average age of 9.5 years. Of these cats, 27 had hyperthyroidism; 19 were healthy.
Methods: After IV injection of ⁵¹Cr-EDTA (average dose: 4.25 MBq), 7 blood samples were obtained between 5 and 240 minutes. Reference clearance was calculated in mL/min and mL/min/kg body weight, using a 2-compartment model. Optimal time for clearance measurement with SBS was then determined by systematically comparing each individual plasma concentration to the reference multisample clearance.
Results: The average reference plasma clearance of ⁵¹Cr-EDTA for all cats was 14.9 mL/min (3.7 mL/min/kg). The clearance in hyperthyroid cats averaged 16.4 mL/min (4.3 mL/min/kg) and in normal cats averaged 10.3 mL/min (2.4 mL/min/kg).
The optimal time for the SBS was 48 minutes after injection of tracer ⁵¹Cr-EDTA ( R ²= 0.9414), giving the following converting equation: clearance = (0.0066 × DV_{48 minutes}) – 0.9277 (in mL/min).
Conclusions and Clinical Importance: In this study, the single sample ⁵¹Cr-EDTA clearance method was used to estimate the global GFR in cats. The method identified differences in clearance between normal and hyperthyroid cats. The optimal time for an SBS was 48 minutes.     

Crystal structure of the long-chain fatty acid transporter FadL

The mechanisms by which hydrophobic molecules, such as long-chain fatty acids, enter cells are poorly understood. In Gram-negative bacteria, the lipopolysaccharide layer in the outer membrane is an efficient barrier for fatty acids and aromatic hydrocarbons destined for biodegradation. We report crystal structures of the long-chain fatty acid transporter FadL from Escherichia coli at 2.6 and 2.8 angstrom resolution. FadL forms a 14-stranded beta barrel that is occluded by a central hatch domain. The structures suggest that hydrophobic compounds bind to multiple sites in FadL and use a transport mechanism that involves spontaneous conformational changes in the hatch.     

Submarine thermal sprirngs on the galapagos rift

The submarine hydrothermal activity on and near the Galápagos Rift has been explored with the aid of the deep submersible Alvin. Analyses of water samples from hydrothermal vents reveal that hydrothermal activity provides significant or dominant sources and sinks for several components of seawater; studies of conductive and convective heat transfer suggest that two-thirds of the heat lost from new oceanic lithosphere at the Galápagos Rift in the first million years may be vented from thermal springs, predominantly along the axial ridge within the rift valley. The vent areas are populated by animal communities. They appear to utilize chemosynthesis by sulfur-oxidizing bacteria to derive their entire energy supply from reactions between the seawater and the rocks at high temperatures, rather than photosynthesis.     

Analysing agricultural users’ patterns of behaviour: The case of OPTIRas™, a decision support system for starch crop selection

Redesigning IT systems for specific user groups encompasses a lot of effort with respect to analysing and understanding user behaviour. The goal of this paper is to provide insights into patterns of behaviour of agricultural users, during the usage of a decision support system called OPTIRas™. This system aids agricultural users in their cultivar selection activities. We analyse logs resulting from OPTIRas™, and we get insights into user’s navigational patterns. We claim that the results of our analysis can be used to support the redesign of decision support systems in order to address specific agricultural users’ characteristics.     

Effects on nutrients and on grain quality in spring wheat crops grown under elevated CO2 concentrations and stress conditions in the European, multiple-site experiment ‘ESPACE-wheat’

Nutrient element concentrations and grain quality were assessed in spring wheat grown under elevated CO₂ concentrations and contrasting levels of tropospheric ozone at different nitrogen supply rates at several European sites. Carbon dioxide enrichment proved to affect nutrient concentrations in a complex manner. In green leaves, all elements (with exception of phosphorus and iron) decreased. In contrast, effects on the element composition of grains were restricted to reductions in nitrogen, calcium, sulphur and iron. Ozone exposure resulted in no significant effects on nutrient element concentrations in different tissues in the overall analysis. The nitrogen demand of green tissues was reduced due to CO₂ enrichment as shown by reductions in the critical leaf nitrogen concentration and also enhanced nitrogen use efficiency. Reductions in the content of ribulose-bisphosphate carboxylase/oxygenase and repression of the photorespiratory pathway and reduced nitrogen allocation to enzymes driving the photosynthetic carbon oxidation cycle were chiefly responsible for this effect. Thus, nitrogen acquisition by the crop did not match carbon acquisition under CO₂ enrichment. Since crop nitrogen uptake from the soil was already completed at anthesis, nitrogen allocated to the grain after anthesis originated from vegetative pools—causing grain nitrogen concentrations to decrease under CO₂ enrichment (on average by 15% when CO₂ concentrations increased from 360 to 680 μmol mol⁻¹). Correspondingly, grain quality was reduced by CO₂ enrichment. The Zeleny value, Hagberg value and dry/wet gluten content decreased significantly with increasing [CO₂]. Despite the beneficial impact of CO₂ enrichment on growth and yield of C₃ cereal crops, declines in flour quality due to reduced nitrogen content are likely in a future, [CO₂]-rich world.