Evaluation of the Accuracy of Diagnostic Tests From Repeated Measurements Without a Gold Standard

A model is presented to evaluate the accuracy of diagnostic tests from data from individuals that are repeatedly tested in time. Repeated measurements from three diagnostic tests for foot-and-mouth disease, applied to vaccinated and experimentally infected cattle, were analyzed. At any time the true disease status of the individuals was unknown, i.e., no gold standard was available. The model allows for correlation between repeated test results, in consequence of the underlying structure for the unknown true disease status, but also by the distribution of the test results conditional upon true disease status. The model also allows for dependence between the different diagnostic tests conditional upon true disease status. Prior information about the structure of the prevalence and the specificity of the tests was incorporated in a Bayesian analysis. Posterior inference was carried out with Markov chain Monte Carlo. Simulated data were analyzed to gain insight into the performance of the posterior Bayesian inference. The simulated data are typical for the expensive and, therefore, modestly sized infection experiments that are conducted under controlled conditions.     

Immune responses in Eimeria acervulina infected one-day-old broilers compared to amount of Eimeria in the duodenum, measured by real-time PCR

T-cell responses are supposed to be the major immune reactions in broilers infected with Eimeria. The nature of such T-cell responses is influenced by the species of Eimeria involved, age of the host, amount of parasites and the preceding infection history. In young chicks the intestine is still developing in length while the lymphocyte populations in the gut develop and differentiate. In chicks infected at young age the immune response may be different in quality as compared to responses in adults. We investigated the (T-cell) immune responses of young broilers to a primary Eimeria acervulina infection in relation to the number of parasites used for infection. In our experiment we infected one-day-old broilers with a low (5 x 10(2) oocysts) and a high (5 x 10(4) oocysts) dose of E. acervulina. We used a newly developed species specific real-time PCR to quantify total amount of parasites in the duodenum as the number of oocysts in faeces may not be representative for the exposure of the gut immune system. We characterized T-cell subsets in the duodenum by means of FACS-analyses, lymphocyte proliferation assays with spleen lymphocytes and the mRNA profiles of different cytokines (TGF-beta2, -4, IFN-gamma, IL-2, -6, -8 and -18) in the duodenum by means of real-time PCR. From day 5 p.i. broilers with a high dose of E. acervulina had a significantly lower body weight than the control group. No increase in CD4(+) cells, but a strong increase in CD8(+) cells was observed at days 7 and 9 p.i. in the duodenum of broilers infected with a high dose E. acervulina. IL-8 mRNA responses were observed after infection with low and with high infection doses, but no IFN-gamma and TGF-beta mRNA responses were found in the duodenum. The specific proliferative T-cell responses to a low infectious dose were not significantly different as compared to the control group. In conclusion, based on the kinetics of observed responses a primary infection with a high dose of E. acervulina in one-day-old broilers seems to generate an immune response that shows a peak at the time of oocyst excretion, whereas the immune response to a low dose is less explicit.     

Multiple segmented reaches per subwatershed modeling approach for improving HSPF-Paddy water quality simulation

Reach segmentation influences predicted water quality concentrations in water quality modeling. Many lumped or semi-distributed watershed models, including Soil and Water Assessment Tool (SWAT) and Hydrologic Simulation Program-Fortran (HSPF), simulate stream/river water quality with a single segmented reach per subwatershed (SSRS) modeling approach. A multiple-segmented reaches per subwatershed (MSRS) modeling approach was developed, and simulated BOD₅ concentrations for this approach were compared with results from the SSRS modeling approach using the HSPF-Paddy model. The SSRS modeling approach has potential systemic errors for predicting BOD₅ concentration even when the model is well calibrated. When the point source was loaded at the most upstream location with the same decay rate, the predicted BOD₅ concentration using the SSRS modeling approach was higher compared with results for the MSRS modeling approach, and the difference between two methods increases with increasing load and decay rate. When BOD₅ was loaded more downstream, BOD₅ concentration using the SSRS modeling approach was lower compared with results for MSRS modeling. For a case study, simulated streamflow and BOD₅ concentration for the SSRS and MSRS modeling approaches demonstrated good agreement with observed data. However, the estimated decay rate for the SSRS modeling approach was smaller than that for the MSRS modeling approach because BOD decays through total volume in the SSRS modeling approach, although BOD may be loaded anywhere in the reach. The MSRS modeling approach can minimize systematic errors and provide more detailed variation of water quality concentrations along a river length. The MSRS modeling approach does not always need to be applied to all subwatersheds but is recommended for reaches significantly polluted by point source pollution.     

The Influence of a Particle Separation Technology on the Generation of Airborne Particles from Different Roughages and Bedding Materials Used for Horses

Four different types of bedding materials (wheat straw, wood shavings, hemp shives, flax shives) and two roughages (hay, haylage) were treated using an air-driven particle separation technology. The airborne particle and mold generation of both treated and untreated samples were then analyzed under standardized laboratory conditions. In addition, samples of all the treated materials were stored for 8 weeks either in a pressed or incoherent form and then analyzed again for their ability to generate airborne particles. The airborne particle concentrations were detected online with the gravimetrically measuring analyzer tapered element oscillating microbalance (TEOM) 1400a that was equipped successively with different inlets to measure the particle fractions PM₂₀, PM₁₀, PM_2.5, and PM_1.0 (PM = Particulate matter). The particle separation resulted in a reduction in the airborne particle (PM₂₀) generation in all materials: hay 49.16 to 22.79 mg/m³ (53.6%), haylage 28.57 to 25.04 mg/m³ (12.3%), wood shavings 141.68 to 15.04 mg/m³ (89.4%), wheat straw 143.08 to 22.97 mg/m³ (83.9%), flax 135.11 to 53.31 mg/m³ (60.5%), and hemp 63.67 to 17.64 mg/m³ (72.3%). The 8-week storage of the treated materials as compressed materials led to a renewed significant increase in the airborne particle (PM₁₀) concentration in the haylage (+29.9%), wheat straw (+104.0%), wood shavings (+40.4%), and hemp shives (+30.7%). Storage of the incoherent materials caused a significant increase in these particles only in the wheat straw (+44.2%). The separation treatment reduced the mold production by 92.4% in the wood shavings, 88.0% in the wheat straw, and 85.8% in the hay.     

Simultaneous analysis of free phytosterols/phytostanols and intact phytosteryl/phytostanyl fatty acid and phenolic acid esters in cereals

An approach based on solid-phase extraction for the effective separation of free phytosterols/phytostanols and phytosteryl/phytostanyl fatty acid and phenolic acid esters from cereal lipids was developed. The ester conjugates were analyzed in their intact form by means of capillary gas chromatography. Besides free sterols and stanols, up to 33 different fatty acid and phenolic acid esters were identified in four different cereal grains via gas chromatography-mass spectrometry. The majority (52-57%) of the sterols and stanols were present as fatty acid esters. The highest levels of all three sterol and stanol classes based on dry matter of ground kernels were determined in corn, whereas the oil extract of rye was 1.7 and 1.6 times richer in fatty acid esters and free sterols/stanols than the corn oil. The results showed that there are considerable differences in the sterols/stanols and their ester profiles and contents obtained from corn compared to rye, wheat, and spelt. The proposed method is useful for the quantification of a wide range of free phytosterols/phytostanols and intact phytosteryl/phytostanyl esters to characterize different types of grain.     

Temporary Turnout for Free Exercise in Groups: Effects on the Behavior of Competition Horses Housed in Single Stalls

In Germany, most competition horses are housed in single stalls and free exercise is not permitted in many cases. The reason for not allowing free exercise is mostly the risk of injury. Additionally, opinions exist that the horses’ demand for exercise is fulfilled by training and that the horses’ willingness to perform is negatively influenced by free exercise. In the present study, three turnout practices were investigated with regard to their effect on the behavior of four horses: daily training without free exercise (no turnout), 2-hour turnout (for free exercise) before training, and 2-hour turnout after training. The aim of this study was to determine any differences in the horses’ behavior between the three treatments. The horses’ behavior in the stable was observed through video recordings. The behavior during turnout was observed directly and during training was evaluated by the riders with the aid of a questionnaire. Additionally, the distance covered during turnout was measured by Global Positioning System devices. The behavior within the stall was more restless in the treatment without turnout—which became apparent in significantly more frequent changes between behaviors as compared with the treatments with turnout. The results of Global Positioning System measurement during turnout showed a significantly shorter distance covered when horses were trained before they were turned out compared with turnout before training. If the horses were turned out after training, they also showed less trotting and cantering and more dozing. The horses’ willingness to perform was not significantly different between the three treatments.     

Breeding with resistant rams leads to rapid control of classical scrapie in affected sheep flocks

ABSTRACT: Susceptibility to scrapie, a transmissible spongiform encephalopathy in sheep, is modulated by the genetic make-up of the sheep. Scrapie control policies, based on selecting animals of resistant genotype for breeding, have recently been adopted by the Netherlands and other European countries. Here we assess the effectiveness of a breeding programme based on selecting rams of resistant genotype to obtain outbreak control in classical scrapie-affected sheep flocks under field conditions. In six commercially-run flocks following this breeding strategy, we used genotyping to monitor the genotype distribution, and tonsil biopsies and post-mortem analyses to monitor the occurrence of scrapie infection. The farmers were not informed about the monitoring results until the end of the study period of six years. We used a mathematical model of scrapie transmission to analyze the monitoring data and found that where the breeding scheme was consistently applied, outbreak control was obtained after at most four years. Our results also show that classical scrapie control can be obtained before the frequency of non-resistant animals is reduced to zero in the flock. This suggests that control at the national scale can be obtained without a loss of genetic polymorphisms from any of the sheep breeds.     

GC-based analysis of plant stanyl fatty acid esters in enriched foods

Approaches for the capillary gas chromatographic (GC) based analysis of intact plant stanyl esters in enriched foods were developed. Reference compounds were synthesized by enzyme-catalyzed transesterifications. Their identities were confirmed by means of mass spectrometry. Using a medium polar trifluoropropylmethyl polysiloxane stationary phase, long-chain plant stanyl esters could be separated according to their stanol moieties and their fatty acid chains. Thermal degradation during GC analysis was compensated by determining response factors; calibrations were performed for ten individual plant stanyl esters. For the analysis of low-fat products (skimmed milk drinking yogurts), the GC separation was combined with a "fast extraction" under acidic conditions. For fat-based foods (margarines), online coupled LC-GC offered an elegant and efficient way to avoid time-consuming sample preparation steps. The robust and rapid methods allow conclusions on both, the stanol profiles and the fatty acid moieties, and thus provide a basis for the authentication of this type of functional food ingredients.     

Spatial distribution and sample size to estimate Euschistus heros population density in wild plants during off-season

Journal of Plant Diseases and Protection - The neotropical brown stink bug Euschistus heros (F.) is an important pest for many crops in several countries and its ability to survive on associated...     

Method for Precision Nitrogen Management in Spring Wheat: II. Implementation

By accounting for spatial variation in soil N levels, variable-rate fertilizer application may improve crop yield and quality, and N use efficiency within fields. The main purpose of this study was to demonstrate how site-specific wheat yield and protein data, and a geographic information system may be used in developing precision N-recommendations for spring wheat. The three steps in the procedure include: (1) estimate the amount of N-removed in wheat in the year in which the crop is harvested, (2) estimate the N-deficit, defined as the amount of additional N needed for raising protein concentration in a future crop to a specified target level, and (3) estimate the total N-recommendation by summing the mapped values of the N-removed and the N-deficit. A map for variable-rate application of fertilizer is derived by specifying cutoff values to divide the range in the total N-recommendation into classes representing N management zones.A field experiment was conducted within an annually cropped wheat field (101 ha) in northern Montana to determine whether the proposed method could improve grain yields and protein levels. The N-removal and N-deficit were estimated from site-specific wheat yield and protein data that were acquired during harvest of 1996. In 1997, which was a dry year, an experiment was conducted in the same field that consisted of a randomized complete block design arranged as pairs of strip plots. Variable- or uniform-rate N treatments were randomly assigned to each pair of strips. Both treatments received nearly the same amount of fertilizer, however, N in the variable treatment was varied to match patterns in grain yield and protein levels that previously existed in 1996. Yields were not significantly different between management systems, but proteins were significantly enhanced by spatially variable N application. In addition, variability in protein levels was reduced within the whole field. Field areas deficient in N fertility could be identified without having to sample for soil profile N.