PCR as a diagnostic and quantitative technique in veterinary parasitology

Over the past 15 years, there has been a dramatic evolution in molecular approaches to study parasites and parasitic diseases. Many of these advancements have been brought about through the development of new applications of the polymerase chain reaction (PCR). Enhancements in sensitivity that can be achieved using PCR now permit scientists to investigate changes at the level of a single cell, far below what is often needed for parasite-derived applications. PCR has had a substantial impact on advances made in the areas of parasite systematics and epidemiology, immunology and host–parasite interactions, recombinant DNA vaccine development and most re- cently, the analysis of whole genomes either through directly sequencing the DNA, the analysis of expressed sequence tags (ESTs) or through the rapidly growing field of functional genomics. This paper, however, focuses on the application of PCR methodology to parasite detection and differen- tiation, and the diagnosis of disease. Specific attention is given to advances provided by multiplex PCR, fluorescence-based “real-time” PCR, and the utilization of PCR as a quantitative technique. Published by Elsevier Science B.V

VALIDITY OF 95% t-CONFIDENCE INTERVALS UNDER SOME TRANSECT SAMPLING STRATEGIES

Soil pH data were used to assess the capture rates of 95 % t-confidence intervals based on five different transect sampling strategies. Two different sampling methods were considered, deterministic and two-stage simple random sampling . The data used were pH readings at 15 and 30 centimeter depths from two local agricultural fields in the Manhattan, Kansas area. The data provided three distinct populations with three different distributions - skewed left, symmetric, and bimodal. The total number of transects randomly sampled was 2, 5, and 10. The total number of points sampled along each transect was 2, 7 and 14. The 95% t-confidence intervals were simulated 5000 times using five different transect sampling strategies, and the capture rates of the population mean were recorded. Box plots of the capture rates for the five transect sampling strategies were constructed and compared. In most cases the deterministic sampling method had capture rates that underestimated the 95 % confidence level, whereas the two-stage sampling strategies produced capture rates which were conservative or closer to the 95 % confidence level. The variances of the capture rates for the two-stage sampling strategies were relatively small in comparison to the deterministic sampling strategies. In conclusion, the two-stage simple random sampling method along with the Satterthwaite degrees of freedom are recommended when using transect sampling

Effects of supplementation of limit-fed growing diets with either soybean meal or nonenzymatically browned soybean meal on steer performance

Seventy two individually fed Angus × Hereford steers (642 lb) were used to evaluate the effects of supplementing limit-fed, growing diets with either soybean meal (SBM) or nonenzymatically browned soybean meal (NSBM). Eight steers were allotted to a control diet composed of 39.1% high-moisture corn, 42% cottonseed hulls, 10.4% ground corn, 5% cane molasses 2.25% urea, and 1.5% vitamins and minerals (dry basis). The remaining steers were allotted to diets that derived 100, 80, 60, or 40% of their supplemental protein from SBM or 60, 45, 30, or 15% of their supplemental protein from NSBN. The balance of supplemental protein came from urea. All diets were formulated to contain 13.0% crude protein (dry basis). Steers were fed once daily for 80 days at 2.25% of BW. Average daily gain and efficiency did not differ (P>.05) between sources (ADG=1.932 + .103 × (% CP from SBM) + .097 × (% CP from NSBM); gain:feed=.140 + .0058 × (% CP from SBM) + .0051 × (% CP from NSBM)). The lack of response to NBSBM supplementation above that for SBM suggests that either degradable intake protein was limiting in the basal diet or a large proportion of the amino acids in the NSBM were unavailable due to overprocessing

COMPARING MEASURING METHODS USING THE SENSITIVITY RATIO: AN APPLICATION TO RESISTANCE SCREENING IN SOYBEANS

When there are several methods of measuring a physical or chemical property, it is necessary to determine which method is best. If both methods are measured on the same scale, the most precise method will be preferred. However, often the methods have different scales. The sensitivity ratio allows for explicit comparison of methods with different scales. We use the sensitivity ratio to compare soybean resistance screening methods to evaluate the resistance of soybean varieties to Sclerotinia sclerotiorum. When compared to the root mean square error or the coefficient of variation, the sensitivity ratio can order methods differently both when the methods are measured on different scales and on the same scale. Our results cast doubt upon using standard precision statistics such as the root mean square error or the coefficient of variation to compare measuring methods and we suggest that the sensitivity ratio should be used instead

Dakota Gold®-brand dried distiller’s grains with solubles: effects on finishing performance and carcass characteristics

A 153-day trial was conducted using 345 heifers to determine optimal level of Dakota Gold dried distiller’s grains with solubles (DDGS) in finishing diets based on steam-flaked corn. Diets contained six levels of DDGS: 0%, 15%, 30%, 45%, 60%, and 75%. DDGS affected average daily gain, final weight and hot carcass weight, all of which increased with 15% DDGS and then decreased as additional DDGS was added. Growth performance of heifers fed 30% DDGS was similar to those fed no DDGS. In general, heifers were overfinished, with 61% being Yield Grade 3 or greater and 83% grading Choice or Prime. Backfat tended to decrease with addition of DDGS, and kidney, pelvic, and heart fat and marbling scores tended to be greatest for intermediate levels of DDGS. Percentage of carcasses grading Choice or Prime tended to be lower for heifers fed 60 or 75% DDGS

Measurement of molecular mixing at a conjugated polymer interface by specular and off-specular neutron scattering

Measurements have been performed on thermally equilibrated conjugated-polymer/insulating-polymer bilayers, using specular and off-specular neutron reflectivity. While specular reflectivity is only sensitive to the structure normal to the sample, off-specular measurements can probe the structure of the buried polymer/polymer interface in the plane of the sample. Systematic analysis of the scattering from a set of samples with varying insulating-polymer-thickness, using the distorted-wave Born approximation (DWBA), has allowed a robust determination of the intrinsic width at the buried polymer/polymer interface. The quantification of this width (12 Å ± 4 Å) allows us to examine aspects of the conjugated polymer conformation at the interface, by appealing to self-consistent field theory (SCFT) predictions for equilibrium polymer/polymer interfaces in the cases of flexible and semi-flexible chains. This analysis enables us to infer that mixing at this particular interface cannot be described in terms of polymer chain segments that adopt conformations similar to a random walk. Instead, a more plausible explanation is that the conjugated polymer chain segments become significantly oriented in the plane of the interface. It is important to point out that we are only able to reach this conclusion following the extensive analysis of reflectivity data, followed by comparison with SCFT predictions. It is not simply the case that conjugated polymers would be expected to adopt this kind of oriented conformation at the interface, because of their relatively high chain stiffness. It is the combination of a high stiffness and a relatively narrow intrinsic interfacial width that results in a deviation from flexible chain behaviour

A survey of phytoestrogenic activity in Kansas flint hills pastures

The botanical composition and basal cover of three Kansas Flint Hills pastures located in Butler and Chase counties was surveyed to estimate the incidence of plant species that contain appreciable levels of estrogenic activity. Many-flowered scurfpea and Ladino clover were the only plant species classified as high in estrogenic activity. Although significant estrogenic activity existed in specific species, the willingness of livestock to consume those species is unclear

Acoustic Energy and Momentum in a Moving Medium

By exploiting the mathematical analogy between the propagation of sound in a non-homogeneous potential flow and the propagation of a scalar field in a background gravitational field, various wave ``energy'' and wave ``momentum'' conservation laws are established in a systematic manner. In particular the acoustic energy conservation law due to Blokhintsev appears as the result of the conservation of a mixed co- and contravariant energy-momentum tensor, while the exchange of relative energy between the wave and the mean flow mediated by the radiation stress tensor, first noted by Longuet-Higgins and Stewart in the context of ocean waves, appears as the covariant conservation of the doubly contravariant form of the same energy-momentum tensor.Comment: 25 Pages, Late