Identification of ALS inhibitor‐resistant Amaranthus biotypes using polymerase chain reaction amplification of specific alleles

Summary Polymerase chain reaction (PCR) amplification of specific alleles (PASA) was adapted as a molecular marker‐based method for the rapid detection of point mutations in Amaranthus retroflexus and Amaranthus rudis leading to ALS inhibitor resistance. Two pairs of primers were designed for the specific amplification of alleles of the ALS gene of susceptible and resistant biotypes. The allele‐specific primer matched the desired allele, but mismatched the different allele at its 3′ end. Differentiation was carried out by comparison of the amplified DNA fragments in gel electrophoresis after PASA‐PCR. In A. rudis, differentiation was possible with one PCR and genomic DNA as probe. A ‘nested’ PCR was necessary for the differentiation of sensitive and resistant A. retroflexus. PASA is useful for the identification of resistant weed biotypes and also as a monitoring tool to map resistance occurrence and distribution. Advantages include the fast and clear separation of those plants with and without mutations at an early stage of development, its easy and consistent performance and quick results compared with existing resistance detection tests. These advantages, when combined with management strategies, enable further activities to reduce herbicide resistance.     

Ranking cows’ methane emissions under commercial conditions with sniffers versus respiration chambers

This study assessed the ranking of dairy cows using individual-level correlations for methane (CH₄) emission on-farm using sniffers and in respiration chambers. In total 20 lactating dairy cows, ten Holstein and ten Jerseys were recorded using sniffers installed in milking robots for three weeks of lactation and subsequently in respiration chambers (RC) where they were each recorded on three occasions within the RC. Bivariate linear mixed models were used to determine the individual-level correlations (r_I) between sniffer and RC phenotypes as proxies for genetic correlations. Despite differences in feeding and management, the predicted CH₄ production from sniffers correlated highly with RC CH₄ production r_I?=?0.77?±?0.18 and CH₄ breath concentration correlated nearly as well with RC CH₄ production r_I?=?0.75?±?0.20. These correlations between sniffers on-farm and RC demonstrate the potential of sniffers measurements as large-scale indicator traits for CH₄ emissions in dairy cattle.     

Pharmacokinetics of xylazine after 2-, 4-, and 6-hr durations of continuous rate infusions in horses

Intravenous (i.v.) bolus administration of xylazine (XYL) (0.5 mg/kg) immediately followed by a continuous rate infusion (CRI) of 1 mg kg⁻¹ hr⁻¹ for 2, 4, and 6 hr produced immediate sedation, which lasted throughout the duration of the CRI. Heart rate decreased and blood pressure increased significantly (p > .05) in all horses during the first 15 min of infusion, both returned to and then remained at baseline during the duration of the infusion. Compartmental models were used to investigate the pharmacokinetics of XYL administration. Plasma concentration–time curves following bolus and CRI were best described by a one-compartment model. No differences were found between pharmacokinetic estimates of the CRIs for the fractional elimination rate constant (K_e), half-life (t_1/2e), volume of distribution (V_d), and clearance (Cl). Median and range were 0.42 (0.15–0.97)/hr, 1.68 (0.87–4.52) hr, 5.85 (2.10–19.34) L/kg, and 28.7 (19.6–39.5) ml min⁻¹ kg⁻¹, respectively. Significant differences were seen for area under the curve ( ) (p < .0002) and maximum concentration (C_max) (p < .04). This indicates that with increasing duration of infusion, XYL may not accumulate in a clinically relevant way and hence no adjustments are required in a longer XYL CRI to maintain a constant level of sedation and a rapid recovery.     

Expert opinion based modelling of the risk of human infection with H5N1 through the consumption of poultry meat in Germany

According to current scientific opinion the risk of human infection with H5N1 via preparation and consumption of poultry meat is negligible.This opinion has not yet been challenged by a formal risk assessment, due to the lack of empirical data. We have developed a scenario pathway model as a conceptual framework for a formal assessment of the H5N1 risk to humans through consumption of poultry meat and parameterise the model using information derived from expert opinions. The aim of this study was to investigate whether the notion of an overall negligible risk via the oral infection route is consistent with ad hoc data and expert opinions on the relevant parameters of the model.The model is mainly based on expert opinion. A stochastic Monte-Carlo simulation was conducted which took into consideration (amongst others) the exposure and infection of chicken (broiler and layer), turkeys, ducks and geese, the probabilities of detection prior to slaughter, virus survival and contamination during slaughter, as well as during the cutting and preparation of meat in commercial plants and in private households, respectively. The empirical consumption pattern for poultry meat in Germany was taken into account in the simulation.The results show that the risk for the individual consumer is practically zero whereas up to 23 cases per year in Germany might occur if the upper (more pessimistic) ranges of the expert opinions apply.The finding of a low but non-negligible risk to the population is discussed in relation to the epidemiological information available from recent outbreaks in South East Asia.     

High-resolution ultrasonography of xenografted domestic cat ovarian cortex

Transplantation of ovarian tissue has high potential for female gamete conservation. However, optimal timing of oocyte recovery for in vitro maturation and fertilization is still critical. Therefore the aim of the present study was to use high-resolution transcutaneous ultrasonography to monitor follicular development within xenografted ovarian tissue. Ovarian cortex fragments (n=44) from domestic cats were transplanted into athymic nude rats (n=12). Graft development in the animals was assessed weekly by high frequency ultrasound (10-22 MHz) under two different FSH regimes. Blood collection for serum estradiol determination and vaginal smears were performed simultaneously. The xenografts were removed at different time points according to the ultrasound findings. The survival rate of the transplants 4 weeks after surgery was 54.5% and antral follicular growth was observed within 10 grafts from 5 different hosts (8.6 +/- 6.43 follicles per graft). Early follicle antrums could be detected from 0.4 mm onwards. The growth rate of the antral cavity was calculated from weekly measurements (0.56 +/- 0.44 mm per week). Although vaginal cells and estradiol levels followed a cyclic pattern, no correlation was found between follicular diameter, estradiol and keratinized vaginal cells. We recovered 5, 1 and 4 cumulus oocyte complexes from three different individuals during weeks 19, 21, and 23 respectively. Extrusion of a polar body (1 oocyte) and germinal vesicle break down (7 oocytes) indicated progression of maturation after in vitro culture. We conclude that ultrasonography und provided a reliable method to examine xenograft survival and follicular development within the grafts. Furthermore, this technique is suitable for assessment of the efficiency of hormonal treatment and narrowing of the optimal time frame for oocyte retrieval. To our knowledge, this is the first report of the in vivo development of early antral follicles in mammalian species.     

Application of urease inhibitor improves protein composition and bread‐baking quality of urea fertilized winter wheat

Background: Nitrogen losses is an economic problem for wheat production and a high risk to the environment. Therefore, improved N fertilizer management is a key to increasing the N efficiency and minimizing N losses. To increase N efficiency, enhanced fertilizers such as urea combined with urease inhibitor can be used. Aims: The aim of present study was to evaluate the effects of different N forms on grain storage protein subunits in winter wheat and to examine whether the observed changes correlate with parameters of baking quality. Methods: The investigation was performed over two consecutive years at two locations in Germany. Protein subunits were analyzed by SDS‐PAGE. Results: Protein concentrations were similarly increased after fertilization with ammonium nitrate and urea + urease inhibitor. Analysis of the individual storage protein fractions indicated that both fertilizers specifically enhanced ω‐gliadins and HMW glutenins, but the effect was more pronounced in the ammonium nitrate treatment. Application of urea + urease inhibitor had greater influence on the protein composition and resulted in higher specific baking volume as well as the best fresh keeping ability, in comparison with urea treatment. Conclusion: Considering that the urea + urease inhibitor treatment resulted in almost comparable improvements of NUE and baking quality, with the additional benefit of reduced N losses in combination with easy handling, urea + urease inhibitor can be recommended as a viable alternative to both urea alone and ammonium nitrate treatments. This opens up an opportunity for the reduction of N loss in wheat production when use of urea is preferred.     

Stabilization mechanisms of organic matter in four temperate soils: Development and application of a conceptual model

Based on recent findings in the literature, we developed a process‐oriented conceptual model that integrates all three process groups of organic matter (OM) stabilization in soils namely (1) selective preservation of recalcitrant compounds, (2) spatial inaccessibility to decomposer organisms, and (3) interactions of OM with minerals and metal ions. The model concept relates the diverse stabilization mechanisms to active, intermediate, and passive pools. The formation of the passive pool is regarded as hierarchical structured co‐action of various processes that are active under specific pedogenetic conditions. To evaluate the model, we used data of pool sizes and turnover times of soil OM fractions from horizons of two acid forest and two agricultural soils. Selective preservation of recalcitrant compounds is relevant in the active pool and particularly in soil horizons with high C contents. Biogenic aggregation preserves OM in the intermediate pool and is limited to topsoil horizons. Spatial inaccessibility due to the occlusion of OM in clay microstructures and due to the formation of hydrophobic surfaces stabilizes OM in the passive pool. If present, charcoal contributes to the passive pool mainly in topsoil horizons. The importance of organo‐mineral interactions for OM stabilization in the passive pool is well‐known and increases with soil depth. Hydrophobicity is particularly relevant in acid soils and in soils with considerable inputs of charcoal. We conclude that the stabilization potentials of soils are site‐ and horizon‐specific. Furthermore, management affects key stabilization mechanisms. Tillage increases the importance of organo‐mineral interactions for OM stabilization, and in Ap horizons with high microbial activity and C turnover, organo‐mineral interactions can contribute to OM stabilization in the intermediate pool. The application of our model showed that we need a better understanding of processes causing spatial inaccessibility of OM to decomposers in the passive pool.