Feed or food responsible for the presence of low-level thiouracil in urine of livestock and humans?

In recent years, questions have been raised on the possible semi-endogenous status of the alleged xenobiotic thyreostatic drug thiouracil; thiouracil has been detected in the urine of various animals (livestock and domesticated) at concentrations between 1 and 10 μg L(-1) and also in human urine. Although several studies suggest Brassicaceae-derived feed as potential origin, no traces of thiouracil have been detected in feed so far. Therefore, the aim of this study was to elucidate the origin of thiouracil in the urine of livestock and humans. To this purpose various Brassicaceae feed and food sources (e.g., rapeseed, rapeseed coarse meal, cabbage, cauliflower, broccoli) were investigated for the presence of thiouracil. In addition, the impact of the Brassicaceae-related β-thioglucosidase enzyme was evaluated. This myrosinase enzyme appeared to be crucial, because without its catalyzed hydrolysis no thiouracil could be detected in the various Brassicaceae-derived samples. Therefore, a sample pretreatment with incorporated enzymatic hydrolysis was developed after ensuring the quality performance of the extracted myrosinase mixture with a single-point glucose assay. Upon enzymatic hydrolysis and LC-MS(2) analysis, thiouracil was successfully detected in samples of traditional rapeseed, rapeseed-'00' variety coarse meal (values of erucic acid <2% and glucosinolates <25 μmol g(-1)), and rapeseed cake at 1.5, 1.6, and 0.4 μg kg(-1), respectively. As for the food samples, broccoli and cauliflower displayed thiouracil concentrations of 6.0 and <1.0 μg kg(-1), respectively. To the best of the authors' knowledge this study is the first to report the presence of naturally occurring thiouracil in feed and food samples. Future research should investigate the pathway of thiouracil formation and identify its possible precursors.     

Identification of pathogenic Leptospira strains in tissues of a premature foal by use of polymerase chain reaction analysis.

Studies were carried out to determine the cause of death in a prematurely born Thoroughbred foal that died 24 hours after birth. Necropsy revealed gross lesions suggestive of septicemia. A commercial Leptospira polymerase chain reaction (PCR) assay designed to specifically amplify the hemolysis-associated protein 1 (hap1) gene present only in pathogenic Leptospira strains detected the presence of Leptospira DNA in various tissues of the foal. Histologic examination of lung, liver, kidney, and myocardium revealed numerous spirochetes in Warthin-Starry-stained tissue sections. Results of PCR analysis and histologic examination suggested a leptospiral infection in the newborn foal. At the moment of death, the infection coexisted with a streptococcal-associated aspiration bronchopneumonia and postpartum septicemia. These findings indicate that the PCR assay based on the amplification of the hap1 gene represents a useful tool for specific detection of pathogenic leptospira in field samples taken from horses.     

Soil carbon dioxide efflux in pure and mixed stands of oak and beech

Total Soil Respiration (TSR) was measured in pure and mixed stands of oak and beech and was partitioned into two contributions using the forest floor removal technique: Mineral Soil Respiration (MSR) and Forest Floor Respiration (FFR). In addition, laboratory incubations of the forest floor and the Ah horizon were performed to evaluate the heterotrophic respiration and the DOC production of these horizons. The relationships between soil temperature and the various soil respiration contributions in the three stands were compared using Q ₁₀ functions. In situ, significant differences (α = 0,05) between stands were observed for the R ₁₀ parameter (respiration rate at 10 °C) of the TSR, MSR and FFR contributions, while only the temperature sensitivity (Q ₁₀) of TSR was significantly affected by stand composition. The effect of soil water content was only significant on MSR and followed different patterns according to stand composition. Under controlled conditions, the R ₁₀ of the forest floor and of the Ah horizon varied with stand composition and the Q ₁₀ of the forest floor decreased in the order: oak (2.27) > mixture (2.01) > beech (1.71).     

Prevalence of feline leukemia virus infection and serum antibodies against feline immunodeficiency virus in unowned free-roaming cats

OBJECTIVE: To determine prevalence of FeLV infection and serum antibodies against feline immunodeficiency virus (FIV) in unowned free-roaming cats. DESIGN: Cross-sectional serologic survey. ANIMALS: 733 unowned free-roaming cats in Raleigh, NC, and 1,143 unowned free-roaming cats in Gainesville, Fla. RESULTS: In Raleigh, overall prevalence of FeLV infection was 5.3%, and overall seroprevalence for FIV was 2.3%. In Gainesville, overall prevalence of FeLV infection was 3.7%, and overall seroprevalence for FIV was 4.3%. Overall, FeLV prevalence was 4.3%, and seroprevalence for FIV was 3.5%. Prevalence of FeLV infection was not significantly different between males (4.9%) and females (3.8%), although seroprevalence for FIV was significantly higher in male cats (6.3%) than in female cats (1.5%). CONCLUSIONS AND CLINICAL RELEVANCE: Prevalence of FeLV infection and seroprevalence for FIV in unowned free-roaming cats in Raleigh and Gainesville are similar to prevalence rates reported for owned cats in the United States. Male cats are at increased risk for exposure to FIV, compared with female cats.     

Effect of nitrogen fertilization on nitrogen dynamics in oilseed rape using 15N‐labeling field experiment

In order to optimize nitrogen (N) fertilization and to reduce the environmental impact of oilseed rape without decreasing yield, a clearer understanding of N dynamics inside the plant is crucial. The present investigation therefore aimed to study the effects of different N‐application rates on the dynamics of N uptake, partitioning, and remobilization. The experiment was conducted on winter oilseed rape (Brassica napus L. cv. Capitol) under three levels of N input (0, 100, and 200 kg N ha^–1) from stem elongation to maturity using ¹⁵N‐labeling technique to distinguish between N uptake and N retranslocation in the plant. Nitrogen fertilization affected the time‐course of N uptake and also the allocation of N taken up from flowering to maturity. Most pod N came from N remobilization, and leaves accounted for the largest source of remobilized N regardless the N‐application rate. However, the contribution of leaves to the remobilized N pool increased with the N dose whereas the one of taproot decreased. Stems were the main sink for remobilized N from stem elongation to flowering. Leaves remained longer on N200 than on N0 and N100 plants, and N concentration in fallen leaves increased with the N treatment and in N100 plants along an axial gradient from the basal to the upper leaves. Overall, these results show that the timing of N supply is more crucial than the N amount to attain a high N efficiency.     

Serological studies of Actinobacillus (Haemophilus) pleuropneumoniae strains of serotype 6 and their antigenic relationship with other serotypes

Serological tests such as agglutination, coagglutination, precipitation and indirect haemagglutination were used to study the antigenic relationship of reference and field strains of Actinobacillus (Haemophilus) pleuropneumoniae of serotype 6 with reference strains of other serotypes. Both cell-associated particulate and cell-free soluble antigens prepared from unheated and heat-treated bacterial suspensions of reference and field strains of serotype 6 were used in the studies. Species-specific, common antigenic determinants associated mainly with heat-treated particulate antigens of serotype 6 were cross-reactive in tube agglutination tests with almost all the serotypes. The species-specific antigens were of a minor nature because the cross-reactivities were abolished in both 2-mercaptoethanol agglutination and coagglutination tests. Cell-free saline extracts of both unheated and heat-treated suspensions of serotype 6 strains possessed epitopes specific for serotypes 3, 5 and 8 in addition to their own specific determinants. The epitopes were dominant because the reactions of strains of serotype 6 with antisera against serotypes 3, 5 and 8 persisted in almost all the serological tests used. Serotype 6 strains were antigenically closer to serotype 8 than to serotypes 3 or 5. A combination of serological tests such as coagglutination followed by 2-mercaptoethanol tube agglutination and, or, immunodiffusion tests differentiated serotype 6 strains from those of other cross-reacting serotypes.     

In vitro evaluation of screws and suture anchors in metaphyseal bone of the canine tibia

OBJECTIVE: To compare ease of insertion, load to failure, and mode of failure of cortical and cancellous screws, BoneBiter, IMEX, and TwinFix suture anchors in canine metaphyseal tibial bone. STUDY DESIGN: Experimental biomechanical study. ANIMALS: Canine cadaveric tibias. METHODS: One investigator inserted all anchors and subjectively evaluated ease of placement. Anchor systems were loaded to failure along axis of insertion with audio-video recording to determine failure mode. RESULTS: BoneBiter was the most difficult anchor to insert successfully. Mean+/-SD loads to failure were cancellous screw (711+/-193 N), IMEX 4.7 mm 18 g wire (661+/-163 N), IMEX 4.0 mm 18 g wire (661+/-165 N), cortical screw (635+/-184 N), BoneBiter #5 Kevlar suture (393+/- 109 N), and TwinFix 5.0 mm #2 polyester (267+/-73 N). No significant differences were noted among the cortical screw, cancellous screw, IMEX 4.7 and 4.0 mm, all of which were significantly (P<.001) greater than BoneBiter and TwinFix . Failure modes were pullout of bone, suture-wire breakage, eyelet breakage, or no failure to 1000 N: screws (18,0,0,2), IMEX (18,1,1,0), BoneBiter (2,8,0,0), and TwinFix (0,10,0,0). CONCLUSIONS: Fixation devices were user friendly, with the exception of BoneBiter. Mode of failure is dependent on suture material and anchor design. Cortical and cancellous screws, and IMEX anchors with 18 g wire have significantly greater load to failure compared with BoneBiter and TwinFix suture anchors. CLINICAL RELEVANCE: Based on load to failure, ease of use, design characteristics, and cost, IMEX anchors may have advantages over other comparable soft tissue fixation devices.     

Regulation and function of AtNRAMP4 metal transporter protein

The NRAMP gene family encodes integral membrane proteins mediating the transport of a broad range of transition metals in bacteria, fungi, plants, and animals. We studied the regulation of AtNRAMP4 in Arabidopsis. In a previous study, we showed that AtNRAMP3 and AtNRAMP4 transport manganese (Mn), iron (Fe), and cadmium (Cd). In this study, we show that, in contrast to AtNRAMP3, AtNRAMP4 complements the growth phenotype of the zrt1zrt2 Zn uptake deficient yeast mutant. In a previous study, we have shown that, under Fe starvation, AtNRAMP4 mRNA levels are up-regulated in Arabidopsis. To analyze the regulation of AtNRAMP4 at the protein level, we generated specific antibodies against AtNRAMP4 protein. The antiserum was able to recognize a tagged version of AtNRAMP4 expressed in yeast. The antibody did not reveal any change in AtNRAMP4 protein level upon Fe starvation in Arabidopsis thaliana ecotype Columbia plants. In AtNRAMP4 overexpressing plants, high levels of AtNRAMP4 protein could be detected. AtNRAMP4 overexpressing plants display cadmium hypersensitivity in a medium containing 50 μm FeEDTA as Fe source. However, despite the constitutive accumulation of AtNRAMP4 protein, AtNRAMP4 over-expressing plants did not display Cd hypersensitivity under high Fe supply (100 μm FeHBED). AtNRAMP4 over-expressing lines displayed the same sensitivity to Zn as controls under all conditions tested. Our results suggest a translational level for the regulation of AtNRAMP4. Over-expression of AtNRAMP4 in Arabidopsis thaliana confers a slight hypersensitivity to Cd but not to Zn.