Association between serum cobalamin and methylmalonic acid concentrations in dogs

The aim of this study was to evaluate the association between serum methylmalonic acid (MMA), a proposed marker of cellular cobalamin deficiency, and serum cobalamin concentrations in dogs. Serum samples from 555 dogs were grouped according to their serum cobalamin concentrations (<150 ng/L to 1000 ng/L). Additionally, serum samples were collected from 43 healthy dogs to calculate a reference interval for canine serum MMA. MMA was measured using a GC/MS method. Groups were compared using a Kruskal-Wallis test with Dunn's post test. Proportions of dogs above the upper limit of the reference interval were calculated and a χ2-test for trend was performed to evaluate the association between serum cobalamin and MMA concentrations. The reference interval for serum MMA was calculated to be 414.7-1192.5 nmol/L. Dogs with serum cobalamin concentrations <251 ng/L had significantly higher MMA concentrations (P<0.05) and the χ2-test for trend showed a trend for increasing serum MMA concentrations with decreasing serum cobalamin concentrations (P<0.0001). Additionally, a number of dogs with normal serum cobalamin concentrations had increased serum MMA concentrations, suggesting that some of these dogs may have cobalamin deficiency on a cellular level. Further studies are warranted to determine if these dogs should receive cobalamin supplementation.     

Genotyping of Bacillus anthracis Isolated from Croatia and Bosnia and Herzegovina

Anthrax is a serious disease caused by Bacillus anthracis. Humans can become infected by handling products from infected animals, by breathing spores and rarely by eating undercooked meat from infected animals. The genome of B. anthracis is highly monomorphic and thus shows very low DNA sequence variation. We analysed the molecular characteristics of 12 B. anthracis isolates from outbreaks in Croatia and Bosnia and Herzegovina, which have occurred during the past 10 years along with two vaccine strains. Genotyping system based on variable‐number tandem repeat analysis at six loci revealed that six isolates belong to genotype from the A1.a cluster whilst six isolates relate to the B2 cluster, compared to 89 previously described genotypes. The distribution of two evolutionarily distant clusters suggests an introduction of B. anthracis to this area in at least two separate events.     

Nitrogen retention and plant uptake on a highly weathered central Amazonian Ferralsol amended with compost and charcoal

Leaching losses of N are a major limitation of crop production on permeable soils and under heavy rainfalls as in the humid tropics. We established a field trial in the central Amazon (near Manaus, Brazil) in order to study the influence of charcoal and compost on the retention of N. Fifteen months after organic‐matter admixing (0–0.1 m soil depth), we added ¹⁵N‐labeled (NH₄)₂SO₄ (27.5 kg N ha^–1 at 10 atom% excess). The tracer was measured in top soil (0–0.1 m) and plant samples taken at two successive sorghum (Sorghum bicolor L. Moench) harvests. The N recovery in biomass was significantly higher when the soil contained compost (14.7% of applied N) in comparison to only mineral‐fertilized plots (5.7%) due to significantly higher crop production during the first growth period. After the second harvest, the retention in soil was significantly higher in the charcoal‐amended plots (15.6%) in comparison to only mineral‐fertilized plots (9.7%) due to higher retention in soil. The total N recovery in soil, crop residues, and grains was significantly (p < 0.05) higher on compost (16.5%), charcoal (18.1%), and charcoal‐plus‐compost treatments (17.4%) in comparison to only mineral‐fertilized plots (10.9%). Organic amendments increased the retention of applied fertilizer N. One process in this retention was found to be the recycling of N taken up by the crop. The relevance of immobilization, reduced N leaching, and gaseous losses as well as other potential processes for increasing N retention should be unraveled in future studies.     

Ferulic acid from aleurone determines the antioxidant potency of wheat grain (Triticum aestivum L.)

Grain is an important source of phytochemicals, which have potent antioxidant capacity. They have been implicated in the beneficial health effect of whole grains in reducing cardiovascular disease and type 2 diabetes. The aim of the present study was to identify the most important antioxidant fractions of wheat grain. It was found that the aleurone content of these fractions was highly correlated with the antioxidant capacity of the fractions (r = 0.96, p < 0.0001). Ferulic acid appeared to be the major contributor to the antioxidant capacity in fractions with higher antioxidant capacity. The contribution of protein was rather limited. It was concluded that the antioxidant potency of wheat grain fractions is predominantly determined by aleurone content, which can be attributed to the presence of relatively large amounts of phenolic compounds, primarily ferulic acid.     

Charcoal and smoke extract stimulate the soil microbial community in a highly weathered xanthic Ferralsol

The influence of charcoal and smoke condensates (pyroligneous acid, PA) on microbial activity in a highly weathered Amazonian upland soil was assessed via measurements of basal respiration (BR), substrate-induced respiration (SIR), and exponential population increase after substrate addition. PA extracts are commonly used for fertilizer or as pest control in Brazil, where phosphorus (P) availability and nitrogen (N) leaching are among the most severe limitations for agriculture. Microbes play an important role in nutrient cycling and solubilizing of phosphate. BR, microbial biomass, population growth and the microbe's efficiency (expressed by the metabolic quotient) increased linearly and significantly with increasing charcoal concentrations (50, 100 and 150 g kg⁻¹ soil). Application of PA caused a sharp increase in all parameters. We suppose that the condensates from smoke contain easily degradable substances and only small amounts of inhibitory agents, which could be utilized by the microbes for their metabolism.     

Bioactive constituent production in St. John's Wort in vitro hairy roots. Regenerated plant lines

A wild strain of Agrobacterium rhizogenes was used to regenerate twelve in vitro plant lines from different hairy roots of H. perforatum (St. John's Wort). The production of the main bioactive constituents was observed even though their yields varied in the different plant lines. Two lines were selected for the hyperoside production (4.9-4.6 mg/gdw) while nine were characterized by significant yields of chlorogenic acid (ranged from 0.47 to 1.09 mg/gdw). Furthermore, one out of twelve lines showed a 10-fold higher hypericin content (0.25 mg/gdw) than that reported for the in vitro shoots in the literature. Morphological and phytochemical features were determined in order to select H. perforatum genotypes enriched in valuable bioactive compounds.     

Evaluation of antibody-ELISA and real-time RT-PCR for the diagnosis and profiling of bluetongue virus serotype 8 during the epidemic in Belgium in 2006

In 2006 bluetongue (BT) emerged for the first time in North-Western Europe. Reliable diagnostic tools are essential in controlling BT but data on the diagnostic sensitivity (Se) and specificity (Sp) are often missing. This paper aims to describe and analyse the results obtained with the diagnostics used in Belgium during the 2006 BT crisis. The diagnosis was based on a combination of antibody detection (competitive ELISA, cELISA) and viral RNA detection by real-time RT-PCR (RT-qPCR). The performance of the cELISA as a diagnostic tool was assessed on field results obtained during the epidemic and previous surveillance campaigns. As the infectious status of the animals is unknown during an epidemic, a Bayesian analysis was performed. Both assays were found to be equally specific (RT-qPCR: 98.5%; cELISA: 98.2%) while the diagnostic sensitivity of the RT-qPCR (99.5%) was superior to that of the cELISA (87.8%). The assumption of RT-qPCR as standard of comparison during the bluetongue virus (BTV) epidemic proved valid based on the results of the Bayesian analysis. A ROC analysis of the cELISA, using RT-qPCR as standard of comparison, showed that the cut-off point with the highest accuracy occurred at a percentage negativity of 66, which is markedly higher than the cut-off proposed by the manufacturer. The analysis of the results was further extended to serological and molecular profiling and the possible use of profiling as a rapid epidemiological marker of the BTV in-field situation was assessed. A comparison of the serological profiles obtained before, during and at the end of the Belgian epidemic clearly showed the existence of an intermediate zone which appears soon after BTV (re)enters the population. The appearance or disappearance of this intermediate zone is correlated with virus circulation and provides valuable information, which would be entirely overlooked if only positive and negative results were considered.