Myelin proteolipid protein (PLP) as a marker antigen of central nervous system contaminations for routine food control

Spreading transmissible spongiform encephalopathies (TSE) have been widely attributed to transmission by ingestion of mammalian central nervous system (CNS) tissue. Reliable exclusion of this epidemiological important route of transmission relies on an effective surveillance of food contamination. Here, myelin proteolipid protein (PLP) is identified as a specific and largely heat-resistant marker for detection of food contaminations by CNS tissue. PLP is a component of oligodendritic glial sheaths of neuronal processes that is specifically expressed in the CNS. A highly selective polyclonal antibody was developed directed against an epitope present in the full-length PLP protein, but absent from the developmentally regulated splice variant DM-20. In combination with a hydrophobic extraction of PLP from tissue samples, the antibody reliably detected PLP from spinal cord, cerebellum, and cortex of different mammalian species. Consistent with earlier reports on PLP expression, no cross-reactivity was observed with peripheral nerve or extraneural tissue, except for a very faint signal obtained with heart. When applied to an artificial CNS contamination present in sausages, the antibody reliably detected a low concentration (1%) of the contaminant. Application of heat, as used during conventional sausage manufacturing, led to a predominant alteration of arginine residues in the PLP protein and a partial loss of immunoreactivity. In contrast, a stretch of hydrophilic amino acids(112-122) proved to be heat-resistant, preserving the immunogenicity of this PLP epitope during heating. Taken together, the excellent CNS specificity of PLP immunodetection and the presence of a heat-resistant epitope have permitted the development of a highly sensitive immunoassay for CNS contamination in routine food control.     

Epidemiological enquiries in two Q fever outbreaks in a community of Baden-Württemberg during 2008 and 2009

In 2008 and 2009, two consecutive outbreaks of Q fever in humans were recorded in the district of Freudenstadt, northern Black Forrest, Baden-Württemberg, Germany. In 2008, a total of 41 persons from a single local community fell ill and were found infected with Coxiella burnetii. Although comprehensive diagnostic and epidemiological outbreak investigations were conducted and control measures taken which included vaccination of ruminants at risk in three parts of the affected community, re-occurrence of the disease in 2009 with further 29 confirmed human Q fever cases could not be prevented. While the origin of infection of the first outbreak was probably a flock of 550 sheep moved in the surrounding of the affected villages, the source of infection for the consecutive outbreak in 2009 could not be identified. It seems possible that meadows contaminated with infectious placenta or birth fluids represented the sources of infection.     

Characterization of anthocyanins and pyranoanthocyanins from blood orange [Citrus sinensis (L.) Osbeck] juice

Anthocyanins from blood orange [Citrus sinensis (L.) Osbeck] juices were isolated and purified by means of high-speed countercurrent chromatography and preparative high-performance liquid chromatography. Structures of the pigments were then elucidated by electrospray ionization multiple mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy. The major anthocyanins of the juice were characterized as cyanidin 3-glucoside and cyanidin 3-(6"-malonylglucoside). Furthermore, six minor anthocyanins were detected and identified as cyanidin 3,5-diglucoside, delphinidin 3-glucoside, cyanidin 3-sophoroside, delphinidin 3-(6"-malonylglucoside), peonidin 3-(6"-malonylglucoside), and cyanidin 3-(6"-dioxalylglucoside). The occurrence of the latter compound in blood oranges is reported here for the first time, together with full NMR spectroscopic data. Further investigations revealed the presence of four anthocyanin-derived pigments, which are formed through a direct reaction between anthocyanins and hydroxycinnamic acids during prolonged storage of the juice. These novel pyranoanthocyanins were identified as the 4-vinylphenol, 4-vinylcatechol, 4-vinylguaiacol, and 4-vinylsyringol adducts of cyanidin 3-glucoside through comparison of their mass spectrometric and chromatographic properties with those of synthesized reference compounds.     

In vitro potential antioxidant activity of (1-->3),(1-->6)-beta-D-glucan and protein fractions from Saccharomyces cerevisiae cell walls

(1-->3),(1-->6)-Beta-D-Glucan, a cell wall polysaccharide in many microorganisms, fungi and algae, is a well-known biological response modifier. Recently, it was found that (1-->3)-beta-D-glucan from Saccharomyces cerevisiae also exhibits antioxidative capabilities. In this study the antioxidative activity of the cell wall fractions of brewer's yeast was investigated. Particular emphasis was put on the question to which extent glucan is responsible for the antioxidative activity of the cell walls and how the other cell wall components might contribute. For the experiments yeast cell walls from brewery fermentations were used. Glucan was isolated by a three-step extraction procedure including a combination of hot water and enzymatic treatment. The level of (1-->3),(1-->6)-beta-D-glucan in the cell walls was analyzed enzymatically. The antioxidant activity was determined by electron paramagnetic resonance spectrometry and Trolox equivalent antioxidant capacity assay. The results show that the antioxidative activity of yeast cell wall proteins exceeds that of beta-glucan greatly. Especially aromatic side chains and free thiols from denatured proteins seem to work as antioxidants.     

Manipulation of host hepatocytes by the malaria parasite for delivery into liver sinusoids

The merozoite stage of the malaria parasite that infects erythrocytes and causes the symptoms of the disease is initially formed inside host hepatocytes. However, the mechanism by which hepatic merozoites reach blood vessels (sinusoids) in the liver and escape the host immune system before invading erythrocytes remains unknown. Here, we show that parasites induce the death and the detachment of their host hepatocytes, followed by the budding of parasite-filled vesicles (merosomes) into the sinusoid lumen. Parasites simultaneously inhibit the exposure of phosphatidylserine on the outer leaflet of host plasma membranes, which act as "eat me" signals to phagocytes. Thus, the hepatocyte-derived merosomes appear to ensure both the migration of parasites into the bloodstream and their protection from host immunity.     

Differences in the immunopathogenesis of infectious bursal disease virus (IBDV) following in ovo and post-hatch vaccination of chickens

Not much is known about IBDV-pathogenesis and immune mechanisms following in ovo vaccination. In this study, we compared the immunopathogenesis of an intermediate IBDV-vaccine in post-hatch- and in ovo-inoculated chickens. In ovo-vaccinated birds recovered significantly faster from lesions of the bursa of Fabricius than post-hatch vaccinated (P<0.05). A significant accumulation of intrabursal CD8(+) T cells was observed in post-hatch but not in in ovo-vaccinated chickens (P<0.05). The innate immunity was comparable between in ovo- and post-hatch-vaccinated groups as indicated by comparable intrabursal macrophage accumulation and intrabursal IBDV-clearance. Overall, our observations indicate that IBDV in ovo vaccination may be advantageous over post-hatch. In ovo-vaccinated birds recover faster from bursa lesions and exhibit similar protection against challenge in comparison to post-hatch vaccinated.     

Soil properties of drained and rewetted fen soils

The intensive agricultural use and consequently the drainage of fen soils have caused modifications in structure and nutrient dynamics. Pedogenetic processes result in the formation of typical soil horizons with distinctive soil properties. These are the basis for soil classification. In the present review, results are compiled. Modifications of abiotic and biotic parameters of fen soils due to drainage and rewetting are presented. Recommendations on the further use of fen soils are submitted.     

Soil fauna modifies the recalcitrance-persistence relationship of soil carbon pools

Traditional models of soil organic matter decomposition predict that soil carbon pools with high chemical stability and large physical structure are more resistant against degradation than chemically labile and fine-grained material. We investigated whether soil fauna, by its direct and indirect effects on carbon turnover, would reinforce or counteract this general trend.The effects of four major faunal groups on carbon pools of differing recalcitrance were studied in an extensive microcosm experiment. Ninty-six microcosms were inoculated with nematodes, enchytraeids, collembola, and lumbricids in three densities, including combinations of groups. Bare agricultural soil and soil covered with maize litter were used as substrates. The microcosms were kept under constant conditions at 12 °C and 50% water holding capacity for 60 days. At the end of the experiment, soil particles were separated into size classes (<63 μm, 63-250 μm, >250 μm) and carbon pools were separated into solubility fractions (K₂SO₄-soluble, pyrophosphate-soluble, insoluble), by means of ultrasonic dispersion and subsequent stepwise solubilisation.Both in bare soil and in soil with litter, the carbon pools with the highest chemical stability (insoluble) and the larger particle sizes (>63 μm) were degraded more intensively than all other pools in the presence of lumbricids. The pools of intermediate chemical stability (pyrophosphate-soluble) underwent simultaneous degradation and neoformation brought about by different animal groups. The chemically most labile pool (K₂SO₄-soluble) remained largely unaffected by the fauna. Fixation of carbon in microbial biomass was increased by nematodes in bare soil and by enchytraeids in soil with litter. The results illustrate in detail how, under the influence of soil fauna, soil carbon pools are decomposed in a cascade-like process where carbon is transferred from the stable to the more labile pools, while simultaneously a proportion is fixed in microbial biomass and another part is lost as CO₂. Thereby, the relationship between a substrate's persistence and its chemical stability and physical size is substantially modified. We summarize the mechanisms that most likely are responsible for the different effects of the investigated faunal groups.