Detergents modify proteinase K resistance of PrP Sc in different transmissible spongiform encephalopathies (TSEs)

Prion diseases are diagnosed by the detection of their proteinase K-resistant prion protein fragment (PrP(Sc)). Various biochemical protocols use different detergents for the tissue preparation. We found that the resistance of PrP(Sc) against proteinase K may vary strongly with the detergent used. In our study, we investigated the influence of the most commonly used detergents on eight different TSE agents derived from different species and distinct prion disease forms. For a high throughput we used a membrane adsorption assay to detect small amounts of prion aggregates, as well as Western blotting. Tissue lysates were prepared using DOC, SLS, SDS or Triton X-100 in different concentrations and these were digested with various amounts of proteinase K. Detergents are able to enhance or diminish the detectability of PrP(Sc) after proteinase K digestion. Depending on the kind of detergent, its concentration - but also on the host species that developed the TSE and the disease form or prion type - the detectability of PrP(Sc) can be very different. The results obtained here may be helpful during the development or improvement of a PrP(Sc) detection method and they point towards a detergent effect that can be additionally used for decontamination purposes. A plausible explanation for the detergent effects described in this article could be an interaction with the lipids associated with PrP(Sc) that may stabilize the aggregates.     

Comparison of viraemia- and clinical-based estimates of within- and between-pen transmission of classical swine fever virus from three transmission experiments

Analyses of recent classical swine fever (CSF) epidemics in the European Union have shown that silent circulation of CSF virus (CSFV) occurs before the first outbreak is detected and this may lead to a large epidemic. However, severity of CSF disease signs may be linked with efficacy of disease transmission, the most severely affected animals having a higher infectivity than the less affected ones. The purpose of this study was to combine disease transmission quantification methods with CSF clinical signs quantification tools to investigate whether clinical signs, considered as infectivity markers, may allow us to calculate reliable estimates for disease transmission parameters. Data from three transmission experiments were used, varying according to the viral strain (Eystrup or Paderborn) and to the contact structure between experimentally inoculated and contact animals (direct or indirect contact). Within- and between-pen basic reproduction ratios (R0) were compared using viraemia data or clinical data. Between-pen R0 estimates were close and not significantly >1, with either strain or computation mode (using viraemia or clinical data). Conversely, within-pen R0s (Paderborn strain) computed using clinical data appeared higher than the estimates obtained using viraemia data. A models comparison (Bayes information criterion) showed a better fit of the clinical-based models, for both strains. This suggests that, in affected herds, the most severely affected animals could play a prominent role in CSFV transmission.     

An easy and rapid method using microscopy to determine herbicide effects in Poaceae weed species

A new, easy, rapid and relatively inexpensive method using microscopy has been developed for the detection of herbicide effects in leaves of grass weed species displaying no visual signs of damage. The method has potential to be used as a tool to indicate future death of grass species due to herbicide exposure by observing phytochemical effects, i.e. early-warning effects. In the present study, Apera spica-venti (L.) Beauv., Bromus hordeaceus L., Alopecurus myosuroides Huds., Lolium perenne L. and Poa annua L. were exposed to lethal rates of four herbicides with different mode of action. The herbicides investigated were the thiocarbamate: prosulfocarb, the sulfonylurea: iodosulfuron, the aryloxyphenoxypropionate: fenoxaprop-P-ethyl and the organophosphate glyphosate. Autofluorescence of leaves was studied under a microscope using ultraviolet and blue light. The fluorescence of leaves treated to enhance flavonoids was also examined. To confirm the results, microspectrofluorometry was performed. Effects indicating future death of the grasses were observed in visually healthy leaves following treatment with prosulfocarb, glyphosate and iodosulfuron. No changes were detected following treatment with fenoxaprop-P-ethyl. After exposure to glyphosate or iodosulfuron, changes in the content of flavonoids and other compounds with a conjugation system and rigid structure and a decrease in the content of chlorophyll were detected in the leaves. Prosulfocarb treatment resulted in changes in the content of flavonoids and other compounds with a conjugation system and rigid structure and an increase in the content of chlorophyll in the leaves. The results obtained from microspectrofluorometry indicated that exposure to prosulfocarb caused a reduction in the flavonoids quercetin, naringenin and/or naringin.     

Environmental,historical, and contextual determinants of vegetation cover: a landscape perspective

We formulated and tested models of relationships among determinants of vegetation cover in two agroforested landscapes of eastern North America (Haut Saint-Laurent, Quebec, Canada) that differed by the spatial arrangement of their geomorphic features and intensity of agricultural activities. Our landscape model compared the woody plots of each landscape in terms of the relative influence of environmental attributes, land use history (1958 – 1997), and spatial context (i.e., proximity of similar or contrasting land cover). Our vegetation model evaluated the relative contribution of the same sets of variables to the distributions of herbs, trees, and shrubs. Relationships were assessed using partial Mantel tests and path analyses. Significant environmental and contextual differences were found between the vegetation plots of the two landscapes, but disturbance history was similar. Our vegetation model confirms the dominant effect of historical factors on vegetation patterns. Whereas land-use history overrides environmental and contextual control for trees, herbaceous and shrub species are more sensitive to environmental conditions. Context is determinant only for understory species in older, less-disturbed plots. Results are discussed in relevance to vegetation dynamics in a landscape perspective that integrates interactions between environmental and human influences.     

Assessing the agronomic potential of linseed genotypes by multivariate analyses and association mapping of agronomic traits

High prices of fish oil make linseed attractive for aquaculture and animal feed. To ensure a constant supply of linseed, the development of stable cultivars is of strategic importance. In this study, 35 linseed genotypes were evaluated in five Chilean environments (E) from 2009 to 2012. The additive main effect and multiplicative interaction analysis (AMMI), genotype (G) plus genotype by environment (GE) interaction (GGE) biplot analysis and three stability parameters were tested with the aim of identifying adapted genotypes for the development of linseed cultivars. An association mapping (AM) analysis was also conducted for four agronomic traits and the stability of the associated markers was evaluated using the QQE (QTL main effect and QTL by environment interaction) approach. Combined analysis of variance for yield, seeds per boll (SPB), plant height (PH) and days to flowering (DTF) were significant for G, E and GE (P < 0.001). The combined stability analysis identified some Canadian, Argentinean and Chilean accessions to be the best adapted and highest yielding genotypes. Coancestry analysis indicated that crossing Canadian and Chilean genotypes could maximize transgressive segregation for yield. Significant associations for DTF, PH and SPB explained up to 59 % of the phenotypic variation for these traits. The QQE and AM analyses were consistent in identifying marker LGM27B as the most stable and significant across all environments with the largest effect in reducing DTF. The combined application of the stability, AM and QQE analyses could accelerate the development of marketable linseed cultivars adapted to Southern Chile.     

Comparison of sandy soils suppressive or conducive to ectoparasitic nematode damage on sugarcane

ABSTRACT Two South African sandy soils, one suppressive and the other conducive to ectoparasitic nematode damage on monoculture sugarcane, were compared. Analysis of field transects indicated that the suppressive soil displayed a comparatively higher population of the weak ectoparasite Helicotylenchus dihystera, whose predominance among ectoparasitic nematodes is known to limit yield loss caused by more virulent phytonematodes. Soil type was identical at both sites (entisols), but the suppressive soil had a higher organic matter content and a lower pH, which correlated with H. dihystera population data. In contrast, microclimatic differences between the two field sites were unlikely to be responsible for the suppressive or conducive status of the soils, as shown in a greenhouse experiment. The two soils exhibited a bacterial community of the same size but with different genetic structures, as indicated by automated ribosomal intergenic spacer analysis (RISA). The number of culturable fluorescent pseudomonads was higher for the conducive soil, probably because extensive root damage caused by ectoparasitic nematodes favored proliferation of these bacteria. This study shows that apparently small differences in soil composition between fields located in the same climatic area and managed similarly can translate into contrasted nematode communities, ectoparasitic nematode damage levels, and sugarcane yields.     

The Brucella genome at the beginning of the post-genomic era

The year 2002 began with the publication of the first complete genome sequence for a Brucella species, that of the two replicons of B. melitensis 16M. Hopefully in 2002, the complete genome of B. suis 1330, and, perhaps, a B. abortus strain will be published. This is the culmination of over 30 years investigation of the composition, structure, organisation and evolution of the Brucella genome. Brucella research must now adapt to the new challenges of the post-genomic era.