Evidence of excretion of Schmallenberg virus in bull semen

Schmallenberg virus (SBV) is a novel orthobunyavirus, discovered in Germany in late 2011. It mainly infects cattle, sheep and goats and could lead to congenital infection, causing abortion and fetal abnormalities. SBV is transmitted by biting midges from the Culicoides genus and there is no evidence that natural infection occurs directly between ruminants. Here, we could detect SBV RNA in infected bull semen using qRT-PCR (three bulls out of seven tested positive; 29 positive semen batches out of 136). We also found that highly positive semen batches from SBV infected bulls can provoke an acute infection in IFNAR^-/- mice, suggesting the potential presence of infectious virus in the semen of SBV infected bulls.     

Spatial distribution of lineages of oak powdery mildew fungi in France, using quick molecular detection methods

? Powdery mildew is a major fungal disease of oaks in Europe. Recent studies using internal transcribed spacer (ITS) sequences suggested the presence of four different lineages (putative species). The objective of the study was to investigate the spatial distribution of these lineages/species and, in particular, to test the hypothesis of a spatial differentiation, at various scales: regional (France), altitudinal (a Pyrenean transect) and local (within a forest plot).

? Detection methods for the four ITS types were developed: (1) single strand conformation polymorphism analysis (SSCP); (2) PCR amplifications, for which specific primers were designed. SSCP proved to be efficient for the detection of Erysiphe alphitoides and E. quercicola types. In contrast, the rarer ITS types corresponding to E. hypophylla and Phyllactinia guttata (sensu lato) were only detected by specific amplification.

? The study confirmed the strong predominance of the ITS sequence associated with E. alphitoides at all spatial scales (with a frequency higher than 80%). Isolates presumably belonging to E. quercicola (i.e. with same ITS type), a recently described species not yet recorded in Europe, were also found in all French regions at a significant frequency (15% at national level).

? No pattern of spatial differentiation between the putative species could be demonstrated: E. alphitoides was often found in association with different ITS types in the same region, the same tree, and even in the same lesion.

     

Changes in quantitative patterns of dead wood in maritime pine plantations over time

Although the volume of dead wood is commonly acknowledged as an indicator of biodiversity in sustainable forest management schemes, only few data are available for plantation forests. To evaluate the volume and qualify the diversity of woody debris along a chronosequence of maritime pine plantations, we sampled downed woody debris, snags and stumps in 143 stands of different ages. To test the hypothesis that the pattern of dead wood accumulation mainly results from silvicultural operations, we developed a predictive model. It combined an empirical growth model evaluating the amount of dead wood produced by successive thinnings with a decay function that estimated the loss of dead wood with time.     

Dynamic MRI and thermal simulation to interpret deformation and water transfer in meat during heating

Understanding and controlling structural and physical changes in meat during cooking is of prime importance. Nuclear magnetic resonance imaging (MRI) is a noninvasive, nondestructive tool that can be used to characterize certain properties and structures both locally and dynamically. Here we show the possibilities offered by MRI for the in situ dynamic imaging of the connective network during the cooking of meat to monitor deformations between 20 and 75 °C. A novel device was used to heat the sample in an MR imager. An MRI sequence was developed to contrast the connective tissue and the muscle fibers during heating. The temperature distribution in the sample was numerically simulated to link structural modifications and water transfer to temperature values. The contraction of myofibrillar and collagen networks was observed at 42 °C, and water began to migrate toward the interfascicular space at 40 °C. These observations are consistent with literature results obtained using destructive and/or nonlocalized methods. This new approach allows the simultaneous monitoring of local deformation and water transfer, changes in muscle structure and thermal history.     

Detection of Diplodia sapinea in Corsican pine seeds

Diplodia sapinea is one of the major pathogens of pines worldwide. Despite the putative critical importance of seed infection in the epidemiology of the disease, this aspect of the biology of the fungus is poorly known. Here, biological and molecular methods were developed for the detection of the fungus and applied to assess D. sapinea infection in Corsican pine seeds. A buffered medium containing tannic acid and malt extract as a nutrient base was the most efficient and selective for D. sapinea recovery. A molecular method based on DNA extraction with a commercial kit and specific amplification, including an internal amplification control, was developed. A high percentage of infection (57% positive isolations) was observed in seeds obtained from fallen cones in a Corsican pine stand with no apparent symptoms of D. sapinea. Seeds collected from trees in a seed orchard showing severe symptoms of dieback caused by D. sapinea had comparatively lower infection (38%). Moreover, very low infection levels (1–5%) were observed after the standard treatment used for seed extraction, which included heating at 40°C. Diplodia sapinea was not recovered from seedlings grown from infected seed lots submitted to water stress. Overall, results suggest that the risk of disease transmission by commercial seeds is probably low, but could be further reduced by thermotherapy.     

Colostral antibody induced interference of inactivated bluetongue serotype-8 vaccines in calves

Since its introduction into northern Europe in 2006, bluetongue has become a major threat to animal health. While the efficacy of commercial vaccines has been clearly demonstrated in livestock, little is known regarding the effect of maternal immunity on vaccinal efficacy. Here, we have investigated the duration and amplitude of colostral antibody-induced immunity in calves born to dams vaccinated against bluetongue virus serotype 8 (BTV-8) and the extent of colostral antibody-induced interference of vaccination in these calves. Twenty-two calf-cow pairs were included in this survey. The median age at which calves became seronegative for BTV was 84 and 112 days as assayed by seroneutralisation test (SNT) and VP7 BTV competitive ELISA (cELISA), respectively. At the mean age of 118 days, 13/22 calves were immunized with inactivated BTV-8 vaccine. In most calves vaccination elicited a weak immune response, with seroconversion in only 3/13 calves. The amplitude of the humoral response to vaccination was inversely proportional to the maternal antibody level prior to vaccination. Thus, the lack of response was attributed to the persistence of virus-specific colostral antibodies that interfered with the induction of the immune response. These data suggest that the recommended age for vaccination of calves born to vaccinated dams needs to be adjusted in order to optimize vaccinal efficacy.     

Influence of culture medium pH on internalization, growth and phenotypic plasticity of Neospora caninum

Neospora caninum, a strictly intracellular protozoan, is a major leading cause of parasite-induced abortion in cattle. A widely held view of N. caninum infection is that both cellular proliferation and stage interconversion (tachyzoite-bradyzoite transformation) are triggered, perhaps even modulated by, changes in cultural conditions. This study tested the hypothesis that exposure of N. caninum tachyzoites to different pH culture media affects the parasite's entry, proliferation and cyst formation in cultured cells. The endocytic pathway for N. caninum entry into the K562 cell line was found to be mediated by low pH of culture medium. Internalization of N. caninum by host cells was significantly increased in acidic and alkaline culture medium compared to cells maintained in neutral medium as revealed by transmission electron microscopy. Parasite proliferation within Vero cells was assessed by plaque formation assay and was found to be highest when pH level was optimum, paralleled by a decrease in the number of cysts. In contrast, parasite encystation increased when the pH level was alkaline or acidic, as evaluated by indirect immunofluorescence and immunocytochemical analyses. Acidic pH regardless of state of host cell infection suppressed the rate of host cell division. These findings suggest that culture medium pH has a determinable effect on the host cell-N. caninum interaction and support the hypothesis that pH of culture medium influence the entry, growth, and phenotypic plasticity of N. caninum in mammalian cells.