We analyse the results of our experimental laboratory approximation of motorway networks with slime mould Physarum polycephalum. Motorway networks of 14 geographical areas are considered: Australia, Africa, Belgium, Brazil, Canada, China, Germany, Iberia, Italy, Malaysia, Mexico, the Netherlands, UK and USA. For each geographical entity, we represented major urban areas by oat flakes and inoculated the slime mould in a capital. After slime mould spanned all urban areas with a network of its protoplasmic tubes, we extracted a generalised Physarum graph from the network and compared the graphs with an abstract motorway graph using most common measures. The measures employed are the number of independent cycles, cohesion, shortest paths lengths, diameter, the Harary index and the Randić index. We obtained a series of intriguing results, and found that the slime mould approximates best of all the motorway graphs of Belgium, Canada and China, and that for all entities studied the best match between Physarum and motorway graphs is detected by the Randić index (molecular branching index).

Belgium is amongst few artificial countries, established on purpose, when Dutch and French speaking parts were joined in a single unit. This makes Belgium a particularly interesting testbed for studying bio-inspired techniques for simulation and analysis of vehicular transport networks. We imitate growth and formation of a transport network between major urban areas in Belgium using the acellular slime mould Physarum polycephalum. We represent the urban areas with the sources of nutrients. The slime mould spans the sources of nutrients with a network of protoplasmic tubes. The protoplasmic tubes represent the motorways. In an experimental laboratory analysis we compare the motorway network approximated by P. polycephalum and the man-made motorway network of Belgium. We evaluate the efficiency of the slime mould network and the motorway network using proximity graphs.

Bluetongue has emerged recently in Belgium. A bluetongue virus strain was isolated and characterized as serotype 8. Two new real-time reverse transcription-quantitative PCRs (RT-qPCRs) that amplified 2 different segments of bluetongue virus detected this exotic strain. These 2 RT-qPCRs detected infection earlier than a competitive ELISA for antibody detection.

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.

The need for fast and very early detection of foot-and-mouth disease virus (FMDV) infection has yielded different types of diagnostic tools over the past decades: whereas very sensitive techniques such as virus isolation (VI) and more recently also real-time RT-PCR can provide evidence for the presence of low virus quantities, VI requires additional confirmation of the nature of the virus strain and both techniques (currently) lack the ability for direct serotyping. The latter usually depends on ELISA, which is a far less sensitive method and may require virus culturing. This paper elaborates on experimental efforts towards the development of an 'immuno-rolling circle amplification (RCA)' assay in 96-well plates, the aim being to increase the sensitivity of immunological FMDV detection and serotyping by means of RCA. The study attempts to explain the encountered hurdles and the complexity of the different setups tested. Conclusively, immuno-RCA in 96-well plates as a reliable diagnostic assay for FMDV seems very difficult to achieve.

Soil erosion on agricultural land and its detrimental environmental and economical effects has aroused increased interest among both the research and policy-making communities. The call for erosion control measures adapted to local farming practices is high, especially in Europe where farmers are reluctant to adopt soil conservation techniques. This study investigates a new technique for controlling concentrated flow erosion rates in the loess belt of central Belgium: i.e. double drilling of cereals in zones of concentrated flow. Cross-sectional areas of erosion channels as well as crop yield parameters in single- and double-drilled zones were compared. The technique is based on the combined effect of the increased density of plant shoots and roots for reducing soil loss. Results indicate that double drilling can reduce soil loss through concentrated flow by 25% on average and by up to 40% under optimal conditions. No net change in wheat grain yield was observed, and farmers who participated in the experiments were satisfied with the results and the easy application of the technique. Globally, benefits were larger than costs. However, the effectiveness of the technique in reducing soil loss by concentrated flow erosion seems to be topographically restricted. For positions in the landscape with a contributing drainage area larger than ca. 0.75ha, the effectiveness of double drilling can be doubted. Double drilling should therefore be regarded as one possibility amongst others to reduce concentrated flow erosion rates in farmers’ fields.

Summary Farmers in Europe want to control soil erosion in ways that are easily incorporated in their normal practices. We have investigated the possibility of reducing soil erosion by concentrated flow (i.e. rill and gully erosion) through increasing the root density of cereal crops. In situ root density measurements on cereal fields were combined with laboratory flume experiments on samples, taken in single‐ and double‐drilled fields, of which the above‐ground biomass was clipped. During the laboratory experiments no significant effect of root densities on critical shear stress or channel erodibility was observed because of interactions with other changing parameters (e.g. ageing effects). Therefore, the expected relative detachment rates as a function of plant root density were calculated using an empirical equation. During the first 75 days of the crop growth season relative soil detachment rates for single‐drilled field parcels can be reduced up to 50% compared with a rootless field, whereas relative soil detachment rates in double‐drilled field parcels can be reduced up to 60% in this period. Thereafter, plant roots in double‐drilled field parcels reduce relative soil detachment rates on average by 9% compared with single‐drilled field parcels (up to an absolute maximum of 90% compared with rootless soils). During the growing season, not only root density increases but also the vegetation cover changes, which enhances soil protection from erosion. Therefore, cereal roots will help to conserve the soil when seed is drilled at double rates, especially during the early growth stages and in fields with medium risk of concentrated flow.

The twin-arginine translocation (Tat) system exports folded proteins across bacterial cytoplasmic membranes. Recently, genes encoding TatA, TatB and TatC homologues were identified in Streptomyces lividans and the functionality of the Tat pathway was demonstrated. Here, we have examined the localization and structural organization of the Tat components in S. lividans. Interestingly, besides being membrane-associated proteins, S. lividans TatA and TatB were also detected in the cytoplasm. TatC could only be detected in isolated membrane fractions. Whereas all TatC was found to be stably inserted in the membrane, part of membrane-associated TatA and TatB could be extracted following high salt, sodium carbonate or urea treatment suggesting a more loose association with the membrane. Finally, we have analyzed Tat complexes that could be purified from an S. lividans TatABC overproducing strain. From the cytoplasmic membrane, two types of high molecular mass Tat complexes could be isolated having a similar composition as those isolated from Escherichia coli. In the cytoplasm, TatA and TatB were detected as monomer or as homo-oligomeric complexes.

Several molecular techniques require high quality RNA, completely free of DNA. Standard methods to isolate total RNA from Streptomyces spp. are based on the application of a ‘Modified Kirby Mix’ [Practical Streptomyces Genetics. The John Innes Foundation, Norwich, pp. 613]. Here we present an alternative procedure using Triton® X-100 and EDTA for the isolation of total RNA from Streptomyces.

The entire double-stranded DNA genome of the Streptomyces venezuelae bacteriophage VWB was sequenced and analyzed. Its size is 49,220 bp with an overall molar G + C content of 71.2 mol%. Sixty-one potential open reading frames were identified and annotated using several complementary bioinformatics tools. Clusters of functionally related putative genes were defined, supporting a refined version of the modular theory of phage evolution.

Streptomycetes are attractive microorganisms because of their high secretion capacity and for the production of secondary metabolites. We report the improvement of a PCR-based method for screening of solid media-grown Streptomyces transformants. By resuspending mycelium into 2.5-10% dimethyl sulfoxide (DMSO), extensive manipulation prior to PCR could be avoided, thus significantly reducing screening time. Results proved to be both reliable and specific.