First detection of kobuvirus in farm animals in Brazil and the Netherlands.

Animal kobuviruses have been described in pigs, cattle, sheep and bats in countries in Asia and Europe. The virus can be detected in fecal and serum samples of infected animals with or without diarrhea, but most of the clinical as well as epidemiological features of kobuvirus infection are still unknown. This study reports the first detection of kobuvirus in farm animals from Brazil and the Netherlands and the molecular analysis of the detected strains. In Brazil, 53% (61/115) of the pigs (suckling, weaned and sows) were shedding porcine kobuvirus in feces, while in the Netherlands 16.7% (3/18) of the tested weaned pigs were infected. Kobuviruses detected in fecal samples of pigs in Brazil showed association (p = 0.0002) with diarrhea. In pig serum, kobuvirus was detected at different ages (3, 21, 36, 60, 75, and 180 days), with an overall rate of 76.7% (23/30). The sequencing of amplicons detected in serum of pigs of different ages suggested reinfection and no persistent infection. Kobuvirus was also detected in sheep and cattle feces from Brazil and the Netherlands, respectively. Phylogenetic analyses of Brazilian and Dutch kobuviruses from pig, cattle and sheep revealed genetic variability, particularly in one strain detected in sheep feces, which was more closely related to human Aichi virus. The molecular and phylogenetic analyses performed with other published kobuvirus strains and the strains presented in this study, showed that, in most of the cases, kobuvirus seems to group according to host species, but not to geographical region of origin. The data presented in this study contribute to the comprehension of kobuvirus epidemiology and also to the molecular identification of kobuvirus strains circulating worldwide

Geostrophic convective turbulence: The effect of boundary layers

Rayleigh--B\'enard (RB) convection, the flow in a fluid layer heated from below and cooled from above, is used to analyze the transition to the geostrophic regime of thermal convection. In the geostrophic regime, which is of direct relevance to most geo- and astrophysical flows, the system is strongly rotated while maintaining a sufficiently large thermal driving to generate turbulence. We directly simulate the Navier--Stokes equations for two values of the thermal forcing, i.e. $Ra=10^{10}$ and $Ra=5\cdot10^{10}$, a constant Prandtl number~$Pr=1$, and vary the Ekman number in the range $Ek=1.3\cdot10^{-7}$ to $Ek=2\cdot10^{-6}$ which satisfies both requirements of super-criticality and strong rotation. We focus on the differences between the application of no-slip vs. stress-free boundary conditions on the horizontal plates. The transition is found at roughly the same parameter values for both boundary conditions, i.e. at~$Ek\approx 9\times 10^{-7}$ for~$Ra=1\times 10^{10}$ and at~$Ek\approx 3\times 10^{-7}$ for~$Ra=5\times 10^{10}$. However, the transition is gradual and it does not exactly coincide in~$Ek$ for different flow indicators. In particular, we report the characteristics of the transitions in the heat transfer scaling laws, the boundary-layer thicknesses, the bulk/boundary-layer distribution of dissipations and the mean temperature gradient in the bulk. The flow phenomenology in the geostrophic regime evolves differently for no-slip and stress-free plates. For stress-free conditions the formation of a large-scale barotropic vortex with associated inverse energy cascade is apparent. For no-slip plates, a turbulent state without large-scale coherent structures is found; the absence of large-scale structure formation is reflected in the energy transfer in the sense that the inverse cascade, present for stress-free boundary conditions, vanishes.Comment: Submitted to JF

Low-threshold Care for Marginalised Hard Drug Users: Marginalisation and Socialisation in the Rotterdam Hard Drug Scene

Since the early 1990s several developments have taken place in the hard drug scene in the Netherlands. Key elements in these developments were harm reduction measures, introduction of crack, open drug scenes, police interventions, drug-related nuisance, low-threshold care facilities and the well-being of drug users. Drug policy and drug scenes have also changed in the past 15 years. The aim of this thesis is to provide insight into the role of low-threshold care facilities in the process of marginalisation and socialisation of drug users. Marginalisation entails that chronic hard drug users drift away from the core institutions of society, e.g. family and friends, the labour market and health care; socialisation is the opposite of marginalisation (Coumans, 2005). The roles that low-threshold care facilities can play are derived from this process. First, low-threshold care facilities can contribute to the discontinuation of marginalisation. Second, low-threshold care facilities can function as re-integration instruments. Both marginalisation and socialisation have their effects on individual drug users and on society in a broader perspective (in terms of public health and public safety). By understanding the role of low-threshold care facilities within a changing environment, these facilities can be utilised more effectively. First, this chapter outlines the concepts of ‘hard drug scenes and nuisance’ and ‘harm reduction and low-threshold care’. Then, Coumans’ theory of marginalisation and socialisation is presented and the six chapters of this thesis are introduced. Finally, the methodology and data used are presented

Visible diode lasers can be used for flow cytometric immunofluorescence and DNA analysis

This report describes a feasibility study concerning the use of a visible diode laser for two important fluorescence applications in a flow cytometer. With a 3 mW 635 nm. diode laser, we performed immunofluorescence measurements using the fluorophore allophycocyanin (APC). We have measured CD8 positive lymphocytes with a two-step labeling procedure and the resulting histograms showed good separation between the negative cells and the dim and the bright fluorescent subpopulations. As a second fluorescence application, we chose DNA analysis with the recently developed DNA/ RNA stains TOTO-3 and TO-PRO-3. In our setup TO-PRO-3 yielded the best results with a CV of 3.4%. Our results indicate that a few milliwatts of 635 nm light from a visible diode laser is sufficient to do single color immunofluorescence measurements with allophycocyanin and DNA analysis with TO-PRO-3. The major advantages of using a diode laser in a flow cytometer are the small size, the low price, the high efficiency, and the long lifetime

Genetic lines differ in Toll-like receptor gene expression in spleens of chicks inoculated with Salmonella enterica serovar Enteritidis

Toll-like receptors (TLR) recognize evolutionarily conserved molecular motifs (pathogen-associated molecular patterns) of infectious microbes and initiate innate immune response upon activation with relevant pathogens. This study investigated the acute effect of Salmonella Enteritidis challenge on TLR mRNA expression in cecum and spleen of birds from 3 distinct genetic lines. Chicks from broiler, Leghorn, and Fayoumi lines were inoculated or mock-inoculated with Salmonella Enteritidis. The mRNA expression levels of TLR2, TLR4, and TLR5 genes were assessed by quantitative reverse transcription-PCR of cecum and spleen tissue harvested at 2 or 18 h postinoculation (PI). There were no significant genetic line effects on TLR mRNA expression in spleen or cecum of mock-infected birds, or in the cecum of infected birds. Genetic line effect was significant (P \u3c 0.05) on TLR mRNA expression in the spleen of Salmonella Enteritidis- infected birds. The Fayoumi line had higher TLR2 and TLR4 expression than Leghorn, higher TLR2 mRNA expression than broiler, and the broiler line had higher TLR5 expression than Leghorn and Fayoumi. In Salmonella Enteritidis-infected birds, the TLR2 expression in both cecum and spleen and TLR4 expression in spleen were significantly higher at 18 h PI than 2 h PI. The results demonstrate a significant genetic line effect on TLR expression in the spleen of Salmonella Enteritidis-infected birds, which may partly explain genetic variability in immune response to Salmonella Enteritidis