Enhanced cutaneous hypersensitivity reactions are associated with ovine high and low cortisol responsiveness to acute endotoxin challenge

Inbred rodent studies have demonstrated that cutaneous hypersensitivity reactions are exacerbated in stress-susceptible, and attenuated in stress-resistant strains of mice. This physiological response was, in part, mediated by activation of the hypothalamic-pituitary-adrenal axis during the acute restraint stress. A study was conducted to examine whether or not cutaneous hypersensitivity reactions are also associated with variable cortisol responsiveness to inflammatory stress in an outbred ovine population. High (H), medium (M), and low (L) cortisol responsive sheep were identified from a population of 110 females based on their estimated breeding values for cortisol concentration measured 4 h post-systemic challenge with Escherichia coli endotoxin (400 ng kg(-1)). Cutaneous hypersensitivity reactions to phytohaemagglutinin (PHA), 1-chloro-2, 4-dinitrobenzene (DNCB), and Candida albicans cellular antigen (CAA) were measured in these variable cortisol-responding sheep, in addition to serum interleukin (IL)-6, interferon (IFN)-gamma, and ovalbumin (OVA)-specific IgG concentrations. When compared to the M cortisol responders, both H and L cortisol responders had significantly greater cutaneous swelling during the elicitation phase in response to DNCB (P < 0.05) and CAA (P < 0.05); a similar but not significant trend was observed during the PHA challenge. The primary, but not the secondary, IgG response to OVA was significantly lower in the H and L cortisol responders when compared to the M cortisol responders. Differences in serum IL-6 or IFN-gamma concentration were not observed across variable cortisol-responsive groups. Together, these results demonstrate that cutaneous hypersensitivity reactions are enhanced in outbred H and L cortisol-responding sheep, independent of systemic modulation by IL-6 and IFN-gamma.     

Effect of the US3 protein of bovine herpesvirus 5 on the actin cytoskeleton and apoptosis

The US3 protein is a unique protein kinase only present in the Alphaherpesvirinae subfamily of the herpesviruses. Studies performed with several alphaherpesviruses demonstrated that the US3 protein is involved in cytoskeleton modifications during viral infection and displays anti-apoptotic activity. However, the US3 protein of BoHV-5 has not been studied up to now. As reported for other alphaherpesviruses, our results showed that BoHV-5 US3 confers resistance against apoptosis and induces cytoskeletal reorganization leading to cell rounding, actin stress fiber breakdown and cell projections that interconnect cells. The expression of a kinase-dead version of BoHV-5 US3 showed that the anti-apoptotic activity and the induction of cell projections are kinase-dependent whereas kinase activity is not absolutely required for actin stress fiber breakdown. Besides, the kinase-dead version of US3, but not the wild type protein, was found excluded from the nucleus. These results constitute the first report on the BoHV-5 US3 functions, and highlight that there are functional differences and similarities among US3 proteins of different alphaherpesviruses.     

Antibody-induced internalization of viral glycoproteins in pseudorabies virus-infected monocytes and role of the cytoskeleton: a confocal study

Addition of pseudorabies virus (PrV)-specific polyclonal immunoglobulins to PrV-infected monocytes induces internalization of plasma membrane anchored viral glycoproteins. This process may interfere with antibody-dependent cell lysis and resembles the well-studied physiological endocytosis process. A confocal study was designed to investigate whether the major cellular components, involved in physiological endocytosis (clathrin, actin, dynein and microtubules), play a role in this virological internalization process. In order to visualize the interaction of endosomes, which contain the internalized viral glycoproteins, with clathrin, actin, dynein and microtubules, a double labeling of viral glycoproteins and different cellular proteins was performed. Porcine monocytes were inoculated with the PrV-strain 89V87 at a multiplicity of infection of 50 for 13h. After the addition of FITC-labeled porcine polyclonal PrV-specific antibodies, cells were fixed with para-formaldehyde at different time points and afterwards permeabilized. The different cellular components were visualized with monoclonal antibodies and a Texas Red-conjugate, with the exception of actin, which was stained with phalloidin-Texas Red. The cells were analyzed by confocal microscopy. A clear co-localization was observed between the viral glycoproteins and clathrin and dynein during the internalization process. The microtubules were in close contact with the internalized vesicles. For actin no co-localization could be observed. It can be stated that clathrin, dynein and microtubules, important components during physiological endocytosis, are also of importance during the antibody-induced internalization of viral glycoproteins.     

The Influence of a Particle Separation Technology on the Generation of Airborne Particles from Different Roughages and Bedding Materials Used for Horses

Four different types of bedding materials (wheat straw, wood shavings, hemp shives, flax shives) and two roughages (hay, haylage) were treated using an air-driven particle separation technology. The airborne particle and mold generation of both treated and untreated samples were then analyzed under standardized laboratory conditions. In addition, samples of all the treated materials were stored for 8 weeks either in a pressed or incoherent form and then analyzed again for their ability to generate airborne particles. The airborne particle concentrations were detected online with the gravimetrically measuring analyzer tapered element oscillating microbalance (TEOM) 1400a that was equipped successively with different inlets to measure the particle fractions PM₂₀, PM₁₀, PM_2.5, and PM_1.0 (PM = Particulate matter). The particle separation resulted in a reduction in the airborne particle (PM₂₀) generation in all materials: hay 49.16 to 22.79 mg/m³ (53.6%), haylage 28.57 to 25.04 mg/m³ (12.3%), wood shavings 141.68 to 15.04 mg/m³ (89.4%), wheat straw 143.08 to 22.97 mg/m³ (83.9%), flax 135.11 to 53.31 mg/m³ (60.5%), and hemp 63.67 to 17.64 mg/m³ (72.3%). The 8-week storage of the treated materials as compressed materials led to a renewed significant increase in the airborne particle (PM₁₀) concentration in the haylage (+29.9%), wheat straw (+104.0%), wood shavings (+40.4%), and hemp shives (+30.7%). Storage of the incoherent materials caused a significant increase in these particles only in the wheat straw (+44.2%). The separation treatment reduced the mold production by 92.4% in the wood shavings, 88.0% in the wheat straw, and 85.8% in the hay.     

Genetic diversity of livestock-associated MRSA isolates obtained from piglets from farrowing until slaughter age on four farrow-to-finish farms

During a previous longitudinal study, performed on four farrow-to-finish farms (A to D), samples were taken from twelve sows, their offspring, and the environment on various occasions over six months to study the MRSA presence. During the present study, a selection of the obtained MRSA isolates were typed by multiple-locus variable-number tandem-repeat analysis (MLVA), Pulsed Field Gel Electrophoresis (PFGE), spa typing, and SCCmec typing to study the genetic diversity of LA-MRSA isolates and to determine possible MRSA sources for pig(let)s. PFGE, spa typing, and SCCmec typing revealed the presence of one or few dominant genotype(s) per farm. In contrast, 212 MLVA types were detected on the four farms, forming one cluster on farm A, three on farm B, four on farm C and two on farm D. The genotype, found on farm A was unique for this farm. Farms B, C and D shared one cluster. In general, MLVA types from these clusters were isolated from piglets, sows, and the environment on various sampling events. Piglets carried MLVA types both related and unrelated to their mother sows’ MLVA types at farrowing and onwards. In conclusion, molecular typing revealed that within a farm one or a few dominant strain(s) are widespread. Potential MRSA sources for piglets were mother sows, the environment and other piglets.

Electronic supplementary material

The online version of this article (doi:10.1186/s13567-014-0089-4) contains supplementary material, which is available to authorized users.     

Antimicrobial use in Belgian broiler production

The use of antimicrobials in production animals has become a worldwide concern in the face of rising resistance levels in commensal, pathogenic and zoonotic bacteria. In the years 2007 and 2008 antimicrobial consumption records were collected during two non consecutive production cycles in 32 randomly selected Belgian broiler farms. Antimicrobials were used in 48 of the 64 monitored production cycles, 7 farms did not use any antimicrobials in both production cycles, 2 farms only administered antimicrobials in one of the two production cycles, the other 23 farms applied antimicrobial treatment in both production cycles. For the quantification of antimicrobial drug use, the treatment incidences (TI) based on the defined daily doses (the dose as it should be applied: DDD) and used daily doses (the actual dose applied: UDD) were calculated. A mean antimicrobial treatment incidence per 1000 animals of 131.8 (standard deviation 126.8) animals treated daily with one DDD and 121.4 (SD 106.7) animals treated daily with one UDD was found. The most frequently used compounds were amoxicillin, tylosin and trimethoprim-sulphonamide with a mean TI(UDD) of 37.9, 34.8, and 21.7, respectively. The ratio of the UDD/DDD gives an estimate on correctness of dosing. Tylosin was underdosed in most of the administrations whereas amoxicillin and trimethoprim-sulphonamide were slightly overdosed in the average flock.     

Milk quality assurance for paratuberculosis: simulation of within-herd infection dynamics and economics

A bulk milk quality assurance programme for Mycobacterium avium subsp. paratuberculosis (Map) in dairy herds was simulated with a stochastic simulation model (JohneSSim). The aim of this study was to evaluate the epidemiological and economic effects of preventive management measures and various test schemes in a simulated population of closed Dutch dairy herds over a 20-year period. Herds were certified as ;low-Map bulk milk' if, with a certain probability, the concentration of Map in bulk milk did not exceed a maximum acceptable concentration of 10(3) Map organisms per litre (based on pasteurisation studies). The programme started with an initial assessment; test-negative herds entered a surveillance procedure and test-positive herds a control procedure. The simulations showed that herd examinations by ELISA for the initial assessment, surveillance and control procedures effectively ensure the quality of ;low-Map bulk milk': > 75% of simulated herds were certified and > 96% of certified herds produced bulk milk with < 10(3) Map/L if the initial herd-level prevalence was 30%. Preventive management measures only had a minor effect on bulk milk quality of certified herds. Culling based on biennial faecal culture was more effective than culling based on annual ELISA. Average total discounted costs for 20-year participation in a programme consisting of initial assessment by ELISA, surveillance by biennial ELISA and control by biennial faecal culture were 16 Euro x 10(3) per herd. In conclusion, this study shows that a bulk milk quality assurance programme for closed Dutch dairy herds is feasible and provides information on the cost-effectiveness of different programmes. The concepts of this study equally apply to other countries because mechanisms of paratuberculosis infection, disease, and testing are comparable in other dairy cattle populations.