Towards middle-up analysis of polyclonal antibodies: subclass-specific N-glycosylation profiling of murine immunoglobulin G (IgG) by means of HPLC-MS.

In recent years, advanced HPLC-MS strategies based on intact protein ("top-down") or protein subunit ("middle-up/middle-down") analysis have been implemented for the characterization of therapeutic monoclonal antibodies. Here, we assess feasibility of middle-up/middle-down analysis for polyclonal IgGs exhibiting extensive sequence variability. Specifically, we addressed IgGs from mouse, representing an important model system in immunological investigations.

Design of Phl p 6 Variants With Altered Fold-Stability Significantly Impacts Antigen Processing, Immunogenicity and Immune Polarization.

Understanding, which factors determine the immunogenicity and immune polarizing properties of proteins, is an important prerequisite for designing better vaccines and immunotherapeutics. While extrinsic immune modulatory factors such as pathogen associated molecular patterns are well-understood, far less is known about the contribution of protein inherent features. Protein fold-stability represents such an intrinsic feature contributing to immunogenicity and immune polarization by influencing the amount of peptide-MHC II complexes (pMHCII).

Mast cells and γδ T cells are largely dispensable for adaptive immune responses after laser-mediated epicutaneous immunization.

Background: The skin resembles an attractive target for vaccination due to its accessibility and abundance of resident immune cells. Cells like γδ T cells and mast cells (MCs) are part of the first line of defence against exogenous threats. Despite being important mediators for eliciting TH2 immune responses after epithelial stress, γδ T cell and MC functions still remain to be completely understood.

Teaching parasitology in a modular veterinary curriculum - The Vienna experience.

In a changing world with rapidly evolving new technologies, even in the "sheltered world" of teachers and lecturers at universities, we are challenged by new developments. As diversification is increasing in many aspects of our professional life, there is also a need to employ new ways of teaching, learning and assessments in veterinary curricula, taking into account the increasing numbers of students and limited teaching resources. The "classical" Austrian veterinary curriculum, with a series of consecutive lectures followed by practical courses in the different disciplines, separated from each other in the curricular time table and by annual examinations, has been a long-standing concept for teaching in the past.

A systematic review of financial and economic assessments of bovine viral diarrhea virus (BVDV) prevention and mitigation activities worldwide.

Infection with bovine viral diarrhea virus (BVDV) results in major economic losses either directly through decreased productive performance in cattle herds or indirectly, such as through expenses for control programs. The aim of this systematic review was to review financial and/or economic assessment studies of prevention and/or mitigation activities of BVDV at national, regional and farm level worldwide. Once all predefined criteria had been met, 35 articles were included for this systematic review.

Evaluation of competence-based teaching in higher education: From theory to practice.

Competence-based teaching in higher education institutions and its evaluation have become a prevalent topic especially in the European Union. However, evaluation instruments are often limited, for example to single student competencies or specific elements of the teaching process. The present paper provides a more comprehensive evaluation concept that contributes to sustainable improvement of competence-based teaching in higher education institutions.

Red deer as maintenance host for bovine tuberculosis, Alpine region.

To estimate the prevalence of bovine tuberculosis in the Alpine region, we studied the epidemiology of Mycobacterium caprae in wildlife during the 2009-2012 hunting seasons. Free-ranging red deer (Cervus elaphus) were a maintenance host in a hot-spot area, mainly located in Austria.

First detection of hepatitis E virus in Austrian pigs by RT-qPCR.

Long known to cause disease outbreaks in man in countries with poor sanitary conditions, an increasing number of autochthonous HEV genotype 3 infections have been reported in industrialised countries. Genotype 3 poses an important potential zoonotic threat, with infected pigs functioning as the main reservoir. This study reports the first detected emergence of HEV in Austrian pigs.

Role of Vpma phase variation in Mycoplasma agalactiae pathogenesis.

Compared with other bacterial pathogens, the molecular mechanisms of mycoplasma pathogenicity are largely unknown. Several studies in the past have shown that pathogenic mycoplasmas are equipped with sophisticated genetic systems that allow them to undergo high-frequency surface antigenic variations. Although never clearly proven, these variable mycoplasma surface components are often implicated in host immune evasion and adaptation.

[Evaluation of four commercial RT-qPCR test kits for detection of Bluetongue virus RNA].

Since 2006, the occurrence of bluetongue in Northwest- and Central Europe has lead to extensive financial losses. For first-line laboratory diagnosis, reverse transcription real-time PCR (RT-qPCR) has been used increasingly, necessitating careful evaluation of all applied methods by the performing laboratory. In the course of an internal validation, four commercially available BTV RT-qPCR kits and two published reference methods were compared.

Important vectors for Listeria monocytogenes transmission at farm dairies manufacturing fresh sheep and goat cheese from raw milk.

The aim of this study was to determine the transmission routs of Listeria spp. in dairy farms manufacturing fresh cheese made from ovine and caprine raw milk and to evaluate the impact of Listeria monocytogenes mastitis on raw milk contamination. Overall, 5,799 samples, including 835 environmental samples, 230 milk and milk product samples, and 4,734 aseptic half-udder foremilk samples were collected from 53 dairy farms in the dairy intensive area of Lower Austria.

Detection and molecular analysis of West Nile virus infections in birds of prey in the eastern part of Austria in 2008 and 2009.

The emergence of West Nile virus (WNV) was expected in Austria since the initial discovery of the infection in neighbouring Hungary in 2003/2004. In 2008 six cases of West Nile disease were diagnosed at the Institute for Veterinary Disease Control Mödling, Austrian Agency for Health and Food Safety (AGES), involving five goshawks (Accipiter gentilis) and one gyrfalcon (Falco rusticolus), which were found dead in the eastern Austrian federal states of Lower Austria, Vienna and Styria, respectively. Pathomorphological and immunohistochemical findings suggested a WNV infection.

Bluetongue virus RNA detection by RT-qPCR in blood samples of sheep vaccinated with a commercially available inactivated BTV-8 vaccine.

In 2008, a country-wide bluetongue virus 8 (BTV-8) vaccination campaign has been initiated in Austria, using a single commercial inactivated BTV-8 vaccine. Based on preliminary data, we hypothesised that vaccine-derived BTV RNA is transiently detectable by reverse transcription quantitative real-time PCR (RT-qPCR) in the blood of vaccinated animals. Thus BTV-8 vaccine was administered to five BTV-naïve adult sheep and blood samples were taken at various time-points post-vaccination.

Uptake and binding of the serotonin 5-HT1A antagonist [18F]-MPPF in brain of rats: effects of the novel P-glycoprotein inhibitor tariquidar.

We used microPET to map the dose-response to the novel P-glycoprotein (P-gp) inhibitor tariquidar (TQD) of the initial influx of the P-gp substrate [(18)F]-MPPF in rat brain, and to test for effects of P-gp inhibition on the subsequent binding of [(18)F]-MPPF to serotonin 5-HT(1A) receptors. Summation maps of [(18)F]-MPPF uptake during the first 100 seconds after intravenous injection were calculated in groups of rats with vehicle (glucose 5%) pretreatment, or following pretreatment with TQD at doses of 5, 15, or 30 mg/kg. The early summation image (K(1)-weighted), were validated as a surrogate marker for the physiological blood-brain clearance (K(1); ml g(-)(1) min(-1)) by linear graphic analysis of the unidirectional blood-brain clearance relative to an image-based arterial input measured in the left ventricle of the heart.

In vivo down-regulation of mouse brain capillary P-glycoprotein: a preliminary investigation.

Over-expression of blood-brain barrier P-glycoprotein is considered as a major hurdle in the treatment of various CNS disorders. A down-regulation strategy is considered as one means to counteract disease- or therapy-associated induction of P-glycoprotein. Here, we evaluated whether a targeting of P-glycoprotein can be achieved in mouse brain capillary endothelial cells using siRNA.

Outbreak of staphylococcal food intoxication after consumption of pasteurized milk products, June 2007, Austria.

On June 13, 2007, the public health authority informed the Austrian Agency for Health and Food Safety about 40 children from two neighboring elementary schools who had fallen ill with abdominal cramps and vomiting on June 8. School milk products consumed on June 8 were suspected as the source of the outbreak. On June 8, the milk products provided by local dairy X to eight elementary schools and two nurseries.

Tolerability of N-chlorotaurine in the bovine mammary gland.

N-Chlorotaurine (NCT) is a promising endogenous agent for topical treatment of infections. We tested the tolerability and pharmakokinetics of NCT in the bovine mammary glands in a phase 1 study. Three concentrations of NCT in water (0.

Effects of periparturient systemic treatment with penethamate hydriodide on udder health and milk yield of dairy heifers.

Sixty dairy heifers from seven Austrian herds, with high prevalence of Staphylococcus aureus mastitis, were used in this pilot study. Heifers were randomly allocated to two groups. The treatment group received at parturition intramuscularly 10 million i.

The potential of measuring serum amyloid A in individual ewe milk and in farm bulk milk for monitoring udder health on sheep dairy farms.

The aim of the study was to determine the diagnostic value of measuring serum amyloid A (SAA) concentrations in milk of individual ewes and in farm bulk milk for monitoring udder health. Udder health was calculated by examining a randomly selected group of seven flocks at each farm visit by means of California mastitis test and bacteriological examination of 5749 milk samples. SAA was determined additionally in 267 randomly selected milk samples from six flocks.

Cystoscopy in cattle--a valuable additional tool for clinical examination.

Based on the findings of physical examination and on laboratory findings the urinary bladder of 23 cows was examined endoscopically in order to investigate the application of cystoscopy in cattle. The endoscopic findings of all examined cows were compared with the findings of physical examination and the results of macroscopic and microscopic urinalysis and the bacteriological culture of the urine. By physical examination only 3 cows were diagnosed to have urinary tract disease, whereas all other cows were suspected of having an urinary tract disease.

A case of sporadic ovine mastitis caused by Listeria monocytogenes and its effect on contamination of raw milk and raw-milk cheeses produced in the on-farm dairy.

We describe a case of listerial mastitis in a flock of 130 sheep. The animals were housed at a farm where the bulk raw ewe milk was processed to produce raw milk soft cheese. List.

Impact of extracorporeal membrane oxygenation modality on cytokine release during rescue from infant hypoxia.

The treatment of acute respiratory failure in infants by means of extracorporeal membrane oxygenation (ECMO) is thought to be associated with a treatment-related inflammatory reaction, which may deteriorate the underlying disease process. The aim of this study was to compare the venoarterial (VA) and venovenous (VV) modality of ECMO with regard to their pulmonary and serological cytokine release during rescue from acute hypoxia. The inflammatory response was measured in piglets undergoing hypoxic ventilation with a gas mixture of 92% N2 and 8% O2, which were then rescued through VA- (n = 5) or VV-ECMO (n = 5).

Dynamics of experimentally induced Staphylococcus epidermidis mastitis in East Friesian milk ewes.

The responses of five lactating East Friesian milk ewes to experimental mammary infection with Staphylococcus epidermidis and of five control ewes were examined over a period of 10 weeks. Infection caused an influx of neutrophils into milk, the numbers of which started to rise 4 h post infection and peaked 24 h after infection. The initial response was accompanied by mild fever and mild leucopaenia in blood (8 h after infection).

Evaluation of flow injection analysis for determination of urea in sheep's and cow's milk.

Difficulties in measuring the urea content in sheep's milk often occur with spectral photometry due to the high protein and fat concentrations of the milk. In this study an enzymatic flow procedure (QuickChem 8000 Ion Analyser, Lachat Instruments, Milwaukee, USA) to determine the urea content in ovine and bovine milk was evaluated. Urea content is determined by the Berthelot reaction after splitting it enzymatically with urease.