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We investigated the prevalence of Hepatitis E Virus (HEV), Leptospira and Ascaris suum (A. suum) seropositivity, and of nasal methicillin‐resistant Staphylococcus aureus (MRSA) colonization among Austrian practising veterinarians, and assessed the association with occupational swine livestock exposure. The 261 participants completed a questionnaire on demographics, intensity of occupational swine livestock contact and glove use during handling animals and their secretions. Participants' blood samples were tested for HEV, Leptospira and A. suum seropositivity and nasal swabs cultured for MRSA. We compared swine veterinarians (defined as >3 swine livestock visits/week) to non‐swine veterinarians (≤3 swine livestock visits/week) with regard to the outcomes through calculating prevalence ratio (PR) and 95% confidence interval (CI). Furthermore, the relationship between occupational swine livestock contact and the study outcomes was examined by age (3 occupational swine livestock visits per week is associated with HEV and A. suum seropositivity and nasal MRSA colonization and that glove use may play a putative preventive role in acquiring HEV and A. suum. Further analytical epidemiological studies have to prove the causality of these associations.

Zusammenfassung Gegenstand und Ziel Darstellung der Untersuchungsergebnisse von 150 Abortfällen beim Rind aus dem Jahr 2018 (Januar–September). Material und Methoden Je nach Einsendematerial und Anforderung erfolgten serologische, bakteriologische und/oder molekularbiologische Untersuchungen zum Nachweis von in Österreich anzeigepflichtigen und nicht anzeigepflichtigen Aborterregern. Ergebnisse Neben rein tierpathogenen Infektionserregern konnten bei je 1 Tier Brucella melitensis, Salmonella Dublin und bei 2 Feten Coxiella burnetii nachgewiesen werden. Schlussfolgerung Die Ergebnisse zeigen, dass im Zusammenhang mit Aborten immer eine besondere Sorgfaltspflicht hinsichtlich Schutz der exponierten Person (Landwirt, Tierarzt, Laborpersonal) erforderlich ist. Auch mit dem Wiederauftreten von lange nicht mehr aufgetretenen Tierseuchen ist jederzeit zu rechnen, wie aus dem aktuellen Brucella-Nachweis ersichtlich wird. Klinische Relevanz Wichtigste Voraussetzung für eine hohe Aufklärungsrate bei infektiös bedingten Aborten stellt die Wahl des geeigneten Probenmaterials dar, nämlich Fetus mit Plazenta und maternale Blutproben. Aufgrund des großen Erregerspektrums ist die Abortdiagnostik umfangreich und aufwendig.

Mycobacterium caprae subtype Lechtal was detected in a red fox (Vulpes vulpes) shot by a hunter in 2018 in the western part of Austria, where, among wildlife, tuberculosis is known to occur in red deer (Cervus elaphus). The red fox showed a generalized (disseminated) manifestation of the disease and a multibacillary distribution of mycobacteria in the inner organs.

In the study, the laboratory results of 150 bovine abortion cases from 2018 (January-September) are presented.Depending on the submitted sample material and the requested examination, serological, bacteriological and/or molecular biological investigations were performed to detect abortion-causing pathogens which need or do not need to be notified in Austria.In addition to animal pathogens, the zoonotic pathogensThe results show, that because of the zoonotic potential of some pathogens, care must be taken when handling abortion material to ensure that farmers, veterinary surgeons and laboratory staff are not at risk. Taking bovine brucellosis as an example, the reappearance of previously eradicated diseases has to be expected at any time.For detailed diagnostics, fetus with placenta and blood samples from the dam should be submitted to the laboratory. According to the extensive pathogen spectrum, investigation of abortion cases is laborious and time consuming.Darstellung der Untersuchungsergebnisse von 150 Abortfällen beim Rind aus dem Jahr 2018 (Januar–September).Je nach Einsendematerial und Anforderung erfolgten serologische, bakteriologische und/oder molekularbiologische Untersuchungen zum Nachweis von in Österreich anzeigepflichtigen und nicht anzeigepflichtigen Aborterregern.Neben rein tierpathogenen Infektionserregern konnten bei je 1 TierDie Ergebnisse zeigen, dass im Zusammenhang mit Aborten immer eine besondere Sorgfaltspflicht hinsichtlich Schutz der exponierten Person (Landwirt, Tierarzt, Laborpersonal) erforderlich ist. Auch mit dem Wiederauftreten von lange nicht mehr aufgetretenen Tierseuchen ist jederzeit zu rechnen, wie aus dem aktuellenWichtigste Voraussetzung für eine hohe Aufklärungsrate bei infektiös bedingten Aborten stellt die Wahl des geeigneten Probenmaterials dar, nämlich Fetus mit Plazenta und maternale Blutproben. Aufgrund des großen Erregerspektrums ist die Abortdiagnostik umfangreich und aufwendig.

Tick-borne encephalitis virus (TBEV) and West Nile virus (WNV) are important arthropod-borne zoonotic flaviviruses. Due to the emergence of WNV in TBEV-endemic regions co-circulation of both viruses is increasing. Flaviviruses are structurally highly similar, which leads to cross-reacting antibodies upon infection. Currently available serological assays for TBEV and WNV infections are therefore compromised by false-positive results, especially in IgG measurements. In order to discriminate both infections novel diagnostic methods are needed. We describe an ELISA to measure IgG antibodies specific for TBEV and WNV, applicable to human and horse sera. Mutant envelope proteins were generated, that lack conserved parts of the fusion loop domain, a predominant target for cross-reacting antibodies. These were incubated with equine and human sera with known TBEV, WNV or other flavivirus infections. For WNV IgG, specificities and sensitivities were 100% and 87.9%, respectively, for horse sera, and 94.4% and 92.5%, respectively, for human sera. TBEV IgG was detected with specificities and sensitivities of 95% and 96.7%, respectively, in horses, and 98.9% and 100%, respectively, in humans. Specificities increased to 100% by comparing individual samples on both antigens. The antigens could form the basis for serological TBEV- and WNV-assays with improved specificities.

ZusammenfassungGegenstand und Ziel: Das Porzine Reproduktive und Respiratorische Syndrom (PRRS) verursacht weltweit bedeutende wirtschaftliche Verluste in der Schweineproduktion. Die Studie untersuchte den Zusammenhang zwischen dem PRRS-Betriebsstatus, ermittelt durch Nachweis von PRRS-Virus-Antikörpern (PRRSV-Ak) im Blutserum der Schlachtschweine, und den bei den Schlachttierkörpern gefundenen pathologisch-anatomischen Lungenbefunden. Material und Methoden: Zwischen 01.12.2011 und 16.04.2012 wurden an einem österreichischen Schlachthof von 1056 Schlachtschweinen aus 66 schweinehaltenden Betrieben Blutproben entnommen und mittels ELISA auf PRRSV-Ak getestet. In keinem Betrieb fanden Impfungen gegen PRRSV statt. Parallel zur serologischen Untersuchung erfassten amtlich beauftragte Tierärzte am Geschlinge- und Tierkörperband die pathologisch-anatomischen Lungenbefunde aller Schlacht schweine aus diesen Betrieben zwischen 01.09.2011 und 11.12.2012. Die Prävalenz von Lungenentzündungen und Pleuritiden in PRRSV-Ak-positiven und unverdächtigen Beständen wurde verglichen und statistisch ausgewertet. Ergebnisse: Die Prävalenz hochgradiger Lungenentzündungen sowie Pleuritis visceralis und parietalis lag bei Schweinen aus PRRSV-Ak-positiven Betrieben signifikant höher als bei Schweinen aus unverdächtigen Beständen. Dabei ergab sich bei kombinierten Betrieben (Ferkelaufzucht und Mast an einem Standort) eine tendenziell höhere Prävalenz von mittel- und hochgradigen Lungenentzündungen als in reinen Mastbetrieben. Schlussfolgerungen und klinische Relevanz: Der PRRSV-Status von Schweinebetrieben hatte Auswirkungen auf das Auftreten von pathologisch-anatomischen Lungenveränderungen der geschlachteten Schweine. Auch nicht PRRS-typische Befunde wie Pleuritiden traten vermehrt auf. Dies spricht für eine Beteiligung anderer Primär- oder Sekundärerreger, die durch den immunsupprimierenden Effekt des PRRSV begünstigt werden und selbst auch Atemwegserkrankungen auslösen können. In der Tendenz waren dabei die kombinierten Betriebe stärker betroffen als die reinen Mastbetriebe.

Mycobacterium caprae, a member of the Mycobacterium tuberculosis complex, is the main causative agent of bovine tuberculosis in alpine regions. Bacterial culture is the gold standard in bovine tuberculosis diagnostic but takes up to twelve weeks. This increases the time and costs for stocks affected with bovine tuberculosis. Hence this study focused on the implementation of a fast and precise mycobacterial detection method and compared it with currently used methods. Matrix lysis is a chemical lysis using high concentrations of urea to solubilize bovine and red deer tissue and was used to detect even smallest amounts or non-visible lesions of mycobacteria. A total of 64 samples collected from 44 animals (37 red deer and 7 cattle) were tested by Matrix lysis. Forty-three of these samples were used for Mycobacterium tuberculosis complex detection by quantitative PCR and other 21 for subtyping the genetically different variants of M. caprae. Furthermore, three Matrix lysis samples were used for Next Generation Sequencing. Our results confirm that Matrix lysis is a fast and precise method for detecting Mycobacterium tuberculosis complex in native tissue samples. However, at the moment it reaches its limits when the samples were analyzed by Next Generation Sequencing and RD4 subtyping.

Background Effective vaccines against porcine reproductive and respiratory syndrome virus (PRRSV), especially against highly pathogenic (HP) PRRSV are still missing. The objective of this study was to evaluate the protective efficacy of an experimental live attenuated PRRSV 2 vaccine, composed of two strains, against heterologous challenge with a Vietnamese HP PRRSV 2 field strain. For this reason, 20 PRRSV negative piglets were divided into two groups. The pigs of group 1 were vaccinated with the experimental vaccine, group 2 remained unvaccinated. All study piglets received an intranasal challenge of the HP PRRSV 2 on day 0 of the study (42 days after vaccination). Blood samples were taken on days 7 and 21 after vaccination and on several days after challenge. On day 28 after challenge, all piglets were euthanized and pathologically examined. Results On days 7 and 21 after vaccination, a PRRSV 2 viraemia was seen in all piglets of group 1 which remained detectable in seven piglets up to 42 days after vaccination. On day 3 after challenge, all piglets from both groups were positive in PRRSV 2 RT-qPCR. From day 7 onwards, viral load and number of PRRSV 2 positive pigs were lower in group 1 than in group 2. All pigs of group 1 seroconverted after PRRSV 2 vaccination. PRRSV antibodies were detected in serum of all study pigs from both groups from day 14 after challenge onwards. In group 2, moderate respiratory symptoms with occasional coughing were seen following the challenge with HP PRRSV 2. Pigs of group 1 remained clinically unaffected. Interstitial pneumonia was found in four piglets of group 1 and in all ten piglets of group 2. Histopathological findings were more severe in group 2. Conclusions It was thus concluded that the used PRRSV 2 live experimental vaccine provided protection from clinical disease and marked reduction of histopathological findings and viral load in pigs challenged with a Vietnamese HP PRRSV 2 field strain.

This article uses the experience of five European countries to review the integrated approaches (human, animal and vector) for surveillance and monitoring of West Nile virus (WNV) at national and European levels. The epidemiological situation of West Nile fever in Europe is heterogeneous. No model of surveillance and monitoring fits all, hence this article merely encourages countries to implement the integrated approach that meets their needs. Integration of surveillance and monitoring activities conducted by the public health authorities, the animal health authorities and the authorities in charge of vector surveillance and control should improve efficiency and save resources by implementing targeted measures. The creation of a formal interagency working group is identified as a crucial step towards integration. Blood safety is a key incentive for public health authorities to allocate sufficient resources for WNV surveillance, while the facts that an effective vaccine is available for horses and that most infected animals remain asymptomatic make the disease a lesser priority for animal health authorities. The examples described here can support other European countries wishing to strengthen their WNV surveillance or preparedness, and also serve as a model for surveillance and monitoring of other (vector-borne) zoonotic infections.

Schmallenberg virus (SBV) has emerged in summer-autumn 2011 in north-western Europe. Since then, SBV has been continuously spreading over Europe, including Austria, where antibodies to SBV, as well as SBV genome, were first detected in autumn 2012. This study was performed to demonstrate the dynamics of SBV spread within Austria, after its probable first introduction in summer 2012. True seroprevalence estimates for cattle and small ruminates were calculated to demonstrate temporal and regional differences of infection. Furthermore, the probability of SBV genome detection in foetal tissues of aborted or stillborn cattle and small ruminants as well as in allantoic fluid samples from cows with early foetal losses was retrospectively assessed. SBV first reached Austria most likely in July-August 2012, as indicated by retrospective detection of SBV antibodies and SBV genome in archived samples. From August to October 2012, a rapid increase in seroprevalence to over 98% in cattle and a contemporaneous peak in the detection of SBV genome in foetal tissues and allantoic fluid samples was noted, indicating widespread acute infections. Notably, foetal malformations were absent in RT-qPCR positive foetuses at this time of the epidemic. SBV spread within Austrian cattle reached a plateau phase as early as October 2012, without significant regional differences in SBV seroprevalence (98.4-100%). Estimated true seroprevalences among small ruminates were comparatively lower than in cattle and regionally different (58.3-95.6% in October 2012), potentially indicating an eastward spread of the infection, as well as different infection dynamics between cattle and small ruminants. Additionally, the probability of SBV genome detection over time differed significantly between small ruminant and cattle samples subjected to RT-qPCR testing.

Toxoplasma gondii is the aetiological agent of the zoonotic disease toxoplasmosis and transmitted among other ways by chemically and physically untreated, that is, raw pork to humans. The detection of Toxoplasma gondii is impossible by currently practiced meat inspection, but serological tests can be used to detect Toxoplasma gondii antibodies in pig herds and can consequently be helpful to identify potentially contaminated pork. Therefore, appropriate serological tests are required. In this study, serum samples of 1368 naturally exposed slaughter pigs from 73 Austrian farms were collected. Serum samples of at least 16 slaughter pigs per farm were tested. The prevalence of Toxoplasma gondii antibodies in serum was measured by a commercial available modified agglutination test (MAT) and compared to three different commercial available enzyme-linked immunosorbent assays (ELISA). The MAT detected 6.5%, ELISA I 6.7%, ELISA II 4.8% and ELISA III 4.3% of the pigs as Toxoplasma gondii antibody positive. The agreement, according to the kappa coefficient (κ), was substantial between the MAT and ELISA I (κ = 0.62), II (κ = 0.64) and III (κ = 0.67). A better agreement was determined between ELISA I and II (κ = 0.715), ELISA I and III (κ = 0.747) and ELISA II and III (κ = 0.865). At least one pig per farm was detected Toxoplasma gondii antibody positive in 17 (23.3%) farms by the MAT, 26 (35.6%) farms by ELISA I, 16 (21.9%) farms by ELISA II and 11 (15.1%) farms by ELISA III. Pig farms with a high number of Toxoplasma gondii antibody-positive pigs or high antibody titres were identified by all of the four used serological tests. Concerning the occurrence of Toxoplasma gondii antibodies in Austrian pig farms, a monitoring and surveillance programme would be reasonable to find high-risk farms.

The effectivity of different sampling schemes for the early detection of the introduction of porcine reproductive and respiratory syndrome virus into a pig herd was evaluated using Monte Carlo simulation. Within a theoretical breeding herd of 300 animals, disease transmission was simulated using a stochastic SEIR model incorporating actual animal movement data. The following parameters were evaluated for different sample sizes, sampling frequencies and diagnostic procedures (ELISA, PCR): the time from virus introduction until detection, the daily detection probability and the number of holdings to which infected animals are shipped before the disease is detected. The results show that the sample size has an influence on early detection. The biggest effects are, however, achieved by shortening the sampling intervals. The median detection time is approximately ten days shorter for PCR than for ELISA. If, however, the sampling intervals are chosen too wide there is a chance of overlooking the disease using PCR alone.

A retrospective epidemiological investigation of molecular and animal husbandry data collected over an observation period of five years (2009-2014) within the compulsory bovine viral diarrhoea virus (BVDV) control programme in Western Austria, covering the federal provinces of Tyrol and Vorarlberg is presented in this study. Samples collected from 232 infected calves were phylogenetically classified based on the 5' untranslated region (5'UTR). All but 13 samples, which were typed as border disease virus subtype 3 (BDV-3), belonged to the bovine viral diarrhoea virus genotype 1 (BVDV-1) and clustered within six different subtypes (1b, 1e, 1f, 1h, 1d and 1k). Movement data and survival times from infected individual animals were analysed because of their potential of passing on infection to naive herds. From the moment of submission of the laboratory results, 180 animals were culled within the first month, 13 lived longer than two but not longer than six months and seven infected animals lived longer than one year. 13 of the infected animals were born on alpine pastures and eleven infected animals were grazed on mountain pastures during summer. The movement of infected animals and the role of trade in alpine areas are a possible source for spreading the infection, thus hampering the progress of eradication.

Aim of the study was to detect antibodies and potential risk factors for an infec- tion with Leptospira in horses in Middle Germany. Serum samples of 314 horses were examined retrospectively by microscopic agglutination test for the presence of antibodies against eight Leptospira serovars. In total, 17.2% (n = 54) of the horses were positive for one or more of the serovars analyzed. The most prevalent serovar was lcterohaemorrhagiae (11.1%), followed by serovar Bratislava (9.6 %) and Grippotyphosa (1.9%). Mares showed a significantly higher occurrence of antibodies (p0.05) than geldings or stallions. Horses used for breeding have a significantly lower risk than horses used in sport or horses used for leisure activity. There was also a significantly higher prevalence (p0.05) in summer than in the other seasons. No significant influence of breed, husbandry conditions and age on the antibody occurrence was observed (p0.05). The clinical chemical parameters did not differ significantly between horses with positive or negative Leptospira antibody result (p0.05). It became apparent that horses can be infected with Leptospira without developing of clinical symptoms.

A survey of 143 hunter-harvested red deer for tuberculosis was conducted in an Alpine area in Western Austria over two subsequent years. There, single tuberculosis cases caused by Mycobacterium caprae had been detected in cattle and red deer over the preceding decade. The area under investigation covered approximately 500 km2, divided into five different hunting plots. Lymph nodes of red deer were examined grossly and microscopically for typical tuberculosis-like lesions and additionally by microbiological culturing. Executing a detailed hunting plan, nine M. caprae isolates were obtained. Six out of nine originated from one single hunting plot with the highest estimated prevalence of tuberculosis, that is, 23.1%. All isolates were genotyped by mycobacterial interspersed repetitive unit—variable number of tandem repeat (MIRU-VNTR) typing of 24 standard loci plus VNTR 1982. All nine isolates belonged to a single cluster termed “Lechtal” which had been found in cattle and red deer in the region, demonstrating a remarkable dominance and stability over ten years. This is the first report on a systematic prospective study investigating the prevalence and strain variability of M. caprae infection in red deer in Austria and in the Alpine countries.

Aujeszky's disease (AD), caused by Suid herpesvirus type 1 (SuHV-1), is an economically important disease in domestic swine. Thus, rigorous control programmes have been implemented and consecutively AD in domestic swine was successfully eradicated in many countries, including Austria. However, SuHV-1 continues to thrive in wild boar populations, as indicated by high seroprevalences in a number of European countries and by occasional cases of AD in hunting dogs. For the first time, SuHV-1 was detected in Austrian wild boar and a molecular characterization of SuHV-1 isolated from wild boar and hunting dogs was performed. Results of preliminary serological analyses suggest a regional SuHV-1 seroprevalence of over 30% in free-living and almost 70% in fenced wild boar from Eastern Austria. Molecular typing of Austrian SuHV-1 isolates of wild boar origin revealed the presence of two genetically distinct variants of SuHV-1, both capable of infecting dogs that have been exposed to infected wild boar during hunting.

In den meisten europaischen Landern wird die chirurgische Kastration der mannlichen Ferkel routinemasig zur Vermeidung von Ebergeruch durchgefuhrt. Als tierschutzgerechte Alternative steht die Impfung der mannlichen Mastschweine mit dem Gonadotropin-releasing-Hormon (GnRH)-Analogon Improvac® zur Verfugung. Ein wichtiger Aspekt bei der Diskussion von jeglichen Alternativen zur bisherigen Kastrationspraxis liegt in der Akzeptanz der jeweiligen Methode beim Verbraucher. Ziel der Studie war es, in der deutschen Bevolkerung den Kenntnisstand bezuglich Ebergeruchs, chirurgischer Kastration sowie Impfung mannlicher Mastschweine mit Improvac® zu ermitteln sowie die von den Verbrauchern bevorzugte Methode zur Vermeidung von Ebergeruch. Dazu wurden im Oktober 2010 gemeinsam mit dem Institut fur Meinungsforschung Allensbach eine Befragung von 1.786 Personen durch 420 Interviewer durchgefuhrt. Der uberwiegende Teil der Befragten (63 %) kannte keinen Ebergeruch. Nur wenige (24 %) hatten von der chirurgischen Kastration zur Vermeidung von Ebergeruch gehort, nur 6 % kannten die Impfung mit Improvac®. Nach Information uber beide Methoden bevorzugten 41 % der Befragten die Impfung gegenuber 19 %, die die chirurgische Kastration bevorzugten. Interessanterweise konnten sich 40 % der Befragten nicht fur eine der beiden Methoden entscheiden. Es kann geschlussfolgert werden, dass die Impfung gegen Ebergeruch fur die Verbraucher akzeptabel ist, wenn sie uber das Thema korrekt informiert wurden.

Vaccination of male fattening pigs with a gonadotropin releasing factor (GnRF) vaccine is regarded as a possible solution to solve the welfare problem associated with surgical castration, which causes pain and stress even when performed under local or general anaesthesia. The objective of the present study was to compare the behaviour of male fattening pigs either surgically castrated without anaesthesia (T1) or vaccinated twice with a GnRF vaccine (T2). Data collection took place in a commercial German fattening unit. Each treatment comprised 8 groups of 12 pigs, housed in fattening pens with partially slatted floor and liquid feed provided three times a day. Data on postures were scored from 24-h videos recorded in every week of the fattening period (16 weeks) using scan sampling with 5min intervals. Social behaviour was analysed in weeks 2, 4, 6, 8, 10, 12, 14, 15 and 16 by continuous behaviour recording of focus animals in four blocks of 2h phased evenly during the day. Overall, during the whole fattening period, vaccinates (T2) were more active than surgical castrates (T1), indicated by a higher proportion of pigs standing (T1: 9.3%; T2: 10.74%; P P =0.064) and on biting and fighting ( P =0.151) were found. In T2 the prevalence of aggressive behaviours decreased after the second vaccination ( P

Background The aim of the study was to evaluate the performance of different newly developed and/or commercially available ELISAs for detection of PRRSV specific antibodies. Consequently, ten PRRSV negative piglets (group V) were vaccinated with a PRRSV type 2 vaccine. Blood samples were taken before as well as seven, 21 and 42 days after vaccination. At day 42 after vaccination (day 0 of the study) all of the piglets from group V and 10 non-prevaccinated PRRSV negative piglets (group N) were challenged with an HP PRRSV type 2 field strain. Blood samples were taken before and at days 3, 7, 10, 14, 21 and 28 after challenge. The success of vaccination and challenge was measured with RT qPCR. All serum samples were tested with six ELISAs for detection of PRRSV antibodies. Three of them are nucleocapsid-based, two use a glycoprotein extract and one uses inactivated whole virus as antigen. The specificity of the ELISAs was evaluated using 301 serum samples of piglets from PRRSV negative herds. Results The piglets from group V tested positive by RT qPCR at day 7 after vaccination and all piglets tested positive at day 3 after challenge. PRRSV specific antibodies were seen with all nucleocapsid-based ELISAs from day 21 after vaccination onwards in group V and from day 10 after challenge in group N. The glycoprotein-based ELISAs detected antibodies from day 42 after vaccination (group V) and day 21 after challenge (group N). The agreement according to kappa-coefficient was almost perfect. The glycoprotein-based ELISAs were able to distinguish PRRSV type 2, although with some cross reactions. Regarding specificity, the ELISAs performed differently (specificity between 97.4 % and 100 %), whereas most of the ELISAs with higher sensitivity had a slightly lower specificity. Conclusions All tested ELISA were able to detect PRRSV antibodies in the serum of pigs vaccinated with a PRRSV type 2 vaccine and after challenge with an HP PRRSV type 2 field strain. The onset on antibody detection differed, depending on the type of antigen used in the ELISAs. Most of the ELISAs with a higher sensitivity had a lower specificity.

The health status of a 4-year-old female, dd-haplotype miniature pig deteriorated rapidly, so the animal finally had to be euthanized because of poor clinical condition. Necropsy revealed a massive leukocytic infiltration in the parenchymatous organs of the abdominal cavity. on hematologic cell counting, severe leukocytosis (69.3 X 109cells/liter) and high-grade basophilia (6.9 X 109cells/liter) were evident. cytologic examination, as well as analysis of expression of leukocyte differentiation antigens by means of flow cytometry, classified blasts, which accounted for about 22% of leukocytes, as biphenotypic cells co-expressing the myeloid marker SWC3 (CD172a) and the lymphoid markers CD5 and CD25. Hematologic features resembled those seen in humans with chronic myeloid leukemia at blast phase.

The biologically active form of vitamine D(3) [1alpha,25(OH)(2)D(3)] has recently been described not only to influence bone metabolism but also to exert immunomodulating activities, which may have an impact on bone formation/resorption as well. In this study, we analysed the effects of 1alpha,25(OH)(2)D(3) on the cytokine pattern of porcine bone marrow-derived cells from piglets aged 1-3 weeks. After culture for 1 week, the number of osteoclasts was determined, with tartrate-resistant acid phosphatase (TRAP)-positive, multinucleated cells being considered osteoclasts. Cultured bone marrow cell-derived mRNA was subjected to semiquantitative RT-PCR specific for a panel of porcine cytokines (IL-1alpha, IL-6, IL-8, IL-10, and TNF-alpha). In addition, an immunofluorescence analysis using anti-porcine mAbs specific for IL-1beta, IL-2, IL-4, IL-6, IL-12, TNF-alpha, and IFN-gamma was performed. In order to prove the existence of a porcine homologue of the receptor activator of NF-kappaB ligand (RANKL) bone marrow cell- as well as porcine white blood cell-derived mRNA was investigated by RT-PCR using primer pairs specific for murine RANKL. Cell culture supernatant was analysed for soluble RANKL by means of an ELISA designed for quantification of human RANKL. By means of RT-PCR, expression of IL-1alpha, IL-6, IL-8, IL-10 and TNF-alpha mRNA could be found in cells cultured with and without 1alpha,25(OH)(2)D(3). Immunofluorescence analysis revealed that IL-1, IL-6, and TNF-alpha were produced by both stromal cells and osteoclasts. Besides its known osteoclastogenic effects, 1alpha,25(OH)(2)D(3) tended to downregulate the respective cytokines, but significantly upregulated RANKL expression. The homology between the porcine RANKL-specific sequence and the corresponding human RANKL sequence was 79%. The data found support the idea that porcine bone marrow cell cultures may provide a suitable alternative to murine systems in human osteological research.

Porcine reproductive and respiratory syndrome virus is a virus with a relatively high level of sequence variation, especially between the European and the American-type strains. This high degree of sequence variations will influence the accuracy of results obtained by molecular diagnostic methods, especially if mutations are located in the primer or probe binding regions of PCR or real-time PCR based assays. In order to provide the basis for the development of an accurate, region-tailored assay we have analysed samples obtained from several Austrian pig herds using a series of RT-PCR assays. Subsequent sequencing of the PCR products from ORF5 and phylogenetic analysis revealed the occurrence of both the European and the American-type of porcine reproductive and respiratory syndrome virus in Austria. Since vaccination with the American-type vaccine is not authorized in Austria the import of vaccinated animals is the most likely explanation for the occurrence of these strains. The study highlights the importance of a detailed study on the sequence variations occurring in the region of interest before the development of a reliable PCR-based assay.

Summary The thyroglobulin of a ram of the East Friesian milk sheep breed suffering from goitre was investigated by physico- and immunochemical methods. The respective ram was the only animal amongst the other sheep of the flock, that exhibited severe goitre, additionally showing depressed behaviour. Results of the thyroid-stimulating hormone response test were indicative of hypothyroidism. The dysfunction of the thyroid gland could be treated by additional iodine supplementation quite successfully, although all sheep had been given iodinated cattle salt throughout the course of the history. Without reducing conditions sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE) of isolated thyroglobulin molecules of the ram and control sheep did not reveal different band patterns, but under reducing conditions different band patterns were evident for the respective animals: the ram's thyroglobulin displayed two main bands, those of healthy reference sheep only one. Both bands reacted equally with anti-thyroglobulin antibodies, even with those produced by immunizing rabbits with single bands. The reduced single thyroglobulin band of healthy sheep corresponded to a truncated form of that molecule, whereas the additional main band of the ram was a more resistent, intact thyroglobulin subunit, as was shown by mass spectrometry. In conclusion, results of physico- and immunochemical investigations gave evidence of a modification of thyroglobulin with suspected different iodine binding properties in the ram. The latter finding may have clinical relevance in similar cases in other species, as it is an example of the impact that a minor change in a protein molecule may have on a complete metabolic pathway. Additionally, it could be shown, that in the ovine species the generally found single main band of thyroglobulin after reduction is a truncated form and not an intact subunit. This truncation seems to be induced in vitro by the reductive sample pretreatment prior to SDS-PAGE.

Summary The nomenclature of ulcerative dermatitis as used in literature is somehow confusing because on the one hand this skin disorder is associated with bacterial growth and on the other hand it is a synonym for a chronic sporadic disease of adult sows with unknown aetiology. Thus, we propose the terminus ‘Porcine Ulcerative Dermatitis Syndrome (PUDS)’ for the latter to distinguish between these two disease complexes. This syndrome could be identified by clinical and pathological examinations in six sows, that were submitted to the clinic. Epidermal ulcers could be found nearly all over the body, but teats were always spared. Haematological examination showed a slight anaemia but physiological leucocyte counts. However, lymphopenia (x = 44.8%), granulocytosis (x = 42.0%) and an increased number of monocytes (x = 13.1%) could be found. Histologically, a lymphoplasmacytic and granulohistiocytic infiltration in the corium was most prominent. In some cases, a moderate leucocytoclastic vasculitis and perivasculitis could be seen at the dermo-epidermal border. Additionally, a multifocal interstitial nephritis with lymphoplasmacytic infiltration was a prominent feature in all animals. Participation of an immune complex associated disorder can be assumed when regarding histological findings as skin lesions in combination with glomerulonephritis are a common feature of such diseases. Also, IgG levels were elevated two- to fourfold in all affected sows when compared with healthy control pigs. This supports the hypothesis that not only T cells, as shown previously, but also the humoral branch of the immune system is involved in the aetiology of PUDS.

In the present study, ribosomal RNA (rRNA) gene activation, monitored through nucleolus development, was studied by autoradiography following 3H-uridine incubation, transmission electron microscopy, and immunofluorescence confocal laser scanning microscopy of key nucleolar proteins involved in rRNA transcription (topoisomerase I, upstream binding factor, and RNA polymerase I) and processing (fibrillarin, nucleolin, and nucleophosmin) in in vivo developed, in vitro produced, and parthenogenetic bovine embryos. In general, in vivo developed embryos displayed formation of fibrillo-granular nucleoli during the 4th post-fertilization cell cycle. During the previous stages of development, nucleolus precursor bodies (NPBs) were observed. However, on some occasions the initial steps of nucleolus formation were observed already at the 2- and 4-cell stage in cases where such embryos were collected from superovulated animals together with later embryonic stages presenting nucleolar development and autoradiographic labeling. The in vitro produced embryos displayed very synchronous formation of fibrillo-granular nucleoli and autoradiographic labeling during the 4th cell cycle. In vivo developed and in vitro produced embryos displayed allocation of nucleolar proteins to fibrillar and granular compartments of the developing nucleoli during the 4th cell cycle. The parthenogenetic embryos typically displayed formation of fibrillo- granular nucleoli during the 5th cell cycle and autoradiographic labeling was not observed until the morula stage. Moreover, the 1-, 2-, and 4-cell parthenogenetic embryos practically lacked NPBs. On the other hand, parthenogenetic embryos displayed allocation of nucleoar proteins to nuclear entities during the 4th cell cycle. In conclusion, both in vivo developed and in vitro produced bovine embryos displayed activation of transcription and nucleolar development during the 4th cell cycle. However, in vivo developed embryos flushed together with later developmental stages displayed premature activation of these processes. Parthenogenetic bovine embryos, on the other hand, displayed a delayed activation. Mol. Reprod. Dev. 65: 73–85, 2003. © 2003 Wiley-Liss, Inc.

Objective of the study was to evaluate the influence of time point of second vaccination with the GnRH analogon Improvac on growth performance, carcass quality and fatty acid composition of male fatteners compared to surgically castrated pigs and entire boars. The pigs (Piétrain-crossbreds) were divided into two vaccination groups with first GnRH vaccination at eleven weeks of age and second vaccination at 21 (group IA, n = 84) or 18 weeks (IB, n = 83) of age, one group with surgically castrated males (C, n = 90) and one with entire males (EM, n = 91). Body weight, feed conversion rate, carcass quality and fatty acid composition in back fat were estimated. Feed conversion rate until second vaccination was better (P0.05) in the vaccination groups (1:2.39) and in group EM (1:2.34) than in group C (1:2.55). Carcass weight did not differ between the groups. Vaccination groups had significantly (P0.01) leaner meat (IA: 58.9%, IB: 58.3%) and less back fat (IA: 14.6 mm, IB: 15.5 mm) than group C (56.5%, 17.1 mm). Fatty acid composition was shifted to polyunsaturated fatty acids (PUFA) in back fat in vaccination groups and EM compared to C. The time lag between second vaccination and slaughter had no influence on growth performance, feed intake and carcass quality. C18:3 and C20:2 were significantly (P0.01) higher in group IB than in IA, but PUFA did not differ between vaccination groups. GnRH vaccinated fatteners were economically superior to surgically castrated in this study.

Background In recent years, several new ELISAs for the detection of antibodies against the porcine reproductive and respiratory disease virus (PRRSV) in pig serum have been developed. To interpret the results, specificity and sensitivity data as well as agreement to a reference ELISA must be available. In this study, three commercial ELISAs (INgezim PRRS 2.0 - ELISA II, Priocheck® PRRSV Ab porcine – ELISA III and CIVTEST suis PRRS E/S PLUS - ELISA IV, detecting PRRSV type 1 antibodies) were compared to a standard ELISA (IDEXX PRRS X3 Ab Test - ELISA I). The serum of three pigs vaccinated with an attenuated PRRSV live vaccine (genotype 2) was tested prior to and several times after the vaccination. Furthermore, serum samples of 245 pigs of PRRSV positive herds, 309 pigs of monitored PRRSV negative herds, 256 fatteners of assumed PRRSV negative herds with unknown herd history and 92 wild boars were tested with all four ELISAs. Results ELISAs II and III were able to detect seroconversion of vaccinated pigs with a similar reliability. According to kappa coefficient, the results showed an almost perfect agreement between ELISA I as reference and ELISA II and III (kappa > 0.8), and substantial agreement between ELISA I and ELISA IV (kappa = 0.71). Sensitivity of ELISA II, III and IV was 96.0%, 100% and 91.5%, respectively. The specificity of the ELISAs determined in samples of monitored PRRSV negative herds was 99.0%, 95.1% and 96.4%, respectively. In assumed negative farms that were not continually monitored, more positive samples were found with ELISA II to IV. The reference ELISA I had a specificity of 100% in this study. Conclusions All tested ELISAs were able to detect a PRRSV positive herd. The specificity and sensitivity of the tested commercial ELISAs, however, differed. ELISA II had the highest specificity and ELISA III had the highest sensitivity in comparison to the reference ELISA. ELISA IV had a lower sensitivity and specificity than the other ELISAs.

Recently, research into alternative sample materials, such as oral fluid or filter-dried blood has been intensified, in order to facilitate cost-effective and animal-friendly sampling of individuals or groups of pigs for diagnostic purposes. The objective of this study was to compare the sensitivity of porcine reproductive and respiratory syndrome virus (PRRSV)-RNA detection by reverse transcription quantitative real-time PCR (RT-qPCR) in serum, FTA filter-dried blood and oral fluid sampled from individual pigs. Ten PRRSV negative pigs were injected with an EU-type PRRSV live vaccine. Blood and oral fluid samples were taken from each pig before, and 4, 7, 14 and 21 days after vaccination. All samples were then analyzed by PRRSV RT-qPCR. In serum, eight often pigs tested RT-qPCR positive at different time points post infection. Absolute quantification showed low serum PRRSV-RNA loads in most samples. In comparison to serum, sensitivity of PRRSV-RNA detection was strongly reduced in matched FTA filter-dried blood and in oral fluid from the same pigs. These results indicate that with low PRRSV-RNA loads the diagnostic sensitivity of PRRSV-RNA detection by RT-qPCR achieved with serum is currently unmatched by either FTA filter-dried blood or oral fluid.

A sperm cytosolic factor is responsible for oocyte activation at fertilization in mammals. The molecular identity of this factor is not yet known, although a sperm phospholipase Cgamma (PLCgamma) is a potential candidate. In this study, cation-exchange chromatography with a Heparin column was used for the fractionation of porcine sperm cytosolic extracts. Oocyte activation potential of the resulting fractions was tested and active fractions were subjected to Western blot analysis using antibodies specific to PLCgamma1. PLCgamma1 was detected in fractions other than those supporting oocyte activation (Ca(2+)-release and pronuclear formation). The active Heparin fraction was then purified on a Mono Q anion-exchange column. One of the resulting fractions still contained Ca(2+)-releasing activity, but pronuclear formation did not occur. We conclude that sperm PLCgamma1 is not involved in oocyte activation and that Ca(2+)-release and pronuclear formation requires multiple factors from sperm cytosol.

In order to identify ESTs which represent possible candidates for carcass traits in pigs, the differential display approach was used. F2 animals of a resource population and pure-bred German Landrace (DL) pigs were selected for the trait ‘eye muscle area’ in order to build up groups of three high and three low performing individuals within each population. To increase the probability that differentially expressed DNA fragments were not found due to the genetic background but due to differences in a few genes affecting the trait of interest, siblings were included in the high and in the low performing groups. RNA was isolated from M. longissimus dorsi and four ‘intra-litter constrasting pools’ were prepared: high performing F2, low performing F2, high performing DL and low performing DL. Differential display banding patterns were produced using (d)T11VA (V:A,C,G) and 20 arbitrary primers. Comparing the banding patterns of the four RNA pools revealed 27 nonshared bands. Here we report on the analysis of seven of these bands, including sequencing, search for homology and mapping using a somatic cell hybrid panel. Two clones showed high homology to known genes, two were homologous to an EST and a SINE sequence. Three clones did not show any homology. Differential expression was tested by semiquantitative reverse transcription–polymerase chain reaction (RT–PCR) and could be confirmed for six clones.

This case report is the first description of a fibroepithelial hamartoma in a pig. The dysplasia, which covered half of the face of the newborn piglet, did not increase in relative size until the animal was euthanatized at 6 months of age. Histologic examination revealed a moderate orthokeratotic hyperkeratosis with variable degrees of epithelial proliferation. The main body of the dysplasia consisted of collagenous fibers. In addition, some proliferating small blood vessels as well as focally gathered dilated apocrine glands were evident. Given morphologic and clinical features, the diagnosis of a hamartoma seemed to be justified.

BackgroundLeptospirosis is one of the world's mostly spread zoonoses causing acute fever. Over years, leptospirosis has been reported to occur rarely in Austria and Germany (annual incidence of 0.06/100,000 in Germany). Only imported cases have been on the increase. Objectives of this case-series study were to retrospectively assess epidemiologic and clinical characteristics of leptospirosis illnesses in South-East Austria, to describe risk exposures for autochthonous infections, and to compare patients with imported versus autochthonous infection.Methodology/principal findingsDuring the 9-year period between 2004 and 2012, 127 adult patients (49 females, 78 males) who tested positive by rapid point-of-care test for Leptospira-specific IgM (Leptocheck®) were identified through electronic hospital databases. Follow-up telephone interviews were conducted with 82 patients. A total of 114 (89.8%) of the 127 patients enrolled had acquired leptospirosis within Austria and 13 (10.2%) had potentially imported infections. Most autochthonous cases were diagnosed during the months of June and July, whereas fewest were diagnosed during the winter months. Exposure to rodents, recreational activities in woods or wet areas, gardening, cleaning of basements or huts were the most common risk exposures found in autochthonous infection. Serogroups Australis (n = 23), Sejroe (n = 22), and Icterohaemorrhagiae (n = 11) were identified most frequently by MAT testing in autochthonous infections. Patients with imported leptospirosis were significantly younger, less likely to be icteric and had significantly lower liver transaminase levels (p = 0.004) than those with autochthonous infections.Conclusions/significanceLeptospirosis is endemic in South-East Austria. In contrast to reports from other countries we found a relatively high proportion of leptospirosis cases to be female (39% vs. ∼ 10%), likely the result of differing risk exposures for South-East Austria.

Traditionally, quantitative microbial risk assessment (QMRA) is based on culture-dependent technologies. However, molecular quantification could forge additional, detailed information. A prerequisite of quantitative real-time PCR in animal science is a tissue preparation method where large volumes of tissue material can be reduced and particularly target cells can be concentrated. An easy-to-use sample preparation method for food (Matrix-Lysis) was recently adapted to tissues and now permits quantification of target cells from up to 5 g of organic matrix. The aim of this study was to examine the suitability of Matrix-Lysis for quantification of Salmonella in porcine ileocaecal lymph nodes (ICLNs). After demonstrating constant recovery rates, ICLNs from 540 pigs were examined for Salmonella spp. with Matrix-Lysis. Samples were also analysed using ISO 6579:2002, a combined enrichment/qPCR method and a lateral flow test. It could be shown that qPCR coupled with Matrix-Lysis can contribute to QMRA in food safety by enabling reproducible quantitative data, even at low contamination rates.

Granulosa-cumulus and cumulus-corona expansion as well as aromatase localization within ovarian follicles were monitored during the preovulatory period in superovulated cattle that were blood sampled every 2'nd h for LH analyses. Granulosa-cumulus as well as cumulus-corona expansion were studied by means of transmission electron microscopy and computerized image analysis. Localization of aromatase, an enzyme involved in estrogen synthesis, was determined immunocytochemically using anti-human placental aromatase cytochrome P-450 antisera. Nuclear oocyte maturation was determined by aceto:orcein staining. Significant cell dissociation within the granulosa-cumulus stalk occurred before the breakdown of the germinal vesicle, i.e. the oocyte nucleus, during the period up to 5-7 h after the LH peak, i.e. the highest LH concentration during the surge. Significant increase in intercellular spacing between the cumulus-corona cells occurred at 13-15 and 19-21 h after the LH peak. Before the LH peak all layers of granulosa cells were immunocytochemically stained for aromatase. At 5-7 h after the LH peak, however, only the granulosa cell layers located near the basal lamina were stained, and at all later intervals staining was absent. The granulosa cells of primary and secondary follicles, the interstitial gland cells, the theca interna cells and the oocytes in all follicles were immunocytochemically unstained.

Summary Apa I and Cfo I polymorphisms in the fragment spanning from the first exon to the third intron of the growth-hormone gene were analysed in 77 Osterreichischen Edelschwein and 269 Osterreichischen Landrasse pigs. The interrelationships between them and the Hae II, Msp I, and ryr 1 polymorphisms were investigated. The differences in distribution between the two breeds were significant at all loci (p < 0.05 to P < 0.01). It is concluded that the Apa I, Cfo I, Hae II, and Msp I pGH polymorphisms analysed can be used as genetic markers for the breeds. The interactions between the analysed polymorphic systems vary considerably. The strongest interaction was found between Cfo I and Hae II in both breeds (C = 0.85). This might be a result of similarity in their restriction enzyme sites. No interactions could be found between the pGH and the ryr 1 locus. Zusammenfassung Apa I- und Cfo I-Polymorphismen des porcinen Wachstumshormongens (pGH-Gen) bei zwei ryr 1-genotypisierten, osterreichischen Schweinerassen An 77 Tieren des Osterreichischen Edelschweins und 269 Tieren der Osterreichischen Landrasse wurden die Verteilungen des Apa I- und des Cfo I-Polymorphismus des porcinen Wachstumshormongens ermittelt, und ihre Beziehungen zu den Hae II- und Msp I-Polymorphismen am selben Genom, sowie zum MHS-Locus untersucht. Die Verteilungsunterschiede zwischen des beiden Rassen war bei Ana I mit p < 0,05 statistisch signifikant, bei den anderen Polymorphismen mit p < 0,01 statistisch hoch signifikant. Daraus last sich eine gute Eignung von Apa I, Cfo I, Hac II und Msp I als genetische Marker fur die untersuchten Rassen ableiten. Die Zusammenhange zwischen des polymorphen Systemen sind zum Teil sehr unterschiedlich ausgepragt. Den hochsten Beziehungsgrad konnte bei beiden Rassen zwischen Cfo 1 und Hae II nachgewiesen werden (C = 0,85), was auf die Ahnlichkeit ihrer Restriktionsenzym-Erkennungs-Sequenzen zuruckgefuhrt werden kann. Zwischen dcm pGH- und dcm ryr 1-Locus konnten keine nennenswerten Interaktionen festgestellt werden.

Brucella canis occurs almost worldwide and is a potential danger to the health of dogs and humans. The pathogen was detected in the placenta and fetuses of a Standard Poodle by direct culture and immunohistochemistry. Further, Brucellae were also isolated from the blood samples of two asymptomatic female Medium Poodles. The isolates were identified as B. canis by conventional microbiological methods and a novel Bruce-ladder multiplex PCR. Genotyping was performed by multiple locus variable number tandem repeats analysis (MLVA).

Endoscopic submucosal dissection (ESD) demands a new level of endoscopic skill in Europe. A 2-day workshop was set up for trainees to carry out five ESD each in order to obtain the skill level required to perform ESD in the stomach or rectum. This study describes: (i) the workshop setup; (ii) the participant's performance; and (iii) the training effect on post-workshop clinical ESD performance.Eighteen very experienced European endoscopists participated in four half-day (4.5 h) training sessions, with everybody rotating daily through six separate training stations (two each with dual, hook, or hybrid knives) with expert tutors. One anesthetized piglet was used per station and session. After 1 year, the clinical ESD performance was surveyed to estimate the training effect of the workshop.Overall, 74 ESD were performed, that is, 4.1 ESD per participant. On average ESD lasted 57 min for 6 cm(2) specimens. We detected a 22% rate of perforation (16 of 74 ESD with perforations), mostly attributable to participants with less experience in ESD. Those who started clinical ESD within 1 year after the workshop performed 144 clinical ESD (median 8 [0-20] per trainee) mostly in the stomach (40%) and large bowel (46%) with an acceptable rate of perforation (9.7%) and surgical repair (3.5%) without mortality or persistent morbidity.Intense skill training for ESD is needed to reduce the risk of perforation, as demonstrated by the results of this workshop. We show that experimental ESD training, however, enables skilled European endoscopists to perform ESD in standard locations with moderate risk of perforation during the clinical learning curve.

Thirty-eight natural cases of aetiologically unclear non-suppurative encephalitis in pigs were studied retrospectively. Brain samples were examined for the presence of porcine circovirus type 2 (PCV-2), porcine respiratory and reproductive syndrome virus (PRRSV), porcine enteroviruses (PEVs), ovine herpesvirus type 2 (OvHV-2), Borna disease virus (BDV) and suid herpesvirus type 1 (SuHV-1) by molecular biological and immunohistochemical methods. Histological examination of the brains revealed variable degrees of lymphohistiocytic encephalitis or meningoencephalitis, characterised predominantly by perivascular mononuclear infiltrates. Two cases could be attributed to PCV-2 infection by in situ hybridisation: viral nucleic acid was found in the mesencephalon, the cerebellum and the medulla oblongata, mainly in the cytoplasm of macrophages, endothelial cells and some glial cells, which were predominantly found in the meninges and around blood vessels. Real-time PCR detected PCV-2 DNA in brain samples from seven other pigs. There was no evidence of PRRSV, BDV, SuHV-1, PEVs or OvHV-2 in any of the brain samples examined.

Minipigs have become popular pets in recent years. Therefore, an increasing number of veterinarians are being challenged by specific problems of these animals. This retrospective study gives an overview on the diagnoses and therapeutic interventions of the patients submitted to the clinic for swine at the University of Veterinary Medicine Vienna during the last 6 years (n=48). Most frequently, colic symptoms of the gastro-intestinal tract (n=12) and orthopaedic locomotion disorders (n=10), mainly due to accidents or long claws, could be observed, followed by urogenital tract and skin disorders (n=4 each). Therapeutic interventions are discussed with regard to medical aspects as well as statutory provisions.