Newcastle disease virus: Detection and characterization by PCR of recent German isolates differing in pathogenicity.

The fusion (F) protein plays an important role in determining the virulence of Newcastle disease virus (NDV) strains. A reverse transcriptase-polymerase chain reaction (RT-PCR) is described which amplifies a 362 bp fragment encompassing the region of the F protein most important for pathogenicity. A specific PCR product was obtained independent of strain, pathogenicity and host of origin.

A simple statistical approach for assessing inter-laboratory comparison tests for classical swine fever diagnosis.

The aim of this study was to compare on an objective basis the results obtained during five classical swine fever (CSF) ring tests conducted in Germany between 1999 and 2003. A novel and simple statistical approach used in behavioural sciences was used. For each ring test, the regional laboratories received a panel of five lyophilized pig sera.

Diagnostic evaluation of a real-time RT-PCR assay for routine diagnosis of classical swine fever in wild boar.

The aim of the study was to evaluate a real-time reverse transcription-polymerase chain reaction (RT-PCR) assay for routine diagnosis of classical swine fever (CSF) in wild boar by using 1000 spleen homogenates that were tested previously negative by virus isolation and 26 homogenates from which CSF virus could be isolated. All 26 positive samples in the virus isolation assay also were found to be positive in real-time RT-PCR. Additionally further 10 samples were detected by real-time RT-PCR out of the 1000 negative samples in the virus isolation.

Characterization of avian paramyxovirus type 1 strains isolated in Germany during 1992 to 1996.

In Germany all avian paramyxoviruses (APMV) isolated in regional laboratories are collected and characterized by the National Reference Laboratory. From 1992 until 1996, 635 APMV-1 virus isolates were submitted from almost all regions. Of these viruses, 371 were isolated from chickens, 39 from other poultry, 171 from pigeons and 54 from exotic birds.

Engineering glycoprotein B of bovine herpesvirus 1 to function as transporter for secreted proteins: a new protein expression approach.

Glycoprotein B (gB) of bovine herpesvirus 1 (BHV-1) is essential for BHV-1 replication and is required for membrane fusion processes leading to virus penetration into the target cell and direct spreading of BHV-1 from infected to adjacent noninfected cells. Like many of the herpesvirus gB homologs, BHV-1 gB is proteolytically processed by furin, an endoproteinase localized in the trans-Golgi network. Cleavage by furin is a common mechanism for the activation of a number of viral fusion (F) proteins.

Prevention of virus persistence and protection against immunopathology after Borna disease virus infection of the brain by a novel Orf virus recombinant.

The Parapoxvirus Orf virus represents a promising candidate for novel vector vaccines due to its immune modulating properties even in nonpermissive hosts such as mouse or rat. The highly attenuated Orf virus strain D1701 was used to generate a recombinant virus (D1701-VrVp40) expressing nucleoprotein p40 of Borna disease virus, which represents a major antigen for the induction of a Borna disease virus-specific humoral and cellular immune response. Infection with Borna disease virus leads to distinct neurological symptoms mediated by the invasion of activated specific CD8+ T cells into the infected brain.

Interferon-gamma response of PBMC indicates productive pseudorabies virus (PRV) infection in swine.

In Chinese Meishan/German Landrace cross-bred swine F2 generation interferon gamma (IFN-gamma) production by peripheral blood mononuclear cells (PBMC) was determined directly ex vivo at different time points after survival of a virulent pseudorabies virus (PRV) infection. This reactivity was compared with the reactivity of naïve PBMC. Significant IFN-gamma production was determined in ELISA and ELISPOT only after in vitro PBMC re-stimulation with PRV and not with the closely related bovine herpesvirus BHV-1.

Potential risk factors for bovine Neospora caninum infection in Germany are not under the control of the farmers.

In the German state of Rhineland-Palatinate, herds were identified that were likely to have a Neospora caninum sero-prevalence > or = 10% by using a bulk milk ELISA. Individual herd data were obtained by a questionnaire. Univariate logistic regression showed that bulk milk positive farms had a significantly higher chance to report an increased abortion rate than negative farms (P(Wald)<0.

The ontogeny of MHC class I expression in rainbow trout (Oncorhynchus mykiss).

In the present study, clonal rainbow trout (Oncorhynchus mykiss) embryos and larvae were assayed for the expression of key molecules involved in specific cell-mediated cytotoxicity using an anti-MHC class I monoclonal Ab and by RT-PCR using specific primers derived from classical MHC class I (class Ia), TCR and CD8. Whereas RT-PCR revealed that MHC class Ia and CD8 were expressed from at least 1 week after fertilisation (p.f.

Glycosylation deficiency at either one of the two glycan attachment sites of cellular prion protein preserves susceptibility to bovine spongiform encephalopathy and scrapie infections.

The conversion into abnormally folded prion protein (PrP) plays a key role in prion diseases. PrP(C) carries two N-linked glycan chains at amino acid residues 180 and 196 (mouse). Previous in vitro data indicated that the conversion process may not require glycosylation of PrP.

Establishment of a new bovine leukosis virus producing cell line.

Due to the prevalence of different bovine leukosis virus (BLV) species in the cattle population in Europe, problems may arise in the serological diagnosis of BLV infections. In addition, earlier investigations demonstrated that contamination of the BLV antigen-producing cell culture systems by bovine viral diarrhea virus (BVDV) may give rise to misinterpretation of serological test results after BVDV vaccination of cattle. By co-cultivation of peripheral leukocytes of a BLV-infected cow with a permanent sheep kidney cell line, a new BLV-producing cell line named PO714 was established.

Development of maternal antibodies after oral vaccination of young female wild boar against classical swine fever.

An experimental study was performed to investigate the development of maternal antibodies after oral immunisation of young female wild boar against classical swine fever (CSF) using C-strain vaccine. Our results demonstrated that maternal antibodies do not persist in the offspring for more than 3 months. Based on the neutralising serum antibody titres, we assume that piglets of wild sows vaccinated orally twice or immunised once a long time before conception have protective antibodies for approximately 2 months.

Studies on the virulence of two field isolates of the classical Swine Fever virus genotype 2.3 rostock in wild boars of different age groups.

The virulence of two isolates of the classical swine fever virus (CSFV) was studied in experimentally infected wild boars of different ages. The isolates, originating from wild boars shot in Mecklenburg-Western Pomerania (isolate '1829-NVP') and in Rhineland-Palatinate (isolate '11722-WIL'), belong to the genetic subgroup 2.3 Rostock.

Neuronal accumulation of abnormal prion protein in sheep carrying a scrapie-resistant genotype (PrPARR/ARR).

The susceptibility of sheep to scrapie infection is influenced by prion gene alleles, which are modulated by polymorphic variations corresponding to amino acid positions 136, 154 and 173 of the prion protein (PrP). As no unquestioned report of a diseased sheep carrying homozygous alleles encoding alanine, arginine and arginine (PrPARR) at these sites has been published to date, sheep of this genotype are believed to be scrapie resistant. After the introduction of large-scale rapid testing for scrapie, a number of so-called 'atypical' scrapie cases have been found in Germany and elsewhere.

A novel protein expression strategy using recombinant bovine respiratory syncytial virus (BRSV): modifications of the peptide sequence between the two furin cleavage sites of the BRSV fusion protein yield secreted proteins, but affect processing and function of the BRSV fusion protein.

The bovine respiratory syncytial virus (BRSV) fusion (F) protein is cleaved at two furin cleavage sites, which results in generation of the disulfide-linked F(1) and F(2) subunits and release of an intervening peptide of 27 aa (pep27). A series of mutated open reading frames encoding F proteins that lacked the entire pep27, that contained an arbitrarily chosen 23 aa sequence instead of pep27 or in which pep27 was replaced by the amino acid sequences for the bovine cytokines interleukin 2 (boIL2), interleukin 4 (boIL4) or gamma interferon (boIFN-gamma) was constructed. Transient expression experiments revealed that the sequence of the intervening peptide influenced intracellular transport, maturation of the F protein and F-mediated syncytium formation.

B cell and macrophage response in chicks after oral administration of Salmonella typhimurium strains.

Despite the fact that, in a number of countries, vaccination programmes are extensively used to control Salmonella infection in poultry, information on the immune mechanisms, especially the cellular response, is still needed. The aim of the study was to characterise the B cell and macrophage response in caecum (IgA+, IgM+, IgG+ cells, macrophages), bursa of Fabricius (IgM+ cells, macrophages), and spleen (IgM+ cells) of chicks after oral administration of a non-attenuated Salmonella (S.) typhimurium wild-type strain (infection) or an attenuated commercial live S.

Correlation between invasion of Caco-2 eukaryotic cells and colonization ability in the chick gut in Campylobacter jejuni.

In an in vitro cell culture model using Caco-2 cells the adhesion and invasion properties of 11 Campylobacter (C.) jejuni isolates of different origin were studied. Additionally, we investigated the colonization ability of the strains in a chick model.

Recent isolates of parapoxvirus of Finnish reindeer (Rangifer tarandus tarandus) are closely related to bovine pseudocowpox virus.

Cases of papular stomatitis in Finnish reindeer have been reported for many years. The causative agent was thought to be Orf virus (ORFV), one of the Parapoxviridae, although this assumption was based mainly on clinical symptoms, pathology and electron microscopy. Here sequence analyses of the viral DNA isolated from a recent outbreak of disease in 1999-2000 are presented in comparison to that isolated from earlier outbreaks in 1992-1994.

Molecular fingerprinting of Mycobacterium bovis subsp. caprae isolates from central Europe.

To study the dissemination of Mycobacterium bovis subsp. caprae, 79 European isolates from cattle, humans, and other hosts were examined by spoligotyping and IS6110 restriction fragment length polymorphism (RFLP) analysis. Among a total of 11 different spoligotypes identified, type C1 proved to be predominant (n = 62).

Spill-over of European bat lyssavirus type 1 into a stone marten (Martes foina) in Germany.

European bat lyssavirus type 1 (EBLV-1, genotype 5) is known to endemically circulate in insectivorous bat populations in Germany. In August 2001, a rabies suspect stone marten (Martes foina) was found in the city of Burg (Saxony-Anhalt, Germany) and was sent to the regional veterinary laboratory for routine rabies diagnosis. Whereas brain samples repeatedly tested negative in the fluorescent antibody test for classical rabies virus (genotype 1), the mouse inoculation test and the rabies tissue culture inoculation test yielded positive results.

Atypical scrapie cases in Germany and France are identified by discrepant reaction patterns in BSE rapid tests.

The intensified surveillance of scrapie in small ruminants in the European Union (EU) has resulted in a substantial increase of the number of diagnosed cases. Four rapid tests which have passed the EU evaluation for BSE testing of cattle are also recommended currently and used for the testing of small ruminants by the EU authorities. These tests include an indirect ELISA (cELISA), a colorimetric sandwich ELISA (sELISA I), a chemiluminescent sandwich ELISA (sELISA II), and a Western blot (WB).

Comparison and standardisation of serological methods for the diagnosis of Neospora caninum infection in bovines.

Various existing serological tests were compared with a standard panel of 523 sera in a multicentred study across Europe. Well characterised sera from animals that were experimentally or naturally infected with Neospora caninum as well as sera from cattle deemed uninfected with N. caninum were provided by the participants of the study and analysed in several commercial (CHEKIT Dr.

Simultaneous deletion of pseudorabies virus tegument protein UL11 and glycoprotein M severely impairs secondary envelopment.

The pseudorabies virus (PrV) proteins UL11, glycoprotein E (gE), and gM are involved in secondary envelopment of tegumented nucleocapsids in the cytoplasm. To assess the relative contributions of these proteins to the envelopment process, virus mutants with deletions of either UL11, gM, or gE as well as two newly constructed mutant viruses with simultaneous deletions of UL11 and gE or of UL11 and gM were analyzed in cell culture for their growth phenotype. We show here that simultaneous deletion of UL11 and gE reduced plaque size in an additive manner over the reduction observed by deletion of only UL11 or gE.

Influence of tegument proteins of pseudorabies virus on neuroinvasion and transneuronal spread in the nervous system of adult mice after intranasal inoculation.

Pseudorabies virus (PrV) is a neurotropic alphaherpesvirus that, after intranasal infection of adult mice, enters peripheral neurons and propagates to the central nervous system. In recent years we have analyzed the contribution of virus-encoded glycoproteins to neuroinvasion and transneuronal spread (reviewed in T. C.